Combination therapy for the treatment of ocular neovascular disorders

For treating combined treatment of eye novel vascular diseases

[0001] The use is a part of use of the August 26, 2004, and number is 200480031700. 1, and application of invention name and invention same patent of invention application thereof.

[0002] Associated method

[0003] The simulating a random number of the application requirement on August 27, 2003 submitted 60/498,407, attorney file of EYE-013P, a simulating a random number of March, 2004 Mm Ri submitted 60/556,837, the attorney file of mainly EYE-013P2, wherein the literature is punished the article to periphery of the Israeli foot text form.

Invention field

[0004] The invention relates to a pressure and medicine field. A plurality of said that the invention relates to a is able to platelet-derived inhibiting growth factor (PDGF) and vascular endothelial growth factor (VEGF) reagent combined treating eye novel vascular diseases.

[0005] invention background

[0006] Blood vessel for; the dart and newborn vascularization, comprising a extant vascularization novel (sprout), and they invades the external tissue. A related process, blood vessel is (vasculogenesis) comprises already to exist of the tissue the endothelial battery and the differentiation of blood vessel battery, and they ring afterward in turn to form a blood vessel.

[0007] The vascular occurred widely can during the grower, and can also during wound healing of the building body, to in the blood to flow to the tissue of the injury or the injury. Natural, the blood vessel occurred is formed associated with cancer and tumour. The fact, wherein tumour tissue the number of the vessel is breast cancer (Weidner and so on, (1992) J. Natl. Cancer Inst. 84: 1875-1887), Prostate cancer (Weidner and so on, (1993) Am. J. Pathol. 143: 401-409), And tumour (l2 and others, (1994) Lancet 344: 82-86) And melanoma O7Oss and so on, (1996) Res Cancer. =2900-2903 56) Powerful negative prognosis target. Recently discovered for blood vessel occurred with the other disease is composed of multiple medicine field of related, comprising rheumatism extract, Dermatologica, heart disease studying and ophthalmology. Specifically speaking, does not hope or the tissue for specific blood vessel occurred with a special disease is related, card further comprises a rheumatoid arthritis atherosclerosis, and psoriasis for example, Fan, (19%) Trends Pharmacol. Sci. 16: , 57With Folkman (1995) And Med. 1: 27). Moreover, the blood vessel permeability the is considered with an effect (Cullinan-Bove the normal and for physiological process and so on, (1993) Endocrinol. 133: , 829Senger and so on, (1993) Cancer and Metastasis Reviews 12:303). Although for processing photo occurred with a growth blood vessel of the spherical crown disease with a tumour blood vessel occurred to and multiple treatment, wherein each one possibly with the special, the periphery the straight of cell with.

[0008] A diseases and steering for blood vessel with. For example, diabetic retinopathy, a of the third large end of the adult of a sight (accounts for blind population of the simulating almost 7%), a widespread for is related. The non- hyperphasia retinopathy the pericyte's the selectivity with a retina, and their forfeits is covered on the telescopic of related blood tubular, and arced adding of blood flow. The blood tubular of stretching, endothelial battery and; and form, comprising a fine the moving block, and adjacent blood tubular is blocked, so the formed on the external retina area of not poured into. Chinese, leg blood vessel can of the adjacent fine motion block region, and a card with an motion block early diabetic retinopathy and non- irrigation the clinical image of retina area. An motion leakage block, and blood tubular possibly, an of the exudate and hemorrhage. At which with the background diabetic end retinopathy's end, wherein the conditions to developing for multiple years the time, develops the hyperphasia diabetic retinopathy, and left 5% blind case. When the multiple regions of annular retina to a usage blood tubular and cannot injecting, a water of the other retina plate and base is a new blood vessel, comprising a hyperphasia diabetic retinopathy. Are new blood connector grow the vitreous body, and easy to release, thus front make the retina, a release. The period in hyperphasia diabetic retinopathy, the majority of block vitreous body hemorrhage possibly ring vitreous chamber. Moreover, the new blood vessel with nano edible-medicinal fibre structure, comprising possibly the traction removing of retina.

[0009] Diabetic retinopathy is main and diabetic duration is related; Therefore, along with is longer, diabetes retinopathy's prevalence for five-high age and bottle or the reel. The laser therapeutic is used for non- hyperphasia and hyperphasia diabetic retinopathy is. The yellow area of the infection laser therapeutic leakage of essential motion block sleeve is made of clinical remarkable yellow and dropsy forfeit of patient vision to 50% and. The hyperphasia diabetic retinopathy, the whole retina light coagulation sleeve are multiple external a small (to avoid yellow area of) on the whole retina; The pair of treatment of skin blind 60% of. A for early treatment of yellow and dropsy and hyperphasia diabetic retinopathy for preventing patient 95% to a a sight of five years, and period in treating capable of preventing 50% to patient and a sight. Therefore, early diagnosis and treatment of substances.

[0010] The infant vascularization the eye disease of the bumps relevant yellow part denaturates (AMD), the is a affects left 1/10 American's more bumps 65 of diseases. The AMD the yellow part, namely a a switch for of changing of retina central region, the pair of changing with large the visual acuity; the affects the central vision. AMD claims a dart the retinal epidermis pigment single-layer battery, a screwed on the temperature of the retina. Are cells nourish, and support and retina part of one person, namely, comprising television to the pigment the visual panel. The retinal pigment intraepithelial is located at the glass layer, which is a basilar composite film; the AMD patient, comprising a aggregation, and hardens. New blood vessel possibly from the choroid of glass layer, comprising a rich blood vessel bed. Are blood connector is possibly divulge the liquid or release of the retinal pigment intraepithelial, and is an possibly between the retinal pigment epidermises and sensor retinas. The afterward fibre scar layer of prodrugs of the visual nutrient, and arced are battery deaths, thus such forfeit of the visual acuity. The type's people yellow relevant pathological part of the dart and a thereof in manner; the is a blood vessel and retina for minimizing the dropsy or blood. The humidity-discharging type of accounts of the bumps relevant yellow of pathological the case 10%, wherein a sleeve of the senior 90% citizen case, can the yellow part denaturates, and is legal blind. A dry on the age type relevant yellow of pathological the relates to a decomposition of retinal pigment intraepithelial, and forfeit of the visual panel. Dry type's pathological changing the skin vision, and usually with 20/50-20/100 layer.

[0011] AMD with deformation of the vision, compensating the field of increasing or the small or the straight line in deformation, the or without Zhongxinduan. The humidity-discharging type's AMD, possibly notes to feel small is separated from the retina of the yellow point area; further, a retina the blood vessel film chinese diagnosis to be the fluorescein motion operation. Wherein the drying type, a glass wart possibly disturbed the pigmentation mode of yellow area of. The glass wart of the retinal epidermis pigment basilar film goitre, which can with a battery prominently, so it is connected with the battery is a telescopic; They are the bumps relevant yellow of pathological change's hazard factor end and is still cleaning. The current for treating dry and method of age of relevant yellow of pathological the. The laser therapeutic is used for the humidity-discharging and age relevant yellow of pathological change, and eliminated for newborn blood vessel of the first, and prevented the left 50% patients' further vision and output when 18 months. Natural, wherein to 60, months cover and 20% patient still with a remarkable effect.

[0012] Already appraises multiple types of molecule mediators of the vessel is, comprising for and acidic fibroblast growth factor (aFGF, bFGF), transforming growth factor α and β (TGFa, TGF β), platelet-derived growth factor (PDGF) and angiogenin and blood platelet derivation battery endothelial growth factor (PD-ECGF) and Interleukin one side (IL-8) and vascular endothelial growth factor (VEGF). Participates the other stimulants of the blood collecting pipe angiogenin (angiopoietirO-LDel-l and graafian follicle analogue-polyethylene (follistatin) and granulocyte group of stimulating (G-CSF) and liver battery growth factor (HGF), thin protein, midkine and placenta growth factor and multi-effect protein (PTN), progranulin and protein and Tumour Necrosis Factor and a (TNF-a). Moreover, multiple types of said modulators for the control of the vessel is can also through is produced vascular with a body, lead (fibrin dissolves connected zymogen) comprises angioarrestin and vascular analogue-polyethylene, the antiangiogenesis an zymoplasm III and cartilage and inhibitor of derivation ((3 )I), (3 )59 complement connected and bast inhibin (endostatin) (collagen XVIII bonding), fibronectin connected and gro- β, heparinase joint and hexasaccharide connected and human choriogonadotropin (hCG) and interferon α/β/Υ and interferon induction protein (ΙΡ -10), Interleukin -12, kringle (5 dissolves fibrin zymogen connected), metal protease inhibitors (TIMPs) and 2_A oxygen female keep the diol and a placenta ribonuclease inhibitor and fibrin to the aerosol zymogen of inhibitor and platelet factor and (4 PF4), prolactin 16kD connected and protein and related protein (PRP), a of substance and four hydrogen cortisols of S and blood platelet reaction Protein and i (tsp-i) and vessel analogue-polyethylene (vasculostatin) and blood vessel inhibiting factor (vasostatin) calcium (reticulin connected).

[0013] The modulator display crucial role of the upper for modulators, the VEGF and achievement following the tumour growth and blood vessel with (summary card further Brown and so on, (1996) of Control For (Goldberg and Rosen, Eds). Birkhauser, Basel, and Thomas (1996) J. Biol. Chem. 271: 603-606). The evening divination, the studied recently already signal conductive molecule PDGF family the function of PDGF-B it can, comprising and multiple, the moulding of perivascular battery with the normal functional and educational a role, sometimes wherein the perivascular battery is dart and wall battery, for example, blood vessel smooth rib and renal glomerulus film battery and pericyte.

[0014] Although to form a blood vessel is soaked with or the understanding of the vascularization with the grower, wound healing and tumour, it still needs to determine; the blood vessel occurred with the eye blood vessel occurred whether with extending. Very display, following freely, for example, a vascular on the cross support vascularization occurred to the biological is possibly convenient, and adaptation biological, and for example the AMD pathological eye newborn vascularization and known, and is usually to adjust and a sight (the related summary card further Campochiaro (2000). J Battery. Physiol. 184: 301-10). Therefore, although to the following vascularization the understanding of molecule event with already made the further, it still needs to use are performed to developing for treating other method of novel vascular diseases, wherein disease comprising a novel vascular diseases and such as with a disease according choroid of the vascularization AMD and diabetic retinopathy soaked with.

[0015] invention outline

[0016] Already surprisedly discovered is sectional area of vegf and sectional area of pdgf the combination of agent, and a matched with the treatment and when the treating eye novel vascular diseases.

[0017] Therefore, the invention said for treating is diagnosed to cover or with the emergency to developing a novel vascular diseases the method of patient. The method comprises the patient to employ To And vegf agent and Resisted and pdgf agent, a main or adjuvant for treating.

[0018] At the same time, the invention provides for restraining with the patient and method for of new vascular diseases steps, which is simultaneously or separated belt is about 90 to days employ the PDGF antagonists and VEGF antagonists to patient and realizes, the buffer for suppressing with a novel vascular diseases foaming.

[0019] On the other handle, the invention provides for treating with the method for of patient steps, wherein the patient is diagnosed to cover or with the emergency to developing is a novel vascular diseases, comprising a or separated belt is 90 to days employ the PDGF antagonists and VEGF antagonists to patient and realizes, to the buffer are capable of sufficiently patient.

[0020] The specific by plans of said aspects, methods of the invention comprising a convex part is about 10 days to employ the PDGF antagonists and VEGF antagonists of. The other preferred embodiment of the invention method, wherein the PDGF antagonists and VEGF antagonists is fixed into the is 5 to days of employ. The other preferred embodiment of the invention method, wherein the PDGF antagonists and VEGF antagonists is fixed into the is about END to method of employ. The invention method's specific preferred embodiment, wherein the PDGF antagonists and VEGF antagonists to be employ.

[0021] The other preferred embodiment, method of this invention comprising administering an PDGF antagonist, which is a PDGF-B antagonist. The other preferred embodiment, method of this invention comprising administering an VEGF antagonist, which is a VEGF-A antagonist.

[0022] The multiple by plans, method of this invention comprising administering an PDGF antagonist, which is a nucleic acid molecules, the surface body and antisense RNA molecule, ribozyme, the RNAi molecule, protein, peptide, the ring peptide, antibody and antibody connected and connected, sugar, polymers or the small organic compound. The other preferred embodiment, method of this invention comprising administering an VEGF antagonist, which is a nucleic acid molecules, the surface body and antisense RNA molecule, ribozyme, the RNAi molecule, protein, peptide, the ring peptide, antibody and antibody connected and connected, sugar, polymers or the small organic compound.

[0023] The specific preferred embodiment, method of this invention comprising administering an VEGF antagonist, which is a telescopic body, to EYE001 surface body. The other preferred embodiment, method of this invention comprising administering an VEGF antagonist, which is a antibody or a connecting connected.

[0024] The specific preferred embodiment, method of this invention comprising administering an PDGF antagonist, which is a telescopic body, antibody or a connecting connected. The other specific embodiment, method of this invention comprising administering an PDGF antagonist, which is a antisense oligonucleotide.

[0025] The invention the; the handle and other preferred embodiment, wherein the PDGF antagonists and/or the VEGF antagonists of the front medicine.

[0026] A preferred embodiment, methods of the invention provides for restraining or treating the method of eye novel vascular diseases. The multiple by plans, suitable for through the novel eye vascular diseases according a method for the invention treating or pressed comprising a ischemic retinopathy and iris newborn vascularization, the intraocular newborn vascularization and bumps relevant yellow part denaturation and cornea newborn vascularization and retina newborn vascularization and choroid newborn vascularization and diabetic retina ischemic or the hyperphasia diabetic retinopathy. The other preferred embodiment, methods of the invention provides or diagnosed to cover or with the emergency to developing for the patient restraining or treating with the steps of the disease patient psoriasis or a method of rheumatoid arthritis.

[0027] The invention claims a further comprises a PDGF antagonists and VEGF antagonist, and capable and supporting pharmaceutical composition. On the surface, PDGF and amount of use of VEGF antagonists foaming pressed with a novel vascular diseases.

[0028] The; a handle of a preferred embodiment, wherein pharmaceutical composition comprising a PDGF antagonist, which is a PDGF-B antagonist. The other preferred embodiment, wherein pharmaceutical composition comprising a VEGF antagonist, which is a VEGF-A antagonist.

[0029] The multiple by plans, the pharmaceutical composition of the invention comprises a PDGF antagonist, which is a nucleic acid molecules, the surface body and antisense RNA molecule, ribozyme, the RNAi molecule, protein, peptide, the ring peptide, antibody and antibody connected and connected, sugar, polymers or the small organic compound. The other preferred embodiment, the pharmaceutical composition of the invention comprises a VEGF antagonist, which is a nucleic acid molecules, the surface body and antisense RNA molecule, ribozyme, the RNAi molecule, protein, peptide, the ring peptide, antibody and antibody connected and connected, sugar, polymers or the small organic compound.

[0030] The specific other by plans, the pharmaceutical composition of the invention comprises a VEGF antagonist, which is a telescopic body, to EYE001 surface body. A preferred embodiment, the pharmaceutical composition of the invention comprises a VEGF antagonist, which is a antibody or a connecting connected.

[0031] The specific preferred embodiment, the pharmaceutical composition of the invention comprises a PDGF antagonist, which is a antibody or a connecting connected. The other specific embodiment, the pharmaceutical composition of the invention comprises a PDGF antagonist, which is a antisense oligonucleotide.

[0032] The pharmaceutical composition of the invention comprises a can be for supporting, comprising a small ball or hydrogel preparation.

[0033] The other preferred embodiment, wherein the PDGF antagonists and/or the VEGF antagonists of the front medicine.

[0034] The other preferred embodiment, the pharmaceutical composition of the invention provides for restraining or treating the method of eye novel vascular diseases. The multiple by plans, suitable for through the novel eye vascular diseases according a pharmaceutical composition of the invention treating or pressed comprising a ischemic retinopathy and iris newborn vascularization, the intraocular newborn vascularization and bumps relevant yellow part denaturation and cornea newborn vascularization and retina newborn vascularization and choroid newborn vascularization and diabetic retina ischemic or the hyperphasia diabetic retinopathy. The other by plans, the pharmaceutical composition of the invention is provided with a patient of steps, or is diagnosed to cover or with the emergency to developing or used for treating psoriasis for the disease patient on or the method of rheumatoid arthritis.

[0035] The invention claims a further comprises a PDGF antagonists and medicine bag of VEGF antagonists (pharmaceutical overlapping). A preferred embodiment of the surface; the medicine bag comprising a PDGF antagonist, which is a PDGF-B antagonist. The other preferred embodiment of the surface; the medicine bag comprising a VEGF antagonist, which is a VEGF-A antagonist.

[0036] The other preferred embodiment, wherein the medicine bag PDGF antagonists and VEGF antagonists is a, and preparation of the independent dosage form. The other preferred embodiment, wherein the medicine bag PDGF antagonists and VEGF antagonists preparation of.

[0037] A specific by plans, a medicine bag of the invention comprises a VEGF antagonist, which is a telescopic body, to EYE001 surface body. The other by plans, a medicine bag of the invention comprises a VEGF antagonist, which is a antibody or a connecting connected.

[0038] The multiple by plans, a medicine bag of the invention comprises a PDGF antagonist, which is a antibody or a connecting connected. The specific other by plans, a medicine bag of the invention comprises a PDGF antagonist, which is a antisense oligonucleotide. The other preferred embodiment of the surface; the PDGF antagonists and/or the VEGF antagonists of the front medicine.

[0039] Joint between summary

[0040] Chart I (A) of the person PDGF-B nucleic acid sequence (GenBank which X02811) SEQ (ID NO: 1) Schematic drawing.

[0041] Special 1 (B) of the person PDGF-B amino acid sequence (GenBank which CAA^579) SEQ (ID NO: 2) Schematic drawing.

[0042] Chart I (C) is a of PDGF-A nucleic acid sequence (GenBank which X06374) SEQ (ID NO: 11) Schematic drawing.

[0043] Chart I (D) of the person PDGF-A polypeptides sequence (GenBank which CAA29677) SEQ (ID NO: 12) Schematic drawing.

[0044] Special 2 (A) is a of VEGF nucleic acid sequence (GenBank the number: NM_003376) SEQ (ID NO: 3) Schematic drawing.

[0045] Special 2 (B) is a of VEGF polypeptides amino acid sequence (GenBank which NP_003367) SEQ (ID NO: 4) Schematic drawing.

[0046] Special 3 (A) is of PDGFR-B nucleic acid sequence (GenBank which NM_002609) SEQ (ID NO: 5) Schematic drawing.

[0047] Special 3 (B) is of PDGFR-B polypeptides sequence (GenBank which NP_002600) SEQ (ID NO: 6) Schematic drawing.

[0048] Dumbbell 3 (C) is a PDGFR-A nucleic acid sequence (GenBank which NM_006206) SEQ (ID NO: 13) Schematic drawing.

[0049] Special 3 (D) of the PDGFR-A polypeptides sequence (GenBank which NP_006197) SEQ (ID NO: 14) Schematic drawing.

[0050] Special 4 (A) of the person VEGFR-I (Flt-I) nucleic acid sequence (GenBank which AF063657) SEQ (ID NO: 7) Schematic drawing.

[0051]Special 4 (B) of the person VEGFR-I (Flt-I) polypeptide sequence (GenBank the number) SEQ (ID NO: 8) Schematic drawing.

[0052] Special 4 (C) is a of VEGFR-2 (KDR/Flk-1) nucleic acid sequence (GenBank which AFO! 35121) SEQ (ID NO:

9)schematic drawing.

[0053] Special 4 (D) of a person VEGFR-2 (KDR/Flk-1) polypeptide sequence (GenBank which AAB88005) SEQ (ID NO:

10)schematic drawing.

[0054] Special 5 is compare the comparison to (cont), Gleevec processing (sectional area of pdgf agent) and Macugen™ processing (i.e. pegaptanib processing, Resists and vegf agent the cornea newborn vascularization measuring result, and (sectional area of pdgf/with Macugen™ and Gleevec combined process and sectional area of vegf combined treating) graphical representation.

[0055] Special 6 (8) can be in comparison (PEG- processing) mouse cornea cornea newborn vascularization the photo graphical representation is of fluorescence microscope image.

[0056] Special 6 (B) can the Gleevec- processing mouse cornea cornea newborn vascularization the photo graphical representation of a of fluorescence microscope image.

[0057] Special 6 (C) can the Macugen™- processing mouse cornea cornea newborn vascularization the photo graphical representation of a of fluorescence microscope image.

[0058] Special 6 (D) can mouse cornea cornea newborn vascularization the photo graphical representation on the fluorescence microscope image, and is Macugen™ and Gleevec two treatment.

[0059] Special 7 (A) is fluorescence microscope image photo graphical representation, expressed of the normal cornea vascular system is not the bearing employ (APB5 the PDGFR antibody, Resists and pdgf agent) force.

[0060] Special 7 (B) is fluorescence microscope image photo graphical representation, expressed of the normal cornea vascular system are not employed the straight of Gleevec.

[0061] Special 7 (C) is a fluorescent microscope image photo graphical representation, expressed of the normal cornea vascular system are not be employed Macugen™ (Mac) and force of Gleevec.

[0062] Special 7 (D) is fluorescence microscope image photo graphical representation, expressed of the normal cornea vascular system are not employed the straight of PEG.

[0063] Special 8 is laser sensing the graphical representation of choroid newborn vascularization measuring result, comprising processed (cont) preferably the comparison, Gleevec processing (sectional area of pdgf agent) and Macugen™ processing (i.e. pegaptanib processing, Resists and vegf agent) and a Macugen™ and Gleevec combination processing and (sectional area of pdgf/sectional area of vegf combined treating).

[0064] Special 9 is laser sensing the graphical representation of choroid newborn vascularization measuring result, preferably comparison - processing (cont) and APB5- processing (sectional area of the pgfr antibody, and educational Is Resistance of pdgf a role of agent) and Macugen processing (i.e. pegaptanib processing, Resists and vegf for body) and (Mac+APB5) is processed with Macugen and APB5 and.

[0065] Special 10 is a retina growth model and graphical representation, comprising processed (Cont) and ARC-127 processing (To And pdgf preferably the comparison agent) and Macugen processing (i.e. pegaptanib processing, Resists and vegf agent) and (sectional area of pdgf/with Macugen and ARC-127 combination processing sectional area of vegf combined treating) and.

[0066] Special 11 is a cornea newborn vascularization measuring result graphical representation, comprising processed (cont) and ARC-127 processing (To And pdgf preferably the comparison agent) and Macugen processing (i.e. pegaptanib processing, Resists and vegf agent) and (sectional area of pdgf/with Macugen and ARC-127 combination processing sectional area of vegf combined treating) and.

[0067] Special 12 (A) can comparing the mouse cornea cornea newborn vascularization the photo graphical representation of fluorescence microscope image.

[0068] Special 12( 3) can the ARC-127- processing mouse cornea cornea newborn vascularization the photo graphical representation of fluorescence microscope image.

[0069] Special 12 (C) can the Macugen- processing mouse cornea cornea newborn vascularization the photo graphical representation of fluorescence microscope image.

[0070] Special 12 (3) can mouse cornea cornea newborn vascularization the photo graphical according representation of fluorescence microscope image, and is Macugen and ARC-127 process.

[0071] Special 13 is a cornea newborn vascularization measuring result graphical representation, comprising processed (cont) and APB-5 processing (To And pdgf preferably the comparison agent) and Macugen processing (i.e. pegaptanib processing, Resists and vegf agent) and (sectional area of pdgf/with Macugen and APB-5 combination processing sectional area of vegf combined treating) and.

[0072] Special 14 is a cornea newborn vascularization measuring result graphical representation, comprising processed (cont) and APB-5 processing (To And pdgf preferably the comparison agent) and Macugen- processing (i.e. pegaptanib processing, Resists and vegf agent) and (sectional area of pdgf/with Macugen and APB_5 combination processing sectional area of vegf combined treating) and.

[0073] Invention and

[0074] Is punished the article to periphery of said literature, patent and patented claim of each substituting mentioning.

[0075] ^JL

[0076] The article, the following terminology and phrase with the semantic with. Technique thereof and scientific terms of a plurality of can, the article for is sealed with the invention the technical field claims a personnel generally the same meaning of understanding.

[0077] A antagonists of said of the part or inhibiting reagent for high molecular the activeness or production of. Specifically speaking, wherein terminology a antagonists, wherein the paper selectively use, preparation of according to PDGF, PDGFR, VEGF or the VEGFR gene expression layer, the mRNA layer, a protein or protein active reagent. The showing form of antagonists comprising, for example, protein, polypeptides and peptide (for example a peptide), antibody or antibody connected, peptide use, nucleic acid molecules, antisense molecule, ribozyme, suitable body, RNAi molecule and small molecular organic. The antagonists for inhibiting VEGF/VEGFR and PDGF/PDGFR ligand/receptor improves the non- definition mechanism of demonstration's comprising the pressing ligand body and stability (e.g., a target location groove ligand gene/nucleic acid antisense, ribozyme or the composition RNAi), ligand's is connected with the homologous receptor (e.g. use, resists is suitable ligand body, antibody or solubility in homologous receptor), to suppress the receptor body and stability (e.g., a target location groove receptor ligand gene/nucleic acid antisense, ribozyme or the composition RNAi), receptor is connected with the homologous receptor (e.g., a receptor antibody) and is the receptor of the activation by wherein homologous ligand(E.g., a receptor tyrosine kinase inhibitors). Moreover, wherein the antagonists to be directly or indirect inhibition target molecules.

[0078] The article, terminology of antibodies in intends to comprising a of antibodies, for example, a homogeneous antibody (IgG, IgA, IgM, IgE), and a are respectively connected parallel, and can also with the vertebrates (e.g., mammal) protein, carbohydrate and other specificities with the reaction. The antibody capable of folium artemisiae technology connected, and screen are fragments of the upper cover antibody same and use. Therefore, the terminology comprising antibody molecule's the part or connected to preparation of proteinase cutting, which can with multiple proteins selectively the reaction. The non definition of enzymatic protein or connected to a Fab, the F ab' (2), Fab', Fv and a stranded antibody (scFv), comprising a peptide joint and V-SHAPED (L) and/or THE [PART OF field. The scFv' s to be covalent or the non- covalent connected, and aim of formed with two or more than two binding sites antibodies. The invention includes the other purifying preparation and antibodies to polyclonal of antibodies, monoclonal antibody or antibodies.

[0079] Here, terminology is suitable body, wherein terminology with a nucleic acid ligand an alternating-current use, comprising the outer nucleic acid, wherein the nucleic acid adjust the special three three-dimensional conformation ability by, comprising a fixed light of the target-type molecule, and antagonistic of the bracket (i.e., limited). Target of the invention is PDGF or VEGF (or a of every homologous receptor PDGFR or VEGFR), accordingly, comprising a terminology the PDGF surface body or a nucleic acid ligand or the VEGF surface body or a nucleic acid ligand (or PDGFR surface body or nucleic acid ligand or VEGFR surface body or nucleic acid ligand). The telescopic body and a target through the following manner to carry and: And the target, the taking the target, to decorate/of the function of active groove and target-type of target or target with the reaction, and committing suicide the inhibitor and target-type covalent which is attached, prompting responses between the other targets and molecules. For a body comprises multiple ribonucleotide unit and contains units or are two type invention residues mixtures. For body is a comprises one or more with decorative the base group, sugar or phosphate the principal chain unit, common to further claims the paper.

[0080] An antibody antagonists of expressed the antibody molecule of the article defines, comprising a block or display weaken target PDGF or VEGF a or multiple types of activeness. For example, the VEGF inhibiting antibody and inhibiting or weaken ability for VEGF stimulation blood vessel with.

[0081] Such as each base group of two sequences matched, which can form watson and Krick (Watson Crick) is pair, comprising a polynucleotide sequence and is one of a polynucleotide sequence of the co-grinding. Terminology a supplementary chain a here of terminology of each complement an alternating-current use. Nucleic acid an are complement can be coding strand's complement or non-coding including complement each other.

[0082] Phrase a conservative residue shape or a conservative amino acid substituted a expressed based on a common treatment amino acid grouping. The common treatment functional arranged between 2-4 independent amino acids is detachably homologous biological of proteins the normalized frequency of the amino acid the. According to the analysis, for determining amino acid group, amino acid in the group is of each other, or therefore, the person to the straight of total protein structure of each other trench (Schulz, G.E.R.H. Schirmer, Principles of Protein Structure, Springer-Verlag). The freestanding grain groove Shijiao by such device of amino acid group of comprises:

[0083] (1) With the electric charge's group, composed of Glu and Asp, Lys, and Arg, Which

[0084] and (II) with positive charge's group, composed of Lys, and Arg, Which

[0085] (iii) with a negative charge group, and Glu and Asp composition,

[0086] the IV) aromatics group, composed of Phe, Tyr and Trp,

[0087] (ν) nitrogen ring group, composed of Which and Trp,

[0088] the vi) are connected aliphatic non-polar group, composed of Val, Leu and Ile,

[0089] the vii of) are set, comprising a Body and Cys,

[0090] (viii) small residue-filter assembly, composed of kr, Thr, Asp, Asn, Gly, Ala, Glu, Gln and Both,

[0091] the ix) aliphatic one group, composed of Val, Leu, upper, and Is Cys, a

[0092] (χ) small hydroxyl group, composed of Ser and Thr.

[0093] The group of removing is enumerates, wherein a of amino acid residue capable of wherein of group, and a tension device is formed by the independent amino acid is provided with a single the abbreviations of letters and three letters of the field of commonly amino acid

[0094] The article, terminology the shape of the fact expressed the molecules detectable the micro-processing or the connecting body (e.g.. the shape of), to protein of protein and protein and nucleic acid and nucleic acid and nucleic acid and protein and small molecular or nucleic acid the shape between the small molecules.

[0095] Terminology of protein shape of the preparation of the other with the field interactions protein, unions and/or is arranged on the proteins, for example, PDGF or VEGF protein, or parallel corresponding homologous receptors.

[0096] The article, a nucleic acid, multiple DNA or VACCINE the corresponding terminology a layer, expressed with existing on the other DNAs or a molecule of the RNAs are respectively macro-molecule natural source. Production, wherein the paper, a expressed of protein molecule with the origin of comprising FGF21 mutant polypeptides corresponding terminology a layer of the proteins for separating. The paper, terminology separating further expressed of wherein when the recombinant DNA production technology basically and comprising a battery material; the virus material or culture medium nucleic acid or a peptide, or is basically and other gametocide precursors or chemical reagent by the chemical body.

[0097] A nucleic acid for separating the expressed of a nucleic acid connected, comprising not exist and small connected, and is not discovered of the endoscope. Terminology a separating plate is also used for lighting polypeptide here, comprising separated from the battery proteins, and expressed comprising a purifying and to comprising two.

[0098] The article, terminology of a mark and capable of detecting mark to indicate of the detection molecule, comprising, and not limited, same isotope, fluorophore and chemiluminescence part, enzyme and enzyme substrate, enzyme cofactor and enzyme inhibitor, dye, metal ion and ligand (e.g., idrabiotaparinux or haptin). Terminology a fluorescer preparation of capable of displaying can be detection of fluorescent material or wherein the. A comprising a fluorescein, rhodamine and Each support acid, umbellulone and Texas is red of the foundation of the cigarette of the invention is, luminol, NADPH and α and β - half part glycosidase and horseradish peroxidase.

[0099] A gene's type layer of a battery expressed is composed of the battery mRNA and coding gene, front and mRNA, for newborn duplication, the duplication processing middle, mature mRNA and a layer of degradation product, and is of the gene control a layer of protein.

[0100] The article, terminology a nucleic acid expressed of the polynucleotides, which DNA (DNA), and which is proper, further - RNA expressed (RNA). The terminology further can is understood comprising a nucleotide interleukin-11 VACCINE preparation or the DNA interleukin-11 and equating article, and is arranged above to the preferred embodiment, the single stranded (with righteousness or antisense) and a including polynucleotides, ESTs, chromosome, cDNAs, mRNAs and rRNAs of the dart and a nucleic acid the representative example of molecules.

[0101] Terminology a oligonucleotide a expressed of nucleotide or the nucleoside monomer's widowed a body or the polymer, wherein the principal chain) key composed of a base group carbohydrate and a candy with small. The terminology farther comprising a decorative or with substituted for widowed a body, comprising a single or wherein part of the nonnatural, they are provided and a role. Of substituted for conformity of widowed a body is provided with a factors, comprising a reinforcing the battery ingestion, or reinforcement nuclease resistance, and according to the field of male knowledge the method. The angle oligonucleotide or cover is wherein a part of a (widowed a body.

[0102] Terminology a percentage identity in sequence expressed identities between two types of amino acid sequences or the two types of polynucleotide sequences. The identity of the position of each sequence of determining through the comparison with they can be is compare for comparison. When the same position in sequence hypotensive when the same base group or the amino acid, wherein the molecule of the position of the; The same position of the same or similar amino acid residue (e.g.. when space and/or electronic and production water), wherein the molecule is dart and positions of the homology (production). Homology, similarity or identical the percentage expressed by comparison sequence of the total of same or similar amino acid the function of. Capable of each or preferably to algorithm and/or the it, comprising a Hidden Markov Model (HMM), FASTA and BLAST. HNiM and FASTA harvester grain opening BLAST of obtain from the following: mechanismthe Metal Centre is Biotechnology Information and Metal Library of Medicine and Metal of The Health, Bethesda, Md. harvester by the opening European Bioinformatic Claims EBI. A preferred embodiment, the percentage identities of two types of sequences is connected with the GCG for determining to the wherein the weight is 1, for example, wherein on the weighting to each amino acid gap, is provided to which is single amino acids or the invention mispairings between two sequences. The technical descriptions is in comparison the following literature: Method of Enzymology, vol. 266: Computer Method for Macromolecular And Sequence (1996), ed. Doolittle, Academic Filtering, Inc., a line of Harcourt Support & Co. , San Diego, Acquiring, simulating. Such as or; the hole in sequence with the gap preferably to the it is used for preferably to the sequence. Smith Waterman is permission for type preferably with algorithm of notch (card further (1997) Meth.Mol.Biol to the sequence. 70: 173-187). Similarly, wherein Needleman and 1Wunsch preferably be used to a method for GAP preferably to the sequence. Comprising is HMM multiple technologies and algorithms described of the following literature Sequence, Structure, And Databanks: A Practical Device (2000), ed. Oxford College Filtering, Incorporated and Bioinformatics: Databases and Systems (1999)ed. KluwerAcademic Publishers. The pair of retrieval strategy with a MPSRCH the soft, the soft movable MASPAR computer. MPSRCH for Smith-Watermnan algorithm to shift the sequence on the block parallel machine. The improved method the ability of the micro-processing far matching, and special ability is staggered of the small) and a polynucleotide sequence. Be used to retrieve amino acid sequence of nucleic acid and protein and DNA database. With a database description of being sequence of the following literature: Method of Enzymology, Ed. Doolittle, with. Database comprising Genbank, EMBL and Japan's DNA database (DDBJ).

[0103] An angle of and expressing transfusion DNA, expressed is formed is the transfusion DNA's polynucleotide or a chain oligonucleotide each other provided with a including structure, to the invention each each chain on the invention with the chain with watson and Krick base group of. The terminology of further comprises a pair of nucleoside analog a medicine, to the - inosine, comprising 2) on treating groups nucleosides and equal. The target-type polynucleotides and oligonucleotides or polynucleotides the mispairing between the transfusion DNA expressed of the pair of nucleotide a transfusion DNA and is able to be watson on the Krick unified. The mentioning three chain body, the terminology expressed of three chain body composed of a transfusion DNA and third chain of pair; each invention has Hoogsteen or the connecting Hoogsteen connected with pre-formed a bottom pair on the transfusion DNA.

[0104] Terminology on in VACCINE) and a RNAi shape or a siRNA a expressed by for leading of the target cell to weaken one or more of double-bunch RNAs the gene or gene product with a method of type, wherein RNA and field gene (homology with the field gene, for example, PDGF or VEGF messenger VACCINE) homology.

[0105] Polymorphism raised form of further comprises a single-nucleotide polymorphism body (SNPs); the polynucleotide sequences with a of the base group (e.g., or VEGF variable in a base group of PDGF). Existence of SNPs can be, for example, multiple groups and disease or in disease and target-type of dynamic.

[0106] Exceptionally of the nozzles (a), for example, tumour the biological and expressed by diseases and battery the contents of each ingredient. Battery device comprises a RNA layer, abundance protein or protein active layer.

[0107] Here, terminology of protein shape of terminology with a peptide) and a comprising an alternating-current use. Terminology of protein to a expressed by the invention of recombinant DNA production technology, protein, and is generally the type protein or VACCINE DNA is appropriately the type vector, wherein the type vector is used for variable the main battery, a aim of producing heterologous protein or VACCINE. Moreover, wherein phrase a stemming from an, wherein the left and protein's to ornamental gene, refers to a to a protein of the semantic, comprising with the amino acid sequence of natural protein, or with the provided with amino acid sequence protein of the sudden the production, wherein the sudden the comprising a substituted and flaw of protein with small.

[0108] The article, terminology a gene in expressed of the nucleic acid sequence (device for example, a of target nucleic acid, or wherein antisense duplication and); the already and a nucleic acid sequence to an iontophorectic battery. Is inducting wherein gene animal or battery, a gene capable of the part or completely heterologous, namely, extraneous source, or with inducting wherein gene animal or the endogene gene homology, wherein, they are designed Into Huo with the genomic by such that is, and aim of changing with inserting wherein the genomic (e.g., wherein mounting position of different from natural gene position, or wherein inserting sleeve to knocking removing). A gene further can exist in the form of episome of the pool. A gene and comprises one or more duplication regulation sequence, and one other nucleic acid, such as intron, they possibly of applies the best type of nucleic acid is formed.

[0109] A novel vascular diseases and preparation tightening tube or with a adjustable blood vessel is provided with the disease, carcinogenicity wire or a disease of neoplastic conversion, namely, cancer is an exception. The new vascular diseases comprising psoriasis, rheumatoid arthritis and eye novel vascular diseases, and relevant people yellow part denaturation comprising diabetic retinopathy.

[0110] The article, terminology of the vascularization) and a blood vessel occurred an alternating-current of use. The infant vascularization and blood vessel occurred to indicate for generating new blood vessel to with a battery, tissue or the organ. The control of the vessel is usually with composed of multiple and disease, and reproduction occasions, which diseases associated for injury with tube, and adjustable or the uncontrolled blood vessel occurred related. Disclosed . Moreover the adjustable blood vessel with the socket multiple types of the disease, comprising a finger battery endothelial growth deformity and disease, and supporting the pathology injury of the conditions are provided, comprising and leakage permeability of blood vessel.

[0111] A novel eye vascular diseases shape of the patient eyes changed or with a adjustable blood vessel occurred and disease. Of eye novel vascular diseases comprising optic nerve plate newborn vascularization and iris newborn vascularization and retina newborn vascularization and choroid newborn vascularization and cornea newborn vascularization and vitreous body newborn vascularization, green cataract, pannus, pterygium, yellow and dropsy, diabetic retinopathy and diabetic yellow and dropsy, blood vessel original and retinopathy, retina denaturation, uveitis, retina inflammatory disease and hyperphasia vitreous body retina pathological the.

[0112] Terminology the treatment of subject's novel vascular diseases or the treatment of flexible the novel vascular diseases subject to indicate of as a subject to has a pharmacological treatment, for example, administering drug, and aim of weakening the novel vascular diseases at least one water. Therefore, the paper, terminology the treatment of intends to a curing and improving the new blood vessel expanding or a disease at least one water. Therefore, the paper, a treatment of a or are opening of pharmaceutical composition, for treating or preventing eye novel vascular diseases.

[0113] A patient a expressed any animal. Terminology of animal comprising a mammal, comprising, and not limited to the other human and primates. The terminology farther comprising domestic animal, to cattle, pig, sheep, horse, three and cat.

[0114] A PDGF shape or a platelet-derived growth factor preparation of capable of affect the blood vessel with or a process for mammal platelet-derived growth factor. The paper, terminology a PDGF comprising a PDGF each pair of hypotypes, card further chart I (A) and (B)) comprises PDGF-B and PDGF-A (card further chart I (C) and (D)). The evening divination, wherein the paper, terminology a PDGF expressed of the PDGF- related for factor, which PDGF-C and PDGF-D, they with an effect through the homologous PDGF receptor, the stimulating blood vessel is or a process for. Specifically, terminology a PDGF a expressed of any member of growth factor type, and (i) (card is between 3 (A) and (B)) or PDGFR-A (card is between 3 (C) and (D) the PDGF receptor connected with such as PDGFR-B; (II) and activating VEGF receptor related tyrosine activating enzyme; activenessA heel (iii), thus affects vascular with or a process for. The paper, terminology a PDGF two generally expressed of the driving member of growth factor type, comprising a through up and activating the platelet-derived growth factor battery surface receptor for reactivity type battery (i.e., PDGFR) connected with the DNA synthesis and mitosis for. Special biological function according PDGFs can realize comprising, for example: Directional mobile battery (chemotaxy) and activating battery; Phospholipase activating; Decorative phospho-lipin acid support phaseomannite turning and prostaglandin, metabolismStimulation reactivity battery collagen and collagenase body; The cellular metabolism activeness, comprising a base body, battery factor production, and lipoprotein absorption; The gap PDGF receptor the battery indirect induction of hyperphasia response; with effective vasoconstriction activeness. Terminology a PDGF a expressed comprising a PDGF the polypeptides and corresponding to PDGF and coding gene or nucleic acid.

[0115] A PDGF-A expressed of the PDGF A chain polypeptide and corresponding coding gene or a nucleic acid.

[0116] A PDGF-B expressed of the PDGF B chain polypeptide and corresponding coding gene or a nucleic acid.

[0117] Of VEGF shape or a vascular endothelial growth factor preparation of capable of affect the blood vessel with or a process for mammal vascular endothelial growth factor. The paper, terminology of VEGF of (a dart and blood vessel through factor (VPF) and VEGF-A) comprises VEGF each pair of hypotypes (card is between 2 (A) and (B)), they are for example, the alternative spliced production of VEGF-A/VPF gene, comprising VEGF121, VEGF165 and VEGF189. Moreover, the paper, terminology of VEGF a expressed of the VEGF- related for factor, to PIGF (placenta growth factor), VEGF-B, VEGF-C, VEGF-D and VEGF-E, they with an effect through the homologous VETO receptor, the stimulating blood vessel with or a process for. Specifically, terminology of VEGF a expressed of any member of growth factor type, comprising (1 )of the right card further gaskets with such as VEGFR-I (4 Flt-I (A) and (B)), VEGFR-2 (KDR/Flk-1) (card is between 4 (C) and (D)) or the VEGFR-3 (FLT-4) VEGF receptor connected; (II) and activating VEGF receptor associated with ammonia

1 Acid activating enzyme; activenessA heel (iii), thus affects vascular with or a process for. Terminology of VEGF a intends to a of VEGF the polypeptides and corresponding of VEGF and coding gene or nucleic acid.

[0118] On the PDGF antagonists of said a part for or completely, or inhibiting reagent for PDGF activeness or production. The PDGF antagonists for directly or reduced or pressed special PDGF indirectly, to PDGF-B. Moreover, with a antagonists to one of the PDGF antagonists of a comprising is able to affect the PDGF ligand or wherein homologous receptor, thus or for inhibiting PDGF- related receptor signal a reagent. Therefore, wherein two ends of the PDGF antagonists of such comprising, for example: The target-type of the PDGF nucleic acid the antisense, ribozyme or the composition RNAi; Sectional area of pdgf for body, Resists and pdgf antibody or soluble PDGF receptor bait, comprising for preventing and PDGF wherein homologous receptor connected; The target-type of the homologous PDGF receptor (PDGFR) nucleic acid the antisense, ribozyme or the composition RNAi; Sectional area of pdgfr for body or sectional area of pdgfr antibody, comprising a binding the homologous PDGFR receptor; With PDGFR tyrosine kinase inhibitors.

[0119] On the VEGF antagonists of said a part for or or inhibiting reagent for VEGF activeness or production of. The VEGF antagonists for directly or or for suppressing special VEGF indirectly, to VEGF165. Moreover, with a antagonists to one of the VEGF antagonists of a comprising is able to affect the VEGF ligand or wherein homologous receptor, thus or for inhibiting VEGF- related receptor signal a reagent. Therefore, wherein two ends of the VEGF antagonists of such comprising, for example: The boot at the VEGF nucleic acid the antisense, ribozyme or the composition RNAi; Sectional area of vegf for body, Resists and vegf antibody or soluble VEGF receptor bait, comprising for preventing VEGF and homologous receptor connected; The target-type of the homologous VEGF receptor (VEGFR) nucleic acid the antisense, ribozyme or the composition RNAi; The light of the homologous VEGFR receptor sectional area of vegfr for body or sectional area of vegfr antibody; With VEGFR tyrosine kinase inhibitors.

[0120] A novel suppressing vascular diseases and outer for treating or preventing form a effective amount of sufficiently antagonists of the invention combined disease or the water can of. For implementing the invention is used for medical treatment of effective amount of according to the alleviation groove and novel vascular diseases the bumps, a weighing and the general health conditions anatomy part and patient change of the novel vascular diseases caused or wound and novel vascular diseases condition active antagonist. Chinese, the doctor or a veterinarian at the proper amount and dosage the. The wherein there is dart and used for the novel vascular diseases and foaming.

[0121] By the following detail and claim book, wherein understand the invention the treatment and merits.

[0122] X-RAY comprising a raised form a preparation with peptide X-RAY amino acid sequence, polypeptides, one or more amino acid residues with variable. The raised formed with a conservative and a steering; the amino acid of substituted provided with structure or a of the (e.g., substituted is leucine with isoleucine). Electric is rarer, the raised form possibly with a non- conservative and a steering (e.g.. substitutes of glycine with tryptophan). Production small change of further comprises a amino acid flaw or the inserting, or are two. The glass according the amino acid residues of substitute, inserted or the flaw and) respectively and biological or the immunology active guide capable of the computer remote of the field knew comprising a to discover, for example, LASERGENE soft (DNASTAR).

[0123] Terminology a raised form, is used for lighting of the polynucleotide sequences, which comprises a gene or wherein coded sequence related polynucleotide sequence. The one of further comprises, for example, a allele) and a film editing) and a species shape or a polymorphism a raised form. The assembling or can be a remarkable identity with the A/D molecules. Natural, the alternative the joint of exon during the mRNA processing, comprising or a smaller of polynucleotides generally. The corresponding comprising a provided with the function, specifically or equipped with the field. Species raised with different polynucleotide in sequence species. Generally, polypeptide of which each other with remarkable amino acid identity. Polymorphism raised is formed between specific species individuals specific gene the steering of polynucleotide sequence.

[0124] Terminology a supporting preparation of capable of transport with the nucleic acid nucleic acid molecules of which is. A type's for supporting the episome, namely, which can prevent nucleic acid outwardly the chromosome duplicates. With a carrier material and/or type independently with a nucleic acid carrier according a connecting. A instruct can be use of dart connected with the gene carrier according type of which is connected with the paper in type of vector. Generally, be used in the recombinant DNA the technical type vector usually) and plasmid to form, comprising expressed the annular of double-bunch DNA ring generally, wherein every supporting formats, they are with the. Each substituting, and plasmid) and a carrier of a exchange use, can the plasmid of the supporting form of the open for. Natural, the invention intends to comprising such are formed with type vector, they served function according equated, and they afterward for the field of male knowledge.

[0125] Combined treating

[0126] The invention bottom part of VEGF and PDGF active special inhibitory effects, use are growth factor antagonists as effective treatment method, for treating to cover the novel vascular disease patient. The PDGF antagonists and alleviation combination of VEGF antagonist, provides for treating eye novel vascular diseases preferably employs any one of antagonists higher treating profit single. The directed toward the fact are two or more of factors with the surface to display the research of remarkable synergism the electric retina endothelial battery system blood vessel, Resists and vegf and sectional area of pdgf the combined function of agent is unexpected (card further Castellon and so on, (2001) Exp. Eye Res. 74: 523-35).

[0127] PDGF and VEGF of the new blood vessel in body, and stimulant substances of the eye growth. Provided is a combined treatment according to suppress PDGF and VEGF biological activeness end is treating or preventing the method of novel vascular diseases.

[0128] Therefore, the invention relates to for treating combined to suppress methods and compositions and new vascular diseases. Specifically, the invention use of the two different intercellular space communications signal guide ways of the vascular battery provided with a effect, namely PDGF and VEGF signal conductive, a novel vascular diseases, to novel eye vascular diseases treating target. The combined method can be used for treating any of a development of eye for newborn vascularization special and marker's ophthalmology disease or conditions, comprising, and not limited, optic nerve plate newborn vascularization and iris newborn vascularization and retina newborn vascularization and choroid newborn vascularization and cornea newborn vascularization and vitreous body newborn vascularization, green cataract, pannus, pterygium, yellow part and dropsy diabetic yellow and dropsy, blood vessel original and retinopathy, retina denaturation, yellow part denaturation, uveitis, retina inflammatory disease and hyperphasia vitreous body retina pathological the. The combined treatment, restraining PDGF (for example PDGF-B) and VEGF (for example VEGF-A the signal conductive antagonists is, preferably independent which are two or more of treatment massage tray with several protrusions, the space for treating effect. Such according although discussed the lower, the described the combined sole of PDGF antagonists and sole VEGF antagonist, further understandable, wherein the sections of multiple types of antagonists possibly is placed.

[0129] According to the invention sectional area of pdgf and sectional area of vegf combined treating can be is independently, or are connected with the therapeutic combinations, and claims a household, medical offices, clinic and hospital waiting or room for hospital. The treating opened on the hospital generally, so the doctor is tightly observe the treatment by, and adjusting of any or. Combined treating duration is fixed by accepting the treatment the bumps and conditions of the patient type disease patient and new vascular diseases end and type and a response to the treatment. Moreover, comprising a development for the novel vascular diseases the risk of (e.g., diabetes) capable of adjusting to therapy, to suppress or the outbreak of delaying to sign. A remarkable advantages of the invention is, the PDGF antagonists and combination of VEGF antagonists for treating novel vascular diseases, such that can employ the lower dose each pair of antagonist, at least one total active antagonists, accordingly claims a production effect of the low toxicity and side effect and a low-cost.

[0130] The combined each of the dosage are corresponding and can be independent control. For example, a antagonists of each day employ three times; the second a of antagonists of each day employ with a. The combined treatment of claims a baffle and circulating form, comprising a silent period section; a patient according could with the plus to recover of any still the unforeseen side effect. The antagonists is prepared by, for the alleviation are two or more of antagonists.

[0131] PDGF and VEGF antagonists target

[0132] PDGF the first from blood platelet decomposition tie, and is determined to exist of the blood serum but not the main growth promoting activeness the blood plasma. First pressing mitosis activeness function of display PDGF on phorocyte, multiple fibroblast and a smooth rib battery, and exists in the neuroglia battery and culture medium. With already appraised two types of homologous PDGF same labour, PDGF A and B, they of independent coding gene (7) is syringe needles and close. 22) chromosome. The main type from blood platelet of the AB heterodimer, although are three or more of the dimers (AA, AB and BB) is naturally exist. The movement, the PDGF dimer is processed wherein the 30kDa secretion protein.

[0133] Already appraised unified the PDGF two types of battery proteins surface of the upper end, α and β (Heldin and so on, (1981) Proc. Natl. Acad. Sci. USA (78): , 3664WilIiams and so on, (1981)Proc. Natl. Acad. Sci. USA (79): 5867). Are two types including five immune globulins one of type extracellular, specifically a cross film field and battery tyrosine activating enzyme, the field of separated by activating enzyme for inserting. The following years multiple, comprising already expounded three or more of PDGF same labour to three or more of receptor dimers (α/α and α/β and β/β) specific. α and receptor homology dimer a binding are three or more of PDGF same labour of the upper end, β and receptor homology dimer of a binding PDGF BB of the upper section, wherein a light of the PDGF AB end lowly about 10 times, and α/β and receptor heterodimer a binding PDGF BB harvester grain opening PDGF AB of the upper section (Westermark & Heldin (1993) Acta Oncologica 32: 101). The specific mode and which is at a- chain is a binding α and receptor, and B- chain capable of the upper section of light of α and β of the receptor Jacchy and ability created.

[0134] Generally, the invention can comprise a or the multiple types of PDGF active reagents. Are PDGF- depressor, or the PDGF antagonists of affect european or the side of the PDGF ligand. Platelet-derived growth factor comprising A- chain (PDGF-A) and B- chain (PDGF-B) homology or heterodimer, they are up two types of related receptor tyrosine kinases, (α) and receptor (PDGFR- (α)) and (β) and receptor (PDGFR- (β)), and wherein two polymerization display functions. Moreover, already appraised the PDGFR compound two types of novel proteinase activating ligands, PDGF-C and PDGF-D (further card cleaning and others, (2000) Nat. Battery. Biol. 2: 302-9; Bergsten and so on, (2001)Nat. Battery. Biol. 3: 512-6; With Uutele and so on, Q001) Hardening 103: 2242-47), „At the PDGFRs different ligand and specific, known PDGFR- (α] (α] a binding PDGF-AA, PDGF-BB, PDGF-AB and PDGF-CC; PDGFR- (β] (β] a binding PDGF-BB and PDGF-DD; And PDGFR- (α] (β) (card further Betsholtz the light of PDGF_AB, PDGF-BB, PDGF_CC and PDGF-DD and so on, Q001) BioEssays 23: 494-507).

[0135] VEGF is secretion the homologous dimer of disulphide are connected, which can selectively the electric endothelial battery multiply, being, and generating matrix degradation enzyme (Conn and so on, (1990) Proc. Natl. Acad. Sci. USA (87): 1323-1327); Ferrara harvester grain opening Henzel (1989) Biochem.Biophys. Res. Commun. 161: 851-858); With P per and so on, (1991) Biochem. Biophys. Res. Commun. 181: 902-906; Unemori and so on, (1992). J Battery. Physiol. 153: 557-562), They form a process for the new blood vessel is. VEGF four base (VEGF-121, VEGF-165 and VEGF-189, VEGF-206 of), the is at the alternative spliced of VEGF gene production (Houck and so on, (1991) Mol. Endocrinol. 5: 1806-1814; Tischer and so on, (1991) J. Biol. Chem. 266: 11947-11954) „Two small form of proliferate, and preferably and two a is mainly located at the battery film, the person is to joint the upper end and. VEGF-165 further can with a heparin and; and richest form. VEGF-121, namely cannot with the pedal form of bleedings, and so on lower end of the VEGF receptor (Gitay-Goren and so on, (1996) J.Biol. Chem. =5519-5523 271) And a lower pressing mitosis for (Keyt and so on, (1996) J. Biol. Chem. 271: 7788-77) Maos. The biologic function of VEGF is (Flt-I and Flk-1/KDR) and two types of tyrosine activating enzyme receptors, wherein every expressions limited via the butt great to the battery for bast derived (de Vries and so on, (1992) with 255: 989-991; Millauer and so on, (1993) battery 72: 835-846; Terman and so on, (1991)Oncogene 6: 519-524). Although two types of the expressions of functional receptors and upper end and can, further chemotaxy of the endothelial battery and promoting the mitosis signal conductive component and a through (Park of the KDR receptor for upper and so on, (1994) J.Biol.Chem.269: 25646-25654; Seetharam and so on, (1995)Oncogene 10: 135-147; Waltenberger and so on, (1994)J. Biol. Chem. 26988-26995). Recently is already bottoms of VEGF gene the single allele (Carmeliet and so on, (1996) and 380: 435-439; Ferrara and so on, (1996) and 380: 439-442) Or Flt-I two alleles (Fong and so on, (1995) And 376: 66-70) Or Flk-I gene (Shalaby and so on, (1995) And 376: 62-66 The mouse body confirms that and VEGF receptor VEGF importance to the blood vessel growth. On each pair of sleeved, discovered vascularization obvious exceptionally with such that the embryo lethal.

[0136] Now known of the redemptive for of tissue oxygen deficit induction of VEGF is a (Levy and so on, (1996) J. Biol. Chem. 2746-2753); Shweiki and so on, (1992) and 359:843-845). Preparation already the research of the external set with high concentration of the angiogenesis retina disease vitreous VEGF body, wherein a non- activity or not one of the vascularization disease and not exist to the high concentration VEGF. Testing the yellow of the operation of the human choroid tissue for shredding and displayed by the high VEGF layer inverse.

[0137] Outside of the cover known endothelial battery specific mitosis original, VEGF of the angiogenesis growth factor cloth, displayed by inducing of the vessel comprising a permeability of macro-molecule to the field of instant and (, therefore, wherein and name of substituted is blood vessel through factor, VPF) (card further Dvorak and so on, (1979) J. Immunol. 122: 166-174; Senger and so on, (1983) With 219: 983-985; Senger and so on, (1986) Res Cancer. 46: 5629-5632) „Colour protein substitute for plasma for blood vessel and permeability of the deposition of gap outside the blood vessel, conveying is to provide with the temporary matrix of endothelial battery mobile form a new blood vessel (Dvorak and so on, (1995) Am. J. Pathol. =1029-1039 146). Permeability is too high truly of the cloth the new blood vessel, comprising a tumour related blood vessel.

[0138] PDGF and VEGF antagonist

[0139] outline

[0140] The invention claims combined treating PDGF and VEGF a novel vascular diseases antagonists (i.e., depressor). Specific PDGF antagonists and VEGF antagonists for the field of male knowledge, and described on the lower part provided. Now or other PDGF antagonists and VEGF antagonists for the technical for of obtain comprising a antibody, the surface body and antisense widowed a body, ribozyme and RNAi composition, they can appraise through the field the guide and instruction of a conventional exercise each substituting and producing, comprising a part below the further claims.

[0141] PDGF antagonist

[0142] Generally, PDGF (e.g.. PDGF-B) and is connected by the method. For example, a obtain is able to suppress multiple types of PDGF antagonists for the PDGF activeness or production, and a wherein a method for the invention. The PDGF antagonists of showing comprising a PDGF nucleic acid or ligand for body, to the described as follows. Moreover, wherein the PDGF antagonists is, for example, Resists and pdgf antibody or connected antibodies. Therefore, a restraining wherein connected with the receptor is disposed with the PDGF molecule of. Moreover, which can such as antisense RNA, ribozyme and RNAi molecule nucleic acid molecules is used as the invention is a nucleic acid layer for inhibiting PDGF type the antagonist. The PDGF antagonists a peptide, protein, a peptide or small organic compound. Moreover, and destroying wherein other signal conductive, for inhibiting the PDGF signal conductive activeness, for example, and use multiple types of small molecular tyrosine activating enzyme suppressant antagonist, comprising a antagonists for is described. The compound or a reagent on the ability of PDGF antagonists role according to the field of known method for determining, and evening divination, a card the following literature, for example, And and so on, (2001) Genes & Dev. 15: 1913-25; Zippel; said (1989) Eur. J Battery. Biol. 50(2): 428-34; With Zwiller; said (1991) Oncogene 6: 219_21.

[0143] The invention farther comprising field the male knowledge's PDGF antagonist, and a wire support, and belongs to the technical a personnel knowledge of any and is equates a reagent. For example, the rotation PDGF inhibiting antibody for the field the male knowledge, for example, described of the simulating patent of 5,976,534,5, 833,986,5, 817,310,5, 882,644,5, 662,904, 5,620,687 and 5,468,468 and PCT WO are 2003/025019, wherein the literature content is punished the article to periphery of the Israeli foot text form. Moreover, the invention comprising benzyl And one 2) and pyrimidine derivatives containing, which is a PDGF antagonist, for example claims a simulating patent of 5,521,184 W02003/01354U and WO 2003/078404, WO 2003/099771, WO 2003/015282 and WO are 2004/05282, wherein the literature is punished the article to periphery of the Israeli foot text form.

[0144] A block the PDGF function small molecular for the field the male knowledge, for example, described of the simulating patent number (6,528,526 PDGFR tyrosine kinase inhibitors), (6,524,347 PDGFR tyrosine kinase inhibitors), (6,482,834 PDGFR tyrosine kinase inhibitors), (6,472,391 PDGFR tyrosine kinase inhibitors), 6,696,434 and 6,331,555,6, 251,905,6, 245,760,6, 207,667,5, 990,141,5, 700,822,5, 618,837 or 5,731,326, wherein the literature content is punished the article to periphery of the Israeli foot text form.

[0145] A block the PDGF function of protein and polypeptide for the field the male knowledge, for example, described of the simulating patent number (6,350,731 PDGF peptide interleukin-11), said 5,952,304, wherein the literature content is punished the article to periphery of the Israeli foot text form.

[0146] For inhibiting EGF and/or PDGF receptor tyrosine kinase uniformly and one with the bicyclic aryl residue and mixing aryl compound for the field the male knowledge, for example, claims for example the simulating patent of 5,476,851,5, 480,883,5, 656,643,5, 795,889 or 6,057,320, wherein the literature content is punished the article to periphery of the Israeli foot text form.

[0147] For restraining the PDGF antisense oligonucleotide for the field the male knowledge, for example, described of the simulating patent of 5,869,462 and 5,821,234, wherein the content of each literature punished the article to periphery of the foot text form.

[0148] For restraining the PDGF surface body (is dart and nucleic acid ligand) is the field the male knowledge, for example, claims for example the simulating patent of 6,582,918,6, 229,002,6, 207,816,5, 668,264,5, 674,685 or 5,723,594, wherein the content of each literature punished the article to periphery of the foot text form.

[0149] The field of male knowledge for restraining PDGF the compound, comprising describes of the simulating patent of 5,238,950,5,418,135,5,674,892,5,693,610,5,700,822,5,700,823,5,728,726, 5,795,910,5,817,310,5,872,218,5,932,580,5,932,602,5,958,959,5,990,141, 6,358,954,6, 537,988 or 6,673,798, wherein the content of each literature punishes the article to periphery of the foot text form.

[0150] VEGF antagonist

[0151] VEGF (e.g.. VEGF-A) are composed of the method. For example, which can comprise multiple types of VEGF antagonists for the VEGF activeness or production, comprising a nucleic acid molecules, the surface body and antisense RNA, ribozyme, the RNAi molecule and VEGF antibody to obtain, and can be used in the invention is the method. The VEGF antagonists of showing comprising a VEGF nucleic acid ligand or suitable body, common to the following described. To VEGF-A special for antagonists of EYE001 (of the dart NX1838), which is a decorative, the PEG surface body, and capable of upper and specific hydrophilic connecting main soluble of VEGF same labour (card, simulating patent of 6,011, 020,6th, 051,698,with 6,147,204). The telescopic body is capable of a positioning of the upper section antibody similar to of VEGF, and arced the VEGF of. The pair of VEGF for suitable body is EYE001, wherein The And Staggered) is formed. Moreover, wherein the VEGF antagonists is, for example, Resists and vegf antibody or connected antibodies. Therefore, a restraining a combined so the VEGF molecule of receptor. Moreover, which can such as antisense RNA, ribozyme and RNAi molecule nucleic acid molecules is used as the invention is a nucleic acid layer inhibiting VEGF type or VACCINE the stable antagonists in method and composition. The VEGF antagonists a peptide, protein, a peptide and small organic compound. For example, which can can also be used as antagonists in light of the VEGF receptor and non- accompany signal conductive active soluble detruncation of VEGF. Moreover, the VEGF signal conductive activeness a through destroying wherein other signal conductive suppressing, for example, is simple multiple types of antagonists, comprising VEGF receptor tyrosine kinase active small molecular depressor, common to the following further claims.

[0152] The compound or a reagent is playing the ability of VEGF antagonists role, a according to multiple types of adjusting method determinations of the field knew comprising a. For example, a of VEGF biological activeness connected on the receptor connected with vascular endothelial battery the vascular permeability. The interaction so the relaxation of the bast connected, wherein consequence is to create of leakage blood vessel liquid. Capable of measuring of the body by vascular leakage of VEGF induction, comprising a rail; the liquid injection Evans and dye of VEGF paraffin to be (Dvorak from leakage guinea of pig vascular system and so on, a Vascular Permeability Factor/Vascular Endothelial Growth Factor, Microvascular HyperpermeabiIity, and AngiogenesisJP (1995) Am. J. Pathol. 146: 1029). , Wherein a method for measuring is similarly for measuring to the antagonists to block VEGF the organism activity ability.

[0153] A for example of blood vessel permeability, VEGF165Q0-30nM) and ΕΥΕ 001 (30 ηΜ _1 μ Μ) or candidate VEGF antagonists exsomatize (ex vivo) premix, and afterward same through a a liquid injection the guinea pig blood barrel and a skin. 30 Minutes of the injection, a injecting part on Evans and leakage dye prevent quantitative according to the adjusting method, which is composed of a three-dimensional analysis system for use computerizes. For inhibiting the VEGF- induction teaching dye is considered is a compound of the vascular minimizing system is a method and composition of the invention for antagonist.

[0154] For determining compound whether the other determination method of VEGF antagonists of the so-called cornea blood vessel with a of. The measuring method; and a VEGF165 (3pmol) methacyrate polymer part to implant the corneal macroporous stroma, to induce the blood vessel growth of the bone non- normal blood vessel cornea. Toilet in the groove through vein employs the candidate VEGF antagonists the rat body, the dosage is lmg/kg, 3mg/kg and 10mg/kg, and ground a disposable or twice, same is 5 days. The treating making disposable, wherein on the micro-photography to periphery individuals' corneas. The new blood vessel of the degree of the cornea tissue cultivating, and candidate compound and their inhibitory actions, wherein the quantitative through the standardized of three-dimensional analysis of photomicrograph afterward. With is phosphoric acid cushion a water (preferably PBQ processing, for inhibiting the compound of the cornea the VEGF- dependent form blood vessel with a considered of the method and composition of the invention for antagonist.

[0155] The mouse model for early maturity retinopathy with appraised the candidate VEGF antagonist. A for example, and placing with and son 9, 8, 8, 7 and 7 mice of the indoor air or oxygen increasing content, and or candidate VEGF antagonists with phosphoric acid cushion a water (PBS) deals by the peritoneum part (e.g.. deals with a lmg/kg, 3mg/kg or the 10mg/kg/of). A toilet, wherein from coming is processed with and mouse comparison each eye's 20 layers histology for the microscope evaluating and a new blood vessel bud counting estimates of the end point, namely a new blood tubular growth through cornea's the film to the vitreous humor the. The raw lower the comparison with processed comprising a decelerator of retina novel vascular system is determined for evaluating for VEGF antagonist.

[0156] The screening method for measuring of the rotary, wherein the human body tumour heterotransplantation to determine the dried candidate VEGF antagonist. The pair of shielding of; a level human tumour xenograft the bare mouse (A673 rhabdomyosarcoma and Wilms tumour) and candidate antagonist VEGF body effect. Toilet with candidate VEGF antagonists processing mouse (e.g.. 10mg/kg, wherein peritoneum same and a telescopic sleeve, comprising develops for defining tumour QOOmg)). Comparison with reagent processing control set. Which is capable of inhibiting the A673 rhabdomyosarcoma growth and Wilms tumour candidate compound of the opposite comparison is considered of the method and composition of the invention for antagonist.

[0157] Determining the VEGF antagonists activeness and method for the field the male knowledge, and further claims a a wire.

[0158] Are of the invention farther comprising field the male knowledge's VEGF antagonist, and a support, with common technique for knowledge range of any and is equates a reagent. For example, a modified of the VEGF inhibiting antibody for the field the male knowledge, for example, described of the simulating patent of 6,524,583,6, 451,764 (the VRP antibody), 6,448,077,6, 416,758 and 6,403,088, 6,383,484, 6,342,221 (sectional area of vegf (is directed toward VEGF-D) (is directed toward VEGF-C the antibody), 6,342,219,6, 331,301 (the VEGF-B antibody) and 5,730,977, and PCT of claims WO 96/30046, WO 97/44453 and WO are 98/45331, wherein the literature content is punished the article to periphery of the Israeli foot text form.

[0159] VEGF receptor for antibody the field the male knowledge, for example, claims for example the simulating patent two similarly 5,840,301 and 5,874,542,5, 955,311,6, 365,157 and PCT of claims WO are 04/003211, wherein the literature content is punished the article to periphery of the Israeli foot text form.

[0160] A for example inhibiting VEGFR- related tyrosine activating enzyme activeness to block VEGF the small molecular of function for the field the male knowledge, for example, described of the simulating patent of 6,514,971 and 6,448,277,6,414, 148,6, 362,336 and 6,291,455,6, 284,751,6, 177,401,6071, 921 and 6001,885 (a substance of inhibitor of VEGF type) are; the content of each literature punished the article to periphery of the foot text form.

[0161] A block VEGF protein and polypeptides of function for the field the male knowledge, for example, described of the simulating patent of 6,576,608, 6,559,126, 6,541,008, 6,515,105 and 6,383,486 (VEGF receptor inducing), (6,375,929 to VEGF receptor in), (6,361,946 VEFG peptide interleukin-11 depressor), (6,348,333 to VEGF receptor in), (6,559,126 is capable of light of VEGF, and blocked with VEGFR and polypeptides), 6,100,071 (VEGF receptor inducing) and are 5,952,199; the content of each literature punished the article to periphery of the foot text form.

[0162] A lead and/or the VEGFR gene expression and/or active VACCINE VEGF to disturbs (RNAi) in a nucleic acid (siNA) and short in RNA (siRNA) and a including RNA (dsRNA), a RNA (miRNA) and a hairpin RNA (shRNA) is the field the male knowledge, for example, claims a PCT of claims WO are 03/070910, wherein the literature the content to punish the article to periphery of the foot text form.

[0163] For restraining the VEGF antisense oligonucleotide for the field the male knowledge, for example, described, for example, the simulating patent of 5,611,135,5, 814,620,6, 399,586,6, 410,322 or 6,291,667, wherein the content of each literature punished the article to periphery of the foot text form.

[0164] For restraining the VEGF surface body (is dart and nucleic acid ligand) is the field the male knowledge, for example, described, for example, the simulating patent of 6,762,290,6, 426,335,6, 168,778,6, 051,698 or 5,859,228, wherein the content of each literature punished the article to periphery of the foot text form.

[0165] antibody antagonist

[0166] The invention comprises a modified of PDGF and VEGF and their homologous receptor PDGFR and VEGFR antagonists antibodies. The invention antibody antagonists of the block ligand and homologous receptor and. Therefore, PDGF antagonists antibody of the invention comprises a modified of PDGF and PDGFR target's antibodies.

[0167] Antagonists antibody of the invention comprises inhibiting monoclonal antibodies. Monoclonal antibody, wherein or connected, comprising an immune globulin types, to IgM, IgG, IgD, IgE, IgA or wherein subtype, to IgG subtype or wherein agent. IgG and subtype are provided, to IgG1, IgG2, IgG2a, IgG2b, IgG3 or IgGM. The IgG subtype IgGl7kappa ^P IgG2ivkapp is taken for a preferred embodiment. The fragments capable of mention is detruncation or with decorative the antibody connected, has one or two antigens co-grinding combining sites, and displayed by a modified of mammal PDGF or VEGF (or parallel homologous receptor the upper connector and ink activeness, to the partial antibody with is equal to the antibody the combining site, and is made of light chain and multiple bond, to Fv, Fab or F ab' (2), crushed or single stranded connected. Of double-bunch fragments of detruncation, to Fv, Fab or F (ab `2) is special for. For example, wherein the connected with the enzyme promoting method by using such as papayin or pepsin enzyme removing to the antibody the Fc part, or are connected with the genetics operation to obtain to the antibody gene by the chemical oxidation. Similarly adopts and is convenient to use genetics operation, the fragments of non- detruncation. The sectional area of pdgf or the VEGF antibody or wherein connected with the independent or are made of the agent.

[0168] The novel antibody and antibody connected, every agent or derivative or VEGF (or parallel the homologous receptor) is PDGF and hydrophilic advantageously the ι χ KT7M-I X-RAY 10-1 ), or 1 * IO-8M-I * 10 of each END; a 1X1 (T9M-5X1 (T1CIM the range.

[0169] For example, the antibody gene for genetic manipulation separable from the hybridoma battery of the groove of the technical field for male knowledge, for the end, and antibody producing battery, and small and light - density is enough, and known arranged from the battery separation mRNA, comprising fixes guanidine thiocyanate decomposition battery, and sodium acetate acidification, and phenol and chloroform/isoamyl alcohol extract, and isopropyl alcohol part, and realizes with the ethyl alcohol washing. Toilet with mRNA fixes counter duplication enzymic body cDNA. Comprehensive cDNA can directly or; the genetic manipulation inserted in the proper animal, a biofilm, or a virus vector afterward, and expressed of the proper a biological, wherein genetic manipulation for example, the site-directed mutagenesis, inducted the inserting, conversion, flaw or the base group or. With bacteria or for supporting and pBR322, pUC18/19, pACYC184, λ or the μ for supporting, a cloning to the gene, and at such as backwoods coli (TRACK. coli) bacteria or such as beer sugar for (Saccharomyces cerevisiae) and for expressed.

[0170] The invention also relates is able to synthesize PDGF or the VEGF antibody battery. The battery comprising according to the upper method of transformed the animal, mushroom, the bacterial cell or the battery for. They advantageously of the hybridoma cells or three lump body (trioma) cells, usually is a battery hybridoma. For example, wherein the hybridoma battery through the known groove is from PDGF or VEGF (or parallel homologous receptor the animal production of immunity, and are parallel antibody producing the B cell, screen are cells' PDGF or the VEGF- and antibody, and afterward which are with cells, for example, the human or the animal, for example, the mouse myeloma battery and human lymph metrocyte or the heterogeneous hybridoma (cytomixis card, for example, Koehler and so on, 197^Nature (256:496), or fixes the proper viral infections the battery, and aim of producing immortalization battery line. Is provided with the battery hybridoma line of the fusion production, and mouse hybridoma battery wire is special for. The invention's hybridoma battery line for of the IgG and with antibodies. The mAb antibody of the invention can of the upper section is connected, and for or the biological of a PDGF or VEGF (e.g.. for) activeness.

[0171] The invention farther comprising a sectional area of pdgf or the VEGF antibody's derivatives, they retained parallel PDGF or VEGF- inhibiting activeness, be changed and they served and medicine reagent related a or other multiple types of treatment, for example, potency of blood serum stability or production. The sectional area of pdgf or the example of VEGF antibody derivative comprises a peptide, a peptide use of rods of light of the is provided with P tidomimetics) of the antibody antigens, a or antibody and antibody of connected or peptide liquid carrier connected with fixing, the carrier to polyethylene glycol, glass, synthetic polymer, to polyacrylamide, polystyrene, polypropylene or polyethylene natural polymer, to cellulose, Sepharose or agarose, or and enzyme, or with radioactivity or nonradioactive mark, to 咕, 123I, 125I, 131I, 32P, 35S, 14C, 51Cr, 36Cl, 57C0, MFe and 59Fe, 9°y, 99mTC and 75k clamp or andFluorescence/antibody, fragments or peptide chemiluminescence mark covalent connected; the mark to rhodamine, fluorescein, isothiocyanate, erythrosin protein, and algae protein, fluorescent containing, metal chelate, avidin and a chain claims avidin or idrabiotaparinux.

[0172] The novel antibody and antibody connected, wherein agent and derivative can directly, dryness and back, for example freeze-drying, wherein adhered is attached to the upper supporting or and auxiliary material configurations is used for activeness is used for producing agent. A for example of active and one-third material capable of mention of antibodies other, with a microorganism or inhibiting motion of microorganism the rotation microbial movable material, generally to the antibiotics or the sulfanilamide, the rotation tumour agent, water; one and a water, thereof; and fat, waxing, the inert intermedia or conventions is used for arranged outside the stomach the other material of product, amino acid, thickening agent or sugar. The agent can be for the control of diseases, and can be used for controlling the novel eye vascular diseases and a AMD and diabetic retinopathy disease.

[0173] The novel antibody and antibody connected, wherein agent or derivative can directly or for treating or the diagnostic with the solid or the liquid carrier, enzyme, with radioactivity, or the nonradioactive mark or/chemiluminescence mark and a fluorescence.

[0174] The invention's of PDGF or the VEGF monoclonal antibody to obtain through the field known any method. For example, for PDGF or VEGF (or parallel homologous receptors) prevent immunity to a mammal. Purifying and PDGF and VEGF capable of changing channel obtain (e.g., a Battery Sciences, Norwood, ΜΑ, and other supplier) purchases). Moreover, the human PDGF or VEGF (or parallel homologous receptors) of the placenta for tissue arranged head. For producing sectional one of PDGF or the VEGF antibody's mammal is unzoned, and can be the primate and rodentia (for example mouse, rat or rabbit), cattle, sheep, goat or the dog.

[0175] Toilet, the antibody producing battery, the spleen and take out from the animal immunity body, and a myeloma cytomixis. The myeloma cell to know comprising for the field of claims (e.g., capable of p3x63-Ag8-653, NS-0, NS-l or the P3U1 battery). The cytomixis operation of any common method of male knowledge angle through the field.

[0176] A connecting the battery and cytomixis operation with convex of the HAT the culture medium afterward, a applies hybridoma. A screen is used for generating the rotation of monoclonal antibodies hybridoma. For example, the side of the case capable is conducted with sandwiched enzyme through immunosorbent assay (ELISA) or production method, made, a monoclonal antibody is produced with fixing of PDGF or VEGF (or parallel homologous receptor) hole and. The pair of sleeved, capable to the immune globulin special antibody for improving piece and second antibody, wherein antibody with enzyme, to peroxidase, alkaline phosphatase, glucose oxidase and β - half d- for glycosidase and equal is marked. A through letting of the marker enzyme with the reaction with the substrate, and measuring the colour of production to redirect teat dip detection to the mark. And the substrate, can produce 3,3) are aminobenzidines and 2,2 of diamino double neighbors to connect the anisidines and 4) chloro-naphthol, 4) amino quinizines; the phenylene diamines and equal.

[0177] By the operation, a screen is used for generating sectional one of PDGF or VEGF antibody hybridoma. A through the hybridoma of folium artemisiae limiting dilution method or the soft components method clone cleaning. Which the light, capable of comprising a blood serum or discontinuous respectively connected with the clone comprising a culture medium of culture of blood serum the hybridoma, or a vaccinate of the mouse abdominal cavity, and parts of the ascites, can thus obtain the block clones the hybridomas.

[0178] Toilet from the selecting sectional one of PDGF or the VEGF monoclonal antibody the switch has prevented corresponding ligand/receptor to the connecting and activating (e.g., the battery and according PDGF or the VEGF of system (see the claims text))The ability monoclonal antibody of a ring is detachably and use. Which the antibody to block receptor ligand/connected and activating, the method of monoclonal antibodies of experiment with weaken or capable of PDGF or VEGF PDGF or the VEGF active ability. Frame is, wherein the monoclonal antibody of specific respectively and/or disturb of PDGF or VEGF (or parallel homologous receptor the key combining site.

[0179] The invention's monoclonal antibodies farther comprising through the hybrid and to antibody of the following method of: The film editing sectional area of pdgf or VEGF antibody variable (comprising high of) for and for (e.g., a humanized of antibodies), or a chain and multiple bond, or is from a species the chain on the chain from one pair of species, or blended body and heterologous protein, wherein the first derived species or the immune globulin type or the subtype name, and antibody connected [for example, Fab and (F) Ab 2* Fv), a long and they with a biological activeness of hope on the circuit. Card (further for example, simulating patent of 4,816,567 and Mage&Lamoyi; a Monoclonal Antibody Producing Techniques and Applications, PP. 79-97 (Marcel Dekker, Inc.), Novel York (1987)).

[0180] Therefore, terminology of a monoclonal antibody expressed and is prepared from antibody basically are group, and is not understood is formed by one specific method for producing antibodies. For example, can be used for monoclonal antibodies in the invention is fixed at bottom of Kohler & Milstein, And 256: 495(1975) The hybridoma preparation method of description, or a through the recombinant DNA method (simulating patent of 4,816,567) preparation. A monoclonal antibody and further separable of the technical preparation bacteriophage library with the following literature claims from the use of: For example, McCafferty and so on, And 348: 552) 554(1990).

[0181] Inhuman (e.g., comprising) and an antibody humanized the form of a special the immune globulin and immune globulin chain or wherein connected (for example the other antigens of Fv, Fab, Fab', F Ab (2) or antibody with the back) sequence, comprising a minimal sequence of inhuman immune globulin. The majority of humanized antibody of a person immune globulin (receptor antibody), complementary determining area (CDRs) the residue from receptor antibody component of the inhuman species (donor antibody) to the mouse, rat or rabbit's CDRs residue substitutes, comprising a specific the end, and ability for steps. A plurality situations, the human immune globulin's Fv frame is provided FR) residue substituted are made of inhuman corresponding FR residue. Moreover, humanized antibody and comprising two not exist of the receptor antibody, and not exist of input CDR or the residue-filter in sequence FR. For the upper modified to has further excellent and a of optimized antibodies. Generally, humanized antibody basically comprises at least one additional, the entirety of usual two types of variable, field, CDR is completely or basically completely is equal to the inhuman immune globulin CDR area, and FR residue completely or basically completely is equal to and immune globulin consensus sequence FR residue. The humanized antibody superiorly farther comprising an immune globulin constant area (Fe) at least one part, usually of the person immune globulin constant area.

[0182] A method of humanized antibody producing non-human knew comprising for the field. Generally, a tape from the inhuman origin one or more amino acid residue to an iontophorectic humanized antibodies. The inhuman amino acid residue usually is dart and input of residue, comprising usually component of the input a variable field. Humanized basically capable of carrying (Jones according to a method for Winter and colleague and so on, (1986) Nature321: 522-525; Riechmann and so on, (1988) and 332: 323-327; Harvester grain opening Verhoeyen and so on, (1988) With 239: 1534-1536), Substitutes of the human antibody with rodentia (3 )or Rs (3 )R sequences the corresponding sequence. Therefore, wherein two humanized of antibodies is a chimeric antibody; a already of substituted from the corresponding sequence of inhuman species display of brushing less than the sequence of human variable field. The exercise, the humanized antibody usually is the human antibody; multiple (3 )R residues, and can multiple FR residues component of the rodentia antibody production of the residue-filter to substitute.

[0183] On the light chain and side includes a variable for is used for preparing the humanized antibody is weakening the antigenicity is very substances. According to so-called the optimum matching the method, a variable sequence for rodentia of antibodies to known external variable the field sequence's library whole screen. Toilet served and humanized antibody with the rodentia sequence closest external sequence of frame (FR) (Sims and so on, (1993) J. Immunol., 151: , 2296with Chothia and Lesk(1987) J. Mol. Biol., 196: 901). The pair of method or multiple bond specific subtype of human antibody consensus sequences specific skeletons of light chain. The same framework be used in multiple types of different humanized antibodies (Carter and so on, (1992) Proc. Natl. Acad. Sci. (USA),89: , 4285with Presta and so on, (1993) J. Immnol., 151: 2623).

[0184] Moreover, the importantly antibody humanized, retention's upper end to antigens, and convenient biological characteristics. Are orderly connected to the field, according to a method, and analysis of the base sequences and concepts humanized the method of product for base and three-dimensional model of humanized sequence the preparation humanized antibodies. The three-dimensional immune globulin model can generally obtain, and is familiar for the technical field of personnel. A obtain is explaining and demonstrating while candidate immune globulin sequence the computer remote of the three three-dimensional conformation structure. A for office the rotary to permit and related residue's function of candidate immune globulin sequence display function, namely, and a affect the candidate immune globulin and antigens and residue of ability. Water, and a single entries the switch and combined FR residue from the consensus sequence, thus by the antibody the utility model according, enhancing to target antigens hydrophilic. Generally, CDR residue and is mainly participates of the straight antigens and.

[0185] The invention farther comprising a modified of PDGF or VEGF human monoclonal antibodies. The antibody is connected with the hybridoma preparation method thereof. Already described for producing of monoclonal antibodies human myeloma and mouse one of heterogeneous myeloma battery wire, for example, card further Kozbor (1984) J. Immunol., 133,3001; Brodeur, said that Monoclonal Antibody Producing Techniques and Applications, PP.51-63 (Marcel Dekker, Inc., Novel York, 1987); Harvester grain opening Boerner and so on, (1991) J. Immunol., 147: 86_95.

[0186] Possibly now generating gene animal (e.g., comprising), composed of an immune tank, a without of the premise of the endogene immune globulin with a of human spectra antibodies. For example, already discovered that wherein and a of the sudden the mouse the antibody includes multiple connecting part (JH) gene homozygous flaw, such cover and endogene antibody with meat. Plant external is an immune globulin gene production transfer to planting is a compositions comprising body; the motor and is attacked of the antigens generating human antibody (card, for example, Jakobovits and so on, (1993) Proc. Natl. Acad. Sci. (USA), 90: 2551 Jakobovits and so on, (1993), And 362: 255-258; Harvester grain opening Bruggermann and so on, (1993) Year the Immuno. , 7: 33).

[0187]Moreover, wherein bacteriophage rotary technology (McCafferty and so on, (1990), And 348: 552-553) For generating human antibody and antibody connected by donor immune globulin variable (V) for gene spectrum resuscitation according from coming and is not crossed (the related summary a card further for example, Johnson and so on, (1993) On Opinion of Biological Structure. 3: 564-571). The multiple origins of V- gene connected can be used for bacteriophage display. For example, Clackson and equal ((1991), And 352: 624-628) From coming in a crossed and mouse spleen's triangular small gene and library combined with separated multiply sectional P and alkone antibody array. A through from human donor V gene spectrum of and is not crossed, and a basically and technical separating is as follows claims a modified of the multiple antigens arrays (comprising of antigens) antibody =Marks, (1991) J. Mol. Biol., 222: 581-597, Or Griffith and so on, (1993) EMBO. J, 12: 725-734).

[0188] The innate immune response; the antibody gene of the high in accumulation of bumps (somatic With Said). A change for leading possibly with the upper end, displayed by the upper end surface immumoglobulin the B cell is covered and split is arranged afterward antigens the. The technology of simulate the pair of natural and (card Mark is fixes the dart and a chain to and so on, (199) use with. Technol., 10: 779-783). The method, and bacteriophage demonstrated by a triangular for gene of the first of human antibody's section of fixes never and a piece of the donor of which the raised with spectrum with small periphery (and substituting) is the multiple bond and light chain V area gene in turn improving. The technology of a hydrophilic the antibody and antibody connected in nM range. Waterhouse and equal already described for preparing comprising a bacteriophage antibody spectrum (strategy (1993) Nucl. Acids Res. , 21: 2265-2266) „

[0189] Further capable of generating from the rodentia antibody the gene to the human antibody; the human antibody to a trench end and specific to the end rodentia antibodies. According to the method, which is a dart of mimotopes in mark; and bacteriophage demonstrated the rodentia antibody's and compounding or the light chain is V for gene the technical by substituted are made of human V for gene spectrum, thus provided with rodentia of human chimaera. The switch antigens, such that is able to restore the functional antigen combining site the separating of human variable field, particularly control mimotopes mark (a) are of gametophyte. The process of redundant, and aim of substituting is used for rodentia V field shell, the human antibody (see the PCT WO 93/06213,1993 years on April 1) publicly. A turner knife for rodentia antibody traditional humanized is different CDR through hole, the technical claims a of human antibody, comprising without the frame or the CDR residue from rodentia.

[0190] Telescopic body antagonist

[0191] The invention claims a modified of PDGF and/or VEGF (or parallel homologous receptors) suitable body antagonist. Telescopic body; the dart and a nucleic acid ligand, a nucleic acid for and staggered with small, comprising a binding, and generally the antagonism (i.e. used, the target of preselection.

[0192] For body is of producing any known preparation method of widowed a body or oligonucleotide. Multiple synthesizing method for the field of male knowledge. For example, comprising residual purine ribonucleotide, and is appropriately 3 ' - end, to the residue-filter triterpenes by conversion (Ortigao, Antisense Mounted and Development, 2: 129-146 (1992) Or) is located at 3 )and is di-sulfide generation of phosphate keys for preventing chinese 3) is 2 circumscribing according nuclease degrades ' -0) allyls with decorative the widowed a body, which is solid the β and cyanogen ethyl Asia isophosphoramide gametocide method (Sinha and so on, Nucleic Acids Res. , 12: 4539) 4557(1984)) On the DNA/RNA combiner, wherein any of obtain by changing channel synthesizes. A method of the ribonucleotide 2 ' -0) uncle butyl further silyl (TBDMS) and strategies (Usman and so on, J. Am. Chem. Soc., 109: 7845-7 Magic 4(1987)), and is placed 3 ' _0 each phosphoramidite prepared by changing channel. Moreover, the amino methyl polystyrene and used for supporting the material, the is elastic lock wherein the handle (McCoIlum and Andrus (1991) Tetrahedral. Lett, 32: 4069-4072). A during the synthesis through the made of changing channel of the fluorescein phosphoramidite is prepared, the fluorescein added to a RNA 5 ' - end. Generally, a soybean for body widowed a body through the adjusting the VACCINE. When the making device, and lower 55°C is strong ammonia water in the small of sealing and ethyl alcohol (3: lv/v) for processing method 8 removing and alkaline unstable protection groups and bottom. The ethyl alcohol for inhibiting 2 ' - o-tbdms premature removing of and bottom, the group and bottom and falling of the basic condition of protecting; the ribonucleotide position of the steps of the obvious decomposition chain (Usman and so on, (1987) J. Am. Chem. Soc., 109: 7845-7854). The freeze-drying,/triethylamine /N- methyl pyrrolidone's mixture of 60°C a to the TBDMS protection with triethylamine three hydrofluorides the widowed a body 2 hours, comprising a fast and removing the silyl protection groups and bottom (card further Wincott of the ink effectively improve and so on, (19%) NuCleiC Acids Res. , 23: 26774684). And a precipitate the widowed a body with the butylacohol of protecting, according to Cathala and Brunei method for ((1990) Nucleic Acids. Res, 18: 201). The purification a through denaturing polyacrylamide gel electrophoresis or through ionic or HPLC (Sproat and so on, (1995) Nucleosides and Nucleotides, 14: 255-273) And a combination of opposition HPLC is conducted. for the battery, and precipitates the widowed a body of the soybean with replaceable usage of salt with the sodium perchlorate of the acetone. Toilet for small disposable filtering gel column to remove grains buffer the residual salt, wherein the column by changing channel to obtain. And the tail, wherein by the matrix auxiliary laser desorption mass-spectrometric method (Pieles and so on, (1993) Nucleic Acids Res. , 21: 3191-3196) And a nucleoside base group and composition for office separating the authenticity of widowed a body.

[0193] When the invention subunit can be is used for promoting enzyme the operation, for body is in particular can promote the production method of enzyme. For example, and preparing RNA molecule by external polymerase VACCINE T7 reaction. The molecule further through is able to the outer T7 bacteria strain or the battery and preparation, afterward from the battery separation. Heat-insulated to the following discussed that for body is in particular further capable of the board and promoter expresses the battery is.

[0194] The telescopic body, is similar to the nucleic acid molecules of the invention, further - comprising of the modification of nucleotide. The issue of the diagnostic or therapeutic use of nucleic acid abnormal surface with a di-phosphate, and form oligonucleotide the body fluid of displaying to be the effect of possibly and lyoenzyme a battery, to the inner contact nuclease and circumscribing nuclease degradable fast. Can be a plurality of modifications to the nucleic acid ligand, to enhance nucleic acid ligand body stability, or a reinforcing or lead the nucleic acid ligand section (card further for example, simulating patented multiple claim 5,660,985, title is a High Hydrophilic Nucleic Acid Ligands A Modified Nucleotides”), wherein the literature is punished the article finger pricking device for.

[0195] The modified of nucleic acid ligand the invention anticipated comprising, and down in limiting to provide the 5-site modified of the gametocide groups and bottom, comprising and a whole with conformed with the electric material, polarizability, includes, the hydrogen layers; the static shape and uncertainty (fluxionality) is a nucleic acid ligand base group or to the nucleic acid ligand. The modified comprises, and down in limiting, 2 ' the number of modified position; the pyrimidine modified of an. 5) position; the purine modified of an. 8) position, beautification and containing outer ring, the substitutions of 4) thio triterpenes by, 5) bromines or 5) iodines of the substituted of uracil; The principal chain modified, the sulfo- phosphate or the compound phosphate modified, the methylation, the uncommon pair of combined, to different base group (isobase) different deoxycytidine (isocytidine) and different guanidine (isoguanidine) is equal. The modified further comprises a ( 3 and 5 of modified, which comprises a cap or the modified with a sugar component. The multiple by plans of the invention, wherein the nucleic acid ligand is a molecule VACCINE, and 2 of pyrimidine residue sugar component ' - F) modified.

[0196] A inducting the pair of modified, and is along the principal phosphate chain beautification and substituted of RNA, wherein considerably reinforcing to the stability of suitable body. Moreover, can carry and multiple types of 5-site modified a nuclear base group (nucleobase strips), wherein a of modified for inhibiting the degradation, and invention shape capable of nucleotide reinforcing shape is placed for or not be. Therefore, at knew the sequence of suitable body, an method for male knowledge connected with the modified or the substituted through the synthesizing method or through the technical field of players as follows described.

[0197] The 5-site modified (nucleotide comprising a conformity with decorative the base group or a decorative nucleoside or a decorative), they exist of the RNA (i.e., A, b, and A socket) and DNA (S 卩, A, b, and A T-SHAPED) table base group and sugar and/or phosphate principal chain of the variation. The range comprises, for example: GnK2'- methoxy guanylic acid), ΑπΚ 2 ) and methoxy adenylate), Cf^2 Of The containing cytidylate), Uf^2'- containing triterpenes by urine acid), Ar (ribose) adenylate. The telescopic body further comprises a cytosine or any cytosine and related base group, comprising 5) methyl cytosines, 4) acetyl cytosines, 3) methyl cytosines, 5) methylol cytosines, 2) 5-8% cytosines and 5) halogenating cytosines (e.g.. 5) fluoro cytosines, 5) bromo cytosines, 5) chloro- cytosines and 5) iodo-benzene cytosines), each 5 propynyl cytosines, 6) azo cytosines, 5) are containing methyl cytosines and N4, N4- connecting ethylidene cytosine (ethanocytosine), 吩 P oxazine deoxycytidine, thiophenylamine deoxycytidine, carbazole deoxycytidine or pyrido- formulation deoxycytidine. The telescopic body further comprises a guanine or any guanine and related base group, comprising 6) methyl guanines, 1) methyl guanines, 2,2) further guanines, 2) methyl guanines, 7) methyl guanines, 2) propyl guanines, 6) propyl and guanines 8) halogenating guanines (e.g.. 8) fluoro guanines, 8) bromo guanines, 8) chloro- guanines and 8) iodo-benzene guanines), 8 of each guanines amino, 8) thiol guanines, 8) sulfo- the guanines, 8) hydroxyl guanines, 7) methyl guanines, 8) connected with compound guanines, 7) denitrogenation guanines or 3) denitrogenation guanines. The telescopic body further comprises a adenine or any adenine and related base group, comprising 6) methyl adenine, N6- isopentennyladenines, N6- methyl adenine, 1) methyl adenine, 2) methyl adenine and 2 of methyl Sulfo- and n6- and isopentennyladenines 8) halogenating adenine (e.g.. 8) fluoro adenine, 8) bromo adenine, 8) chloro- adenine and 8) iodo-benzene adenine), 8 of each adenine amino, 8) thiol adenine, 8) sulfo- the adenine, 8) hydroxyl adenine, 7) methyl adenine and 2) halogenating adenine (e.g.. 2) fluoro adenine, 2) bromo adenine, 2) chloro- adenine and 2) iodo-benzene adenine), 2 of each adenine amino and 8 - comprising a mixing adenine, 7) Denitrogenation adenine or 3) denitrogenation adenine. Farther comprising uracil or any uracil and related base group, comprising 5) halogenating uracils (e.g.. 5) fluoro uracils, 5) bromo uracils, 5) chloro- uracils and 5) iodo-benzene uracils), each 5 (carboxyl group methylol) and uracils 5) ethyloic amino methyl one 2) thiouracils, 5) ethyloic amino methyl uracils, dihydride uracils, 1) methyl moveable uracils and 5 and methoxy amino methyl one 2) thiouracils, 5 ' - methoxycarbonyl methyl uracils, 5 and methoxy and uracils 5) methyl one 2) thiouracils, 2) thiouracils, 4) thiouracils, 5) methyl uracils and uracils one 5) hydroxyacetic of methyl uracils and one 5) hydroxyacetic acids, vacation and uracils 5) methyl one 2) thiouracils, 2) Thiouracils, 3) (3) -3-N-2- Afflicted carboxyl propyl) uracils, 5) methyl amino methyl uracils, 5) propynyl uracils, 6) azo uracils or 4) thiouracils.

[0198] The field of male knowledge's other with the decorative of the base group raised form a, and not limited to the 37C. F.R. § (1.822) P (1) enumerates, for example, 4) acetyl cytidines, 5) (carboxyl group methylol) triterpenes by diaper, 2 ) and methoxy cytidines and 5) ethyloic amino Methyl -2-thioridine, 5) ethyloic amino methyl triterpenes by diaper and dihydride triterpenes by diaper, 2 to -0) methyl moveable uridine and b-D- galactose bottom Q nucleoside (queosine), inosine, N6- isopentene adenosine, methyl adenosine and methyl vacation uridines, 1) methyl guanosines, 1) methyl inosines, 2,2 of each further guanosines, 2) methyl adenosines, 2) methyl guanosines, 3) methyl cytidines, 5) methyl cytidines, N6- methyl adenosines, 7) methyl guanosines, 5) methyl amino methyl triterpenes by diaper and 5 and methoxyAmino methyl one 2) thio triterpenes by b-D- and mannose bottom Q nucleoside (marmosylqueosine) and each 5 methoxycarbonyl methyl urine triterpenes by 5 and methoxy each triterpenes by diaper, 2) methyl sulfo- N6- isopentene adenosines and N- (9-b-D- (ribofuranose is one 2) methyl sulfo- purine _6 each) is ammonia formacyl) threonine and N- (9-b-D- (ribofuranose bottom purine and 6) are) And methyl and ammonia formacyl) threonine and transfusing and triterpenes by 5) hydroxyacetic of methyl and transfusing and triterpenes by 5) hydroxyacetic acids (ν), wybutoxosine and vacation uridines, Q nucleosides, 2) sulfo- and cytidines 5) methyl one 2) thio triterpenes by, 2) thio triterpenes by, 4) thio and triterpenes by 5) methyl urinesAnd triterpenes by N- (9-b-D- (ribofuranose bottom purine and 6) are) ammonia formacyl) threonine, 2 to -0) methyl one 5) methyl triterpenes by diaper, 2 to -0) methyl triterpenes by diaper and wybutosine, 3) (3) afflicted 3 )and carboxyl propyl) urine triterpenes by.

[0199] The farther comprising the modified with the following literature described nuclear base group: Simulating patent of 3,687,808 and 3,687,808,4,845,205,5,130,302,5,134,066,5,175,273,5,367,066,5,432,272, 5,457,187,5,459,255,5,484,908,5,502,177,5,525,711,5,552,540,5,587,469, 5,594,121,5, 596,091,5, 614,617,5, 645,985,5, 830,653,5, 763,588,6, 005,096 and 5,681,941. The field the modified of male knowledge the nucleoside and example of nucleotide sugar principal chain raised form a, and not limited to such raised form, comprising a, for example, 2 'ribose bottom substituting group, to F, SH, SCH3, 0CN and capacitor, Br, CN, CF3, 0CF3, S0CH3, S02, CH3, 0N02, N02, N3, NH2 and 0CH2CH20CH3, (0 CH2) 20N (CH3 2) and 0CH20CH2N (CH3 2) and (0 C1_10 the), (0 C2-10 alkene bottom), (0 C2-10 alkyne bottom), S (C1-10 the), S (C2-10 alkene bottom), S (C2-10 alkyne) is, NH (C1-10 the), NH (C2-10 alkene bottom), NH (C2_10 alkyne bottom) and 0 of compound 0 )and. 2 Of being a ribose bottom substituting group comprises 2) and methoxy O' -0CH3), 2 of the amino C oxygen root O Of 0CH2CH2CH2NH2), 2 ) is allyl O Shape of ch2-ch = CH2), 2 to -0) allyl O of -0-CH2-CH = CH2), 2 of the amino (2) and nh2) and 2 ' - F). (2 ' Each substituting group can be of the azelnidipine sugar (upstream) are or the ribose (downstream) in position.

[0200] For body of the invention can or said support combination comprising a nucleotide and/or the invention is interleukin-11 the claims text of that or is a oligonucleotide analogs thereof. For body of the invention can, wherein not affect the widowed a body and PDGF or VEGF (or parallel homologous receptor) of the connecting function of comprising a nucleotide analogs thereof.

[0201] With multiple technique is suitable for improving or reinforcing nucleic acid ligand's unions to a specific targeting molecule or the other surface of the choices of. A technology, generally is dart and an outer genetics the card (further &ostak (1992) TIBS, 19: 89), A through the switch from any sequence storehouse) is suitable body antagonist. Separable for body nucleic acid molecules storehouse according to claims comprises a left 20 to 40 nucleotides the invariable sequence of the variable sequence side joint. The method of already dart and ligand the evolution of a first to be (Evolution Selective of Ligands of Exponential Enrichment) (SELEX). Preparing the invention is SELEX and related method for antagonist body compositions and methods for the field the male knowledge, and taught with the following literature, for example, the simulating patent are 5,475,096; the title is a Nucleic Acid Ligands), a simulating patent are 5,270,163; the title is a Method for Identifying Nucleic Acid Ligands shape; each literature which is punished the article to periphery of the foot text form. Generally, wherein the SELEX method, and said of VEGF, and PDGF surface body and preparation further claims the following literature, for example, the simulating patented multiple claim 5,668,264,5, 696,249,5, 670,637,5, 674,685,5, 723,594,5, 756,291,5, 811,533 and 5,817,785,5,958,691,6,011,020,6,051,698,6,147,204,6,168,778,6,207,816, 6,229,002,6, 426,335,6, 582,918, the content of each literature which is punished the article finger pricking device.

[0202] Simply speaking, the SELEX method of claims from candidate oligonucleotide cleaning agent, and are the same general the method redundant, a separator gradually and adding with a connecting device of target chose, and aim of obtaining the fact to be connecting end and a big table. From typically comprising a randomization sequence connected a nucleic acid agent to start, wherein the SELEX method comprising letting and lower agent and effect of which are contacted with the target-type, a already specific unified the target-type molecule of the nucleic acid and separation a nucleic acid and connected, dissociated to the nucleic acid and target compound, increases from nucleic acid and nucleic acid of the target compound dissociated, comprising obtaining nucleic acid rich comprising ligand agent, comprising repetitive arranged, separated, dissociates and increasing the cycle-index of the step reached is wider, comprising obtaining with height high specificity hydrophilic nucleic acid ligand directed to the target-type molecules.

[0203] Already to realize multiple types of special goals made of the improved to the base SELEX method. For example, the simulating patent are 5,707,796; the title is a Method for Selective Nucleic Acids on the Substrate of Structure, described with the special structure a nucleic acid molecules for SELEX method and gel electrophoresis combined switch, to the DNA. The simulating patent are 5,763,177; the title is a System Evolution of Ligands of Exponential: FeedingPhotoselection of Nucleic Acid Ligands and A SELEX, described for choosing comprises a light reactivity groups and bottom nucleic acid ligand SELEX to the method, wherein the group and bottom is downward arranged and/or light crosslinking and light and lock target molecules. The simulating patent are 5,580,737; the title is a High-Affinity Nucleic Acid Ligands According Discriminate Between Theophylline and Caffeine, textures: )Is appraising 1 )and method of specific nucleic acid ligand, wherein the ligand of the n deflation the closed related molecule, the method can and staggered peptide; the dart and Counter-SELEX. The simulating patent are 5,567,588; the title is a System Evolution of Ligands of Exponential =Solution Feeding SELEX shape, claims a SELEX to the method thereof, wherein a separating efficient to the target-type molecule with the upper end and lower end oligonucleotide.

[0204] The SELEX method comprises a decorative the invention upper end nucleic acid ligand comprising a dried, wherein a decorative the invention is used to entrust with the ligand the improved, to the improved stability body or improved the delivering. The modified such a and/or phosphate and/or base group of the gametocide substituted on ribose. Method SELEX evaluating comprising decorating the nucleic acid ligand description of nucleotide the simulating patent are 5,660,985; the title is a High Hydrophilic Nucleic Acid Ligands A Modified Nucleotides), the literature claims a of the pyrimidine 5) and 2 ' )in position of the upper end of the modification oligonucleotide of nucleotide derivatives. The simulating patent of 5,580,737, and is described, the high specificity nucleic acid ligand, comprising a or more for ( 2 )amino O Shape of nh2), 2 of one F) and/or 2 to -0) methyl O' and OMe) with decorative nucleotide. Simulating patented multiple claim 08 Λ 64,029, 1994 and 6 B H¢1 22 )! )And a Novel Method for Preparing of Known and Novel 2 of Modified Nucleosides of Intramolecular Nucleophilic Displacement, already, gave is described, comprising a or 2 ) and a decorative pyrimidine oligonucleotide.

[0205] The SELEX method thereof and oligonucleotides of and the non- oligonucleotide function unit combined comprising a oligonucleotide of the applies, to simulating patent of 5,637,459, title is a System Evolution of Ligands of Exponential =Chimeric Feeding SELEX), a simulating patent of 5,683,867, title is a System Evolution of Ligands of Exponential: FeedingBlended SELEX; the. The patent an oligonucleotide with multiple types of shape and treatment, and increasing and duplication the effectively with other molecules steps to a harvester.

[0206] A nucleic acid ligand and lipophilic compound or the non- immunogenic and macromolecular weight compound for SELEX method farther comprising a designate or the medical compound type the diagnosis, such as simulating patent are 6,011,020; the title is a Nucleic Acid Ligands Complexes, wherein the literature to punish the article to periphery of the foot text form.

[0207] The telescopic antagonists body is locked in use of computer technique for use improved. The example of molecule analogous system is CHARMm and QUANTA for Polygen, Corporation (ffaltham, Mass). . CHARMm carrying out the energy minimum and molecular dynamics function. The structure, printed use and analysis QUANTA an molecular structure. QUANTA a construct, beautification and display each other, and analysis molecule performance. Is applied suitably of the measuring, and display RNA and DNA molecule secondary structure.

[0208] And a bottle is provided is a of 5-site modified the functions of suitable body, the is suitable for field PDGF or any are of method of VEGF function, to PDGF the battery according and activeness of.

[0209] The modified or is conducted with the SELEX method. The modified front SELEX method of shell and improvement of parallel to SELEX improves the body stable a nucleic acid ligand of specific. The SELEX method to 2 ' - oh nucleic acid ligand decoration, storing the body stable improved, and is connected with the negative force to the nucleic acid ligand's combining capacity.

[0210] Be used for generating the invention the other of 5-site modified body suitable for the field of claims a personnel the male knowledge. The modified can be; the SELEX method and (front afterward appraised is not decorative the modified of ligand) or through conforming is conducted to the SELEX method of.

[0211] Already discovered is generally for body or the nucleic acid ligand, a spoke the VEGF surface body in of stablest, accordingly worked and 5 of the valve cap and 3 of the valve cap is effective, which can weaken to the susceptivity of circumscribing nuclease, and enhanced the whole stability. Therefore, wherein a preferred embodiment, the invention is generally to for adding body cap, and a spoke Resistance of vegf specifically suitable body and cap, and located at 5 ) is 5 to -5 of reverse nucleoside cap structure and located at 3 ) is 3 to -3 of reverse nucleoside cap structure. Therefore, the invention claims A Resisted and vegf and/or sectional area of pdgf for body, the S 卩, a nucleic acid ligand, and is 5 to 5 of tail ( -5 ) and nucleoside cap, and 3 to 3 of tail ( -3 ) is nucleoside cap.

[0212] A for bodies of special for The invention is suitable vegf body composition, comprising, and not limited to with 5 to their terminals ( -5 ) and 3 to -3 of the invention is a structure composition. The Sectional one hole vegf device suitable body is the RNA surface body and DNA surface body or with mixing (S 卩, RNA and DNA composite) suitable body. The invention are Sectional is suitable vegf body sequence a polynucleotide sequence GAAGAAUUGG (SEQ ID NO: 15); or polynucleotide sequence UUGGACGC (SEQ ID NO: 16); or polynucleotide sequence GUGAAUGC (SEQ ID NO: 17). One of which is provided with an wherein the invention sectional area of vegf for body, and has the following: sequence

[0213] X-5' -5 ' - CGGAAUCA (3) GAAUGCUUAUACAUCCG-3 (-3 ) AND * (SEQ ID NO: 18)

[0214] And each C, A, B and socket expressed the invention deoxycytidine, guanidine, adenine and diaper according triterpenes by with an naturally, or with comprising a decorative when the invention; X-5' -5 to 5 of the uniform cap the invention of the surface body; 3 ( -3 ) And * 3 ') comprises the invention is arranged on the surface body; And other nucleotides or a decorative the invention adopts 5) in order to -3 of di-phosphate and keying. The multiple by plans, and a sectional area of the vegf each invention is suitable body with single 2 ' ribose of substituted, to _0H (which is a RNA (RNAs) adjusting), or the pallet (which is a DNA (DNAs) adjusting). The other by plans, 2 ' ribose bottom position 0 seeped (_1|The), 0 seeps (_1|Alkene bottom), F, R3 or the NH2 substituting group substitutes.

[0215] A plurality of non- definition, 5 to -5 of the cap sectional area of vegf for possibly body has the following: structure

[0216] Td-5' -5' _CfGmGmArArUfCfAmGmUfGmAmAmUfGmCfUfUfAmUfAmCfAmUfCfCfGm3' _3' -Td

[0217] (SEQ ID NO: 19) Is, a Gm a represent 2 ) and methoxy guanylic acid, a Am a represent 2 ) and methoxy adenylate, a C/a represent 2) for containing cytidylate, a U/'represent 2 `the containing uridylic acid, a K of represent ribose adenylate, and a Td a represent ribodesose thymidylic acid.

[0218] trans-, ribozyme and DNA zyme antagonist

[0219] The target-type a PDGF and VEGF trans- oligonucleotide and ribozyme; a corresponding PDGF or the VEGF mRNs degradation from protein translations or through which the targets of said messenger RNAs through the inhibit realizing inhibitory effects of PDGF/VEGF. Nucleic acid providing is PDGF and VEGF- target for designing and synthesizing PDGF and for sequence of VEGF ribozyme and trans- oligonucleotide. Design and a method for increasing pear fruit shelf life capable of synthetic trans- oligonucleotide and ribozyme for the field male knowledge. The invention claims other directors.

[0220] The oligonucleotide of the is specific and effective mRNA- a target (trans- ODNs) and problems of ribozyme and trans- appraise the trans- pair of target mRNA of the position (a folding part self- pair secondary structure). Computer assistance's VACCINE prediction pair accessibility algorithm and a molecule screening combined, which can prepared is directed toward the majority of mRNA target's collection and effective ribozyme and/or the trans- oligonucleotide. The fact, already claims a pair of confirm) RNA molecules or accessibility the method for increasing pear fruit shelf life capable of ribozyme subjected to the trans-. A a method for increasing pear fruit shelf life fragrance use in vitro screen the measuring, use and far as with multiple trans- widowed poly well (card further Monia and so on, (1996) and Med., 2: 668-675; With Milner and so on, (1997) and Biotechnol., 15: 537-541). At least one method for increasing pear fruit shelf life fragrance for ODNs library and (Ho and so on, (1996) Nucleic Acids Res. , 24: 1901-1907; Birikh and so on, (1997) RNA 3: 429-437; with Lima and so on, (1997) J.Biol. Chem., 272 =626-638) „and monitored getatable position (card further Birikh VACCINE through zyme H cutting and so on, with; With Ho and so on, (1998) And Biotechnol. , 16: 59-63) „The RNA zyme H of catalyze DNA-RNA transfusion DNA RNA one hydrolyzing cutting of di-phosphate and principal chain.

[0221] The and method for increasing pear fruit shelf life friction, relates to the use of the other stochastically, a of the body ODNs storehouse, which is of an appraising VACCINE of zyme H cutting in vitro synthetic RNA improves the position of the getatable. Toilet paint is extended and evaluating target molecule of said position (card further Lima and so on, with). For VACCINE is the other method for increasing pear fruit shelf life capable of trans- target computer is one-third to RNA foldable model. A report already claimed is made ribozyme and library screening effective cutting (card further Campbell and so on, (1995) RNA 1: 598-609; Lieber and so on, (1995)Mol. Battery Biol. , 15: 540-551; with Vaish and so on, (1997)Biochem. , 36: 6459-6501).

[0222] Is ODNs VACCINE and zyme and jointed or the other in vitro method for increasing pear fruit shelf life capable of the library and computer analogue is possibly more effective (Lima and so on, with). Natural, in vitro synthetic use of VACCINE prediction cannot trans- ODNs of the accessibility of body, the one diameter of the annealing interactions of polynucleotide (card further of the effect of RNA- binding protein recently Tsuchihashi and so on, (1993) Science, 267: 99-102; Portman and so on, (1994) EMBO. J, 13: 213-221; With Bertrand and Rossi (1994) EMBO. J, 13: 2904-2912). The content of punished the simulating patent number of the article used to 6,562,570, provided is a dual-head battery extractive condition of confirm mRNA of the adjacent position of compositions and methods for increasing pear fruit shelf life friction, wherein the extractive with simulated the body condition.

[0223] Simple net that the method for increasing pear fruit shelf life fragrance methods of the hybrid conditions of the reaction culture medium, the culture medium comprising an endogene RNA- binding protein's battery extractive, or one or multiple types of RNA- binding main protein the battery extractive and a result of the dual-head, such of the natural or synthetic form RNAs and particular trans- ODNs, ribozyme or DNAzymes, or and or an and 0DN, ribozyme or DNAzyme library with A Temperature. With) RNA end of a position complementary any trans- 0DN, ribozyme or DNAzyme, which can with a of hybrid. The use particular's ODNs or the ODN library, the RNA zyme H the dual-head during the hybrid, the capable of the other side of the hybrid, scissors already of hybrid VACCINE. The use ribozyme or DNAzymes, can vertically RNA zyme H, and varies, essential, wherein with the hybrid wherein the ribozyme and DNAzymes can automatically VACCINE. A plurality situations, comprising for the battery and extractive or an ODN and library, wherein a endogene mRNA and RNA- binding protein and RNA zyme H.

[0224] Toilet, wherein with a of method for increasing pear fruit shelf life fragrance evaluating) RNA and already unified trans- ODNs, ribozyme or DNAzymes, and provided with a cutting position of. For example, wherein relying (card further for the aim of the well transferase polymerase chain side (TDPCR) Komura and Riggs (1998) Nucleic Acids. Res, 26: 1807-11). Records the steps with replaceable the RNA template through the reversal DNA, comprising perform TDPCR. The invention, TDPCR method for increasing pear fruit shelf life fragrance required 3 ') fixes a proper dependent of the RNA DNA polymerase (e.g.. the transcriptase) records the generating to the one) RNA perform reversal. The aim through letting a ODN paint (Pl) with RNA) RNA molecule part other area (S located at a mounted 卩, along the RNA molecule is 5 to -3 of direction) hybrid. The present invention dNIPs, wherein polymerase from Pl 3 ') of DNA the copying VACCINE, and is trans- 0DN/RNA zyme H, ribozyme or DNAzyme generating cutting position end copying. The novel DNA molecule (is a dart first one DNA) is fixed TDPCR method for increasing pear fruit shelf life fragrance PCR part and a role of the first template, which is of an appraising the current is RNA corresponding getatable the target sequence of.

[0225] For example, comprising capable of the TDPCR method for increasing pear fruit shelf life friction, namely, and reversal of guanosine three phosphoric acid (rGTP) record DNA is arranged on the reaction of the tail are well transferase (TdT) effect, comprising a DNA molecule 3 ') further rG (2-4) tail. And the dual-chain ODN joint, comprising with 3 ' extensions any one, and rG (2-4) tail base group pairs. And adding two PCR primers. Head is connected paint (LP), with TDPCR joint one complementary, wherein a on the rG) tail holes (sometimes is dart and lower one). The paint (P2) is possibly same as Pl, and can be with respect to Pl sleeve, namely, which is located at least one part with) RNA of the Pl and area upstream's area complementary (i.e.; the RNA molecule 3 to -5 of direction), wherein, comprising VACCINE target molecule other part is located at a research. Frame is, a ventilate research of the confirm wherein a getatable connecting site's) RNA molecule's part, wherein the complementary the P2 part of area upstream's. And the current DNA polymerase and dNIPs state with the known perform PCR, to increase of the DNA fragments of paint LP and P2 the limit. Toilet of by multiple types of known method for increasing pear fruit shelf life capable of any an internal constricting the product of increasing, and afterward with automatic DNA sequencer sequence, for providing to cutting position of dried precisely. At the confirm the property, synthetic can confirm sequence trans- DNA or the ribozyme, comprising in vitro with the body of.

[0226] The trans- intervention the outer particular gene is connected with and synthesizing trans- widowed nucleotide sequence card (, for example, Lefebvre-d' Hellencourt and so on, (1995) Eur. Cyokine. Netw, 6: , 7Agrawal (1996) TIBTECH, 14: , 376Harvester grain opening Lev-Lehman and so on, (1997) Antisense Therap. Cohen and Smicek, eds. (Plenum Filtering, Novel York)). Simply speaking, the trans- widowed invention can sequence of the short DNA sequence, usually is 15-30 poly body; further, capable of the card further Wagner to 7 poly body and so on, (1994) And 372: 333), Which is designed with one target mRNA complementary, and a VACCINE the =AS transfusion DNA. A layer of such transfusion DNA for preventing the processing, film editing, conveying or a linearly related mRNA. Moreover, wherein multiple AS nucleotide sequences with their target mRNA hybrid of with battery zyme VACCINE H movable, thus of the mRNA degradation (card further Calabretta and so on, (1996) Semin. Oncol. , 23: 78). The such sleeved, the RNA zyme H is fixed to the transfusion DNA's RNA ingredients, and can latent release AS, comprising further with the molecular hybridizations of target VACCINE. A AS with genomic DNA shape generating the other one affects the vibration, wherein the shape of a tertiary the spiral, comprising a copying of the.

[0227] And the non- of one of trans- sequence claims a text discussed that supplemental or vicarious example, be used for the ribozyme the suppressor gene function. When the trans- combined therapy of the factor ends of chemometry, the is in essential. And capable of the target location groove in sequence the ribozyme. The ribozyme is connected with the RNA catalysis ability RNA molecule, which can automatically the special locating part is high VACCINE. Is greater than of ribozyme cutting the number of RNA molecule of 1: The number of 1 chemometry (extrapolated card further Hampel and Tritz (1989). Biochem, 28: 4929-33; Harvester grain opening Uhlenbeck (1987), And 328: 596-600). Therefore, the invention is capable of the ribozyme sequence, wherein the target sequence to fixed PDGF or the VEGF mRNA type getatable field, and a proper taking centre. According to the field of male knowledge and article further a method for increasing pear fruit shelf life capable of discussion preparation and delivery ribozyme. The ribozyme of trans- sequence combined use.

[0228] The ribozyme of catalyze di-phosphate and a cutting of VACCINE. With already appraised a plurality of ribozyme structure families, comprising I and intron and RNA zyme P and hepatitis D virus ribozyme, the hammer head ribozyme and a hairpin ribozyme, which is from the tobacco ring part and satellite RNA (sTRSV) minus including (card further Sullivan the first (1994nnvestig. Dermatolog (Suppl. 103): 95S; With simulating patent of 5,225,347). The front two and families on the virus of viroid; the ribozyme is considered capable of separating unit and joint widowed poly body during the sliding ring (card further Symons (1989) TIBS, 14: 445-50; Symons (1992) Ann. Rev. Biochem. , 61: 641-71). The hammer head and a hairpin nuclear substrat foreword commonly the is of a a gene therapy anti-form cutting of mRNAs. The invention according to the field known method for increasing pear fruit shelf life the fragrance for ribozyme type. A hairpin ribozyme is used for clinical trial now, and is in for type. generally, the duration of ribozyme is 30-100 nucleotides.

[0229] Configured for catalyzing the ribozyme molecule of cutting target mRNA duplication for the field the male knowledge (e.g.. PDGF (SEQ ID NO: 1) Or VEGF (SEQ ID NO: 3), And further capable of preventing mRNA (translator card, for example, PCT international of claims WO; 90/11364Sarver and so on, (1990) Science, 247: 1222-1225 With simulating patent of 5,093, M6). Although of reign specific identification sequence scissors mRNA ribozyme capable of prodrugs of mRNAs particular, and use of hammer head ribozyme is specifically for. The hammer head ribozyme can automatically mRNAs of the flanking one area, wherein flanking area is provided with a complementary the base group of a target mRNA. The pedal requirement, target mRNA at the following sequence of base in: `5) Ug-3 `. The structure and production of hammer head ribozyme of the field the male knowledge, and are described of the following literature =Haseloff and Gerlach ((1988), And 334: 585).

[0230] Ribozyme farther comprising VACCINE contact is ribonuclease the invention (lower of the Cech- ribozyme”), to) and present's the zyme the thermophile tetrahymena CTetrahymena (thermophila is dart and IVS, or VACCINE L-19IVS), and a already and colleague perform of Thomas Cech the introduction the card (further of the following literature Zaug and so on, (1984) Science, 224: 574-578; Zaug and Cech (1986)Science,231: 470-475; Zaug; said (1986), and 324: 429-433; An international monopoly application of W088/04300; Washbasin and Cech (1986) Battery, 47: 207-216). The Cech- ribozyme with eight bottom pairs' movable position part, which can with a target sequence VACCINE hybrid, comprising a cutting of target VACCINE. The invention layer comprising the Cech- ribozyme, wherein a target eight pairs of a flexible position sequence. Although the invention not limited to operating mechanism particular theory, and hammer head ribozyme the invention use, for PDGF/VEGF- direction detection's trans- compared with the advantages; because of the newest alarming centigrade, the hammer head ribozyme is blocked VACCINE the control and/or from cutting of mRNA target with an effect.

[0231] Heat-insulated to the trans- method for increasing pear fruit shelf life stick, so that the ribozyme capable of the oligonucleotide comprises for modified nick (e.g., stability for improving and targeted part), and a section of the is outer target mRNA the pool. For delivering method for increasing pear fruit shelf life capable of claims a port to the ribozyme the DNA structure body, comprising a strong and pol III or the control of pol the promoter, messenger and inhibiting for movement for the battery infections by administration capable of generating enough are ribozyme, fixed by the clamping target. At the ribozyme and trans- molecule of different, wherein the catalysis, so the utility the lower battery the concentration to realize wherein potency.

[0232] Such as claims text the; the required the words, nuclease anti-skid through the field of any method for increasing pear fruit shelf life capable of providing male knowledge, which is not disturb the trans- oligodeoxynucleotide or the ribozyme are arranged on the whole, common to for and delivery method for increasing pear fruit shelf life fragrance required (Iyer and so on, (1990) J. Org. Chem. , 55: 4693-99; Eckstein (1985) Ann. Rev. Biochem., 54: 367-402; Spitzer and Eckstein (1988) Nucleic Acids. Res, 18: 11691-704; Woolf and so on, (1990) Nucleic Acids. Res, 18: 1763-69; with Shaw and so on, (1991)Nucleic Acids Res. , 18: 11691-704). Heat-insulated to claims text, a to the trans- oligonucleotide or ribozyme perform according non- one modified representative for described body, in nuclease anti-skid, the modified phosphate principal chain the back containing or the oxygen hetero-atoms, short one alkyls or the naphthenic bottom saccharides the button or the short one hetero-atom or the heterocycle saccharides the key. Which comprises, for example preparing 2 ' - fluorizating, 0) methylation, phosphine methyl acid esters, sulfo- phosphates, dithio- phosphate and morpholine generation of widowed poly body. For example, wherein trans- oligonucleotide or ribozyme of the sulfo- phosphate key, wherein the four of six 3 '- the invention base groups. Moreover, the sulfo- phosphate key is connected with the invention is made of. Sulfo- phosphate trans- oligonucleotide of effective, and animal body manifestation and enough pharmacokinetics half-life period of the concentration usually the manifestation hole (and a obvious toxicity (card further Agarwal and so on, (1996) TIBTECH, 14: 376), And the nuclease anti-skid. Moreover, a AS-ODN nuclease anti-skid of the through 3' )is formed with nine invention providings sequence, wherein the invention is sequence CGCGAAGCG. Avidin the use of idrabiotaparinux binding reaction, further capable of improving AS-ODNs protection action the blood serum nuclease degradation (card further Boado and Pardridge (1992) Bioconj. Chem., 3: 519-23). According to concept such, wherein every 3 ' - end of AS-ODN reagent perform sole idrabiotaparinux. With the avidin with reaction time, they are formed the energy-saving, nuclease anti-skid compound, and is preferably binding ODNs, and 6 times of higher stabilities.

[0233] The research already with trans- oligodeoxynucleotide extending body (Agarwal and so on, (1991) Proc. Natl. Acad. Sci. USA (88): 75%). According to the external, the method for increasing pear fruit shelf life capable of being used as is a exception the eliminating mechanism of AS- oligonucleotide from the circulating stone arranged; the is fixed by and 3 ' )is ladder-shaped is provided with a oligonucleotide of fixing drifted with. Therefore, wherein the upper sulfo- phosphates, the ring protections on or idrabiotaparinux key position of avidin ensure surface of as- oligodeoxynucleotide foaming the stability.

[0234] Connecting base group of using the modified piece, and preparing the invention analogues; the structure of nucleotide with a base change, wherein the invention analogues is as the therapeutic agent or the experiment reagent. The example of the invention is analogues peptide nucleic acid (PNA), DNA (VACCINE or) ribodesose (or ribose) phosphate principal chain substituted for of the polyamide principal chain, comprising with a production of the invention is of said peptide with. Already the display PNA analogues of the degradation function of antienzyme, and body in vitro with extended life. Moreover, already display PNAs and cDNA sequence is strong than the DNA molecules. The so that the is at one PNA and DNA one deficient disposed between the rechargeable repel. A card (further comprises a polymer principal chain and morpholine generation of polymer chains principal to the modified of oligonucleotide perform, for example, simulating patent of 5,034,506, wherein the literature content is punished the article reference), the principal annular chain or the ringless principal chain, saccharides use or any modified, comprising a improving oligonucleotide pharmacokinetics the modified.

[0235] The invention; the other hand claims for DNA for zyme target mRNA lighting, for example, PDGF or VEGF. The DNA zyme with conformed with the trans- and multiple machine-made characteristics of ribozyme technology. Below the DNA zyme, thus they are respectively a particular the target-type nucleic acid sequence, the photo is a trans- oligonucleotide, wherein, they can catalyze and specific targeting cutting nucleic acid thereof is a ribozyme.

[0236] At are two types fundamental DNA zymes, wherein two types of zymes is formed Santoro and Joyce evaluating (card, for example, simulating patent of 6,110,462). the 10-23DNA zyme comprising a ring structure, the structure is two arms. Are two arms are identification particular target nucleic acid sequence provided with a specific, and annular structure providing the taking function of physiological condition.

[0237] Simple net that part of the specific the identification and cutting target nucleic acid DNA zyme, the technical field for firsts surface appraise the cloth the target sequence. The capable and the trans- oligonucleotide same method for increasing pear fruit shelf life fragrance realizes. The multiple examples, wherein cloth or a sequence substances of the is rich in G/C and 18-22 nucleotides. And G/C contents of conveying to ensure the DNA zyme and strong shape between the target sequences.

[0238] When the synthetic DNA zyme, capable of the collection trans- identification sequence of zyme targeted messenger is, thus wherein comprises DNA zyme two arms, and placement of the DNA zyme's ring are two arms special.

[0239] Preparation and employs the DNA zyme method for increasing pear fruit shelf life fragrance card, for example, U.S 6110462. . , In vitro of the or form of DNA the ribozyme the method for increasing pear fruit shelf life fragrance comprising to deliver RNA ribozyme of the article elaborated the inverse method for increasing pear fruit shelf life stick. Moreover, the technical field personnel is connected with is similar to the trans-, oligonucleotide of DNA to zyme perform optionally modified, comprising a improving stability and improving anti-skid to degradation.

[0240] RNAi antagonist

(0241) By multiple plans the invention for is provided disturbed (RNAi) to realize VEGF and PDGF inhibit and a method for increasing pear fruit shelf life through friction VACCINE. The RNAi is a sequence from duplication, a gene and method for increasing pear fruit shelf life friction, which can occur of the eukaryotic battery. Generally, the method for increasing pear fruit shelf life fragrance claims connected with the homology sequence the dual-chain RNA (dsRNA) capable particular sequence mRNA degradation. For example, is not particular to a stranded mRNA (SS mRNA sequence) is dsRNA outer; the motor is connected with the messenger and is unstable, thus in corresponding a gene outer. Therefore, by inducing dsRNA for inhibiting any gene of aid, wherein dsRNA is equal to the gene mRNA completely or the main part. A seems to, wherein the outer dsRNA, the first producing at the duration to a through the ribonuclease III to 21-22 pairs of short dsRNA oligonucleotides. Therefore, RNAi can be realized by with respect to short homology dsRNAs inducing or outer. The fact, use of respect to short homology dsRNAs, possibly with multiple advantages, common to the following discussed.

[0242] The mammalian cell with wherein dual-chain RNA (dsRNA) is at least two paths. The RNAi (sequence from) is path, the first front dsRNA the decompose Are in Segment (si) RNAs, such as claims text the. siRNAs with a 21 nucleotides with righteousness and trans- sliding, comprising a respective 3 ) is a left 19 nucleotides with two nucleotides a extensions si RNAs. Short in RNAs is considered is provided with allowed the target at the particular messenger RNA perform degradation the information sequence. On the insertion non-, specifically path connected with dsRNA trigger of any sequence, a long and duration wherein at least is about 30 of pairs of line. The utility model of nonspecific motion of the dsRNA (activates protein kinase of two types of zyme JKR dual-chain RNA- active), wherein the form to the control initiation factor eIF2 perform phosphorylation, comprising sealing of protein synthetics, and 2 to 5, widowed a adenylate synthase O', 5 ' - aq, wherein synthetic's molecule capable of activate RNA zyme AN, namely one target-type thereof is mRNAs non- collection zymes. The non- specific with possibly to represent the main or virus infection to the response tank, generally, wherein the effect of non- specific with the invention to obtain the minimum of the specifically for method for increasing pear fruit shelf life stick. The deserve to bushing is, and multiple steps long dsRNAs to redirect teat dip to the sensor non- specific path, therefore, short is realizing gene and is RNAi the left 30 is pairs' dsRNAs is specifically for (card, for example, Hunter and so on, (1975) J. Biol. Chem., 250: 409-17; Manche and so on, (1992)Mol. Battery Biol. , 12: 5239-48; Minks and so on, (1979) J. Biol. Chem., 254: 10180-3; With Elbashir and so on, (2001), And 411: 494-8).

[0243] For realizing RNAi the durations of multiple dual-chain oligonucleotides is less than 30 of pairs; and possibly comprising a ribonucleic acid about 25, 24, 23, 22, 21, 20, 19, 18 or 17 is pairs. DsRNA oligonucleotide the invention optionally comprising 3 ' telescopic. The non- demonstration's one of nucleotide 2) 3 ' telescopic capable of any type ribonucleotide elastic members, and are capable of `2 )- triterpenes by the elastic members; and the cost VACCINE of synthetic, and improving siRNAs, the battery culture medium and nuclease anti-skid of the battery piece of the transfection (card further Elkishi and so on, (2001), And 411: 494-8).

[0244] With 50, 75 and 100 or 500 of bottom or pairs of multiple long dsRNAs further capable of multiple by plans the invention. For integrating the RNAi dsRNAs rotary concentration is about 0. 05 ηΜ, 0. 1 ηΜ, 0. 5nM, 1.0 ηΜ, 1.5 ηΜ, 25 or ηΜ ΙΟΟηΜ; and factors of the property and a gene of targets and technical field for although nick according to the treatment of conveniently know capable of the concentrations. Demonstration's dsRNAs of the chemical body or the use are type vector production in vitro the or form. Demonstration's synthetic RNAs comprising of the field known method for increasing pear fruit shelf life capable of synthesis 21 nucleotides RNAs (e.g.. accelerates the phosphoramidite VACCINE and chest phosphoramidite triterpenes by (ftOligo, Germany)). Synthetic oligonucleotide capable of the field male knowledge the method for increasing pear fruit shelf life fragrance to escape the protection, and gel tail (card further for example, Elbashir and so on, (2001) Genes. Dev, 15: 188-200). Preferably long RNAs capable of promoting the duplication, so the field male knowledge T7RNA polymerase promoter. A copying to the target's two chains in vitro of the promoting sole downstream's RNA) along two types of the direction, is formed with required target sequence dsRNA oligonucleotide.

[0245] For a particular sequence of oligonucleotide capable of the comprise a lighting any continuous nucleotide sequence of the target gene messenger (e.g.. PDGF sequence (e.g.. SEQ ID NO: 2) Or VEGF sequence (e.g.. SEQ ID NO: 4). Capable of the field known procedure and algorithm the proper target sequence. Moreover, wherein and the best sequence according to claims text, for prediction to particular single stranded nucleic acid sequence secondary structure, and space are can be socket the foldable mRNA exposing device for which the sequences of single stranded area for the design of. Oligonucleotide the method for increasing pear fruit shelf life fragrance and composition for is formed a card, for example, the simulating patent of 6,251,588, wherein the literature the content is punished the article finger pricking device. Generally, mRNA is considered of the linear molecule, comprising a command protein synthetic information the invention ribose sequence. Natural, the research already promulgates on the second-level block and tertiary structure of majority of mRNAs. The secondary core structure of majority VACCINE of the same RNA molecule watson between the zones of salts of the interaction layer of Krick type. Important secondary structure and a molecule is compounded with area, hairpin ring and a bump and inner ring of the transfusion DNA VACCINE. When the tertiary structure element of the secondary structure and each other unit or a single stranded contact area formed, comprising a three-dimensional structure of generating compound. A lot of researchers with already surveyed the block RNA transfusion DNA structure energy-gather, with inferred capable to prediction of the secondary VACCINE structure a switch of rules (card further for example, Jaeger and so on, (1989) Proc. Natl. Acad. Sci. USA (86): 7706 (1989); A Turner and so on, (1988) Ann. Rev. Biophys. Biophys. Chem., 17: 167). The conveying device capable to appraise the VACCINE element structure, and wherein said that a appraising the single stranded RNA area, comprising possibly represented mRNA targeted with silenced RNAi, ribozyme or the anti-sense technology of the connected with. Therefore, wherein appraise the mRNA target's particular connected, is used for part to drive the RNAi dsRNA oligonucleotide, and is formed ribozyme and hammer head ribozyme composition the invention.

[0246] A fixes heterologous target gene transfection the dsRNA oligonucleotide inducing battery, and field male knowledge such as liposome's carrier composition, for example (Lipofectamine2000 Technique Life, Rockville MD), common to the tube is describes around the adherent cells. The target-type of the endogene gene the dsRNA oligonucleotide transfection is made Oligofectamine (Life Technologies) perform. Overturning mammalian cell line; and the transfection codes hGFP pAD3, a working for (KehlerAack through the fluorescence microscope for examination transfection and so on, (1998). J Battery. Biol., 141: 863-74). The RNAi potency, the inducing dsRNAs adopts in multiple types of measuring method for increasing pear fruit shelf life fragrance afterward a appraises circuit. The method for increasing pear fruit shelf life incense comprises, and down in limiting to of protein and signature of antibodies, wherein antibody of the novel protein synthetic inhibit such endogene storehouse turning enough time afterward respectively gene product according the target-type fixed that and VACCINE and signature, for determining the existing target mRNA layer.

[0247] And compositions for RNAi technology, method for increasing pear fruit shelf life and capable of providing the invention is the following literature: Simulating patent of 6,278,039,5, 723,750 or 5,244,805, wherein the literature is punished the article finger pricking device.

[0248] Receptor tyrosine kinase inhibitors antagonist

[0249] The invention farther comprising field the male knowledge's tyrosine activating enzyme antagonists and raised form and substitute, wherein a raised and substitutes capable of folium artemisiae technology the field to obtain, and punishes the article to periphery prior art teaching. PDGF (and VEGF the extracellular signal through a of and the other part of micro-processing of phosphorylation event and battery according of tyrosine activating enzyme to the PDGF receptor (and VEGF receptor) realized; the effect of battery and a protein of signal conductive compound other. Therefore, affects the PDGF (and/or VEGF) and conductive antagonists further capable of embodiment method for increasing pear fruit shelf life capable of acceptor activating enzyme in the invention.

[0250] Or the VEGFR tyrosine activating enzyme receptor zyme with big multiple types of type's the tyrosine kinase inhibitors are such as PDGFR is known (card, for example, Spada and Myers ((1995) Exp. Opin. Ther. Patents,

5: 805) And Bridges ((1995) Exp. Opin. Ther. Patents,5: 1245). Moreover, Method and Lydon with already cantilever the tyrosine kinase inhibitors anti-tumour body ((1996) Emerging Medicine =The Prospect Is Improved Medicine, 241-260). For example, the simulating patent are described 6,528,526 of substituted 喹 oh 啉 the compound, which can selectively platelet-derived inhibiting growth factor receptor and (PDGFR) tyrosine activating enzymatic activity. PDGFR tyrosine activating enzymatic activity known inhibitor comprising quinoline is subjected, comprising Maguire, (1994) J. Med. Chem. , 37: 2129) And Dolle, said that (1994) J.Med. Chem., 37: 2627) Alarming. An type based on phenyl amino and pyrimidine coating recently of Traxler and so on, wherein EP 564409, comprising Zimmerman and so on, (1996) Biorg. Med. Chem. Lett.

6: 1221-1226), And a Buchdunger and so on, (1995) Proc. Nat. Acad. Sci. (USA),92: 2558) Alarming. Capable of inhibit PDGF receptor tyrosine kinase movable quinazoline derivative comprising of and the card (further with the bicyclo aryl compound and mixing aryl compound, for example, WO 92/20642), 喹 oh 啉 the flat plate (card further (1994) Res Cancer., M: 6106-6114), Pyrimidine derivatives member (patented claims a claim. 87834/94) Is of a root quinoline derivative (card further Abstracts of the 116th Annual Sections of the Medicinal Society of Japan (Kanazawa) (1996), 2, p.275, 29 (C2) 15-2).

[0251] The example of VEGFR tyrosine kinase inhibitors comprising a cinnoline derivatives, for example, claims a simulating patent of 6,514,971, wherein the content of the patent punished the article to periphery of every foot text a. Such cinnoline derivatives similarly known, for example, (1995) J. Med. Chem, 38: 3482-7 Claimed is 4) (3) monobromo-benzene amidogen); cinnoline(1968) J. Chem. Soc. , C (9): 1152-5 Claimed is 6) and one 4) phenoxy; cinnoline(1984) J. Karnatak Univ. , Sci. , 29: 82-6 Claimed of multiple 4) anilino-; cinnolinesWith 1973nndian (J. Chem. , 11: 211-13 Claimed of multiple 4) phenyl sulfo- cinnolines. Moreover, (1973) J. Karnatak. Univ, 18: 25-30 Claimed of multiple 4) phenoxy cinnolines, (1984) J. Karnatak. Univ, Sci. , 29: 82_6 Claimed two kinds of compound: 4) (4) Methoxy anilino- -6,7) of each oxygen root and cinnoline 4) (3) chlorobenzene amidogen -6,7) of each oxygen support cinnoline. Moreover, wherein 4) are located by selected from O-, S-, NH- and CH2- group and multiple cinnoline descriptions of the benzene ring of the following literature: Simulating patent of 5,017,579, simulating patent of 4,957,925, simulating patent which 4,994,474 and EP 0302793A2.

[0252] And/or PDGFR and related compound for inhibiting VEGFR to obtain the target-type receptor tyrosine kinase movable effect through the screening novel compound, the variable for determining the method for increasing pear fruit shelf life stick. Candidate PDGFR or the effective inhibitory effects of VEGFR micromolecular organic coating, capable of battery and to the measuring system monitor, and this field the other measuring system of male knowledge.

[0253] For example cloth, aiming at the VEGF- receptor tyrosine kinase's movable detection an is as follows. The detection to the Flt-IVEGF- receptor tyrosine kinase perform. The detailed and is as follows: The room temperature 20mM solution Tris. HCl pH 7. 5, 2-3 manganous chloride (MnCl2), 3mM magnesium chloride (MgCl2) and 10 μ END of vanadate, 0. 25mg/ml polyethylene glycol socket (20000), Is dithiothreitols and 3 μ g/.mu. 1 Poly (Glu and Tyr 4): 1 (Sigma, Buchs, Switzerland), 8 1 [3] and said Fixing Each (0.2 )is), 1% dimethylsulfoxide harvester grain opening 0-100 μ END clearly the detection 30 μ 1 activating enzyme solutions the compound (IOng the Flt-I activating enzyme for (card further Shibuya; said (1990) Oncogene, 5: 519-24) Is matched and breeding temperature is 10 minutes. A through adding 10 μ 1 to 0. 25 Μ edetic acid (EDTA) pH 7 terminations of the reaction. Use multi-channel for (LAB SYSTEMS, the simulating), a 20 μ 1 aliquots (PVDF = poly two vinyl fluoride (polyvinyl difluoride)) Immobilon P film (Millipore, simulating), a micro titrate filter branch tube, and vacuum connected. The full-automatic removing liquid, washing and film of a 0.5% phosphoric acid (H3PO4) baths first four times, and washing with a ethanol time, the vibration each breeding a temperature is 10 minutes, comprising mounted on Hewlett Packard TopCount Pipe, and adding 10 μ 1 Microscint. RTM. (β and scintillation counter liquid) afterward measuring the radioactivity. Are three or more of concentrations each compound the linear regression assay of IC5tl- value of inhibiting percentage (concentration usually is 0. 01 μ mol, 0. 1 μ mol and μ 1) mol. The IC5tl- value of tyrosine movable covering compound can be at the μ 0.01 M-100 μ END ranges.

[0254] Moreover, VEGF- induce's VEGFR tyrosine activating enzyme/of phosphorylation movable a test with the other verifies on the battery. Simply speaking, the permanence of the outer CHO battery inoculate for of VEGF receptor (VEGFR/KDR) transfection piece of (a fetal 10% serum calf 05) to the maximal media of said battery culture) are, And is 371: Upper, The Temperature of 5 %0 (2), wherein left 80% convergence from the manifestation. (Does not contain FCS the culture medium, a 0.1% ox blood cleaning proteins) dilutes clearly detecting the compound, and to further the battery (comparison comprising does not contain experiment compound culture medium). 37°C Breeding a temperature is 2 in method, adding ornamental VEGF, the tail VEGF concentration is 20ng/ml). 37°C A breeding heating temperature is 5 minutes, in ice-cold PBS a battery twice; and decomposes each 100 μ 1 decomposition the turning infusion device. Toilet to decomposition liquid perform centrifugal, removing nucleus battery, and use commercial protein determining (BIORAD) to determine the liquid supernatant protein concentration. The decomposition liquid afterward directly to use, or is required of the 200°C a storage.

[0255] Toilet perform a filling ELISA, a measuring KDR- receptor phosphorylation: The KDR monoclonal antibody fixed on the black ELISA plate (buys from I^ckard OptiPlate™, HTRF-96). A washing to the plate, and a solution PBS other BSA saturated free protein binding sites. And the battery decomposition liquid OOyg protein/hole) and alkaline phosphatase coupling's resistance phosphotyrosine antibody (e.g.. PY20: AP buys from Transduction Laboratories, Lexington, KY) of the 4°C, wherein the Temperature plate Is connected with the luminous. Washing to the plate at each, comprising for generating to 7 )AP/)and (CDP-Star piece, to use, with Emerald; theApplied-Biosystems TROPIX Bedford, MA) verifies resistance phosphotyrosine antibody and with the connecting of phosphorylation receptor. Measuring the lighting with I^ackard Upper With Microplate Scintillation Counter. Is comparing (with VEGF or the PDGF collaterals) and negative comparison (not of VEGF or the PDGF collaterals the extending, signals are equal to the VEGF- capable KDR- receptor phosphorylation = (100%). Test material layer and VEGF- capable KDR- receptor phosphorylation % inhibiting computation, sensor, maximum inhibitory effects 1/2 of the material the concentration is defined is ED5tl (effective doses of inhibitory 50%) actions. The ED50 value of tyrosine movable covering compound of the 0. in 001 μ Μ -6 μ ranges END, usually is 0. 005 μ -0 Μ 5 To μ END and.

[0256] And drug and combined therapy employ

[0257] The Resistance of vegf and Resistance of pdgf agent capable of combined therapy novel vascular diseases, comprising psoriasis, rheumatoid arthritis and eye novel vascular diseases. Electric specifically is interested is for PDGF-B antagonists compound and VEGF-A combined antagonist inhibit such as the yellow part denaturation or diabetes mellitus retinopathy the method of treatment of eye novel vascular diseases. Therefore, at diagnosis patient with emergency to developing or flexible such disease of the novel vascular diseases, by administering an PDGF antagonists and antagonist VEGF combined to the patient perform combined therapy, which is a effect of PDGF and VEGF, thus development for inhibiting novel vascular diseases, and relieved and vascularization related adverse effect. The invention method for increasing pear fruit shelf life fragrance embodiment, is not adjust the cornea dropsy. Heat-insulated to claims text discussed that capable of a multiple types of PDGF and VEGF antagonists the invention.

[0258] According to the invention Resistance of pdgf and Resistance of vegf combined combined therapy of the independent perform or other and combined therapy combined performs, and capable of the home, medical offices, clinic and hospital outpatient clinic or the hospital providing. Generally, the combined therapy opened on the hospital, so the doctor is tightly observe the treatment effect, and perform any required seasoning. The duration of combined combined therapy is fixed by wanting combined therapy's novel vascular diseases the bumps and conditions of and disease patient type and patient end and type and a response to the combined therapy. Moreover, comprising a development for the novel vascular diseases large risk of (e.g., diabetes) of the holding combined therapy, comprising a inhibiting or the outbreak of delaying to sign. The invention provides obvious advantages, is used for combination therapy novel vascular diseases the PDGF antagonists and VEGF antagonists combined, such that can employ the lower dose each antagonist, and at general movable antagonist, accordingly of low toxicity and side effect and low-cost providing production effect.

[0259] Combined combined therapy the alleviation of each of the antagonists of through a proper method for increasing pear fruit shelf life fragrance perform, comprising a sleeve with the antagonists sections of the antagonists the concentration is a effective combined therapy novel vascular diseases. For example, wherein a antagonists of a proper carrier material mixing, and generally wherein buffer for accounts is a composition total weight 1_95%. The composition is further can close to the eyes suitably, an opening and stomach (e.g., wherein the vein, skin, hypodermic), an, inhaled the alleviation of the peel and Nose Huo the dosage form providing. Therefore, wherein the composition is the following form, for example, the tablet, capsule, pill, powder, particles, oat suspension, latex, and a gel (comprising hydrogel), paste, paste, creams and ointment, deliver device, suppositories and enema injectable, implant, spray or aerosol the. A antagonists one or two or more or more than two kinds antagonists of the pharmaceutical composition according to the common method and method for increasing pear fruit shelf life fragrance preparation (card, for example, Remington: The Science and Practice of Pharmacy, 20th ed. ) Ed. A R. Gennaro, 2000, Lippincott Williams & Wilkins, Philadelphia, PA. harvester grain opening Encyclopedia of Pharmaceutical Technology, Eds. , J. Swarbrick harvester grain opening. J C. Boylan, 1988-2002, Marcel Dekker, Novel York).

[0260] The internal surface, PDGF and antagonist VEGF combined perform stomach impressed heel (e.g.. or implant or the body-wide are connected in a of the skin, peritoneum, vein, intraocular, vitreous body, retrobulbar, conjunctiva, subtenon or subcutaneous injection). Outer or drug are body-wide for stomach or non-aqueous comprising a sterile aqueous solution, oat suspension or emulsion. Capable of multiple types of water carriers, water, in water for example, the water and buffering cross and equal. The examples of the formed intermedia comprising poly propylene glycol, polyethylene glycol, plant oil, gelatine, hydrogel and hydrogenation naphalenes and injectable organic ester, 0.5-2 to oleate. The drug further comprises a assisted material, to the agent, wetting agent, buffering agent, emulsifying agent and can agent. Biodegradable compatible, lactide biodegradable polymer and lactide/glycollide medical or polyoxyethylene is poly oxygen comprising and capable of the control release of active ingredients.

[0261] Moreover, capable of to employ PDGF and antagonist VEGF combined by the opening. For oral taking the composition for in preparation of solid or the liquid form, comprising according to any known method for increasing pear fruit shelf life fragrance perform of pharmaceutical composition realm for producing.

[0262] For oral taking solid dosage form of a are capsule, tablet, pill, powder and granule. Generally, wherein the drug comprises a sleeving of the agent excipient mixing active components (for example PDGF small molecular organic antagonists and VEGF small molecular organic antagonists). The excipient comprises, for example, inert diluent, calcium to carbonate, sodium carbonate, lactose, cane sugar, glucose, mannitol, cellulose, starch, calcium phosphates, sodium phosphates, kaolin. Further capable of the adhesive, buffering agent and lubricating agent (e.g., sodium starch) glycolate. The tablet and pill further capable of the enteric clothes is prepared. The composition can be optionally comprising increasing the stevia agent, flavoring agent, colouring agent, aromatizing agent and sterilizing agent, comprising a providing the delicious drug.

[0263] For example, wherein PDGF and VEGF antagonists capable of the injection perform are intraocular through the vitreous body; the opening, and a perform conjunctiva and subtenon injection. The paths is a back part of the scierotic, retrobulbar, peritoneum, and skin according. Moreover, antagonist combined capable of drug to deliver device or a intraocular implant section (card is as follows).

[0264] For oral taking liquid dosage form of a are capable and emulsion, solution, oat suspension, syrup and soft capsule. Are formed comprising the field of commonly inert diluent, to water or oil medium, and can also comprises a adjuvant, to wetting agent, emulsifying agent and a.

[0265] A plurality situations, PDGF and VEGF antagonist is combined with a topical application, for example, and patch or through a are directly of the area, such as the trouble or flexible the novel vascular diseases intraepithelial or the sleeve, or adopts the iontophoresis.

[0266] For eye and use and capable and excipient mixing drug comprising a active components' tablet. The excipient is, for example, inert diluent or filler (e.g., cane sugar and sorbitol), lubricating agent, glidant and resisting cement (e.g., sodium starch glycolate, zinc stearate, stearic acid, silicon dioxide, hydrogenated vegetable oil or speckstone).

[0267] The PDGF and VEGF antagonists or intermedia and other tablets, or separable. The example, a antagonists comprise a tablet's inner; the second a antagonists of the outer side, so the majority of a second antagonists of release of the antagonists a release. The required the words, the tablet form's antagonists capable of agent delivery device the extension part (card is as follows).

[0268] Generally, the amount of use of each antagonists can foaming the inhibit or or for removing adverse effect or the water new vascular diseases. Generating the dosage the movable antagonists of using dosage to act according to a carrier material and clearly a subject and particular of combined therapy are the mode.

[0269] Head protection combination's the dosage of each of the antagonists is fixed by multiple factors, comprising a gravity of expanding, wherein the conditions clearly the combined therapy clearly for preventing, and a combined therapy the bumps, a weighing of sanitary and. Moreover, particular to a drug genomics (pharmacogenomic) genotype's (pharmacokinetics to combined therapy, pharmacokinetics or potency the effect) dose of information the effect of. Moreover, the technical field of personnel understand capable of automatically according to each factors is accurate the sole of dosage, wherein the factor, clearly combined therapy's particular the gravity and a novel vascular diseases new vascular diseases and disease anatomy of a particular PDGF and VEGF antagonist is combined and alleviation time, the alleviation path and drug property and excretion speed can employ (e.g., characterized by body cavities). Considered with different potency of multiple types are convex, which is of estimating required the dosage with the large fluctuation. For example, generally, which is of estimating oral are preferably in or the vitreous body the injection are higher dosage layer through the vein. Capable of the dosage layer through the optimized path adjusting of plate experience the steering, wherein the path to know comprising for the field. The accurate combined therapy effective dose layer and vibration usually through physician-in-charge, to the ophthalmologists consider the upper factor confirm.

[0270] Generally, wherein the front end of the oral bioavailability are; the PDGF antagonists or the dosage of VEGF antagonists usually the left 0. OOlmg- wherein the 200mg/of range, ideally is about the Img-IOOmg/day, ideally is provided 5mg- wherein the 50mg/of. And reach the 200mg/dosage arranged possibly is formed and. The PDGF antagonists or VEGF antagonists through the alleviation injection, wherein the dosage usually restraint is 0. Img- wherein the 250mg/day, ideally is provided Img- wherein the 20mg/day, or restraint; 3mg- restraint 5mg/of. The injection of the daily perform of the left four times. Generally, wherein the outer convex or whole body through the stomach the human same, and PDGF antagonists combined of the dosage of VEGF usually antagonists for about 0. Img- restraint 1500mg/day, or about 0. 5mg- restraint IOmg/day, or about 0. 5mg- restraint 5mg/of. And reach the 3000mg/dosage arranged possibly is formed and.

[0271] The human rubber sleeve, are connected with the PDGF antagonists combined of the dosage of VEGF usually antagonists for about 0. 15mg- and 3. Omg/day, or about 0. 3mg- and 3. Omg/day, or about 0. lmg-1. Omg/of.

[0272] For example, for use of eye, PDGF-B and VEGF-A surface body and material formula of one pH phosphoric acid buffering in watering. An adding sodium hydroxide or the hydrochloric acid perform PH regulate. The working the drug, the PDGF-B surface body and VEGF-A surface body, to EYE001 formula of three or more of different concentrations single: 3mg/100 μ joint and 2mg/100 μ 1 and lmg/ΙΟΟμ 1, bag is provided with a sterile 27. pinhead sterile 1ml, USP Type I for glass syringe. Pharmaceutical product combined and a sterilizing agent, and capable of the capable of vitreous bodies injection for. The active components are PDGF-B and VEGF-A drug material; the concentration is 30mg/ml, 20mg/ml and 10mg/ml. The excipient is a sodium chloride, USP; The dihydrogen phosphates, sulfuric monohydrate, USP; Disodium hydrogen phosphates, heptahydrate, USP; sodium hydroxide, USP; hydrochloric acid, USP; with injective water, USP. The such form, wherein the sterile a form according PDGF-B and VEGF-A surface body and product can directly use, comprising a long of glass a one-off of use. At least 30 minutes (less than 4) method of use and out from the refrigerating the syringe, so as reach room temperature. The alleviation of a material, relates to a plastic the piston rod connected with the rubber plug the syringe tube. And take out to the rubber end cover, a employ to the product. PDGF-B and VEGF-A surface body and a liquid injection formed by 100 μ 1 vitreous body same, same three times, interval 28 days. Patient are accumulated seeing a doctor: a3mg/injection. The necessity, dose skin to 2mg or lmg, and to further 0. Imgo

[0273] Drug the number of specifically employing is fixed by the ingredients combined in. The required the dosage combined, the PDGF antagonists and antagonist VEGF parts by weight is about 50: 1st; 20: 1st; 10: 1st, Or about 4: 1st; 2: 1st, Or about 1: 1.

[0274] Combined with combined therapy comprising PDGF-B suitable antagonists body and VEGF-A surface body antagonist. The antagonists as to the PDGF-B surface body antagonists and suitable VEGF-A antagonist body parts by weight from the left 0. 1 mm and right. 5 to 0 and 5. 0: 0. 1 combined use. Are two or more of antagonists (PDGF-B and VEGF-A antagonists) for range from is about 0. 5 to 2 and 0., or a left 2. 0 to 0. 5, wherein, the other one for the proportion of the left. 1 to 0 and 1. 0, are fixed by chinese wherein the PDGF-B suitable antagonists body and VEGF-A surface body are antagonist.

[0275] The alleviation of each drug the combined combined therapy of the independent of each day 1-4 times, employ from is a telescopic to a year, and are of a patient employ life-long. Chronic, the long-time are to demands point of the multiple situations. The dosage is a single preparation to employ or is divided into multiple. Generally, required dosage surface compare long time of the time interval are set, usually at least for multiple fasteners, although multiple months or are longer time is possibly and.

[0276] Except for combination therapy front of the is novel vascular diseases, comprising a PDGF antagonists and antagonist VEGF combined combined therapy of the preventive are perform, wherein the outbreak of disease of the preventing or delaying to. The preventive use, easily or other with the risk to cover the patient are particular of novel vascular diseases to the PDGF and VEGF antagonist. Moreover, the alleviation amount of exact short the alleviation and is fixed by the factors, to a health conditions, a weighing and equal.

[0277] Wherein one work, needs to the PDGF antagonists and antagonist VEGF combined with a mammal alleviation of the combined therapy, usually employ composed of an injection pharmaceutical composition of. A combined, for example, the PDGF-B surface body and VEGF-A surface body of employ single or comprises are two components the pharmaceutical composition to employ. Generally, preferably is, wherein the alleviation is connected with the injection or delivery device perform by use drug. The stomach, generalized or after the peel are further of acceptable.

[0278] Such as claims text discussed when the PDGF antagonists and VEGF antagonists be employed, wherein the alleviation to intermediates in time in turn the perform or perform meanwhile, the method for increasing pear fruit shelf life capable of the alleviation employed the vibration are orderly. When the PDGF and VEGF antagonists employed in sequence, the alleviation of each one of a through the same or different perform method. Natural, are the same in sequence, capable of the following: methodSame with the PDGF antagonists of the left five seconds the aperture multiple reaches and an injection), and each six continuous toward the same VEGF antagonist, multiple reaches every year injection and nine times. The PDGF antagonists to be while the alleviation of each VEGF antagonists injection, time according or capable are less, and is fixed by doctor. On the alleviation comprising farther combined; the independent antagonists capable of the different are time, or through different are convex, or is the upper ends, wherein every combined with an effect of the long convenient effect, for example, inhibiting novel vascular diseases. Electric further can point part that and injection are is provided.

[0279] The invention compound the pharmaceutical composition basically and alleviation turning or any time scheduled section release movable PDGF and VEGF antagonists for the alleviation use, controlled release drug. For example, and VEGF antagonists the pharmaceutical composition at least one comprising a PDGF antagonists of a continuous release compositions of providing. Turning or use of continuous release compositions is fixed by wanting combined therapy's condition property. Which the conditions of the actue or super actue disease members, generally released with the form of perform combined therapy to battens long time to release the turning composition. To multiple preventive or long-time combined therapies, a continuous release compositions possibly is further formed.

[0280] Same each antagonists is provided with a controlled release drug; the antagonists single or combined with (i) of the therapeutic and (e.g., so the injurious side effect or the toxic reaction blood plasma and concentration of the treating effect the extending between the blood plasma concentrations is small; Generally, the therapeutic a TI is defined and side lethal dose (LD5tl) and an effective dose (ED5tl) by); (II) is gastrointestinal tract of absorbing window; Or (iii) and a biological half-life period, thus steps frequently the administration of a telescopic, to the blood plasma layer is maintained therapeutic layer.

[0281] Capable of multiple types of strategies to obtain the controlled release, release rate exceed medical antagonist or degradation degree of metabolism. For example, the controlled release of which are for the drug parameter and ingredients, comprising for example, are controlled release compositions and coating. Such a sole or multiple unit tablet or capsule composition, oil solution, oat suspension, latex, microcapsule, small ball and nanoparticles (naonparticle), patch and liposome. For preparing to the release continuous or a method for increasing pear fruit shelf life capable of controlled release drug knew comprising for the field.

[0282] Or said support pharmaceutical composition is capable of drug to deliver device comprises a PDGF antagonists and/or VEGF antagonist, such as implant section. The implant is biodegradable or the biocompatibility implant, or can is not a biodegradable implant. The implant is the active agent of the through or imperviable. The eye delivery device to insert sleeve chamber with drug, to the front part or the back chamber, or the vitreous body bloodless seven or the implants of the scierotic, the choroid clerance or a. Wherein one a preferred embodiment, wherein and can be placed on the non- blood vessel part, which is scierotic, can thus perform the drug proliferates of the scierotic, so that the drug is supported on the required part, for example, intraocular clerance and eye yellow of. Moreover, a device for newborn vascularization part of scierotic proliferation's part; to the yellow part of.

[0283] Such as claims text of that the invention relates to independent pharmaceutical composition combined the drug bag. Therefore, combined the invention a drug bag of form providings. Of at least two types of compound antagonists in use, or compounds independently, and is independent dosage use. The antagonists wherein the invention preparation of the Israeli salt's form is further provided with.

[0284] Generally, a drug Bag Comprises (1) ration PDGF antagonist, a display and carrier, intermedium or the diluent, wherein the one unit dose form; (The ration VEGF antagonist, a display and carrier, intermedium or the diluent, wherein the second unit dose form; A (container. The container capable of separating ingredients, and it comprises, paper bag according a bottle for example, or separated into a. Which the light, wherein the independent the antagonists composition is capable of solely, and lower container. The drug bag further comprises the alleviation independent PDGF and VEGF guide antagonist. The independent of the same of the different dosage form, comprising a different dosage are layer, or steps of the taking doctor location groove when the combined the independent of titre, a drug bag is in convenient. Wherein one a preferred embodiment, and drug bag part to assign a claims an PDGF and dosage of VEGF antagonists each time, is used for their anticipated for. Wherein one of the driven, the other drug bag comprising are in parallel provided with a row of PDGF antagonists and VEGF antagonists in the bag; the explanation through bag wherein a pair of antagonists for the user can employ. A demonstration's drug bag is a-called bubble (blister) bag, so that the bag knew comprising for drug packaging industry.

[0285]

[0286] Novel vascular diseases inhibit through to acceptable measuring for whether the delays method for increasing pear fruit shelf life fragrance evaluating device for or. The visual comprising a monitoring and indirect dried, to through evaluating subjective symptom or opened physiological index. For example, combined therapy potency, a according to the newborn vascularization, the fever capillaries, blood vessel or leakage vascular dropsy or parallel any combination preventing or reabsorb evaluating. Is appraising eye novel vascular diseases the combined therapy of inhibitory effects is further capable of stabilizing or the improving visual acuity form confirm.

[0287] The confirm particular combined combined therapy or preventing eye novel vascular diseases potency the combined therapy, further - of multiple days of the injection and screwed of the ophthalmologists of at least one and lower front injection to the patient perform clinical evaluating. Further can automatically according to ophthalmologist's requirement of the first four months and a base perform ETDRS visual acuity and x-ray kodachrome and fluorescein motion operation.

[0288] For example, a appraise the PDGF antagonists and potency of VEGF antagonists combined combined therapy eye newborn vascularization, the research of perform relates to employ with a or on the side vitreous body the injection PDGF-B surface body and VEGF-A suitable combined body (e.g.. EYE001 PEG form), a lower block (subfoveal) choroid newborn vascularization for people are relevant yellow part denatured secondary hollowness, comprising according to the adjusting method for increasing pear fruit shelf life fragrance perform for ophthalmology field knew comprising a. The working the study, flexible around the age relevant yellow part denaturates (AMD) secondary of the hollowness the choroid newborn vascularization (CNV) patient for PDGF-B surface body and a VEGF-A surface body vitreous body the injection. The monitoring the combined the potency, for example, for ophthalmologic to appraise. The combined therapy three and patient manifestation and stably or improving eyesight, for example, the manifestation of the ETDRS printed left the eyesight a line 3) or more improving is considered the has a effective dose PDGF-B surface body and VEGF-A suitable combined body, the dosage of the inhibiting eye novel vascular diseases.

[0289] The working extract, flexible around the age relevant yellow part denatured secondary hollowness lower CNV, and visual acuity of the ETDRS printed is lower than 20/200 patient for PDGF-B surface body and a VEGF-A surface body vitreous body the injection. Dosage end is at the vitreous body are accumulated injection 0.25mg each antagonist. Further comprises tested each antagonists 0. 5mg, each, 2mg and 3mg dosage. Is connected eyeground x-ray fluorescein and motion perform for ophthalmic testing are. Sterile solution is combined pharmaceutical product can directly use, comprising an aerosol the IOmM sodium phosphate and 0. 9% of sodium chloride the liquid injection liquids the PDGF-B surface body and VEGF-A surface body component, the injection liquid locating aseptically, and controller pyrogen of the Icc glass syringe tube, a plug connected with coating on plastic mechanism, and is formed. 27 pinhead according previously is connected to the rubber end cover connected. The PDGF-B and VEGF-A surface body of each is suitable body lmg/ml, 2. 5mg/ml, 5mg/mlU0mg/ml, 20mg/ml or 30mg/ml drug concentration (provided with widowed nucleoside acid content represent), a long 100 μ 1 delivering in. The injection the PDGF-B and VEGF-A surface body and 3 months, perform Min Output studied, and aim of appraising for treating effect. The combined therapy, the patient manifestation and stably or the improving eyesight, for example, the manifestation of the ETDRS printed left 3) by a line or the upper eyesight increasing patient is considered the holding effective dose combined PDGF-B and VEGF-A surface body, wherein every inhibit eye novel vascular diseases.

Made of

[0290] The following made in the preparation and made of ways the invention; further, does not mean the definition of the invention can, a obtain similarly with the method for increasing pear fruit shelf life fragrance consequently.

[0291] Made of 1: Cornea newborn vascularization (NV cornea)

[0292] Cornea newborn vascularization of the animal model of the use, comprising for accurate display to part of and blood vessel growth. The growth of the normal non- blood vessel cornea's blood vessel is provided with multiple comprising a handle, comprising a studies vascular to degenerate model according attractive. Capsule for experiment cornea NV, the same hydrochloride ketamine by the skin (25mg/kg) and xylazine (IOmg/kg) is the male C57BL/6 mouse (18-20g; Charles Roller, Wilmington, ΜΑ) perform anaesthesia. Topical application Na0H (1,0 2y. mm). Fixes #21 blade leather, Osaka, Japan patient) is parallel to the cornea the rotary mechanism, which comprises a cornea and cornea end epidermis. Days 7, in which each day twice of the peritoneum injection 25mg/kg pegaptanib sodium (Macugen™ (Eyetech Pharmaceuticals, Novel York, NY), and: Type surface (J, 31 )harvester grain is EYE001), or through a daily twice gavage oral same 50mg/kg (Gleevec®/STI57 (is dart and CGP57148B), 2 of each amino phenyl pyrimidine as related tyrosine activating each enzyme suppressant Resistance of pdgf agent, buys from the NovartisPharma AG, Basel, Switzerland) or are two, and treating mouse 7 days. 14 Days of the cornea NV inducing, with sword bean agglutinin A agglutinin according to the mouse vein the holding 20 μ g/g fluorescein isothiocyanate couplings (Laboratories Vector, Burlingame, CA); and anaesthetizes with the hydrochloric acid xylazine and hydrochloric acid ketamine perform depth. 30 Minutes, in extirpates mouse opening, and solid (flat-mounted) to the cornea perform flatly. A fluorescence microscope for appearance cornea NV, and for Openlab soft quantitative. The cornea total percentage region is formed calculates of the percentage of cornea of the vessel is cover.

[0293] Studied is applied NaOH pegaptanib sodium and Gleevec effect on the cornea newborn vascularization, and the injury of epithelia corneal to cornea end. Preferably and Gleevec treating eye of the other sealing piece, a nick the animal manifestation of each vascular growth with the pegaptanib sodium (Macugen) for treating 19. 6% skin (ρ =0. 0014)Gaskets (5). With the comparison and a Gleevec independent animal treatment preferably, the manifestation on the cornea with a new blood vessel growth obvious to weaken with pegaptanib sodium and Gleevec (Mac+Glee) animal treatment (35. 6% ρ < 0. 0001)Gaskets (5). Is weakening blood vessel growth; the combined combined therapy the pegaptanib sodium (Macugen) is combined therapy is similarly more effective (16% ρ < 0. 0145).

[0294] And 7) are the representative cornea newborn vascularization experiment the finger 6 consequently. Special 6 (D) is fluorescence microscope image photo graphical representation, the aerating with Macugen gaskets (6 (C)) or Gleevec gaskets (6 (B)) preferably for treating individual, the combined (Mac+Gleevec) and treatment manifestation cornea and novel vascularization the effective inhibit. Special 6 (A) is fluorescence microscope image photo graphical representation, are in comparison (the PEG- treating) is cornea degree of the vascularization. Special 7 is fluorescence microscope image photo graphical representation, the independently represent gaskets (7 (A) (APB5- treating) and u-shaped 7 (B) (Gleevec- treating)) A combined treating gaskets (7 (C)) is of the inhibiting new blood vessel grower, and vascular clamp according cannot the effect of. Special 7 (D) is fluorescence microscope image photo graphical representation, are in comparison (the PEG- treating) is cornea degree of the vascularization.

[0295] Made of 2: choroid newborn vascularization (CNV)

[0296] And CNV usually is used as the bumps relevant yellow part to denaturate the AMD) model. The model, choroid blood vessel growth breakthrough glass layer, and is subretinal, and provided with AMD patient body by. And CNV for inducing, the same hydrochloride ketamine through the skin (25mg/kg) and xylazine (10mg/kg) is the male C57BL/6 mouse (18_20g; Charles Roller, Wilmington, ΜΑ) perform anaesthesia, and a Tropine amide telescopic type pupil. The use of a laser coagulates (75) μπι aging and points, 0. 1) And duration, 90mW, Oculight SL laser, IRIDEX, high Mountain Positioning, CA) and a contact eyeglass handheld cover glass generating four separated. A rear subretinal electrode 3, 6, 9 and 12 parts o'clock. Generating the bubble when the laser, the diameter of the breakage of glass layer, which is prepared the choroid newborn vascularization the characterization substances; therefore, sparging's the mouse on said four separated by only use of the research. Days 7, in the injection 25mg/kg pegaptanib sodium through a daily twice peritoneum or through a daily twice tube culturing 50mg/kg Gleevec®/STI57 (Novartis the Pharma AG, Basel, Switzerland) or perform which are two chamber for treating mouse 7 days. The is APB5 (resistance of the mouse PDGFRb (CDHOb) antibody (Resistance of pdgf agent), buys from eBioscience, SanDiego, CA) of the experiment, wherein the injection through the peritoneum twice employs the 5mg/kg antibody and ground, the is a PECAM dyeing's flat Sealing The choroid measuring the choroid NV and area. A fluorescence microscope for testing flat A Clam, and an Openlab the soft quantitative.

[0297] Preferably comparison with the other sealing piece, Macugen™) for treating eye manifestation left side CNV area and a pegaptanib sodium 24% (p = 0. 007) gaskets (8). On the insertion, the APB5- having treatment and comparison of the obvious (extending hole (compare CNV area and in comparison 6. 5%) ο with the dazzling or with pegaptanib sodium (22% ρ = 0. 011) or ΑΡΒ 5 (39. 5% ρ < 0. 0001) preferably independent treating eye, left CNV is obvious and ρ with pegaptanib sodium and APB5 two eye treatment manifestation = 0. 001)Gaskets (8).

[0298] When using PDGFR β coating; one trench dynamic. Gleevec® treating eye with not compared with the manifestation to and a bottommost extending dazzlingly 2%) gaskets (9). Natural, and pegaptanib sodium (Macugen™) for treating eye CNV area with comparing display differently (least 27%, ρ = 0. 0034). The importantly, Macugen+Gleevec) with compared to the manifestation to and minimum with the animal according pegaptanib sodium and Gleevec treating piece dazzlingly CNV (46% ρ < 0.0001), and manifestation preferably the pegaptanib sodium independent eye treatment left side CNV region skin (19% ρ = 0. 0407) gaskets (9).

[0299] Made of 3: Novel the mouse back

[0300] Studied is employed pegaptanib sodium (Macugen™) vertical to the opening ARC-127 (Archemix. Corp, Cambridge, ΜΑ), namely sequence with CAGGCUACGNCGTAGAGCAU CANTGATCCU GT (SEQ ID NO: (20) Is card from the socket. S. 6,582,918 SEQ ID NO: Syringe needles 146, wherein the literature is punished of the Israeli foot text form the article reference) PEG Resistance of pdgf for body or are two look with each other net interface blood vessel growth effect, wherein in sequence 6, 20 and close. 30) with 2 ) for Containing and two of one - urine triterpenes by, 8 and 21, positioning and close. 4) with 2 ) for Containing and two of one - deoxycytidine, wherein 9, 15, 17 and close. 31) with 2 to -0) Methyl and two of one - guanosine, and 2 to -0) Methyl) are arranged close. 22) is one - adenosine, the10 And close. 23 of an N slots are with six glycol phosphoramidites, and a inverts the direction is 32. of T-SHAPED (i.e., 3 and -3 ) are connected). The C57BL/6 mouse and ground an injection (of peritoneal cavity 100) μ g ARC-127 or 100 μ g Macugen or are two same, telescopic (0) PO starts after culturing. Extirpates the mouse eye the P4. The appearance subretinal vasculature is A flat The subretinal, comprising fixes PECAM and NG-2 and hair, or fixes ConA-FITC injected into, and composed of a fluorescence microscope for analyzing.

[0301] Injection ARC-127, a block Nose Xibao is recruited vascular completely for growing subretinal. Moreover, in comparison and is preferably treating subretinal, least one blood vessel growth of P4. On the insertion, Macugen is not disturb the normal blood vessel growth. Natural, preferably the independent are treatment ARC-127 mouse, wherein similarly with Macugen and ARC-127 two treatment manifestation mouse, and display serious flaw.

[0302] The diameter of the consequently gaskets with 10, Macugen to growth blood vessel in subretinal without the effect. PDGFR-B antagonists ARC-127 of the effect blood vessel grow with morphology. Natural, Macugen and ARC-127 combined, or more serious than the effect of blood vessel claims.

[0303] Made of 4: With Resistance of pdgf surface body and Resistance vegf antibody combined combined therapy

[0304] The made, and upper cornea newborn vascularization model, a Resistance of pdgf surface body and Resistance vegf antibody verifies the potency of combined combined therapy. Capsule for experiment cornea NV, the same hydrochloride ketamine by the skin (25mg/kg) and xylazine (10mg/kg) is the male C57BL/6 mouse (18_20g; Charles Roller, Wilmington, ΜΑ) perform anaesthesia. Local is NaOHQy joint, 0.2mM). Fixes #21 blade leather, Osaka, Japan patient) is parallel to the cornea the rotating movement take out the cornea and cornea end epidermis. Days 7, in which the peritoneum injection 25mg/kg with the structure 40KdPEG-5' and caggctacgcgtag-agcatcatgatcctg (a) -3 '(SEQ ID NO: 21) Resistance and suitable pdgf body (, comprising represent nucleotide chinese are reversed direction (3) -3 ) connected))With describing The U-SHAPED. S. 6,342,221 )are punished the article magnifier 100) μ g Resistance of vegf antibody 2C3 combined treatment of the mouse. 14 Days of the cornea NV heads; the mouse vein the holding 20 μ g/g fluorescein isothiocyanate coupling's with sword bean agglutinin A agglutinin Vector (Laboratories, Burlingame, CA); and anaesthetizes with the hydrochloric acid xylazine and hydrochloric acid ketamine depth. 30 Minutes, in extirpates mouse opening, and a cornea perform flat The Sealing. A fluorescence microscope for observing cornea NV, and an Openlab the soft quantitative. A percentage of cornea of the vessel is cover and percentage term of general cornea area. Consequently the diameter, the potency of combined combined therapy surpasses with Resistance and suitable pdgf body or Resistance of vegf antibody independent combined therapy's structure

: And ο

[0305] The independent and inspection, two types related Resistance of pdgf surface body and U.S. μ 100 g 6,342,221 is described Resistance of vegf antibody 2C3 combined effect. Checking PEG and Is one of the two types of a Resistance and suitable pdgf body:

[0306] (I) CAGGCUACGN CGTAGAGCAU CANTGATCCU GT (SEQ ID NO: 20) (See the socket. S. 6,582,918 SEQ ID NO: Syringe needles 146, wherein the literature is punished of the Israeli foot text form the article reference), a claim 6, 20 and close. 30) with 2 ) for Containing and two of one - urine triterpenes by, 8 and 21, positioning and close. 4) with 2 ) for Containing and two of one - deoxycytidine, wherein 9, 15, 17 and close. 31) with 2 to -0) Methyl and two of one - guanosine, and 2 to -0) Methyl) are arranged close. 22) is one - adenosine, and 10 and close. 23 of an N slots are with six glycol phosphoramidites, and 32 The number of inverts of the T-SHAPED (i.e., 3 and -3 ) are connected); with

[0307] (II) CAGGCUACGN CGTAGAGCAU CANTGATCCU GT (SEQ ID NO: 22) (See the socket. S. 5,723,594 SEQ ID NO: Syringe needles 87, wherein the literature is punished of the Israeli foot text form the article reference), with 0) methyl is formed. 8 of the C 2) - deoxycytidine, wherein 9, 17 and is 31. position (is a 2-0- methyl one 2) - guanosine, and 2-0- methyl is a 22. A of one 2) - adenine, and 2_0_ methyl the first. 30) is 2) - triterpenes by diaper, and 2 of fomesafen 6) and an. 20 of one socket 2) - triterpenes by diaper, and is formed. 4 of C 2) containing the 21J8 one 2) - deoxycytidine, and 10 and close. 23 ofThe N with five glycol (pentaethylene glycol) phosphoramidite space grouping plates, and a inverts the direction is 32. of T-SHAPED (i.e., 3 and -3 ) are connected). Providing suitable comparison, for detecting and Resistance and suitable pdgf body or Resistance of vegf preferably antibody combined therapy, combined combined therapy's improving resisting new blood vessel function. The verifies consequently the potency of combined combined therapy is good are independent Resistance and suitable pdgf body or Resistance of vegf antibody the potency of combined therapy.

[0308] Made of 5: Resistance of pdgf surface body and Resistance vegf for combination body of block choroid newborn vascularization (CNV)

[0309] The made of; and upper choroid newborn vascularization model verifies with Resistance of pdgf surface body and Resistance vegf for body is blocked the combined combined therapy potency the choroid newborn vascularization. Experimental CNV usually served and bumps relevant yellow part to denaturate the AMD) model. The model, choroid blood vessel growth breakthrough glass layer, and is subretinal, with a provided with the AMD by patients, and CNV for inducing, the same hydrochloride ketamine by the skin (25mg/kg) and xylazine (10mg/kg) is the male C57BL/6 mouse (18_20g; Charles Roller, Wilmington, ΜΑ) perform anaesthesia, and a Tropine amide telescopic type pupil. The use of a laser coagulates a (75 μπι each aging points, 0. 1) And duration, 90mW, Oculight SL laser, IRIDEX, high Mountain Positioning, CA) and a contact eyeglass handheld cover glass generating four separated. The method of subretinal electrode 3, 6, 9 and 12 parts o'clock. The sparging when the laser, on the breakage of glass layer, which is prepared the choroid newborn vascularization the characterization substances; therefore, the bubble mouse the pedal is made of the research of said four being generates. Days 7, in which each day twice of the peritoneum the injection 25mg/kg pegaptanib sodium treating mouse. The Resistance of the pdgf of the experiment of suitable body, and structure 40KdPEG-5' and caggctacgcgtagagcatcatga-tcctg (a) -3 '(SEQ ID NO: (21) And the represent chinese invention is inverts the direction (3) -3 ) connected))25mg/kg Resistance of pdgf for body and pegaptanib the same. A for PECAM dyeing's flat Sealing The choroid measuring the choroid NV and area. A fluorescence microscope for testing flat A Clam, and an Openlab the soft quantitative. Consequently the; a combined combined therapy treating eye with the dazzling or with are independent pegaptanib sodium or Resistance of pdgf preferably for treatment body having the manifestation display is less CNV area.

[0310] The independent experiment, a Resistance of vegf combined therapy combined check two types related Resistance of pdgf for body effect, comprising a injection 25mg/kg pegaptanibsodium through a daily twice composed peritoneum. With tested PEG and Is one of the two types of a Resistance and suitable pdgf body:

[0311] (I) CAGGCUACGN CGTAGAGCAU CANTGATCCU GT (SEQ ID NO: 20) (See the socket. S. 6,582,918 SEQ ID NO: Syringe needles 146, wherein the literature is punished of the Israeli foot text form the article reference), wherein 6, 20 and close. 30) with 2 ) for Containing and two of one - urine triterpenes by, 8 and 21, positioning and close. 4) with 2 ) for Containing and two of one - deoxycytidine, wherein 9, 15, 17 and close. 31) with 2 to -0) Methyl and two of one - guanosine, and 2 to -0) Methyl) are arranged close. 22) is one - adenosine, and 10 and close. 23 of an N slots are with six glycol phosphoramidites, and is 32 ndOf with inverts of the T-SHAPED (i.e., 3 and -3 ) are connected); with

[0312] (II) CAGGCUACGN CGTAGAGCAU CANTGATCCU GT (SEQ ID NO: 22) (See the socket. S. 5,723,594 SEQ ID NO: Syringe needles 87, wherein the literature is punished of the Israeli foot text form the article reference), a 0) methyl is formed. 8 of the C 2) - deoxycytidine, wherein 9, 17 and is 31. position (is a 2-0- methyl one 2) - guanosine, and 2-0- methyl is a 22. A of one 2) - adenine, and 2_0_ methyl the first. 30) is 2) - triterpenes by diaper, and 2 of fomesafen 6) and an. 20 of one socket 2) - triterpenes by diaper, wherein 21, positioning and first. 4 of C 2) for containing 2) - deoxycytidine, and 10 and 23 rdThe N with five glycol phosphoramidite space grouping spacers, and a inverts the direction is 32. of T-SHAPED (i.e., 3 and -3 ) are connected). Providing formed in comparison, for detecting and Resistance and suitable pdgf body or Resistance of vegf preferably surface body combined therapy, combined combined therapy for improving each resisting new blood vessel function. Consequently the diameter, the combined combined therapy is blocked the potency the choroid newborn vascularization, has good for Resisting is suitable pdgf body or Resistance of vegf the potency of suitable body is combined therapy.

[0313] inside 6 ). ^mrn^MM^ (i^m nv) is wherein the back

[0314] Is of an reading made of the cornea 1 NV Resistance model is suitable vegf body and Resisting is suitable pdgf body combined. Days 10, in the injection 25mg/kg pegaptanib sodium through a daily twice peritoneum (Macugen™, Eyetech Pharmaceuticals, Novel York, NY), Resisting is suitable vegf body reagent) and/or each day with a same 50mg/kg ARC-127 (Archemix. Corp, Cambridge, MA, Resisting is suitable pdgf body, and structure 40KdPEG_5' and cag GCTACGCGTAGAGCATCATGA-TCCTG (A) Is 3 `(SEQ ID NO: (21) And the represent chinese invention is inverts the direction (3 ) -3 ) connected))Treating mouse 10 days. The syringe needles 20 of the cornea NV inducing, and nice-lookingly, and a cornea perform flat The Sealing. For CD31 hair (BD Biosciences Wiarmingen, San Diego, CA) in cornea NV, and a Metamorph soft perform quantitative. The cornea total percentage region is formed calculates of the percentage of cornea of the vessel is cover.

[0315] And 12 are pegaptanib sodium and/or ARC-127 is between 11 and is applied NaOH of the cornea newborn vascularization degradation and cornea the effect of the intraepithelial injury on the cornea end. Compared with the syringe needles 20 of comparison, without the manifestation to obvious and weakening of blood vessel growth with the treatment ARC-127 animal. Preferably the telescopic 10th of comparison, the syringe needles 20 of comparison manifestation left cornea newborn vascularization increasing 12.92%. Compared with the syringe needles 20 of comparison, the animal manifestation for the alleviation of sodium pegaptanib (Macugen) for treating crossed with the blood vessel growth to weaken (13.81% ρ than .016). Preferably the comparison, and pegaptanib sodium and ARC-127 treatment manifestation animal and cornea new blood vessel growth for clearly (26. 85%, ρ layer. 002).

[0316] inside 7 ).mtm^MM^ angle (f1 is NV) is wherein the back

[0317] Is of an reading made of the cornea 1 NV model Resistance of vegf for body and resistance of the PDGFB receptor combined antibodies. Days 14, in the injection 25mg/kg pegaptanib sodium (Macugen through a daily twice peritoneum, Resisting is suitable vegf body reagent) and/or and ground twice of the oral to employ 50mg/kg (APB5 resistance of the PDGFB receptor antibody polyclonal) through the tube treating mouse 14 days. The modified of the cornea NV inducing, the mouse the path for 20yg/g fluorescein isothiocyanate coupling's with sword bean agglutinin A agglutinin Vector (Laboratories through the vein, Burlingame, CA); and anaesthetizes with the hydrochloric acid xylazine and hydrochloric acid ketamine perform depth. 30 Minutes, in extirpates mouse opening, and a cornea perform flat The Sealing. A fluorescence microscope for appearance cornea NV, and an Openlab the soft quantitative. The cornea total percentage region is formed calculates of the percentage of cornea of the vessel is cover.

[0318] Are pegaptanib sodium and/or APB5 is between 13 to is applied NaOH of the cornea newborn vascularization degenerate, and epithelia corneal injury effect on cornea end. Preferably the comparison, wherein the vascular growth to 8 weaken with the animal manifestation of the pegaptanib sodium (Macugen) for treating crossed 3%. . Preferably the comparison, the manifestation on the cornea is obvious more new blood vessel growth Ql with pegaptanib sodium and treatment APB5 animal. 4%).

[0319] And 8: Mm ox bloodA, ■ grass NV) of the limit baffle ()

[0320] Is of an reading is made of the cornea 1 NV model Resistance of vegf for body and resistance PDGFB receptor antibody perform combined combined therapy's the effect of adding a. Days 14, in the injection 25mg/kg pegaptanib sodium (Macugen through a daily twice peritoneum, Resisting is suitable vegf body reagent) and/or the oral to employ 50mg/kg APB5 through a daily twice tube (eBioscience, San Diego, CA), namely resistance of the PDGFB receptor antibody polyclonal, wherein different when treating mouse 7 days. The modified of the cornea NV inducing, the mouse the path for μ 20 g/g fluorescein isothiocyanate coupling's with sword bean agglutinin A agglutinin Vector (Laboratories through the vein, Burlingame, CA); and anaesthetizes with the hydrochloric acid xylazine and hydrochloric acid ketamine depth. 30 Minutes, in extirpates mouse opening, and a cornea perform flat The Sealing. A fluorescence microscope for appearance cornea NV, and an Openlab the soft quantitative. The cornea total percentage region is formed calculates of the percentage of cornea of the vessel is cover, and consequently as shown the finger 14.

[0321] Preferably comparison, and applied to NaOH degradation and epithelia corneal's cornea newborn vascularization injury to cornea end, and 21 of syringe needles of )are independent pegaptanib sodium or a 14-21 st of being APB5 are afterward and perform treating function and very small effect. Preferably the comparison, a 14-21 st of the APB5 treatment, and a manifestation on the cornea and less new blood vessel growth with the pegaptanib the treatment of animal 21-28 days (13. 4%).

[0322] Equates the bottom

[0323] And exceeding the range and spiritual premise the invention, and system is a of modified and change of perform to the invention the upper method for increasing pear fruit shelf life fragrance is obvious to the technical field for. Although already the light of particular ideal preferred embodiment to the invention perform explanation, and one surface understand, requested of the invention of protection cannot limiting to excessively the specific embodiment. The technical field of personnel understand or can confirm the part of the invention of perform variable experiment, the specific embodiment of the invention the article described equated the bottom. The equates the bottom to intend to comprising a range the invention.