PT93320B - PROCESS FOR THE PREPARATION OF THERAPEUTIC PEPTIDEES - Google Patents

Description

APPLICANT: EPÍGRAFE: INVENTORS:

DESCRIPTION OF INVENTION PATENT No. 93 320 η ADMINISTRATORS OF THE TULANE EDUCATIONAL FUND, North American (State of Louisia), industrial headquarters at 1430 Tulane Avenue. New Orleans, Louisiana 70122, United States of America. " PROCESS FOR THE PREPARATION OF THERAPEUTIC PEPTIDES "

Claim of right of priority under Article 4 of the Paris Convention of 20 March 1883. 1NPI MOD 113 RF 18732 i i

King ': PTC / Oiomeasure-3ombesin Cirs. 5-7; 9; The present invention relates to a process for the preparation of therapeutic peptides and quot;

10

Mod. 71 -10000 ex. · 89/07 20

A process for the preparation of a linear (i.e., non-cyclic) bombesin of biologically active mammalian gastrin releasing peptide (PLG) and amphibian bombesin, with an active site and a binding site responsible for the binding of peptide to a receptor of a target cell; the cleavage of a peptide bond at the naturally occurring bombesin active site or PLG is not required for in vivo biological activity. The analogue has one of the following modifications: a) a deletion of a residue within the active site and a modification of a residue outside the active site, or b) a substitution of one or two residues within the site activated with a synthetic amino acid. The analog is capable of binding and acting as a concurrent inhibitor of the naturally occurring peptide by binding to the receptor, and, due to one of the modifications, fails to exhibit the in vivo biological activity of the naturally-produced peptide. 25 35 GJL.So4 Ref: PTC / 3iomeasure-Bombesin CIPS. 5-7: 9; 10

5

Mod. 71 -10000 ex. - 89/07 20 25

DESCRIPTION OF THE OBJECT OF THE INVENTION THAT ADMINI3TÍUT0R5. 0Γ ΊΙΙΕ TULANE EDUCATIONAL FUND, North American (State of Louisiana), industrial based at 1430 Tulbne Avenue, New Orleans, Louisiana 70122, United States of America, wishes to obtain in Portugal, for: " PROCESS FOR THE PREPARATION OF THERAPEUTIC PEPTIDE "; The present invention relates to a process for the preparation of therapeutic peptides, which may be used, e.g., in the treatment of benign or malignant tissue and disordered tissue proliferation. gastrointestinal disorders. The amphibian peptide bcmbesin, pGlu-Gln-Arg-Leu-Gly-Asn-Gln-Trp-Ala-Val-Gly-His-Leu-Met-> H10 (Anastasi et al., Experientia 27 (166-167 1971), is closely related to the mammary gastrin releasing peptides (CPLG), eg, porcine FLG, N-Ala-Pro-Val-Ser-Val-Gly-Gly-Gly-Thr-Val-Leu-Ala -Lys-Met-Tyr-Pro-Arg-Gly-Asn-His-Trp-Ala-Val-Gly-IIis-Leu-1α- (NH)) (Mcdonald et al., Diochem.Niophys. , 90: 227-233 (1979) and human PLG H2N-Val-Pro-Leu-Pro-Ala-Gly-Cly-Cly-Thr-Val-Leu-Thr-Lys-Î ± -Tetr-Pro-Arg -Gly-Pis-Trp-Ala-Val-rly-Lys -Leu-I-et (NHO). Bombesin has been reported to be a growth factor for several human cancer cell lines, including lung carcinoma cells (CPCP), and has been found in human breast and prostate cancer (Unveman et al., eds., Recent Resulte in Cancer Research - Teptide Hormones in Lung Cancer, Springer-Verlag, New York: 19S6). It is known that several of these cancers secrete hormone in peptides related to PLG or bombesin. Accordingly, brombladder antagonists have been suggested as agents for the treatment of these cancers.

Cuttitta et al. demonstrated that a mono-clonal bombesin antibody inhibited in vivo the growth of a small cell line from xenograft lung cancer to nude mice (Cuttitta 30 10 15

Mod. 71-10000 ex.-89 / 07-2025

n. oo

Ilef: PTC / Qicmeasure-Bombesin CIPS. 5-7; 9; 10 et al., Cancer Survey 4: 707-727 (1985)). In murine 3T3 fibroblasts that react to the bombosin nitazole effect, Zachary and Rozengurt observed that a β antagonist substance (Spantida) was acting as a bombesin antagonist (Zachary et al., Proc. Natl Acad Sci. (US), S2: 7616-7620 (1985)). Heinz-Srian et al. they replaced Ilis at position 12 in bombesin by D-Fhe and observed bombesin antagonist activity in dissociated nuclei from guinea-pig pancreas (Llez-Erian et al., Am.

Physiol. 252, G139-G442 (1987)). Rivier described a study developed directly for the restriction of the freedom of conformation of the bioactive C-terminal decapeptide C of bombesin, by the incorporation of intramolecular disulfide bridges; however, Rivier has mentioned that to date the bombesin analogs with this modification do not exhibit any antagonistic activity (rivier et al., " Competitive Antagonists of Pepti de Honnones ", in Abstracts of the International Symposium on Bombesin Analog Peptides in Health and Disease, Rome (Italy) (October, 19S7) Abbreviations (rare): pGlu = H "C-CH-C00H (pyroglutamic acid);

2I I c II 0

Nle = COOH (norleucine) (ca2) 3-al2

3-pyridyl-alanine β-leu-β-homoleucine Y -leu = gamma-homleucine D-Cpa-Dp-chlorophenylalanine Ilypro = hydroxyproline Nal = naphthylalanine Sar = sarcosine Sta (statin) = (3S, 4S) -4- amino-3-hydroxy-6-methylheptanoic acid, and has the chemical structure:

Ref: PTC / Biomeasure - Bombosin CIPS. 5-7; 9; 10 \ /

CH

I

CII "OH

2-yl) -NH-CII-CH-COH; 2 2 2 AHPPA = (3S, 4S) -4-amino-3-hydroxy-5-phenylpentanoic acid ACIIPA = 10

(33,4S) -4-amino-5-cyclohexyl-3-hydroxypentanoic acid R = right configuration (D); S = left configuration (L); racemate = equal to the mixture of R and S-1-methyl-Tlis; 3-methyl-His = methyl group (CH 2) on nitrogen at or 3 positions of Histidine:

CK2-Al

Mod. 71-10000 ex. - 89/07 20 25

CH-N

NHg --- CH --- COOH

Linear (i.e., non-cyclic) peptide, which is a naturally occurring and biologically active mammalian gastrin-releasing amphibian peptide (PLG) analogue, has an active site and a binding site responsible for the binding of the peptide to a receptor in a target cell, cleavage of a peptide bond at the active site of naturally occurring bombesin or PLG for in vivo biological activity is unnecessary, the analog having one of the following modifications; an elimination of a residue within the active site and a modification of a residue outside the active site, or (b) a substitution of one or two residues within the active site by a synthetic acid araine, the analog is capable of acting as a concurrent inhibitor of the peptide produced naturally by means of receptor binding, and due to one of the modifications, can not exhibit the in vivo biolagic activity of the naturally-produced peptide. Locations of the antagonist modifications are determined by the location of the active site on the peptide.

Mod. 71-10000 ex. -89/07 20 25 30 61,864 Hef: PTC / Biomeaaure-Bombesin CIPS. 5-7; 9; 10

yl. 1 * | / " J

produced naturally. For example, linear peptides for which the introduction of a non-peptide bond between two residues, or the replacement of two natural amino acids by a β-amino acid or the deletion (" des " of the C-terminal residue are useful to create or to give the antagonist activity, are those in which the activity is associated with the two C-terminal residues of the amino acid chain. Thus, the active site of the naturally occurring peptide of which the peptides according to the invention are analogous preferably includes an amino acid at least at the carboxy terminus in the medium of the peptide and the linear peptide according to the invention includes that amino acid at its carboxy terminus. Likewise, where the active site is located at the amino terminal portion of the naturally-produced peptide, the corresponding analogs of the invention have modifications at their amino terminal moieties. In preferred embodiments, the active site includes at least one amino acid residue located at the carboxyl-terminal half of the naturally occurring and biologically active peptide, and that amino acid residue is located at the carboxyl-terminal half of the linear peptide. In further preferred embodiments, the active site includes at least one amino acid residue located at the amino terminus of the naturally occurring peptide, and that amino acid residue is located at the amino terminus of the linear peptide. Modifications may be made in a zone enclosed in the receptor ligand, or in a nonbinding zone. Preferably, the analogs of the invention are homologous in 25%, and most preferably homologous in 50% of the naturally produced peptides. The analogs of the invention may have one of the modifications indicated in the generic formulas set forth below; or a non-peptide bond in place of a peptide bond between an amino acid of the active site and a contiguous amino acid; or a statin residue or AIIPPA or AClPA, or a p- or -amino acid instead of one or two natural amino acids; or an elimination of the C-terminal amino acid which may or may not be accompanied by the addition of a substituent on the actual C-terminal group or the presence of an N-terminal residue which is not the terminal amino acid

Mod. 71-10000 ex. · 89/07 20 25 G1.8C4 nef: PTC / Hiomeasure-Bombesin CIPS. 5-7; 9; 10

N gives the peptides from which the analogs are derived. (Statin, AHPPA, and ACIIPA have the chemical structures defined above. Where statin is employed in this case, AHPPA or ACIIPA may also be employed.). Non-peptide bond means that the carbon atom which participates in the bond between two residues is reduced from a carbonyl carbonyl to a methylene carbon, ie, Cl -NH; or, less preferably, the residue attached to the carbon atom has a sulfur atom or a methylene carbon instead of its amino group, ie CII-S or CO-CH 2. (A detailed discussion of the chemistry of non-peptide bonds has been published by Coy et al. (1988) Tetrahedron 44, 3, 385-841, incorporated herein by reference). A modification of the naturally-occurring peptide to create an antagonist is the amino terminus of the molecule, which may be an electron donor residue, for example a nitrogen-containing amino acid, for example those described for amino terminal positions in the genomic formulas indicated below; for example the N-terminal amino acid, which is A or, if A ° is eliminated, is A1 or, if A ° and are eliminated, is A1 below, it may be an aromatic D-isomer, or may be an alkylated amino acid . (Where " D " is not indicated as the configuration of an amino acid is meant L.). Another modification is the C-terminal residue, which may be any one of the " V / " described below. The invention includes a therapeutic peptide comprising from seven to nine amino acid residues, the peptide being an analog of one of the following naturally occurring carboxy-terminating peptides with a Met: (a) litorin residue; (b) neuromedin; (c) the ten carboxy-terminal amino acid zone of mammalian gastrin-releasing peptide; and (d) the carboxy terminal ten amino acid zone of amphibian bombesin, the therapeutic peptide having the formula: α 1 .λ 1-α 2-τγρ-λ 4-α 5-λ 5-λ 7 - in which A ° = Cly, Ala,

Het, Vai, β-aminobutyric acid, Gin, Asn, Leu, He, Vat or the D-isomer of any one of pX-Phe (wherein X = F, Cl, 3r, -C13). ϊ64

Ref: PTC / Biomeasure - Bombesine CTPS. 5-7; 9; 10

/ I 10 15

Mod. 71 · 10000 ex. Or II, or CH), Trp, Cys or β-Nal, or is eliminated; A is the D or L isomer of any one of pGlu, Xle, or ananinobutyric acid, or the D isomer of any one of Ala, Val, Gin, Asn, Leu, Ile, Met, pX-Phe Hence X =?, Cl, Br, KG. CII, II or CK2), Trp, TiO, Cys or β-Xal, or is eliminated; 2 A = pGlu, Gly, Ala, Val, Gin, Asn, Leu, Ile, Met, pX-Phe (wherein X = FCl, Br, K00, C-, II or CH3), Trp, -Xal, ilis, 1-methyl-IIis, or 3-methyl-IIis; 4-Ala, Vai, Gin, Asn, Cly, Leu, He, Xle, Amino-Tyrmic Acid, Vet, pX-Phe (which X1, Cl, Br, NC0, OH, II or CII) , Trp, Cys or (3-Xal; 5A = Gin, Asn, Gly, Ala, Leu, Ile, Xle, aminobutyric acid, Met, Yal pX-Phe (wherein X = F, Cl, H or CII "), Trp, Thr or P -Xal; 6.3 A = Sar, Cly is the D-isomer of any one of Ala, Val, Gin, Asn, Leu, Ile, Met, pX-Phe (wherein X = F, Cl, Br, XO ,,, CII, H or CLI), Trp,

Cys or (3-Xal; A = 1-methyl-Nis, 3-methyl-Ilis, or Ilis, provided that if A is present A can not be pGlu; and if A or A is present, it can not be pGlu; and when A ° is eliminated and is pGlu, R3 has to be H and has to be the Glu moiety which forms the imine ring in pGlu, and also since v is pos.

I1 II

-NH-CK-R "-C-V

and is the group identifying any one of the acid anions Gly, Ala, cyclohexyl-C1-4 alkyl, C1-4 alkoxy, C1-4 alkoxy, Ala, Val, Leu, Ile, Ser, Asp, Asn, Glu, Gin, pX-Phe (wherein X = H, F, Cl, Br, NO O O or CH), Trp, Cys, Met, Pro, Hy 30

Pro or isopropyl, cyclohexylmethyl, [D -Xal, {3-naphthylmethyl, or phenylmethyl; and V is 0, R or 5,

N

V -6- 10 15

Mod. 71 -10000 ex. - 89/07 20 25 30 61.SG4 Re f: PTC / B i ome as ure-Good he s i n CIP3. 5-7; 9; 10

wherein R is any of C alquilo alquilo, alkyl, C al al, alkenyl, C 4 al al alkynyl, phenyl, naphthyl, or phenyl-C_10 »alkyl, and wherein R »e is independently any of II, alkyl, C7-10 phenylalkyl, lower acyl, or wherein R1 is any of C1 -C4 alkyl, phenylalkyl, or lower acyl; provided that when one of Π or R is NFIR, the other is O-O-R; and provided that any asymmetric carbon atom may be Π, S or a racemic mixture; and also provided that each R 2 and R 3 independently is II, phenylalkyl, alkyl (in which is C 3 -C 8 alkenyl, alkynyl, phenyl, naphthyl or phenylalkyl), or lower acyl, and R 4 and are attached to N-terminal amino acid of the peptide, and also provided that when one of R or R is COR, the other X 2 1 must be II, or a pharmaceutically acceptable salt thereof. Preferably, the therapeutic peptide has the formula wherein A is = Gly, D-Phe or is deleted; Î ± = β-Glu, D-Phe, D-Ala, D-γ-Nal, D-Cpa or D-Asn; 2A = Gin, His, 1-methyl-His, or 3-methyl-His; 4 Λ = Ala; A5 = Val; A, Sar, Gly, D-Phe or D-Ala; A7-Ilis provided that when R is CH-CH 2, 2 is the group identifying Leu or «ώ ώ 1 Phe; or when R3 is CH0, is the group identifying either -Leu or Leu; or when Π 3 is CIKSI-CH 2, is the Leu identifier group or is isopropyl, cyclohexylmethyl, Î ± -naphthylmethyl or phenylmethyl, provided that V is

N / 5 \

R 2 and each R 2 and R 2 is II. 5 6 -7 · 6 '35 1 10

Mod. 71-10000 ex. -89/07 20 25

.0 .1 2 '4 5 6 A -A -A -Trp-A -A -A G1 · 364

Ref: PTC / Diomeasure-Bombesin CIPS. 5-7; 9; 10

Preferably, the peptide of the general formula wherein V is NIIR is that R and NH. 6

Most preferably, the therapeutic peptide will have the following amino acid formulas; pGlu-Gln-Trp-Ala-Val-Gly-IIis-statine-amide; The present invention also includes a therapeutic peptide comprising from eight to ten amino acid residues, inclusive, the peptide one of which is a peptide of the formula ## STR1 ## wherein R 1 and R 2 are as defined above. analogous to one of the following naturally occurring peptides terminating at the carboxy terminus with a residue: (a) lithium; (b) neuromedin; (c) the ten amino acids of the carboxy terminus of mammary gastrin-releasing peptide; (d) the ten amino acid zone of the carboxy-terminal amphibian pumpin, the therapeutic peptide having the formula:

-W wherein A = Gly, Nle, Î ±, Î ± -aminobutyric, or the D-isomer of any one of Ala, Val, Gin, Asn, Leu, Ile, Met, pX-Phe (wherein X = F, Cl, Br , Νθ,, OH, H or CH₂), Trp, Cys or [3-Nal, or is eliminated; A is the D or L isomer of any one of pGlu, Nle, or Î ± -aminobutyric acid, or the D isomer of any of Ala, Val, Gin, Asn, Leu,

Ile, Met, pX-Phe, pX-Phe (where X = F, Cl, Br, NOg, 011, H or CII>, Trp, Cys, [3-Nal, or is deleted; Gly, Ala, Val, Gin, Asn, Leu, Ile, Met, pX-Phe (wherein X = F, Cl, Br, NOg, II, II, or CH3), Trp, Cys, 1-metyl-His, or 3-methyl-4-Ais, Ala, Val, Gin, Asn, Gly, Leu, Ile, Nle, α-amino butyric, Met, pX-Phe (wherein X = F, Cl, Br, Ng, OH, H or CII), trp, Cye, or (3-Na, A = Gin, Asn, Gly, Ala, Leu, Ile, Nle, σ (aminobutyric acid, Met, Wherein X = F, Cl, Br, OH, II or C10, Trp, The, or (3-Nal; A3) = Sar, Gly, or the D isomer of any one of Ala, Go, Gin, Asn, Leu, -S- 35

10 15 31,864

Ref; PTC / Biomeasure-Bombeain CIPS. 5-7; 9; 10

(Wherein X = F, Cl, Br, NO 2, OH, H or CH 2), Trp,

Cys, or (3-Nal; 7 Λ = 1-methyl-Isis, 3-methyl-His, or Ilis, provided that if A ° is present, A1 can not be pClu; A can not be pGlu and when A is deleted and A is pClu, R 2 has to be II and must be the Glu moiety which forms the imine ring in pGlu, and also since V- can be: 1

Zo 0I2 II

Mod. 71 -10000 ex. - 89/07 20 25

In which R 4 is CH 2 -NH, -CH 2 -CH 2, or CH 2 CH 2, and each of χ and Z 0, independently, and the identifying group of any one of of the amino acids Gly, Ala, Val, Leu, Ile, Ser, Asp, Asn, Cl, Gin, (3-Nal, pX-Phe (wherein X = H, F, Cl, Br, NO2, OH or CII ), trp, Cys,, Tet, Pro, Hypro, cyclohexyl, Ala, or cyclohexylmethyl provided that when R3 is CH2-MI and Z1 is the group identifying any one of the amino acids Gly, Ala, Ala, Leu , Ile, Ser, Asp, Asn, Clu, Gin, pX-Phe (wherein X = II, F, Cl, Br, NO, CT, or Cl * Trp, Cys, Et, Pro, Hypro, or Cyclohexylmethyl , is the sole identifying group of any one of the amino acids Ser, Asp, Cl, Cys, Pro, Hy-Pro, or cyclohexylmethyl, and provided that when R3 is C1-4-NH and R7 is the group identifying a any of the amino acids Gly, Ala, Val, Leu, Ile, Ser, Asp, Asn, Glu, Gin, PX-Phe (wherein X = H, F, Cl, Br, XO, CII or CII) trp, Cys, Met, Pro, or Hypro, Z0 is the only identifying group of any one of the amino acids Ser, Asp, Glu, Cys, Pro, Hypro, or cyclohexylmethyl; and V is CP.R or O>.

N \

Each R,, R,, R e and R,, independently, is II, lower alkyl, phenyl lower alkyl, or naphthyl lower alkyl; and provided that any asymmetric carbon atom may be Π, or a racemic mixture; and also provided that each R 1 and R 2, independently, is H, C 1-12 alkyl, C 7 -C 10 phenylalkyl, COE (wherein is C 1-20 alkyl, C 3-20 alkenyl, C 1-6 alkynyl, naphthyl, C 1-10 alkynyl) or acyl lower alkyl, and R and R0 are attached to the N-terminal amino acid of the peptide, and the peptide X is 9-35 1 5 10 15

Mod. 71-10000ex. -89/07 20 30 61.864 Ref: PTC / Bioneasure-Bombesin CIPS. 5-7; 9; Provided that when one of R6 is COE3, its pharmaceutically acceptable salt. Preferably,

the other has to be Π, or a therapeutic peptide has the template in which = Gly, D-Phe, or is deleted; D-Phe, D-Phe, D-Ala, D- (¾ -Nal, D-Cpa, or D-Asn; 2A = Gin, Ilis, 1-methyl-His, or 3-methyl-His R 4 is Ala, A 5 = Val, Sar, Gly, D-Phe, or D-Ala, A 7 = His, wherein R 5 is C 1-4 -alkyl, each Z 3 is cyclohexylmethyl or is the Leu Most preferably, the therapeutic peptide includes D-Nal at the A-position, wherein each of Z 2 and Z 2 independently is Leu or Phe, or Phe, or Z, is the identifying group of Met, Leu or Phe.

Examples of such peptides are: D-Nal-Gln-Trp-Ala-Val-Gly-His-Leuv1 / CH2NU / Leu-NH2 # or D- (3-Nal-Gln-Trp-Ala-Val-Cly In addition, the therapeutic peptide may include wherein R 2 is CH 2 -NH 2, and the carbon atom attached to Z 1 has the F configuration. An example of such a peptide is D-Phe In addition, the peptide may include wherein A or A is an amino acid, V is OR; or a pharmaceutically acceptable salt thereof, wherein R 1, R 2, R 3 and R 4 are as defined above. The invention also includes a therapeutic peptide comprising from seven to nine amino acid residues in which the amino acid residues of the peptide, , the peptide being an analog of one of the following naturally occurring peptides which terminate at the carboxy terminus with a residue: (a) yltoredine, (b) neuromedin, (c) the ten amino acids of the carboxy terminus of peptide liberating mammary gastrin, and (d) the ten amino acids of the carboxy terminal zone of amphibian bombesin, the therapeutic peptide having the β10.0

Ref: PTC / 3iomeasure-Bombesin CIPS. 5-7; 9; 10 mule:

10 15

Mod. 71 -10000 ex. A5-A5-A6-A7 -W wherein A3 = Gly, Nle, Amino-butyric acid, or the D-isomer of any one of Ala, Val, Gin (Wherein X = F, Cl, CH 3, NO 2, CH 2, H or CH), Trp, Cys, or -Nal, or is eliminated; A is the Î ± or L isomer of any one of Glu, Nle, or Î ± -oct-antinobutyric, or the D-isomer of any Ala, Val, Gin, Asn, Leu, He, Met pX-Phe (wherein X = F, Cl, Br, NO, QII, H or CH), Trp, Cys or N-MJ-Nal), or is eliminated; 2 A = pGlu, Gly, Ala, Val, Gin, Asn, Leu, Ile, Met, pX-Phe (wherein X = F Cl, 3r, NO, OH, R or CH 2), Trp, Cys, β-Ala, Val, Gin, Asn, Gly, Leu, Ile, Nle, Î ± -aminobutyric acid, Met pXF-Nal; β-Ala, β- Phe (wherein X = Γ, Cl, Dr, NO, OH, K or CU), trp, Cys, or ^ A A = Gin, Asn, Gly, Ala, Leu, Ile, Nle, -aminobutyric acid, Met, Val pX-Phe (where X = F, Cl, Dr, OH, Π or Cl), Trp, thr, or Nal; any one of Ala, Val, Cln, Asn, Leu, Ile, Met, pXP Cys, or 13-Nal;

Ile, Met, p-X-Phe (wherein X = F, Cl, Br, NO, OH, H or Cl), Trp, A = 1-methyl-Isis, 3-methyl-nis, or Ilis; Provided A 2 is present, A 2 can not be pGlu; and if A₂ or A₂₂ is present, A can not be pGlu; and when A or A is present, A can not be pGlu; and when Λ is deleted and Λ is pClu, P 2 has to be H and HQ must be the portion of Clu which forms the inine ring in pGlu; and also since V may be; 30

U / 10 -N-CTT-C-N? In which is the identifying group of any one of the amino acids Gly,

Ala, Val, Leu, Ile, Ser, Asp, Asn, Glu, Gin, p-X-Phe (wherein X 1, H, F,

Cl, Br, NC4, OIT or CK3), Trp, Cys, ret, Pr o, or HyPro; and each R1, R2,

Cl. 36-1 Hei; P7C / 3iomeasure-Bombesin CIP3. 5-7: 9: 10

1 5 10 15

Mod. 71 -10000 ex. Independently, is II, lower alkyl, phenyl lower alkyl, or naphthyl lower alkyl; and provided that any asymmetric carbon atom may be Π, s or a racemate; and also since each of the groups is independently selected from the group consisting of hydrogen, C1-6 alkoxy, C1-6 alkenyl, C1-6 alkenyl, C1-6 alkenyl, C1-6 alkenyl, C1-4 alkenyl, C1-4 alkenyl, Are also attached to the N-terminal amino acid of the peptide, and also provided that when one of P. or R0 is COE2, the other has to be H, or a pharmaceutically acceptable salt thereof. Preferably, the peptide includes wherein A = = Gly, Q-Phe, or is deleted; A1 = p -flu, D-Phe, D-Ala, D-Nal, DCpa, or D-AsA; 2A "= Gin, Ilis, 1-methyl-lys, or 3-methyl-His; 4A = Ala; A5 = Val; A = Sar, Gly, D-Phe, or D-Ala; P? - His; provided that Z is the group identifying any one of the amino acids Leu or p-X-Phe D or L (wherein X = Π, F, Cl, Dr, XO_OH or CH); and each R 3 and R 4 independently is H, lower alkyl, phenyl-lower alkyl, or naphthyl-lower alkyl. Most preferably, the peptide includes wherein it is Leu, R8 is H, and each R6e is lower alkyl. Examples <! peptides are: D-Phe-Gln-Trp-Ala-Val-Gly-IIis-Leu-Ethylamide; or D-Phe-G 1 n-Trp-Al a-Vai -C-1 and -1-s-Leu-NH 4. The invention also includes a therapeutic peptide comprising between six and eight amino acid residues, inclusive, the peptide being an analog of one of the following naturally occurring peptides terminating at the carboxy terminus with a Met: (a) litorin residue; (b) neuromedin; (c) the ten amino acids of the carboxy terminal zone of mammalian gastrin-releasing peptide; and (d) the ten amino acids of the carboxy-terminal zone of amphibian bombesin, the therapeutic peptide having the formula:

61,364

Ref: PTC / Biomeasure-Bombesin CIPS. 5-7; 9; 10

R

Wherein A = Gly, Nle, Î ±, Î ± -aminobutyric, or the D-isomer of any one of Ala, Val, Gin, A3n, Leu, Aβ, Î ±, β- Ile, Met, pX-Phe (where X = F, Cl, Br, NO 2, OH, h or CH), Trp, Cys, or (3- Mal, or is eliminated; A = the D or L isomer of a any of pGlu, Nle, or acarbutoxyacetic acid, or the D isomer of any one of Ala, Val, Gin, Asn, Leu, Ile Met, pX-Phe (wherein X = F, Cl, Br, NO2, O, II or CH3), Trp, Cys, or -Nal, or is eliminated;

A = pGlu, Gly, Ala, Val, Gin, Asn, Leu, Ile, Met, p-X-Phe (where X = F

Cl, Br, N0, 011, Π or CH), Trp, Cys, -NNal, Ilis, 1-methyl-Tis, or

Mod. 71 -10000 ex. 3-methyl-IIis; 4A = Ala, Val, Gin, Asn, Gly, Leu, Ile, Nle, aminobutyric acid, Met, pX-Phe (wherein X = F, Cl, Br, NO, OH, Trp, Cys, ort-Nal; 5A = Gin, Asn, Gly, Ala, Leu, Ile, Nle, α-aminobutyric acid, Met, Val, pX-Phe (wherein X = F, Cl, Br, OU, II, or CH), Trp , Tlir, or [3-Nal; 6, JA = Sar, Gly, or the D isomer of any one of Ala, Val, Cln, Asn, Leu, Ile, Met, pX-Phe (wherein X = F, Cl, Br, NO, H, Cl 3), Trp, Cys, or -Nal; A = 1-methyl-His, 3-methyl-His, or Tlis; provided A estiver is present, A * can not be pGlu; and if or A * is present, A2 can not be pGlu; and when A ° is eliminated and A * is pGlu, R has to be H and R 2 has to be the Glu portion which forms the imine ring in pGlu; and also since IV can be: R.

Wherein each R 4, independently, is II, lower alkyl, phenylalkyl, or naphthyllower alkyl; provided that any asymmetric carbon atom may be R 3, or a rac atomic mixture; also wherein each R3 and R7 independently are phenyl,

Mod. 71-10000 ex. -89/07 20 25 30

Ui .-- 0-1? .QI: p7C / i.e. exoestrus-benzoin CIP3. 5-7; 3; 10

(wherein Εχ is alkyl, C, al alalkenyl, phenyl, naphthyl, or phenylC "oualkyl or lower acyl, and R 3-20-20-20 x e e and R os tej are attached to the amino acid terminus of the peptide, G -GFbGH1 dCS1}, wherein when one of R1 or R3 is COE2, the other has to be J1, or a pharmaceutically acceptable salt thereof. Preferably, the therapeutic peptide includes wherein A0 = Gly, D-Phe or is deleted; A1 = p-Glu, D-Phe, D-Ala, D-PnbI, D-Cpa, or D-Asn; 3-methyl-His; A4 = A5 = Val; A = Sar, Gly, D-Phe, or D- / UaA7 = Ilis, wherein each R3 is H; Preferably, when N '0 or h' is other than II, A is Gly, A is Gly, A is V is, A is Ala and A is The invention also includes a therapeutic peptide bombesin having the formula: pGlu-Gln-Arg-Leu-Gly-Asn-Gln-Trp -Ala-Va The invention also includes an effective bombesin antagonistic peptide containing the acid anino having the formula 1, 41, 2, 47, 47, 9, 10, 9, 10, -A-1 -A -A -A -A -A -A -

, 11, 12, 14, 14, 14, 14, 14, 14, 14, 14, 14, 14, 14, 14, 14, 2 A = Gin, Asn, Gly, Ala, Leu, He, Î ±, Î ± -aminobutyric acid, Vot, Val, Phe, pX-Phe (X = F, Cl, Br, OH, or C1-3), Trp, naphthylalanine or is eliminated; 35 1 5 10 15

Mod. 71 -10000 βχ. -89/07 20 25 30 61 «354 King: PlC / Bioraeasure-3ombesin CIPS. 5-7; 9; 10

Λ 3 = Arg, T> -Arg, Lys, D-Lys or is eliminated; 4A = Gn, Asn, Gly, Ala, Leu, Ile, Nle, amino (-aminobutyric acid Met, Val, Phe, pX-Fhe (X = α, Cl, Br, CH or cn), Trp, is eliminated, A is Gn, Asn, Gly, Ala, Leu, Ile, Nle, Alkococ-aminobutyric, Met, Val, Phe, D-Phe, pX-Zhe (X = F, Cl, Br, OII or CT ), Trp, N-ac-D-Ala, Leu, Ile, Nle, N-oxide, N-acyl, aminobutyric acid, Met, Val, Phe, pX-phe (X = F, Cl, Br, OH or Cl 3), Trp, pC-1a, naphthylamino or 4-substituted 7A = Gin, Asn, Gly, Ala, Leu , Ile, Nle, acid (-aminobutyric, Met, Val, Phe, D-Phe, pX-Phe (X = F, cl, Br, Oh or CH3), Trp, Lys, Ilis, or -naphthylianine; (X = F, Cl, Br, OII or CH3) TTp, Xaa, Xaa, Xaa, Xaa, Xaa, Xaa, or (Î ± -naphthyl anine; D or L; Î ±) = Gin, Asn, Gly, Ala, Leu, Ile, Nle, Aminobutyric acid, Met, Val, Phe, pX- Br, OII or CH 2), Trp, Thr, or (3-naphthylalanine; A11-Gly , Phe, D or L; A is Ilis, Phe, or p-X-Phe (X = F, Cl, Br, OII, C1-6), D or L; 13 3 A =. Gn, Gn, Gly, Ala, Leu, Ile, Nle, Acridoc-aminobutyric, Met, Val, Phe, p-X-Fhe (X = F, Cl, Br, Ch or CH3), Trp, or β-naphthylalanine; (X = F, Cl, Br, OII or CH3), Trp, Thr, Xa, Xa, Xa, Xa, or where R 2, R 3 and R 4 are independently selected from the group consisting of H, C 1-6 alkyl, C 1-6 alkyl, C 2-6 alkenyl, phenyl or naphthyl or phenylC 1-10 alkyl), or COOE 2 (wherein 4-C 1-10 alkenyl or phenyl-C 1-10 -alkyl) R 2 is attached to the N-terminal amino acid of the peptide which may be A 1 A 2, A 2, A 3, A 4, A 5, A 6 or A ', and also when one of R or R 2 is COE or CCOE ", the other has to be II, and when one of R 1 is O 2 or C 2 E 2, the other must be 15 35 1 5 10 15

Mod. 71 -10000 ex. · 89/07 20 30 61.864 Ref: PTC / 3iomeasure-Bombeain CIPS. 5-7; 9; 10

K, and also that when A is pGlu, R2 must have the portion of Clu which forms the imine ring in pGlu; and for each of the residues A, A, A, A, A and A, independently the carbon atom participating in the amide bond between that residue and the nitrogen atom of the alpha amino group of the adjacent amino acid residue may be a carbonyl carbon or may be reduced to a methylene carbon, provided that such a carbon atom at least has to be reduced to a methylene carbon, wherein the peptide also comprises A = Cys; A = Cys or an isomer D of any of the amino acids described for this position; 7A = Pglu, Cys, 1-methyl-Nis, or 3-methyl-Nis, or 3-methyl-N; 9A = Cy s; (Wherein X = F, Cl, Br, N0o, O11, or CTT), or the D-isomer of any one of Ala, Val, Gin, Asn, Leu, Ile, Mot, pX-Phe Trp, Cys, or fl-Nal; A 1 = methyl-His, or 3-methyl-His; 14 and where A can be eliminated. Preferably, the therapeutic peptide has the formula: Preferably, the amino acid sequence of the therapeutic peptides is as defined in claim 1, having the formulas described herein, homologous to at least the amino acid sequence of the naturally produced peptide; most preferably, this homology is at least 50 °. Non-peptide bonds in which the peptide bond is reduced are indicated herein by the symbol " " Y ". Antagonists of the invention may be used to treat diseases involving malignant or benign tissue proliferation, as all of cancer in which bombotin-related or PLG-related substances act on autocrine or paracrine analitic factors, for example gastrointestinal tract cancer, pancreatic cancer, colon cancer, lung cancer, particularly lung cancer, 61.S64 Ref; PTC / Dioneasure-Doeibesin CIPS. 5-7; 0; 10

1 5 10 15

Mod. 71-10000 e *. Or cancer of the seal, or to treat atherosclerosis, and gastrointestinal tissue disorders related to gastric and pancreatic secretions and motility; for example to cause suppression of amylase secretion, or for appetite control. In the generic formulas indicated above, when any of R-R or R is a lipophilic aromatic group, the activity in vivo may be of great duration, and release of the compounds of the invention towards the target tissue may be facilitated. The identifying group of an anionic acid is the Î ± -one or group of atoms, other than the carbon atom (-carbonyl, the nitrogen atom (-amino, or the Î ± atom) attached to the asymmetric carbon atom. examples, the alanine identifier group is ci-1, the valine identifier group is (CH) CH 2, the lysine identifier group is H 3 NT + (GI 0) 4 and the phenylalanine identifier group is CH 9, the identifying group of an acid] When the amino acid identifier group is not specified, this group may be (Î ±, Î ±, Î ±, Î ±, Î ±, Î ±, Other features and advantages of the present invention will become apparent from the following description of the preferred embodiments thereof and the claims. DESCRIPTION OF THE PREFERRED EMBODIMENTS Let us first give a brief description of the drawings. a curve chart of tumor growth for NCI-II69 xenografts. Figure 2 is a series of amino acid sequences of naturally produced peptides, peptides of which the peptides according to the present invention are analogous. Figure 3 is a graph depicting the blast effect of the bombesin analogue D-Phe DN (6-13) methyl ester on pancreatic amylase in bombesin-stimulated experiment. Let us now describe the structure, synthesis and use of CIPS. 5-7; 9; 10

1 5 10 15

Mod. 71 -10000ex. The preferred embodiments of the invention. Structure The peptides of the present invention all have a non-peptide bond at one of the positions indicated, at least, except the substituted stat-substituted analogs, such as Stat-DeLeu et al. Non-peptide bond means that the carbon atom that participates in the bond between two residues is reduced from a carbonyl carbon to a methylene carbon. The peptide binding reduction process is described by Coy et al., U.S. Patent Application Serial Nos. 879,348, issued to the same owner of the present application, incorporated herein by reference. Any of the amino acids at positions O, 1, 2 and 9 of the litorin antagonists can be elininated from the peptides, and the peptides remain active as antagonists. The peptides of the present invention may be provided in the form of pharmaceutically acceptable salts. Examples of preferred salts are those prepared with therapeutically acceptable organic acids, for example acetic, lactic, maleic, citric, malic, ascorbic, succinic, benzoic, salicylic, methanesulfonic, toluenesulphonic, or pamoic acid, as well as polymeric acids such as tannic acid or carboxymethylcellulose, and salts with organic acids such as hydrohalic acids, for example hydrochloric acid, sulfuric acid or phosphoric acid. Synthesis of Litorino Analogs 25 Synthesis of the litorino antagonist pC-lu-Pln-Trp-Ala-Val-Gly-His-Leu-CHO-Leu-MI0 will be described below. Other antagonists of bombesin, litorin, neuromedin or PLG may be made appropriate modifications of the following synthetic process. The first step is the preparation of the intermediate product pGlu-T / 1n-Trp-Ala-Val-Gly-1: 1 (benzyloxycarbonyl) -Leu-cH-Leu-benzhydrylamine as follows.

Benzhydrylamine resin (Vega Diochemicals, Inc. ((0.97 g, 0.5 mmol) as chloride ion is introduced into the reaction vessel of a Dockman 9903 peptide synthesizer programmed to perform the reaction cycle (a) methylene chloride: (b) 33% trifluoroacetic acid (TFA) was methylene chloride (2 times for 1 and 1)

Mod. 71-10000 ex. - 89/07 20 25 30 Οι · fiei: PTC / Biomeasure-Dombesin CIPS. 5-7; 9; 10

25 minutes each); (c) methylene chloride; (d) ethanol; (e) methylene chloride; and (f) 10% triethylamine in chloroform. The neutralized resin is stirred with alpha-t-butoxycarbonyl (Boc) -leucine and diisopropylcarbodiimide (1.5 molles each) in methylene chloride for 1 hour, and the resulting acidic amino resin is then cyclized through steps (a) to (f) in the above washing program. Aldehyde Boc-leticine (1.25 mmol), prepared by the process of Fehrentz e Castro, Synthesis, p. 676 (193) was dissolved in 5 ml of dry dimethylformamide (DIâ,ƒF), and the suspension was added to the suspension in the suspension, followed by the addition of 100 mg (2 mmol) of cyanoborohydride (100 mg, 2 mmol) (Sasaki and Coy, Peptides 8: 119-121 (19S7), Coy et al., After stirring for 1 hour, the resin mixture was negative for the ninhydrin reaction (1 minute), the The following amino acids (1.5 mmol) are then successively joined in the presence of diisopropylcarbodiimide (1.5 mmoles), and the resulting acidic amino resin is added by washing / unblocking steps ( a) to (f) in the above procedure: 3oc-Kis (benzyloxycarbonyl), Boc-Gly, Boc-Val, Boc-Ala, Boc-Trp, Boc-Gln (bound as a 6M excess of β- The resin described above (1.6 g, 0.5 mmol) is mixed with anisole (5 mL) and anhydrous hydrogen fluoride (35 mL), dried over anhydrous sodium sulfate ) a And stirred for 45 minutes. The excess hydrogen fluoride is evaporated rapidly under a stream of dry nitrogen, and free peptide is precipitated and washed with ether. The crude peptide is dissolved in a minimum volume of 2 M acetic acid and eluted on a (2.5 x 10) ram) column of Sephadex G-25 (Pharmacia Fine Chemicals, Inc.). Fractions containing a major component by uptake uv and thin layer chromatography (CCD) are then pooled, evaporated to a small volume and applied to a rcadecylsilane-silica (Y / hatman LRP- 1, mesh size 15-20 // m). The peptide is eluted with a linear gradient of 0-30% α-ketonitrile in 0.17% trifluoroacetic acid in water. The fractions are examined by CCD and by liquid chromatography of the major 191-1343

Ref: PTC / Biomeasure-Dombesin Cirs. 5-7; 9; (CHCl 3) and pooled to obtain maximum purity. Repeated lyophilization of the solution from water affords 60 mg of the product as a white, fuzzy powder. 5

The product is homogeneous when analyzed by CI.GR and CCD. Amino acid analysis of an acid hydrolyzate confirms the composition of the peptide. The presence of the I.eu.Leu-Leu-4 binding is demonstrated by the rapid atom bombardment mass spectra. p-Al-Val-Gly-Ile-3-Î ± -Î ± -Î ± -Î ± -β-Al Î ± -L-Val-Gl other peptides are pr analogue color; modification and / CTI, J2I | Leu-Mi, or epared in similar yields appropriate from the procedure above. The solid phase synthesis of D-Fhe in a manner 15

Mod. 71 · 10000 ex. -89/07 20

Leu ^ Y | C: D-β-litorin, D-Phe-Cln-Trp, Ala-Val-Gly-lys-Leu, The synthesis was carried out in the following manner: Boc-D-Fn-Gln-Trp, Ala-Val-Gly-IIis (tosyl Benzhydrylamine-polystyrene resin (Advanced Clinken, Inc.) (1.25 g, 0.5 mmol) was introduced as the chloride ion in the vessel (a) to (f) above. The neutralized resin is maintained under stirring with Brc-D-phenylalanine and diisopropylcarbodiimide ( 1.5 mmol each) in methylene chloride for 1 hour and the resulting acidic amino resin is then cyclized through steps (a) to (g) in the above washing program, the Boc group is then separated by treatment with Dissolve Doc-leucine aldehyde (1.25 mmoles), prepared by the Fehrentz and Castro process (1), in 5 ml of dry THF and add to the resin salt suspension, sec The addition of 100 mg (2 mmol) of sodium cyanoborohydride (2.3) was added. After stirring for 1 hour, the resin mixture is observed and it is found that the reaction is negative for the nihidrin reaction (1 minute), which indicates complete derivatization of the free amino group.

The following amino acids (1.5 mmol) are then joined successively by the same method; 5 10 15

Mod. 71 -10000 ex. - 89/07 20 30

Ol.àdA Ref: PTC / Biomeasure-Dcmbesin CIP3. 5-7; 9; 10

Boc-Ala, Boc-Trp, Boc-Gln (linked in the presence of 1 equivalent hydroxybenzotriazole), Boc-T) -Phe (bound in the presence of 1 equivalent hydroxybenzotriazole ). After drying, the peptide resin weighed 1.93 g. The rosin (1.93 g., 0.5 mmole) is mixed with anisole (5 mL) and anhydrous hydrogen fluoride (35 mL) at 0Â ° C and stirred for 45 minutes. Evaporate excess hydrogen fluoride under a stream of dry nitrogen, precipitate out free peptide and wash with ether. The crude peptide is dissolved in a minimum volume of 2% acetic acid and eluted on a Sepha-dex G-25 column (2.5 x ICO mm). Fractions containing a major component are then pooled by UV absorption and thin layer chromatography, evaporated to a small volume and applied to a column (2.5 x 50 cm) of octadecylsilyl in Vydac (10-15 // '.'). This is eluted with a linear gradient of acetonitrile 15-45% in trifluoroacetic acid 0.10) in water. The fractions are examined by high performance analytical liquid chromatography and pooled to obtain the highest purity. Repeated lyophilization of the solution from water affords 120 mg of the product as a fluffy white foot. Other peptides, for example D-β-Cl-Phe-Gln-Trp-Ala-Val-Gly-IIis-Leu -B-Phe-tH1, can be synthesized by essentially the same procedure. The solid phase of the synthesis of D-Phe-Gln-Trp-Ala-Val-G1 and-Ili s-Leu | CII-NII | -D-Pho-NII2 was performed as follows: Boc-B-Phe-Gln-Trp-Ala-Val-Cly-IIis (tosyl) -Leu Cí0 \ n | -D-Phe-benzhydrylamine was synthesized first. Benzhydrylamine-polystyrene resin (Advanced Cher Tech, Inc.) (1.25 g, 0.5 m-Oles) is introduced as chloride ion into the reaction vessel of an Advanced Cliem Tech ACT 200 peptide synthesizer programmed to effect the following reaction cycle: (a) methylmercury; (b) 33% trifluoroacetic acid in methylene chloride (7 times for 7 minutes each); (c) methylene chloride; (d) 35 1 5 10 15

Mod. 71 10000 ex. - 89/07 20 25 30

- > 1. SC-i Ref i PTC / Rio ::; easare-Bombeoin 3IP5. 5-7; 9; 1G

ethnol; The resin neutralized was stirred for 1 hour at 0 ° C and the resulting amino acid resin was stirred for 1 hour and the resulting amino acid resin was evaporated to dryness. The Boc group is then separated by treatment with ΪΓΛ Boc-leucine aldehyde (1.25 mmoles), prepared by the process Fehrentz and Castro (1) were dissolved in 5 ml of dry DMF and partitioned between the resin TFA salt suspension, and the addition of sodium cyanoborohydride (2.3 ml), 100 mg (2 ml). After stirring for 1 hour, the resin mixture is observed to be negative for the ninhydrin reaction (1 minute), which indicates complete derivatization of the free amino group. The following amino acids are cleaved successively by the same procedure: 3oc-1β- (benzyloxycarbonyl), 3β-Gly, 3β-Val, Boc-Ala, 3β-Trp, B (attached in the presence of 1 equivalent hydroxy-benzotriazole), Boc-D-Phe (bound in the presence of 1 equivalent hydroxybenzotriazole). After drying, the peptide resin weighed 1.93 g. The resin (1.93 g, 0.5 mmol) is mixed with ani-sol (5 ml) and anhydrous hydrogen fluoride (35 ml) at 0 ° C and kept stirring for 45 minutes. The excess hydrogen fluoride is rapidly evaporated under a stream of dry nitrogen, precipitated from free peptide and washed with ether. The crude peptide is dissolved in a minimum volume of 2 M acetic acid and eluted on a column (2.5 x 100 mm) of Se-phadex G-25. Fractions containing a major component by uv uptake and thin layer chromatography are then pooled, evaporated to a small volume and applied to a column (2.5 x 50 cm) octadecylsilene Vydac (10-10 ÂμM ). This is eluted with a linear gradient of 15-45% acetonitrile in trifluoroacetic acid in water. The fractions were examined by thin layer chromatography and high performance liquid chromatography and roughed for maximum purity. Repeated lyophilization of the solution from water affords 120 mg of the product as a white, fuzzy powder. It is observed that the product is homogeneous by means of CLCR and CCD. Amino acid analysis of an acid hydrolyzate confirms the composition of the otapeptide. The presence of the peptide linkage LeuS ^ ICHgNIf is des-22- 1 5 10 15

Mod. 71-10000 ex. - 89/07 20 25 61.864 Ref: PTC / niomeasure-Borabesin CIPS. 5-7; 9; 10

shown by mass spectrometry of fast atom bombardment. The solid phase of the synthesis of D-Fhe-Gln-Trp-Ala-Val-Gly-His-Leu-NH was carried out in the following manner. Step (1): Tenzhydrylamine-poly-styrene resin (Advanced Chem Tech, Inc. (0.62 g, 0.25 mmol) is introduced as the chloride ion into the reaction vessel of an ACT 200 peptide synthesizer programmed to provide the following reaction cycle: (a) methylene chloride: (b) 33% trifluoroacetic acid in methylene chloride (2 times for 1 and 25 minutes each): methylene chloride (d) ethanol; ) The neutralized resin is stirred with 3β-leucine and diisopropylcarbodiimide (1.5 moles each) in methylene chloride for 1 hour and the resulting acidic resin is then added The following amino acids (1.5 mols) are then successively joined by the same procedure: Boc-IIis (benzyloxycarbonyl), Boc-Gly, Hoc -Val, Boc-Ala, Boc-Trp, Boc-Gln (attached as a 6æl excess of p-phenyl ester, and pGlu (bound in the presence of hydroxybenzotriazole). After drying, the peptide resin weighed 0.92 g. Step (2): The resin (0.92 g) is then mixed with anisole (5 ml), dithiothreitol (200 mg) and anhydrous hydrogen fluoride (35 ml) at 0 ° C and kept under stirring for 45 minutes. The excess hydrogen fluoride is rapidly evaporated under a stream of dry azodc, precipitated from the free peptide and washed with ether. The crude peptide is dissolved in a minimum volume of acetic acid. and eluted on a column (2.5 x 100 cm) of Sephadex G-25. Fractions containing a major component are then pooled by uv absorption and thin layer chromatography, evaporated to a small volume and applied to a column (2.5 x 50 cc) octadecylsilane Vydac ( 10-15 microir). The column is eluted with a linear gradient of 0-30% acetonitrile in 0.1% trifluoroacetic acid. The fractions are examined by thin layer chromatography and pooled to obtain the highest purity. Repeated lyophilization of the solution from water affords a fleshy, biodegradable powder; it is observed that this product is homogeneous by means of CI-HE and CC3. 30 1 5 10 15

Mod. 71 -10000 ex. - 89/07 20 25 30

51 · 354 Ref; ΓTC / í e e e e e e e e e e e e e e e e. 5-7; 9; 1C

Amino acid analysis of an acid hydrolyzate confirms the composition of the peptide. The reaction was cooled to 0 ° C. The reaction mixture was cooled to 0 ° C. The reaction mixture was cooled to 0 ° C. The reaction was cooled to -78 ° C. (1), and 0.92 g in step (2)), Synthesis of N'-acetyl-D-Phe-C-1-Trp-Val-01-Ile-Leu (0.25 g, 0.25 mmol) of benzhydrylamine resin in step (1), and mixing 0.92 g of the resin with anisole in step (2), except that the final Ooc group was separated and the resin was acetylated with acetic anhydride in methylene chloride. δ 9 synthesis of Sta-Des-Met Litorina A statin residue, AIIPH or ACHPA, instead of any two amino acids of the peptide, may be substituted for the peptide. , S deo contains only peptide bonds. For example, this compound was prepared analogously, starting by attaching the statin to the resin and then proceeding with the addition of Roc-IIis (benzylcarbonyl). Synthesis of statin or Boc-statin can be done according to the procedure of Rich et al., 1978, J. Org. Chem., 43; 3624; and Rich et al., 1980, J. lled. Chem. 23; 27, and AHPPA and ACIIPA can be prepared by synthesis according to the procedure of Ilui et al., 1987, J. F. Chem. 30: 1287; Schu-da et al., 1938, J. Org. Cliem. 53: 873; and Rich et al., 1988, J. Org. Chem. 53: 869. The solid phase synthesis of peptide 3β-26120, pGlu-Gln-Trp-Ala-Val-Cly-IIis-Sta-NII, was effected by the application of the following procedures, in which alpha-t-butoxycarbonyl (prepared by the process of Rich et al., J. Chem., 197, 43, 3624) with netilbenzhydrylamine-polystyrene resin. After acetylation, the pGlu-Gln-Trp-Ala-Val-Gly-His (benzyloxycarbonyl) -Sta-methylhenzhydrylamine resin intermediate was prepared. The synthetic procedure applied for this preparation is described in detail below: 1. Incorporation of alpha-t-butoxycarbonyl statin onto methylbenzhydrylamine resin. Methylbenzhydrylamine resin (Vega 24- 1 5 10 15

Mod. 71-10000 ex.-89/07 20 25 61.S04 Ref: PTO / Biomoa3ure-Bombesin CIPS. 5-7; 9; 10 3 ^ 1990 10

Biochemicals, Inc.) (1.0 g, 0.73 mmol) as the chloride ion in the reaction vessel of a Vega 250C Coupler peptide synthesizer. The synthesizer was programmed to carry out the following reactions: (a) methylene chloride; (b) 10% triethylamine in chloroform; (c) ne-tylene chloride; and (d) dimethylformamide. The neutralized resin is mixed for 1 hour with the active ester formed above from alpha-t-butoxycarbonyl statine (1.46 mols), diisopropylcarbodiimide (2 mmoles), and hydroxybenzotriazole hydrate (1.46 mmol) in dimethylformamide at 0 ° C for 1 hour. The resulting acidic amino resin is washed in the synthesizer with dimethylformamide, and then with methylene chloride. The resin blend at this point had, when subjected to the Iieiser ninhydrin test (5 minutes), an 84% level of statin incorporation into the resin. Acetylation was carried out by mixing the amino acid resin for 15 minutes with N-acetylimidazole (5 mols) in methylene chloride. The derivation for the 94-99% level of the resin amino groups of the resin was indicated by Kaiser's ninhydrin tost (5 minutes). The boc-statin-resin is then washed with methylene chloride. 2. Amino Acid Remaining Amino Acids. The peptidic synthesizer is programmed to carry out the following cycle of reactions: (a) methylene chloride; (b) 33% trifluoroacetic acid (TFA) in methylene chloride (2 times for 5 and 25 minutes each); (c) methylene chloride; (d) isopropyl alcohol; (4 mmoles) plus diisopropyl carbodiimide (4 mmoles) plus diisopropylcarbodiimide (4 mmoles) plus hydrate of hydroxybenzotriazole (1.47 or 0.73 mols) and the peptide resin is washed in the synthesizer with dimethylformamide, and then methinene chloride, and then quenched by washing and unblocking steps (a) through (f) in the procedure described above.

Boc-Li (benzyloxycarbonyl) (attached in the presence of 2 equivalents hydroxybenzotriazole); Boc-Cly; Boc-Val; Boc-Ala and -25- 35 1 5 10 15

Mod. 71-10000 ex. -89/07 20 25 30 G1.S64 Re f: PTC / Biorne ns ure-Bombe si n C1PS. 5-7; 9; 10

Boc-Trp (bound as the pre-formed hydroxybenzotriazole active esters prepared by reaction at 0Â ° C for one hour with equivalent hydroxybenzotriazole hydrate); Do-Gln and pGlu (also bound as the pre-formed active esters of hydroxy benzotriazole, prepared by reaction at 0 ° C for 1 hour with 1 equivalent of hydroxybenzotriazole hydrate). The completed peptide resin is then washed with methanol and air dried. The peptide resin described above (0.60 g, 0.73 mmol) is mixed with anisole (2.5 ml), dithioerythreitol (50 mg) and anhydrous hydrogen fluoride (BC ml) at 0 ° C for 1 hour . The excess of hydrogen fluoride is rapidly evaporated under a stream of dry nitrogen, precipitating the free peptide and washing with ether. The crude peptide is dissolved in 100 ml of acetic acid, and the solution is then evaporated under reduced pressure. The crude peptide is dissolved in a minimum volume of 1/1 methanol / water and triturated with 10 volumes of ethyl acetate. The triturated peptide was applied to a column (9.4 mm ID x 50 cm) of octadecylsilane-silica (Whatman Partisil 10 0DS - 2 M p). The peptide is eluted with a linear gradient of 20-80% of 20/80 0.1% trifluoroacetic acid in water. The fractions are examined by means of CCD and high performance analytical liquid chromatography (CLGR) and pooled to obtain the highest purity. Lyophilization of the solution from water affords 77 mg of the product as a white, fuzzy powder. Other compounds, including D-Cpa, α-leu, des · 9 Met Litorin, may be prepared as described above and assayed for efficacy as agonists or antagonists in the following test program. Phase 1 - 3T3 Peptide Uptake Assay Estimulated I Hlcom Thymidine Cell Culture. Cultures of 3T3 Swiss cell cultures are grown in Dulbecco's Modified Eagles (I.IEMD) supplemented with 10% fetal calf serum in a humidified atmosphere of 10% → 90% air at 37 ° C. For experimental use, the cells are seeded in trays with groups of 24 wells and used four days after the change of medium. Cells are detained in the G1 / G0 phase of the 26-b + cycle. iíb4 Ref: PTC / Biomeasure-Bombesin CIPS. 5-7; 9; 10

1 5 10 15

Mod. 71-10000 ex. Cells by changing the serum free MEVD 24 hours prior to the thymidine uptake assay. Synthesis DNA. The cells are washed twice with 1 ml aliquots of DMEM (chloroform), then incubated with DMF (chloroform), 0.5 Methyl H-thymidine (20 g / mmol, New England Nuclear), bombesin (3nM), and initially four concentrations of the test compounds (1, 10, 100, 1000 nM) in a final volume of 1.0 ml. After 23 hours at 37 ° C, methyl thymidine was incorporated into insoluble acid groups, and then this incorporation is assayed as described below. Cells are washed twice with ice-cold 0.9 Na NaCl (1 ml aliquots), and acidic radioactivity is separated by a 30 minute (42C) incubation with trichloroacetic acid (TCA). The cultures are then washed once (1 ml) with ethanol at 95 " are solubilised by means of a 30 minute incubation (1 ml) with 0.1 N NaOH. The solubilised material is transferred to vials containing 10 ml of ScintA (Packard), and the radioactivity is determined by means of liquid scintillation spectrometry. Phase 2 - Small Cell Carcinoma (5C1.C) Capture Assay with Bombesin ι Η I Thymidine. CCelular Culture. cultures of the human cell from the carcinoma cell line. (NCI = HB9) (obtained from the American Type Culture Association) are maintained in P.FVI 1340 medium supplemented with fetal calf serum at 10Â ° C in 10Â ° C / 90Â ° C air at 37Â ° C. Twenty-four before one assay, M cells are washed with serum-free medium and seeded in trays with 24-well groups. DNA Synthesis Experience. Good amine (1nM), C, 5 / (20 Ci / mmoles, New England Nuclear), and four concentrations of the test compounds (1, 10, 100, 1000 nM) were added to the cultures to give a final volume of 0.5 ml. After incubation at 37 ° C, the cells are pooled on CF / B glass fiber filters, and the DNA is precipitated with ice cold TCA. Λ incorporation of thymidine | 3II | in DNA acid insoluble fractions is determined by means of liquid scintillation spectrometry. Phase 3 - Peptide Induced Pancreatitis Male Sprague-Dawley rats (250 g >

Mod. 71-10000 ex. - 89/07 20 25 30 æl. (3-1 Rof: PTC / Biomeaaure-Bombesin CIPS, 5-7; 9; 10

for these experiences. The test compound, or 0.9% NaCl, is given a s.c. 15 minutes prior to the bombesin injection. Bombesin injections are given s.c. at a dose of 10 μg / kg, and blood samples are obtained at 1 h 30 m, 3 h and G hours. Plasma amylase concentrations are determined by the Amylase Vantrak assay. Phase 4 In Vitro Inhibition of Gastrin Releasing Peptide Binding 125, II a 3ombesin Receptors Membranes from various tissues (rat brain, rat pancreas, anterior pituitary rate, SCLC cells, 3T3) and homogenization in 50 mM TrisHCl containing 0.1% bovine serum albumin and 0.1 mg / ml bacitracin, followed by two centrifugations (39,000 xg × 15 min, 42 ° C) with an intentional resuspension in buffer freshly prepared. For experimentation, 125 aliquots (0.1 ml) are incubated with 0.5 nM (2000 cc / nmoles, Amersham Corp.) and various concentrations of the test compounds in a final volume of 0.5 ml. After a 30 minute incubation at 4 ° C, the ligation reaction is complete with rapid filtration through GF / C filters which were previously soaked in 0.3% aqueous polyethyleneimine to reduce the level of nonspecific binding. Filters and tubes are taken three times with aliquots of 4 ml of ice-cold buffer, and the radioactivity trapped in the films is counted by means of a spectrophotometer. Λ specific binding of-fine-as connection | PLG minus that binding in the presence of 100æM bombesin. Phase 5 - Inhibition of Gastrin Release Stens from anesthetized rats are sprinkled with saturated sodium chloride solution over periods of 15 minutes via pyloric cannulation, while the experimental peptide is infused through the femoral vein for periods between 0 c 150 minutes. Stage 6 - In Vivo Antitumor Activity MCI-II69 small cell lung carcinoma cells were transplanted from culture In vitro, by implantation in each animal of the equivalent of 5 75 cm 2 confluent tissue culture flasks on the right flank. The culture of NCI-II69 cells is added in vitro as a suspension of cell pellets. Pcr-23- 10 15

Mod. 71-10000 ex. -89 / 07 20 25 30 35 J. · o ·:; - * 'ef: PTC / bi orneasure-Borabesin n CIP3. 5-7; 9; Thus, no attempt was made to disaggregate cell clusters by physical or chemical means. The tumor size was calculated as the mean of two diameters, i.e. (length to width / 2) mm. Results of cSc Hssa Psychotic Experiments

Various putative or pGG analogs, each containing a peptide bond or a sstatin, ANFRA or ACURA residue, may be synthesized and tested in one or more of the experiments. The results of the Phase I and Phase 2 trials are shown in Table 1. Table 1 gives the following formulas for 03 analogs: 12: non-peptide yogas and in vitro binding inhibition results PLG receptors3 from bombesin fibroblast 373, and uptake of bombesin-stimulated β-adenine by cultured 3T3 cells (Data on 3T3 PLG receptors and thymidine uptake are expressed as IC50 (nM). also indicates results for analogs containing non-peptide binding of another naturally occurring peptide, Neuromodin C, the seven C-terminal amino acids being similar to bombesin and PLG aces (Table I, " Litosin " refers to a residue 9 analogue peptide or its derivative whereas " Neuromodin C " refers to an analogue of residue 10 or its derivative.). In Table I the position of the non-peptide bond is indicated by the position of the symbol (CH 2 NH 3), i.e. CH 2 NHH is always represented by the amino acid which in that peptide is attached to the N-terminal amino acid When no amino acid is specified under " structure ", the non-peptide bond binds the two peptides represented by the numbers indicated as post-written. In Table I, it can be seen that a placement preferably of the non-peptide binding in litorin analogs is in the A-A position, two of the most active analogues (indicated by a small variant IC 50 receptor) are 3IM-26100 (Fhe In addition, as shown in Table X-8118 I, BIN-26113 (D-Phe, Leu ') and the BIM-2G101 (Phevy-CH3NH-Leu9) CII-NH Leu) and BIM-26114 (D-Nal, Leu2, Q3, CH3 NH Leu) are active at the 3T3 PLG receptor binding and in the experiments of In most striking way, BIM-26136 (D-Nal-29-51015

Mod. 71-10000 ex. -89 / 07 20 25 (51,364 Bei: PTC / Biomeasure-Borabesin CIPS, 5-7: 9: 10-9: 9: 89, Leu γ fCH2NII (Fhe), which contains amino and terminal carboxylic aromatic residues, which may form a hydrophobic interaction, is the most potent analogue P-3-leucine substitutes residues A and litorin, the resulting analogues BIi-26120 and 3IM-231S2 are also potent antagonists. Table I also shows Neuromedicillin analogs containing a peptide bond between β-A residues, for example 3DI-26092, 26095, 26106 and 26107, are antagonists when tested at the PLG receptor 373 and in the uptake experiments of thymidine. Table I also gives negative results for Neuropedin C analogues, for example BIM-26108. Thus, the non-peptide binding lines indicated herein should be used in conjunction with the routine experiments described above to choose usable antagonists. Bombesin and Bombesin analogues have been shown to inhibit the effect of interleukin-2 (IL-2) (Fink et al., 1983, Klin Vossenschr. 66, Suppl 13, 273). Since 1L-2 causes proliferation of T lymphocytes, it is possible that litorin antagonists may prevent the inhibitory effect of Bombesin or its analogs on UL-2. IL-2-stimulated lymphocytes can cause lysis in small cell lung carcinoma cells in vitro. Although Dombasin antagonists have an antiproliferative effect on neoplastic tissues, they may also facilitate the proliferation of lymphocytes which have lytic activity for small cell lung carcinoma. These observations led us to evaluate the effect of BIJ-26100 on the in vivo culture of the SCLC tumor cell line described in Step 6. Twenty athymic bare females, 5 to 6 weeks old, were implanted on day 0 with human SCLC NCI-H69, had been individually identified, and then were randomized to the following control vehicle test groups: Ref: PTC / Biomeasure-Bombesin CIPS. 5-7; 9; 10

1 10 15

Nfo Group. 1

Treatment No. of Animals Control treated with vehicle sodium chloride saturated solution 0.2ml, s.c. inf., B.i.d. QDL-28 10 2 3IM-26100: 50 g / inj, s.c., b.i.d., QDL-2853 31: .1-26100: 50 g / inj. s.c., inf., b.i.d., QDL-285 (s.c. = subcutaneously; in. = injected; b.i.d. = twice daily; QD1-28 = daily treatment on days 1-28).

Mod. 71 -10000 βχ. · 89/07 20 25 The growth of xenograft NCI-II69 and the tumor growth inhibitory activity of the bombesin antagonist BIú-26100 (pGlu-Gln-Trp-Ala-Val-Gly-Ile-PheY · CK · NH Leu-NTI4) are represented by tumor growth curves in Figure 1, and the relative tumor dimensions in Table II. Administration of 3β-261CC infusion to s.c. around the tumor inhibited significant nerve growth. The efficacy of the antitumor antisense of BIJT-2610C is evident in view of large Î »1-1169 inoculum tumor cells (i.e., the equilibrating equivalent of 5 75 cm cell culture flasks " nim.al) and the agglomerated state of the cells. In confluent flasks, XCI-II63 α-glues are microscopically visible, and, in conglomerate, resemble a metastatic tunor colony. Many of these tumor colonies were implanted by animal. The dose of B1M-261CO was chosen to be arbitrarily based on the availability of compound and not optimal. Larger doses of RIM-261CC, as indicated by body weight gain (less weight of tvnor) obtained during treatment (Table III), may be given. This appears to indicate that BIM-26100 does not have any local or systemic toxicity and is therapeutically usable as an anti-growth factor with antitumor effects. Figure 3 depicts the effect of the bombesin antagonist D-β-Cl-Phe-Gln-Trp-Ala-Val-Gly-IIis-Eeu® CHÎ'-XIII on the secretion of bombesin-stimulated amylase in the rat. The results show that this analogue is a potent antagonist; 5 nV of the analogue may -31-35 61,834

PTT / 71iomeasure - BoEibesin CIP3. 5-7; 9; Inhibition of stimulated amylase secretion by 0.5 nM bombesin for 150 minutes after injection of the pill.

Use

The peptides according to the present invention may be administered to a mammal, in particular a human, in one of the traditional (e.g. orally, parenterally, transdermally or transmucosally) in a sustained release preparation using a biodegradable bicompatible polymer, by spot release (for example, in the case of anticancer bombesin to the lungs) using micelles, gels and liposomes.

The bombesin antagonists of the invention are suitable for the treatment of all forms of cancer in which bombesin-related substances will act as autologous or paraacrine agents, in particular small cell lung carcinoma. Peptides may also be used for the inhibition of gastric acid secretion and gastrointestinal tract motility disorders, sitntomic relief and / or treatment of exo-crinic pancreatic adenocarcinoma, and restoration of cachectic patient appetite. The peptides may be administered to a himanian patient at a dose of 0.5 μg / g / day. For some forms of cancer, for example small cell lung carcinoma, the preferred dose for curative treatment is 250 mg / patient / day. -32- 10 15

Mod. 71 -10000 ex. -89/07 20 25 30 ul.So4 Ref: P7C / 3ioneasure-Bombesin CIPS. 5-7; 9; 10

TABLE 1 code Structure BI'-26092 Gly-Asn-Hia-Trp-Ala-Val-Gly-Ilis-Leu-Leu-Ni-3I2-26095 pGlu-Gln-Trp-α-Val-D-Ala - IIis-LeuM > L-Glu-Gln-Trp-Ala-Val-Gly-His-PhG-C02-ΊΠ Leu-NII Litorin BIM-26101 pGlu-Gln-Trp-Ala-Val Glu-Ala-Val-D-Ala-Kis-Leu-CH 2 CH 3 -L-Ala-Asn-His-Trp-Ala-Val- -26106 D-Ala-IIis-Trp-Ala-Val D-Ala-IIis-Leu-C-Cl-N-O-Tet-MH0 Neuromedine C 3 -IM-26107 D-Phe-His-Trp-Ala-Val-Gly - N-Ac-D-Ala-IIis-Trp-Ala-Val-Gly-IIis-Leu-CIII-NFl3 Leu-MI, Neuronedin C 3124-26113 O-Pho-Cln-Trp-Ala-Val-Gly-Alis-Lys-Li-MeIII-Li, 3124-26114 n-Gal-CIn-Trp-Ala-Val -Gly-Ilis-Lev.

PLG 373 IC 50 receptor (n, '.!)

C hymidine uptake IC 50 (nM) 242 466 2623 1209 23 26

11S 107 401 199 841 296 107 354 154

The COO 35 < sX 10 15

Mod. 71-10000 ex. -89/07 20 25 30

Hof: PTC / 3iomeasure- Bombesin CIPS. 5-7; 0; 10 TABLE I (Continued)

3IM-26123

3IM-2613S 3IM-261S2

Structure of pClu-Gly-Trp-Ala-Val-Gly-Ile-Leu: Litorin D-Nal-Gln-Trp-Ala- Val-Gly-IIis-Leu and CCII2NUH Fhe-N7I-Liriamine DC p-C-1 n-Trp-Ala-Val and Glu-L-

PLG Receptor 3T3 IC'0 (nV)

Capture of tinine ICGCCr,;) 150 155 5.9 28.6 1.40.88 3.3 4.77 -34-35. »Hef: PTC / 3ioneasure-Bonbesin CIPS.5-7; 9; 10%

Mod. 71-10000 ex. - 89/07 10 15 20 25 30 z &gt; or a pharmaceutically acceptable salt thereof, such as a pharmaceutically acceptable salt thereof. -CO-C-Cl &gt; co + 1 + 1 + 1 + 1 + 1 + C H o C o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o n fO < C o-C o alkyl; (A) a compound of the formula: M H O H r H ca v; &lt; RTI ID = 0.0 &gt; RH &lt; / RTI &gt; &lt; / RTI &gt; + 1 + 1> 0O rH * d + 1 H IO r '. • • • • • • • • • • • • • • • • • • • • • • • • • • • • • P 00 • • H T3 • CM • HH »| λ • Ci Ο P Ό Q • .Ω d G * O • t ~ 4 P • O r- * d • 71 • • 6 * <T3 01 H * P · o rH • H ~ 5 • C3 O · V "> rH ^ 0 • Η gH - X • H o ϋ · ta ta WP νΉ · Ο« Η Π 5 3 Ο ΙΩ P Ό «Ο Ο Ο Ο Ο Ο Ο Ο Ο Ο Ο Ο Ο Ο Ο Ο 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 Ο Ο Ο Ο Ο Ο Ο Ο Ο Ο Ο Ο Ο Ο Ο Ο Ο Ο Ο Ξ Ξ Ξ 3 c .: co 00 r- * «k C0 Ο O a u u (((((((((((Φ Φ Φ Φ Φ Φ Φ Φ Φ Φ Φ Φ Φ Φ Φ Φ Φ Φ Φ Φ v / v at 0 ° C). 5 o o o o c o o Sn Sn Sn Sn Sn Sn Sn Sn Sn Sn Sn Sn Sn Sn Sn Sn Sn Sn Sn Sn Sn Sn Sn Sn Sn Sn Sn Sn Sn Sn.

The B O ϋ Cl O • c C3 O • H * o c • H U O * o tf Q

c H

3 H-NMR (DMSO-d 6)?

The

a or H

35 15 10 15

r-. The 20- 30 30 U1.0¾¾Ref: PTC / 13iomeasure-Bombeain CIPS. 5-7; 9; 10 ka

• • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • 0 &lt; 3 &gt; 01 $ c: o 1 o o o o o o o o o o o o o o o o o o

&lt; CJ to 'W οι H I w

C38 w R «or coI ►H U HH8 H § < t-i and «w co-3

nI K CJco and r '* 0 ω c κ fc. a β 1h o au o u 0 01 φ Ph

C H H H H H H H H H H H H

β o O n d r a a · k · d d. (0 c-0 rH H rH u O o CC m Cl 00 * Cl • • rH • O • rH c * • • * 00 • • k Cl • • • • • • • • • • • • • • • • • • • 0 4 • • • • • • • • • • • • • • • • • • • • • • • • • • - •? H H H r H • H 3 • X X o • H b b b D SH 0) c • rH * H,> O-O-O-O-O-O-O-O-O-O-O-O-O-O-O-O-O-O-O-O- 3 u »» »» »» »» »» »» »» »» »» »» »» »» »» »» »» »». 03 Ο Ο ϋ Η- &gt; a • Η κ rH ca © C. • Η dd Cl d Ό »01 ε l- * k 1η 0 0 Ο a ο ϋ 3 ΙΗ ta β tfl Β Ο β (11 ε ο ο φ a C ca 0) L0 0 Ό • α 0) d β Ε Ο 0 • ο Η 0 ΝΗ 0 «Η β U ca 1η Η-> C • Ρ β d £ ο * < 3 ο Η-> β 0 Ε Ε β ε β tr 01 ε Ο Η φ • W β ε ca • k ιΗ Φ tí r · a U 0 ε • Η <rs ο β rH Ε Μ X Φ α Cl ο Uβ ca Η-> ; Ρ Φ Η Η Η Ό Ό Ό Ό Ό Ό Ό Ό Ό Ό Ό Ό Ό C1 C1 C1 C1 C1 C1 C1 C1 C1 U U C1 U U U U U U U U U Ρ Ρ Ρ Ρ Ρ Ρ Ρ Ρ Ρ Ρ Ρ Ρ Ρ Ρ Ρ Ρ Ρ Ρ Ρ Ρ Ρ Ρ Ο Ο Η Η © © © © U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U ψ-t 1η C1 Ρ β α Μ Ο 0 τί β η Η δ ca 3 α 0 ε ca 1η η Φ Ό Ο Λ ΊΗ

36 '35 1 * ο5 * -1 ftef: FTC / I3iomoa3urQ-Sor.ibesin CII-3. 5-7; 9; 10

10 15

Mod. 71-10000 ex. - 89/07 20 30

Li torjr.a AI Λ2 A3 Λ4 Λ5 Λ6 Λ7 ΛΒ A9 pGlu-Gxn i-Trp-Al; a-Yal -Cl; and-β-β-β-β-β- O-Ser-Is- • Trp-Ala, and n-V-Cly-lys-Leu -; (a) Amino Acid Amino Acid Amino Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid AI A2 A3 A4 A5 A6 A7% CO < 1-Tris-Ala-Val-Gly-Hys-Leu-1, and FIG. 2. 35

Claims (5)

1 10 15 Mod. 71-10000 ex. - 89/07 20 25 30 61 · OG4 Ile: PTC / Niomeaurein nc: nbesin CIPS. 5-7; 9; 10-i! INDICATIONS- 15. A process for the preparation of a linear peptide which is a naturally occurring biologically active amphibian amphibian analogue or mammary PLC which is an active site or a binding site responsible for the attachment of said peptide to a receptor in the cleavage of a peptide bond at said active site of naturally produced bombosin or PLC for in vivo biological activity is unnecessary, characterized in that said analog has one of the following modifications: (a) elimination of a residue within said locus and b) a substitution of one or two residues within said active site with a synthetic acid, said analyte being capable of binding to said receptor, in a manner that said analog is capable of acting as an inhibitor concurrent with said naturally occurring peptide by means of a binding According to one of the foregoing modifications, the biological activity of said naturally occurring bombesin or Π-Π, 2-Π, is reduced. The composition of claim 1, wherein said active site comprises an amino acid at least at the carboxy terminus to the peptide medium, said linear peptide amino acid including at its carboxy terminus. A process according to claim 1, wherein said active site includes an amino acid at least in the amino terminus of the peptide, said linear peptide including said amino acid in its amino terminal moiety. A process according to the preceding claims, comprising from seven to nine amino acid residues, inclusive, characterized in that the peptide prepared is an analogue of one of the following naturally occurring peptides terminating at the carboxy-terminus with a residue, Tet (a ) ldtorin; neuromedin; (c) 03 ten amino acids of the carboxy terminal zone of mammary gastrin releasing peptide; and (d) the ten amino acids of the carboxy-terminal zone of amphibian bombesin, said therapeutic peptide having the formula: -33 · 10 15 Mod. 71-10000 ex. - 89/07 20 25 30 Cl.864 Ref; FTC / diomeasure-Bombesin CIP3. 5-7; 9; 10 Gly, Nle, d-α-inobutyric acid, or the D isomer of any of Claims 1 to 4, wherein R 1, R 2, R 3, (Where X = F, Cl, Br, NC0, C1-6 or CII3), Trp, · Cys, or?, Nal, or is eliminated; the D- or L-isomer of any one of pGlu, Me or acylamino-aminobutyric, or the D-isomer of any of Ala, Val, Cln, Asn, Leu, Ile, Met, pX-phe wherein X = F, Cl, Ur, N ° 2 or -Mal, or is eliminated; pGlu, Gly, Ala, Val, Gin, Asn, Ile, Ile F, Cl, 3r, NO 2, OH, II or CII 2) , Trp, Cys or 3-methyl-IIis; 4A-Ala, Val, Gin, Asn, Gly, Leu, Ile, Me, Amino-butyric acid, Met, pX-Phe (wherein X = F, Cl, Br, NO ", CK, II or CH) , Trp, Cys, or (3-Mor; 5 Λ Cln, Asn, Gly, Ala, Leu, Ile, Me, aminobutyric acid, Met, Val, pX-Phe (where X = F, Cl, Br, Oh , Or the D isomer of any one of Ala, Val, Gin, Asn, Leu, Ile, Met, pX (Wherein X = F, Cl, Br, O, CH, II or CI), Trp, Cys or -Nal; A 1-methyl-Nis, 3-methyl- that if Λ 1 - 0 1 Λ is present it can not be GGlu, and if A or A is present, it can not be pGlu; when A is eliminated and Ax is pClu, R 2 is II and R 0 has to be the portion of Glu that forms the inine nucleus in pGlu; and tnmhGr.1 - »where W can be; ) to the Cem which can be 1 or 0), or be eliminated, and 2, the identifier of the C-1, Ma, cyclochexyl-Ala, Vnl, Leu, Ile, 3er, Asp, Asn, Glu, Gin, p-X-Phe (wherein X-II,?, Cl, qr, -89/07 20 25 30 35 61.854 Ref: PTC / 3ioraeasure-3oabesin CIPS. 5-7; 9; 10 Cl), Trp, Cys, Vet, Pro, IlyPro or isopropyl, cyclohexylnoethyl, -Nal, -naphthylmethyl or phenylmethyl; and V is also wherein R 1 is any C 1 -C 4 alkyl, C ??? alkenyl, C 4 -C 4 alkynyl, naphthyl, or phenylalkyl, and each R 1 and R 2, independently 3-20 is selected from the group consisting of II, C1-4 alkyl, phenylalkyl lower acyloxy or wherein R3 is any of II, phenylalkyl or lower acyl); provided that when one of R or R g is NHR 3, the other is II; and provided that any asymmetric carbon atom may be R, s or a racemic mixture; and also provided that each Ib, and R4 * independently, is II, phenylalkyl alkyl, COE4 (wherein Εχ is C1-20 alkyl, C3-6 alkynyl, phenyl, naphthyl or phenylalkyl) R 2 and R 3 are attached to the terminal acid linker X of said peptide and provided that when any one of R 2 or R 2 is COE 2, the other has to be II, or a pharmaceutically acceptable salt thereof. 52. A process according to claim 4, wherein in the therapeutic peptide produced are: A = Gly, D-Phe, or deleted; A * = p-Glu, D-Phe, D-Ala, D-3-Nal, D-Cpa, or D-Asn; A 1 = Gin, 1-methyl, 1-methyl or 3-methyl-1 H; 4 Λ = Ala; A5 = Val; A-Sar, Gly, D-Phe or D-Ala; 7 A = Ilis; provided that where R for is the Leu or The identifying group; or where R0 is CIIof Σ1 is the group identifying [β-Leu or Leu; -40- 10 15 Mod. 71-10000 ex.-89/07 20 25 ΟΧ .OO-i Re: FTC / Uiomeasure-Ronbesin CIF3. 5-7; D; Or where Π 3 is C 1-10 -CH 2 Z is the Leu identifying group or its isopropyl, cyclohexylmethyl, B-naphthylmethyl or phenylalkyl; provided that where V is N, each R 2 and R 2 is II. or 63. A process according to claim 5, wherein in the therapeutic peptide produced V is ΝΙΙΓΙ and Π is 6 HH. 75. A process according to claim 5, wherein the therapeutic peptide produced is of the formula: pGlu-Cln-Trp-Ala-Val-Gly-His-Statin-amide. 83. A process according to claim 5, wherein the therapeutic peptide produced is of the formula: D-p-C1-Phe-Gln-Trp-Ala-Val-Gly-Ui s- [3-Leu-NHO. 9. A process as claimed in any one of the preceding claims comprising eight to ten amino acid residues, wherein said peptide is an analog of one of the following naturally occurring peptides terminating at the carboxy terminus of a residue Met: (a) litorin; (b) neurocmedin; (c) the zone of mammary gastrin-releasing peptide with ten amino acid residues at the r.arboxy terminal; (d) the amphibian bombesin zone with ten amino acids at the carboxy terminus, said peptide having the formula: ## STR1 ## wherein R 1, R 2, R 3, 30 Γ &gt; In which R 1, R 2, R 2, R 2, R 2, R 3 and R 4 are as defined above, (C 1 -C 8) alkyl, C 1 -C 6 alkyl, or C 3 -C 6 alkenyl, or to the group consisting of (a) or (b) of any of the pGIu, Nle or aridoc-aminobutyric groups; isomer V of any of Ala, Val, Gin, Asn, Leu, Ile, &quot;, 1.111 &gt; 35 10 Mod. 71 -10000 ex. - 89/07 20 Lof: FTC / Hiomeasure-Uom.bc-without CI 3 ♦ o-7; 2; (Ala, Vai, Gin, Asa, Cly, Ler., Ile, Mie, Azo-dobutyl, Met, pX-Phe (where X = 7, Cl, Hr, N * O, II, II or CIT "), Trp, Cys uu β-aai; (A-aminohutyric, Vet, Val pX-Phe (in which X = V, Cl, Cr, Ch, II or CII), Trp (Wherein X = Γ, Cl, Ur, h'Cy, R Ur, R Ur, R Ur, R Ur, R Ur, R Ur, 011, R1 or CH3), Trp, A * = 1-methylsilyl, 3-methylsulphonyl or Allyl if A is present, A can not be PgA, and if A or A 2 0 is present, A can not be pClu when it is deleted and A is pClu, P has to be II, and R has the portion of Clu which forms the imine ring in pGlu and also from YY may be: Ar, Gly, or Ha, :: et, pX-Cys, or [3-Nal V1 Z2 pX-Fhe (wherein X7, Cl, Dr, or C1-4 alkyl, C1-6 alkenyl, C1-6 alkenyl, C1-6 alkenyl, C1-4 alkenyl, C1-4 alkenyl, C1-6 alkenyl, 1, Cl, Cys, Xal, Ilis, 1 -, or 3-hydroxyethyl, N-CII-P , -CII-CV 46 where Π is C: 10 -HH, CIIO-3, CC-C10, or C1-4-C1-4, and each of Z1 and Z2 in each case is an identifier of any one of the amino acids Gly, Ala, Val, Leu , Ile, Ser, Asp, Asn, Glu, Cln, (¾-Hal, pX-Phe (wherein X-II, F, Cl, 3r, NOn CH or CII)), Trp, Cys, Vet, Pro, IlyPro , cyclohexyl-Ala; or cyclohexylmethyl; provided that where F 1 is CH 2 -Mi and Z 2 is the identifier group of any one of the amino acids Gly, Ala, Val, Leu, Ile, Ser, Asp, Asn, Clu, Gin, pX-Phe, wherein X = 1, F, Cl, 3r, NO ,, 011 or CIIO), Trp, Cys, Met, Pro, IlyPro, cyclohexylmethyl Z is the sole identifier of any of the amino acids Ser, Asp, Clu , Cys, Pro, HyPro or cyclohexylmethyl; and wherein R 1 is Cl 2 --HH and Z 2 is the identifier group of any one of the amino acids Gly, Ala, Val, Leu, Ile, Ser, Asp, Asn, Glu, Gin, pX-Fhe (wherein X = II, F, Cl, Πγ, ΝΌ, 01 !, 35 1 5 10 15 Mod. 71-10000 ex-89/07 20 25 30 oJL.SC54 King: PTC / Biomeasure-namingin CIFS 5-7; 9; 10 or CH 2), Trp, Cys, Met, Pio, or IlyPro, Z soya is the only group identifying any of the amino acids Ser, Asp, Glu, Cys, Pro, HyFro, or cyclohexylmethyl; and V is 0R "or Wherein each R 0, R 2, R 3 and R 4 independently are 1a, 1-lower alkyl, 1-phenyl-lower alkyl, or naphthyl-lower alkyl; and provided that any asymmetric carbon atom may be Π, S, or a racemic mixture; and also provided that each R 1 and R 2, independently, is II, phenyl C 1 -C 6 alkyl, COE (where E is C 1-6 alkyl, C 1-6 alkenyl, phenyl, naphthyl or phenylalkyl) or lower acyl, and H 2 and R 2 are attached to the N-terminal amino acid of said peptide, and also provided that when one of R 1 or R 2 is COE, the other is II, or a pharmaceutically acceptable salt thereof . 1. A process according to claim 9, wherein the therapeutic peptide produced comprises 0, A-Gly, D-Pbe or is deleted; Î ± = β-Glu, D-Phe, D-Ala, D-β-Nal, D-Cpa or D-Asn; 2 A = Gin, Ilis, 1-methyl-3 H or 3-methyl-3 '; 4A = Ala; A5 = Val; OAi =: 5a, Gly, D-Phe or D-Ala; λ '= His; wherein R is CH-Mi, each Z is cyclohexylmethyl or is the identifying group 4 1 of Leu or Phe; or Z0 is the identifying group of .alpha., Leu or Phe. 11. A process according to claim 9, wherein the therapeutic peptide produced A1 is D- (3-AT1), each of Z and Z2, independently, is either Leu or Phe12 ' (3MnI-Gln-Trp-Ala-Val-Cly-Ais-Leu [cII-Mg-Leu-NIi-13], wherein the therapeutic peptide produced has the formula: 3. A process according to claim 11, wherein the therapeutic peptide produced has the formula: -43 · 10 15 Mod. 71-10000ex.-89/07 20 25 30 · b64 Ref: PTC / 3iomeasure-3ombesin CIP 3, 5-7, 9, 10 D-piv-Mal-Gln-Trp-Ala-Val-Oly-IIis-Leu, 14. A process according to claim 9, wherein the peptide produced is CII, -NH and said carbon atom attached to Z has said R configuration. 155. A process according to claim 14, characterized in that the peptide produced is of the formula: wherein the peptide produced has the formula: A process according to claim 9, characterized in that in the therapeutic peptide prepared A ° or A * is an arsenic in the acid D and V is 0 R, 4 175. A process according to claim 16, The therapeutic peptide according to any one of claims 4 to 5, wherein the therapeutic peptide produced has the formula: D-Phe-Gln-Trp-Ala-Val-Cly-IIis-Leu-Met-methyl ether. amino acids, characterized in that said peptide is an analogue of the following naturally occurring peptides which terminate at the carboxy terminus with a residue Met: (a) (b) neuromedin, (c) the carboxy terminal amino acids of the carboxy terminal of the gastrin releasing peptide and (d) the ten amino acid region of the carboxy terminus of amphibian bombesin, therapeutic peptide of the formula: ## STR1 ## wherein R 1, R 2, R 3, R 4, In which R 1, R 2, R 2, R 2, R 2, R 2, R 2, R 2, R 2, R 2, R 2, R 2, , Cl, Pr, NCo, GII, II or Cil>, Trp, Cys, or -N-N1, which are deleted; Or the D-isomer of any one of Ala, Val, Cl-1, Asn, Leu, Ile Met, pX-? He (Gin which X = F, Cl, Pr, Mg, CH, H or C ln cil, C ln As, C ln cil cil, C ln cil cil, C ln cil cil, C ln cil cil, C ln cil, C ln cil cil, C ln cil cil, C ln cil, C ln cil, C ln cil, C ln cil, C ln cil, F, Cl, 3r, MC, OH, II or Cf),), Trp, Cys, FÒ-Nal, Mis, 1-methyl-IIis, or Reid: PTC / Dioneasure-Combssin CIPS. (A) Ala, Val, Gin, Asn, Gly, Leu, Ile, Nle, Î ± -aminobutyric acid, Vet, pX-Phe (wherein X = F, Cl, Br, N0o, O, II, or CILj), Trp, Cys, or -C (O) -Nal; Net, Val, pX-Phe, (wherein X-F, Cl, Sr, 011, II or CIF), Trp, Thr, or (3-Nal; 6 JA = Sar, Gly, or the D-isomer of any one of Ala, Val, Gin, Asn, Leu, He, Het, pX-Phe (wherein X = F, Cl , Dr, λΌ0, 0Π, I! or CIIO), Trp, Cys, or -Mal; A = 1-methyl-Kis, 3-methyl-Uis, or His; provided A is present, A ^ can not be pGlu; and if A₂ or s-2 is present, A can not be pGlu; and when A is eliminated, A is pGlu, R 2 has to be H and R 0 has to be the Glu portion which forms the amino imine in pGlu; and also since you ' can be; I 15 Mod. 71 -10000 ex. - 89/07 20 25 30 Wherein R 1, R 2, R 2, R 2, R 2, R 2, R 2, R 2, R 3, R 4, , Br, NO, OH or CH), Trp, Cys, Vet, Fro, or I and Pro; and each R11 and R3 is independently H, is lower alkyl, phenyl lower alkyl or naphthyl lower alkyl; and provided that any asymmetric carbon atom may be R, S or a racemic mixture; and also provided that each and Rn independently is II, C1-4 alkyl, C3-6 phenylalkyl, COE (wherein E is C1-6 alkyl, C1-6 alkenyl, C1-6 alkynyl, phenyl, naphthyl, or C7 -C10 phenylalkyl); or lower alkyl, and R 1 and S 2 are attached to the N-terminal amino acid of said peptide, and also provided that when either one of R 1 or R 2 is COE 2, the other has to be II, or a pharmaceutically acceptable salt thereof , 195. A process according to claim 18, wherein the therapeutic peptide produced is: CA = Gly, 9-Phe or deleted; A &quot; = p-Glu [?] - Phe, 9-Ala, D- [i-Nal, D-Cpa, or D-Asn; 2A = Gin, His, 1-methyl-Isis, or 3-methyl-Isis; Mod. 71-10000 ex. - 89/07 20 25 30 61,334 Ilef: PTU / Biomeasure-Bombesin CIP3. Î'-7; 9; 10 A-Ala; Λ5 = Go; A = Sar, Gly, D-Fhe, or D-Ala; - Hi a; provided that it is the identifying group of any of the amino acids Leu or D or L p -X-Fhe (wherein X = II, F, Cl, Cr, NOx or 011 or CH); and Hg, H10 and R3, independently, is II, lower alkyl, phenyl lower alkyl, or naphthyl lower alkyl. 20. A process according to claim 19, wherein in the produced peptide Z is Leu, R is II, and each R199C and R5 is lower alkyl. 21. A process according to claim 20, wherein the peptide produced has the formula: D-rhe-Gln-Trp-Gln-Gln-Gln-Leu-ethyl-d-a. 22. A process according to claim 20, wherein the therapeutic peptide produced is of the formula: D-Phe-Gln-Trp-Ala-Val-Gly-His-Leu-NII. The process of any preceding claim, comprising six to eight amino acid residues, inclusive, wherein the peptide produced is an analogue of one of the following naturally occurring peptides terminating in the carboxy-terminus or a residue I 'et: (&amp;)litorin; (b) neuromedin; (c) the ninth of the ten amino acids at the carboxy-terminus of mammary gastrin-releasing peptide; and (d) the amino acid zone of the carboxy-terminal amphibian of the hypobacterium, said peptide having the formula: ## STR2 ## which is 0, Gly, ..le, A?, A?, A?, As?, II? Ca,? or?,? 1 = the isomer D or L d? or the isomer, Ile, Ile, pX-Fhe (where X = F, Cl, 3r, Trp, or Bb-Mal, or is deleted, or any of pClu, or -amino acid, or the isomer D of any one of Ala, Val, Gin, Asn, Leu, Ile, -4G-35 10 15 Mod. 71 -10000 ex.- 89/07 20 25 30 35 31864 Hei: PTC / Diomeasure-Qombeain CIPS, 5-7: 9: 10 Met, p-X-Phe (where X = F, Cl, Br, N0o, 011, H or CH), Trp, Cys, <O or W-Nal, or is eliminated; (Wherein X = p, Cl, Br, NO0, CM, II or CI), Trp, Cys, β-Phexyl, Mal, Ilis, 1-methyl-His, or 3-methyl-Mis; A4 = Ala, Val, Gin, Asn, Gly, Leu, Ile, Mie, O (-aminobutyric acid, uet pX-Phe (where X = F, Cl, Br, M0o, H, or Cl) Trp, Cys, or β-Mal, - Gin, Asn, Gly, Ala, Leu, Ile, Nle, Î ± -aminobutyric acid, Met, Val, pX- rhe (wherein X = Î ±, Cl, Br, CH3, CH3, CH3 or CH3), Trp, The, or [5-Mal, A-3a, Cly, or the isomer of any one of Ala, Val, C1-6, Asn, Leu, Ile, Met, X-Phe (wherein X = F, Cl, Br, MC ,,, OH, II or CH, Trp, Cys, or (¾ -Mal: 70A-1-methyl-IIa, 3-methyl- or A 1, provided that if A is present, A 1 can not be pClu and if A or r 0 is present, it can not be pGlu when A is eliminated and is pGlu, has to be II and R 'has to be the moiety of the imine ring in pGlu, and also since W may be: wherein each R20 and R15, independently, and K, lower alkyl, phenyl-lower alkyl or naphthyl lower alkyl, provided that any asymmetric carbon atom may be R, S or a mixture of cycloalkenyl, and also provided that each and R ', independently is II, C1-4 alkyl, phenylalkyl, CCE, (wherein E, is C1-4 alkyl, alkenyl alkynyl, phenyl, naphthyl or phenylC_Q )alkyl), or lower acyl, and R e and R estejam are attached to the terminal amino acid M of said peptide, and also when one of COE ^ is the other has to be II, or a pharmaceutically acceptable salt thereof. 24. A process according to claim 23, wherein in the therapeutic peptide: A 1 = Gly, D-Phe or 4 is eliminated; A &quot; = p-Glu, D-Phe, D-Ala, D-Mal, D-Cpa, or D-Asn; -10-1 15 Mod. 71-lOOOOex. -89/07 20) 25 30 61,864 Pef; FTC / Hionensure-Bombesin CIPS. 5-7; 9; 10 2 A = Gin, His, 1-methyl-J is, or 3-methyl-Kis; A A ** = Ala; A5 = Val; A6 = Sar, Gly, D-Fue, or D-Ala; Λ '= His; wherein each R20 and R33 are II; and each and R 0, independently, are lower alkyl, or lower acyl. 25. A process according to claim 24, characterized in that in the therapeutic peptide produced &gt; on N are diols of II, A must be His, A must be Gly, A must be A process according to claim 24, characterized in that in the therapeutic peptide produced, R or are different from II, and A can not be removed. 27. A process according to claim 4, 9, 18 or 23, wherein the therapeutic peptide produced is. analogous to at least 25% homologous to said naturally occurring peptide, 285. A method according to claim 27, wherein said analog produced is at least 50% homologous to said naturally occurring peptide. 29. A process for the preparation of a therapeutic peptide bombesin according to the previous claims, characterized in that the peptide produced has the formula: p-Glu-Gln-Arg-Leu-Gly-Asn-Gln-Trp- Ala-Yal-Gly-Ile-Statin. 30. A composition according to any one of the preceding claims which comprises the preparation of an effective putative antagonistic peptide, wherein the peptide produced contains the amino acid having the formula: ^ -a1 !? / Rn-42- 35 10 15 Mod. 71-10000 ex. -89/07 20 Cl.864 P.ef; PTC / Biomeasure-Bonbesin CIPS. 5-7; 9; 10 .11 Λ2 13 14 A -A -A -A -N X R. in which A = pGlu, D or L, or is eliminated; 2 A = Gin, Asn, Gly, Ala, Leu, Ile, Nle, AccidocA -aninobutyric, Met, Val, Phe, pX-Phe (X = F, Cl, Br, OH or CH "), Trp, Naphthylalanine or is eliminated; 3A = Are ·, D-Arg, Lys, D-Lys or is deleted; 4A = Gin, Asn, Gly, Ala, Leu, Ile, Nle, Acidopaminobutyric, Vet, Val, Phe, pX-Phe, (X = F, Cl, Br, Oh or CU), Trp, β- or is eliminated; 5A = Gin, Asn, Gly, Ala, Leu, Ile, Nle, C acid &lt; (X = F, Cl, Br, OH or CH), Trp, (¾-naphthylalanine, D-Ala or is deleted; A6 = Oln, Asn, Gly, Ala, D-Ala, N-Ac-D-Ala, Leu, Ile, Nle, Amino-butyric acid, Et, Val, Phe, pX-rhe (X p -Glu, 1¾-naphthylanine or is eliminated; 7A-Cln, Asn, Gly, Ala, Leu, Ile, Nle, <X-aminolmutyric acid, Not, Val, Phe, D-Phe, pX-rhe (X = f, cl, Br, O , Cpd), Trp, Lys, Nis, or β-naphthylalanine, nobutyric, ethoxy, Phe, Phe, pX-rhe (X = F, Cl, Br, OH, CIII), Trp, SA-Trp or vet , Cln, Aán, Gly, Ala, Leu, Ile, Nle, Aminobutyric acid, Vet, Val, Tho, PX-Phe (X-F * t Br, OIT or Ci :,) Gn, Ala, Leu, Ile, Nle, Inovobutyric Acid, Net, Val, Fho, Γ-Χ-Phe , Phe, Phe, Phe, Phe, Phe, Phe, Phe, Phe, Phe, Phe, Phe, Phe, or p -H-Ph and (X = F, UJ., π D or 13-Cln, Asn, Gly, Ma, Leu, lio | (1-Amino-4-methoxyphenyl) -acetylamino] -acetylamino] -acetylamino] -acetic acid, , Trp, orfa-naphthylamine; A = Gin, Asn, Gly, Ala, Leu, Ile, Nle, Acetylamino-butyryl, Vot, Val, Phe, PX-Pho (X = F, Cl, Br, O11 or CII3), Trp or β -nafitlalanine; * ^ 19- 10 15 Mod. 71 -10000 βχ. - 89/07 20 25 30 € 1.864 He f; PTC / 3iome as Bonibe s in CIP3. 5-7; 9; R 1, R 2 and R 3 independently are each independently selected from the group consisting of C 1 -C 4 alkyl, C 1 -C 4 alkyl, phenyl, C 1 -C 4 alkyl, C 1 -C 20 alkynyl, naphthyl, or phenyl (C 1 -C 10) alkyl, or COOE (wherein C 6 alkyl or phenylalkyl and R 11 and R 11 are attached to the terminal amino acid M of said peptide, which may be A, A, A, A, A, or A *, is also provided that when any one of R or R is COE or C00S, the other has to be -O when one of K or R for is COE ou or COOE * the other must be II, and also provided that when A = = pGlu, R tenha has to be II and Π has to be the Glu moiety which forms the " 1] imine ring in pKlu; and for each of the residues A *, A, A, A, A and A are independently the carbon atom participating in the amide bond between that residue and the nitrogen atom of the group alpha amino acid of the contiguous amino acid residue may be a carbonyl carbon or may be reduced for a methylene carbon, provided that said at least one carbon atom can be reduced to a methylene carbon, said peptide further comprising A5-Cys6, A-Cys or an isomer D of any one of said amino acids; pGlu, Cys, 1-methyl-ilis, or 3-methyl-His; Cys; = Sar, or the D-isomer of any one of Ala, Val, Gin, Asn, Leu, He ITet, pX-Phe (wherein X = F, Cl, Lr, MO ,, H or CII), Trp, Cys or = 1-methyl-IIis, or 3-methyl-Tis; "14 and where A can be eliminated. The process according to claim 30, wherein the therapeutic peptide produced is of the formula: Dp-Cl-Phe-Gln-Trp-Ala-Val-Gly-Hi-Lou-Q: Phe-MH A = A9 to 11 .12 Lisbon, -2ΙΜΜ990 By ADMlMI STRATORS 0F THE TULAME EDUCATIONAL FUND. Inducing the Inducing Membrane, 3, M-1100 LMMM