Compositions for prevention of adhesions and other barrier applications

The present invention is generally in the field of tissue application formulations for the prevention of adhesions and other barrier applications.

BACKGROUND OF THE INVENTION [0002] Adhesions may be present from birth (congenital) or may form after surgery in the abdominal cavity or after inflammation. Most adhesions are usually formed after surgery. The adhesions occur more often after procedures in the colon, appendix or uterus than after surgery within other organs, such as the stomach, gall bladder or pancreas. The risk of adhesions increases over time after surgery.

[0003] Adhesions in the abdominal cavity are strands of fibrous scar tissue that form on the organs in the abdominal cavity, causing the organs to adhere to each other or to the abdominal wall. Intestinal adhesions are strands of fibrous tissue that connect the intestinal loops with each other; intestines with other abdominal organs; or intestines with the abdominal wall. These bands can pull intestinal sections out of their position and can block the passage of food. In people living in developed countries, this scar tissue develops most often after surgical operations in the abdominal cavity, during which the surgical team deals with organs and temporarily shifts them from their correct positions. Scar tissue can also develop in people who have developed peritonitis, an infection that has spread to the membrane, which covers the abdominal organs. Peritonitis usually occurs after appendicitis or other abdominal infections. Another cause of adhesions is endometriosis, an inflammation that affects some women and can affect the abdominal cavity and a serious abdominal injury, including cesarean cuts. [0004] Adhesions are the main cause of intestinal obstruction. Adhesions can cause partial or complete intestinal obstruction. Symptoms resulting from adhesions depend on the degree and location of the obstruction. Symptoms include cramps, abdominal pain, vomiting, bloating, inability to flow gas, and constipation. However, in a small number of people who have adhesions, the fibrous bands of scar tissue block the intestines completely or partially. This blockage is called intestinal obstruction, and about 5% of cases lead to death. Sometimes the intestinal area, which is affected by adhesions, can be blocked and then unblocked, causing symptoms that appear and disappear. In about 10% of cases of small intestinal obstruction, part of the intestine twists strongly around the band of adhesions. It cuts off the proper supply of blood to the twisted intestine, which is a disorder known as the disorder, causing the part of the intestine to die. When this happens, the person must immediately undergo a surgical procedure. The mortality rate is as high as 37% in people who experience the implantation. which is a disorder known as a disorder that causes part of the intestine to die. When this happens, the person must immediately undergo a surgical procedure. The mortality rate is as high as 37% in people who experience the implantation. which is a disorder known as a disorder that causes part of the intestine to die. When this happens, the person must immediately undergo a surgical procedure. The mortality rate is as high as 37% in people who experience the implantation.

[0005] Percutaneous epidural adhesion and spinal endoscopic adhesion are invasive pain management techniques used to treat patients with persistent low back pain due to epidural scarring. Standard epidural steroid injections are often ineffective, especially in patients with previous back surgery. Adhesions in the epidural space can prevent the drug from flowing to the target area; lysis of these adhesions may improve the delivery of the drug to the affected areas, which may improve the therapeutic efficacy of the injected drugs. Preventing such adhesions would, however, be more beneficial.

[0006] Many different materials have been tried as adhesion prevention agents. Most of them are hydrogels, which are used as solutions during surgery. The effectiveness of these materials differs due to rapid degradation and / or the inability to form a sufficiently thick barrier. Other materials only work in combination with anti-proliferative drugs. None of these materials have been found to be effective in a high fluidity environment that normally occurs during surgery due to the condensation and leakage of other body fluids.

[0007] In US Patent Nos. 5,670,483; 6548630, and 7098028 by Zhang et al. amphiphilic peptides having alternating hydrophobic and hydrophilic residues are described. Zhang argues that membranes are potentially useful in biomaterial applications, such as slow-spreading drug delivery systems, artificial skin and separating matrices, and as experimental models in Alzheimer's disease and scrapie infection. However,

Zhang does not disclose the use of such materials to prevent adhesions.

[0008] WO 2007/142757 and USSN 11/411745 describe compositions comprising peptides with alternating hydrophilic and hydrophobic monomers that allow them to self-assemble under physiological conditions that are formulated for application to wounds.

[0009] WO 2006/116524 relates to compositions comprising nanoscale structured materials or their precursors, which may contain other substances, such as vasoconstrictors.

[0010] WO 2006/014570 relates to amphiphilic peptide chains alternating hydrophilic and hydrophobic amino acids, said peptide comprising at least amino acids that are complementary and structurally compatible and self-assembling into a macroscopic scaffold of a beta sheet, wherein in the peptide at least about 75 % of chains have the same sequence.

[0011] WO 2004/007532 relates to materials comprising webs, filaments or fibers characterized in that each of the ribbons, filaments or fibers has an antiparallel arrangement of peptides in the tape-like sub-structure of the beta-sheet.

[0012] However, these applications do not describe the use of such materials to prevent adhesions.

[0013] The object of the present invention is therefore to provide a composition for preventing or minimizing adhesions and for other barrier applications that can be applied to tissues or cells that bleed or in the presence of fluids.

[0014] Another object of the present invention is to provide a composition that can be formulated as a bandage, spray, coating or powder.

[0015] It is yet another object of the present invention to provide a composition that can be used to prevent adhesions, but which is sufficiently clear to allow the physician to view and work through the material.

BRIEF SUMMARY OF THE INVENTION [0016] The invention is as defined in the appended claims.

[0017] Compositions comprising peptides that self-assemble under physiological conditions are formulated onto tissues to prevent adhesions or other barrier applications, such as minimizing contamination or infection (eg from bacteria, fungi, viruses or other pathogenic agents), limiting the spread of metastasis after tumor surgery, or to provide a therapeutic, diagnostic or prophylactic agent in a confined region after substantial bleeding or fluid leakage.

[0018] In one embodiment, the composition comprises self-assembling peptides with a length in the range of 6 to 200 amino acids and they include a sequence of amino acid residues corresponding to one or more of the following formulas I-IV:

((Xaa^neu-Xaa⁺) X (Xaa^neu-Xaa^-) y) n (I) ((Xaa^neu-Xaa^-) X (Xaa^neu-Xaa⁺) y) n (II) ((Xaa⁺-Xaa^neu) X (Xaa^--Xaa^neu) y) n (III) ((Xaa^--Xaa^neu) X (Xaa⁺-Xaa^neu) y) n (IV) wherein Xaa^neu means an amino acid residue having an inert charge; Xaa⁺ is an amino acid residue having a positive charge; Xaa^- is an amino acid residue having a negative charge; x and y are independently integers with a value of 1, 2, 3 or 4; and n is an integer having a value of 1-5, wherein the self-assembling peptides can form a self-assembling barrier structure that inhibits or prevents the passage of body fluid or intracellular substance through the structure, the self-assembling peptides further comprising an amino acid sequence, which interacts with the extracellular matrix, wherein the amino acid sequence anchors the self-assembling peptides in the extracellular matrix, for use in a method of preventing adhesions after surgery or injury at a demanding site.

[0020] The concentration of self-assembling peptides in any given formulation may vary and may range from about 0.1% to 99%, inclusive, preferably between 0.1% and 10%. In one embodiment, the concentration of self-assembling peptides (e.g., in a liquid formulation) may be about 0.1-3.0% (1-30 mg / mL) (e.g., 0.1-1.0%, 1.0-2 , 0%, 2.0-3.0% or 1.0-3.0%). The concentration of self-assembling peptides may be higher in the main solutions and in solid preparations (e.g. powdered). The concentration of self-assembling peptides in solid preparations may approach 100% (e.g., the concentration of self-assembling peptides in the composition may be 95, 96, 97, 98, 99% or more (e.g., 99.99%)). Regardless of whether they are in liquid or solid form, the peptides can be brought to the desired concentration before use by adding a pharmaceutically acceptable diluent (e.g., deionized water), fillers or oil. The formulations may contain a pharmaceutically acceptable carrier or therapeutic, prophylactic or diagnostic agents. These include, but are not limited to, anti-inflammatory agents, vasoactive agents, anti-infective agents, anesthetics, growth factors and / or cells. Metals can be added as chelators or to further reduce adhesions.

[0021] The formulation may be administered, as appropriate, for the treatment of one or more disorders or conditions, such as those listed above. For example, the preparation can be used after repairing wounds or during lung surgery, eyes or dura mater, or after epidural or lumbar puncture, in order to prevent or minimize the formation of adhesions. Self-assembling peptides can also be used to prevent adhesions after laminectomy (e.g., equine tail adhesions induced by laminectomy) and adhesions after decompression of the core (e.g., adhesions of the meninges). Self-assembling peptides can also be used as a block or filter of the immune system, especially in the case of wounding an organ or tissue, to prevent the accumulation of red blood cells and / or platelet aggregation at the wound site.

[0022] In some embodiments, self-assembling peptides may allow the passage of selected materials, preventing the passage or introduction of other materials such as bacteria, viruses, fungi, etc.

[0023] The formulation may be used as a hydrogel, a layered preparation, including oil or as a spray. In one embodiment, the formulation is provided as a dry or lyophilized powder that can be administered directly in the form of a powder or tablet, disc or wafer that hydrates at the site of administration, or it can be suspended or dissolved in a liquid and most preferably in water, and administered as a spray, a brush form, an injection or a hydrogel containing a material such as chitin, collagen, alginate or a synthetic polymer. In a preferred embodiment, self-assembling peptides are provided in combination with an oil, the combination forming a layered preparation. In yet another embodiment, the formulation is provided in the form of a bandage, foam or matrix in which the self-assembling peptides can be dispersed or absorbed. The formulation may also be in the form of sutures, tape or glue. The liquid formulations may be provided in a syringe or pipette having a cylinder containing a composition comprising self-assembling peptides and means for ejecting the composition from the open end of the syringe or pipette (e.g., plunger or pear). The syringe may consist of one or more compartments such that, during use, the self-assembling peptides are mixed with one or more other agents. The compartments may also contain excipients, such as a hydrogel-forming material or glue in one compartment, and self-assembling peptides in another compartment. In another embodiment, one compartment may comprise self-assembling peptides in lyophilized form or molecules, and another compartment may include a solution for dissolving or hydration of peptides, or mixed with other application powders in dry form. The liquid and powdered compositions are stable, preferably over a period of more than one year, more preferably above two years, and most preferably above three years.

[0024] One or more of the compositions described herein may be assembled in kits containing instructions for use. For example, the kits may include a bio-compatible composition comprising self-assembling peptides (or a concentrated solution thereof or a powdered formulation with a diluent) and a vasoconstrictor, a colorant or painkiller, or anesthetic and instructions for combining them (if they have not already been combined) and use (e.g., dilution and administration). The kits may further comprise one or more additional measures described herein. These agents may be present in a self-assembling composition or may be packaged separately and may include one or more of a type of biological cell, antibiotic or other therapeutic agent, collagen, anti-inflammatory agent, growth factor or nutrient. The kit may also contain one or more syringes (e.g., a cylindrical syringe or a medical syringe), a needle, a pipette, gauze, sponge, cotton swab, swabs, bandage, disinfectant, surgical sutures, scissors, scalpel, sterile liquid, a container for sprays, including those in which the liquid solution is sprayed by a simple hand pump, a sterile container or disposable gloves. The kit may also contain one or more additives to change the kinetics of the material assembly depending on the environment in which the self-assembling peptides are to be used. swabs, bandage, disinfectant, surgical sutures, scissors, scalpel, sterile liquid, spray tank, including those in which the liquid solution is sprayed by a simple hand pump, a sterile container or disposable gloves. The kit may also contain one or more additives to change the kinetics of the material assembly depending on the environment in which the self-assembling peptides are to be used. swabs, bandage, disinfectant, surgical sutures, scissors, scalpel, sterile liquid, spray tank, including those in which the liquid solution is sprayed by a simple hand pump, a sterile container or disposable gloves. The kit may also contain one or more additives to change the kinetics of the material assembly depending on the environment in which the self-assembling peptides are to be used.

DETAILED DESCRIPTION OF THE INVENTION

I. Formulations [0025] "Adherents" as used herein generally refer to fibrous tissue and / or scar tissue associated with the surface of an organ and / or tissue capable of combining, covering or distorting organs and / or tissue. Adhesions can be caused by previous infection and / or surgery. Adhesions may occur in various areas of the body, including, but not limited to, the pelvic region, abdominal cavity, intestines and reproductive organs such as fallopian tubes or ovaries.

[0026] "Biocompatible", as used herein, refers to compatibility with a living tissue or living system, not being toxic, harmful or physiologically reactive, and without causing immunological rejection.

[0027] "Complementary" means having the ability to form interactions in the form of ionic or hydrogen bond between hydrophilic moieties of adjacent peptides in the structure. Each hydrophilic residue in the peptide binds via hydrogen or ionic bonds to the hydrophilic residue at the adjacent peptide or is exposed to the solvent. The bond may also include van der Waals forces.

[0028] An "effective amount", in reference to an active agent, such as a self-assembling peptide or biomolecule, pharmaceutical agent, etc., refers to the amount necessary to elicit the desired biological response. Those of ordinary skill in the art will appreciate that an effective amount of the agent may vary depending on such factors as the desired biological endpoint, the agent to be delivered, the nature of the site to which the agent is delivered, the nature of the conditions for which it is administered. measure, etc. For example, an effective amount of a composition for treating diabetic retinopathy may be an amount sufficient to promote recovery to a greater extent than would be the case in the absence of a composition.

[0029] "Hemostasis" refers to the cessation of bleeding.

[0030] "Prevention" refers to preventing a condition, condition or disease from occurring, or a symptom or symptom thereof, or an increase in the severity of such symptoms. Prevention includes reducing the risk of a condition, condition or disease, or their symptom or symptom or severity.

[0031] "Repair", as used in connection with tissue repair in various embodiments of the invention, may include any aspect of the anatomical or functional restoration of tissue prior to injury, deterioration or other damage. For example, this may include restoring physical continuity between tissue parts that have been separated by injury, deterioration or other damage. Preferably, such restoration of the physical continuity involves repositioning or re-joining part of the tissue without appreciably separating it by the type of tissue that was not present prior to injury, e.g. scar tissue. Repair may or may not include the growth or development of new tissue. "Repair" and "healing" are used interchangeably here.

[0032] "Self-assembly", as used herein, refers to the assembly of molecules into specific, non-covalently associated assemblies that are held together by intermolecular forces. Folding can be spontaneous or induced.

II. Self-assembling materials

A. Self-assembling peptides [0033] The term "peptide" as used herein includes "polypeptide", "oligopeptide" and "protein" and refers to a chain of at least two a-amino acid residues joined together by covalent linkages (e.g., peptide bonds). Useful peptides may vary in length, provided that they retain their self-dispensing capability in a range useful for one or more of the purposes described herein. The number of amino acid residues in the peptide may range from about 6 to about 200 residues, preferably from about 6 to about 64 residues, more preferably from about 8 to about 36 residues, and most preferably from about 8 to about 24 residues. The peptides may be at least six amino acids long (e.g., eight or 10 amino acids), at least 12 amino acids (e.g., 12 or 14 amino acids) or at least 16 amino acids (e.g., 16, 18, 20, 22 or 24 amino acids). Peptides that are less than 100 amino acid residues, more preferably less than about 50 amino acids, can be self assembled more easily. In one embodiment, the peptide has from about 8 to about 16 residues. In another embodiment, the peptide has from about 12 to about 20 residues. In yet another embodiment, the peptide has from about 16 to about residues. A "peptide" may refer to a single peptide or to a collection of peptides having the same or different sequences, each of which may contain naturally occurring α-amino acid residues, non-naturally occurring α-amino acid residues, and combinations thereof. Α-amino acid analogs are also known in the art and may be used alternatively. In particular, Do-amino residues may be used. more preferably less than about 50 amino acids, may be more easily self-deposited. In one embodiment, the peptide has from about 8 to about 16 residues. In another embodiment, the peptide has from about 12 to about 20 residues. In yet another embodiment, the peptide has from about 16 to about residues. A "peptide" may refer to a single peptide or to a collection of peptides having the same or different sequences, each of which may contain naturally occurring α-amino acid residues, non-naturally occurring α-amino acid residues, and combinations thereof. Α-amino acid analogs are also known in the art and may be used alternatively. In particular, Do-amino residues may be used. more preferably less than about 50 amino acids, may be more easily self-deposited. In one embodiment, the peptide has from about 8 to about 16 residues. In another embodiment, the peptide has from about 12 to about 20 residues. In yet another embodiment, the peptide has from about 16 to about residues. A "peptide" may refer to a single peptide or to a collection of peptides having the same or different sequences, each of which may contain naturally occurring α-amino acid residues, non-naturally occurring α-amino acid residues, and combinations thereof. Α-amino acid analogs are also known in the art and may be used alternatively. In particular, Do-amino residues may be used. In yet another embodiment, the peptide has from about 16 to about residues. A "peptide" may refer to a single peptide or to a collection of peptides having the same or different sequences, each of which may contain naturally occurring α-amino acid residues, non-naturally occurring α-amino acid residues, and combinations thereof. Α-amino acid analogs are also known in the art and may be used alternatively. In particular, Do-amino residues may be used. In yet another embodiment, the peptide has from about 16 to about residues. A "peptide" may refer to a single peptide or to a collection of peptides having the same or different sequences, each of which may contain naturally occurring α-amino acid residues, non-naturally occurring α-amino acid residues, and combinations thereof. Α-amino acid analogs are also known in the art and may be used alternatively. In particular, Do-amino residues may be used.

[0034] Furthermore, one or more amino acid residues in the self-assembling peptide can be altered or derivatized by the addition of one or more chemical units including, but not limited to, acyl groups, carbohydrate groups, carbohydrate chains, phosphate groups, farnesyl groups, groups isofarnesyl, fatty acid groups or a linker that allows coupling or functionalization of the peptide. For example, one or both ends of a given peptide may be modified. For example, the carboxyl and / or amino groups of the carboxyl and amino termination residues can be suitably protected or unprotected. The load at the end can also be modified. For example, a group or radical, such as an acyl group (RCO-, wherein R is an organic group (e.g. an acetyl group (CH3CO-)) may be present at the N-terminus of the peptide to neutralize the "additional" positive charge that may otherwise be present (e.g., a non-side load of the N-terminal amino acid). Similarly, a group such as an amino group (RNH- where R is an organic group (e.g., an amino group -NH2)) can be used to neutralize the "additional" negative charge that may otherwise be present at the C-terminus (e.g. non-side chain of the C-terminal amino acid residue). Where an amine is used, the C-terminus contains an amide moiety (-CONHR). The neutralization of loads at the end can facilitate self-assembly. An ordinary specialist in the field will be able to select other suitable groups. load not derived from the side chain of the N-terminal amino acid). Similarly, a group such as an amino group (RNH- where R is an organic group (e.g., an amino group -NH2)) can be used to neutralize the "additional" negative charge that may otherwise be present at the C-terminus (e.g. non-side chain of the C-terminal amino acid residue). Where an amine is used, the C-terminus contains an amide moiety (-CONHR). The neutralization of loads at the end can facilitate self-assembly. An ordinary specialist in the field will be able to select other suitable groups. load not derived from the side chain of the N-terminal amino acid). Similarly, a group such as an amino group (RNH- where R is an organic group (e.g., an amino group -NH2)) can be used to neutralize the "additional" negative charge that might otherwise be present at the C-terminus (e.g. non-side chain of the C-terminal amino acid residue). Where an amine is used, the C-terminus contains an amide moiety (-CONHR). The neutralization of loads at the end can facilitate self-assembly. An ordinary specialist in the field will be able to select other suitable groups. which may otherwise be present at the C-terminus (e.g., a non-side-load of the C-terminal amino acid residue). Where an amine is used, the C-terminus contains an amide moiety (-CONHR). The neutralization of loads at the end can facilitate self-assembly. An ordinary specialist in the field will be able to select other suitable groups. which may otherwise be present at the C-terminus (e.g., a non-side-load of the C-terminal amino acid residue). Where an amine is used, the C-terminus contains an amide moiety (-CONHR). The neutralization of loads at the end can facilitate self-assembly. An ordinary specialist in the field will be able to select other suitable groups.

[0035] Suitable peptides may also be branched, in which case they will contain at least two amino acid polymers, each of which consists of at least three amino acid residues joined by peptide bonds. The two amino acid polymers may be joined by a bond other than the peptide bond.

[0036] While peptide sequences may vary, useful sequences include those that transfer the amphiphilic character to peptides (e.g., peptides may contain approximately the same number of hydrophobic and hydrophilic amino acid residues), and peptides may be complementary and structurally compatible. Complementary peptides have the ability to form ionic or hydrogen bonds between residues (e.g., hydrophilic residues) in neighboring peptides in the structure. For example, one or more hydrophilic residues in a peptide may form a hydrogen or ionic bond with one or more hydrophilic residues in an adjacent peptide. The hydrophilic residues are those residues that typically contain a polar functional group or a functional group that carries the charge under physiological conditions. Exemplary functional groups include, but not exclusively, carboxylic acid groups, amino groups, sulphate groups, hydroxyl groups, halo groups, nitro groups, phosphate groups, etc. Hydrophobic residues are those moieties that contain non-polar functional groups. Exemplary functional groups include, but are not limited to, alkyl groups, alkene groups, alkyne groups, and phenyl groups.

[0037] In one embodiment, the hydrophilic moiety has the formula -NH-CH (X) -COO-, wherein X has the formula (CH2) yZ, wherein y = 0-8, preferably 1-6, more preferably 1-4, and most preferably 1-3 and Z is a polar or charged functional group including, but not limited to, a carboxylic acid group, an amino group, a sulfate group, a hydroxyl group, a halo group, a nitro group, a phosphate group, or a quaternary amine functional group.

The alkyl chain may be in a linear, branched or cyclic system. X may also contain one or more heteroatoms in the alkyl chain and / or X may be substituted with one or more additional substituents. In a preferred embodiment, Z is a carboxylic acid group or an amino group. In one embodiment, the hydrophobic moiety has the formula -NH-CH (X) -COO-, wherein X has the formula (CH2) yZ, where y = 0-8, preferably 1-6, more preferably 1-4, and more preferably 1-3. and Z is a non-polar functional group including, but not limited to, an alkyl group, an alkene group, an alkyne group, or a phenyl group. The alkyl, alkene or alkyne chain may be in a linear, branched or cyclic system. X may also contain one or more heteroatoms in the alkyl chain and / or X may be substituted with one or more additional substituents. In a preferred embodiment, X is an alkyl group, such as a methyl group.

[0038] Where self-assembling peptides are used, it is believed that their side chains (or R groups) divide into two surfaces, a polar surface with positive and / or negatively charged ionic side chains (e.g., side chains containing groups - OH, -NH, -CO 2 H, or -SH) and a non-polar side with side chains that are considered inert or uncharged at physiological pH (e.g., the side chain of alanine residues or residues having other hydrophobic groups). The positively charged and negatively charged amino acid residues on the polar surface of one peptide can form complementary ion pairs with oppositely charged residues of another peptide. These peptides can thus be referred to as ionic, self-complement peptides. If the ionic residues are alternately one positively charged and one negatively charged residue on the polar surface (- + - + - + - +), the peptides can be described as "module I"; if the ionic residues are alternately two positively and two negatively charged residues (- ++ - ++) on the polar surface, the peptides are described as "module II"; if the ionic residues are alternately three positive and three negative residues (+++ --- +++ ---) on the polar surface, the peptides are described as "module III"; if the ionic residues are alternately four positive and four negatively charged residues (++++ ---- ++++ ----) on the polar surface, they are described as "module IV". A peptide having four repeating EAKA sequence units (SEQ ID NO: 111) can be designated EAKA16-I (SEQ ID NO: 410), and peptides with different sequences can be described by the same convention.

[0039] Unpaired moieties may interact (e.g., form hydrogen bonds, etc.) with a solvent. Peptide-peptide interactions may also include van der Waals forces and / or forces that are not covalent bonds. The peptides are structurally compatible when they are able to maintain a sufficiently constant intrapeptide distance to allow self-assembly and structure formation. The intrapeptide distance may vary. An "intrapeptide distance", as used herein, refers to the average representative value of the distance between adjacent amino acid residues. In one embodiment, the intrapeptide distance is less than about 4 angstroms, preferably less than about 3, more preferably less than about 2 angstroms, and most preferably less than about 1 angstrom. The intrapeptide distance may, however, be greater.

[0040] The structures described herein may be formed by self-assembly of the peptides described in US Patent Nos. 5670483; 5955343; 6548630; and 6800481, Zhang et al .; Holmes et al., Proc. Natl. Acad. Sci. USA, 97: 6728-6733 (2000); Zhang et al., Proc. Natl. Acad. Sci. USA, 90: 3334-3338 (1993); Zhang et al., Biomaterials, 16: 1385-1393 (1995); Caplan et al., Biomaterials, 23: 219-227 (2002); Leon et al., J. Biomater. Sci. Polym. Editions, 9: 297-312 (1998); and Caplan et al., Biomacromolecules, 1: 627-631 (2000).

[0041] Self-assembling peptides comprising alternating hydrophobic and hydrophilic amino moieties can be used. Examples of representative hydrophobic and hydrophilic peptides are listed in Table 1.

Table 1. Representative self-assembling peptides

No. Sequence (NC)

T n-SGSGSGSGSGSGSGSG-c (SEQ ID NO: 2)

2. n-SASASASASASASASAS-c (SEQ ID NO: 3)

3. n-SVSVSVSVSVSVSVSV-c (SEQ ID NO: 4)

4. n-SLSLSLSLSLSLSLSL-c (SEQ ID NO: 5)

5. n-SISISISISISISI-c (SEQ ID NO: 6)

6. n-SMSMSMSMSMSMSMSM-c (SEQ ID NO: 7)

7. n-SFSFSFSFSFSFSFS-c (SEQ ID NO: 8)

8. n-SWSWSWSWSWSWSWSW-c (SEQ ID NO: 9)

9. n-SPSPSPSPSPSPSPS-c (SEQ ID NO: 10)

10. n-TGTGTGTGTGTGTGTG-c (SEQ ID NO: 11)

11. n-TATATATATATATATA-c (SEQ ID NO: 12)

12. n-TVTVTVTVTVTV-c (SEQ ID NO: 13)

13. n-TLTLTLTLTLTLTLTL-c (SEQ ID NO: 14)

14. n-TITITITITITITITI-c (SEQ ID NO: 15)

15. n-TMTMTMTMTMTMTMTM-c (SEQ ID NO: 16)

16. n-TFTFTFTFTFTFTFTF-c (SEQ ID NO: 17)

17. n-TWTWTWTWTWTWTWTW-c (SEQ ID NO: 18)

18. n-TPTPTPTPTPTPTPTP-c (SEQ ID NO: 19)

19. n-CGCGCGCGCGCGCGCG-c (SEQ ID NO: 20)

20. n-CACACACACACACACA-c (SEQ ID NO: 21)

21. n-CVCVCVCVCVCVCVCV-c (SEQ ID NO: 22)

22. n-CLCLCLCLCLCLCL-c (SEQ ID NO: 23)

23. n-CICICICICICICIC-c (SEQ ID NO: 24)

24. n-CMCMCMCMCMCMCMCM-c (SEQ ID NO: 25)

25. n-CFCFCFCFCFCFCFCFC-c (SEQ ID NO: 26)

26. n-CWCWCWCWCWCWCWC-c (SEQ ID NO: 27)

27. n-CPCPCPCPCPCPCPCP-c (SEQ ID NO: 28)

28. n-YGYGYGYGYGYGYGYG-c (SEQ ID NO: 29)

29. n-YAYAYAYAYAYAYAYAY-c (SEQ ID NO: 30)

30. n-YVYVYVYVYVYVYVYV-c (SEQ ID NO: 31)

31. n-YLYLYYYYYYYYYYYL-c (SEQ ID NO: 32)

32. n-YIYIYIYIYIYIYIYI-c (SEQ ID NO: 33)

33. n-YMmiMMayimM-c (SEQ ID NO: 34)

34. n-YFYFYFYFYFYFYFYF-c (SEQ ID NO: 35)

35. N-RESIDUAL-c (SEQ ID NO: 36)

36. n-YPYPYPYPYPYPYPYP-c (SEQ ID NO: 37)

37. n-NGNGNGNGNGNGNGNG-c (SEQ ID NO: 38)

38. n-NANANANANANANAN-c (SEQ ID NO: 39)

39. n-NVNVNVNVNVNVNVNV-c (SEQ ID NO: 40)

40. n-NLNLNLNLNLNLNLNL-c (SEQ ID NO: 41)

41. n-NINININININININI-c (SEQ ID NO: 42)

42. n-NMNMNMNMNMNMNMNM-c (SEQ ID NO: 43)

43. n-NFNFNFNFNFNFNFNF-c (SEQ ID NO: 44)

44. n-NWNWNWNWNWNWNWNW-c (SEQ ID NO: 45)

45. n-NPNPNPNPNPNPNPNP-c (SEQ ID NO: 46)

46. ​​n-QGQGQGQGQGQGQGQG-c (SEQ ID NO: 47)

47. n-QAQAQAQAQAQAQAQA-c (SEQ ID NO: 48)

48. n-QVQVQVQVQVQVQVQV-c (SEQ ID NO: 49)

49. n-QLQLQLQLQLQLQLQL-c (SEQ ID NO: 50)

50. n-QIQIQIQIQIQIQIQI-c (SEQ ID NO: 51)

51. n-QMQMQMQMQMQMQMQM-c (SEQ ID NO: 52)

52. n-QFQFQFQFQFQFQFQF-c (SEQ ID NO: 53)

53. n-QWQWQWQWQWQWQWQW-c (SEQ ID NO: 54)

54. n-QPQPQPQPQPQPQPQP-c (SEQ ID NO: 55)

55. n-AEAKAEAKAEAKAEAK-c (SEQ ID NO: 56)

56. n-RADARADARADARADA-c (SEQ ID NO: 1)

57. n-RAEARAEARAEARAEA-c (SEQ ID NO: 58)

58. n-OADAKADADADADADA-c (SEQ ID NO: 59) [0042] Other peptides or proteins may be used in combination with or alternatively with the disclosed self-assembling peptides or compositions. It should be noted that additional peptides may contain other self-assembling peptides or proteins. Alternatively, the peptide may be peptides that do not self-assemble. Representative additional peptides, proteins or chemically modified variants thereof include, but are not limited to, the peptides shown in Table 2.

Table 2. Additional peptides

1. Pmp-Y (Me) -ITNCP-Orn-Y-NH2 (SEQ ID NO: 60)

2. Mpr-YFQNCPR (SEQ ID NO: 61)

3. CYFQNCPRG-NH2 (SEQ ID NO: 62)

4. CYFQNCPR (SEQ ID NO: 63)

5. CY-Ile-QNCPRG-NH2 (SEQ ID NO: 64)

6. YFQN-Asu-PRG-NH2 (SEQ ID NO: 65)

7. Y-Ile-QN-Asu-PRG-NH2 (SEQ ID NO: 66)

8. Mpr-D-Pyridylanino-FQNCPRG-NH2 (SEQ ID NO: 67)

9. Deamino-Pen-YFVNCP-DR-G-NH2 (SEQ ID NO: 68)

10. Mpr-YFQNCPRG-NH2 (SEQ ID NO: 69)

11. Mpr-YFQNCP-DR-G-NH2 (SEQ ID NO: 70)

12. Mpr-YFQNCPK (SEQ ID NO: 71)

13. CYFQNCPKG-NH2 (SEQ ID NO: 72)

14. CYFQNCPK (SEQ ID NO: 73)

15. Mpr-YFVNCP-DR-G-NH2 (SEQ ID NO: 74)

16. CF-Ile-QNCP-Orn-G-NH2 (SEQ ID NO: 75)

17. Pmp-DY (OEt) -FVNCP-Cit-G-NH2 (SEQ ID NO: 76)

18. Pmp-Y (OEt) -FVNCPRG-NH2 (SEQ ID NO: 77)

19. Pmp-Y (Me) -FQNCPRG-NH2 (SEQ ID NO: 78)

20. Pmp-Y (Me) -IQNCP-Orn-G-NH2 (SEQ ID NO: 79)

21. G-DR-GDSP (SEQ ID NO: 80)

22. G-DR-GDSPASSK (SEQ ID NO: 81)

23. GPR

24. G-Pen-GRGDSPCA (SEQ ID NO: 82)

25. GRADSP (SEQ ID NO: 83)

26. GRGD-DS-P (SEQ ID NO: 84)

27. GRGDNP (SEQ ID NO: 85)

28. GRGDS (SEQ ID NO: 86)

29. GRGDSP (SEQ ID NO: 87)

30. GRGDSPC (SEQ ID NO: 88)

31. GRGDSPK (SEQ ID NO: 89)

32. GRGDTP (SEQ ID NO: 90)

33. GRGES (SEQ ID NO: 91)

<td>34.</td><td colspan="2">GRGESP (SEQ ID NO: 92)</td>

<td>35.</td><td>GRGETP (SEQ ID NO: 93)</td><td></td>

<td>36.</td><td>KGDS (SEQ ID NO: 94)</td><td></td>

<td>37.</td><td>GAVSTA (SEQ ID NO: 95)</td><td></td>

<td>38.</td><td>WTVPTA (SEQ ID NO: 96)</td><td></td>

<td>39.</td><td>TDVNGDGRHDL (SEQ ID NO: 97)</td><td></td>

<td>40.</td><td>REDV (SEQ ID NO: 98)</td><td></td>

<td>41.</td><td>RGDC (SEQ ID NO: 99)</td><td></td>

<td>42.</td><td>RGDS (SEQ ID NO: 100)</td><td></td>

<td>43.</td><td>RGDSPASSKP (SEQ ID NO: 101)</td><td></td>

<td>44.</td><td>RGDT (SEQ ID NO: 102)</td><td></td>

<td>45.</td><td>RGDV (SEQ ID NO: 103)</td><td></td>

<td>46.</td><td>RGES (SEQ ID NO: 104)</td><td></td>

<td>47.</td><td>SDGR (SEQ ID NO: 105)</td><td></td>

<td>48.</td><td>SDGRG (SEQ ID NO: 106)</td><td></td>

<td>49.</td><td>YRGDS (SEQ ID NO: 107)</td><td></td>

<td>50.</td><td>EGVNDNEEGFFSAR (SEQ ID NO: 108)</td><td></td>

<td>51.</td><td>YADSGEGDFLAEGGGVR (SEQ ID NO:</td><td>109)</td>

<td>52.</td><td>Glp-GVNDNEEGFFSARY (SEQ ID NO:</td><td>110)</td>

<td>pmp</td><td>= pyridoxamine phosphate</td><td></td>

<td>mpr</td><td>= 3-mercaptopropionyl</td><td></td>

<td colspan="2">Deamino-Pen = deaminopenicylamine</td><td></td>

<td>Pen</td><td>= penicillamine</td><td></td>

<td>Asu</td><td>= aminosuccinyl</td><td></td>

<td>OEt</td><td>= ethoxy group</td><td></td>

<td>Me =</td><td>methyl</td><td></td>

<td>Cit =</td><td>citrulline</td><td></td>

[0043] Other useful self-assembling peptides may be made, for example, which are different from those illustrated by a single amino acid residue or by a plurality of amino acid residues (e.g., by inclusion or exclusion of a repeating quartet). For example, one or more cysteine ​​residues may be incorporated into peptides and these residues may bind to each other by forming disulfide bonds.

Structures bonded in this way can have increased mechanical strength to structures made of comparable peptides that do not contain cysteine ​​residues and thus are unable to form disulfide bonds.

[0044] Amino acid residues in self-assembling peptides may be naturally occurring or non-naturally occurring amino acid residues. Naturally occurring amino acids may include amino acid residues coded with a standard genetic code, as well as non-standard amino acids (e.g., amino acids having a D configuration instead of an L configuration), as well as those amino acids that can be formed by modifications to standard amino acids (e.g., pyrolysine or selenocysteine). Naturally occurring amino acids are not found or not found in nature, but can be included in the peptide chain. Suitable non-naturally occurring amino acids include, but are not limited to, D-alloosoleucine, (2R, 3S) -2-amino-3-methylpentanoic acid, L-cyclopentylglycine, (S) -2-amino-2-cyclopentylacetic acid. Other examples of non-naturally occurring amino acids can be found in textbooks or on the web (e.g. a site run by the California Institute of Technology, which presents the structures of non-naturally occurring amino acids that have been successfully incorporated into functional proteins). Unnatural amino acid residues and amino acid derivatives are described in published US Patent Application No. 2004/0204561, Ellison.

[0045] Self-assembling peptides can be synthesized chemically or purified from natural or recombinantly produced sources by methods well known in the art. For example, peptides can be synthesized using standard chemical reactions using f-moc and purified using high pressure liquid chromatography (HPLC).

[0046] Self-assembling peptides such as EAKA16-I (SEQ ID No. 410), RADA16-I (SEQ ID No. 1), RAEA16-I (SEQ ID NO: 58) and KADA16-I (SEQ No. 59) is described in Zhang, S., et al. ((1999) Peptide self-assembly in functional polymer science and engineering. Reactive &amp; Functional Polymers, 41, 91-102). Self-assembling peptides comprise a sequence of amino acid residues corresponding to one or more of formulas I-IV:

<td></td><td>((Xaa^neu-Xaa⁺) X (Xaa^neu-Xaa^-) Y) n</td><td>(AND)</td>

<td>thirty</td><td>(Xaa^neu-Xaa^-) X (Xaa^neu-Xaa⁺) Y) n</td><td>(II)</td>

<td></td><td>((Xaa⁺-Xaa^neu) X (Xaa^--Xaa^neu) Y) n</td><td>(III)</td>

<td></td><td>((Xaa^--Xaa^neu) X (Xaa⁺-Xaa^neu) Y) n</td><td>(IV)</td>

Xaa^neu means an amino acid residue having an inert charge; Xaa⁺ is an amino acid residue having a positive charge; Xaa^- is an amino acid residue having a negative charge; x and y are integers having a value of 1, 2, 3 or 4 independently; and n is an integer of 1-5. The peptides of the I-module (i.e. peptides with alternately positive and negatively charged R groups on one side (e.g., polar β-sheet surface) are described by each of the formulas I-IV, in which x and y are 1. Peptides of the II module ( i.e. peptides having two residues carrying one type of charge (i.e., a positive charge) followed by two residues carrying a different type of charge (e.g., negative charge) are described using the same formulas in which both x and y are 2. Examples of module III peptides (i.e. peptides having three residues carrying one type of charge (e.g., a positive charge) followed by three residues carrying a different type of charge (e.g., negative charge)) include, but are not limited to,

[0047] Other hydrophilic moieties that form hydrogen bonds, including, but not limited to, asparagine and glutamine, can be incorporated into peptides. If the alanine residues in the peptides are changed to more hydrophobic residues, such as leucine, isoleucine, phenylalanine or tyrosine, the obtained peptides have a greater tendency to self-assemble and form peptide matrices with increased strength. Some peptides with similar amino acid sequences and lengths, such as the peptides described herein, form alpha-helixes and statistical-type structures, not beta-sheets, and do not form macroscopic structures. Thus, apart from self-complementarity, other factors may be important for the formation of macroscopic structures, such as the length of the peptide, the degree of intermolecular interaction and the ability to form alternate systems.

[0048] The peptide-based structures may be formed from heterogeneous mixtures of peptides (i.e. mixtures comprising more than one type of peptide corresponding to a given formula or two or more patterns). In some embodiments, each type of peptide in the mixture is capable of self-assembly. In other embodiments, one or more of each type of peptide would not self-assemble itself, but the combination of heterogeneous peptides may self-assemble (i.e., the peptides in the mixture are complementary and structurally compatible with each other). A homogeneous mixture of self-complementary and self-compatible peptides of the same sequence or containing the same recurring subunit, or a heterogeneous mixture of different peptides, can therefore be used,

[0049] In a preferred embodiment, a homogeneous or heterogeneous mixture of amino acid sequences that are not self-assembling can be added with one or more short amino acid sequences that aid self-assembly (referred to as assembly-assisting sequences). The assembly-assisting sequences may contain amino acids that are complementary to the amino acids in the sequences in the mixture. The assembly-assisting sequences may contain any number of amino acids. Preferably, the assembly-supporting sequences contain at least 4 amino acids. The assembly-assisting sequences may include a flexible link between amino acids that promotes self-assembly. For example, the assembly-assisting sequence may include a pair, triad or quartet of amino acids to assist assembly at the ends of the sequence, which are connected by a flexible connector. Suitable assembly-assisting sequences include, but are not limited to, COUNCIL (SEQ ID NO: 57) and EAKA (SEQ ID NO: 1111).

[0050] Suitable linkers include, but are not limited to, ether-based chains such as polyethylene glycol (PEG), N-succinimidyl 3- (2-pyridyldithio) propionate (SPDP, 3- and 7-atom insertion), long-chain SPDP ( 12-atom insert) (succinimidylcarbonyl-a-methyl-2- (2-pyridyl-dimethyl) toluene) (SMPT, 8-atom insert), succinimidyl 4- (N-maleimidomethyl) cyclohexane-1-carboxylate) (SMCC, insert 11- atomic) and sulfosuccinimidyl 4- (N-maleimidomethyl) cyclohexane-1-carboxylate (sulfo-SMCC, 11-atom insert), m-maleimidobenzoyl-N-hydroxysuccinimine ester (MBS, 9-atom insert)), N- ester ( Y-maleimidobutyryloxy) succinimide (GMBS, 8-atom insert), N- (Y-maleimidobutyryloxy) sulfosuccinimide ester (sulfo-GMBS, 8-atom insertion), succinimidyl 6 ((iodoacetyl) amino) hexanoate (SIAX, insert 9- atomic)Succinimidyl 6- (6 - (((4-iodoacetyl) amino) hexanoyl) amino) hexanoate (SIAXX, 16-atom insert) and p-nitrophenyl iodoacetate (NPIA, insertion

2-Nuclear). One of ordinary skill in the art should also note that many other linkers with different numbers of atoms can be used.

[0051] The compositions described herein independently of the exact form (e.g. whether in liquid or molded form) and independently of the general compositions (e.g., combined with another agent contained in the device or packaged in a kit) may contain a mixture of one or more the number of peptide chains.

[0052] Self-assembling structures that have different degrees of stiffness or elasticity may be formed. These structures usually have a small modulus of elasticity (e.g., a modulus in the range of about 0.01 to about 1000 kPa, preferably from about 1 to about 100 kPa, more preferably from about 1 to about 10 kPa measured by standard methods, such as using a standard rheometer cone-plate type). Low values ​​may be advantageous, because they allow deformation of the structure as a result of movement, in response to pressure, in the case of cell contractions. More specifically, the stiffness can be controlled in various ways, including by changing the length, sequence and / or concentration of precursor molecules (i.e., self-assembling peptides). Other methods of increasing stiffness may also be used. For example, biotin molecules or other molecules may be attached to the precursors, which may then be cross-linked or otherwise associated with each other. Molecules (e.g., biotin) may be included at the N- or C-terminus of the peptide or attached to one or more residues between the ends. Where biotin is used, cross-linking can be achieved by adding avidin again. Peptides containing biotin or peptides containing other molecules capable of crosslinking are included within the scope of the present invention. For example, amino acid residues with polymerizable groups, including, but not limited to, vinyl groups, can be introduced and cross-linked by irradiation with UV light. The degree of cross-linking can be precisely controlled by applying radiation for a predetermined time. The degree of crosslinking can be determined by light scattering, gel filtration or scanning electron microscopy using methods well known in the art. In addition, the cross-linking may be tested by HPLC or by mass spectrometry analysis of the protease digested mass structure such as matrix metalloproteases. Material strength can be determined before and after crosslinking. Regardless of whether crosslinking is achieved using a chemical agent or light energy, the molecules can be crosslinked during mold formation or when solutions containing the peptide are applied to the body. In addition, self-assembling peptide chains can be cross-linked to form a spider-like pattern to enhance the material in vivo. Transverse ties are used to strengthen the material, providing greater stiffness and durability. For example, self-assembling peptides can be applied to a wound, wherein the periphery of self-assembling peptides is functionalized by polymerizable groups. After cross-linking, the periphery of the self-assembling peptides becomes stiffer, anchoring the peptides at the wound site, while the inner part of the self-assembling peptides remains flexible to move with the movement of the organism.

[0053] The half-life (e.g., in vivo half-life) of the structures may also be modulated by including cleavage sites for proteases or peptidases into precursors that then form the given structure. Proteases or peptidases that occur naturally in vivo, or that are introduced (e.g., by a surgeon), can then promote degradation by cleaving their related substrates.

[0054] Combinations of any modifications described herein may be made. For example, self-assembling peptides may be used that contain a protease cleavage site and a cysteine ​​residue and / or a crosslinking agent, kits and devices containing them and methods of using them.

[0055] Peptide structures formed from any self-assembling peptides produced by any method can be characterized using various biophysical and optical techniques such as circular dichroism (CD), dynamic light scattering, Fourier transform infrared spectroscopy (FTIR), microscopy atomic forces (voltages) (ATM), scanning electron microscopy (SEM) and transmission electron microscopy (TEM). For example, biophysical methods can be used to determine the degree of secondary beta-sheet structure in the peptide structure. The size of fibrils and pores, fiber diameter, length, elasticity and fraction volume can be determined using quantitative analysis of the scanning image and / or transmission electron microscope. The structures can also be tested using several standard mechanical test techniques to measure the degree of swelling, the effect of pH and ion concentration on structure formation, the level of hydration under different conditions, tensile strength as well as the way in which different characteristics change over the time required to create and degrade structures. These methods allow one of ordinary skill in the art to determine which of the various alternatives and peptides described herein are the most suitable for use in various methods and to optimize various methods. how different characteristics change over the time required to create and degrade structures. These methods allow one of ordinary skill in the art to determine which of the various alternatives and peptides described herein are the most suitable for use in various methods and to optimize various methods. how different characteristics change over the time required to create and degrade structures. These methods allow one of ordinary skill in the art to determine which of the various alternatives and peptides described herein are the most suitable for use in various methods and to optimize various methods.

[0056] In another embodiment, the self-assembling peptides may anchor or interact with the structural extracellular matrix (ECM) at the borders of the blood vessels and / or tissues that are described herein. These self-assembling peptides typically have hydrophobic and / or hydrophilic sections that allow the material to react or interact with glycoproteins found in the ECM.

[0057] Preferably, self-assembling peptides do not cause secondary toxicity during disintegration. In addition, the decomposition product of self-assembling peptides is suitable for growth and repair of surrounding tissues.

1. Other self-accumulating materials [0058] Another embodiment provides self-assembling peptides having a segment of residues having a positive charge under physiological conditions connected to a segment of residues having a negative charge under physiological conditions. The segment of positively or negatively charged residues may comprise from about 2 to about 50 amino acid residues, typically from about 3 to about 30 residues, more usually from about 10 to about 20 amino acid residues. In another embodiment, about half the residues of the self-assembling peptide are positively charged and the other half of the self-assembling peptide have negatively charged amino acid residues. The combination of these peptides may self-assemble by overlapping the positive end of the first self-assembling peptide with the negative end of the second self-assembled peptide. The negative end of the first self-assembling peptide will overlap or align with the positive end of the second self-assembled peptide. Self-assembling peptides will aggregate or aggregate based on the opposite ends of self-assembling peptides that are attacked based on the charge in physiological compositions. One representative form provides a self-assembling peptide with the following RRRR sequence -DDDD (SEQ ID NO: 114) or GGGG-SSSS (SEQ ID NO: 115). [0059] In yet another embodiment, the self-assembling peptide has a first hydrophobic region operably linked to the first hydrophilic region. The first hydrophobic region can include a segment of amino acid residues that have hydrophobic side chains under physiological conditions. The first hydrophilic region can comprise a segment of amino acid residues that have hydrophilic side chains under physiological conditions. In this embodiment, the hydrophobic ends of the self-assembling peptides are folded together with other hydrophobic ends, and the hydrophilic ends are folded with other hydrophilic ends. Assembly can be controlled by changing the peptide environment. Such materials could be used to cover the interior of the light. The hydrophobic ends could possibly interact with the ECM of the light surface, sealing the surface while the hydrophilic ends extend towards the center of the light. Fluids could still flow through the light. As self-assembling peptides are degraded and / or removed from the surface of the light, the peptides will flow out of other areas and re-anchor in the surface of the light, and thus the composition acts as a reservoir supplying new peptides if necessary. Alternatively, additional self-assembling peptides may be administered to replace peptides that have been consumed or degraded. In another embodiment, self-assembling peptides may be used as dynamic slices, for example in the treatment of ulcers or for use in the intestines.

[0060] Another embodiment provides a self-assembling peptide that has a segment of residues that have a positive or negative charge under physiological conditions. Representative amino acid sequences of positively charged self-assembling peptides include, but are not limited to, KKKK (SEQ ID NO: 116), RRRR (SEQ ID NO: 117) or HHHH (SEQ ID

NO: 118). Representative amino acid sequences of negatively charged self-assembling peptides include, but are not limited to, DDDD (SEQ ID NO: 119) or EEEE (SEQ ID NO: 120). After combining, a series of positively charged amino acid residues will be aligned in parallel and opposite to a series of negatively charged amino acid residues. In certain embodiments, the series of positively charged amino acids will alternate with the series of negatively charged amino acids in a multilamellar structure.

[0061] Yet another embodiment provides self-assembling peptides that have a combination of hydrophilic polar amino acid residues and hydrophobic non-polar amino acid residues under physiological conditions. One or more hydrophilic residues may alternate with one or more hydrophobic residues. For example, the amino acid sequence of a representative self-assembling peptide may be GQGQ (SEQ ID NO: 121), GGQQGG (SEQ ID NO: 122), GQQGQQG (SEQ ID NO: 123), GGQGGQGG (SEQ ID NO: 124), etc. It should be noted that the division of the self-assembling peptide into a polar or non-polar environment can be controlled by changing the ratio of the hydrophobic amino acid residues to the hydrophilic amino acid residues, wherein a ratio greater than 1: 1 indicates that that the partition of peptide fragments takes place to a greater extent under hydrophobic conditions compared to hydrophilic conditions. A ratio of less than 1: 1 indicates that the partition of the peptide fragments takes place to a greater extent under hydrophilic conditions compared to hydro-phobic conditions.

[0062] Any of the modifications described herein can be made. For example, self-assembling peptides may be used that contain a protease cleavage site and a cysteine ​​residue and / or a crosslinking agent, kits and tools containing them, and methods of using them. The compositions may be used to prevent or limit the flow of body fluid, stabilize tissue or cells, or prevent contamination upon administration to a demanding site. The compositions may be in the form of a dry powder, wafer, disk, tablets, capsules, liquid, gel, cream, foam, ointment, emulsion, stent coating, catheter or other medical implant, peptides inserted into the microparticle, polymer matrix, hydrogel, fabric, bandage , seam or sponge.

B. Formation of self-assembling peptides Prior to self-assembly, self-assembling peptides may be (e.g., dissolved) in a solution that is essentially free of ion (e.g., monovalent ion) or that contains a sufficiently low ion concentration to prevent significant self-depositing (e.g., ion concentrations below 10, 5, 1 or 0.1 mM). Self-assembly can be initiated or enhanced at any later time by adding an ionic solute or diluent to the material solution or by changing the pH. For example, NaCl in a concentration between about 5 mM and 5 M may induce the assembly of macroscopic structures in a short period of time (e.g., within a few minutes). Lower NaCl concentrations can also induce assembly, although at lower rates. Possibly, self-assembly may be initiated or enhanced by the introduction of peptides (in a dry form, in a semi-solid gel or dissolved in a liquid solution that is essentially ion-free) to a fluid (e.g., physiological liquid such as blood or gastric juice) or area (e.g. a body cavity such as a nose or mouth, or a cavity exposed in a surgical procedure) containing such ions. The gel does not have to be pre-formed before applying to the desired place. In general, self-assembly should occur in any way after the peptides are contacted with such a solution. or a cavity exposed in a surgical procedure) containing such ions. The gel does not have to be pre-formed before applying to the desired place. In general, self-assembly should occur in any way after the peptides are contacted with such a solution. or a cavity exposed in a surgical procedure) containing such ions. The gel does not have to be pre-formed before applying to the desired place. In general, self-assembly should occur in any way after the peptides are contacted with such a solution.

[0064] A variety of ions may be used, including anions and cations (both divalent, monovalent and trivalent). For example, a phase transition can be achieved by exposure to monovalent cations such as Li⁺, Na⁺.

K⁺ and Cs⁺. The concentration of such ions needed to induce or enhance self-assembly is usually at least 5 mM (e.g., at least 10, 20 or 50 mM). Lower concentrations also facilitate folding, although at lower speeds. If desired, self-assembling peptides can be provided with a hydrophobic material (e.g., a pharmaceutically acceptable oil) at a concentration that allows self-assembly, albeit at a lower rate. When the self-assembling peptides are mixed with a hydrophobic agent, such as an oil or lipid, the assembly of the peptides leads to the formation of various structures. The structures will look like ice on the oil layer. In some cases, when another material is added, the material will be folded to form various other three-dimensional structures that may be suitable for loading with a therapeutic agent.riera between two environments. Several experiments have shown that self-assembling peptides will align on the surface of the oil, like ice on water, with the hydrophobic part of the molecule towards the surface and the hydrophilic part of the molecule in the opposite direction to the oil or form toroidal structures with the hydrophobic material contained within. This type of behavior will allow the encapsulation of therapeutic agents or other molecules of interest to be delivered to the body.

[0065] In another embodiment, the composition may include a salt scavenger to direct assembly into a preferred configuration. For example, circular dichroism experiments ("CDs") indicate that the folding dynamics can be controlled by scavengers in the form of salts or enhancers in the form of salts to increase the formation of β-sheets, α-helices or more random configurations. The compositions may optionally include an indicator indicative of the assembly configuration (e.g., α-helix, β-sheet, meshes, etc.).

[0066] Optionally, some of the self-assembling peptides described herein do not require self-assembly ions, but may self-assemble due to interaction with the solvent, hydrophobic interactions, side-chain interactions, hydrogen bonding and the like.

[0067] Depending on the composition and desired properties of the macroscopic structure (e.g.

stiffness or rate of formation), the concentration of precursors (i.e., self-assembling peptides) may vary from about 0.01% w / v. (0.1 mg / ml) to about 99.99% w / v. (999.9 mg / ml), inclusive. For example, the concentration prior to forming the scaffold may be between about 0.1% (1 mg / mL) and 10% (100 mg / mL), inclusive (e.g., about 0.1% -5%, 0.5% -5%; 1.0%, 1.5%, 2.0%, 2.5%, 3.0% or 4.0%, or more). Precursors (i.e., self-assembling peptides) may be formulated as powders and administered as a powder or resuspended. If they are in a dry form, the peptides may then self-assemble upon contact with body fluids (e.g., in the wound site).

[0068] Self-assembling peptides may be formed in regular or irregular forms, which may include the body cavity or body part (e.g., the lumen of a blood vessel), or which may be an inert material such as plastic or glass. The structures or scaffolds can be made to correspond to a predetermined shape or have a predetermined volume. In order to form a structure with a predetermined shape or volume (e.g., with a desired geometry or dimension, including thin sheets or film), an aqueous solution of self-assembling peptides is placed in a preformed mold for casting and peptide self-assembly is triggered by the addition of multiple ions. . Alternatively, the ions may be added to the solution shortly before the solution is placed in the mold, provided that it is taken into account to put the solution in the mold before the main folding event. If the form is tissue (e.g., blood vessel light or other compartment, in situ or not), the addition of the ionic solution may not be necessary. The resulting characteristics of self-assembling peptides, the time needed for assembly and the dimensions of the macroscopic structure that is formed are controlled by the concentration and amount of solution used, the concentration of ions used to induce assembly of the structure, and the dimensions of the casting device. The scaffold may reach a gel or substantially solid form at room temperature and heat may be provided to facilitate molding (e.g., the solution used in the molding process (e.g., precursor-containing solution) may be heated to a temperature approaching approximately body temperature (about 37 ° C)) . If the form is tissue (e.g., blood vessel light or other compartment, in situ or not), the addition of the ionic solution may not be necessary. The resulting characteristics of self-assembling peptides, the time needed for assembly and the dimensions of the macroscopic structure that is formed are controlled by the concentration and amount of solution used, the concentration of ions used to induce assembly of the structure, and the dimensions of the casting device. The scaffold may reach a gel or substantially solid form at room temperature and heat may be provided to facilitate molding (e.g., the solution used in the molding process (e.g., precursor-containing solution) may be heated to a temperature approaching approximately body temperature (about 37 ° C)) . If the form is tissue (e.g., blood vessel light or other compartment, in situ or not), the addition of the ionic solution may not be necessary. The resulting characteristics of self-assembling peptides, the time needed for assembly and the dimensions of the macroscopic structure that is formed are controlled by the concentration and amount of solution used, the concentration of ions used to induce assembly of the structure, and the dimensions of the casting device. The scaffold may reach a gel or substantially solid form at room temperature and heat may be provided to facilitate molding (e.g., the solution used in the molding process (e.g., precursor-containing solution) may be heated to a temperature approaching approximately body temperature (about 37 ° C)) . adding an ionic solution may not be necessary. The resulting characteristics of self-assembling peptides, the time needed for assembly and the dimensions of the macroscopic structure that is formed are controlled by the concentration and amount of solution used, the concentration of ions used to induce assembly of the structure, and the dimensions of the casting device. The scaffold may reach a gel or substantially solid form at room temperature and heat may be provided to facilitate molding (e.g., the solution used in the molding process (e.g., precursor-containing solution) may be heated to a temperature approaching approximately body temperature (about 37 ° C)) . adding an ionic solution may not be necessary. The resulting characteristics of self-assembling peptides, the time needed for assembly and the dimensions of the macroscopic structure that is formed are controlled by the concentration and amount of solution used, the concentration of ions used to induce assembly of the structure, and the dimensions of the casting device. The scaffold may reach a gel or substantially solid form at room temperature and heat may be provided to facilitate molding (e.g., the solution used in the molding process (e.g., precursor-containing solution) may be heated to a temperature approaching approximately body temperature (about 37 ° C)) . are controlled by the concentration and amount of solution used, the concentration of ions used to induce assembly of the structure, as well as the dimensions of the casting device. The scaffold may reach a gel or substantially solid form at room temperature and heat may be provided to facilitate molding (e.g., the solution used in the molding process (e.g., precursor-containing solution) may be heated to a temperature approaching approximately body temperature (about 37 ° C)) . are controlled by the concentration and amount of solution used, the concentration of ions used to induce assembly of the structure, as well as the dimensions of the casting device. The scaffold may reach a gel or substantially solid form at room temperature and heat may be provided to facilitate molding (e.g., the solution used in the molding process (e.g., precursor-containing solution) may be heated to a temperature approaching approximately body temperature (about 37 ° C)) .

When the scaffolding reaches the desired degree of hardness, it can be removed from the mold and used for the purpose described here. Alternatively, the self-assembling peptides described herein can be used to anchor host tissue in a matrix or tissue scaffold. For example, the self-assembling peptides described herein can be used as an adhesive for anchoring a host tissue to be regenerated in a matrix or tissue scaffold to ensure that the matrix or scaffolding remains immobile in the local environment into which they are injected or implanted. Dental ties and scaffolds are well known in the art and can be made from synthetic, semi-synthetic and / or natural materials.

[0069] Peptides that undergo folding and / or phase transition (e.g., transition from liquid to semi-solid state, gel, etc.) when in contact with the body or ionic solution are useful in preventing the movement of body substances. Self-assembly or phase transition is triggered by components occurring in the body of the subject (e.g., ions) or by physiological pH and is aided by physiological temperatures. Self-assembly or phase transition can begin when the compositions are exposed to or coming into contact with the subject's body and can be facilitated by local application of heat to the area in which the composition has been (or will be) deposited. Based on previous studies, self-assembly occurs quickly after contact with internal body tissues without the use of additional heat. The time required for effective folding and / or phase transition may occur within 60 seconds or less after contact with the internal tissues of the subject or in conditions similar to those found in the body (e.g., within 50, 40, 30, 20 or 10 seconds) or less). In some circumstances, for example where the concentration of self-assembling peptides in the composition is low, or when the body's movement is significant, self-assembly or phase transition may take longer to achieve the desired effect, for example, up to a minute, 5 minutes , 10 minutes, 30 minutes, one hour or more. For example, a solution containing a self-assembled peptide applied at sites of blood vessel incisions in the brain, liver or muscles provided complete haemostasis within only 10 seconds after application.

[0070] The compositions may form structures that are substantially rigid (e.g., solid or nearly constant) or that take a specific shape and volume (e.g., structures that correspond to the shape and volume of the site to which the liquid composition is administered, whether in vivo or ex vivo). The solidified material may be somewhat deformable or compressible after folding or phase transition, but it will not essentially flow from one region to another, such as compositions at another point on the continuum of the liquid to the solid, which may be dependent, at least in part, on their phase transition capability. As a result, the compositions can be used to prevent the movement of the body substance in a subject in need thereof. Self-assembly can be achieved in vivo or ex vivo by exposure to conditions within a range of physiological values ​​(e.g., conditions suitable for cell or tissue culture) or non-physiological conditions. "Non-physiological conditions" refer to conditions in the body or in a particular place that deviate from the normal physiological conditions in this place. Such conditions may result from injury, surgery, wounding, infection or disease, disorder or condition. For example, a stab wound in the stomach usually causes a drop in pH because gastric acid flows to the wound site. The peptides described herein should undergo self-assembly under such conditions. While liquid formulations are readily dosed, the administered compositions may also be in the form of a gel that may become stiff after contact with the subject's body. conditions suitable for cell or tissue culture) or non-physiological conditions. "Non-physiological conditions" refer to conditions in the body or in a particular place that deviate from the normal physiological conditions in this place. Such conditions may result from injury, surgery, wounding, infection or disease, disorder or condition. For example, a stab wound in the stomach usually causes a drop in pH because gastric acid flows to the wound site. The peptides described herein should undergo self-assembly under such conditions. While liquid formulations are readily dosed, the administered compositions may also be in the form of a gel that may become stiff after contact with the subject's body. conditions suitable for cell or tissue culture) or non-physiological conditions. "Non-physiological conditions" refer to conditions in the body or in a particular place that deviate from the normal physiological conditions in this place. Such conditions may result from injury, surgery, wounding, infection or disease, disorder or condition. For example, a stab wound in the stomach usually causes a drop in pH because gastric acid flows to the wound site. The peptides described herein should undergo self-assembly under such conditions. While liquid formulations are readily dosed, the administered compositions may also be in the form of a gel that may become stiff after contact with the subject's body. "Non-physiological conditions" refer to conditions in the body or in a particular place that deviate from the normal physiological conditions in this place. Such conditions may result from injury, surgery, wounding, infection or disease, disorder or condition. For example, a stab wound in the stomach usually causes a drop in pH because gastric acid flows to the wound site. The peptides described herein should undergo self-assembly under such conditions. While liquid formulations are readily dosed, the administered compositions may also be in the form of a gel that may become stiff after contact with the subject's body. "Non-physiological conditions" refer to conditions in the body or in a particular place that deviate from the normal physiological conditions in this place. Such conditions may result from injury, surgery, wounding, infection or disease, disorder or condition. For example, a stab wound in the stomach usually causes a drop in pH because gastric acid flows to the wound site. The peptides described herein should undergo self-assembly under such conditions. While liquid formulations are readily dosed, the administered compositions may also be in the form of a gel that may become stiff after contact with the subject's body. the stab wound in the stomach usually causes a drop in pH, because gastric acid flows to the site of the wound. The peptides described herein should undergo self-assembly under such conditions. While liquid formulations are readily dosed, the administered compositions may also be in the form of a gel that may become stiff after contact with the subject's body. the stab wound in the stomach usually causes a drop in pH, because gastric acid flows to the site of the wound. The peptides described herein should undergo self-assembly under such conditions. While liquid formulations are readily dosed, the administered compositions may also be in the form of a gel that may become stiff after contact with the subject's body.

[0071] The concentration of self-assembling peptides in any given formulation may vary and may range from about 0.1% (1 mg / mL) and 10% (100 mg / mL), inclusive. For example, the concentration of self-assembling peptides (e.g., in a liquid formulation) may be about 0.1-3.0% (1-30 mg / mL) (e.g., 0.1-1.0%, 1.0-2, 0%, 2.0-3.0% or 1.0-3.0%). The concentration of self-assembling peptides may be higher in the main solutions and in solid preparations (e.g. powdered). In solid preparations, the concentration of self-assembling peptides may approach 100% (e.g., the concentration of self-assembling peptides may be 95, 96, 97, 98, 99% or more (e.g. 99.99%) of the composition). Regardless of whether they are in a liquid or solid form, the self-assembling peptides can be brought to the desired concentration before use by adding a diluent (e.g.

Regardless of the exact nature of the self-assembling peptides, upon exposure to conditions such as those described herein, the peptides can form membrane bi- or three-dimensional structures, including a stable macroscopic porous matrix having ordered or disordered interlaced nanofibres (e.g., fibers with a diameter). about 10-20 nm, with a pore size of about 50-100 nm in the linear dimension). The three-dimensional macroscopic matrices may have dimensions large enough to be visible at low magnification (e.g., about 10-fold or less), and the membrane structures may be visible to the naked eye, even if they are transparent. Although they are three-dimensional, structures can be very thin, including a limited number of molecular layers (e.g., 2, 3 or more layers of molecules). Typically, each dimension of a given structure will have a size of at least 10 μm (e.g., two dimensions of at least 100-1000 μm (e.g., 1-10 mm, 10-100 mm or more)). Suitable dimensions can be expressed as length, width, depth, span, height, radius, diameter or circumference in the case of structures that have a substantially regular shape (e.g., when the structure is a ball, cylinder, cube or the like) or approximation of any of the above when the structures do not have a regular shape.

[0073] Self-assembling peptides can form a hydrated material upon contact with water under conditions such as those described herein (e.g., in the presence of a sufficient concentration (e.g., physiological concentrations) of ions (e.g., monovalent cations)). Self-assembling peptides can have a high water content (e.g., about 95% or more (e.g., about 97%, 98%, 99% or more)) and the compositions can be hydrated, but are generally non-self-depositing. The given value may be "approximate" taking into account the fact that the measurements may vary depending, for example, on the circumstances in which they are made and the skills of the person performing the measurement. In general, the first value is approximately equal to the second when the former is within 10% of the second (whether greater or smaller than), unless it is clear from the context that the value is not approximate or when, for example,

[0074] The properties and mechanical strength of the structures or scaffolds can be controlled as required by manipulating their components. For example, the stiffness of a submissive gel assembly can be increased by increasing the concentration of self-assembling peptides. Alternatively, it may be desirable for various portions of self-assembling peptides to have different mechanical properties. For example, it may be advantageous to reduce the stability of all or part of self-assembling peptides by manipulating the amino acid sequence. This may be desirable when the self-assembling peptides are used to fill the void so that the peptide edges self-assemble to attach to the tissue site while the remaining peptides flow into the void space. sequences

[0075] The compositions may be formulated in concentrated stock solutions or in a dry form and may be diluted or dissolved to produce biocompatible compositions that are substantially non-toxic to biological cells in vitro or in vivo. For example, the compositions may contain materials in amounts that do not cause significant deleterious effects to the recipient (e.g., a disproportionately severe immune or inflammatory reaction, or unacceptable scar tissue formation).

[0076] When a solution containing non-foldable peptides is found on the biological tissue, the peptides being sufficiently close to the tissue fold, resulting in gelation of the solution. Any part of the solution remaining far from the tissue remains fluid, because the self-assembling peptides have not yet been exposed to the conditions favoring their assembly. Because self-assembling peptides are disrupted (e.g., by a surgical procedure), it appears that the liquid material gels when it comes into contact with the body sufficiently. Occasionally, the compositions may take on characteristics ranging from liquid to solid, looking similar to a gel or ointment, or as a suspension.

C. Modification of self-assembled materials for specific target tissues [0077] Self-assembling peptides further comprise a tissue-specific component. The tissue-specific component may be peptides that are specific to the cells of the eye, brain or skin. For example, cell surface carbohydrates are the major components of the outer surface of mammalian cells and are very often characteristic of cell types. It is assumed that carbohydrates specific for the cell type are involved in the cell-cell interaction. The tissue-specific component may therefore target these cell-specific surface carbohydrates.

[0078] In addition, hydrophobic tails may be added to self-assembling peptides. Tails can interact with cell membranes, which causes anchoring of self-assembling peptides on the cell surface. Table 3 lists the peptides with hydrophobic tails. The hydrophilic tails can also be added to the peptides, in addition to the hydrophobic tails, to facilitate interaction with the ECM of various vessels or tissues, such as the bladder.

Table 3. Hydrophobic tails

<td>1</td><td>G</td><td>G</td><td>G</td><td>G</td><td>G</td><td>D</td><td>G</td><td>D</td><td>G</td><td>D</td><td>G</td><td>D</td><td>G</td><td>D</td><td>G</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>126)</td>

<td>2</td><td>G</td><td>G</td><td>G</td><td>G</td><td>G</td><td>E</td><td>G</td><td>E</td><td>G</td><td>E</td><td>G</td><td>E</td><td>G</td><td>E</td><td>G</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>127)</td>

<td>3</td><td>G</td><td>G</td><td>G</td><td>G</td><td>G</td><td>K</td><td>G</td><td>K</td><td>G</td><td>K</td><td>G</td><td>K</td><td>G</td><td>K</td><td>G</td><td>K</td><td>(SEQ.</td><td>ID NO.</td><td>128)</td>

<td>4</td><td>G</td><td>G</td><td>G</td><td>G</td><td>G</td><td>R</td><td>G</td><td>R</td><td>G</td><td>R</td><td>G</td><td>R</td><td>G</td><td>R</td><td>G</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>129)</td>

<td>5</td><td>G</td><td>G</td><td>G</td><td>G</td><td>G</td><td>H</td><td>G</td><td>H</td><td>G</td><td>H</td><td>G</td><td>H</td><td>G</td><td>H</td><td>G</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>130)</td>

<td>6</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>D</td><td>AND</td><td>D</td><td>AND</td><td>D</td><td>AND</td><td>D</td><td>AND</td><td>D</td><td>AND</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>131)</td>

<td>7</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>E</td><td>AND</td><td>E</td><td>AND</td><td>E</td><td>AND</td><td>E</td><td>AND</td><td>E</td><td>AND</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>132)</td>

<td>8</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>K</td><td>AND</td><td>K</td><td>AND</td><td>K</td><td>AND</td><td>K</td><td>AND</td><td>K</td><td>AND</td><td>K</td><td>(SEQ.</td><td>ID NO.</td><td>133)</td>

<td>9</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>134)</td>

<td>10</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>H</td><td>AND</td><td>H</td><td>AND</td><td>H</td><td>AND</td><td>H</td><td>AND</td><td>H</td><td>AND</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>135)</td>

<td>11</td><td>V</td><td>V</td><td>V</td><td>V</td><td>V</td><td>D</td><td>V</td><td>D</td><td>V</td><td>D</td><td>V</td><td>D</td><td>V</td><td>D</td><td>V</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>136)</td>

<td>12</td><td>V</td><td>V</td><td>V</td><td>V</td><td>V</td><td>E</td><td>V</td><td>E</td><td>V</td><td>E</td><td>V</td><td>E</td><td>V</td><td>E</td><td>V</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>137)</td>

<td>13</td><td>V</td><td>V</td><td>V</td><td>V</td><td>V</td><td>K</td><td>V</td><td>K</td><td>V</td><td>K</td><td>V</td><td>K</td><td>V</td><td>K</td><td>V</td><td>K</td><td>(SEQ.</td><td>ID NO.</td><td>138)</td>

<td>14</td><td>V</td><td>V</td><td>V</td><td>V</td><td>V</td><td>R</td><td>V</td><td>R</td><td>V</td><td>R</td><td>V</td><td>R</td><td>V</td><td>R</td><td>V</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>139)</td>

<td>15</td><td>V</td><td>V</td><td>V</td><td>V</td><td>V</td><td>H</td><td>V</td><td>H</td><td>V</td><td>H</td><td>V</td><td>H</td><td>V</td><td>H</td><td>V</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>140)</td>

<td>16</td><td>L</td><td>L</td><td>L</td><td>L</td><td>L</td><td>D</td><td>L</td><td>D</td><td>L</td><td>D</td><td>L</td><td>D</td><td>L</td><td>D</td><td>L</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>141)</td>

<td>17</td><td>L</td><td>L</td><td>L</td><td>L</td><td>L</td><td>E</td><td>L</td><td>E</td><td>L</td><td>E</td><td>L</td><td>E</td><td>L</td><td>E</td><td>L</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>142)</td>

<td>18</td><td>L</td><td>L</td><td>L</td><td>L</td><td>L</td><td>K</td><td>L</td><td>K</td><td>L</td><td>K</td><td>L</td><td>K</td><td>L</td><td>K</td><td>L</td><td>K</td><td>(SEQ.</td><td>ID NO.</td><td>143)</td>

<td>19</td><td>L</td><td>L</td><td>L</td><td>L</td><td>L</td><td>R</td><td>L</td><td>R</td><td>L</td><td>R</td><td>L</td><td>R</td><td>L</td><td>R</td><td>L</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>144)</td>

<td>20</td><td>L</td><td>L</td><td>L</td><td>L</td><td>L</td><td>H</td><td>L</td><td>H</td><td>L</td><td>H</td><td>L</td><td>H</td><td>L</td><td>H</td><td>L</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>145)</td>

<td>21</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>D</td><td>AND</td><td>D</td><td>AND</td><td>D</td><td>AND</td><td>D</td><td>AND</td><td>D</td><td>AND</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>146)</td>

<td>22</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>E</td><td>AND</td><td>E</td><td>AND</td><td>E</td><td>AND</td><td>E</td><td>AND</td><td>E</td><td>AND</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>147)</td>

<td>23</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>K</td><td>AND</td><td>K</td><td>AND</td><td>K</td><td>AND</td><td>K</td><td>AND</td><td>K</td><td>AND</td><td>K</td><td>(SEQ.</td><td>ID NO.</td><td>148)</td>

<td>24</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>149)</td>

<td>25</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>H</td><td>AND</td><td>H</td><td>AND</td><td>H</td><td>AND</td><td>H</td><td>AND</td><td>H</td><td>AND</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>150)</td>

<td>26</td><td>M</td><td>M</td><td>M</td><td>M</td><td>M</td><td>D</td><td>M</td><td>D</td><td>M</td><td>D</td><td>M</td><td>D</td><td>M</td><td>D</td><td>M</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>151)</td>

<td>27</td><td>M</td><td>M</td><td>M</td><td>M</td><td>M</td><td>E</td><td>M</td><td>E</td><td>M</td><td>E</td><td>M</td><td>E</td><td>M</td><td>E</td><td>M</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>152)</td>

<td>28</td><td>M</td><td>M</td><td>M</td><td>M</td><td>M</td><td>K</td><td>M</td><td>K</td><td>M</td><td>K</td><td>M</td><td>K</td><td>M</td><td>K</td><td>M</td><td>K</td><td>(SEQ.</td><td>ID NO.</td><td>153)</td>

<td>29</td><td>M</td><td>M</td><td>M</td><td>M</td><td>M</td><td>R</td><td>M</td><td>R</td><td>M</td><td>R</td><td>M</td><td>R</td><td>M</td><td>R</td><td>M</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>154)</td>

<td>thirty</td><td>M</td><td>M</td><td>M</td><td>M</td><td>M</td><td>H</td><td>M</td><td>H</td><td>M</td><td>H</td><td>M</td><td>H</td><td>M</td><td>H</td><td>M</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>155)</td>

<td>31</td><td>F</td><td>F</td><td>F</td><td>F</td><td>F</td><td>D</td><td>F</td><td>D</td><td>F</td><td>D</td><td>F</td><td>D</td><td>F</td><td>D</td><td>F</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>156)</td>

<td>32</td><td>F</td><td>F</td><td>F</td><td>F</td><td>F</td><td>E</td><td>F</td><td>E</td><td>F</td><td>E</td><td>F</td><td>E</td><td>F</td><td>E</td><td>F</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>157)</td>

<td>33</td><td>F</td><td>F</td><td>F</td><td>F</td><td>F</td><td>K</td><td>F</td><td>K</td><td>F</td><td>K</td><td>F</td><td>K</td><td>F</td><td>K</td><td>F</td><td>K</td><td>(SEQ.</td><td>ID NO.</td><td>158)</td>

<td>34</td><td>F</td><td>F</td><td>F</td><td>F</td><td>F</td><td>R</td><td>F</td><td>R</td><td>F</td><td>R</td><td>F</td><td>R</td><td>F</td><td>R</td><td>F</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>159)</td>

<td>35</td><td>F</td><td>F</td><td>F</td><td>F</td><td>F</td><td>H</td><td>F</td><td>H</td><td>F</td><td>H</td><td>F</td><td>H</td><td>F</td><td>H</td><td>F</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>160)</td>

<td>36</td><td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>D</td><td>IN</td><td>D</td><td>IN</td><td>D</td><td>IN</td><td>D</td><td>IN</td><td>D</td><td>IN</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>161)</td>

<td>37</td><td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>E</td><td>IN</td><td>E</td><td>IN</td><td>E</td><td>IN</td><td>E</td><td>IN</td><td>E</td><td>IN</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>162)</td>

<td>38</td><td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>K</td><td>IN</td><td>K</td><td>IN</td><td>K</td><td>IN</td><td>K</td><td>IN</td><td>K</td><td>IN</td><td>K</td><td>(SEQ.</td><td>ID NO.</td><td>163)</td>

<td>39</td><td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>R</td><td>IN</td><td>R</td><td>IN</td><td>R</td><td>IN</td><td>R</td><td>IN</td><td>R</td><td>IN</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>164)</td>

<td>40</td><td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>H</td><td>IN</td><td>H</td><td>IN</td><td>H</td><td>IN</td><td>H</td><td>IN</td><td>H</td><td>IN</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>165)</td>

<td>41</td><td>P</td><td>P</td><td>P</td><td>P</td><td>P</td><td>D</td><td>P</td><td>D</td><td>P</td><td>D</td><td>P</td><td>D</td><td>P</td><td>D</td><td>P</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>166)</td>

<td>42</td><td>P</td><td>P</td><td>P</td><td>P</td><td>P</td><td>E</td><td>P</td><td>E</td><td>P</td><td>E</td><td>P</td><td>E</td><td>P</td><td>E</td><td>P</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>167)</td>

<td>43</td><td>P</td><td>P</td><td>P</td><td>P</td><td>P</td><td>K</td><td>P</td><td>K</td><td>P</td><td>K</td><td>P</td><td>K</td><td>P</td><td>K</td><td>P</td><td>K</td><td>(SEQ.</td><td>ID NO.</td><td>168)</td>

<td>44</td><td>P</td><td>P</td><td>P</td><td>P</td><td>P</td><td>R</td><td>P</td><td>R</td><td>P</td><td>R</td><td>P</td><td>R</td><td>P</td><td>R</td><td>P</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>169)</td>

<td>45</td><td>P</td><td>P</td><td>P</td><td>P</td><td>P</td><td>H</td><td>P</td><td>H</td><td>P</td><td>H</td><td>P</td><td>H</td><td>P</td><td>H</td><td>P</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>170)</td>

<td>46</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>R</td><td>AND</td><td>D</td><td>AND</td><td>R</td><td>AND</td><td>D</td><td>AND</td><td>R</td><td>AND</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>171)</td>

<td>47</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>D</td><td>AND</td><td>D</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>172)</td>

<td>48</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>E</td><td>AND</td><td>K</td><td>AND</td><td>E</td><td>AND</td><td>K</td><td>AND</td><td>E</td><td>AND</td><td>K</td><td>(SEQ.</td><td>ID NO.</td><td>173)</td>

<td>49</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>E</td><td>AND</td><td>E</td><td>AND</td><td>K</td><td>AND</td><td>K</td><td>AND</td><td>E</td><td>AND</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>174)</td>

<td>50</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>R</td><td>AND</td><td>E</td><td>AND</td><td>R</td><td>AND</td><td>E</td><td>AND</td><td>R</td><td>AND</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>175)</td>

<td>51</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>E</td><td>AND</td><td>E</td><td>AND</td><td>R</td><td>AND</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>176)</td>

<td>52</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>K</td><td>AND</td><td>D</td><td>AND</td><td>K</td><td>AND</td><td>D</td><td>AND</td><td>K</td><td>AND</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>177)</td>

<td>53</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>E</td><td>AND</td><td>H</td><td>AND</td><td>E</td><td>AND</td><td>H</td><td>AND</td><td>E</td><td>AND</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>178)</td>

<td>54</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>E</td><td>AND</td><td>E</td><td>AND</td><td>H</td><td>AND</td><td>H</td><td>AND</td><td>E</td><td>AND</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>179)</td>

<td>55</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>180)</td>

<td>56</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>D</td><td>AND</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>181)</td>

<td>57</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>D</td><td>AND</td><td>D</td><td>AND</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>182)</td>

<td>58</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>H</td><td>AND</td><td>D</td><td>AND</td><td>H</td><td>AND</td><td>D</td><td>AND</td><td>H</td><td>AND</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>183)</td>

<td>59</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>H</td><td>AND</td><td>H</td><td>AND</td><td>H</td><td>AND</td><td>H</td><td>AND</td><td>H</td><td>AND</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>184)</td>

<td>60</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>H</td><td>AND</td><td>D</td><td>AND</td><td>D</td><td>AND</td><td>H</td><td>AND</td><td>D</td><td>AND</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>185)</td>

<td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>H</td><td>AND</td><td>E</td><td>AND</td><td>E</td><td>AND</td><td>H</td><td>AND</td><td>E</td><td>AND</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>186)</td>

<td>G</td><td>G</td><td>G</td><td>G</td><td>G</td><td>R</td><td>G</td><td>D</td><td>G</td><td>R</td><td>G</td><td>D</td><td>G</td><td>R</td><td>G</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>187)</td>

<td>G</td><td>G</td><td>G</td><td>G</td><td>G</td><td>R</td><td>G</td><td>R</td><td>G</td><td>D</td><td>G</td><td>D</td><td>G</td><td>R</td><td>G</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>188)</td>

<td>G</td><td>G</td><td>G</td><td>G</td><td>G</td><td>E</td><td>G</td><td>K</td><td>G</td><td>E</td><td>G</td><td>K</td><td>G</td><td>E</td><td>G</td><td>K</td><td>(SEQ.</td><td>ID NO.</td><td>189)</td>

<td>G</td><td>G</td><td>G</td><td>G</td><td>G</td><td>E</td><td>G</td><td>E</td><td>G</td><td>K</td><td>G</td><td>K</td><td>G</td><td>E</td><td>G</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>190)</td>

<td>G</td><td>G</td><td>G</td><td>G</td><td>G</td><td>R</td><td>G</td><td>E</td><td>G</td><td>R</td><td>G</td><td>E</td><td>G</td><td>R</td><td>G</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>191)</td>

<td>G</td><td>G</td><td>G</td><td>G</td><td>G</td><td>R</td><td>G</td><td>R</td><td>G</td><td>E</td><td>G</td><td>E</td><td>G</td><td>R</td><td>G</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>192)</td>

<td>G</td><td>G</td><td>G</td><td>G</td><td>G</td><td>K</td><td>G</td><td>D</td><td>G</td><td>K</td><td>G</td><td>D</td><td>G</td><td>K</td><td>G</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>193)</td>

<td>G</td><td>G</td><td>G</td><td>G</td><td>G</td><td>E</td><td>G</td><td>H</td><td>G</td><td>E</td><td>G</td><td>H</td><td>G</td><td>E</td><td>G</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>194)</td>

<td>G</td><td>G</td><td>G</td><td>G</td><td>G</td><td>E</td><td>G</td><td>E</td><td>G</td><td>H</td><td>G</td><td>H</td><td>G</td><td>E</td><td>G</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>195)</td>

<td>G</td><td>G</td><td>G</td><td>G</td><td>G</td><td>R</td><td>G</td><td>R</td><td>G</td><td>R</td><td>G</td><td>R</td><td>G</td><td>R</td><td>G</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>196)</td>

<td>G</td><td>G</td><td>G</td><td>G</td><td>G</td><td>R</td><td>G</td><td>R</td><td>G</td><td>R</td><td>G</td><td>R</td><td>G</td><td>D</td><td>G</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>197)</td>

<td>G</td><td>G</td><td>G</td><td>G</td><td>G</td><td>R</td><td>G</td><td>R</td><td>G</td><td>R</td><td>G</td><td>D</td><td>G</td><td>D</td><td>G</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>198)</td>

<td>G</td><td>G</td><td>G</td><td>G</td><td>G</td><td>H</td><td>G</td><td>D</td><td>G</td><td>H</td><td>G</td><td>D</td><td>G</td><td>H</td><td>G</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>199)</td>

<td>G</td><td>G</td><td>G</td><td>G</td><td>G</td><td>H</td><td>G</td><td>H</td><td>G</td><td>H</td><td>G</td><td>H</td><td>G</td><td>H</td><td>G</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>200)</td>

<td>G</td><td>G</td><td>G</td><td>G</td><td>G</td><td>H</td><td>G</td><td>D</td><td>G</td><td>D</td><td>G</td><td>H</td><td>G</td><td>D</td><td>G</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>201)</td>

<td>G</td><td>G</td><td>G</td><td>G</td><td>G</td><td>H</td><td>G</td><td>E</td><td>G</td><td>E</td><td>G</td><td>H</td><td>G</td><td>E</td><td>G</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>202)</td>

<td>V</td><td>V</td><td>V</td><td>V</td><td>V</td><td>R</td><td>V</td><td>D</td><td>V</td><td>R</td><td>V</td><td>D</td><td>V</td><td>R</td><td>V</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>203)</td>

<td>V</td><td>V</td><td>V</td><td>V</td><td>V</td><td>R</td><td>V</td><td>R</td><td>V</td><td>D</td><td>V</td><td>D</td><td>V</td><td>R</td><td>V</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>204)</td>

<td>V</td><td>V</td><td>V</td><td>V</td><td>V</td><td>E</td><td>V</td><td>K</td><td>V</td><td>E</td><td>V</td><td>K</td><td>V</td><td>E</td><td>V</td><td>K</td><td>(SEQ.</td><td>ID NO.</td><td>205)</td>

<td>V</td><td>V</td><td>V</td><td>V</td><td>V</td><td>E</td><td>V</td><td>E</td><td>V</td><td>K</td><td>V</td><td>K</td><td>V</td><td>E</td><td>V</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>206)</td>

<td>V</td><td>V</td><td>V</td><td>V</td><td>V</td><td>R</td><td>V</td><td>E</td><td>V</td><td>R</td><td>V</td><td>E</td><td>V</td><td>R</td><td>V</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>207)</td>

<td>V</td><td>V</td><td>V</td><td>V</td><td>V</td><td>R</td><td>V</td><td>R</td><td>V</td><td>E</td><td>V</td><td>E</td><td>V</td><td>R</td><td>V</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>208)</td>

<td>V</td><td>V</td><td>V</td><td>V</td><td>V</td><td>K</td><td>V</td><td>D</td><td>V</td><td>K</td><td>V</td><td>D</td><td>V</td><td>K</td><td>V</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>209)</td>

<td>V</td><td>V</td><td>V</td><td>V</td><td>V</td><td>E</td><td>V</td><td>H</td><td>V</td><td>E</td><td>V</td><td>H</td><td>V</td><td>E</td><td>V</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>210)</td>

<td>V</td><td>V</td><td>V</td><td>V</td><td>V</td><td>E</td><td>V</td><td>E</td><td>V</td><td>H</td><td>V</td><td>H</td><td>V</td><td>E</td><td>V</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>211)</td>

<td>V</td><td>V</td><td>V</td><td>V</td><td>V</td><td>R</td><td>V</td><td>R</td><td>V</td><td>R</td><td>V</td><td>R</td><td>V</td><td>R</td><td>V</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>212)</td>

<td>V</td><td>V</td><td>V</td><td>V</td><td>V</td><td>R</td><td>V</td><td>R</td><td>V</td><td>R</td><td>V</td><td>R</td><td>V</td><td>D</td><td>V</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>213)</td>

<td>V</td><td>V</td><td>V</td><td>V</td><td>V</td><td>R</td><td>V</td><td>R</td><td>V</td><td>R</td><td>V</td><td>D</td><td>V</td><td>D</td><td>V</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>214)</td>

<td>V</td><td>V</td><td>V</td><td>V</td><td>V</td><td>H</td><td>V</td><td>D</td><td>V</td><td>H</td><td>V</td><td>D</td><td>V</td><td>H</td><td>V</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>215)</td>

<td>V</td><td>V</td><td>V</td><td>V</td><td>V</td><td>H</td><td>V</td><td>H</td><td>V</td><td>H</td><td>V</td><td>H</td><td>V</td><td>H</td><td>V</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>216)</td>

<td>V</td><td>V</td><td>V</td><td>V</td><td>V</td><td>H</td><td>V</td><td>D</td><td>V</td><td>D</td><td>V</td><td>H</td><td>V</td><td>D</td><td>V</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>217)</td>

<td>V</td><td>V</td><td>V</td><td>V</td><td>V</td><td>H</td><td>V</td><td>E</td><td>V</td><td>E</td><td>V</td><td>H</td><td>V</td><td>E</td><td>V</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>218)</td>

<td>L</td><td>L</td><td>L</td><td>L</td><td>L</td><td>R</td><td>L</td><td>D</td><td>L</td><td>R</td><td>L</td><td>D</td><td>L</td><td>R</td><td>L</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>219)</td>

<td>95</td><td>L</td><td>L</td><td>L</td><td>L</td><td>L</td><td>R</td><td>L</td><td>R</td><td>L</td><td>D</td><td>L</td><td>D</td><td>L</td><td>R</td><td>L</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>220)</td>

<td>96</td><td>L</td><td>L</td><td>L</td><td>L</td><td>L</td><td>E</td><td>L</td><td>K</td><td>L</td><td>E</td><td>L</td><td>K</td><td>L</td><td>E</td><td>L</td><td>K</td><td>(SEQ.</td><td>ID NO.</td><td>221)</td>

<td>97</td><td>L</td><td>L</td><td>L</td><td>L</td><td>L</td><td>E</td><td>L</td><td>E</td><td>L</td><td>K</td><td>L</td><td>K</td><td>L</td><td>E</td><td>L</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>222)</td>

<td>98</td><td>L</td><td>L</td><td>L</td><td>L</td><td>L</td><td>R</td><td>L</td><td>E</td><td>L</td><td>R</td><td>L</td><td>E</td><td>L</td><td>R</td><td>L</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>223)</td>

<td>99</td><td>L</td><td>L</td><td>L</td><td>L</td><td>L</td><td>R</td><td>L</td><td>R</td><td>L</td><td>E</td><td>L</td><td>E</td><td>L</td><td>R</td><td>L</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>224)</td>

<td>100</td><td>L</td><td>L</td><td>L</td><td>L</td><td>L</td><td>K</td><td>L</td><td>D</td><td>L</td><td>K</td><td>L</td><td>D</td><td>L</td><td>K</td><td>L</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>225)</td>

<td>101</td><td>L</td><td>L</td><td>L</td><td>L</td><td>L</td><td>E</td><td>L</td><td>H</td><td>L</td><td>E</td><td>L</td><td>H</td><td>L</td><td>E</td><td>L</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>226)</td>

<td>102</td><td>L</td><td>L</td><td>L</td><td>L</td><td>L</td><td>E</td><td>L</td><td>E</td><td>L</td><td>H</td><td>L</td><td>H</td><td>L</td><td>E</td><td>L</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>227)</td>

<td>103</td><td>L</td><td>L</td><td>L</td><td>L</td><td>L</td><td>R</td><td>L</td><td>R</td><td>L</td><td>R</td><td>L</td><td>R</td><td>L</td><td>R</td><td>L</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>228)</td>

<td>104</td><td>L</td><td>L</td><td>L</td><td>L</td><td>L</td><td>R</td><td>L</td><td>R</td><td>L</td><td>R</td><td>L</td><td>R</td><td>L</td><td>D</td><td>L</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>229)</td>

<td>105</td><td>L</td><td>L</td><td>L</td><td>L</td><td>L</td><td>R</td><td>L</td><td>R</td><td>L</td><td>R</td><td>L</td><td>D</td><td>L</td><td>D</td><td>L</td><td>D</td><td colspan="3">(SEQ. ID NO.230)</td>

<td>106</td><td>L</td><td>L</td><td>L</td><td>L</td><td>L</td><td>H</td><td>L</td><td>D</td><td>L</td><td>H</td><td>L</td><td>D</td><td>L</td><td>H</td><td>L</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>231)</td>

<td>107</td><td>L</td><td>L</td><td>L</td><td>L</td><td>L</td><td>H</td><td>L</td><td>H</td><td>L</td><td>H</td><td>L</td><td>H</td><td>L</td><td>H</td><td>L</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>232)</td>

<td>108</td><td>L</td><td>L</td><td>L</td><td>L</td><td>L</td><td>H</td><td>L</td><td>D</td><td>L</td><td>D</td><td>L</td><td>H</td><td>L</td><td>D</td><td>L</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>233)</td>

<td>109</td><td>L</td><td>L</td><td>L</td><td>L</td><td>L</td><td>H</td><td>L</td><td>E</td><td>L</td><td>E</td><td>L</td><td>H</td><td>L</td><td>E</td><td>L</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>234)</td>

<td>110</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>R</td><td>AND</td><td>D</td><td>AND</td><td>R</td><td>AND</td><td>D</td><td>AND</td><td>R</td><td>AND</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>235)</td>

<td>111</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>D</td><td>AND</td><td>D</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>236)</td>

<td>112</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>E</td><td>AND</td><td>K</td><td>AND</td><td>E</td><td>AND</td><td>K</td><td>AND</td><td>E</td><td>AND</td><td>K</td><td>(SEQ.</td><td>ID NO.</td><td>237)</td>

<td>113</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>E</td><td>AND</td><td>E</td><td>AND</td><td>K</td><td>AND</td><td>K</td><td>AND</td><td>E</td><td>AND</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>238)</td>

<td>114</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>R</td><td>AND</td><td>E</td><td>AND</td><td>R</td><td>AND</td><td>E</td><td>AND</td><td>R</td><td>AND</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>239)</td>

<td>115</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>E</td><td>AND</td><td>E</td><td>AND</td><td>R</td><td>AND</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>240)</td>

<td>116</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>K</td><td>AND</td><td>D</td><td>AND</td><td>K</td><td>AND</td><td>D</td><td>AND</td><td>K</td><td>AND</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>241)</td>

<td>117</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>E</td><td>AND</td><td>H</td><td>AND</td><td>E</td><td>AND</td><td>H</td><td>AND</td><td>E</td><td>AND</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>242)</td>

<td>118</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>E</td><td>AND</td><td>E</td><td>AND</td><td>H</td><td>AND</td><td>H</td><td>AND</td><td>E</td><td>AND</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>243)</td>

<td>119</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>244)</td>

<td>120</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>D</td><td>AND</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>245)</td>

<td>121</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>R</td><td>AND</td><td>D</td><td>AND</td><td>D</td><td>AND</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>246)</td>

<td>122</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>H</td><td>AND</td><td>D</td><td>AND</td><td>H</td><td>AND</td><td>D</td><td>AND</td><td>H</td><td>AND</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>247)</td>

<td>123</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>H</td><td>AND</td><td>H</td><td>AND</td><td>H</td><td>AND</td><td>H</td><td>AND</td><td>H</td><td>AND</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>248)</td>

<td>124</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>H</td><td>AND</td><td>D</td><td>AND</td><td>D</td><td>AND</td><td>H</td><td>AND</td><td>D</td><td>AND</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>249)</td>

<td>125</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>AND</td><td>H</td><td>AND</td><td>E</td><td>AND</td><td>E</td><td>AND</td><td>H</td><td>AND</td><td>E</td><td>AND</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>250)</td>

<td>126</td><td>M</td><td>M</td><td>M</td><td>M</td><td>M</td><td>R</td><td>M</td><td>D</td><td>M</td><td>R</td><td>M</td><td>D</td><td>M</td><td>R</td><td>M</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>251)</td>

<td>127</td><td>M</td><td>M</td><td>M</td><td>M</td><td>M</td><td>R</td><td>M</td><td>R</td><td>M</td><td>D</td><td>M</td><td>D</td><td>M</td><td>R</td><td>M</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>252)</td>

<td>128</td><td>M</td><td>M</td><td>M</td><td>M</td><td>M</td><td>E</td><td>M</td><td>K</td><td>M</td><td>E</td><td>M</td><td>K</td><td>M</td><td>E</td><td>M</td><td>K</td><td>(SEQ.</td><td>ID NO.</td><td>253)</td>

<td>M</td><td>M</td><td>M</td><td>M</td><td>M</td><td>E</td><td>M</td><td>E</td><td>M</td><td>K</td><td>M</td><td>K</td><td>M</td><td>E</td><td>M</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>254)</td>

<td>M</td><td>M</td><td>M</td><td>M</td><td>M</td><td>R</td><td>M</td><td>E</td><td>M</td><td>R</td><td>M</td><td>E</td><td>M</td><td>R</td><td>M</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>255)</td>

<td>M</td><td>M</td><td>M</td><td>M</td><td>M</td><td>R</td><td>M</td><td>R</td><td>M</td><td>E</td><td>M</td><td>E</td><td>M</td><td>R</td><td>M</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>256)</td>

<td>M</td><td>M</td><td>M</td><td>M</td><td>M</td><td>K</td><td>M</td><td>D</td><td>M</td><td>K</td><td>M</td><td>D</td><td>M</td><td>K</td><td>M</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>257)</td>

<td>M</td><td>M</td><td>M</td><td>M</td><td>M</td><td>E</td><td>M</td><td>H</td><td>M</td><td>E</td><td>M</td><td>H</td><td>M</td><td>E</td><td>M</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>258)</td>

<td>M</td><td>M</td><td>M</td><td>M</td><td>M</td><td>E</td><td>M</td><td>E</td><td>M</td><td>H</td><td>M</td><td>H</td><td>M</td><td>E</td><td>M</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>259)</td>

<td>M</td><td>M</td><td>M</td><td>M</td><td>M</td><td>R</td><td>M</td><td>R</td><td>M</td><td>R</td><td>M</td><td>R</td><td>M</td><td>R</td><td>M</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>260)</td>

<td>M</td><td>M</td><td>M</td><td>M</td><td>M</td><td>R</td><td>M</td><td>R</td><td>M</td><td>R</td><td>M</td><td>R</td><td>M</td><td>D</td><td>M</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>261)</td>

<td>M</td><td>M</td><td>M</td><td>M</td><td>M</td><td>R</td><td>M</td><td>R</td><td>M</td><td>R</td><td>M</td><td>D</td><td>M</td><td>D</td><td>M</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>262)</td>

<td>M</td><td>M</td><td>M</td><td>M</td><td>M</td><td>H</td><td>M</td><td>D</td><td>M</td><td>H</td><td>M</td><td>D</td><td>M</td><td>H</td><td>M</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>263)</td>

<td>M</td><td>M</td><td>M</td><td>M</td><td>M</td><td>H</td><td>M</td><td>H</td><td>M</td><td>H</td><td>M</td><td>H</td><td>M</td><td>H</td><td>M</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>264)</td>

<td>M</td><td>M</td><td>M</td><td>M</td><td>M</td><td>H</td><td>M</td><td>D</td><td>M</td><td>D</td><td>M</td><td>H</td><td>M</td><td>D</td><td>M</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>265)</td>

<td>M</td><td>M</td><td>M</td><td>M</td><td>M</td><td>H</td><td>M</td><td>E</td><td>M</td><td>E</td><td>M</td><td>H</td><td>M</td><td>E</td><td>M</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>266)</td>

<td>F</td><td>F</td><td>F</td><td>F</td><td>F</td><td>R</td><td>F</td><td>D</td><td>F</td><td>R</td><td>F</td><td>D</td><td>F</td><td>R</td><td>F</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>267)</td>

<td>F</td><td>F</td><td>F</td><td>F</td><td>F</td><td>R</td><td>F</td><td>R</td><td>F</td><td>D</td><td>F</td><td>D</td><td>F</td><td>R</td><td>F</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>268)</td>

<td>F</td><td>F</td><td>F</td><td>F</td><td>F</td><td>E</td><td>F</td><td>K</td><td>F</td><td>E</td><td>F</td><td>K</td><td>F</td><td>E</td><td>F</td><td>K</td><td>(SEQ.</td><td>ID NO.</td><td>269)</td>

<td>F</td><td>F</td><td>F</td><td>F</td><td>F</td><td>E</td><td>F</td><td>E</td><td>F</td><td>K</td><td>F</td><td>K</td><td>F</td><td>E</td><td>F</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>270)</td>

<td>F</td><td>F</td><td>F</td><td>F</td><td>F</td><td>R</td><td>F</td><td>E</td><td>F</td><td>R</td><td>F</td><td>E</td><td>F</td><td>R</td><td>F</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>271)</td>

<td>F</td><td>F</td><td>F</td><td>F</td><td>F</td><td>R</td><td>F</td><td>R</td><td>F</td><td>E</td><td>F</td><td>E</td><td>F</td><td>R</td><td>F</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>272)</td>

<td>F</td><td>F</td><td>F</td><td>F</td><td>F</td><td>K</td><td>F</td><td>D</td><td>F</td><td>K</td><td>F</td><td>D</td><td>F</td><td>K</td><td>F</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>273)</td>

<td>F</td><td>F</td><td>F</td><td>F</td><td>F</td><td>E</td><td>F</td><td>H</td><td>F</td><td>E</td><td>F</td><td>H</td><td>F</td><td>E</td><td>F</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>274)</td>

<td>F</td><td>F</td><td>F</td><td>F</td><td>F</td><td>E</td><td>F</td><td>E</td><td>F</td><td>H</td><td>F</td><td>H</td><td>F</td><td>E</td><td>F</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>275)</td>

<td>F</td><td>F</td><td>F</td><td>F</td><td>F</td><td>R</td><td>F</td><td>R</td><td>F</td><td>R</td><td>F</td><td>R</td><td>F</td><td>R</td><td>F</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>276)</td>

<td>F</td><td>F</td><td>F</td><td>F</td><td>F</td><td>R</td><td>F</td><td>R</td><td>F</td><td>R</td><td>F</td><td>R</td><td>F</td><td>D</td><td>F</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>277)</td>

<td>F</td><td>F</td><td>F</td><td>F</td><td>F</td><td>R</td><td>F</td><td>R</td><td>F</td><td>R</td><td>F</td><td>D</td><td>F</td><td>D</td><td>F</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>27 8)</td>

<td>F</td><td>F</td><td>F</td><td>F</td><td>F</td><td>H</td><td>F</td><td>D</td><td>F</td><td>H</td><td>F</td><td>D</td><td>F</td><td>H</td><td>F</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>279)</td>

<td>F</td><td>F</td><td>F</td><td>F</td><td>F</td><td>H</td><td>F</td><td>H</td><td>F</td><td>H</td><td>F</td><td>H</td><td>F</td><td>H</td><td>F</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>280)</td>

<td>F</td><td>F</td><td>F</td><td>F</td><td>F</td><td>H</td><td>F</td><td>D</td><td>F</td><td>D</td><td>F</td><td>H</td><td>F</td><td>D</td><td>F</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>281)</td>

<td>F</td><td>F</td><td>F</td><td>F</td><td>F</td><td>H</td><td>F</td><td>E</td><td>F</td><td>E</td><td>F</td><td>H</td><td>F</td><td>E</td><td>F</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>282)</td>

<td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>R</td><td>IN</td><td>D</td><td>IN</td><td>R</td><td>IN</td><td>D</td><td>IN</td><td>R</td><td>IN</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>283)</td>

<td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>R</td><td>IN</td><td>R</td><td>IN</td><td>D</td><td>IN</td><td>D</td><td>IN</td><td>R</td><td>IN</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>284)</td>

<td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>E</td><td>IN</td><td>K</td><td>IN</td><td>E</td><td>IN</td><td>K</td><td>IN</td><td>E</td><td>IN</td><td>K</td><td>(SEQ.</td><td>ID NO.</td><td>285)</td>

<td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>E</td><td>IN</td><td>E</td><td>IN</td><td>K</td><td>IN</td><td>K</td><td>IN</td><td>E</td><td>IN</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>286)</td>

<td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>R</td><td>IN</td><td>E</td><td>IN</td><td>R</td><td>IN</td><td>E</td><td>IN</td><td>R</td><td>IN</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>287)</td>

<td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>R</td><td>IN</td><td>R</td><td>IN</td><td>E</td><td>IN</td><td>E</td><td>IN</td><td>R</td><td>IN</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>288)</td>

<td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>K</td><td>IN</td><td>D</td><td>IN</td><td>K</td><td>IN</td><td>D</td><td>IN</td><td>K</td><td>IN</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>289)</td>

<td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>E</td><td>IN</td><td>H</td><td>IN</td><td>E</td><td>IN</td><td>H</td><td>IN</td><td>E</td><td>IN</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>290)</td>

<td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>E</td><td>IN</td><td>E</td><td>IN</td><td>H</td><td>IN</td><td>H</td><td>IN</td><td>E</td><td>IN</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>291)</td>

<td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>R</td><td>IN</td><td>R</td><td>IN</td><td>R</td><td>IN</td><td>R</td><td>IN</td><td>R</td><td>IN</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>292)</td>

<td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>R</td><td>IN</td><td>R</td><td>IN</td><td>R</td><td>IN</td><td>R</td><td>IN</td><td>D</td><td>IN</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>293)</td>

<td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>R</td><td>IN</td><td>R</td><td>IN</td><td>R</td><td>IN</td><td>D</td><td>IN</td><td>D</td><td>IN</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>294)</td>

<td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>H</td><td>IN</td><td>D</td><td>IN</td><td>H</td><td>IN</td><td>D</td><td>IN</td><td>H</td><td>IN</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>295)</td>

<td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>H</td><td>IN</td><td>H</td><td>IN</td><td>H</td><td>IN</td><td>H</td><td>IN</td><td>H</td><td>IN</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>296)</td>

<td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>H</td><td>IN</td><td>D</td><td>IN</td><td>D</td><td>IN</td><td>H</td><td>IN</td><td>D</td><td>IN</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>297)</td>

<td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>IN</td><td>H</td><td>IN</td><td>E</td><td>IN</td><td>E</td><td>IN</td><td>H</td><td>IN</td><td>E</td><td>IN</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>298)</td>

<td>P</td><td>P</td><td>P</td><td>P</td><td>P</td><td>R</td><td>P</td><td>D</td><td>P</td><td>R</td><td>P</td><td>D</td><td>P</td><td>R</td><td>P</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>299)</td>

<td>P</td><td>P</td><td>P</td><td>P</td><td>P</td><td>R</td><td>P</td><td>R</td><td>P</td><td>D</td><td>P</td><td>D</td><td>P</td><td>R</td><td>P</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>300)</td>

<td>P</td><td>P</td><td>P</td><td>P</td><td>P</td><td>E</td><td>P</td><td>K</td><td>P</td><td>E</td><td>P</td><td>K</td><td>P</td><td>E</td><td>P</td><td>K</td><td>(SEQ.</td><td>ID NO.</td><td>301)</td>

<td>P</td><td>P</td><td>P</td><td>P</td><td>P</td><td>E</td><td>P</td><td>E</td><td>P</td><td>K</td><td>P</td><td>K</td><td>P</td><td>E</td><td>P</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>302)</td>

<td>P</td><td>P</td><td>P</td><td>P</td><td>P</td><td>R</td><td>P</td><td>E</td><td>P</td><td>R</td><td>P</td><td>E</td><td>P</td><td>R</td><td>P</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>303)</td>

<td>P</td><td>P</td><td>P</td><td>P</td><td>P</td><td>R</td><td>P</td><td>R</td><td>P</td><td>E</td><td>P</td><td>E</td><td>P</td><td>R</td><td>P</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>304)</td>

<td>P</td><td>P</td><td>P</td><td>P</td><td>P</td><td>K</td><td>P</td><td>D</td><td>P</td><td>K</td><td>P</td><td>D</td><td>P</td><td>K</td><td>P</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>305)</td>

<td>P</td><td>P</td><td>P</td><td>P</td><td>P</td><td>E</td><td>P</td><td>H</td><td>P</td><td>E</td><td>P</td><td>H</td><td>P</td><td>E</td><td>P</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>306)</td>

<td>P</td><td>P</td><td>P</td><td>P</td><td>P</td><td>E</td><td>P</td><td>E</td><td>P</td><td>H</td><td>P</td><td>H</td><td>P</td><td>E</td><td>P</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>307)</td>

<td>P</td><td>P</td><td>P</td><td>P</td><td>P</td><td>R</td><td>P</td><td>R</td><td>P</td><td>R</td><td>P</td><td>R</td><td>P</td><td>R</td><td>P</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>308)</td>

<td>P</td><td>P</td><td>P</td><td>P</td><td>P</td><td>R</td><td>P</td><td>R</td><td>P</td><td>R</td><td>P</td><td>R</td><td>P</td><td>D</td><td>P</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>309)</td>

<td>P</td><td>P</td><td>P</td><td>P</td><td>P</td><td>R</td><td>P</td><td>R</td><td>P</td><td>R</td><td>P</td><td>D</td><td>P</td><td>D</td><td>P</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>310)</td>

<td>P</td><td>P</td><td>P</td><td>P</td><td>P</td><td>H</td><td>P</td><td>D</td><td>P</td><td>H</td><td>P</td><td>D</td><td>P</td><td>H</td><td>P</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>311)</td>

<td>P</td><td>P</td><td>P</td><td>P</td><td>P</td><td>H</td><td>P</td><td>H</td><td>P</td><td>H</td><td>P</td><td>H</td><td>P</td><td>H</td><td>P</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>312)</td>

<td>P</td><td>P</td><td>P</td><td>P</td><td>P</td><td>H</td><td>P</td><td>D</td><td>P</td><td>D</td><td>P</td><td>H</td><td>P</td><td>D</td><td>P</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>313)</td>

<td>P</td><td>P</td><td>P</td><td>P</td><td>P</td><td>H</td><td>P</td><td>E</td><td>P</td><td>E</td><td>P</td><td>H</td><td>P</td><td>E</td><td>P</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>314)</td>

<td>S</td><td>S</td><td>S</td><td>S</td><td>S</td><td>R</td><td>S</td><td>D</td><td>S</td><td>R</td><td>S</td><td>D</td><td>S</td><td>R</td><td>S</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>315)</td>

<td>S</td><td>S</td><td>S</td><td>S</td><td>S</td><td>R</td><td>S</td><td>R</td><td>S</td><td>D</td><td>S</td><td>D</td><td>S</td><td>R</td><td>S</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>316)</td>

<td>S</td><td>S</td><td>S</td><td>S</td><td>S</td><td>E</td><td>S</td><td>K</td><td>S</td><td>E</td><td>S</td><td>K</td><td>S</td><td>E</td><td>S</td><td>K</td><td>(SEQ.</td><td>ID NO.</td><td>317)</td>

<td>S</td><td>S</td><td>S</td><td>S</td><td>S</td><td>E</td><td>S</td><td>E</td><td>S</td><td>K</td><td>S</td><td>K</td><td>S</td><td>E</td><td>S</td><td>E</td><td colspan="3">(SEQ. ID NO.318)</td>

<td>S</td><td>S</td><td>S</td><td>S</td><td>S</td><td>R</td><td>S</td><td>E</td><td>S</td><td>R</td><td>S</td><td>E</td><td>S</td><td>R</td><td>S</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>319)</td>

<td>S</td><td>S</td><td>S</td><td>S</td><td>S</td><td>R</td><td>S</td><td>R</td><td>S</td><td>E</td><td>S</td><td>E</td><td>S</td><td>R</td><td>S</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>320)</td>

<td>S</td><td>S</td><td>S</td><td>S</td><td>S</td><td>K</td><td>S</td><td>D</td><td>S</td><td>K</td><td>S</td><td>D</td><td>S</td><td>K</td><td>S</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>321)</td>

<td>197</td><td>S</td><td>S</td><td>S</td><td>S</td><td>S</td><td>E</td><td>S</td><td>H</td><td>S</td><td>E</td><td>S</td><td>H</td><td>S</td><td>E</td><td>S</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>322)</td>

<td>198</td><td>S</td><td>S</td><td>S</td><td>S</td><td>S</td><td>E</td><td>S</td><td>E</td><td>S</td><td>H</td><td>S</td><td>H</td><td>S</td><td>E</td><td>S</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>323)</td>

<td>199</td><td>S</td><td>S</td><td>S</td><td>S</td><td>S</td><td>R</td><td>S</td><td>R</td><td>S</td><td>R</td><td>S</td><td>R</td><td>S</td><td>R</td><td>S</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>324)</td>

<td>200</td><td>S</td><td>S</td><td>S</td><td>S</td><td>S</td><td>R</td><td>S</td><td>R</td><td>S</td><td>R</td><td>S</td><td>R</td><td>S</td><td>D</td><td>S</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>325)</td>

<td>201</td><td>S</td><td>S</td><td>S</td><td>S</td><td>S</td><td>R</td><td>S</td><td>R</td><td>S</td><td>R</td><td>S</td><td>D</td><td>S</td><td>D</td><td>S</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>326)</td>

<td>202</td><td>S</td><td>S</td><td>S</td><td>S</td><td>S</td><td>H</td><td>S</td><td>D</td><td>S</td><td>H</td><td>S</td><td>D</td><td>S</td><td>H</td><td>S</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>327)</td>

<td>203</td><td>S</td><td>S</td><td>S</td><td>S</td><td>S</td><td>H</td><td>S</td><td>H</td><td>S</td><td>H</td><td>S</td><td>H</td><td>S</td><td>H</td><td>S</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>328)</td>

<td>204</td><td>S</td><td>S</td><td>S</td><td>S</td><td>S</td><td>H</td><td>S</td><td>D</td><td>S</td><td>D</td><td>S</td><td>H</td><td>S</td><td>D</td><td>S</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>329)</td>

<td>205</td><td>S</td><td>S</td><td>S</td><td>S</td><td>S</td><td>H</td><td>S</td><td>E</td><td>S</td><td>E</td><td>S</td><td>H</td><td>S</td><td>E</td><td>S</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>330)</td>

<td>206</td><td>T</td><td>T</td><td>T</td><td>T</td><td>T</td><td>R</td><td>T</td><td>D</td><td>T</td><td>R</td><td>T</td><td>D</td><td>T</td><td>R</td><td>T</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>331)</td>

<td>207</td><td>T</td><td>T</td><td>T</td><td>T</td><td>T</td><td>R</td><td>T</td><td>R</td><td>T</td><td>D</td><td>T</td><td>D</td><td>T</td><td>R</td><td>T</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>332)</td>

<td>208</td><td>T</td><td>T</td><td>T</td><td>T</td><td>T</td><td>E</td><td>T</td><td>K</td><td>T</td><td>E</td><td>T</td><td>K</td><td>T</td><td>E</td><td>T</td><td>K</td><td>(SEQ.</td><td>ID NO.</td><td>333)</td>

<td>209</td><td>T</td><td>T</td><td>T</td><td>T</td><td>T</td><td>E</td><td>T</td><td>E</td><td>T</td><td>K</td><td>T</td><td>K</td><td>T</td><td>E</td><td>T</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>334)</td>

<td>210</td><td>T</td><td>T</td><td>T</td><td>T</td><td>T</td><td>R</td><td>T</td><td>E</td><td>T</td><td>R</td><td>T</td><td>E</td><td>T</td><td>R</td><td>T</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>335)</td>

<td>211</td><td>T</td><td>T</td><td>T</td><td>T</td><td>T</td><td>R</td><td>T</td><td>R</td><td>T</td><td>E</td><td>T</td><td>E</td><td>T</td><td>R</td><td>T</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>336)</td>

<td>212</td><td>T</td><td>T</td><td>T</td><td>T</td><td>T</td><td>K</td><td>T</td><td>D</td><td>T</td><td>K</td><td>T</td><td>D</td><td>T</td><td>K</td><td>T</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>337)</td>

<td>213</td><td>T</td><td>T</td><td>T</td><td>T</td><td>T</td><td>E</td><td>T</td><td>H</td><td>T</td><td>E</td><td>T</td><td>H</td><td>T</td><td>E</td><td>T</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>338)</td>

<td>214</td><td>T</td><td>T</td><td>T</td><td>T</td><td>T</td><td>E</td><td>T</td><td>E</td><td>T</td><td>H</td><td>T</td><td>H</td><td>T</td><td>E</td><td>T</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>339)</td>

<td>215</td><td>T</td><td>T</td><td>T</td><td>T</td><td>T</td><td>R</td><td>T</td><td>R</td><td>T</td><td>R</td><td>T</td><td>R</td><td>T</td><td>R</td><td>T</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>340)</td>

<td>216</td><td>T</td><td>T</td><td>T</td><td>T</td><td>T</td><td>R</td><td>T</td><td>R</td><td>T</td><td>R</td><td>T</td><td>R</td><td>T</td><td>D</td><td>T</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>341)</td>

<td>217</td><td>T</td><td>T</td><td>T</td><td>T</td><td>T</td><td>R</td><td>T</td><td>R</td><td>T</td><td>R</td><td>T</td><td>D</td><td>T</td><td>D</td><td>T</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>342)</td>

<td>218</td><td>T</td><td>T</td><td>T</td><td>T</td><td>T</td><td>H</td><td>T</td><td>D</td><td>T</td><td>H</td><td>T</td><td>D</td><td>T</td><td>H</td><td>T</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>343)</td>

<td>219</td><td>T</td><td>T</td><td>T</td><td>T</td><td>T</td><td>H</td><td>T</td><td>H</td><td>T</td><td>H</td><td>T</td><td>H</td><td>T</td><td>H</td><td>T</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>344)</td>

<td>220</td><td>T</td><td>T</td><td>T</td><td>T</td><td>T</td><td>H</td><td>T</td><td>D</td><td>T</td><td>D</td><td>T</td><td>H</td><td>T</td><td>D</td><td>T</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>345)</td>

<td>221</td><td>T</td><td>T</td><td>T</td><td>T</td><td>T</td><td>H</td><td>T</td><td>E</td><td>T</td><td>E</td><td>T</td><td>H</td><td>T</td><td>E</td><td>T</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>346)</td>

<td>222</td><td>C</td><td>C</td><td>C</td><td>C</td><td>C</td><td>R</td><td>C</td><td>D</td><td>C</td><td>R</td><td>C</td><td>D</td><td>C</td><td>R</td><td>C</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>347)</td>

<td>223</td><td>C</td><td>C</td><td>C</td><td>C</td><td>C</td><td>R</td><td>C</td><td>R</td><td>C</td><td>D</td><td>C</td><td>D</td><td>C</td><td>R</td><td>C</td><td>R</td><td>(SEQ.</td><td>ID NO.</td><td>348)</td>

<td>224</td><td>C</td><td>C</td><td>C</td><td>C</td><td>C</td><td>E</td><td>C</td><td>K</td><td>C</td><td>E</td><td>C</td><td>K</td><td>C</td><td>E</td><td>C</td><td>K</td><td>(SEQ.</td><td>ID NO.</td><td>349)</td>

<td>225</td><td>C</td><td>C</td><td>C</td><td>C</td><td>C</td><td>E</td><td>C</td><td>E</td><td>C</td><td>K</td><td>C</td><td>K</td><td>C</td><td>E</td><td>C</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>350)</td>

<td>226</td><td>C</td><td>C</td><td>C</td><td>C</td><td>C</td><td>R</td><td>C</td><td>E</td><td>C</td><td>R</td><td>C</td><td>E</td><td>C</td><td>R</td><td>C</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>351)</td>

<td>227</td><td>C</td><td>C</td><td>C</td><td>C</td><td>C</td><td>R</td><td>C</td><td>R</td><td>C</td><td>E</td><td>C</td><td>E</td><td>C</td><td>R</td><td>C</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>352)</td>

<td>228</td><td>C</td><td>C</td><td>C</td><td>C</td><td>C</td><td>K</td><td>C</td><td>D</td><td>C</td><td>K</td><td>C</td><td>D</td><td>C</td><td>K</td><td>C</td><td>D</td><td>(SEQ.</td><td>ID NO.</td><td>353)</td>

<td>229</td><td>C</td><td>C</td><td>C</td><td>C</td><td>C</td><td>E</td><td>C</td><td>H</td><td>C</td><td>E</td><td>C</td><td>H</td><td>C</td><td>E</td><td>C</td><td>H</td><td>(SEQ.</td><td>ID NO.</td><td>354)</td>

<td>230</td><td>C</td><td>C</td><td>C</td><td>C</td><td>C</td><td>E</td><td>C</td><td>E</td><td>C</td><td>H</td><td>C</td><td>H</td><td>C</td><td>E</td><td>C</td><td>E</td><td>(SEQ.</td><td>ID NO.</td><td>355)</td>

<td>231</td><td>C</td><td>C</td><td>C</td><td>C</td><td>CRCRCRC</td><td>RCRCR (SEQ ID NO. 356)</td>

<td>232</td><td>C</td><td>C</td><td>C</td><td>C</td><td>CRCRCRC</td><td>RCDCD (SEQ ID NO. 357)</td>

<td>233</td><td>C</td><td>C</td><td>C</td><td>C</td><td>CRCRCRC</td><td>DCDCD (SEQ ID NO. 358)</td>

<td>234</td><td>C</td><td>C</td><td>C</td><td>C</td><td>CHCDCHC</td><td>DCHCD (SEQ ID NO: 359)</td>

<td>235</td><td>C</td><td>C</td><td>C</td><td>C</td><td>CHCHCHC</td><td>HCHCH (SEQ ID No. 360)</td>

<td>236</td><td>C</td><td>C</td><td>C</td><td>C</td><td>CHCDCDC</td><td>HCDCD (SEQ ID No. 361)</td>

<td>237</td><td>C</td><td>C</td><td>C</td><td>C</td><td>CHCECEC</td><td>HCECE (SEQ ID NO. 362)</td>

<td>238</td><td>Y</td><td>Y</td><td>Y</td><td>Y</td><td>YRYDYRY</td><td>DYRYD (SEQ ID NO: 363)</td>

<td>239</td><td>Y</td><td>Y</td><td>Y</td><td>Y</td><td>YRYRYDY</td><td>DYRYR (SEQ ID No. NO. 64)</td>

<td>240</td><td>Y</td><td>Y</td><td>Y</td><td>Y</td><td>YEYKYEY</td><td>KYEYK (SEQ ID NO. 365)</td>

<td>241</td><td>Y</td><td>Y</td><td>Y</td><td>Y</td><td>YEYEYKY</td><td>KYEYE (SEQ ID No. 366)</td>

<td>242</td><td>Y</td><td>Y</td><td>Y</td><td>Y</td><td>YRYEYRY</td><td>EYRYE (SEQ ID NO: 367)</td>

<td>243</td><td>Y</td><td>Y</td><td>Y</td><td>Y</td><td>YRYRYEY</td><td>EYRYE (SEQ ID NO.368)</td>

<td>244</td><td>Y</td><td>Y</td><td>Y</td><td>Y</td><td>YKYDYKY</td><td>DYKYD (SEQ ID No. 125)</td>

<td>245</td><td>Y</td><td>Y</td><td>Y</td><td>Y</td><td>YEYHYEY</td><td>HYEYH (SEQ ID NO. 369)</td>

<td>246</td><td>Y</td><td>Y</td><td>Y</td><td>Y</td><td>YEYEYHY</td><td>HYEYE (SEQ ID NO. 370)</td>

<td>247</td><td>Y</td><td>Y</td><td>Y</td><td>Y</td><td>YRYRYRY</td><td>RYRYR (SEQ ID NO: 371)</td>

<td>248</td><td>Y</td><td>Y</td><td>Y</td><td>Y</td><td>YRYRYRY</td><td>RYDYD (SEQ ID NO: 372)</td>

<td>249</td><td>Y</td><td>Y</td><td>Y</td><td>Y</td><td>YRYRYRY</td><td>DIDID (SEQ ID NO. 373)</td>

<td>250</td><td>Y</td><td>Y</td><td>Y</td><td>Y</td><td>YHYDYHY</td><td>DYHYD (SEQ ID No. 374)</td>

<td>251</td><td>Y</td><td>Y</td><td>Y</td><td>Y</td><td>YHYHYHY</td><td>HYHYH (SEQ ID NO. 375)</td>

<td>252</td><td>Y</td><td>Y</td><td>Y</td><td>Y</td><td>YHYDYDY</td><td>HYDYD (SEQ ID NO. 376)</td>

<td>253</td><td>Y</td><td>Y</td><td>Y</td><td>Y</td><td>YHYEYEY</td><td>HYEYE (SEQ ID NO: 377)</td>

<td>254</td><td>N</td><td>N</td><td>N</td><td>N</td><td>NRNDNRN</td><td>DNRND (SEQ ID NO: 378)</td>

<td>255</td><td>N</td><td>N</td><td>N</td><td>N</td><td>NRNRNDN</td><td>DNRNR (SEQ ID NO: 378)</td>

<td>256</td><td>N</td><td>N</td><td>N</td><td>N</td><td>NENKNEN</td><td>KNENK (SEQ ID NO 380)</td>

<td>257</td><td>N</td><td>N</td><td>N</td><td>N</td><td>NENENKN</td><td>KNENE (SEQ ID NO 381)</td>

<td>258</td><td>N</td><td>N</td><td>N</td><td>N</td><td>NRNENRN</td><td>ENRNE (SEQ ID NO 382)</td>

<td>259</td><td>N</td><td>N</td><td>N</td><td>N</td><td>NRNRNEN</td><td>ENRNE (SEQ ID NO 383)</td>

<td>260</td><td>N</td><td>N</td><td>N</td><td>N</td><td>NKNDNKN</td><td>DNKND (SEQ ID NO: 384)</td>

<td>261</td><td>N</td><td>N</td><td>N</td><td>N</td><td>NENHNEN</td><td>HNENH (SEQ ID NO: 385)</td>

<td>262</td><td>N</td><td>N</td><td>N</td><td>N</td><td>NENENHN</td><td>HNENE (SEQ ID NO 386)</td>

<td>263</td><td>N</td><td>N</td><td>N</td><td>N</td><td>NRNRNRN</td><td>RNRNR (SEQ ID NO 387)</td>

<td>264</td><td>N</td><td>N</td><td>N</td><td>N</td><td>NRNRNRN</td><td>RNDND (SEQ ID NO 388)</td>

<td>265</td><td>N</td><td>N</td><td>N</td><td>N</td><td>N</td><td>R</td><td>N</td><td>R</td><td>N</td><td>R</td><td>N</td><td>D</td><td>N</td><td>D</td><td>N</td><td>D</td><td>(SEQ ID NO.</td><td>389)</td>

<td>266</td><td>N</td><td>N</td><td>N</td><td>N</td><td>N</td><td>H</td><td>N</td><td>D</td><td>N</td><td>H</td><td>N</td><td>D</td><td>N</td><td>H</td><td>N</td><td>D</td><td>(SEQ ID NO.</td><td>390)</td>

<td>267</td><td>N</td><td>N</td><td>N</td><td>N</td><td>N</td><td>H</td><td>N</td><td>H</td><td>N</td><td>H</td><td>N</td><td>H</td><td>N</td><td>H</td><td>N</td><td>H</td><td>(SEQ ID NO.</td><td>391)</td>

<td>268</td><td>N</td><td>N</td><td>N</td><td>N</td><td>N</td><td>H</td><td>N</td><td>D</td><td>N</td><td>D</td><td>N</td><td>H</td><td>N</td><td>D</td><td>N</td><td>D</td><td>(SEQ ID NO.</td><td>392)</td>

<td>269</td><td>N</td><td>N</td><td>N</td><td>N</td><td>N</td><td>H</td><td>N</td><td>E</td><td>N</td><td>E</td><td>N</td><td>H</td><td>N</td><td>E</td><td>N</td><td>E</td><td>(SEQ ID NO.</td><td>393)</td>

<td>270</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>R</td><td>^Q</td><td>D</td><td>^Q</td><td>R</td><td>^Q</td><td>D</td><td>^Q</td><td>R</td><td>^Q</td><td>D</td><td>(SEQ ID NO.</td><td>394)</td>

<td>271</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>R</td><td>^Q</td><td>R</td><td>^Q</td><td>D</td><td>^Q</td><td>D</td><td>^Q</td><td>R</td><td>^Q</td><td>R</td><td>(SEQ ID NO.</td><td>395)</td>

<td>272</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>E</td><td>^Q</td><td>K</td><td>^Q</td><td>E</td><td>^Q</td><td>K</td><td>^Q</td><td>E</td><td>^Q</td><td>K</td><td colspan="2">(SEQ. ID NO.396)</td>

<td>273</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>E</td><td>^Q</td><td>E</td><td>^Q</td><td>K</td><td>^Q</td><td>K</td><td>^Q</td><td>E</td><td>^Q</td><td>E</td><td>(SEQ ID NO.</td><td>397)</td>

<td>274</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>R</td><td>^Q</td><td>E</td><td>^Q</td><td>R</td><td>^Q</td><td>E</td><td>^Q</td><td>R</td><td>^Q</td><td>E</td><td>(SEQ ID NO.</td><td>398)</td>

<td>275</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>R</td><td>^Q</td><td>R</td><td>^Q</td><td>E</td><td>^Q</td><td>E</td><td>^Q</td><td>R</td><td>^Q</td><td>E</td><td>(SEQ ID NO.</td><td>399)</td>

<td>276</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>K</td><td>^Q</td><td>D</td><td>^Q</td><td>K</td><td>^Q</td><td>D</td><td>^Q</td><td>K</td><td>^Q</td><td>D</td><td>(SEQ ID NO.</td><td>400)</td>

<td>277</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>E</td><td>^Q</td><td>H</td><td>^Q</td><td>E</td><td>^Q</td><td>H</td><td>^Q</td><td>E</td><td>^Q</td><td>H</td><td>(SEQ ID NO.</td><td>401)</td>

<td>278</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>E</td><td>^Q</td><td>E</td><td>^Q</td><td>H</td><td>^Q</td><td>H</td><td>^Q</td><td>E</td><td>^Q</td><td>E</td><td>(SEQ ID NO.</td><td>402)</td>

<td>279</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>R</td><td>^Q</td><td>R</td><td>^Q</td><td>R</td><td>^Q</td><td>R</td><td>^Q</td><td>R</td><td>^Q</td><td>R</td><td>(SEQ ID NO.</td><td>403)</td>

<td>280</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>R</td><td>^Q</td><td>R</td><td>^Q</td><td>R</td><td>^Q</td><td>R</td><td>^Q</td><td>D</td><td>^Q</td><td>D</td><td>(SEQ ID NO.</td><td>404)</td>

<td>281</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>R</td><td>^Q</td><td>R</td><td>^Q</td><td>R</td><td>^Q</td><td>D</td><td>^Q</td><td>D</td><td>^Q</td><td>D</td><td>(SEQ ID NO.</td><td>405)</td>

<td>282</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>H</td><td>^Q</td><td>D</td><td>^Q</td><td>H</td><td>^Q</td><td>D</td><td>^Q</td><td>H</td><td>^Q</td><td>D</td><td>(SEQ ID NO.</td><td>406)</td>

<td>283</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>H</td><td>^Q</td><td>H</td><td>^Q</td><td>H</td><td>^Q</td><td>H</td><td>^Q</td><td>H</td><td>^Q</td><td>H</td><td>(SEQ ID NO.</td><td>407)</td>

<td>284</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>H</td><td>^Q</td><td>D</td><td>^Q</td><td>D</td><td>^Q</td><td>H</td><td>^Q</td><td>D</td><td>^Q</td><td>D</td><td>(SEQ ID NO.</td><td>408)</td>

<td>285</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>^Q</td><td>H</td><td>^Q</td><td>E</td><td>^Q</td><td>E</td><td>^Q</td><td>H</td><td>^Q</td><td>E</td><td>^Q</td><td>E</td><td>(SEQ ID NO.</td><td>409)</td>

[0079] Self-assembling peptides are typically linear sequences. However, the self-assembling peptides may be in the form of non-linear sequences, optionally including hydrophobic tails that interact with the ECM. In one embodiment, the sequence is in the form of a "rake" in which the ridge teeth are hydrophobic sequences that interact with the ECM to anchor the self-assembled peptides in a tissue or vessel. The rake holder includes a sequence that self-assembles. In another embodiment, the rake holder is a hydrophobic sequence that interacts with the ECM, and the rake teeth are sequences that self-assemble. The self-assembling sequences may self-assemble or in the presence of one or more assembly-assisting sequences.

D. Therapeutic, prophylactic and diagnostic agents [0080] The formulations may also contain other therapeutic, prophylactic or diagnostic agents. In a preferred embodiment, these may be anti-inflammatory agents, vasoconstrictors, anti-infective agents, anesthetics, growth factors, vitamins, nutrients and / or cells.

[0081] These may be peptides or proteins, polysaccharides or saccharides, nucleic acid nucleic acids, a proteoglycan, lipid, carbohydrate or small molecule, usually an organic compound having multiple carbon-carbon bonds that may be isolated from nature or produced by chemical synthesis. Small molecules have relatively small molecular weights (e.g., less than about 1500 g / mol) and are not peptides or nucleic acids. The substance may also be a biomolecule, which is a molecule such as a peptide, a proteoglycan, a lipid, a carbohydrate or a nucleic acid, exhibiting characteristics typical of molecules found in living organisms. Like small molecules, biomolecules may be naturally occurring or they may be artificial (i.e. they may be molecules that are not found in nature). For example, a protein having a sequence, which is not present in the family (e.g., does not exist in a publicly accessible sequence database), or which includes a known sequence modified in an unnatural manner by a human (e.g., a sequence modified by a post-translational processing such as glycosylation) is an artificial biomolecule. Nucleic acid molecules encoding such proteins (e.g., an oligonucleotide, optionally contained in an expression vector) are also biomolecules and can be included in the compositions described herein. For example, a composition may contain a plurality of self-assembled materials and cells that express or are designed to express a biomolecule of a protein (due to the inclusion of a nucleic acid sequence that encodes a protein biomolecule). or which includes a known sequence modified in an unnatural manner by a human (e.g., a sequence modified by a post-translational modification such as glycosylation) is an artificial biomolecule. Nucleic acid molecules encoding such proteins (e.g., an oligonucleotide, optionally contained in an expression vector) are also biomolecules and can be included in the compositions described herein. For example, a composition may contain a plurality of self-assembled materials and cells that express or are designed to express a biomolecule of a protein (due to the inclusion of a nucleic acid sequence that encodes a protein biomolecule). or which includes a known sequence modified in an unnatural manner by a human (e.g., a sequence modified by a post-translational modification such as glycosylation) is an artificial biomolecule. Nucleic acid molecules encoding such proteins (e.g., an oligonucleotide, optionally contained in an expression vector) are also biomolecules and can be included in the compositions described herein. For example, a composition may contain a plurality of self-assembled materials and cells that express or are designed to express a biomolecule of a protein (due to the inclusion of a nucleic acid sequence that encodes a protein biomolecule). Nucleic acid molecules encoding such proteins (e.g., an oligonucleotide, optionally contained in an expression vector) are also biomolecules and can be included in the compositions described herein. For example, a composition may contain a plurality of self-assembled materials and cells that express or are designed to express a biomolecule of a protein (due to the inclusion of a nucleic acid sequence that encodes a protein biomolecule). Nucleic acid molecules encoding such proteins (e.g., an oligonucleotide, optionally contained in an expression vector) are also biomolecules and can be included in the compositions described herein. For example, a composition may contain a plurality of self-assembled materials and cells that express or are designed to express a biomolecule of a protein (due to the inclusion of a nucleic acid sequence that encodes a protein biomolecule).

[0082] A variety of therapeutic, prophylactic or diagnostic agents may be included in the formulation. Representative vasoconstrictors include epinephrine and phenylephrine; representative colorants include arsenazo III, chlorophosphonazo III, antipyrylazo 111, murexide, eriochromic black T, eriochrome blue SE, oxyacetazo I, carboxyazo III, tropolone, methylthymol blue and black Mordant 32; representative anesthetics include benzocaine, bupivacaine, bubbumin picrate, chloroprocaine, cocaine, curare, dibucin, diclonine, etidocaine, lidocaine, mepivacaine, pramoxin, prilocaine, procaine, propoxicin, ropivacaine, tetracaine or combinations thereof. Local use of an anesthetic may be all that is required in some situations, e.g. in the event of a burn or other skin wound, including pressure ulcers; wounds, such as cancer ulcers; or for minimally invasive surgery. Combining local anesthetics with self-assembling peptides, whether in combination with their presence in the same preparation or simultaneous administration, can help stop the anesthetic in the body and reduce the amount entering the circulation.

[0083] Vasoconstrictors such as phenylephrine may be introduced to prolong the action of local general anesthesia (e.g., 0.1-0.5% phenylephrine). Analgesics other than a local anesthetic, such as steroids, non-steroidal anti-inflammatory agents, such as indomethacin, platelet activating factor (PAF) inhibitors such as lexipaphant, CV 3988 and / or PAF receptor inhibitors such as SRI 63441.

[0084] For systemic or local administration, an anti-infective or anti-microbial agent (e.g., antibiotic, anti-bacterial, antiviral or antifungal) may be included. Examples include β-lactam antibiotics, such as polylactins and cephalosporins; other cell wall synthesis inhibitors such as vancomycin; chloramphenicol; tetracycline; macrolides; clindamycin; streptogramins; aminoglycosides; spectinomycin; sulfonamides; trimethoprim; quinolones; amphotericin B; flucytosine; azoles, such as ketoconazole, itraconazole, fluconazole, clotrimazole and mikonazol; griseofulvin; terbinafine; and nystatin. Antimicrobials can be administered topically (e.g., in the treatment of skin infections or burns) or to help prevent infection at the catheter insertion site (e.g., an intravenous catheter). Suitable topical antimicrobials include kanamycin, neomycin, bacitracin, polymyxin, topical sulfonamides such as mafenid acetate or silver sulfodiazine and gentamycin sulfate. The antimicrobial agent can also be a broad-spectrum agent. For example, a second, third or fourth generation cephalosporin may be used. These factors may be active against a wide range of bacteria, including Gram-positive and Gram-negative species. Such antibacterial agents may be particularly suitable if the present scaffolds are used to inhibit the flow of intestinal contents, such as during intestinal resection or other surgical procedures that deliberately or accidentally interfere with intestinal wall integrity. One of ordinary skill in the art will be able to choose appropriate antibacterial agents, taking into account such factors as the patient's history (eg the history of an allergic reaction to such factors), the place where the peptides are to be used, and the type of infectious agent that may be present. Compositions containing antibacterial agents can prevent infections in various ways, including: (1) killing an infectious agent due to the action of an antibacterial; (2) preventing infection by assembling peptides to form a barrier that blocks the infiltration of the infectious agent into the tissue by blocking the interaction with the tissue of the tissue-specific sequence of an infectious agent; (3) causing the infectious agent to change the orientation relative to the tissue due to the charge of the self-depositing material, thereby blocking the infiltration of the infectious agent into the tissue; (4) encapsulation of an infectious agent within self-assembling peptides to prevent infection of the infectious agent; and their connections. Self-assembling peptides can also be used to prevent contamination or infection with other biological and / or hazardous materials.

[0085] Each of the compositions described herein, regardless of whether it contains only self-assembling peptides and one or more bioactive molecules (and whether in liquid, semi-solid or solid form), may contain a coloring agent. Suitable colorants include commercially available food colors, natural and synthetic dyes, and fluorescent molecules. Preferably, the colorant is nontoxic or is included at low concentrations to minimize any toxic effects. The use of a coloring agent allows a better visualization of the area that is covered by the structure or scaffolding and can facilitate removal if such removal is desired. The colorant may change color in contact with the contaminated area (e.g., color change may be caused by the same contamination (e.g. by blood or bacteria present at the site of the wound)). For example, a metabolic product of bacteria can cause a change in color. It is also possible to detect conditions such as the pH or redox state induced by contaminating agents. Exemplary indicators include arsenzase III, chlorophosphonazo III, antipyrylazo III, murexide, epichromic black T and epidermal blue SE for Mg²⁺oxyacetazo I, carboxyazo III, tropolone, methylthymol blue and black Mordant 32. AlamarBlue, redox index and phenol red also have applications in compositions and methods. In another embodiment, the colorant may be in the form of a nanoparticle that reflects one wavelength of light, and after aggregation (i.e., self-assembly of the peptide) reflects a different wavelength of light.

[0086] A variety of other active agents may be included in the composition. For example, to accelerate one or more aspects of healing (e.g., angiogenesis, cell migration, process prolongation and cell proliferation), several growth factors can be included. These types of compositions may be "incorporated" as others, by inclusion in the composition or by co-administration in the present methods. Examples include vascular endothelial growth factor (VEGF), transforming growth factor (TGF), such as transforming growth factor p, platelet derived growth factor (PDGF), epidermal growth factor (EGF), nerve growth factor (NGF), insulin-like growth factor (e.g. insulin-like growth factor I), glial growth factor (GGF), fibroblast growth factor (FGF), etc. It should be noted that in many cases these terms refer to many different molecules. For example, several known molecules of transforming growth factors R are known in the art. One skilled in the art will be guided in the selection of an appropriate growth factor, taking into account, for example, the place where the composition is to be administered. For example, EGF may be included in compositions applied to the skin; NGF and / or GGF may be included in compositions applied to the nerves or nervous system; and so on. For example, EGF may be included in compositions applied to the skin; NGF and / or GGF may be included in compositions applied to the nerves or nervous system; and so on. For example, EGF may be included in compositions applied to the skin; NGF and / or GGF may be included in compositions applied to the nerves or nervous system; and so on.

[0087] The growth factor or other agent can be a chemotactic substance that is in vivo or cell culture able to recruit cells to the site where the substance is present. Recombinant cells can potentially contribute to the formation of new tissue or repair existing damaged tissue (e.g., by structurally and / or functionally contributing to the tissue (e.g., by delivery of growth factors or contributing to the desired immune response)). Some chemotactic substances may also act as proliferating agents (e.g., neurotropic agents such as NGF or BDNF).

[0088] The compositions may also be used in combination with or instead of compounds such as cyanoacrylates, oxidized cellulose, fibrin sealants, collagen gel, thrombin powder, microporous powdered polysaccharides, coagulation factors (e.g., factor V, factor VIII, fibrinogen or prothrombin) and powdered zeolites.

[0089] In one embodiment, vitamins can be added to self-assembling peptides such as vitamin K after liver surgery. In addition, other vitamins may be added to facilitate the reconstitution of tissue or skin when applied topically in combination with the material. This can occur after injury or in the normal local hydration mode.

[0090] One or more therapeutic, diagnostic and / or prophylactic agents may be administered concurrently with self-assembling peptides in the same formulation, may be administered simultaneously in separate formulations or sequentially. Alternatively, the active agent (s) may be covalently coupled to the self-assembling peptide.

[0091] It is understood that the therapeutic molecules are generally administered in an effective amount to achieve a clinically relevant result, and effective doses and concentrations are known in the art. These doses and concentrations may lead to selection of doses and concentrations in the present context. The bioactive molecules can be delivered in a variety of suitable concentrations and in appropriate amounts (e.g., in the microgram range or milligram range, or more). For guidance, you can read the texts such as Pharmacological Basis of Therapeutics Goodman and Gilman, ed. 10., and Katzung, Basic and Clinical Pharmacology.

Cells [0092] When cells are delivered to a patient (e.g., to promote tissue healing), autologous cells can be used. In one embodiment, the cells may be hematopoietic cells from a patient, dispersed in self-assembling peptides, and implanted. In another embodiment, the cells may be red blood cells from umbilical cord blood.

[0093] Molded scaffolds as described above, liquid compositions, gels, solids (e.g., powders) or other semisolid forms may contain one or more additional substances, such as bioactive molecules or cells. In some cases, a cell may secrete a bioactive molecule either naturally or by using genetic engineering methods (e.g., expression and / or secretion of a recombinant protein). The structures described herein are able to support adhesion, viability and cell growth; they were observed when the cells were grown on the surface of the material or when the cells grow in the material (e.g. when they are encapsulated). In addition, structures can serve as substrates for neurite growth and synapse formation when neurons grow on or inside them. Therefore,

09/778200 and 10/196942).

E. Formulations [0094] In a preferred embodiment, the formulation is a liquid or reconstitutable powder applied topically. In one embodiment, the formulation is provided as a dry or lyophilized powder that can be administered directly as a powder that hydrates at the application site or can be suspended or dissolved in a liquid, most preferably aqueous and used in the form of a spray or injection or hydrogel, such as chitin, collagen, alginate or synthetic polymer. In another embodiment, the formulation is administered in the form of a compressed wafer, disk or tablet. In yet another embodiment, the formulation is provided as a coating on a device, e.g. a stent or catheter, which can be dissolved in an aqueous solution and dried on the device, or mixed with a polymeric carrier and applied to the device. In yet another embodiment, the formulation is provided in the form of a bandage, foam or template in which the peptides can be dispersed or absorbed. The formulation may also be in the form of sutures, tape or glue.

[0095] Typically, local anesthetics are delivered by topical administration (e.g., formulated as an ointment, cream or solution) or injected into the area where the nerve fibers are to be blocked. The formulation may be administered for a burn or an ulcer, especially in the case of a preparation containing anesthetics, anti-inflammatory agents, growth factors and anti-infective agents, in the form of foam, matrix or bandage, to stop bleeding or loss of interstitial fluid.

[0096] One or more of the compositions described herein may be assembled in kits containing instructions for use. For example, the kits may include a biocompatible composition comprising self-assembling peptides (or a concentrated solution thereof or powdered formulation together with a diluent) and a vasoconstrictor, a colorant or painkiller, or anesthetic and instructions for combining them (provided they have not already been combined) and use (e.g., dilution and administration). The kits may further comprise one or more additional measures described herein. These agents may be present in the peptide-based composition or may be packaged separately and may include one or more of a type of biological cell, antibiotic or other therapeutic agent, collagen, an anti-inflammatory agent, a growth factor or a nutrient. The kit may also contain one or more syringes (e.g., a cylindrical syringe or a medical syringe), a needle, a pipette, gauze, sponge or cotton swab, swabs, bandage, nasal bleeding plug, disinfectant, surgical sutures, scissors, scalpel, sterile fluid, spray container, including those in which the liquid solution is sprayed by a simple hand pump, a sterile container or disposable gloves.

[0097] The formulation may be administered as suitable for the treatment of one or more disorders. For example, the formulation may be used to repair wounds or during surgery of the lung or dura mater, or after epidural or lumbar puncture, in order to stop the leakage of the cerebrospinal fluid. The preparation may be dispersed in a seam or adhesive for administration over time or be released after sewing or gluing the wound, thus limiting bleeding, loss of body fluids or other fluids such as those generated by parenchymal tissues, such as the liver, pancreas and gastrointestinal tract. The preparation can be used anywhere in the bleeding, in bandages, in a gas, sponge or other material, to immediately control the bleeding, or it can be released later to control the bleeding, if the initial treatment, such as sewing or pressure, is insufficient. Dried fabrics, dehydrated foams or hydrogels, or bandages containing the preparation may be part of first aid kits for the treatment of wounds, for example, during war, in places of accident or in clinics where rapid treatment may be required, and the storage space is limited .

[0098] In some embodiments, compositions comprising self-assembling peptides may be associated with surgical sponges. For example, liquid compositions can be sucked into commercially available sponges before or during their use. Studies indicate that hemostasis can be achieved satisfactorily without traditional sponges, but there may be instances in which compositions containing self-depositing material may be beneficial (e.g., when the patient has profound bleeding or when the goal of treatment is temporary stabilization). The compositions used may contain any of the non-fibrous agents described herein. The sponges may be any known in the art, including woven and non-woven sponges and those specifically intended for dental or ophthalmic surgery. See, e.g., US Pat. Nos. 4098728; 4211227; 4636208; 5180375; and 6711879.

[0099] In embodiments involving bandages or dressings, the bandage or dressing may include a first layer of sufficient shape and size to cover the wound, or a substantial portion thereof (e.g., the most damaged part of the tissue or the most bleeding area). The first layer may have a top surface, a bottom surface and a rim that is optionally, completely or partially covered with glue. The second layer of bandage or dressing may be releasably attached to the bottom surface of the first layer, optionally excluding the periphery or any part of the perimeter containing the adhesive, and may include a liquid or non-liquid composition (e.g., gel, paste, foam, cream, ointment or powdered composition) containing self-assembling peptides. The composition comes in contact with the wound after the application of the bandage or dressing and can be transferred from the bandage or dressing to the wound site after the removal of the first or first and second layer. In simpler arrangements, a composition comprising self-assembling peptides may be associated with the lower part of the first layer (e.g., inside the adhesive edge) and the second layer may be omitted. In both cases, the first and / or the second layer may comprise a transparent window through which one or more of the underlying wounds can be viewed. The composition containing self-assembling peptides can be added to the bandages before they are packaged or just before use. In another embodiment, the formulation may contain another physical barrier, such as a silicone film layer, to prevent loss of fluid as a result of drying, after

[0100] The formulations may also be administered as immediate or controlled release preparations. A delayed release dosage form is one that releases the drug (or drugs) at a different time than immediately after administration. The sustained release dosage form is one that allows at least a two-fold reduction in the dosage frequency when compared to a drug presented in the form of a conventional dosage form (e.g., as a solution or a conventional solid dosage form with rapid drug release). The modified release dosage form is one for which the time, route and / or drug release characteristics are selected to achieve a therapeutic or convenient target that is not provided by conventional dosage forms such as solutions, ointments or rapidly dissolving forms. dosed.

[0101] The matrix-forming materials are materials that form strong, viscous gels upon hydration and provide control of diffusion and drug release. In hydrophilic matrix systems, the matrix forming materials are uniformly incorporated into the whole tablet. In contact with water, the outer layer of the tablet is partially hydrated to form a gel layer. The rate of diffusion of the drug (s) from the gel layer and the rate of erosion of the gel layer determines the complete dissolution of the tablets and the rate of drug delivery. Examples of matrix forming materials include cellulose ethers which are water-soluble such as methylcellulose, ethylcellulose and hydroxypropylmethylcellulose.

[0102] The formulations are prepared using a pharmaceutically acceptable "barrier" consisting of materials that are believed to be safe and effective and can be administered to the subject without causing undesirable biological side effects or undesirable effects. "Vehicle" means all components present in a pharmaceutical preparation other than the active ingredient (s). The term "carrier" includes, but is not limited to, diluents, binders, lubricants, disintegrants, fillers, matrix-forming compositions, and coating compositions.

[0103] "Carrier" also includes all components of a coating composition that may include plasticizers, pigments, colorants, stabilizing agents, and lubricants. The delayed release dosage formulations can be prepared as described in the references, such as "Pharmaceutical dosage form tablets", edited by Liberman et al. (New York, Marcel Dekker, Inc., 1989), "Remington - The science and practice of pharmacy," ed. 20., Lippincott Williams &amp; Wilkins, Baltimore, Md., 2000, and "Pharmaceutical dosage forms and drug delivery systems," ed. 6., Ansel et al., (Media, Pa .: Williams and Wilkins, 1995), which provide information on carriers, materials, equipment and methods for making tablets and capsules and delayed release dosage forms in the form of tablets, capsules and granules. .

[0104] Examples of suitable coating materials include, but are not limited to, cellulosic polymers, such as cellulose acetate phthalate, hydroxypropylcellulose, hydroxypropylmethylcellulose, hydroxypropylmethylcellulose phthalate, and hydroxypropylmethylcellulose acetate succinate; poly (vinyl acetate phthalate), acrylic acid polymers and copolymers, and methacrylic resins, which are commercially available under the trade name Eudragit ™ (Roth Pharma, Westerstadt, Germany), Zein, shellac and polysaccharides. In addition, the coating material may contain conventional carriers such as plasticizers, pigments, colorants, lubricants, stabilizing agents, pore forming agents and surfactants. Optional pharmaceutically acceptable excipients present in tablets, beads, granules or particles containing the drug include, but are not limited to,

[0105] Diluents, also referred to as "fillers", are usually necessary to increase the mass of the solid dosage form so as to provide a practical size for compressing the tablets or forming beads and granules. Suitable diluents include, but are not limited to, dicalcium phosphate dihydrate, calcium sulfate, lactose, sucrose, mannitol, sorbitol, cellulose, microcrystalline cellulose, kaolin, sodium chloride, dry starch, hydrolysed starch, pregelatinized starch, silicon dioxide, titanium oxide, aluminosilicate magnesium and powdered sugar.

[0106] Binders are used to confer cohesive properties on a solid dosage formulation, and thus ensure that the tablet or spheres or granule will remain intact after the dosage forms have been formed. Suitable binding materials include, but are not limited to, starch, pregelatinized starch, gelatine, sugars (including sucrose, glucose, dextrose, lactose and sorbitol), polyethylene glycol, waxes, natural and synthetic gums such as gum arabic, tragacanth, alginate sodium, cellulose, including hydroxypropylmethylcellulose, hydroxypropylcellulose, ethylcellulose and veegum, and synthetic polymers, such as copolymers of acrylic acid and methacrylic acid, methacrylic acid copolymers, methyl methacrylate copolymers, aminoalkyl methacrylate copolymers, poly (acrylic acid) / poly (methacrylic acid) and polyvinylpyrrolidone.

[0107] Lubricants are used to facilitate the preparation of tablets. Examples of suitable lubricants include, but are not limited to, magnesium stearate, calcium stearate, stearic acid, glycerol behenate, polyethylene glycol, talc, and mineral oil. [0108] Disintegrants are used to facilitate the disintegration of the dosage form or "disintegration" after administration and include, but are not limited to, starch, sodium starch glycolate, sodium carboxymethyl starch, carboxymethylcellulose sodium, hydroxypropylcellulose, pregelatinized starch, clays, cellulose, alginine, gums or cross-linked polymers, such as cross-linked PVP (Polyplasdone ™ XL from GAP Chemical Corp).

[0109] Stabilizers are used to inhibit or delay drug decomposition reactions that include, for example, oxidation reactions.

[0110] Surfactants may be anioncationic, amphoteric or nonionic surfactants. Suitable anionic surfactants include, but are not limited to, those containing carboxylate, sulfonate and sulfate ions. Examples of anionic surfactants include the sodium, potassium, ammonium, and ammonium salts of long-chain alkylsulfonates and alkylaryl sulfonates, such as sodium dodecylbenzenesulfonate; sodium dialkyl sulfosuccinates, such as sodium dodecylbenzenesulfonate; sodium dialkyl sulfosuccinates, such as sodium bis (2-ethylthioxyl) sulfosuccinate; and alkyl sulfates, such as sodium lauryl sulphate. Cationic surfactants include, but are not limited to, quaternary ammonium compounds such as benzalkonium chloride, benzethonium chloride, cetrimmonium bromide, stearyl-dimethyl benzyl ammonium chloride, polyoxyethylene and coconut amine. Examples of non-ionic surfactants include ethylene glycol monostearate, propylene glycol myristate, glycerol monostearate, glycerol stearate, polyglycerol-4 oleate, sorbitan acylate, sucrose acylate, PEG-150 laurate, PEG400 monolaurate, polyoxyethylene monolaurate, polysorbates, octylphenyl polyoxyethylene, cetyl ether PEG-1000, tridecyl polyoxyethylene ether, polypropylene glycol butyl ether, Poloxamer^™ 401, stearoyl monoisopropanolamide and a polyoxyethylene amide of hardened tallow. Examples of amphoteric surfactants include sodium N, N, N, N, N, N-dipropionate, myristoamphoacetate, laurylbetaine and lauryl sulfobetaine.

If desired, tablets, beads, granules or particles may also contain a small amount of non-toxic excipients, such as wetting or emulsifying agents, colorants, pH buffering agents, and preservatives.

[0112] In one type of preparation, the self-assembling peptides may be used as a shaving cream or liquid hand lotion, creating a barrier to fluid loss and as a barrier to adhesions and contamination.

[0113] Sustained release formulations are generally prepared as diffusion or osmotic systems, for example as described in "Remington-The science and practice of pharmacy" (20th Edition, Lippincott Williams &amp; Wilkins, Baltimore, MD, 2000) . The diffusion system typically consists of two types of devices, a reservoir and a matrix, and is well known and described in the art. Matrix devices are typically made by compressing the drug with a slowly dissolving polymeric carrier into a tablet. The three main types of materials used in the manufacture of matrix devices are insoluble plastics, hydrophilic polymers and fatty compounds. Plastic matrices include methyl acrylate-methyl methacrylate, polyvinyl chloride and polyethylene. The hydrophilic polymers include cellulosic polymers, such as methyl and ethylcellulose, hydroxyalkylcelluloses, such as hydroxypropylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose and Carbopol ™ 934, polyethylene oxides and mixtures thereof. Fatty compounds include, but are not limited to, different waxes, such as carnauba wax and glycerol tristearate, and wax-like substances, including hydrogenated castor oil or hydrogenated vegetable oil or mixtures thereof. In certain embodiments, the plastic material is a pharmaceutically acceptable acrylic polymer, including but not limited to copolymers of acrylic acid and methacrylic acid, methyl methacrylate, methyl methacrylate copolymers, ethoxyethyl methacrylate, methacetyl methacrylate, aminoalkyl methacrylate copolymer, poly (acrylic acid), poly (methacrylic acid), methacrylic acid-alkylamine copolymer, poly (methyl methacrylate), poly (methacrylic acid) (anhydride), polymethacrylate, polyacrylamide, poly (methacrylic acid anhydride) and glycidyl methacrylate copolymers. In some embodiments, the acrylic polymer consists of one or more ammonium methacrylate copolymers. Ammonium methacrylate copolymers are well known in the art and described in NF XVII as fully polymerized copolymers of acrylic and methacrylic acid esters with a low content of quaternary ammonium groups.

[0114] Alternatively, sustained release formulations may be prepared using osmotic systems or by applying a semipermeable coating to the dosage form. In the latter case, the desired drug release profile can be achieved by combining in a suitable ratio a material with low permeability and high permeability.

[0115] The immediate release portion may be added to the sustained release system by applying an immediate release layer to the upper portion of the sustained release core using a coating or compression process, or in a multiple unit system such as a capsule-containing capsule. prolonged and immediate release. Tablets with sustained release containing hydrophilic polymers are prepared by methods commonly known in the art, such as direct compression, wet granulation or dry granulation. Their formulations usually contain polymers, diluents, binders and lubricants, as well as a pharmaceutical active ingredient. Usual diluents include inert powdered substances such as starches, powdered cellulose, especially crystalline and microcrystalline cellulose, sugars such as fructose, mannitol and sucrose, grain flours and similar edible powders. Typical diluents are, for example, various types of starch, lactose, mannitol, kaolin, phosphate or calcium sulphate, inorganic salts such as sodium chloride and powdered sugar. Powder cellulose derivatives are also useful. Typical tablet binding agents include substances such as starch, gelatin and sugars, such as lactose, fructose and glucose. Natural and synthetic gums, including gum arabic, alginates, methylcellulose and polyvinylpyrrolidone, can also be used. Polyethylene glycol, hydrophilic polymers, ethylcellulose and waxes may also serve as binders. A lubricant is needed in the tablet formulation so that the tablet and the punches do not adhere to the matrix. The lubricant is selected from such slippery solids as talc, magnesium and calcium stearate, stearic acid and hydrogenated vegetable oils. The prolonged-release tablets containing waxy materials are generally prepared by methods known in the art, such as a direct mixing method, a solidification method, and a method using an aqueous dispersion. In the solidification process, the drug is mixed with the wax material and solidifies or solidifies and sieves and processes.

[0116] Preferred coating weights for individual coating materials can be readily determined by those skilled in the art, assessing individual release profiles for tablets, beads and granules made with varying amounts of different coating materials. It is a combination of materials, methods and forms of application that give the desired release characteristics that can be determined only on the basis of clinical studies. The coating composition may contain conventional additives such as plasticizers, pigments, colorants, stabilizers, lubricants, etc. A plasticizer is typically present to reduce the friability of the coating and is generally about 10 wt.%. up to 50% by weight relative to the dry weight of the polymer. Examples of typical plasticizers include polyethylene glycol, propylene glycol, triacetin, dimethyl phthalate, diethyl phthalate, dibutyl phthalate, dibutyl sebacate, triethyl citrate, tributyl citrate, triethylacetyl citrate, castor oil and acetylated monoglycerides. Preferably, a stabilizing agent is used to stabilize the particles in the dispersion. Typical stabilizing agents are nonionic emulsifiers, such as sorbitan esters, polysorbates and polyvinylpyrrolid. It is recommended to use lubricants to reduce the effect of sticking during film formation and drying, and generally make up about 25 wt.%. up to 100% by weight polymer mass in the coating solution. One effective lubricant is talc. Other glidants, such as magnesium stearate and glycerol monostearates, may also be used. Pigments such as titanium dioxide may also be used.

Polymer matrices [0117] Both non-biodegradable and biodegradable matrices may be used to deliver self-assembling peptides, although biodegradable matrices are preferred. These may be natural or synthetic polymers, although synthetic polymers are preferred due to better characteristics of degradation and release profiles. The polymer is selected based on the period in which release is required. In some cases, linear release may be most useful, although in others the pulsed release or "volumetric release" may provide more effective results. The polymer may be in the form of a hydrogel (typically up to about 90% by weight of water) and optionally crosslinked with multivalent ions or polymers.

[0118] Representative synthetic polymers that can be used for delivery include polyamides, polycarbonates, polyalkylenes, polyalkylene glycols, polyalkylene oxides, polyalkylene terephthalates, polyvinyl alcohols, polyvinyl ethers, poly (vinyl esters) vinyl), polyvinyl halides, polyvinylpyrrolidone, polyglycolides, polysiloxanes, polyurethanes and copolymers, alkylcellulose, hydroxyalkylcelluloses, cellulose ethers, cellulose esters, nitrocelluloses, acrylic and methacrylic esters, methylcellulose, ethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, hydroxybutylmethylcellulose, cellulose acetate , cellulose propionate, cellulose acetate butyrate, cellulose acetate phthalate, carboxyl ethylcellulose, cellulose triacetate, cellulose sulfate sodium salt, poly (methyl methacrylate), poly (ethyl methacrylate),poly (butyl methacrylate), poly (isobutyl methacrylate), poly (hexylmethacrylate), isodecyl polyacrylate, poly (lauryl) methacrylate, poly (phenylmethacrylate), poly (methyl acrylate), poly (isopropyl) acrylate, poly (isobutyl acrylate), poly (octadecyl acrylate), polyethylene, polypropylene, polyethylene glycol, poly (ethylene oxide), poly (ethylene terephthalate), poly (vinyl alcohols), polyvinyl acetate, poly (vinyl chloride), polystyrene and polyvinylpyrrolidone.polystyrene and polyvinylpyrrolidone.polystyrene and polyvinylpyrrolidone.

[0119] Examples of non-biodegradable polymers include ethylene-vinyl acetate, poly (meth) acrylic acid), polyamides, copolymers and mixtures thereof. Examples of biodegradable polymers include synthetic polymers, such as polymers of lactic acid and glycolic acid, polyanhydrides, poly (ortho) esters, polyurethanes, poly (butylic acid), poly (valeric acid) and poly (lactide-co-caprolactone) and natural polymers, such as alginate and other polysaccharides, including dextran and cellulose, collagen, their chemical derivatives (by substitution, additions of chemical groups, e.g., alkyl, alkylene, hydroxylation, oxidation and other modifications routinely performed by those skilled in the art), albumin and others hydrophilic proteins, zein and other prolamins and hydrophobic proteins, their copolymers and mixtures.

[0120] Bioadhesive polymers of particular interest include bioerodible hydrogels described by HS Sawhney, CP Pathak and JA Hubell in Macromolecules, 1993, 26, 581-587, poly (hyaluronic acids), casein, gelatin, glutin, polyanhydrides. , poly (acrylic acid), alginate, chitosan, poly (methyl methacrylate), poly (ethyl methacrylate), poly (butyl methacrylate), isobutyl poly (meth) acrylate, poly (hexylmethacrylate), poly (isodecyl) methacrylate, poly (methacrylate) lauryl), poly (phenylmethacrylate), poly (methyl acrylate), poly (isopropyl) acrylate, poly (isobutyl) acrylate and poly (octadecyl acrylate).

[0121] The matrix may be in the form of microparticles, such as microspheres, where the peptides are dispersed in a solid polymer matrix or microcapsules, wherein the core is of a different material from the polymer shell and the peptide is dispersed or suspended in a core that may be liquid or permanent. Unless specified herein in detail, the microparticles, microspheres and microcapsules are used interchangeably herein. Alternatively, the polymer may be cast in the form of a thin plate or foil ranging from nanometers to four centimeters, powder made by milling or other standard techniques, and even a gel, such as a hydrogel. The polymer may also be in the form of a coating or part of a stent or catheter, vascular graft or other prosthetic device. [0122] The matrices may be formed by evaporation of the solvent,

[0123] Bioerodible microspheres can be prepared using any methods developed for the production of drug delivery microspheres, for example as described in Mathiowitz and Langer, J. Controlled Release 5,13-22 (1987); Mathiowitz et al., Reactive Polymers 6, 275-283 (1987); and Mathiowitz, et al., J. Appl. Polymer Sci. 35, 755-774 (1988). The choice of the method depends on the choice of polymer, its size, external morphology and desired crystallinity, as described for example by Mathiowitz, et al., Scanning Microscopy 4,329-340 (1990); Mathiowitz, et al., J. Appl. Polymer Sci. 45, 125-134 (1992); and Benita, et al., J. Pharm. Sci. 73, 1721-1724 (1984). In solvent evaporation, as described for example in Mathiowitz et al. (1990), Benita and US Patent No. 4,273,293, Jaffe, the polymer is dissolved in a volatile organic solvent. The peptide in soluble form or dispersed in the form of fine particles is added to the polymer solution and the mixture is suspended in an aqueous phase that contains a surfactant, such as polyvinyl alcohol. The emulsion obtained is mixed until most of the organic solvent evaporates, leaving the solid microspheres. In general, the polymer can be dissolved in methylene chloride. Microspheres of various sizes (1-1000 microns) and different morphologies can be obtained by this method, which is useful for relatively stable polymers such as polyesters and polystyrene. However, unstable polymers, such as polyanhydrides, can degrade as a result of exposure to water. For these polymers, preferably hot melt encapsulation and solvent removal are used.

[0124] In hot melt encapsulation, the polymer is first melted and then mixed with solid peptide particles. The mixture is suspended in an immiscible solvent, such as silicone oil and, while stirring, it is heated to room temperature

5 ° C above the melting point of the polymer. After the emulsion has stabilized, it is cooled until the polymer particles solidify. The resulting microspheres are washed by decanting with petroleum ether to obtain a free flowing powder. In this method, microspheres with diameters between one and 1000 microns can be obtained. The outer surface of the spheres created with this technique is usually smooth and dense. This procedure is useful for water-labile polymers, but is limited to use with polymers with a molecular weight between 1,000 and 50,000. Solvent removal has primarily been designed for use with polyanhydrides. In this method, the drug is dispersed or dissolved in a solution of the selected polymer in a volatile organic solvent, such as methylene chloride. The mixture is then suspended in an oil such as silicone oil while stirring to form an emulsion. Within 24 hours, the solvent diffuses in the oil phase, and the emulsion droplets cure to form solid polymer microspheres. In contrast to solvent evaporation, this method can be used to produce microspheres from polymers with a high melting point and a wide molecular weight range. In this procedure, microspheres with diameters between one and 300 microns can be obtained. The external morphology of the spheres depends to a large extent on the type of polymer used. On spray drying, the polymer is dissolved in methylene chloride (0.04 g / ml). A known amount of the active drug is suspended (if it is insoluble) or simultaneously dissolved (if it is soluble) in the polymer solution. The solution or dispersion is then spray-dried. Dihedral microspheres can be prepared in accordance with US Pat. No. 4,866,277 to Mathiowitz.

[0125] Hydrogel microspheres made from gel-type polymers, such as alginate or polyphosphites or other dicarboxylic polymers, can be prepared by dissolving the polymer in an aqueous solution, suspending the material to be introduced into the mixture, and extruding the polymer mixture through a molding device. microdroplets, which is equipped with a stream of nitrogen gas. The resulting microspheres fall into a slowly stirred, ionic curing bath as described, for example, by Salib et al., Pharmazeutische Industrie 40-11A, 1230 (1978). Chitosan microspheres can be prepared by dissolving the polymer in an acidic solution and crosslinking with a tripolyphosphate. For example, carboxymethylcellulose (CMC) microspheres are prepared by dissolving the polymer in an acid solution and precipitating the microspheres with lead ions.

[0126] Other delivery systems including films, coatings, granules, cachets and devices can be made using dipping, melt casting and extrusion molding as well as standard composites manufacturing methods. The polymer can be produced by first mixing monomers and peptides as described in Sawhney et al. And polymerizing the monomers using UV light. The polymerization can be carried out in vitro as well as in vivo.

F. Delivery devices [0127] Liquid preparations may be provided in a syringe or pipette having a cylinder containing a composition comprising self-depositing peptides and means for ejecting the composition from the open end of the syringe or pipette (e.g., plunger or bulb). The syringe may consist of one or more compartments such that the mixing of self-assembling peptides with one or more other agents takes place at the time of use. The compartments may also contain an excipient, such as a hydrogel forming material or glue in one compartment, and self-assembling peptides in another compartment. In another embodiment, one compartment may comprise a lyophilized powder or particles of self-assembling peptides, and another compartment may include a solution for dissolving or hydration of peptides or other powders for mixing with self-assembling peptides for use in a dry form. The composition in the cylinder may further comprise any non-fibrous agent described herein (e.g., one or more of a vasoconstrictor, a coloring agent, an anesthetic or analgesic, an antibiotic or other therapeutic agent, collagen, anti-inflammatory agent, growth factor or nutrient).

[0128] Self-assembling peptides can be applied as a coating by spraying or immersing the device in peptides, peptides can be impregnated with bandages, gases or other absorbent material, the peptides can be mixed with the polymeric material. Self-assembling peptides can also be formulated as a pharmaceutical foam. Pharmaceutical foams are pressurized dosage forms that, during actuation of the valve, emit fine dispersions of liquid and / or solid materials in a gaseous medium. In one embodiment, the foam comprises self-assembling peptides, in liquid or solid form, optionally in combination with one or more active agents. Suitable propellants include, but are not limited to, hydrofluoroalkanes (HFAs) such as 1,1,1,2-tetrafluoroethane (HFA 134a) and 1,1,1,2,3,3,3-heptafluoropropane (HFA 227), hydrocarbons and carbon dioxide.

III. Methods of administration

A. Administration sites [0129] Self-assembling peptides can be applied to various surfaces to prevent or control the passage of the fluid or act as a barrier. The amount of self-assembling peptides is determined in part by the function of peptides in controlling fluid flow, as well as the properties of any other materials or structures associated with supporting peptides, alone or in combination with other bioactive substances. Self-assembling peptides can be used to stop the movement of fluids into or out of tissues / organs.

[0130] In a first embodiment, self-assembling peptides are used to prevent or control bleeding. Self-assembling peptides can be applied in the form of a powder, liquid, gel or as part of a substrate, such as a bandage or membranes. They can be applied to the blood vessel, to the light, for example, during angioplasty, administered by or as a coating on the stent or catheter, or on the outer side of the vessel, usually at the site of anastomosis. Self-assembling peptides can be applied to tissues before, during or after surgery to prevent bleeding, which is a particular problem for tissues, such as the liver, kidney or spleen, or other surgical procedures where there is a high risk of transfusion or to seal and protect the tissue, for example, which is intended for transplantation or re-connection. In another embodiment, the self-assembling peptides can be used as an additive to a shaving cream in which they can act as a hemostatic means to stop bleeding due to jams on the razor, as a barrier to prevent contamination at the razor jams and / or as a lubricant.

[0131] Self-assembling peptides may be used to stop the flow of fluids other than blood. Self-assembling peptides can be applied to burns to prevent leakage of interstitial fluid. Self-assembling peptides can be applied to the dura mater or lung as a tire or pulmonary sealant. In one embodiment, the self-assembling peptides can be used to repair the lung on the stab wound, thereby restoring its ability to function.

[0132] Self-assembling peptides can also be used as a barrier to oral surgery, periodontics and general dentistry.

[0133] Use of self-accumulating peptides in subjects with coagulation disorder (haemophilia, von Willebrand disease, vitamin K deficiency, protein S or protein C, fulminant hepatitis, disseminated intravascular coagulation syndrome ("DIC"), haemolytic-uremic syndrome ("HUS" ")) Is also of considerable utility, since the mechanism of action is independent of the correct solidification path. For example, the compositions can be used to replace a damaged semipermeable barrier, such as in cells, to restore the local environment and to facilitate cell survival and repair.

[0134] In another embodiment, self-assembling peptides are typically applied by spraying or injecting onto the external surface of a tissue, such as a tumor, to prevent disintegration or metastasis during surgery. Self-assembling peptides control bleeding during tumor resection and also reduce metastasis. It also minimizes the immune response that a laser can cause during tumor resection. Self-assembling peptides are also useful for keeping loose tumors together, so that nothing is left when they are removed. There are several types of cancer that are difficult to resect, because they do not stick tightly together (ie they are not solid masses). It is anticipated that self-assembling peptides are particularly useful in tumor resection in the brain and may be useful in dose response for subcutaneous tumor resection. This may facilitate the resection of melanomas in the skin, as it turns out that self-assembling peptides also facilitate skin healing. In addition, self-assembling peptides comprising a tumor-specific targeting segment can be attached to the tumor, resulting in tumor aggregation, allowing resection of the tumor. Self-assembling peptides can also immobilize cells that detach from the tumor during resection to retain or slow down the metastases. Self-assembling peptides may also include a reactive marker with certain types of antigens on the surface of tumor cells, causing a colorimetric change to show that all cells have been removed or that there are still cells that should be removed. Adding the indicator to self-assembling peptides, as well as the ability of the peptides to act as a biological marker, may reduce the necessity of the second and third operations as well as the complications associated with external contamination in the operating field. Self-assembling peptides can also be used to deliver materials, such as DNA, to a wound site for a long period of in vitro and for many in vivo treatments. A further advantage of self-assembling peptides is that they can be injected and gelled in place so that the peptides can be applied and reapplied during surgery, if necessary. as well as the ability of the peptides to act as a biological biomarker may reduce the necessity of the second and third operations as well as the complications associated with external contamination in the operating field. Self-assembling peptides can also be used to deliver materials, such as DNA, to a wound site for a long period of in vitro and for many in vivo treatments. A further advantage of self-assembling peptides is that they can be injected and gelled in place so that the peptides can be applied and reapplied during surgery, if necessary. as well as the ability of the peptides to act as a biological biomarker may reduce the necessity of the second and third operations as well as the complications associated with external contamination in the operating field. Self-assembling peptides can also be used to deliver materials, such as DNA, to a wound site for a long period of in vitro and for many in vivo treatments. A further advantage of self-assembling peptides is that they can be injected and gelled in place so that the peptides can be applied and reapplied during surgery, if necessary. such as DNA, to a wound site for a long period of in vitro and for many in vivo treatments. A further advantage of self-assembling peptides is that they can be injected and gelled in place so that the peptides can be applied and reapplied during surgery, if necessary. such as DNA, to a wound site for a long period of in vitro and for many in vivo treatments. A further advantage of self-assembling peptides is that they can be injected and gelled in place so that the peptides can be applied and reapplied during surgery, if necessary.

[0135] In yet another embodiment, the self-assembling peptides are particularly well adapted to act as a barrier to prevent contamination into the tissue or tissue, for example during intestinal surgical treatment. Self-assembling peptides can be used to prepare an internal site before surgery, especially in places such as sinus cavities and for surgical procedures such as transurethral and transvaginal surgery. Self-assembling peptides should also be particularly useful in cardiovascular surgery, where both barrier and hemostatic properties may be valuable, for example in heart valve patients who are susceptible to adverse consequences, such as ring valve abscess (cover with a valve cover). add antibiotic), endocarditis (cover the valve), dissection of the aortic root (provide immediate hemostasis). In yet another embodiment, a mixture of complementary amino acid sequences that are not self-assembling can be applied to the tissue to block infection of the tissue by the bacteria. The fact that the sequences are complementary should result in a more effective barrier than complementary sequences that self-assemble.

[0136] Self-assembling peptides in combination with a metal, such as silver, have anti-adhesive properties and can inhibit angiogenesis. Accordingly, they may be useful in reducing scarring and adhesions. Self-assembling peptides are applied after surgery or injury, such as burns, to reduce scarring, fluid loss and reduce infection. This is further used in plastic surgery, in particular to protect clean and debris-free areas before skin closure or transplantation, for example, in abdominoplasty, facial skin lift, transplantation sites, in the case of the latissimus dorsal muscle for breast reconstruction.

[0137] In yet another embodiment, self-assembling peptides are administered as a suspension that the patient can drink to reduce gastric bleeding, for example from an ulcer or to reduce acidity. Alternatively, the self-assembling peptides may be provided in the form of an enema or suppository for the treatment of haemorrhoids or for the filling of diverticulas. In yet another embodiment, self-assembling peptides may be used to prevent infertility due to adhesions in the fallopian tubes or vectors.

[0138] Self-assembly is not irreversible, the contained substances can be released. For example, molecules or cells can be released from in vivo structures (e.g., small molecules can diffuse, and larger molecules and cells can be released when structures are degraded).

[0139] In yet another embodiment, self-assembling peptides are used as neuroprotective agents to minimize damage and scarring after nerve damage. The peptide-based structures promote the repair and regeneration of nerve tissue (e.g., when self-assembling peptides are applied to a brain lesion as described in USSN 10/968790). The small size of fibers in the scaffolding and / or open structure of the "weave" of materials allows for prolonging cellular processes and enables the proper diffusion of nutrients and wastes in a way that provides unique benefits for regeneration of nerve tissue.

[0140] In the process of repairing wounds, the compositions may not only improve the final result (e.g., reduced scar formation, leading to a result that more closely resembles the original tissue), but also reduce the time required for healing. These results could not be predicted from the results obtained after being used for wounds within the central nervous system, taking into account the significant differences between nervous and non-neuronal tissues.

[0141] Finally, self-assembling peptides can be used as a "nanosecure" to prevent cross-contamination. For example, self-assembling peptides can be applied as a coating outside the body and then self-assembled. Peptides that have self-deposited can stop the movement of liquid towards the interior of the body, thereby reducing the risk of cross-contamination.

B. Effective Doses [0142] In general, the amount of self-assembling peptides required depends on many factors, such as the size or extent of wounding (which in turn can be expressed in terms of length of incision, size or number of damaged blood vessels, degree of burns, size and depth of ulcer, abrasions or other wounds). The amount can vary, for example, from a few microliters to a few milliliters or more, e.g. tens or hundreds of milliliters. The device used to deliver self-assembling peptides will vary depending on the amount. For example, the syringe may conveniently be used to provide smaller amounts, while the tube or squeeze bottle may be more suitable for larger quantities. Effective amount (whether in relation to the scaffolding, its precursors, or another bioactive molecule present in the formulation), means an amount necessary to induce an improved or desired biological response. [0143] It will be appreciated by those of ordinary skill in the art that an effective amount of self-assembling peptides may vary depending on such factors as the desired biological endpoint, the peptides provided, the nature of the site to which the peptides are provided, and the type of condition due to whose agent is administered. For example, an effective amount of a composition for accelerating haemostasis may be an amount sufficient to reduce the amount of blood lost between the time of onset of bleeding and the time when the bleeding ends at at least 25% relative to the amount of blood lost after treatment with cold physiological saline or without treatment. An effective amount of a composition for accelerating haemostasis may also be an amount sufficient to reduce the time required to achieve arrest of visible bleeding of at least 25% relative to the time required after treatment with cold physiological saline or without treatment. An effective amount of a wound healing composition may be an amount sufficient to reduce the time required to achieve a predefined percentage reduction in lesion size of at least 25% relative to the time required in the absence of such treatment.

[0144] The amount of composition provided may vary depending on the severity of the subject's condition and should be sufficient to inhibit undesired movement to the extent that would benefit the subject. The body substance may be blood, cerebrospinal fluid, pus, serous exudate, bile, pancreatic juice or substance usually contained in the gastrointestinal tract (eg stomach or intestines) or urinary tract.

C. Method of Administration [0145] The composition may be delivered to the surface of the subject's body and / or delivered within a force generated cavity (e.g., by an unexpected injury or surgical procedure). In this way, undesirable movement of body substances can be inhibited in the context of a wide range of situations, including traumatic injury, health status (eg chronic or prolonged medical condition associated with bleeding) or surgical procedures (e.g., orthopedic surgery, dental surgery, cardiac surgery, ophthalmic surgery, plastic surgery or reconstructive surgery). For example, if the unwanted movement of the body substance is the result of an injury, the subject may have a partially or fully cut off part of the body, healing, abrasion or a stab wound. If the compositions are applied to the surface of the body, they can not only inhibit unwanted movement of the body substance, but also help to protect the individual against contamination. For example, the application of self-assembled peptides to the skin will impede the movement of undesirable foreign matter on the skin or bristles to the wound. If the unwanted movement of the bodily substance is associated with chronic disease, the subject may experience a relapse of bleeding. For example, the subject may be suffering from bleeding associated with varicose veins, including telangiectasias, hemorrhoids, bleeding in the lungs (e.g., from lung cancer, bronchitis or a bacterial or viral disease, including pneumonia or influenza) or esophageal varices. Medical conditions associated with recurrent bleeding can be treated using the compositions described herein, including those that comprise self-assembling peptides and a vasoconstrictor (e.g., phenylephrine, which may comprise about 0.25-0.5% of the composition). If the bleeding occurs in the throat or lungs, the compositions may be administered using a metered dose inhaler. If the patient's condition deteriorates to the point where artificial ventilation is required, the compositions may be administered via a ventilator or rinsing. [0146] Undesirable movement of the body substance may also occur during the surgical procedure, and the procedure may include an incision in the subject within the nervous system, eyes, ear, nose, mouth, throat, respiratory system, cardiovascular system, digestive system, urinary system, reproductive system, musculoskeletal system, liver or eyelid. The methods may be carried out regardless of whether the movement of the body substance was expedient. The compositions described herein can be used before or after undesired displacement (e.g., during a surgical procedure before the intended cut of the blood vessel or after unintentional cutting of the blood vessel). For example, the surgical procedure can be performed with the intention of repairing the aneurysm, inhibiting bleeding in the brain, treating esophageal varices, treating ulceration, or inhibiting loss of stomach contents or intestinal contents (e.g., from a swollen or ruptured appendage). The surgical procedure may involve the removal of a part of the intestine of the subject. Other procedures that can be performed with compositions containing self-assembling peptides include arteriography, cardiac catheterization, introduction of a stent, assistance in delivery by natural means or delivery by caesarean section, hysterectomy, organ transplantation, joint replacement or intervertebral disk excision. These procedures are representative. The surgical procedure can be performed by means of an endoscope or laparoscope, and the compositions can be delivered independently or from a chamber located inside these devices and connected to the distal end via a discharge channel to the subject's tissues. If the patient has an ulcer, the ulcer may be an esophageal, gastric, duodenal ulcer, diabetic or decubitus ulcer. More generally, the compositions can be applied to any damaged area of ​​the skin, and any of the methods described herein can include the step of identifying a subject in need of treatment. help in natural delivery or delivery by caesarean section, hysterectomy, organ transplantation, joint replacement or intervertebral disc excision. These procedures are representative. The surgical procedure can be performed by means of an endoscope or laparoscope, and the compositions can be delivered independently or from a chamber located inside these devices and connected to the distal end via a discharge channel to the subject's tissues. If the patient has an ulcer, the ulcer may be an esophageal, gastric, duodenal ulcer, diabetic or decubitus ulcer. More generally, the compositions can be applied to any damaged area of ​​the skin, and any of the methods described herein can include the step of identifying a subject in need of treatment. help in natural delivery or delivery by caesarean section, hysterectomy, organ transplantation, joint replacement or intervertebral disc excision. These procedures are representative. The surgical procedure can be performed by means of an endoscope or laparoscope, and the compositions can be delivered independently or from a chamber located inside these devices and connected to the distal end via a discharge channel to the subject's tissues. If the patient has an ulcer, the ulcer may be an esophageal, gastric, duodenal ulcer, diabetic or decubitus ulcer. More generally, the compositions can be applied to any damaged area of ​​the skin, and any of the methods described herein can include the step of identifying a subject in need of treatment. These procedures are representative. The surgical procedure can be performed by means of an endoscope or laparoscope, and the compositions can be delivered independently or from a chamber located inside these devices and connected to the distal end via a discharge channel to the subject's tissues. If the patient has an ulcer, the ulcer may be an esophageal, gastric, duodenal ulcer, diabetic or decubitus ulcer. More generally, the compositions can be applied to any damaged area of ​​the skin, and any of the methods described herein can include the step of identifying a subject in need of treatment. These procedures are representative. The surgical procedure can be performed by means of an endoscope or laparoscope, and the compositions can be delivered independently or from a chamber located inside these devices and connected to the distal end via a discharge channel to the subject's tissues. If the patient has an ulcer, the ulcer may be an esophageal, gastric, duodenal ulcer, diabetic or decubitus ulcer. More generally, the compositions can be applied to any damaged area of ​​the skin, and any of the methods described herein can include the step of identifying a subject in need of treatment. when the patient has an ulcer, the ulcer may be an esophageal, stomach, duodenal ulcer, diabetic or decubitus ulcer. More generally, the compositions can be applied to any damaged area of ​​the skin, and any of the methods described herein can include the step of identifying a subject in need of treatment. when the patient has an ulcer, the ulcer may be an esophageal, stomach, duodenal ulcer, diabetic or decubitus ulcer. More generally, the compositions can be applied to any damaged area of ​​the skin, and any of the methods described herein can include the step of identifying a subject in need of treatment.

[0147] A scaffold consisting of a self-assembling peptide (SAPNS) nanofiber may provide a transparent environment for the operative field, and also form an optically clear liquid that allows surgery through the resulting fluid / gel mixture. The surgical field is often covered with blood and residues during surgery. In addition, the removal of residues from the operating field usually requires washing the site with saline. Saline is only a temporary solution and should be used continuously to maintain a clean operating field. This creates several problems: any existing contamination is easily spread; a small hole will require alternating irrigation and operation; and during intestinal surgery, the use of saline may lead to serious infections causing postoperative complications. Applying SAPNS to biological limitations will reduce postoperative complications in endoscopic and open surgical procedures. Efficacy has been demonstrated on the brain, spinal cord, gastrointestinal tract, liver, muscles, arteries and veins.

[0148] For example, partial resection is currently performed as follows. Chi15 rurgine performs partial intestinal resection to remove the pre-cancerous area. An incision is made and the intestines are gently pulled out of the peritoneal cavity and placed on the table next to the patient. The defective area is cut out, and the two ends of the intestine are then joined together. Before putting the intestines back inside the body, the colostomy bag connects to the upper end of the intestine, and the operation area disinfects. The intestines are placed again in the abdominal cavity and then stitched. The drain is placed in the abdominal cavity to ensure there are no leaks or bleeding. In contrast, when using self-assembling peptides, a partial resection is performed as follows. The doctor opens the abdominal cavity and finds the violated part of the intestine. It is isolated using an additional liquid that is poured into the peritoneal cavity to isolate it from the rest of the peritoneal cavity. The surgeon reaches out through a gel that was created by the liquid and cuts the intestine. The two ends are connected to each other and the area is checked for any changes in color.

The gel also contains an indicator dye that changes to blue if there is a leak of gastric fluids or bacteria. The entire blue color is removed by suction. A few more self-assembled peptides are sprayed around the right area and the abdominal cavity is sutured.

[0149] Scar treatment: Experiments have shown that the application of a self-depositing material can be used to block the formation of scarring in the central nervous system (CNS). Administration of self-assembling peptides at the lesion site blocks stable scar formation, which allows regeneration at this site;

removing the scar that occurs in the central nervous system (CNS) allows axons to grow across the wound.

[0150] Wound healing enhanced by chelation: Self-assembling peptides can be used to deliver a chelating agent, such as iron, to a site, so that they can be used by the body in the local environment to regenerate the basal membrane. In tissues that do not contain enough iron, the supply of iron in a permanent form will help heal and rebuild the tissue. Metals with a cystine or cystine-like residue may be incorporated into the nanomaterial such that there is little or no steric hindrance to the matrix assembly in vivo or in vitro. [0151] In summary, self-assembling peptides can be used to create a clean local environment for performing a surgical procedure; isolation of structures and migration of contaminating factors; inflating structures for surgical procedures, i.e. intestines; surrounding structures that are removed that can leak, i.e., the appendix, openings in the body; enabling treatments in dirty environments; they can be used in endoscopic procedures to surround the organ before surgery to stop the leak; used to create a barrier to prevent adhesions during abdominal surgery; and used to create a seal between the endoscopic device and the point of entry of the endoscopic device. Benefits during surgery include the fact that self-assembling peptides are optically clear, have a long service life at room temperature, can be operated through, shorten preparation time, eliminate counting sponges, isolate each structure in the operating field, shorten the cleaning time of the operating room, shorten time of surgery, reduce or eliminate cross contamination caused by other means of washing, the material is biocompatible, the decomposition products are natural and are absorbed by the body. Self-assembling peptides are easily manipulated, they can be injected into the required place, they should eliminate staphylococcal infections, they may be able to reduce the cost of disposable surgical paper, reduce the amount of bags for hazardous biological waste, because the material can be boiled for post-procedure sterilization, obtaining water vapor. Because the self-assembling peptides are clear, this should allow the surgeon to work faster because the surgical field is free of blood. Elimination of the wound tamponade to control bleeding could reduce the duration of action by as much as 50% in a complicated case. Post-operative infection due to secondary infection can be reduced by the use of self-assembling peptides, because they can cover the wound during and after surgery, thereby reducing contamination with foreign bodies. Post-operative care can use self-assembling peptides to reduce infection caused by drainage, slowing the spread of particulate material in the abdominal cavity or chest.

[0152] Although the compositions can be removed at any time from the application site (e.g., the bleeding vessel), the physician may decide that they remain in place even after the initial goal of hemostasis is promoted to promote wound healing. [0153] The compositions comprise self-assembling peptides and these peptides may contain amino acid residues that are naturally occurring and that can be absorbed by the body. The compositions are not difficult to handle and can be easily prepared if necessary. Their properties (e.g. stiffness) can be easily changed by changing the concentrations of their constituents (e.g. by changing the concentration of self-assembling peptides in a given composition). As a result, it has folded the structure does not significantly affect the field of view for the underlying tissue and does not need to be removed before the procedure. For example, a physician may assess a burn or other surface injury that has been treated in the area of ​​the composition described herein. In the operating room, the surgeon can make the initial incision through the material and can continue to work with standard equipment, such as scalpels and clamps or using modern means such as lasers, in the inner area in which the compositions can also be used. Another advantage can be realized over time, because the use of the composition can shorten the time needed to prepare the patient for surgery. Because the compositions can be applied around the incision site and can form a protective coating against infectious agents, there is less need to shave the patient's skin, use curtains and use disinfectants. [0154] Due to the structural integrity of the collapsed scaffolding, it is possible to remove them from the area in which they were created if desired. Thus, a folded scaffold can be removed by, for example, suction or lifting by means of such a device as a tick or by wiping it with a cotton ball or gauze. For example, the scaffold can be removed after haemostasis or during wound cleaning. Based on current research, the scaffolding or its major part can be removed without damage to the tissue underneath. If submerged scaffolding is formed ex vivo, it can be removed from the mold and used later (e.g., implanted into tissue or tissue space). The compositions should reduce the amount of material that needs to be removed or cleaned later (e.g., surgical drapes,

[0155] "Nanosecure" may be used to replace traditional paper or fabric screens, limiting the infection when applied directly to the patient, e.g., by spraying or coating the patient in a different manner or area around the surgical incision. Currently, the patient is preparing for surgery by shaving, cleaning, disinfecting and covering after placing on the operating table. Then, the germicide and the tape are applied to the area in which the surgery is to be performed. The self-assembling composition can be applied instead of curtains by spraying a warm liquid on the body, where it is deposited on a thin second skin. This material has a pore size that is smaller than any bacteria, thanks to that it protects against all contaminants from the air and because a material with a thickness of one millimeter can contain a mild anti-bacterial agent that adheres to the body like the other skin. Self-assembling peptides may also contain a moisturizing ingredient for the skin to not dry out. There is no fear that self-assembling peptides will get to the wound site because they will break down in the body. A bleaching agent can be added, making it easier to determine if the self-assembling peptides have been completely rinsed out after surgery. that self-assembling peptides get into the wound site because they will break down in the body. A bleaching agent can be added, making it easier to determine if the self-assembling peptides have been completely rinsed out after surgery. that self-assembling peptides get into the wound site because they will break down in the body. A bleaching agent can be added, making it easier to determine if the self-assembling peptides have been completely rinsed out after surgery.

[0156] These self-assembling peptides can also be used to prevent the introduction of foreign bodies into the human body and / or the surface of the human body. Self-assembling peptides can prevent the introduction of bacteria, fungi, viruses, spores and / or other infectious agents, creating a barrier that prevents the passage of these materials.

Scaffolding (e.g., a nanoscale structured material) may be provided by introducing a scaffolding precursor into a subject at or in the vicinity of a site where scaffolding is desired (e.g., to control the movement or efflux of a bodily substance to protect the wound or promote repair tissue). Precursors (i.e., self-assembling peptides) are provided adjacent to the site where they are delivered in a place that is sufficiently close to the target area (e.g., bleeding vessel, affected gastrointestinal tract or skin burn area) so that they reach the target area in effective amount. Precursors that may be homogeneous or heterogeneous (e.g. a single type of self-assembling peptide or a mixture of two or more different such peptides may be used), may be included in the composition, and after contact with physiological conditions they assemble to form a scaffold (e.g., nanoscale structured material). Thus, the precursors may be assembled in situ (i.e., in the body of the subject at the site of administration or in its vicinity).

[0158] The nanoscale structured material may or may have a complex structure and may contain additional components present in situ (e.g., ions). Thus, precursors, such as self-assembling peptides, can be used in a substantially ion-free solution (e.g., substantially free of monovalent cations) and can self-assemble, forming a macroscopic structure after they come into contact with such ions in the body (e.g. in a systemic substance such as blood, digestive tract content and the like). For example, a solution containing precursors can be used in or adjacent to the perforation site of the stomach or intestine, or the location where surgical cleavage has been or will be performed.

[0159] The scaffold may also be provided as a gel, because precursors (i.e., self-assembling peptides) can be assembled before the composition is introduced into the target area (e.g., the site where the incision will be made for the surgical procedure). The folded structure can take any convenient shape.

[0160] Scaffolding can also be provided by providing precursors in the form of a dry powder. The "dry" powder will have a relatively low liquid content (e.g., sufficiently low that the particles in it are easily disperseable). The self-assembling peptides provided in the form of a dry powder will collapse after they come into contact with a body fluid containing monovalent cations, and a solution containing such ions can be added, if desired, to change the speed at which the scaffold is formed or stiff. Self-assembling peptides may be provided as emulsions or, as described above, formed into pre-formed shapes that can be inserted into the body cavity or wound site in a manner similar to that in which surgical sponges are currently used. If necessary, a binder can be added to the dry powder, which is then formed into the desired shape. Regardless of the exact method of assembling the scaffold (e.g., by contacting a liquid preparation containing precursors with the body or a dry powder with an ex vivo ion-containing solution), the scaffolds formed may take the desired shape. In the case where the size and shape are such that the scaffolding fills the lumen of the blood vessel, the scaffold can be used as a vascular plug.

[0161] A preventive action can be carried out before the subject has an undesired event (e.g., prior to the onset of injury or before the onset of bleeding). Thus, the place of administration may be a place of potential movement or potential leakage and, by application, the displacement or leakage may be prevented or minimized by application. In the context of a therapeutic procedure or treatment, the compositions may reverse, alleviate or suppress the condition (e.g., disease, syndrome, disease or symptom, subjective symptom or sign). The methods of treating the subject are generally carried out after the subject has been recognized as having a condition treatable, and any of the methods described herein, whether best described as prophylactic or therapeutic,

[0162] Since the compositions described herein can be used to inhibit the movement of a body substance in a subject, including intra- or epidermal movement, the compositions can be used in the context of a surgical procedure and can be described as new methods for performing a surgical operation or for creating an operating field. The methods, whether performed in the context of a cancer surgery or not, may include the step of identifying the subject in need of treatment and the step of providing the nanoscale structured material or precursor thereof at or in the vicinity of where unwanted movement has occurred or is about to occur. For example, you can identify a patient, which is to undergo a surgical procedure and which is to be provided with a biocompatible composition comprising self-assembling peptides and a vasoconstrictor, colorant or local anesthetic to the location where the incision or other invasive maneuver will be performed or performed. The systemic substance concerned may be a liquid such as blood or blood product, a serum exudate (effusion associated with inflammation, consisting mainly of plasma that usually appears as a transparent or colored amber liquid), pus, gastric juice , urine, bile, cerebrospinal fluid (CSF), pancreatic juice, and the like. A constituent substance may be viscous, sludgy or semi-solid, but generally exhibits the ability to flow or move. Substances of this type include the content of the gastrointestinal tract. The composition can be removed after application (e.g. after hemostasis or end of intestinal surgery) or it can be left in place. For example, the compositions can be used to accelerate hemostasis or inhibit the dislocation of intestinal contents during surgery, and some or all of the scaffold can be left in place when the operation is completed. This provides a significant advantage over the use of sponges and other materials that must be removed before closing. The compositions can be removed in various ways (e.g., by wiping or aspiration). For example, the compositions can be used to accelerate hemostasis or inhibit the dislocation of intestinal contents during surgery, and some or all of the scaffold can be left in place when the operation is completed. This provides a significant advantage over the use of sponges and other materials that must be removed before closing. The compositions can be removed in various ways (e.g., by wiping or aspiration). For example, the compositions can be used to accelerate hemostasis or inhibit the dislocation of intestinal contents during surgery, and some or all of the scaffold can be left in place when the operation is completed. This provides a significant advantage over the use of sponges and other materials that must be removed before closing. The compositions can be removed in various ways (e.g., by wiping or aspiration).

[0163] The compositions may also be used to shield the underlying area (e.g., the area burned or otherwise damaged wound or other tissue), and thus may help to prevent contact of contaminants (e.g., foreign matter) with the area (i.e. the compositions can be used as a barrier or shield). A doctor or other healthcare provider can examine the wound through self-assembling peptides, and the surgeon can operate through them while they are in place. Contaminants that have found themselves on self-assembling peptides during the procedure can then be removed by removing the peptides.

[0164] The compositions may be administered to stabilize the wound prior to final treatment (e.g., while the victim is awaiting transport to hospital or during transport). The compositions are similarly useful when operations are carried out under conditions of less than optimal sterility (e.g., in field hospitals or in places in the world where access to sterile operating rooms is limited). Compositions and methods can potentially significantly reduce the likelihood of contamination in such cases.

[0165] Self-assembling peptides may also be applied locally in combination with an anesthetic to a local area where the procedure is to be performed and may be used at a higher concentration to reduce organ displacement during surgery. This can reduce cognitive deficits in older patients by reducing the overall anesthetic burden. A thin layer can be sprayed onto the tissue or skin that the surgeon uses. They can be used separately or together, giving a specific anesthetic for specific organs. The skin has different receptors than the intestines and there is a need for a special anesthetic for each of the organs. The intestines must stop moving during the surgery, while the blood and spasm of blood vessels must remain unchanged.

[0166] Treatment and Prevention of Bleeding: Any subject at an increased risk of undesirable bleeding that may or may not be excessive or imminent life threatening may be treated with the compositions described herein. These individuals include people with blood coagulation disorders such as haemophilia, patients receiving anti-coagulant therapy, patients suffering from recurrent nosebleeds and subjects undergoing surgery, in particular major surgery or procedures that require access to the artery.

Unbound, a surgical procedure or procedure may be an operation within the nervous system, eye, ear, nose, mouth, throat, respiratory system, cardiovascular system, digestive system, urinary tract, musculoskeletal system, skin system or system reproductive. Specific examples of surgical procedures and procedures in which the compositions may be used include arteriography, angiocardiography, cardiac catheterization, repair of obstetric lesions, removal of coronary artery occlusion, stent insertion, caesarean section, hysterectomy, fracture bias, coronary artery bypass surgery, cholecystectomy, organ transplantation , replacement of the entire joint (eg knees, hips, ankles, shoulder), apendectomy, excision or destruction of the intervertebral disc, partial excision of the large intestine,

[0167] Accident victims, individuals involved in fighting, and women who are giving birth are also exposed to significant blood loss. The compositions may be applied to the place of obstetric bleeding (e.g., inside the uterus, vagina or adjacent tissue) to accelerate hemostasis. For example, the compositions can be applied to the tear of the placenta or applied to the uterine tamponade to control bleeding. As with other indications, compositions applied to reproductive routes can be removed or left in place. Spontaneous haemorrhage, aneurysm rupture, esophageal varices, stomach ulcers, upper intestinal ulcers (e.g. duodenal ulcers) are also medical conditions in which significant bleeding can occur and these individuals can also be treated as described herein.

[0168] The exact source of bleeding may vary and may come from blood vessels in the arterial or venous system (e.g., artery, arterioles, capillaries, capillary beds, veins or veins). The size of the vessel may vary from large (e.g., compositions may inhibit bleeding from the aorta, iliac or femoral artery or portal vein) to a small one (e.g., capillaries), and the vessels can be located anywhere in the body (eg in a solid organ such as like liver, stomach, intestine, skin, muscle, bone, lungs or reproductive system).

[0169] The time normally required for blood clotting may be prolonged when the levels of coagulation factors and / or platelets in the plasma are low or in cases in which the subject received an anticoagulant (e.g., warfarin or heparin). Bleeding often lasts for much longer than the average clotting time when there is more than minimal damage to the integrity of the blood vessels. On the basis of the studies, the compositions are expected to cause haemostasis in less than or at least in some cases much shorter than the average blood coagulation time. Although the compositions are not limited to those that achieve haemostasis at a given time (and uses, such as protecting the area from contamination or promoting tissue healing, are independent of this function), the compositions can benefit the bleeding subject in such a short time as five seconds after application. Other compositions may exert their effect within about 10, 15 or 20 seconds after application. An effective period can be characterized in a different way than an absolute one. For example, the compositions may reduce the time required to achieve hemostasis between 25% and 50%; between 50% and 75%; or between 75% and 100% in relation to the time required when using ice cold saline. The time required to achieve haemostasis can be reduced by approximately 2-, 3-, 4- or 5-fold relative to the time required when using ice cold saline. An effective period can be characterized in a different way than an absolute one. For example, the compositions may reduce the time required to achieve hemostasis between 25% and 50%; between 50% and 75%; or between 75% and 100% in relation to the time required when using ice cold saline. The time required to achieve haemostasis can be reduced by approximately 2-, 3-, 4- or 5-fold relative to the time required when using ice cold saline. An effective period can be characterized in a different way than an absolute one. For example, the compositions may reduce the time required to achieve hemostasis between 25% and 50%; between 50% and 75%; or between 75% and 100% in relation to the time required when using ice cold saline. The time required to achieve haemostasis can be reduced by approximately 2-, 3-, 4- or 5-fold relative to the time required when using ice cold saline.

[0170] The concentration of self-assembling peptides can be selected with reference to variables such as the size of the vessel, the extent to which it was damaged, and the force with which the blood escapes (or could escape after being wounded). Higher concentrations of peptides will be desirable to promote hemostasis from the main vessel (e.g., aorta, brachiocephalic artery, carotid, subclavian artery, celiac trunk, mesenteric, renal, iliac, femoral or popliteal). Useful concentrations may range from between about 0.1-10% (e.g., 1-10%, 0.5-5%, 1-4%, 0.1-2%, 0.1-3%; 1-4%, 0.1-5%, and 1-8% (e.g., about 1, 1.5, 2, 2.5, 3, 4, 5, 6 or 7%). Any sub-range or any particular value from any of the aforementioned ranges.Any of the aforementioned concentrations may also be used in the other indications described herein.

[0171] As mentioned, the bleeding may be due to any of a variety of different reasons and may be internal or external. The compositions can be applied regardless of the cause or nature of the cause (e.g. whether it is due to a disease process or an intentional or accidental injury). The compositions may be used to achieve haemostasis in a confined space (e.g., inside an hollow organ) or on or near the body surface. For example, the compositions may be applied to a partially or fully separated part of the body, such as a limb or finger. In this case, the compositions can be fullmany functions; they can not only promote hemostasis, but also protect damaged tissue from contaminating agents and promote tissue healing. More specifically, the compositions can be applied to the wound, left in place for a period of time sufficient to achieve haemostasis and blood clotting, and then removed. Contaminant material, such as solid particles and infectious agents adhering to the peptide gel, can be removed. A sterile dressing can then be used. Of course, the compositions can be applied to purify the wound, prevent contamination or promote tissue healing even after haemostasis or in situations in which acceleration of hemostasis is not needed.

[0172] When used for the treatment of epistaxis, the compositions are incorporated into a suitable nostril and may remain in place until the bleeding is resolved. The compositions can be easily removed by suction (e.g. with a dropper or syringe) or can be removed with other physical means, including simply blowing the nose.

[0173] The compositions may also be left in place of the wound and a dressing may be applied to the composition. Because the composition itself is easily removed, its presence under the dressing can prevent the dressing from adhering to the damaged tissue. If desired, a bandage with a transparent part can be used so that the damaged area can be viewed through the transparent part of the bandage and the peptide structure below. This will allow the doctor to monitor the progress of healing without removing the dressing. Modified bandages are described below and are within the scope of the present invention.

[0174] Many medical procedures involve puncture of the vessels that may be accompanied by significant bleeding. The composition of self-assembling peptides can be applied to the wall of a pierced vessel, e.g. when withdrawing the device used for puncturing the vessel. The vascular plug formed of self-assembling peptides is an alternative to existing vascular plugs and devices, such as those described in US Patent No. 5,192,302; 5222974; 5645565; and 6663655. The vascular plug may be formed in situ (e.g., at the point of puncture of the vessels) or it may be pre-formed and used in place.

[0175] More generally, compositions comprising nanostructured materials or their precursors (i.e., self-assembling peptides) can be used to seal any tissue passage. The present methods thus include methods for sealing a tissue passage by applying a composition comprising a nanoscale structured material (e.g., self-assembling amphiphilic peptides) to one or both of the transition ends or interiors thereof. The tissue may be, for example, the wall of a blood vessel, an organ wall, subcutaneous tissue or adipose tissue. Sealing the passage may cause hemostasis. The transition may also be a fistula (ie an abnormal connection between two organs or the structure of the organism or between the organ or structure and the outside world). If necessary, the surgeon may apply the compositions to the inside of the tubular structure, such as the intestine or blood vessel, to cut and connect the gut or blood vessels in the gel, and then to remove the gel from the inside of the structure to restore the structure and allow the area to reperfusion with blood or other body substances. Self-assembling peptides can also be used to limit reperfusion injury. For example, self-assembling peptides may be administered after ischemia, such as in patients treated with a thrombolytic agent. Self-assembling peptides can also be used to limit reperfusion injury by restoring blood tissue barriers before, during and / or after reperfusion. For example, self-assembling peptides can be used to restore the tissue-blood barrier through the inner shell of a portion of the circulatory system. This may be beneficial in diseases such as ischemic infarction, haemorrhagic stroke or reperfusion injury. Finally, self-assembling peptides can be used to limit reperfusion injury by restoring the integrity of the vascular structure before, during and / or after reperfusion.

[0176] For surgical applications, a wound or any portion of the operative field may be padded using a composition containing self-depositing peptides. This approach can be used instead of wound tamponade, because it is conventionally performed during surgery. Because the compositions contain biocompatible and biodegradable material, they can be left in place, avoiding the need to remove after the procedure and avoiding the need for subsequent surgery for this purpose. Biodegradable materials can be broken down physically and / or chemically in the cells or in the body of the subject (e.g., by hydrolysis under physiological conditions or through natural biological processes, such as the action of enzymes present in the cells or in the body), with the formation of smaller chemical molecules that can be metabolized and, optionally, re-used and / or excreted or otherwise removed. Preferably, biodegradable compounds are biocompatible.

[0177] Gastrointestinal bleeding that can occur as a result of ulcers or angiodysplasias is a relatively common and serious condition that can be fatal if left untreated. Bleeding of esophageal varices and bleeding of stomach ulcers or duodenal ulcers can be particularly severe. Numerous endoscopic therapeutic approaches have been developed to achieve haemostasis, such as the injection of sclerosing agents, the attachment of mechanical hemostatic devices, and contact electrocautery techniques. The compositions may be administered to or in the vicinity of an ulcer or at the site of bleeding in the esophagus, stomach, small intestine or large intestine.

Bleeding in the distal part of the large intestine, rectum or anus (eg haemorrhoids) can also be treated in this way.

[0178] An aneurysm rupture can be a catastrophic event with rapid fatal consequences. Ruptured aortic aneurysms can quickly lead to bleeding after 5 rapid medical interventions. Broken intracranial aneurysms often have devastating consequences. The compositions and methods of the invention can be used to treat bleeding from a ruptured aneurysm in a manner substantially similar to the way they are used to treat bleeding for other reasons (e.g., by using self-proprietary precursors or a pre-formed structure at a bleeding site). Due to the often serious consequences of aneurysm rupture, it is often attempted to repair it surgically. The compositions can be used in the context of any repairs (e.g. during open surgery or intravascular repair (eg with placement of a graft and / or stent)). More specifically, the present methods comprise treating an aneurysm by introducing a composition comprising a nekaline structured material or a precursor thereof (e.g., a composition comprising self-assembling peptides) into an aneurysm (e.g., an aneurysm sac). When any bleeding is under control, the aneurysm can then be repaired using any suitable technique. The presence of a peptide structure in the aneurysm sac reduces the risk of leakage or rupture before or during other procedures. The scaffolding can be left in place. In addition, the presence of material in the aneurysm sac can promote healing of the aneurysm. More specifically, the present methods comprise treating an aneurysm by introducing a composition comprising a nekaline structured material or a precursor thereof (e.g., a composition comprising self-assembling peptides) into an aneurysm (e.g., an aneurysm sac). When any bleeding is under control, the aneurysm can then be repaired using any suitable technique. The presence of a peptide structure in the aneurysm sac reduces the risk of leakage or rupture before or during other procedures. The scaffolding can be left in place. In addition, the presence of material in the aneurysm sac can promote healing of the aneurysm. More specifically, the present methods comprise treating an aneurysm by introducing a composition comprising a nekaline structured material or a precursor thereof (e.g., a composition comprising self-assembling peptides) into an aneurysm (e.g., an aneurysm sac). When any bleeding is under control, the aneurysm can then be repaired using any suitable technique. The presence of a peptide structure in the aneurysm sac reduces the risk of leakage or rupture before or during other procedures. The scaffolding can be left in place. In addition, the presence of material in the aneurysm sac can promote healing of the aneurysm. When any bleeding is under control, the aneurysm can then be repaired using any suitable technique. The presence of a peptide structure in the aneurysm sac reduces the risk of leakage or rupture before or during other procedures. The scaffolding can be left in place. In addition, the presence of material in the aneurysm sac can promote healing of the aneurysm. When any bleeding is under control, the aneurysm can then be repaired using any suitable technique. The presence of a peptide structure in the aneurysm sac reduces the risk of leakage or rupture before or during other procedures. The scaffolding can be left in place. In addition, the presence of material in the aneurysm sac can promote healing of the aneurysm.

[0179] Inhibition of cerebrospinal fluid (CSF) movement or leakage: The hard tire is a hard, outermost, fibrous membrane that covers the brain and spinal cord and defines the inner surface of the skull. CSF leakage is a significant complication following wounding, surgery or other procedure in which the dura mater is penetrated, including accidental penetration during administration of the anesthetic into the epidural space. Such a leak can lead to serious consequences, such as severe headaches, infection and meningitis. The composition may inhibit displacement or CSF leakage in a subject in need thereof after application at or in the vicinity of an undesired CSF movement or leakage site. The compositions may be applied to the sutures after dural surgery to prevent CSF leakage from the incision site. The compositions may also be used to inhibit the movement or leakage of fluid from the eardrum.

[0180] Inhibition of gastrointestinal tract leakage: The compositions may inhibit the dislocation of digestive tract content. For example, structures can prevent leakage of gastrointestinal contents after gastric or intestinal perforation, or during surgery (see Example 4). The structures can be used to isolate such body substances and prevent their spreading in the peritoneal cavity, thus minimizing contamination and the risk of chemical peritonitis and / or infection. The stomach contents that contain digestive secretions of the stomach glands, consisting mainly of hydrochloric acid, mucin and enzymes such as pepsin and lipase, can cause injury and / or infection if released into the peritoneal cavity. The release of intestinal contents into the peritoneal cavity is a frequent event during intestinal surgery and may also occur in the case of intestinal perforation or ruptured appendage. The composition can be used to inhibit the leakage of gastrointestinal contents into the peritoneal cavity. The place of movement can be a place of damage to the stomach or intestines caused by the disease process or surgical incision. The compositions can be applied to the outside of any organ in the digestive system (e.g., stomach or small or large intestine) or they can be injected or otherwise injected into their interior. The compositions may be administered during resection of the intestinal segment. For example, a section of the intestine can be filled, which extends from the first point to the second point with the present composition, and part of the intestine is cut out, which lies between the first and the second point. In one embodiment, self-assembling peptides may be used to treat heartburn. For example, self-assembling peptides may be formulated as a solution, suspension or emulsion (such as a beverage or cocktail), a gel, a tablet, a cachet, a capsule, etc., administered orally to cover part of the gastrointestinal tract. The preparations can be used to: stop the movement of body fluids, including gastric juices and blood; coating of the JM conductor and / or stopping ulcer progress, erosion and inflammation. The preparations can be used to prevent damage to the esophagus due to reflux disease. The formulations may also be used to assist in the repair of cells in the esophagus that have been damaged by acid reflux, other diseases or disorders and / or therapeutic interventions. The preparations can be used to help repair primary and secondary ulcers and erosion in the mucous membrane. The formulations may be used to deliver therapeutic, prophylactic and / or diagnostic agents to a portion of the GI junction as necessary. For example, self-assembling peptides may be used to provide means for re-establishing the flora and fauna of the GI tract that have been removed by radiotherapy and / or disease or injury.

[0181] In the associated method, compositions for removing intestinal contents that have been released into the peritoneal cavity can be used. The method includes applying a liquid composition to the released intestinal contents, allowing the liquid phase transition composition, and then removing the gel-like or semi-solid composition. These steps can be repeated once or more until the surgeon is satisfied with the amount of intestinal contents that have been removed from the peritoneal cavity. In another related method, the compositions may be applied to ulcers to act as a barrier to prevent contact of the acid with the stomach surface.

[0182] Likewise, the dislocation of the contents of other internal organs (e.g., organs in the bile or urinary system) can be inhibited. For example, the movement of bile, pancreatic juice (ie exudate exudates of the pancreas that contain digestive enzymes) or urine and / or disinfect or purify an area in which bile, pancreatic juice or urine is released by applying, and then removing the composition from it. These methods are widely used in surgical procedures to repair or otherwise treat intestinal, biliary and / or urinary defects.

[0183] Wound healing: Studies also show that the compositions have the ability to enhance healing, particularly of the epithelial or muscle layer, and thus can be administered to treat the site of tissue damage. For example, a composition comprising self-assembling peptides at the site of tissue injury may be used. It turns out that the compositions increase the rate of tissue repair and inhibit the formation of scar tissue. The compositions can be used for the care of acute or chronic wounds. For example, they can be applied to the skin injured in any way (e.g., torn or burned) and for lesions such as ulcers and pressure sores. In the case of burns, the self-assembling composition, optionally containing a cleansing agent, can be administered to the site of the burn. Purification could take place in or through self-assembling peptides, thereby reducing the degree of abrasion in place, minimizing or eliminating contamination of the surrounding tissue or environment. As a result of the assembly of self-assembling peptides, a barrier is also formed that can prevent infection and / or contamination of the burn.

[0184] Self-assembling peptides may also be useful for inhibiting or preventing the formation of scar tissue by chelating iron and other metal ions that act as cofactors for the formation of scar tissue. When the wound heals, the pH of the wound site drops and the iron concentration increases. Scarring and scar tissue may block axon growth. Self-assembling peptides can block the formation of scar tissue by chelating iron at the wound site, presumably through the presence of electronegative and / or negatively charged functional groups that can complex positively charged metal ions. Removal or complexation of iron in the wound prevents the stable formation of collagen IV. The ability to control the wound environment allows you to control the rate and degree of healing.

[0185] This ability of self-assembling peptides to chelate metal ions may also be useful in preventing bacterial or fungal infections by chelating metals that are cofactors for bacteria and fungi. Infections can also be prevented by coating the tissue with a layer of self-assembling peptides, thereby preventing the attachment of bacteria or fungi to the tissue.

[0186] These self-assembling peptides can be used to maintain the hydration and nutrition of patients who have had burns or in cases where the outer skin has been compromised due to abrasion or burns.

[0187] In another case, self-assembling peptides can be used to maintain body temperature when the patient is coated with peptides by a source of external heat or cooling.

Tissue Regeneration [0188] Vehicle for drug delivery to the intrathecal space: The self-assembling peptides described can be used to deliver therapeutic and / or imaging agents to the intrathecal space. Examples of therapeutic agents include, but are not limited to, anti-inflammatory agents and nerve / spinal cord stimulation stimulants. A hydrogel material was used in an attempt to deliver one or more active agents to the intrathecal space. However, these materials can be limited by their slow polymerization times, which allows diffusion of the material after the material polymerizes to form a gel. The self-assembling peptides described can be designed for rapid self-assembly so that these peptides do not diffuse.

[0189] Cartilage Repair: The self-assembling peptides described herein can be used to repair cartilage. Self-assembling peptides would typically be injected into the place where cartilage repair is required. Self-assembling peptides can be used alone or in combination with cells and / or growth factors.

[0190] Bone regeneration: Self-assembling peptides described herein can be used to prepare composite materials for bone regeneration. For example, self-assembling peptides may act as a carrier for inorganic materials such as calcium phosphate or hydroxyapatite, organic materials such as growth factors and / or bone grafts. Inorganic materials, such as calcium phosphate, can be remodeled using the osteoclast resorption mechanism for bone regeneration. Self-assembling peptides can be injected under periosteum to stimulate bone growth as agents for the formation of bone grafts in vivo. Alternatively, the self-assembling peptides described can be used to carry out bone regeneration therapies that limit fibrous growth. For example,

[0191] Oxygen Supply: Self-assembling peptides described herein can also be used to deliver oxygen to the lungs and / or other organs. For example, self-assembling peptides can be oxidized to a very high degree to provide oxygen / perfusion in patients suffering from pulmonary haemorrhage and other lung diseases.

[0192] Methods, devices and delivery kits: Many devices may be used to introduce the composition to the target area of ​​the organism. The devices can be simple, such as a syringe and such devices can be supplied with the compositions in sets. The composition may be delivered locally at or near the target area in the body by injection (e.g., using a needle and syringe) or a catheter, cannula or by dispensing (e.g., casting) from any vessel of appropriate size. The compositions may be delivered using imaging guidelines (e.g., stereotaxic instructions), if necessary. Alternatively, the material may be wetted with the composition and then used to apply the composition to the tissue area.

[0193] For storage and transport, the self-assembling peptides can be dissolved in a suitable solvent (e.g., an aqueous medium such as sterile water and stored for a longer period of time prior to use). Solutions containing the peptide were stored for up to two years without significant loss of activity. If there is partial self-assembly after a longer period, physical mixing (e.g.

sonication) to restore self-assembling peptides to a more fluid state prior to administration. Alternatively, self-assembling peptides can be used as a gel. If desired, a small amount of ions (e.g., monovalent cations) can be added to the solution before use. This can speed up the gel creation process. Alternative5 monovalent cations can not be used after administration of the solution.

[0194] Kits comprising syringes with different capacities are provided or vessels with deformable sides (e.g., plastic vessels or plastic side dishes) that can be compressed to squeeze the liquid composition out of the opening. In one embodiment, the syringe or vessel comprises a plurality of compartments, one containing monovalent ions and other self-assembling peptides that are mixed during administration by a common needle. The endoscope can be used to deliver compositions for the treatment of hollow organ (e.g. esophagus, stomach, intestine, etc.) or the body cavity (e.g., during minimally invasive surgical procedures). A minimally invasive surgical procedure refers to the way a surgical procedure is performed, in which the operations are carried out with the use of specialized instruments intended to be inserted through small incisions or natural openings of the body, often performed with endoscopic visualization. Examples include laparoscopic surgery, arthroscopic surgery and vascular surgery. The endoscope is usually a long, flexible device similar to a tube. In addition to allowing the visualization of internal structures, many endoscopes have additional diagnostic (e.g., biopsy) and therapeutic (e.g., delivery of therapeutic agents) possibilities via special channels. Colonoscopes, sigmoidoscopes, bronchoscopes, cystoscopes and laparoscopes are variants of the endoscope that exhibit particularly useful features for viewing certain organs, structures or cavities. Any of these devices may be used to deliver the composition. Kits may be packaged, including an endoscope and a vessel containing a solution containing self-assembling peptides. Suitable endoscopes are known in the art and are widely available. Endoscopes are currently used to deliver sclerosing agents to the site of esophageal bleeding.

[0195] Kits may include self-assembling peptides and one or more of: a syringe, needle, thread, gauze, bandage, disinfectant, antibiotic, local anesthetic, analgesic, surgical suture, scissors, scalpel, sterile liquid and a sterile vessel . The peptides may be in solution or in a dry state (e.g., in the form of a dry powder). The components of the kit can be packed individually and are sterile. Kits are usually provided in a container, e.g. a plastic, cardboard or metal container suitable for sale in commerce. The kit can be stylized as a "first aid kit", in which case it will usually have a symbol, such as a red cross outside. Each kit may contain instructions for use.

Examples

Example 1: Material from self-assembling peptides accelerates hemostasis in the brain [0196] Full branch cleavage of the sagittal sinus was performed in the brains of rats and hamsters after removal of the part of the skull covering the tissue undergoing cutting. The animals were anesthetized by intraperitoneal (ip) injection of ketamine (80 mg / kg) and xylazine (8 mg / kg). All surgical procedures were performed under an operating microscope. Twenty-two animals, including 10 adult hamsters and 12 young female Spraque-Dawley rats (200-250 g), treated with ice cold saline or 20 μl of a 1% peptide solution at the cut point of the sinus branch. The material was prepared by dissolving the RADA16-1 peptide (n-RADARADARADARADA-c; SEQ ID NO: 1) in fresh water and the solution containing the peptide was applied to the damaged tissue with a 31G needle attached to 2 cm³ syringes .

[0197] The experiment was recorded with a time stamp and recreated frame by frame to assess the time it takes to form a gel through a peptide solution that effectively inhibited bleeding. Hemostasis was assessed visually, and "total hemostasis" was defined as the total lack of blood flow from the wound site. Complete haemostasis was achieved within 10 seconds after application of the peptide solution in all cases.

[0198] A series of pictures of an adult rat were taken, at which part of the covering skull was removed and one of the veins of the superior sagittal sinus was incised and then treated with a solution containing the peptide. The first picture shows the exposed brain and veins of the sagittal sinus; the next picture shows the cut of a vein; the next picture shows the bleeding from a broken vein; and the final image shows the same area five seconds after applying the peptide solution. Complete hemostasis was obtained.

[0199] Periods were measured from the start of application of the peptide solution to completion of haemostasis after cleavage of veins leading to the sinus in the brains of adult rats. Total hemostasis is achieved on average in 8.3 seconds. In physiological saline control, bleeding was never achieved. The control experiment with saline was discontinued at the same time point to prevent the animals from bleeding out to death.

[0200] Similar results were obtained after complete sagittal sinus cleavage. In this second experiment a higher concentration of peptide (e.g., about 3% - 4%) was used to achieve hemostasis. Three controls with saline continued to bleed after 20 seconds. In the case of control animals, ice-cold saline was removed and a peptide solution was applied, which resulted in complete haemostasis almost immediately.

[0201] A total of 22 rats and 64 hamsters were subjected to experiments in which solutions containing the peptide effectively allowed to achieve haemostasis within 10 seconds after administration to the site of intracranial hemorrhage.

Example 2: Material from self-assembling peptides accelerates haemostasis after cleaving of the femoral artery [0202] A sciatic nerve and adjacent femoral artery were exposed to adult rats and the femoral artery was incision. Twelve rats were treated by applying μΐ of a 1% RADA16-1 peptide solution (SEQ ID NO: 1) to the cleavage site using a glass pipette attached to the syringe body, while controls were treated by applying cold saline to the cleavage site. In all treated cases, hemostasis was achieved in less than 10 seconds. Control cases with saline continued to bleed until the end of the experiment after 110 seconds. In these control animals,

[0203] A series of pictures of an adult rat with a femoral artery was made. In the first picture, the sciatic nerve and femoral artery are exposed. The next picture shows the artery cut, and the next picture shows the bleeding. After about five seconds, complete hemostasis was observed in the region of the clear gel formed by the deposition of peptides in the presence of blood and plasma. The deposited material can be easily sucked out of the place, if desired. Complete haemostasis was maintained for the duration of the study (1 hour).

[0204] Complete haemostasis was achieved in less than 10 seconds. Haemostasis has never been achieved with saline control.

[0205] Experiments with muscle injury showed immediate haemostasis after 1-2 cm incisions of the dorsal muscle of the rat. Quadrilateral muscles were exposed on the back of rats and a deep muscle incision was made, followed by applying a 1% peptide solution (RADA16-I) to the incision (SEQ ID NO: 1). Within 10 seconds, all the bleeding stopped. In the case of ice-cold saline application, the control animals continued to bleed after 20 seconds.

[0206] This procedure was repeated in the hind limb muscle (porteocaudalis and musculus tibialis cranialis) and similar results were obtained. Between 1% and 100% peptide (RADA16-1) (SEQ ID NO: 1) was applied to limb wounds and haemostasis was found in all cases. However, in the case of an arterial or vein incision, 2% or more material was needed to bring about haemostasis. In the case of ice-cold saline application, the control animals continued to bleed after 20 seconds.

Example 3: Material from self-assembling peptides accelerates hemostasis in the liver [0207] In order to further demonstrate the ability of structures containing the peptide to inhibit bleeding of a relatively low pressure vessel, an adult rat's peritoneal cavity was opened, the liver was exposed, and the sinner lateralis was incision dorsolateral by completely crossing part of the liver. There was profuse bleeding. A 1% peptide solution (RADA16-1) (SEQ ID NO: 1) was applied to the cut and close to it using a 27G needle and 4cm³ syringes. All bleeds stopped within 10 seconds. A series of photos were obtained. The first one shows the exposure of the liver; on the second, the liver is separated, and heavy bleeding is visible; and on the third, two parts of the liver can reconnect and the bleeding continues. After treating the site with a 1% peptide solution (applied locally and at the cut site), all bleeding resolved within 10 seconds. A clear area was observed between the two parts of the sinus lateralis. This procedure was repeated several times with the same effect.

[0208] A similar experiment showed the ability of peptide structures to stop vessel bleeding in a higher pressure liver. The experience is illustrated by a series of photos. The first one shows the open peritoneal cavity and the exposed liver; on the second lobus sinister lateralis received a transverse cut completely crossing the part of the liver and the main branch of the portal vein; and the third shows heavy bleeding from the wound. The cut was treated with a 4% peptide solution applied topically and in a cut. All the bleeding resolved within 10 seconds. The lower part of the lobus sinister lateralis was pulled down to show that the peptide structure is in the cut. The place did not bleed even when subjected to this physical stress Ten minutes later, there was still no bleeding. Thus,

[0209] Treatment with a 2% or 3% peptide solution was tested in the same type of experiment and complete haemostasis was also achieved. Treatment with a 1% solution resulted in partial cessation of bleeding. Furthermore, 30 seconds after the treatment, the excess peptide structure was removed from the wound site and hemostasis was maintained.

This procedure was repeated several times with the same effect.

[0210] In other experiments, one quarter of the lobe was removed in the lower right quadrant of lobus sinistras laterialis, and the rim was treated by topical application of a 2% peptide solution (RADA16-I) (SEQ ID NO: 1) at the wound site. Bleeding stopped in less than 10 seconds. After one minute the peptide was removed and complete hemostasis was achieved on the periphery of the liver.

Example 4: Material from self-assembling peptides to prevent adhesions Liver in 18 adult rats were exposed in deep general anesthesia, an opening was made in the upper right panel with 4 mm of trepane, and then the wound was treated with 3% NHS-1. The animals were allowed to survive 2 days, 7 days, 14 days, 6 weeks, and 8 weeks respectively, the animals were again anesthetized and the lobe of the liver with the hole cut was prepared and processed for H &amp; E staining. In addition, the control set was treated with saline or cauter.

[0212] Experiment with a 4 mm biopsy of liver liver, with control and filling of the cut hole with 3% RADA16-I solution (SEQ ID NO: 1). All controls were green on both surfaces, while those treated had no adhesions. Within 2 weeks, weeks and 8 weeks, control adhesions on the upper and lower surfaces were excised. For all cases treated with a 3% RADA16-I solution (SEQ ID NO: 1), no adhesions were noted on the upper or lower surface of the liver.

Example 5: Material from self-assembling peptides [0213] Adult rat was perforated with a small incision at the level of the duodenum, which caused leakage of gastric fluid into the peritoneal cavity. When the site was treated with a 2% peptide solution (RADA16-I) (SEQ ID NO: 1), all leakage of gastric fluids from the gut ceased. An additional volume of 2% peptide solution was injected into the duodenum at the wound level. This prevented leakage from the intestine for one hour, the duration of the procedure. During the control cut at the duodenum level, the intestinal wall dislocated and the gastric fluid continued to leak from the wound site in the case of no treatment. When the site was treated with the peptide solution 15 minutes after wounding, the peptide treatment also stopped all spills from this wound site. Also,

Example 6: Material from self-accumulating peptides accelerates skin wound healing [0214] To demonstrate the ability of self-assembling peptides to enhance wound healing, animals were biopsied with skin and subcutaneous trephine. The areas in which the biopsies were performed were treated by single application of a self-assembling peptide solution (RADA16-I) (SEQ ID NO: 1) or other untreated. The wounds remained un-massed. The results illustrate a series of pictures from the healing test after a 4 mm biopsy with trepane, in which injured animals were treated with a self-assembling peptide and compared with the corresponding untreated cases. The wounds were photographed on day 0, day 1, day 4 and day 7. The wounds healed were healed much faster, which was confirmed by narrowing the wound site in all three cut holes on day 1. It turns out that peptide treatment has accelerated healing by up to 5 days in some cases. In all cases, contraction of the wound site occurred faster in the treated cases.

Example 7: Lidocaine-containing compositions [0215] RADA16-I (SEQ ID NO: 1) was mixed with lidocaine and the mixture was applied to the skin of adult rats prior to application of the pin with a pin. It is a 5% blend of lidocaine and RADA16-I (SEQ ID NO: 1). Applied on the skin and left for the duration of the study. After mixing with the self-assembled peptide, the pin response was silenced four times longer than the response, which was silenced using lidocaine alone. In addition, solutions of self-assembling peptides and lidocaine were applied to the intestines of two rats during intestinal surgery. The solution reduced the peristalsis throughout the entire surgical procedure without visible side effects in the animals.

The following paragraphs provide a general description of the invention.

A method for preventing or reducing adhesions comprising administering to a demanding site, in the absence or substantial arrest of bleeding or fluid leakage, of a self-assembling material that creates a barrier to or otherwise prevents the formation of adhesions.

2. The method of paragraph 1, wherein the self-assembling material comprises peptides having a sequence of amino acid residues corresponding to one or more of formulas I-IV:

<td>((Xaa^neu-Xaa⁺) X (Xaa^neu -Xaa^-) Y) n</td><td>(AND)</td>

<td>((Xaa^neu -Xaa^-) X (Xaa^neu-Xaa⁺) Y) n</td><td>(II)</td>

<td>((Xaa⁺-Xaa^neu) X (Xaa^--Xaa^neu) Y) n</td><td>(III)</td>

((Xaa^--Xaa^neu) X (Xaa⁺-Xaa^neu) y) n (IV) wherein Xaa^neu means an amino acid residue having an inert charge; Xaa⁺ is an amino acid residue having a positive charge; Xaa^- is an amino acid residue having a negative charge; x and y are integers having an independent value of 1, 2, 3 or 4; and n is an integer having the value 1-5.

3. The method of paragraph 2, wherein the self-assembling peptides comprise a sequence of amino acid residues corresponding to formula III or formula IV.

4. The method of paragraph 3, wherein Xaa is alanine, Xaa⁺ means arginine or lysine, and Xaa is aspartic acid or glutamic acid.

5. The method of paragraph 2, wherein the amino acid residues include naturally occurring amino acid residues.

6. The method of paragraph 2, wherein the amino acid residues are naturally occurring amino acid residues.

7. The method of paragraph 1, wherein the self-assembling materials are selected from the group consisting of peptidomimetics, nucleotide, bidentate and triblock copolymers, N-alkylacrylamides and dendrimers.

8. The method of paragraph 7, wherein the self-assembled material is a peptidomimetic.

9. The method of paragraph 8, wherein the peptidomimetics are selected from the group consisting of α-peptides, β-peptides, γ-peptides, δ-peptides and oligomers having primary chains that can assume a helix or card conformation.

10. The method of paragraph 4, wherein oligomers having primary chains that can take on the helix or card conformation are selected from the group consisting of compounds having primary chains utilizing bipyridine segments, compounds having main chains utilizing solvophobe interactions, compounds having primary chains utilizing chain interactions side, compounds having primary chains utilizing hydrogen bonding interactions, and compounds having primary chains using metal coordination bonds.

11. The method of paragraph 7, wherein the self-assembled material is a nucleotide -imultimate.

12. The method of paragraph 11, wherein the nucleotide is selected from the group consisting of isomeric oligonucleotides, modified carbohydrates, oligonucleotides with modified nucleotide linkages, and nucleotides with alternative nucleobases.

13. A method for forming a barrier to the movement of fluids, cells, tissue, biomolecules or gas, comprising feeding to a self-depositing material requiring this site, the self-assembling material selected from the group consisting of peptidomimetics, nucleotide exantics, di- and triblock copolymers, alkylacrylamides and dendrimers.

14. The method of paragraph 13, wherein the self-assembled material is a peptidomimetic.

15. The method of paragraph 14, wherein the peptidomimetic is selected from the group consisting of α-peptides, β-peptides, γ-peptides, δ-peptides and oligomers having primary chains that can assume a helix or card conformation.

16. The method of paragraph 13, wherein the oligomers having main chains that can take on the helix or card conformation are selected from the group consisting of compounds having primary chains using bipyridine segments, compounds having main chains utilizing solvofobes, compounds having primary chains utilizing the interactions side chains, compounds having primary chains utilizing hydrogen bonding interactions, and compounds having primary chains utilizing metal coordination bonds.

17. The method of paragraph 13, wherein the self-assembled material is a nucleotide -imultimate.

18. The method of paragraph 17, wherein the nucleotide is selected from the group consisting of isomeric oligonucleotides, modified carbohydrates, oligonucleotides with modified nucleotide linkages, and nucleotides with alternative nucleobases.

19. A method of regenerating or repairing tissue or tissue-forming cells comprising administering to tissue or cells self-assembling materials, wherein the self-assembling materials are selected from the group consisting of peptidomimetics, nucleotide, bidentate and triblock copolymers, N-alkylacrylamides and dendrimers, alone or in combination. combined with a pharmaceutically acceptable carrier.

20. The method of paragraph 19, wherein the self-assembled material is a peptidomimetic.

21. The method of paragraph 20, wherein the peptidomimetic is selected from the group consisting of α-peptides, β-peptides, γ-peptides, δ-peptides and oligomers having primary chains that can assume a helix or card conformation.

22. The method of paragraph 19, wherein the oligomers having main chains that can assume a helix or card conformation are selected from the group consisting of compounds having primary chains utilizing bipyridine segments, compounds having primary chains utilizing solvophobe interactions, compounds having primary chains utilizing chain interactions on side chains, compounds having primary chains utilizing hydrogen bonding interactions and compounds having primary chains utilizing metal coordination bonds.

23. A method according to paragraph 19, wherein the self-assembling material is a nucleotide-stimulant.

24. The method of paragraph 23, wherein the nucleotide is selected from the group consisting of isomeric oligonucleotides, modified carbohydrates, oligonucleotides with modified nucleotide linkages, and nucleotide nucleotides with alternative nucleobases.

25. The method of paragraphs 1, 13 or 19 further comprising providing one or more therapeutic, prophylactic, diagnostic or cell therapeutic agents.

26. The method of paragraphs 1, 13 or 19 further comprising providing with the self-assembled materials a pharmaceutically acceptable carrier or template, or adjunct material, for administration to the body or the body.

27. The method of paragraph 26, wherein the materials are administered in the form of a dry powder, wafer, disk, tablet, capsule, liquid, gel, cream, foam, ointment, emulsion, suspension, solution, as a coating on a medical device or implant, or in the form of incorporated into microparticles, polymer matrices, hydrogels, textiles, stitches or sponges.

28. The method of paragraph 19, comprising applying to the defect of the cardiovascular or neurological system, wound or area of ​​ischemic necrosis to allow or favor repair of a defect, wound or area of ​​ischemic necrosis.

29. The method of paragraph 19 comprising applying to the bone area with a defect or a wound to enable or promote repair of a defect or wound.

30. The method of paragraph 19 including applying to visceral organs with a defect or a wound to enable or promote repair of a defect or injury.