Anticoagulant peptides.

0001Anticoagulants are useful therapeutic agents in the pharmacological treatment of, for example, acute deep venous thrombosis, pulmonary embolism, acute arterial embolization of the extremities, myocardial infarction, and disseminated intravascular coagulation. Proplylactic administration of anticoagulants is believed to prevent a recurrance of embolism in patients with rheumatic or arteriosclerotic heart disease and to prevent certain thromboembolic complications of surgery. Administration of anticoagulants has also been indicated in the treatment of coronary artery and cerebrovascular disease. Artrial thrombosis, particularly in arteries supplying the heart muscle and brain, is a leading cause of death.

0002Hirudin is a 65 residue polypeptide isolated from the salivary glands of leeches. It is an anticoagulant agent, which is a thrombin specific inhibitor. Although quite potent, clinical use of hirudin isolated form leech extracts seems unlikely because of its limited quantity, expense and allergic reactions which commonly follow administration of any foreign protein of this size.

0003Applicants have discovered a specific region of hirudin that is responsible, at least in part, for its anticoagulant activity. This region has been chemically synthesized and certain of its analogs appear to bind to the recognition site of thrombin but not the enzymatic cleavage site which is spatially separate. Binding of the synthetic peptides competitively prevents binding of the fibrinogen to the recognition site of thrombin, a prere­quisite to fibrin production and clot formation. The peptides of this invention possess significant anticoagu­lant activity and their unusual ability to bind only to the recognition site without binding to the cleavage site of thrombin may allow for a scientifically interesting and therapeutically significant adjunct to anticoagulant therapy.

0004Peptide derivatives of the formula X-A₁-A₂-A₃-A₄-A₅-A₆-A₇-A₈-A₉-A₁₀-Y wherein X is an amino terminal residue selected from hydrogen, one or two alkyl groups of from 1 to 6 carbon atoms, one or two acyl groups of from 2 to 10 carbon atoms, carbobenzyloxy or t-butyloxy carbonyl; A₁ is a bond or is a peptide containing from 1 to 5 residues of any amino acid; A₂ is Phe, SubPhe, β-(2- and 3-thienyl)alanine, β-(2-and 3-furanyl)alanine, β-(2-, 3-, and 4-pyridyl)alanine, β-(benzothienyl-2- and 3-­yl)alanine, β-(1- and 2-naphthyl)alanine, Tyr or Trp; A₃ is Glu or Asp; A₄ is any amino acid; A₅ is Ile, Val, Leu, Nle, or Phe; A₆ is Pro, Hyp, 3,4-dehydroPro, thiazolidine-­4-carboxylate, Sar, NMePgl or D-Ala; A₇ is any amino acid; A₈ is any amino acid; A₉ is a lipophilic amino acid selected from Tyr, Tyr(SO₃H), Trp, Phe, Leu, Nle, Ile, Val, Cha and Pro or is a dipeptide containing at least one of these lipophilic amino acids; A₁₀ is a bond or is a peptide fragment containing from one to five residues of any amino acid; and Y is a carboxy terminal residue selected from OH, C₁-C₆ alkoxy, amino, mono- or di-(C₁-C₄) alkyl substituted amino, or benzylamino; are useful anticoagulant agents.

0005The following common abbreviations of the amino acids are used thoughout this specification: Cha - cyclohexylalanine Orn - ornithine Glt - glutaryl Mal - maleyl NpA - beta-(2-naphthyl)alanine Gly - glycine Ala - alanine Val - valine Leu - leucine Ile - isoleucine Pro - proline Phe - phenylalanine Trp - tryptophan Met - methionine Ser - serine Thr - threonine Cys - cysteine Tyr - tyrosine Asn - asparagine Gln - glutamine Asp - aspartic acid Glu - glutaminc acid Lys - lysine Arg - arginine His - histidine Nle - norleucine Hyp - hydroxyproline 3,4-dehydroPro - 3,4-dehydroproline Tyr(SO₃H) - tyrosine sulfate Pgl - phenylglycine NMePgl - N-methyl-phenylglycine Sar - sarcocine (N-methylgycine) pSubPhe - para substituted phenylalanine SubPhe - ortho, meta, or para, mono- or di-­substituted phenylalanine DAla - D-alanine Ac - acetyl Suc - succinyl pClPhe - para-chloro-phenylalanine pNO₂Phe - para-nitro-phenylalanine

0006An alkyl group and the alkyl portion of an alkoxy group is taken to include straight, branched, or cyclic alkyl groups, for example, methyl, ethyl, propyl, isopro­pyl, butyl, isobutyl, <u style="single">tert</u>-butyl, pentyl, isopentyl, <u style="single">sec</u>-­pentyl, cyclopentyl, hexyl, isohexyl, cyclohexyl and cyclopentylmethyl. An acyl group of from 2 to 10 carbon atoms is taken to include straight, branched, cyclic, saturated and unsaturated acyl groups having 1 or 2 carbonyl moieties per group, for example, acetyl, benzoyl, maleyl, glutaryl and succinyl. A halogen group is a fluoro, chloro, bromo or iodo group.

0007The term "any amino acid" as used herein includes the naturally occurring amino acids as well as other "non-­protein" α-amino acids commonly utilized by those in the peptide chemistry arts when preparing synthetic analogs of naturally occurring peptides. The naturally occurring amino acids are glycine, alanine, valine, leucine,isoleu­cine, serine, methionine, threonine, phenylalanine, tyro­sine, tryptophan, cysteine, proline, histidine, aspartic acid, asparagine, glutamic acid, glutamine, arginine, ornithine, and lysine. Examples of "non-protein" α-amino acids are norleucine, norvaline, alloisoleucine, homoargi­nine, thiaproline, dehydroproline, hydroxyproline (Hyp), homoserine, cyclohexylglycine (Chg), α-amino-n-butyric acid (Aba), cyclohexylalanine (Cha), aminophenylbutyric acid (Pba), phenylalanines substituted at the ortho, meta, or paraposition of the phenyl moiety with one or two of the following a (C₁-C₄) alkyl, (C₁-C₄) alkoxy, halogen, or nitro groups or substituted with a methylenedioxy group, β-2- and 3-thienylal- anine, β-2- and 3-furanylalanine, β-­2-, 3-,and 4-pyridylalanine, β-(benzothienyl-2- and 3-­yl)alanine, β-(1- and 2-naphthyl)alanine, O-alkylated derivates of serine, threonine, or tyrosine, S-alkylated cysteine, the O-sulfate ester of tyrosine, 3,5-­diiodotyrosine and the D-isomers of the naturally occurring amino acids.

0008The term "lipophilic amino acid" includes Tyr, Phe, Leu, Nle, Ile, Val, His and Pro.

0009The natural amino acids with the exception of glycine, contain a chiral carbon atom. Unless otherwise specifi­cally indicated, the optically active amino acids, referred to herein, are of the L-configuration. For example, any of the amino acids of the A₁ or A₁₀ group can be of the D- or L-configuration. As is customary, the structure of peptides written out herein is such that the amino terminal end is on the left side of the chain and the carboxy terminal end is on the right side of the chain.

0010The polypeptides of formula 1 can form pharmaceuti­cally acceptable salts with any non-toxic, organic or inorganic acid. Illustrative inorganic acids which form suitable salts include hydrochloric, hydrobromic, sulphu­ric and phosphoric acid and acid metal salts such as sodium monohydrogen orthophosphate and potassium hydrogen sulfate. Illustrative organic acids which form suitable salts include the mono, di and tricarboxylic acids. Illustrative of such acids are, for example, acetic, glycolic, lactic, pyruvic, malonic, succinic, glutaric, fumaric, malic, tartaric, citric, ascorbic, maleic, hydro­xymaleic, benzoic, hydroxybenzoic, phenylacetic, cinnamic, salicylic, 2-phenoxybenzoic and sulfonic acids such as methane sulfonic acid and 2-hydroxyethane sulfonic acid. Salts of the carboxy terminal amino acid moiety include the non-toxic carboxylic acid salts formed with any suit­able inorganic or organic bases. Illustratively, these salts include those of alkali metals, as for example, sodium and potassium; alkaline earth metals, such as calcium and magnesium; light metals of Group IIIA includ­ ing aluminum; and organic primary, secondary and tertiary amines, as for example, trialkylamines, including triethylamine, procaine, dibenzylamine, 1-ethenamine, N,Nʹ-dibenzylethylenediamine, dihydroabietylamine, N-­(lower)alkylpiperidine, and any other suitable amine.

0011As will any generic group of chemical compounds, certain groups are preferred. Applicants prefer those peptide derivatives of formula 1 wherein X is hydrogen, acetyl, or succinyl. Also preferred are those formula 1 compounds wherein A₁ is -His-Asn-Asp-Gly-Asp-; -Asn-Asp-Gly-Asp-; -Asp-Gly-Asp-; -Gly-Asp-; -Asp-; Thr-Pro-Lys-Pro-Gln-Ser-His-Asn-Asp-Gly-Asp; Thr-Pro-Asn-Pro-Glu-Ser-His-Asn-Asn-Gly-Asp; Ser-His-Asn-Asp-Gly-Asp, or a bond. A₂ is preferably Phe, β-2- or 3-thienylalanine, Tyr, Trp, or pClPhe, Npa; A₃, Glu; A₄, Glu, Asp, Pro or Ala; A₅, Ile, Leu; A₆, Pro, Sar, DAla, Hyp or NMePgl; A₇, Glu, Asp, Ala, Gln; A₈, Glu, Asp, Ala; A₉, Pro, Ala-Tyr, Ala-Cha, Tyr-Cha, Tyr-Leu, Ala-Phe, Tyr-Tyr; A₁₀, Glu, Asn, Asp-Glu, Pro, Gln, Ala, a bond, D-Lys, Lys, D-Asp or Orn; Y, OH or NH₂. Especially preferred are those peptide derivatives of formula 1 wherein either X is acetyl and A₁ is Gly-Asp or Asp or X is succinyl an A₁ is a bond and wherein A₂, is Phe; β-(2-thienylalanine) or Tyr; A₃, Glu; A₄, Glu or Pro; A₅, Ile; A₆, Pro; A₇, Glu or Gln. A₈, Glu or Asp; A₉, Tyr-Leu, Ala-Tyr; Tyr-Tyr, Ala-Phe, Ala-Cha, or Pro; A₁₀,Gln, Asp, Pro, a bond, Asp-Glu, D-Asp, D-Lys or D-Glu; and Y, OH or NH₂.

0012The proteins of this invention can be prepared by a variety of procedures readily known to those skilled in the art. Such procedures include the solid phase sequen­tial and block synthesis, gene cloning and combinations of these techniques. The solid phase sequential procedure can be performed using established automated methods such as by use of an automated peptide sythesizer. In this procedure an α-amino protected amino acid is bound to a resin support. The resin support employed can be any suitable resin conventionally employed in the art for the solid phase preparation fo polypeptides, preferably poly­styrene which has been cross-linked with from 0.5 to about 3 percent divinyl benzene, which has been either chloro­methylated or hydroxymethylated to provide sites for ester formation with the initially introduced α-amino protected amino acid.

0013An example of a hydroxymethyl resin is described by Bodanszky et al., <u style="single">Chem. Ind. (London) 38</u>, 1597-98 (1966). A chloromethylated resin is commercially available from Bio Rad Laboratories, Richmond, California, and the prep­aration of such a resin is described by Stewart et al., "Solid Phase Peptide Synthesis" (Freeman &amp; Co., San Fran­cisco 1969), Chapter 1, pp. 1-6. The protected amino acid can be bound to the resin by the procedure of Gisin, <u style="single">Helv. Chem Acta, 56</u>, 1476 (1973). Many resin bound, protected amino acids are commercially available. As an example, to prepare a polypeptide of this invention wherein the car­boxy terminal end is a Thr residue, a tert-butyloxycar­bonyl (Boc) protected Thr bound to a benzylated, hydroxy­methylated phenylacetamidomethyl (PAM) resin can be used and is commercially available.

0014Following the coupling of the α-amino protected amino acid to the resin support, the protecting group is removed using any suitable procedure such as by using trifluoro­acetic acid in methylene chloride, trifluoroacetic acid alone, or HCl in dioxane. The deprotection is carried out at a temperature of between 0°C and room temperature. Other standard cleaving reagents and conditions for removal of specific α-amino protecting groups may be used. After removal of the α-amino protecting group the other amino protected amino acids are coupled step-wise in the desired orther. Alternatively, multiple amino acid groups may be coupled by the solution method prior to coupling with the resin supported amino acid sequence.

0015The α-amino protecting group employed with each amino acid introduced into the polypeptide sequence may be any such protecting group known in the art. Among the classes of α-amino protecting groups contemplated are (1) acyl type protecting groups such as: formyl, trifluoroacetyl, phthalyl, toluenesulfonyl (tosyl), benzenesulfonyl, nitro­phenylsulfenyl, tritylsulfenyl, o-nitrophenoxyacetyl and α-­chlorobutyryl; (2) aromatic urethan type protecting groups such as benzyloxycarbonyl and substituted benzyloxycar­bonyl, such as p-chlorobenzyloxycarbonyl, p-nitrobenzyl­carbonyl p-bromobenzyloxycarbonyl, p-methoxybenzyloxy­carbonyl, 1-(p-biphenylyl)-1-methylethoxycarbonyl, α, α-­dimethyl-3,5-dimethoxybenzyloxycarbonyl and benzhydryloxy­carbonyl; (3) aliphatic urethan protecting groups such as tert-butyloxycarbonyl (Boc), diisopropylmethoxycarbonyl, isopropyloxycarbonyl, ethoxycarbonyl and allyloxycarbonyl; (4) cycloalkyl urethan type protecting groups such as cyclopentyloxycarbonyl, adamantyloxycarbonyl and cyclo­hexyloxycarbonyl; (5) thio urethan type protecting groups such as phenylthiocarbonyl; (6) alkyl type protecting groups such as triphenylmethyl (trityl) and benzyl; and (7) trialkylsilane groups such as trimethylsilane. The preferred α-amino protecting group is tert-butyloxycar­bonyl.

0016The selection of an appropriate coupling reagent is within the skill of the art. A particularly suitable coupling reagent where the amino acid to be added is Gln, Asn or Arg is N,Nʹ-diisopropylcarbodiimide and 1-hydroxy­benzotriazole. The use of these reagents prevents nitrile and lactam formation. Other coupling agents are (1) carbodiimides (e.g., N,Nʹ-dicyclohexylcarbodiimide and N-­ethyl-Nʹ-(y-dimethylaminopropylcarbodiimide); (2) cyana­mides (e.g., N,N-dibenzylcyanamide); (3) ketenimines; (4) isoxazolium salts (e.g., N-ethyl-5-phenyl-isoxazolium-3ʹ-­sulfonate; (5) monocyclic nitrogen containing heterocyclic amides of aromatic character containing one through four nitrogens in the ring such as imidazolides, pyrazolides, and 1,2,4-triazolides. Specific heterocyclic amides that are useful include N,Nʹ-carbonyldiimidazole and N,N-­carbonyl-di-1,2,4-triazole; (6) alkoxylated acetylene (e.g., ethoxyacetylene); (7) reagents which form a mixed anhydride with the carboxyl moiety of the amino acid (e.g., ethylchloroformate and isobutylchloroformate) or the symmetrical anhydride of the amino acid to be coupled (e.g., Boc-Ala-O-Ala-Boc) and (8) nitrogen containing heterocyclic compounds having a hydroxy group on one ring nitrogen (eg., N-hydroxyphthalimide, N-hydroxysuccinimide and 1-hydroxybenzotrizole). Other activating reagents and their use in peptide coupling are described by Kapoor, <u style="single">J. Pharm. Sci., 59,</u> pp. 1-27 (1970). Applicants prefer the use of the symmetrical anhydride as a coupling reagent for all amino acids except Arg, Asn and Gln.

0017Each protected amino acid or amino acid sequence is introduced into the solid phase reactor in about a four-­fold excess and the coupling is carried out in a medium of dimethylformamide: methylene chloride (1:1) or in dimethylformamide alone or preferably methylene chloride alone. In cases where incomplete coupling occurs, the coupling procedure is repeated before removal of the α-­amino protecting group, prior to the coupling of the next amino acid in the solid phase reactor. The success of the coupling reaction at each stage of the synthesis is monitored by the ninhydrin reaction as described by E. Kaiser et al, <u style="single">Analyt. Biochem. 34</u>, 595 (1970).

0018After the desired amino acid sequence has been ob­tained, the peptide is removed from the resin. This can be done by hydrolysis such as by treatment of the resin bound polypeptide with a solution of dimethyl sulfide, p-­cresol and thiocresol in dilute aqueous hydrofluoric acid.

0019As is known in the art of solid phase peptide synthesis many of the amino acids bear functionalities requiring protection during the chain preparation. The use and selection of the appropriate protecting group is within the ability of those skilled in the art and will depend upon the amino acid to be protected and the pre­sence of other protected amino acid residues on the peptide. The selection of such a side chain protecting group is critical in that it must be one which is not removed by cleavage during cleavage of the protecting group of the α-amino moiety. For example, suitable side chain protecting groups for lysine are benzyloxycarbonyl and substituted benzyloxycarbonyl, said substitutent being selected from halo (e.g., chloro, bromo, fluoro) and nitro (e.g., 2-chlorobenzyloxycarbonyl, p-nitrobenzyloxy­carbonyl, 3,4-dichlorobenzyloxycarbonyl), tosyl, t-­amyloxycarbonyl, t-butyloxycarbonyl and diisopropyl­methoxycarbonyl. The alcoholic hydroxy group of threo­nine an serine can be protected with an acetyl, benzoyl, tert-butyl, trityl, benzyl, 2,6-dichlorobenzyl, or benzyl­oxycarbonyl group. The preferred protecting group is benzyl.

0020These groups can be removed by procedures well known in the art. Typically protecting group removal is done after the peptide chain synthesis is complete but the protecting groups can be removed at any other appropriate time.

0021The anticoagulant dose of a peptide derivative of this invention is from 0.2 mg/kg to 250 mg/kg of patient body weight per day depending on the patient, the severity of the thromobotic condition to be treated and the peptide derivative selected. The suitable dose for a particular patient can be readily determined. Preferably from 1 to 4 daily doses would be administered typically with from 5 mg to 100 mg of active compound per dose.

0022Anticoagulant therapy is indicated for the treatment and prevention of a variety of thrombotic conditions, particularly coronary artery and cerebrovascular disease. Those experienced in this field are readily aware of the circumstances requiring anticoagulant therapy. The term "patient" used herein is taken to mean mammals such as primates, including humans, sheep, horses, cattle, pigs, dogs, cats, rats and mice.

0023Although some of the peptide derivatives may survive passage through the gut following oral administration, applicants prefer non-oral administration, for example, subcutaneous, intravenous, intramuscular or intraperi­toneal; administration by depot injection; by implant preparation; or by application to the mucous membranes, such as, that of the nose, throat and bronchial tubes, for example, in an aerosol can containg a peptide derivative of this invention in a spray or dry powder form.

0024For parentral administration the compounds may be administered as injectable dosages of a solution or sus­pension of the compound in a physiologically acceptable diluent with a pharmaceutical carrier which can be a sterile liquid such as water and oils with or without the addition of a surfactant and other pharmaceutically acceptable adjuvants. Illustrative of oils which can be employed in these preparations are those of petroleum, animal, vegetable, or synthetic origin, for example, peanut oil, soybean oil, and mineral oil. In general, water, saline, aqueous dextrose and related sugar solu­tions, ethanol and glycols such as propylene glycol or polyethylene glycol are preferred liquid carriers, parti­cularly for injectable solutions.

0025The compounds can be administered in the form of a depot injection or implant preparation which may be formu­lated in such a manner as to permit a sustained release of the active ingredient. The active ingredient can be com­pressed into pellets or small cylinders and implanted sub­cutaneously or intramuscularly as depot injections or implants. Inplants may employ inert materials such as biodegradable polymers or synthetic silicones, for example, Silastic, silicone rubber manufactured by the Dow-Corning Corporation.

EXAMPLES

0026This invention is illustrated by the following, nonlimiting examples.

EXAMPLE 1

Preparation of H-Gly-Asp-Phe-Glu-Glu-Ile-Pro-Glu-Glu-­Tyr-Leu-Gln-OH

0027The peptide was snythesized by solid-phase methods using 0.1 mmol of a 0.66 mmol/g Boc-Gln-PAM resin. Double symmetrical anhydride couplings were performed with 2.0 mmol Nα-Boc-amino acid (Peptides International) except in the case of Boc-Gln, which was coupled by the DCC/HOBT method. The side chain protection utilized was: Asp(Chx), Glu(Bzl), Tyr(2-BrZ). Upon completion of the synthesis the Nα-Boc protection was removed with 50% trifluoroacetic acid in methylene chloride. The resin was washed three times with methylene chloride, neutralized with three washings of 10% diisopropylethylamine in methylene chloride, washed three times with methylene chloride, and dried <u style="single">in</u><u style="single">vacuo</u>.

0028The peptide was deprotected and cleaved from the resin with HF containing 2% anisole at 0°C, for 35 min. The HF was removing <u style="single">in vacuo</u> at 0°C, the peptide precipita­ted with ethyl ether, extracted from the resin with 30% aqueous acetic acid and lyophilized.

0029The peptide was purified by desalting on a 92 x 2.6 cm Sephadex G-15 column in 5% aqueous acetic acid and lyophilized. Preparative HPLC was performed on a C¹⁸ Vydac 218TP1010 (250 x 10 mm) column with 24% acetonitrile in 0.1% aqueous trifluoroactic acid at 5 ml/min. The major peak was collected and lyophilized leaving 101 mg of the desired produce (58% yield based on initial resin substi­tution). Homogeneity was determined by HPLC and TLC. HPLC Vydac 218TP54 (250 x 4.6 mm) C¹⁸ column, 2 ml/min, t₀ = 1.9 min: time of elution with a 15-40% acetonitrile in 0.1% trifluoroacetic acid linear gradient at 1%/min. (HPLC) is 14.4 min.

0030TLC: [Merck 5715 20 x 20 cm Silica gel 60 plates (0.25 mm thickness)] n-butanol/acetic acid/water/pyridiene (6:1.2:4.8:6) (TLC I) Rf = 0.42; isopropanol/conc. ammonium hydroxide/water (3:1:1) (TLC II) Rf = 0.32; n-­butanol/acetic acid/water (4:5:5) (TLC III) Rf = 0.70. FAB-MS: (M + H) - 1468.4 ± 1 mµ (calcd. 1468.6). Amino acid analysis: (6N HCl hydrolysis; 24 hr. at 106°C). Asx 1.03 (1); Glx 5.05 (5); Pro 1.03 (1); Gly 1.00 (1); Ile 0.97 (1); Leu 1.01 (1); Tyr 0.93 (1); Phe 0.98 (1); NH₃ 1.06 (1). ε 280 = 1254. 85% peptide content by weight.

0031In the same manner, the peptides of the following example 2-32 were prepared.

EXAMPLE 2

0032Ac-Ser-His-Asn-Asp-Gly-Asp-Phe-Glu-Glu-Ile-Pro-Glu-­Glu-Tyr-Leu-Gln-OH

EXAMPLE 3

0033H-Asn-Asp-Gly-Asp-Phe-Glu-Glu-Ile-Pro-Glu-Glu-Tyr-Leu-­Gln-OH

EXAMPLE 4

0034H-Asp-Phe-Glu-Glu-Ile-Pro-Glu-Glu-Tyr-Leu-Gln-OH

EXAMPLE 5

0035H-Phe-Glu-Glu-Ile-Pro-Glu-Glu-Tyr-Leu-Gln-OH

EXAMPLE 6

0036H-Asp-Phe-Glu-Ala-Ile-Pro-Glu-Glu-Tyr-Leu-Gln-OH

EXAMPLE 7

0037H-Asp-Phe-Glu-Glu-Ile-Pro-Ala-Glu-Tyr-Leu-Gln-OH

EXAMPLE 8

0038H-Asp-Phe-Glu-Glu-Ile-Pro-Glu-Glu-Tyr-Leu-Ala-OH

EXAMPLE 9

0039H-Asp-Phe-Glu-Glu-Ile-Pro-Glu-Glu-Glu-Leu-Ala-OH

EXAMPLE 10

0040Ac-Asp-Phe-Glu-Glu-Ile-Pro-Glu-Glu-Tyr-Leu-Gln-OH

EXAMPLE 11

0041Ac-Gly-Asp-Phe-Glu-Glu-Ile-Pro-Glu-Glu-Tyr-Leu-Gln-OH

EXAMPLE 12

0042Suc-Phe-Glu-Glu-Ile-Pro-Glu-Glu-Tyr-Leu-Gln-OH

EXAMPLE 13

0043H-Asp-Phe-Glu-Glu-Ile-Pro-Glu-Gly-Tyr-Leu-NH₂

EXAMPLE 14

0044H-Gly-Asp-Tyr-Glu-Glu-Ile-Pro-Glu-Glu-Tyr-Leu-Gln-OH

EXAMPLE 15

0045H-Gly-Asp-Trp-Glu-Glu-Ile-Pro-Glu-GLu-Tyr-Leu-Gln-OH

EXAMPLE 16

0046H-Gly-Asp-pClPhe-Glu-Glu-Ile-Pro-Glu-Glu-Tyr-Leu-Gln-­OH

EXAMPLE 17

0047H-Gly-Asp-pNO₂Phe-Glu-Glu-Ile-Pro-Glu-Glu-Tyr-Leu-Gln-­OH

EXAMPLE 18

0048H-Gly-Asp-Phe-Glu-Glu-Ile-Sar-Glu-Glu-Tyr-Leu-Gln-OH

EXAMPLE 19

0049H-Gly-Asp-Phe-Glu-Glu-Ile-DAla-Glu-Glu-Tyr-Leu-Gln-OH

EXAMPLE 20

0050H-Gly-Asp-Phe-Glu-Glu-Ile-Hyp-Glu-Glu-Tyr-Leu-Gln-OH

EXAMPLE 21

0051H-Gly-Asp-Phe-Glu-Glu-Ile-NMePgl-Glu-Glu-Tyr-Leu-Gln-­OH

EXAMPLE 22

0052H-Gly-Asp-Phe-Glu-Pro-Ile-Pro-Glu-Asp-Ala-Tyr-Asp-Glu-­OH

EXAMPLE 23

0053H-Ser-His-Asn-Asp-Gly-Asp-Phe-Glu-Glu-Ile-Pro-Glu-­Glu-Tyr-Leu-Gln-OH.

EXAMPLE 24

0054H-Gly-Asp-Phe-Glu-Pro-Ile-Pro-Glu-Asp-Ala-Tyr-Pro-NH₂

EXAMPLE 25

0055Suc-Phe-Glu-Pro-Ile-Pro-Glu-Asp-Ala-Tyr-Pro-NH₂

EXAMPLE 26

0056H-Asp-Phe-Glu-Pro-Ile-Pro-Glu-Glu-Ala-Tyr-Gln-OH

EXAMPLE 27

0057H-Asp-Phe-Glu-Glu-Ile-Pro-Glu-Glu-Ala-Tyr-Gln-OH

EXAMPLE 28

0058H-Asp-Phe-Glu-Glu-Ile-Pro-Glu-Asp-Ala-Tyr-Gln-OH

EXAMPLE 29

0059Suc-Phe-Glu-Pro-Ile-Pro-Glu-Glu-Tyr-Leu-Pro-NH₂

EXAMPLE 30

0060H-Gly-Asp-β-(2-thienyl)alanyl-Glu-Glu-ILe-Pro-Glu-Glu-­Tyr-Leu-Gln-OH

EXAMPLE 31

0061H-Gly-Ala-O-methyltyrosyl-Glu-Glu-Ile-Pro-Glu-Glu-Tyr-­Leu-Gln-OH

EXAMPLE 32

0062Suc-Phe-Glu-Pro-Ile-Pro-Glu-Glu-Pro-OH

EXAMPLE 33

0063H-Asp-Phe-Glu-Glu-Ile-Pro-Glu-Ala-Tyr-Leu-Gln-OH

EXAMPLE 34

0064H-Asp-Phe-Glu-Glu-Ile-Pro-Glu-Glu-Ala-Leu-Gln-OH

EXAMPLE 35

0065H-Gly-Asp-Phe-Glu-Glu-Leu-Pro-Glu-Glu-Tyr-Leu-Gln-OH

EXAMPLE 36

0066H-Gly-Asp-Phe-Glu-Glu-Val-Pro-Glu-Glu-Tyr-Leu-Gln-OH

EXAMPLE 37

0067H-Gly-Asp-Phe-Glu-Glu-Phe-Pro-Glu-Glu-Tyr-Leu-Gln-OH

EXAMPLE 38

0068H-Asp-Phe-Glu-Pro-Ile-Pro-Glu-Glu-Tyr-Leu-Gln-OH

EXAMPLE 39

0069Suc-Phe-Glu-Pro-Ile-Pro-Glu-Glu-Tyr-Leu-Gln-OH

EXAMPLE 40

0070H-Gly-Asp-Phe-Glu-Pro-Ile-Pro-Glu-Asp-Ala-Tyr-Glu-OH

EXAMPLE 41

0071H-Gly-Asp-Phe-Glu-Glu-Ile-Pro-Glu-Asp-Ala-Phe-Asp-Glu-OH

EXAMPLE 42

0072Ac-Phe-Glu-Glu-Ile-Pro-Glu-Glu-Tyr-Leu-Orn-NH₂

EXAMPLE 43

0073H-Gly-Asp-Phe-Glu-Pro-Ile-Pro-Glu-Glu-Tyr-Tyr-Glu-OH

EXAMPLE 44

0074H-Asp-Phe-Glu-Pro-Ile-Pro-Glu-Glu-Tyr-Tyr-Gln-OH

EXAMPLE 45

0075Suc-Tyr-Glu-Pro-Ile-Pro-Glu-Glu-Ala-Phe-Lys-NH₂

EXAMPLE 46

0076Suc-Tyr-Glu-Pro-Ile-Pro-Glu-Glu-Ala-Phe-D-Lys-NH₂

EXAMPLE 47

0077Suc-Tyr-Glu-Pro-Ile-Pro-Glu-Glu-Ala-Phe-Orn-NH₂

EXAMPLE 48

0078Suc-Tyr-Glu-Pro-Ile-Pro-Glu-Glu-Ala-Tyr-Lys-NH₂

EXAMPLE 49

0079Suc-Tyr-Glu-Pro-Ile-Pro-Glu-Glu-Ala-Phe-Lys-OH

EXAMPLE 50

0080Suc-Tyr-Glu-Pro-Ile-Pro-Glu-Glu-Ala-His-Lys-OH

EXAMPLE 51

0081Suc-Tyr-Glu-Pro-Ile-Pro-Glu-Glu-Ala-Cha-Lys-NH₂

EXAMPLE 52

0082Suc-Tyr-Glu-Pro-Ile-Pro-Glu-Glu-Leu-Phe-Lys-NH₂

EXAMPLE 53

0083Suc-Tyr-Glu-Pro-Ile-Pro-Gln-Glu-Ala-Phe-Lys-NH₂

EXAMPLE 54

0084Suc-Tyr-Glu-Pro-Ile-Pro-Glu-Glu-Ala-Phe-Glu-OH

EXAMPLE 55

0085Suc-Tyr-Glu-Pro-Ile-Pro-Glu-Glu-Ala-Phe-Gln-OH

EXAMPLE 56

0086Suc-Tyr-Glu-Pro-Ile-Pro-Glu-Glu-Ala-Cha-Asp-OH

EXAMPLE 57

0087H-Gly-Asp-Phe-Glu-Pro-Ile-Pro-Glu-Asp-Ala-Tyr-D-Asp-OH

EXAMPLE 58

0088Ac-HV-1_45-65

0089The peptides of examples 2-58 have the following properties: <tables id="tabl0001" num="0001"><img file="EP0276014A2_D0001.tif" /></tables><tables id="tabl0002" num="0002"><img file="EP0276014A2_D0002.tif" /></tables><tables id="tabl0003" num="0003"><img file="EP0276014A2_D0003.tif" /></tables><tables id="tabl0004" num="0004"><img file="EP0276014A2_D0004.tif" /></tables><tables id="tabl0005" num="0005"><img file="EP0276014A2_D0005.tif" /></tables><tables id="tabl0006" num="0006"><img file="EP0276014A2_D0006.tif" /></tables><tables id="tabl0007" num="0007"><img file="EP0276014A2_D0007.tif" /></tables><tables id="tabl0008" num="0008"><img file="EP0276014A2_D0008.tif" /></tables>