WO1990006937A1

WO1990006937A1 - Derivatives of tetrapeptides as cck agonists

Info

Publication number: WO1990006937A1
Application number: PCT/US1989/005673
Authority: WO
Inventors: Kazumi Shiosaki; Alex M. Nadzan; Hana Kopecka; Youe-Kong Shue
Original assignee: Abbott Laboratories
Priority date: 1988-12-21
Filing date: 1989-12-18
Publication date: 1990-06-28
Also published as: JPH04502323A; CA2006189A1; EP0449884A4; GR890100827A; EP0449884A1

Abstract

Tetrapeptide analogs are disclosed which possess CCK agonist activity.

Description

DERIVATIVES OF TETRAPEPTIDES AS CCK AGONTSTS

This is a continuation-in-part of U.S. Patent

Application Serial No. 287,955, filed December 21, 1988.

Technical Field

The present invention relates to novel organic compounds and compositions which mimic the effects of cholecystokinin, caerulein and gastrin, processes for making such compounds, synthetic intermediates employed in these processes and a method for treating gastrointestinal disorders, central nervous system disorders, insulin related disorders, or potentiating pain, or regulating appetite with such compounds.

Background of the Inyention

Cholecystokinin (CCK) is a 39 amino acid polypeptide hormone. CCK and a 33 amino acid fragment of CCK (CCK₃₃) were first isolated from hog intestine (Mutt and Jorpes, Biochem. J . 125 628 (1981)). Recently the CCK₃₃ fragment has been found in the brain, where it appears to be the precursor of two smaller fragments, an octapeptide CCK₈ and a tetrapeptide CCK₄ (Dockray, Nature 264 402 (1979)). Existence of these fragments in the cortex of the brain suggests that CCK may be an important neuromodulator of memory, learning and control of the primary sensory and motor functions. CCK and its fragments are believed to play an important role in appetite regulation and satiety (Della-Fera, Science 206 471 (1979); Saito et al., Nature 289 599 (1981); and Smith, Eating and Its Disorders, eds., Raven Press, New York, 67 (1984)). Recently, patients with bulimia were shown to have lower than normal CCK levels in their plasma (Geracioti, et al., New England Journal of Medicine, 319 683 (1988) ). An additional role for CCK in the periphery is to regulate the release of insulin. CCK has been shown to increase the levels of insulin when administered to mammals (Rushakoff, et al., J. Clin. Endocrinol. Metab. 65 395 (1987)).

C-terminal fragments of CCK have recently been reported to function as CCK receptor antagonists (Jensen et al Biochem. Biophys. Acta, 757, 250 (1983); Spanarkel, J. Biol. Chem. 258. 6746 (1983)). Japanese patent

application 45/10506 to Miyao, et al., discloses a

tetrapeptide derivative of the carboxy terminal sequence of gastrin (L-Trp-L-Lys-L-Asp-L-Phe-NH₂) which has

antigastrin activity.

In contrast, the present invention relates to tetrapeptide analogs which function as agonists of CCK activity. CCK agonists are useful in the treatment and prevention of CCK-related disorders of the

gastrointestinal, appetite (obesity and bulimia, among others) and insulin regulatory systems of animals, especially man. CCK agonists are also useful as central nervous system suppressants which can exhibit anti-psychotic, neuroleptic, anxiolytic, and anti-convulsant effects, among other effects on central nervous system disorders.

Brief Description of the Drawings

Figure 1 is a plot comparing the mean level of liquid food intake (mis) for rats after chronic administration of vehicle, CCK-8 (10 nmol/kg), or the compound of Example 180 (1 nmol/kg or 10 nm/kg).

Figure 2 is a plot comparing the mean change in body weight (grams) for rats after chronic administration of vehicle, CCK-8 (10 nmol/kg), or the compound of Example 180 (1 nmol/kg or 10 nm/kg).

Summary of the Inyention

In accordance with the present invention there are cholecystokinin agonists of the formula:

Y - X - Z - Q

I

wherein Y is

or

wherein

(a) A is an N-protecting group and

B is -N(R₂₁)- wherein R₂₁ is hydrogen or loweralkyl; or wherein A is absent and B is halogen, cyano, or

-OR₂₂ wherein R₂₂ is hydrogen, alkanoyl,

alkoxycarbonyl or loweralkyl;

U is -CH-;

L is -CH₂-;

R₁ is bicyclic carbocyclic or bicyclic heterocyclic; and

R₂₀ is -C (O) - or -CH₂-;

or

(b) A and B are absent;

U and L taken together are an alkylene or alkenylene group;

R₁ is bicyclic carbocyclic or bicyclic heterocyclic; and

R₂₀ is -C(O)- or -CH₂-;

or

(c) A is absent;

B is absent or when U is N then B is hydrogen,

loweralkyl or -(CH₂)_xOR₂₃ wherein x is 2-6 and R₂₃ is hydrogen, loweralkyl, aryl or heterocyclic;

U is N, O or S;

L is -CH₂-; R₁ is bicyclic carbocyclic or bicyclic heterocyclic; and

R₂₀ is -C(=R₂₄)- wherein R₂₄ is O, S, NH or NR₂₅ wherein R₂₅ is loweralkyl;

or

(d) A is alkoxy, acyloxy, bicyclic carbocyclic or

bicyclic heterocyclic;

B is an alkylene group;

U is CH;

L is an alkylene group;

R₁ is alkoxy, acyloxy, bicyclic carbocyclic or bicyclic heterocyclic;

and R₂₀ is -C(O)- or -CH₂-;

or

(e) A and B are absent;

U is -CH₂-;

L is O, S or -N(R₂₆)- wherein R₂₆ is hydrogen or

loweralkyl;

R₁ is bicyclic carbocyclic or bicyclic heterocyclic; and

R₂₀ is -C(O)- or -CH₂-;

X is

wherein V is C₁ to C₆ alkylene or C₂ to C₆

alkenylene;

R₂₇ is hydrogen or loweralkyl;

T is -C(O)- or -CH₂-;

E is O, S, -N(R₃₀)- wherein R₃₀ is hydrogen or

loweralkyl, or E is -CH₂C(O)-, -C (O) CH₂-,

-CH₂CH₂-, -CH=CH-, -C(=R₂₁)R₂₃-, -R₂₂C(=R₂₁)- or -R₂₃C(=R₂₄)R₂₃- wherein R₂₁ is O or S, R₂₂ is N(R_2R), 0 or S, R₂₃ is independently selected at each occurrence from N(R₂₉), 0 and S, and R₂₄ is O, S or NH, wherein R₂₈ and R₂₉ are

independently selected from hydrogen and

loweralkyl; with the proviso that when R₂₂ is NH and V is alkenylene then the NH is not directly bonded to the double bond of the alkenylene group; and

R₂ is -GR₇ wherein G is absent, C₁ to C₄ alkylene, C₂ to C₄ alkenylene or a C₁ to C₄ alkylene or C₂ to C₄ alkenylene group which is substituted by a cyano group or an N-protected amino group and R₇ is loweralkyl, C₁ to C₁₂ alkenyl, adamantyl, aryl, arylalkyl, heterocyclic, cycloalkyl, substituted loweralkyl wherein the loweralkyl group is substituted with alkoxy, thioalkoxy, halo or N-protected amino or R₇ is substituted cycloalkyl wherein the cycloalkyl ring is substituted with one to four substituents independently selected from loweralkyl, halo and alkoxy; with the proviso that when E is -NHC(O)NH- then G is absent and with the proviso that when R₂₃ is NH and G is C₂ to C₄ alkenylene then the NH is not directly bonded to the double bond of the alkenylene group;

Z is

or

wherein R₃ is -CH₂CO₂R₃₂ wherein R₃₂ is hydrogen or loweralkyl, or R₃ is

R₃₁ is hydrogen or loweralkyl; R₂₅ is -C (O) - or

-CH₂-; and R₈₁ is -OR₈₂ wherein R₈₂ is hydrogen or loweralkyl or R₈₁ is

and

Q is

wherein R₄ is cyclohexyl, loweralkyl, aryl,

arylalkyl, heterocyclic or (heterocyclic) alkyl; R₅ is hydrogen or loweralkyl;

R₃₃ is -C(O)-, -C(S)- or -CH₂-; and

D is -NR₃₄R₈₀ wherein R₃₄ is hydrogen, hydroxy or

loweralkyl and R₈₀ is hydrogen or loweralkyl, or

D is -SR₃₅ wherein R₃₅ is hydrogen or

loweralkyl, or D is -OR₃₆- wherein R₃₆ is

hydrogen, loweralkyl or alkanoyl;

or a pharmaceutically acceptable salt thereof.

The term "loweralkyl" as used herein refers to

straight or branched chain alkyl radicals having from 1 to

7 carbon atoms including, but not limited to, methyl,

ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl,

pentyl, hexyl, heptyl and the like.

The term "cycloalkyl" as used herein refers to an aliphatic ring having 3 to 7 carbon atoms including, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like.

The term "alkenyl" as used herein refers to a C₁ to

C₁₂ straight or branched chain of carbon atoms which

contains a carbon-carbon double bond, such as allyl,

propenyl, butenyl, isoprenyl and the like. The term "alkylene" as used herein refers to a 1 to 6 carbon straight or branched chain di-radical including, but not limited to, -CH₂-, -CH(CH₃)-, -CH₂CH₂CH₂-,

-CH(CH₃)CH₂CH₂- and the like.

The term "alkenylene" as used herein refers to a 2 to 6 carbon straight or branched chain di-radical which contains a carbon-carbon double bond including, but not limited to, -CH=CH-, -CH=CHCH₂-, -CH=CHCH (CH₃) -,

-C(CH₃)=CHCH₂-, -CH₂CH=CHCH₂- and the like.

The term "halo" or "halogen" as used herein refers to chloro, bromo, iodo or fluoro.

The term "haloalkyl" as used herein refers to a loweralkyl radical in which one hydrogen atom has been replaced by a halogen including, but not limited to, chloromethyl, 2-fluoroethyl and the like.

The term "polyhaloalkyl" refers to a loweralkyl radical in which 2 or more hydrogen atoms have been replaced by halogens including, but not limited to

trifluoromethyl, dichloroethyl and the like.

The term "alkoxy" as used herein refers to -OR₈ wherein R₈ is loweralkyl.

The term "thioalkoxy" as used herein refers to -SR₉ wherein R₉ is loweralkyl.

The term "acyloxy" as used herein refers to -OC(O)R₁₀ wherein R₁₀ is loweralkyl.

The term "alkanoyl" asused herein refers to R₈₃C(O)- wherein R₈₃ is a loweralkyl group.

The term "alkoxycarbonyl" as used herein refers to R₈₄C(O)- wherein R₈₄ is an alkoxy group.

The term "aryloxy" as used herein refers to R₈₅O- wherein R₈₅ is an aryl group. The term "alkylamino" as used herein refers to -NHR₁₁ wherein R₁₁ is loweralkyl.

The term "dialkylamino" as used herein refers to - NR₁₂R₁₃ wherein R₁₂ and R₁₃ are independently selected from loweralkyl.

The term "bicyclic carbocyclic" as used herein refers to a group having two fused carbocyclic rings, each ring having 5, 6 or 7 carbon atoms, and each ring being fully saturated, partially saturated or aromatic. Bicyclic carbocyclic groups include, but are not limited to, naphthyl, tetrahydronaphthyl, decalin, indanyl, indenyl and the like. Bicyclic carbocyclic groups can be

unsubstituted or substituted with one, two or three substituents independently selected from loweralkyl, haloalkyl, polyhaloalkyl, alkoxy, thioalkoxy, aryloxy, benzyloxy, alkoxycarbonyl, alkanoyl, acyloxy, amino, alkylamino, dialkylamino, hydroxy, halo, mercapto, nitro, -OSO₃H, carboxaldehyde, carboxy and carboxamide.

The term "aryl" as used herein refers to a monocyclic or bicyclic carbocyclic ring system having one or more aromatic rings including, but not limited to, phenyl, naphthyl, tetrahydronaphthyl, indanyl, indenyl and the like; or "aryl" refers to a heterocyclic aromatic ring as defined below. Aryl groups can be unsubstituted or substituted with one, two or three substituents

independently selected from loweralkyl, haloalkyl, polyhaloalkyl, alkoxy, thioalkoxy, aryloxy, benzyloxy, alkoxycarbonyl, alkanoyl, acyloxy, amino, alkylamino, dialkylamino, hydroxy, halo, mercapto, nitro, -OSO₃H, carboxaldehyde, carboxy and carboxamide. The term "arylalkyl" as used herein refers to an aryl

group appended to a loweralkyl radical including, but not

limited to, benzyl, phenethyl, naphthylmethyl and the

like.

The term "heterocyclic group" or "heterocyclic" as used herein refers to any 3- or 4-membered ring containing a heteroatom

selected from oxygen, nitrogen and sulfur, or a 5- or 6-membered ring containing from one to three heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur; wherein the 5- membered ring has 0-2 double bonds and the 6-membered ring has 0-3 double bonds; wherein the nitrogen and sulfur heteroatoms may optionally be oxidized; wherein the nitrogen heteroatom may

optionally be quaternized; and including any bicyclic group in which any of the above heterocyclic rings is fused to a benzene ring or another 5- or 6-membered heterocyclic ring independently as defined above. Heterocyclics include, but are not limited to, quinolyl, isoquinolyl, indolyl, benzofuryl, benzothienyl, pyridyl, imidazolyl, furyl, thienyl, pyrazinyl, pyrrolyl, pyrimidyl and the like .

Heterocyclics can be unsubstituted or monosubstituted or disubstituted with substitutents independently selected from hydroxy, halo, oxo (=0), amino, alkylamino, dialkylamino, alkoxy, thioalkoxy, carboxy, alkoxycarbonyl, loweralkyl, cycloalkyl,

-OSO₃H, polyhaloalkyl and haloalkyl.

The term " (heterocyclic) alkyl" as used herein refers

to a heterocyclic group appended to a loweralkyl radical

including, but not limited to, quinolylmethyl,

imidazolylmethyl, benzimidazolylmethyl and the like.

The term "bicyclic heterocyclic" as used herein

refers to a group having two fused rings, one or both of

which are heterocyclic rings as defined herein. When both rings are not heterocyclic, the other ring is carbocyclic and is saturated, partially saturated or aromatic, preferably a fused benzene ring. Bicyclic heterocyclic groups can be unsubstituted or monosubstituted or

disubstituted with substitutents independently selected from hydroxy, halo, oxo (=0), amino, alkylamino,

dialkylamino, alkoxy, thioalkoxy, carboxy, alkoxycarbonyl, loweralkyl, cycloalkyl, -OSO₃H, polyhaloalkyl and

haloalkyl.

The term "N-protected" or "N-protecting group" as used herein refers to those groups intended to protect the N-terminus of an amino acid or peptide or to protect an amino group against undesirable reactions during a synthetic procedure or to prevent the attack of

exopeptidases on the compounds or to increase the

solubility of the compounds and includes, but is not limited to, sulfonyl, acetyl, pivaloyl, t-butyloxycarbonyl (Boc), benzyloxycarbonyl (Cbz), benzoyl or an L- or D- aminoacyl residue, which may itself be N-protected similarly.

All amino acid residues identified herein are in the natural L-configuration unless otherwise designated with "D-", (e.g., D-Trp). In keeping with standard peptide nomenclature, J. Biol. Chem.. 243:3557-59, (1969), abbreviations for amino acid residues are used herein. Abbreviations used herein are as shown in the following Table of Correspondence:

TABLE OF CORRESPONDENCE

SYMBOL REPRESENTS

Tyr L-tyrosine

Gly glycine

Phe L-phenylalanine

Met L-methionine

Ala L-alanine

Ser L-serine

lle L-isoleucine

Leu L-leucine

Thr L-threonine

Val L-valine

Pro L-proline

Lys L-lysine

Orn L-ornithine

His L-histidine

Gln L-glutamine

Glu L-glutamic acid

Trp L-tryptophan

Arg L-arginine

Asp L-aspartic acid

Asn L-asparagine

Cys L-cysteine

alpha-Nal alpha-naphthyl- alanine

beta-Nal beta-naphthylalanine

Ctp

The abbreviation ψ(CH₂NH) indicates that the amide (-C(O)NH-) bond of a peptide has been replaced by the reduced form -CH₂NH-. For example, Trpψ(CH₂NH) Lys represents a tryptophan residue bonded to a lysine residue wherein the amide bond is reduced as shown below.

It is noted that all amino acid residue sequences are represented herein by formulae whose left to right orientation is in the conventional direction of amino-terminus to carboxy-terminus.

Exemplary compounds of the present invention include:

t-butoxycarbonyl-Trp-Lys(epsilon-amino- (3-phenylpropionyl))-Asp-Phe-NH₂;

(t-butoxycarbonyl-beta-naphthyl-Ala)-Lys(epsilon-amino-(3-(4-hydroxyphenyl)propionyl))-Asp-Phe-NH₂; t-butoxycarbonyl-Trp-Lys(epsilon-N-(3(4-OSO₃H-phenyl)propionyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(3-carboxyquinolyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(4-hydroxyphenylacetyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(4-hydroxycinnamoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(4-OSO-H-cinnamoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(3-(3-hydroxyphenyl)propionyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(3-(3-OSO₃H-phenyl)propionyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(3-(4-chlorophenyl)propionyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(4-phenylbutyryl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(3-(4-methoxyphenyl)propionyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(3-(4-methylphenyl)propionyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(3-hydroxycinnamoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(3-OSO₃H-cinnamoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(4-methylcinnamoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(4-fluorocinnamoyl))-Asp-Phe-NH_?; t-butoxycarbonyl-Trp-Lys(epsilon-N-(4-trifluoromethylcinnamoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(3-(3-pyridyl)acrylyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(3-(4-fluorophenyl)propionyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(3-(4-trifluoromethylphenyl)propionyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(3-(3-indolyl)acrylyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(1-naphthoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(3-(2-thienyl)acrylyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(2-naphthoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(4-chlorocinnamoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(3-(3,4-dihydroxyphenyl)propionyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(6-acetoxy-2-naphthoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(alphacyano-3-hydroxycinnamoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N- (cinnamoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(1-adamantanoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(1-adamantaneacetyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(4- methoxycinnamoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(4- dimethylaminocinnamoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(4-bromocinnamoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(6-hydroxy-2-naphthoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(2,4-dichlorocinnamoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(4-nitrocinnamoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(3,4-dimethoxycinnamoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(4-(3-quinolyl)-3-butenoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(3,4-dihydroxycinnamoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(3,4-dichlorocinnamoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-D-Trp-Lys(epsilon-N-(3-(4-hydroxyphenyl)propionyl))-Asp]-Phe-NH₂;

t-butoxycarbonyl-D-Trp-Lys (epsilon-N- (4- hydroxycinnamoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-alpha-Nal-Lys(epsilon-N-(3- (4-hydroxyphenyl)propionyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-alpha-Nal-Lys(epsilon-N-(4-hydroxycinnamoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-beta-Nal-Lys(epsilon-N-(3-(4-hydroxyphenyl)propionyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-beta-Nal-Lys(epsilon-N-(4-hydroxycinnamoyl))-Asp-Phe-NH₂; t-butoxycarbonyl-Trp-Lys(epsilon-N-(3-(4-hydroxyphenyl)propionyl))-Asp-(NMe)Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(4-hydroxycinnamoyl))-Asp-(NMe)Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(4-chlorocinnamoyl))-Asp-(NMe)Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(6-hydroxy-2-naphthoyl))-Asp-(NMe)Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(6-acetoxy-2-naphthoyl))-Asp-(NMe)Phe-NH₂;

t-butoxycarbonyl-D-Trp-Lys(epsilon-N-(3-(4-hydroxyphenyl)propionyl))-Asp-(NMe)phe-NH₂;

t-butoxycarbonyl-D-Trp-Lys(epsilon-N-(4-hydroxycinnamoyl))-Asp-(NMe)Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(3-(4-hydroxyphenyl)propionyl))-Asp-alpha-Nal-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(2-hydroxycinnamoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(3-chlorocinnamoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-((6-OSO₃H-2-naphthoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(2,4-dimethoxycinnamoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(4-(2-naphthyl)-3-butenoyl))-Asp-Phe-NH₂;

Ctp-Lys(epsilon-N-(3-(4-hydroxyphenyl)-propionyl))-Asp-Phe-NH₂;

2-Naphthoxyacetyl-Lys(epsilon-N-(4-hydroxycinnamoyl))-Asp-Phe-NH₂; 3-(3-Indolyl)propionyl-Lys(epsilon-N-(3-(4- hydroxyphenyl)propionyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(3-(3- indolyl)propionyl)-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(Boc-Tyrosyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(Boc-O-sulfatyl-Tyrosyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(Boc-Trytophyl))-Asp-Phe-NH₂;

t-butoxycarbσnyl-Trp-alpha-aminopimelicacid(epsilon-Tyramide)-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(2-methylphenylaminocarbonyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(3-cyclohexylpropionyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(8-hydroxyquinolyl-2-carbonyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(5-methoxyindolyl-2-carbonyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-Boc-D-Tryptophyl)-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-Boc-D-Tyrosy1)-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(5-(benzyloxy)indole-2-carbonyl)Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(5-chloroindole-2-carbonyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(5-hydroxyindole-2-carbonyl))-Asp-Phe-NH₂; t-butoxycarbonyl-Trp-Lys(epsilon-N-(3-methylphenylaminocarbonyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(4-chlorophenylaminocarbonyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(2-chlorophenylaminocarbonyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(1-naphthylaminocarbonyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(phenylaminocarbonyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N- (cyclohexylaminocarbonyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(3-chlorophenylaminocarbonyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(3-methylphenylaminothiocarbonyl))-Asp-Phe-NH,;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(t-butylaminocarbonyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(2-methylphenylaminocarbonyl))-Asp-(NMe)Phe-NH₂;

t-butoxycarbonyl-Trpψ(CH,NH)Lys(epsilon-N-4-hydroxycinnamoyl)-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-4-hydroxycinnamoyl)ψ(CH,NH)Asp-Phe-NH₂.

BOC-Trp-Lys(ε-N-[2-nitrophenylaminocarbonyl])-Asp-Phe-NH_2;

BOC-Trp-Lys(ε-N-[2-triflurormethylphenylaminocarbonyl])-Asp-Phe- NH₂;

BOC-Trp-Lys(ε-N-[2-bromophenylaminocarbonyl])-Asp-Phe-NH_2;

BOC-Trp-Lys(e-N-[2-chlorophenylaminothiocarbonyl])-Asp-Phe-NH₂; BOC-Trp-Lys(ε-N-[2-methylphenylaminothiocarbonyl])-Asp- Phe-NH_2;

BOC-Trp-Lys(ε-N-[3-acetylphenylaminocarbonyl])-Asp-Phe-NH_2;

BOC-Trp-Lys(ε-N-[4-acetylphenylaminocarbonyl])-Asp-Phe-NH_2;

BOC-Trp-Lys(ε-N-[4-phenoxyphenylaminocarbonyl) ]-Asp-Phe-NH_2;

BOC-Trp-Lys(ε-N-[2-isopropylphenylaminocarbonyl])-Asp-Phe-NH₂;

BOC-Trp-Lys(ε-N-[S-2(1-naphthyl)ethylaminocarbonyl])- Asp-Phe-NH_2;

BOC-Trp-Lys(ε-N-[4-methylphenylaminocarbonyl])-Asp-Phe-NH_2;

BOC-Trp-Lys(ε-N-[2-methoxyphenylaminocarbonyl])-Asp-Phe-NH₂;

BOC-Trp-Lys(ε-N-[2-naphthyl])-Asp-Phe-NH₂;

BOC-Trp-Lys(ε-N-[2-(methoxycarbonyl)phenylaminocarbonyl])-Asp-Phe- NH_2;

BOC-Trp-Lys(ε-N-[3-(methoxycarbonyl)phenylaminocarbonyl])-Asp-Phe- NH_2;

BOC-Trp-Lys(ε-N-[2,6-dichlorophenylaminocarbonyl])- Asp-Phe-NH_2;

BOC-Trp-Lys(ε-N-[2,6-dimethylphenylaminocarbonyl])- Asp-Phe-NH_2;

BOC-Trp-Lys(ε-N-[allylaminocarbonyl])-Asp-Phe-NH₂;

BOC-Trp-Lys(ε-N-[4-nitrophenylaminocarbonyl])-Asp-Phe-NH_2;

BOC-Trp-Lys(ε-N-[benzylaminocarbonyl])-Asp-Phe-NH₂

BOC-Trp-Lys(e-N-[2-methylphenylaminocarbonyl])- Asp-HPhe-NH₂;

BOC-Trp-Lys(ε-N-[2-methylphenylaminocarbonyl])- Asp-Phenylalaninol; BOC-Trp-Lys(ε-N-[2-methylphenylaminocarbonyl])-Asp- NMe-PhenylaIaninol;

(Isobutoxycarbonyl)indolelactoyl-Lys[ε-N-(2-methylphenylaminocarbonyl])-Asp-Phe-NH_2;

Indolelactoyl-Lys(ε-N-[2-methylphenylaminocarbonyl])-Asp-Phe-NH₂;

BOC-Trp-Lys(ε-N-[4-hydroxycinnamoyl])-Asp-Trp-NH_2;

BOC-Trp-Orn(δ-N-[2-methylphenylaminocarbonyl])-Asp-Phe-NH_2; BOC-Trp-Orn(δ-N-[4-hydroxycinnamoyl])-Asp-Phe-NH_2;

BOC-Trp-HLys(ω-N-[2-methylphenylaminocarbonyl])-Asp- Phe-NH₂;

BOC-Trp-HLys(ω-N-Cbz)-Asp-Phe-NH₂;

BOC-Trp-HLys(ω-N-[4-hydroxyphenylcinnamoyl])-Asp-

Phe-NH₂;

BOC-Trp-Lys(ε-N-[3-pyridyl-3-acrylyl])-Asp- (NMe)Phe-NH_2;

BOC-Trp-(6-amino-1-[4-hydroxyphenethylamido]-hept-2-enoyl)-Asp-Phe-NH₂;

BOC-Trp-Lys(ε-N-[4-sulphatyl-cinnamoyl])-Asp-(NMe)Phe-NH_2;BOC-Trp-Lys(ε-N-[2-methylphenylaminocarbonyl])-Asp(β-Me)-Phe-NH₂;

Trp-Lys-(ε-N-[2-methylphenylaminocarbonyl])-Asp- (NMe)Phe-NH₂ Hydrochloride;

D-Trp-Lys-(ε-N-[2-methylphenylaminocarbonyl])-Asp- (NMe)Phe-NH₂ Hydrochloride;

BOC-Trp-Lys(ε-N-[2-methylphenylaminocarbonyl])-β-Asp- Phe-NH₂;

Ac-Trp-Lys(ε-N-[2-methylphenylaminocarbonyl])-Asp-Phe-NH₂; BOC-Trp-(NMe)Lys(ε-N-[2-methylphenylaminocarbonyl])-Asp-(NMe)Phe-NH₂; BOC-Trp- (NMe ) Lys (ε-N- [ 2-methylphenylaminocarbonyl ] ) - Asp-Phe-NH₂;

BOC-Trp-Lys(ε-N-[2-methylphenylaminocarbonyl])-Asp- Phe-NMe₂;

BOC-Trp-Lys(ε-N-[2-methylphenylaminocarbonyl])- (NMe)Asp-Phe-NH_2;

BOC-Trp-Lys(ε-N-[2-methylphenylaminocarbonyl])- (NMe)Asp-(NMe)Phe-NH_2;

BOC-Trp-Lys(ε-N-[2-methylphenylaminocarbonyl])-Asp-Phe-NHMe;

BOC-Trp-Lys(ε-N-[2-methylphenylaminocarbonyl])[ψCH₂NH]- Asp-Phe-NH_2;

2-Fluoro-3-(indol-3-yl)-propionyl-Lys(ε-N-[2-methylphenylaminocarbonyl])-Asρ-PheNH₂;

2-Cyano-3-(indol-3-yl)-propionyl-Lys(ε-N-[2-methylphenylaminocarbonyl])-Asp-PheNH₂; and

Boc-Trp-Lys(ε-N-[2-methylphenylaminocarbonyl])-Asp-Phe-OMe.

Preferred compounds of the invention include:

BOC-Trp-Lys(ε-N-[2-methylphenylaminocarbonyl])- (NMe) Asp-Phe-NH₂.

BOC-Trp-Lys(ε-N-[2-methylphenylaminocarbonyl])- (NMe)Asp-(NMe)Phe-NH₂;

BOC-Trp-Lys(ε-N-[2-methylphenylaminocarbonyl])- [ψCH₂NH] -Asp-Phe-NH_2;

Ac-Trp-Lys(ε-N-[2-methylphenylaminocarbonyl])-Asp-Phe-

NH₂;

Trp-Lys-(ε-N-[2-methylphenylaminocarbonyl])-Asp-

(NMe) Phe-NH₂ Hydrochloride;

Indolelactoyl-Lys(ε-N-[2-methylphenylaminocarbonyl])-

Asp-Phe-NH₂; BOC-Trp-Lys (ε-N- [2-methylphenylaminocarbonyl ] ) -Asp- Phenylalaninol;

(Isobutoxycarbonyl) indolelactoyl-Lys [ε-N- (2-methylphenylaminocarbonyl] ) -Asρ-Phe-NH₂;

BOC-Trp-Lys (ε-N- [2-bromophenylaminocarbonyl ] ) -Asp-Phe- NH_{2 ;}

t-butoxycarbonyl-beta-Nal-Lys(epsilon-N-(4-hydroxycinnamoyl))-Asp-Phe-NH,;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(4-hydroxycinnamoyl))-Asp-(NMe)Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(6-hydroxy-2-naphthoyl))-Asp-(NMe)Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(6-acetoxy-2-naphthoyl))-Asp-(NMe)Phe-NH₂;

t-butoxycarbonyl-D-Trp-Lys(epsilon-N-(4-hydroxycinnamoyl))-Asp-(NMe)Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-((6-OSO₃H-2-naphthoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(5-hydroxyindole-2-carbonyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(1-naphthylaminocarbonyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(2-methylphenylaminocarbonyl))-Asp-(NMe)Phe-NH₂; and t-butoxycarbonyl-Trp-Lys(epsilon-N-4- hydroxycinnamoyl)ψ(CH₂NH)Asp-Phe-NH₂.

The compounds of the present invention,

represented by formula I, can be prepared via a number of processes which have been developed for peptide synthesis. A detailed description of these methods is contained in "The Peptides, Vol. 1", Gross and

Meinenhofer, Eds., Academic Press, New York, 1979.

Coupling methods employed include the carbodiimide method (1,3-dicyclohexylcarbodiimide [DCC] , 1-(3-dimethylaminoproρyl-3-ethylcarbodiimide hydrochloride [EDCI]) with the option of racemization preventing additives (1-hydroxybenzotriazole [HOBT] ) , the mixed anhydride method, the azide method, the acid chloride method, the symmetrical anhydride method, the use of bis (2-oxo-3-oxazolidinyl)phosphinic chloride (BOP-Cl), and the active ester method (N-hydroxysuccinimide esters, 4-nitrophenol esters, 2,4,5-trichlorophenol esters, and the like).

The compounds of the invention are prepared by stepwise coupling of the amino acids or by coupling together fragments of dipeptide length or greater.

Thus, the free carboxylic acid moiety from one amino acid or peptide fragment is activated and allowed to condense with the free nitrogen group of the second amino acid or peptide fragment. The coupling reactions are conducted in solvents such as methylene chloride (CH₂Cl₂), tetrahydrofuran (THF), dimethylformamide (DMF) or other such solvents under an inert atmosphere such as nitrogen (N₂) or argon (Ar). During the coupling process, the non-participating carboxylic acids or amines on the reacting set of amino acids or peptide fragments are protected by a

protecting group which can be selectively removed at a later time if desired. A detailed description of these groups and their selection and chemistry is contained in "The Peptides, Vol. 3", Gross and Meinenhofer, Eds., Academic Press, New York, 1981. Thus, useful

protective groups for the amino group are

benzyloxycarbonyl (Cbz), t-butyloxycarbonyl (BOC) , 2,2,2-trichloroethoxycarbonyl (Troc), t-amyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 2- (trichlorosilyl) ethoxycarbonyl, 9-fluorenylmethoxycarbonyl (FMOC) , phthaloyl, acetyl (Ac), formyl, trifluoroacetyl, and the like.

Examples of useful protective groups for the carboxylic acid include esters such as methyl, ethyl, benzyl, t-butyl, 2,2,2-trichloroethyl, allyl, 4-nitrobenzyl, and the like. Removal of these protecting groups can be accomplished selectively by employing various acid or base catalyzed hydrolytic,

hydrogenolytic, thermal or dissolving metal conditions.

For the production of a compound of the invention where any one or several of the constituent amino acids bear an N-alkyl group, specifically methyl, the

corresponding N-alkyl amino acid can be prepared via the method described by Benoiton (Can. J. Chem., 55, 906 (1977)) or Shuman ("Peptides: Proceedings of the 7th American Peptide Symposium", D. Rich, E. Gross, Eds., Pierce Chemical Co., Rockford, IL 1981, p 617) wherein the Boc or Cbz protected amino acid is treated with a base in the presence of a chelating agent such as a crown ether and then quenched with methyl iodide. An alternative method described by Freidinger ( J. Org. Chem., 48, 77 (1983)) in which triethylsilane reduction of the oxazolidinone of an amino acid directly produces the N-methyl derivative can also be utilized.

The reduced carbonyl amide bond surrogates can be prepared in a manner similar to that described by

Martinez (J. Med. Chem. 30 1366 (1987)). The N-alpha-Boc protected amino acid (with appropriate protection of side chain functional groups) is converted to the 3,5-dimethylpyrazolide, which is then reduced with lithium aluminum hydride. The resulting aldehyde is then allowed to condense with an amino acid or peptide bearing a free amino terminus. Reduction of the Schiff base which is formed as a result of the condensation is accomplished using sodium cyanoborohydride to yield the desired compound having a reduced amide bond.

Functionalization of the epsilon-amino group of the Lys or homologous (e.g., Orn) residue is achieved via activation of the acid fragment as the active ester (N-hydroxysuccinimide, 2,4,5-trichlorophenol, etc.) or, if no other free carboxylic acid function is present on the peptide, coupling using any of the methods

mentioned above is applicable. In addition, the functionalization of the epsilon-amino group can be accomplished by reaction with various alkyl and aryl isocyanates, as well as alkyl and aryl isothiocyanates.

The sulfuric acid esterification of the phenolic residues can be conducted using a variety of known reagents such as the pyridine-sulfuric anhydride or the pyridine-sulfur trioxide complex. Use of pyridinium acetyl sulfate as described by Penke and Rivier

("Proceedings of the 8th American Peptide Symposium", V. Hruby, D. Rich, Eds., Pierce Chemical Company,

Rockford, IL.; 1983; p. 119) can also be applied to prepare the sulfuric acid ester derivative of the tetrapeptides.

The following examples will serve to further illustrate preparation of the novel compounds of the invention.

Example 1

BOC-Asp (beta-Bn)-Phe-NH₂

To a solution of phenylalanineamide hydrochloride (19.4 g, 0.06 mol) in dimethylformamide (100 mL) cooled to 0ºC were added N-methylmorpholine (7.2 mL, 0.065 mol), a solution of BOC-Asp b-benzyl ester (12.0 g, 0.06 mol) in methylene chloride (80 mL), 1-hydroxybenzotriazole (12.2 g, 0.09 mol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI) (12.4 g, 0.065 mol). The reaction was stirred overnight with warming to ambient temperature. The solvent was removed in vacuo and the resulting residue was dissolved in ethyl acetate and washed with 1 M H₃PO₄ (3X) , saturated sodium bicarbonate (NaHCO₃) solution (3X) and brine. After drying (MgSO₄), the solvent was evaporated and the residue was dissolved in hot ethyl acetate and the product precipitated with dropwise addition of hexane. The product was collected and dried to yield 25 g of a white solid. MS(CI/NH₃) m/e 470 (M+H) ⁺, 487. ¹H NMR(CDCl₃, 300MHz) δ 1.39 (s,9H), 2.78 (dd, J=18Hz,1H), 2.92-3.05 (m,2H), 3.21 (dd,1H), 4.38-4.45 (m, 1H) , 4.65 (q, J=6Hz, 1H), 5.36 (br s,1H), 5.49 (br d, J=7.5Hz, 1H), 6.09 (br s,1H), 6.82 (br d,J=7hz,1H), 7.21-7.40 (m, 10H) .

Example 2

ASP (beta-Bn)-Phe-NH-Hydrochloride A solution of example 1 (16.2 g, 34 mmol) in 50 mL of 1.5 M HCl (g) in acetic acid was stirred at ambient temperature for 1.5 h. The reaction was quenched with the addition of ether which precipitated the product. The solid was collected, washed with fresh ether and dried to yield 12.9 g of white powder. MS(CI/NH₃) m/e 370 (M+H)⁺. ¹H NMR(DMSO-d6, 300MHz) δ 2.74-2.91 (m, 2H), 2.98-3.09 (m,2H), 4.08-4.12 (m, 1H) , 4.42-4.51 (m, 1H), 5.17 (br s,2H), 7.14-7.59 (m, 10H), 8.26 (br s,1H), 8.71 (br d, J=7Hz,1H).

Example 3

BOC-Lys (epsilon-N-Cbz)-ASP(beta-Bn)-Phe-NH₂

To a solution of the hydrochloride of example 2 (12.9 g, 32 mmol) in dimethylformamide (20 mL) and methylene chloride (20 mL) cooled to -10 C were added N-methylmorpholine (3.9 mL, 35 mmol), BOC-Lys (epsilon-N-Cbz) (12.1g, 32 mmol), 1-hydroxybenzotriazole (6.5 g, 48 mmol), and EDCI (6.7g, 35 mmol). The reaction was stirred overnight with warming to ambient temperature. The solvents were removed in vacuo and the residue was dissolved in ethyl acetate and washed successively with solutions of 1M H₃PO₄ (3X), saturated NaHCO₃ (3X) and brine. The solvent was removed in vacuo and the solid residue was dissolved in acetone with warming. The product was precipitated with the dropwise addition of water, collected and dried to yield 22.3 g of a white powder. MS(CI/NH₃) m/e 732 (M+H)⁺, 749, 632. ¹H

NMR(DMSO-d6,300 MHz) δ 1.13-1.54 (m, 6H), 1.37 (br

s,9H), 2.51-3.05 (m, 6H) , 3.86 (br s,1H), 4.38 (br

s,1H), 4.61 (br s,1H), 5.00 (s,2H), 5.07 (s,2H), 6.87 (br d, J=7Hz,1H), 7.12-7.38 (m, 16H) , 7.85 (br d,1H), 8.15 (br d,1H). Anal calcd for C₃₉H₄₉N₄O₉: c 64.00,H 6.75,N 9.57; found: C 63.92, H 6.82, N 9.54.

Example 4

Lys(epsilon-N-Cbz)-ASP(beta-Bn)-Phe-NH₂ Hydrochloride A solution of example 3 (19 g, 26 mmol) in 80 mL of 1.5 M HCl (g) in acetic acid was stirred at ambient temperature for 1.5 h. The product was precipitated with the addition of ether (1 L), collected and dried to yield 17.2 g of a white powder. MS(CI/NH₃) m/e 632 (M+H)⁺. ¹H NMR(DMSO-d6, 300MHz) δ 1.21-1.43 (m, 4H), 1.59-1.70 (m, 2H), 2.59-2.69 (m,4H), 2.78-3.06 (m, 5H), 3.69 (br s,1H), 4.36-4.45 (m,1H), 4.62-4.72 (m, 1H), 5.00 (br s,2H), 5.10 (br s,2H), 7.11-7.40 (m,16H), 8.17 (br d, J=7Hz, 1H), 8.76 (br d, J=7Hz,1H) .

Example 5

BOC-Trp-Lys(epsilon-N-Cbz)-ASP(beta-Bn)-Phe-NH

To a solution of the hydrochloride salt of example 4 (9.8 g, 15.5 mmol) in dimethylformamide (100 mL) cooled to 0°C were added N-methylmorpholine (1.9 mL, 17 mmol) and BOC-Trp N-hydroxysuccinimide ester (6.15 g, 15.5 mmol). The reaction was stirred overnight with warming to ambient temperature. The solvent was removed in vacuo and the residue partitioned between a solution of citric acid and ethyl acetate. The organic phase was further washed with solutions of NaHCO₃ (3X) and water (3X).

After drying with MgSO₄, the solvent was removed in vacuo and the solid residue dissolved in ethyl acetate/acetone and precipitated with the addition of water. The product was collected and dried to yield 11.9 g of a white solid. MS(FAB+) m/e 918 (M+H)⁺, 940 (M+Na)⁺. ¹H NMR (DMSO-dδ, 300MHz) δ 1.01-1.66 (m, 6H), 1.29 (br s, 9H), 2.52-3.31 (m,8H), 4.18-4.31(m,2H), 4.36-4.45 (m, 1H), 4.57-4.68 (m,1H), 4.99 (br s,2H), 5.06 (br s,2H), 6.82-7.39

(m, 20H), 7.58 (br d, J=7Hz, 1H), 7.82 (br d,1H), 7.92 (br S,1H), 8.29 (br d, J=7Hz, 1H), 10.78 (br s,1H). Anal calcd for C₅₀H₅₉N₇O₁₀ ·25H₂O: C 65.09, H 6.57, N 10.62; found: C 64.80, H 6.49, N 10.65. Example 6

BOC-Trp-Lys-Asp-Phe-NH₂

A mixture of example 5 (5.0 g, 5.45 mmol) and 10% Pd/C (1.0 g) in acetic acid (100 mL) was hydrogenated under one atmosphere of hydrogen at ambient temperature overnight. The catalyst was filtered and the solvent was removed in vacuo. The residue was triturated with ether to yield 3.95 g of a light pink powder. MS (FAB+) m/e 694 (M+H)⁺. ¹Η NMR(DMSO-d6, 300MHz) δ 1.11-1.62 (m, 6H) , 1.32 (br s,1H), 2.25-3.18 (m, 8H), 4.10-4.43 (m, 4H), 6.78-7.34 (m,l2H), 7.51-7.62 (m,2H), 8.05 (br d, J=7Hz, 1H), 8.17 (br d,J=7Hz,1H), 10.91 (br s,1H). Anal calcd for C₃₅H₄₇N₇O₈ 1.75CH₃CO₂H: C 57.88, H 6.81, N 12.27; found: C 57.84, H 6.92, N 12.64.

Example 7

3-Phenylpropionic acid N-hydroxysuccinimide ester

A solution of 3-phenylpropionic acid (300 mg, 2 mmol), N-hydroxysuccinimide (276 mg, 2.40 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI) (421 mg, 2.2 mmol) in methylene chloride (30 mL) was stirred at ambient temperature for 18 h. The solvent was removed in vacuo and the residue chromatographed (ethyl acetate/hexane) to yield 410 mg of a white solid. MS(CI/NH₃) m/e 248 (M+H)⁺, 265. ¹H NMR (CDCl₃, 300MHz) δ 2.84 (br s,4H), 2.90-2.98 (m, 2H), 3.03-3.10 (m,2H), 7.20-7.35 (m,5H).

Example 8

BOC-Trp-Lys (epsilon-N-(3-phenylpropionyl))-Asp-Phe-NH₂ To a solution of example 6 (75 mg, 0.11 mmol) in dimethylformamide (7 mL) cooled to 0 C were added N-methylmorpholine (18 mg, 0.18 mmol) and the active ester of example 7 (32 mg, 0.13 mmol). The mixture was stirred overnight with warming to ambient temperature. The DMF was removed in vacuo and the residue was chromatographed on silica gel using ethyl acetate/pyridine/acetic

acid/water (42/3.3/1/1.8). The solvents were removed in vacuo and the residue was dissolved in aqueous acetone and lyophilized to yield 48 mg of a white, flocculent solid. MS(FAB+) m/e 826 (M+H)⁺, 726. ¹H NMR(DMSO-d6,300MHz) δ 1.12-1.58 (m, 6H), 1.32 (br s,9H), 2.32-2.56 (m,4H), 2.78-3.20 (m, 8H), 4.18-4.28 (br s,2H), 4,32-4.38 (m, 1H), 4.43-4.50 (m, 1H), 6.72 (br s,1H), 6.92-7.35

(m, 12H), 7.48 (br S,1H), 7.58 (br d, 1H), 7.78 (br s,1H), 7.92 (br d, 1H), 8.09 (br s, 1H), 8.20 (br d, 1H), 8.29 (br s,1H), 10.86 (br s,1H). Anal calcd for C₄₄H₅₅N₇O₉

2CH₃CO₂H: C 60.94,H 6.71, N 10.36; found: C 60.72, H 6.45, N 10.72.

Example 9

BOC-Trp-Lys (epsilon-N-(3-(4-hydroxyphenyl)- propionyl))-Asp-Phe-NH₂

The tetrapeptide of example 6 (670 mg),

diisopropylethylamine (130 mg) and 3-(4-hydroxyphenyl)propionic acid N-hydroxysuccinimide ester (271 mg) in DMF (20mL) were reacted under similar

conditions to those described in example 8. The product was isolated and purified in a similar manner to yield 515 mg of a white flocculent solid. MS (FAB+) m/e 842 (M+H)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 1.12-1.58 (m, 6H) , 1.31 (br s,9H), 2.28 (br t,J=9Hz,2H), 2.40-2.52

(m,2H), 2.69 (t,J=9H,2H), 2.81-3.20 (m, 8H) , 4.23 (br s,2H), A . 36 (m,1H), 4.49 (m, 1H) , 6.63 (d, J=9Hz), 1H), 6.78 (br d, J=8Hz,1H), 6.91-7.38 (m, 10H) , 7.58 (br d, J=7Hz, 1H), 7.73 (br t,1H), 7.89-7.99 (m, 2H) , 8.23 (br d, J=7Hz.1H), 10.81 (br s,1H). Anal calcd for C₄₄H₅₅N₇O₁₀ H₂O: C 61.45, H 6.68, N 11.40; found: C 61.12, H 6.48, N 11.14.

Example 10

BOC-Trp-Lys(epsilon-N-(3-(4-OSO₃H-phenyl)propionyl))- Asp-Phe-NH₂

To a solution of example 9 (103 mg, 0.12 mmol) in dimethylformamide (4 mL) and pyridine (4 mL) was added pyridinium acetyl sulfate (270 mg, 1.23 mmol). The reacion was stirred at ambient temperature for 18 h then poured into water (50 mL) and made basic with 1 M NaOH solution to pH 7.0-7.5. The mixture was

concentrated in vacuo and the residue suspended in methanol and filtered. The filtrate was evaporated in vacuo and the residue chromatographed via preparative reverse phase HPLC using acetonitrile and 0.05 M

ammonium acetate buffer at pH 4.5 as eluants. The product was lyophilized to yield 52 mg of a white flocculent powder. MS(FAB+) m/e 922 (M+H)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 1.08-1.60 (m, 6H), 1.31 (br s,9H), 2.33 (t, J=9Hz,2H), 2.41-3.18 (m, 10H), 4.23 (br s,2H), 4.38 (m,1H), 4.50 (m, 1H), 6.80 (br s,1H), 6.91-7.33 (m,12H), 7.57 (br d, 1H), 7.78 (br d, 1H), 7.89 (br t,2H), 8.25 (m,1H), 10.66 (br s, 1H). Anal calcd for C₄₄H₅₅N₇O₁₃S NH₄OH: C 55.22, H 6.32, N 11.71; found: C 55.35, H 6.22, N 11.52.

Example 11

3-Ouinoline carboxylic acid N-hydroxysuccinimide ester

A solution of 3-quinolinic acid (300 mg), N-hydroxysuccinimide (245 mg) and EDCI (349 mg) in methylene chloride (20 mL) was stirred at ambient temperature for 18h. The product was isolated as described for example 7 to yield 380 mg of a white solid. MS(CI/NH₃) m/e 271 (M+H)⁺. ¹H

NMR(CDCl₃, 300MHz) δ 2.97 (br s,4H), 7.18-7.22 (m, 1H), 7.89-8.00 (m,2H), 8.22 (d, J=9Hz, 1H), 9.01 (m, 1H), 9.49 (d, J=2Hz,1H).

Example 12

BOC-Trp-Lys (epsilon-N-(3-carboxyquinolyl))- Asp-Phe-NH₂

The tetrapeptide of example 6 (60 mg), active ester of example 11 (28 mg) and N-methylmorpholine (9 mg) were reacted under similar conditions to those described in example 8. The product was isolated in a similar manner to yield 47 mg of a white solid. MS (FAB+) m/e 849 (M+H)⁺. ¹H NMR(DMSO-d6, 300MHz) δ 1.11-1.67 (m,6H), 1.39 (br s,9H), 2.45-2.68 (m, 4H), 2.81-3.18 (m,4H), 4.22-4.42 (m, 3H), 4.48-4.55 (m, 1H), 6.77 (br s,1H), 6.91-7.41 (m, 10H), 7.55-7.68 (m,2H), 7.82 (t, J=7Hz,1H), 7.91-8.09 (m, 4H), 8.28 (br s,1H), 8.82 (br s,1H), 9.29 (br s,1H), 10.80 (br s,1H). Anal calcd for C₄₅H₅₂N₈O₉: C 63.66, H 6.17, N 13.20; found: C 63.59, H 6.26, N 13.12. Example 13

4-Hydroxyphenylacetic acid N-hydroxysuccinimide ester

A solution of 4-hydroxyphenylacetic acid (300 mg), N-hydroxysuccinimide (272 mg) and EDCI (416 mg) in

methylene chloride (20 mL) was stirred at ambient

temperature for 18 h. The product was isolated as

described in example 7 to yield 360 mg of a white

solid. MS(CI/NH₃) m/e 250 (M+H)⁺. ¹H NMR

(CDCl₃,300MHz) δ 2.83 (br s,4H), 3.83 (s,2H), 6.82

(d, J=9Hz,2H), 7.14 (d, 2H).

Example 14

BOC-Trp-Lys (epsilon-N-(4-hydroxyphenylacetyl))- Asp-Phe-NH₂

The tetrapeptide of example 6 (175 mg), active ester of example 13 (75 mg) and N-methylmorpholine (28 mg) were reacted under similar conditions to those described in example 8. The product was isolated in a similar manner to yield 149 mg of a white solid. MS (FAB+) m/e 828

(M+H)⁺, 850 (M+Na)⁺. ¹H NMR(DMSO-d6, 300MHz) δ 1.12-1.62

(m,6H), 1.40 (br s,9H), 2.42-2.72 (m, 4H) , 2.81-3.16

(m,4H), 3.25 (s,2H), 4.23 (br s,2H), 4.32-4.41 (m, 1H),

4.48-4.55 (m,1H), 6.66 (d, J=8Hz, 2H), 6.80 (br d, J=9Hz, 1H),

6.91-7.34 (m, 11H), 7.58 (br d,J=9Hz, 1H), 7.82-7.92 (m,3H),

8.26 (br d,J=9Hz,1H), 9.18 (br s,1H), 10.68 (br s,1H).

Anal calcd for C₄₃H₅₃N₇O₁₀ 1.5H₂O: C 60.41, H 6.60, N

11.47; found: C 60.38, H 6.30, N 11.41.

Example 15

4-Hydroxycinnamic acid N-hydroxysuccinimide ester

A solution of 4-hydroxycinnamic acid (300 mg), N-hydroxysuccinimide (252 mg) and EDCI (385 mg) in methylene chloride (20 mL) was stirred at ambient temperature for 18 h. The product was isolated as described in example 7 to yield 280 mg of a white solid. MS(CI/NH₃) m/e 262 (M+H)⁺. 1H NMR(DMSO-d6, 300MHz) δ 2.85 (br s,4H), 6.68

(d, J=15Hz,1H), 6.83 (d, J=8Hz, 2H), 7.68 (d,2H), 7.85

(d, 2H).

Example 16

BOC-Trp-Lys (epsilon-N-(4-hydroxycinnamoyl))-Asp-Phe-NH₂

The tetrapeptide of example 6 (120 mg), active ester of example 15 (60 mg) and N-methylmorpholine (20 mg) were reacted under similar conditions to those described in example 8. The product was isolated in a similar manner to yield 96 mg of a white flocculent powder. MS (FAB+) m/e 840 (M+H)⁺. ¹H NMR(DMSO-d6, 300MHz) δ 1.12-1.68

(m, 6H), 1.32 (br s, 9H) , 2.38-2.68 (m, 4H) , 2.82-3.19 (m, 4H), 4.19-4.31 (m,2H), 4.32-4.41 (m, 1H), 4.45-4.53 (m, 1H), 6.41 (d, J=15Hz,1H), 6.76 (d, J=8Hz, 2H), 6.92-7.45 (m, 13H), 7.58 (d, J=7Hz,1H), 7.92-8.05 (m, 3H) , 8.25 (br d, J=7Hz, 1H), 10.82 (br s,1H). Anal calcd for C₄₄H₅₃N₇O₁₀: C 62.92, H 6.36, N 11.67; found: C 63.24, H 6.43, N 11.64.

Example 17

BOC-Trp-Lys (epsilon-N-(4-OSO₃H-cinnnamoyl))-Asp-Phe-NH₂

The tetrapeptide of example 16 (50 mg) was treated in a manner similar to example 10 employing pyridinium acetyl sulfate (131 mg) in a solution of DMF and pyridine. The product was isolated and purified under identical

conditions to yield 34 mg of a white solid. MS (FAB-) m/e

918 (M-H)-. ¹H NMR(DMSO-d6, 300MHz) δ 1.12-1.18 (m, 6H),

1.32 (br s,9H), 2.45-2.75 (m, 4H) , 2.82-3.24 (m,4H), 4.21- 4.32 (m, 2H), 4.38-4.44 (m, 1H), 4.49-4.58 (m, 1H), 6.51

(d, J=15Hz,1H), 6.78 (br s,1H), 6.92-7.46 (m,16H), 7.58 (br d,J=7Hz,1H), 7.82 (br d,J=7Hz, 1H), 7.92 (br d, J=7Hz, 1H),

8.01 (br s,1H), 8.27 (br s,1H), 10.72 (br s, 1H). Anal calcd for C₄₄H₅₅N₇O₁₃S NH₄OH: C 53.81, H 6.26, N 11.41; found: C 53.81, H 5.79, N 11.24.

Example 18

3-(3-Hydroxyphenyl)propionic acid

N-hydroxysuccinimide

A solution of 3-(3-hydroxyphenyl)propionic acid (300 mg), N-hydroxysuccinimide (270 mg) and EDCI

(415 mg) in methylene chloride (20 mL) was stirred at ambient temperature for 18 h. The product was isolated as described in example 7 to yield 276 mg of a white solid. MS(CI/NH₃) m/e 264 (M+H)⁺. ¹H

NMR(CDCl₃, 300MHz) δ 2.81 (br s,4H), 2.81-2.98 (m, 4H), 6.58-6.72 (m,3H), 7.06 (t, J=7Hz, 1H), 9.28 (br s,1H).

Example 19

BOC-Trp-Lys (epsilon-N-(3-(3-hydroxyphenyl)- propionyl))-Asp-Phe-NH₂

The tetrapeptide of example 6 (120 mg), active ester of example 18 (60 mg) and N-methylmorpholine (20 mg) were reacted under similar conditions to those described in example 8. The product was isolated in a similar manner to yield 126 mg of a white solid. MS(FAB+) m/e 864 (M+Na)⁺. ¹H NMR (DMSO-d6, 300MHz) δ

1.12-1.62 (m,6H), 1.32 (br s,9H), 2.32 (t, J=7Hz, 2H),

2.42-2.67 (m,2H), 2.71 (t,2H), 2.80-3.23 (m, 6H) , 4.24 (br s,2H), 4.38 (br s,1H), 4.52 (m, 1H) , 6.52-6.61

(m,2H), 6.82 (br s,1H), 6.92-7.35 (m, 12H), 7.58 (br d,J=7Hz,1H), 7.75 (br s,1H), 7.84 (br d, J=7Hz, 1H), 7.92 (br d,J=7Hz,1H), 8.22-8.30 (m, 1H) , 9.19 (br s, 1H),

10.75 (br s,1H). Anal calc for C₄₄H₅₅N₇O₁₀ 1.5H₂O: C

60.81, H 6.72, N 11.28; found: C 60.70, H 6.49, N

11.33.

Example 20

BOC-Trp-Lys (epsilon-N-(3-(3-OSO₃H-phenyl)- propionyl))-Asp-Phe-NH₂

The tetrapeptide of example 19 (50 mg) was treated in a similar manner as described for example 10 employing pyridinium acetyl sulfate (130 mg) in a solution of DMF and pyridine. The product was isolated and purified in an identical manner to yield 21 mg of a white solid. MS (FAB-) m/e 920 (M-H)-. ¹H NMR (DMSO-d 6,300MHz) δ 1.12-1.62 (m, 6H),

1.32 (br s,9H), 2.34 (t, J=7Hz, 2H), 2.45-3.16 (m, 10H), 4.23

(br s,2H), 4.39 (br s,1H), 4.51 (m, 1H) , 6.82-7.35 (m,16H),

7.57 (br d, J=7Hz,1H), 7.82 (br d, J=7Hz, 2H), 7.90 (br d,J=7Hz,1H), 8.23-8.29 (m, 1H), 10.74 (br s,1H). Anal calcd for C₄₄H₅₅N₇O₁₅S NH₄OH: C 54.70, H 6.36, N 11.60; found:

C 54.61, H 6.21, N 11.62.

Example 21

3-(4-Chlorophenyl)propionic acid

N-hydroxysuccinimide ester

A solution of 3-(4-chlorophenyl)propionic acid

(300 mg), N-hydroxysuccinimide (225 mg) and EDCI (344 mg) in methylene chloride (20 mL) was stirred at

ambient temperature for 18 h. The product was isolated as described in example 7 to yield 330 mg of a white solid. MS(CI/NH₃) m/e 281 M⁺, 299 (M+NH₄)⁺. ¹H NMR (CDCl₃, 300MHz) δ 2.81 (br s,4H), 2.89-3.05 (m, 4H), 7.32 (br s,5H).

Example 22

BOC-Trp-Lys (epsilon-N-(3-(4-chlorophenyl)propionyl))- Asp-Phe-NH₂

The tetrapeptide of example 6 (55 mg), active ester of example 21 (29 mg) and N-methylmorpholine (9 mg) were reacted under similar conditions to those described in example 8. The product was

chromatographed and isolated in a similar

manner to yield 43 mg of a white solid. MS(FAB+) m/e 860 (M+H)⁺, 882 (M+Na)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 1.10-1.64 (m,6H), 1.31 (br s,9H), 2.35 (t, J=7Hz, 2H), 2.42-2.71 (m,4H), 2.76-3.15 (m, 6H), 4.24 (br s,2H), 4.34-4.42 (m,1H), 4.48-4.56 (m, 1H), 6.80 (d, J=9Hz, 1H) , 6.92-7.35 (m,15H), 7.58 (d, J=9Hz, 1H), 7.76 (br t,1H), 7.35-7.44 (m,2H), 8.24 (d, 1H), 10.78 (br s,1H). Anal calcd for

C₄₄H₅₄ClN₇O₉ 1.5H₂O: C 59-55, H 6.47, N 11.05; found: C 59.60, H 6.22, N 10.99. Example 23

4-Phenylbutyric acid N-hydroxysuccinimide ester

A solution of 4-phenylbutyric acid (300 mg), N-hydroxysuccinimide (252 mg) and EDCI (385 mg) in methylene chloride (20 mL) was stirred at ambient temperature for 18 h. The product was isolated as described in example 7 to yield 340 mg of a clear oil. MS(CI/NH₃) m/e 262 (M+H)⁺, 279 (M+NH₄)⁺. ¹H NMR (CDCl₃, 300MHz) δ 2.02-2.14 (m,2H), 2.61 (t, J=8Hz,2H), 2.74 (t, J=8Hz, 2H), 2.85 (br s,4H), 7.17-7.34 (m,5H).

Example 24

BOC-Trp-Lys (epsilon-N-(4-phenylbutyryl))-Asp-Phe-NH₂ The tetrapeptide of example 6 (60 mg), active ester of example 23 (30 mg) and N-methylmorpholine (10 mg) were reacted under similar conditions to those described in example 8. The product was isolated in a similar manner to yield 55 mg of a white powder. MS (FAB+) m/e 840

(M+H)⁺, 862 (M+Na)⁺. ^"4*1 NMR (DMSO-d6, 300MHz) 5 1.10-1.63 (m,6H), 1.31 (br s, 9H), 1.72-1.85 (m,2H), 2.03-2.10

(m,2H), 2.38-3.17 (m, 10H), 4.19-4.31 (m, 2H), 4.34-4.42 (m,1H), 4.48-4.56 (m, 1H), 6.80 (br d, 1H), 6.92-7.34

(m, 13H), 7.58 (d, J=9Hz, 1H), 7.72 (br t,1H), 7.83 (br d,1H), 7.90 (br d, 1H), 8.23-8.30 (m,2H), 10.75 (br

s,1H). Anal calcd for C₄₅H₅₇N₇O₉ 0.5H₂O: C 63.66, H 6.89, N 11.55; found: C 63.59, H 6.79, N 11.53.

Example 25

3-(4-Methoxyphenyl)propionic acid

N-hydroxysuccinimide ester

A solution of 3-(4-methoxyphenyl) propionic acid (512 mg), N-hydroxysuccinimide (392 mg) and EDCI (599 mg) in methylene chloride (20 mL) was stirred at ambient temperature for 18 h. The product was isolated as described in example 7 to yield 650 mg of a white solid. MS(CI/NH₃) m/e 277 M⁺, 295 (M+NH₄)⁺. ¹H

NMR (CDCl₃, 300MHz) δ 2.82-2.93 (m,6H), 4.98-3.04 (m,2H), 3.29 (s,3H), 6.85 (dt, J=9Hz,2H), 7.15 (dt,2H).

Example 26

BOC-Trp-Lys (epsilon-N-(3-(4-methoxyphenyl)- propionyl))-Asp-Phe-NH₂

The tetrapeptide of example 6 (50 mg), active ester of example 25 (25 mg) and N-methylmorpholine (8 mg) were reacted under similar conditions to those described in example 8. The product was isolated in a similar manner to yield 31 mg of a white flocculent solid. MS(FAB+) m/e 878 (M+Na)⁺. ¹H NMR(DMSO-d6,300MHz) δ 1.12-1.61 (m, 6H), 1.32 (br s,9H), 2.25- 3.25 (m,12H), 3.70 (s,3H), 4.21-4.49 (m, 4H), 6.74-7.35 (m, 13H), 7.58 (br d, 1H), 7.29 (br s,2H), 7.95 (br d,1H), 8.13-8.30 (m, 2H), 11.00 (br s,1H). Anal calcd for C₄₅H₅₇N₇O₁₀ 0.5H₂O: C 62.48, H 6.76, N 11.34;

found: C 62.42, H 6.73, N 11.26.

Example 27

3-(4-Methylphenyl)propionic acid

N-hydroxysuccinimide ester

A solution of 3-(4-methylphenyl)propionic acid (300 mg), N-hydroxysuccinimide (253 mg) and EDCI (386 mg) in methylene chloride (25 mL) was stirred at ambient temperature for 18 h. The product was isolated in an identical manner to example 7 to yield 382 mg of a white solid. MS(CI/NH₃) m/e 262 (M+H)⁺. ¹H NMR (CDCl₃, 300MHz) δ 2.32 (s,3H), 2.84 (br s,4H), 2.86-2.93 (m,2H), 2.99-3.06 (m, 2H), 7.12 (s,5H).

Example 28

BOC-Trp-Lys (epsilon-N-(3-(4-methylphenyl)propionyl))- Asp-Phe-NH₂

The tetrapeptide of example 6 (50 mg), active ester of example 27 (23 mg) and N-methylmorpholine (8.3 mg) were reacted under similar conditions to those described in example 8. The product was purified and isolated in an identical manner yielding 26 mg of a white solid. MS(FAB+) m/e 840 (M+H)⁺. ¹H NMR(DMSO-d6,300MHz) δ 1.15-1.65 (m, 6H), 1.32 (br s,9H), 2.24 (s,3H), 2.18-3.18 (m, 12H), 4.15-4.45 (m, 4H), 6.61 (br d,1H), 6.90-7.32 (m, 12H), 7.57 (d, J=7Hz, 1H), 7.79-7.89 (m, 2H), 7.96 (br d,1H), 8.15 (br d, 1H), 11.08 (br s,1H). Anal calcd for C₄₅H₅₇N₇O₉: C 64.39, H 6.84, N 11.67; found: C 63,71, H 6.87, N 11.45.

Example 29

3-Hydroxycinnamic acid N-hydroxysuccinimide ester

The active ester was prepared in a similar manner to that described for example 7 using 3-hydroxycinnamic acid (300 mg), N-hydroxysuccinimide (252 mg) and EDCI (385 mg). The product was isolated as described to yield 375 mg of a white solid. MS(CI/NH₃) m/e 279

(M+NH₄)⁺. ¹H NMR(DMSO-d6, 300MHz) δ 2.85 (br s,4H), 6.76-6.96 (m,2H), 7.17 (br S,1H), 7.25 (br d, 2H), 7.87 (d, J=16Hz,1H).

Example 30

BOC-Trp-Lys (epsilon-N-(3-hydroxycinnamoyl))- Asp-Phe-NH₂

The tetrapeptide of example 6 (97 mg), active ester of example 29 (44 mg) and N-methylmorpholine (16 mg) were reacted under similar conditions to those described in example 8. The product was isolated in a similar manner to yield 85 mg of a white solid.

MS(FAB+) m/e 841 (M+H)⁺. ¹H NMR(DMSO-d6, 300MHz) δ

1.13-1.68 (m, 6H), 1.32 (br s,9H), 2.23-3.19 (m, 8H), 4.21-4.48 (m, 4H), 6.61 (d, J=16Hz,1H), 6.72-7.33

(m,13H), 7.57 (br d, J=8Hz, 1H), 7.82 (br s,1H), 7.99 (br d,1H), 8.19 (br d,2H), 8.50 (br s,1H), 11.01 (br s, 1H). Anal calcd for C₄₄H₅₃N₇O₁₀ : C 62.92, H 6.36, N 11.67; found: C 62.59, H 6.40, N 11.54.

Example 31

BOC-Trp-Lys (epsilon-N-(3-OSO₃H-cinnamoyl))-Asp-Phe-NH₂ The tetrapeptide of example 30 (30 mg) was treated under similar conditions as described for example 10 employing pyridinium acetyl sulfate (78 mg) in pyridine and DMF. The product was isolated in an identical manner to yield 13 mg of a white, flocculent solid. MS(FAB+) m/e

996 (M+KI)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 1.11-1.66 (m, 6H) ,

1.31 (br s,9H), 2.25-3.18 (m, 8H) , 4.18-4.31 (m,2H), 4.33- 4.42 (m,1H), 4.48-4.56 (m, 1H) , 6.59 (d, 16HZ, 1H), 6.31-7.44

(m,14H), 7.58 (d, J=8Hz, 1H) , 7.85 (br d, 1H), 7.92 (br d, 1H), 8.15 (br s,2H), 10.78 (br s, 1H). Anal calcd for C₄₄H₅₅N₇O₁₃S 2H₂O 0.5NH₄OH: C 54.28, H 6.16, N 10.79; found: C 54.02, H 5.81, N 10.80.

Example 32

4-Methylcinnamic acid N-hydroxysuccinimide ester

A solution of 4-methylcinnamic acid (500 mg), N- hydroxysuccinimide (425 mg) and EDCI (650 mg) in methylene chloride (20 mL) was stirred at ambient temperature for 18 h. The active ester was isolated as described in example 7 to yield 680 mg of a white solid. MS(CI/NH₃) m/e 260 (M+H)⁺, 277 (M+NH₄)⁺. ¹H NMR (CDCl₃, 300MHz) δ 2.38 (s,3H), 2.86 (br s,4H), 6.89 (d, J-16.5Hz, 1H) , 7.28 (d, J=9Hz, 2H), 7.72 (d,2H), 7.92 (d, 1H).

Example 33

BOC-Trp-Lys (epsilon-N-(4-methylcinnamoyl))-Asp-Phe-NH₂

The tetrapeptide of example 6 (75 mg), active ester of example 32 (34 mg) and N-methylmorpholine (12 mg) were allowed to react under similar conditions to those

described in example 8. The final product was isolated in a similar manner to yield 48 mg of a white flocculent solid. MS(FAB+) m/e 860 (M+Na)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 1.12-1.68 (m,6H), 1.31 (br s, 9H), 2.32 (s,3H), 2.38-2.69 (m,2H), 2.80-3.20 (m, 6H), 4.26 (br s,2H), 4.32-4.42

(m,1H), 4.45-4.55 (m, 1H), 6.57 (d, J=16.5Hz, 1H), 6.79 (br d, J=9Hz,1H), 6.92-7.43 (m,16H), 7.50 (br d, 1H), 7.92-8.00 (m, 2H), 8.05 (br t,1H), 8.26 (br d, 1H) , 10.81 (br s,1H). Anal calcd for C₄₅H₅₇N₇O₁₀ H₂O: C 63.14, H 6.71, N 11.45; found: C 63.06, H 6.43, N 11.34.

Example 34

4-Fluorocinnamic acid N-hydroxysuccinimide ester

A solution of 4-fluorocinnamic acid (500 mg), N-hydroxysuccinimide (415 mg) and EDCI (634 mg) in methylene chloride (20 mL) was stirred at ambient temperature for 18 h. The product was chromatographed and isolated as described in example 7 to yield 680 mg of a white solid. MS(CI/NH₃) m/e 281 (M+NH₄)⁺. ¹H NMR(CDCl₃,300MHz) δ 2.87 (br s,4H), 6.95 (d,J=15Hz, 1H), 7.30 (t, J=10Hz, 2H), 7.91-8.01 (m,3H).

Example 35

BOC-Trp-Lys (epsilon-N-(4-fluorocinnamoyl))-Asp-Phe-NH₂ The tetrapeptide of example 6 (75 mg) , active ester of example 34 (34 mg) and N-methylmorpholine (12 mg) were allowed to react under similar conditions as those described in example 8. The peptide was isolated in an identical manner to yield 50 mg of a white solid.

MS(FAB+) m/e 842 (M+H)⁺. ¹H NMR(DMSO-d6,300MHz) δ 1.08-1.67 (m,6H), 1.29 (br s,9H), 2.38-2.66 (m,2H), 2.79-3.18 (m, 6H), 4.17-4.40 (m, 3H) , 4.49 (q, J=7Hz, 1H), 6.58

(d, J=15Hz,1H), 6.80-7.45 (m,16H), 7.54-7.62 (m,2H), 7.90- 8.00 (m,2H), 8.11 (br t,1H), 8.26 (br d, 1H), 10.84 (br s , 1H) . Anal calcd for C₄₄H₅₂FN₇O₉ 1 . 5H₂O : C 60 . 82 , H 6 . 38 , N 11 . 28 ; found : C 60 . 56, H 6 . 09 , N 11 . 18 .

Example 36

4-Trifluoromethylcinnamic acid

N-hydroxysuccinimide ester

A solution of 4-trifluoromethylcinnamic acid (500 mg), N-hydroxysuccinimide (319 mg) and EDCI (488 mg) in methylene chloride (20 mL) was stirred at ambient

temperature for 18 h. The product was isolated as previously described in example 7 to yield 580 mg of a white solid. MS(CI/NH₃) m/e 331 (M+NH₄)⁺. ¹H

NMR (CDCl₃, 300MHz) δ 2.87 (br s, 4H), 7.15 (d, J=16Hz, 1H), 7.82 (d, J=7Hz,2H), 8.01-8.11 (m, 3H).

Example 37

BOC-Trp-Lys (epsilon-N-(4-trifluoromethylcinnamoyl))- Asp-Phe-NH₂

The tetrapeptide of example 6 (75 mg), active ester of example 36 (41 mg) and N-methylmorpholine (12 mg) were allowed to react under similar conditions to those described in example 8. The peptide was isolated in an identical manner to yield 64 mg of a white solid. MS (FAB+) m/e 892 (M+H)⁺. ¹H NMR(DMSO-d6, 300MHz) δ 1.05-1.65 (m,6h), 1.29 (br s, 9H), 2.36-2.65 (m, 2H), 2.78-3.75

(m, 6h), 4.16-4.41 (m, 3H), 4.48 (q, J=8Hz, 1H), 6.77

(d, J=16Hz,1H), 6.83 (d, J=9Hz, 1H), 6.92-7.34 (m, 15H), 7.48 (d, 1H), 7.59 (br d,1H), 7.92-8.03 (m,2H), 8.24 (br t,2H), 10.84 (br s,1H). Anal calcd for C₄₅H₅₂F₃N₇O₉ 1.5H20: C 58.81, H 6.03, N 10.67; found: C 58.60, H 5.86, N 10.65. Example 38

3-(3-Pyridyl)acrylic acid N-hydroxysuccinimide ester A solution of 3-(3-pyridyl)acrylic acid (500 mg), N-hydroxysuccinimide (463 mg), and EDCI (707 mg) in

methylene chloride (20 mL) and dimethylformamide (10 mL) was stirred at ambient temperature for 18 h. The product was isolated as described in example 7 to yield 740 mg of a white solid. MS (CI/NH₃) m/e 247 (M+H)⁺. 1H

NMR (CDCl₃, 300MHz) δ 2.87 (br s,4H), 7.15 (d, J=16Hz, 1H), 7.48-7.53 (m, 1H), 8.02 (d, 1H), 8.31 (dt, J=9Hz, 1H), 8.66 (dd, J=6Hz,1H), 8.99 (d, J=2Hz, 1H).

Example 39

BOC-Trp-Lys (eosilon-N-(3-(3-Pyridyl)acrylyl))-Asp-Phe-NH₂ The tetrapeptide of example 6 (75 mg), active ester of example 38 (32 mg) and N-methylmorpholine (12 mg) were allowed to react under similar conditions to those described in example 8. The peptide was isolated in the usual manner to yield 53 mg of a white solid. MS (FAB+) m/e 825 (M+H)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 1.09-1.67

(m,6H), 1.29 (br s,9H), 2.40-2.69 (m,2H), 2.79-3.20 (m, 6H), 4.16-4.41 (m, 3H) , 4.50 (q, J=7Hz, 1H), 6.62

(d, J=17Hz,1H), 6.83 (br d, J=6Hz, 1H), 6.91-7.48 (m, 15H), 7.58 (d, J=8Hz,1H), 7.86-7.95 (m, 2H), 8.17 (br t, 1H), 8.27 (br d,1H), 8.52 (dd, J=5Hz, 1H), 8.72 (br d, 1H), 10.80 (br s,1H). Anal calcd for C₄₃H₅₂N₈O₉ 1.5H₂O: C 60.62, H 6.51, N 13.15; found: C 60.70, H 6.23, N 13.05.

Example 40

3-(4-Fluorophenyl)propionic acid

N-hydroxysuccinimide ester

A solution of 3-(4-fluorophenyl)propionic acid (500 mg), N-hydroxysuccinimide (410 mg) and EDCI (630 mg) in methylene chloride (20 mL) was stirred at ambient

temperature for 18 h. The product was isolated as described in example 7 to yield 650 mg of a white solid. MS(CI/NH₃) m/e 266 (M+H)⁺. ¹H NMR (CDCl₃, 300MHz) δ 2.84 (br s,4H), 2.86-2.95 (m, 2H), 3.00-3.08 (m, 2H), 7.00

(tt, J=9Hz,2H), 7.15-7.23 (m,2H).

Example 41

BQC-Trp-Lys(epsilon-N-(3-(4-fluorophenyl)propionyl))- Asp-Phe-NH₂

The tetrapeptide of example 6 (70 mg), active ester of example 40 (32 mg) and N-methylmorpholine (11 mg) were allowed to react under similar conditions to those described in example 8. The peptide was isolated in a similar manner to yield 40 mg of a white, flocculent solid. MS(FAB+) m/e 844 (M+H)⁺. ¹H NMR(DMSO-d6, 300MHz) δ 1.08-1.62 (m,6H), 1.30 (br s, 9H), 2.30-2.66 (m, 4H), 2.74-3.16 (m,6H), 4.15-4.41 (m,3H), 4.49 (br q, 1H), 6.82 (br s, J=6Hz,1H), 6.92-7.45 (m,14H), 7.59 (br d, 1H), 7.79 (br t,1H), 7.90-8.04 (m,2H), 8.24 (br d, J=8Hz, 1H), 10.84 (br s,1H). Anal calcd for C₄₄H₅₄FN₇O₉ H₂O: C 61.31, H 6.55, N 11.37; found: C 60.96, H 6.25, N 11.25.

Example 42

3-(4-Trifluoromethylphenyl)propionic acid

N-hydroxysuccinmide ester

A solution of 3-(4-trifluoromethylphenyl) propionic acid (500 mg), N-hydroxysuccinimide (320 mg) and EDCI (480 mg) in methylene chloride (20 mL) was stirred at ambient temperature for 18 h. The product was isolated as described for example 7 to yield 570 mg of a white solid. MS(CI/NH₃) m/e 316 (M+H)⁺. ¹H NMR(CDCl₃, 300MHz) δ 2.85

(br s,4H), 2.95 (t, J=8Hz, 2H), 3.12 (t,2H), 7.36

(d, J=8Hz,2H), 7.58 (d,1H).

Example 43

BOC-Trp-Lys (epsilon-N-(3-(4-trifluoromethylphenyl)- propionyl))-Asp-Phe-NH₂

The tetrapeptide of example 6 (70 mg), active ester of example 42 (38 mg) and N-methylmorpholine (11 mg) were reacted under similar conditions to those described in example 8. The product was isolated in an identical manner to yield 44 mg of a white solid. MS (FAB+) m/e 916 (M+Na)⁺. ¹H NMR (DMSO-d6,300MHz) δ 1.08-1.62 (m, 6H), 1.29 (br s,9H), 2.35-2.68 (m, 4H) , 2.77-3.16 (m, 8H) , 4.16-4.30 (m, 2H), 4.30-4.41 (m, 1H), 4.49 (q, J=7Hz,1H), 6.82 (br d, J=7Hz,1H), 6.92-7.43 (m, 13H), 7.56-7.62 (m, 2H), 7.82 (br t,1H), 7.90-8.02 (m,2H), 8.24 (br d, 1H), 10.84 (br s,1H). Anal calcd for C₄₅H₅₄F₃N₇O₉ 0.5H₂O: C 58.17, H 5.85, N 10.44; found: C 58.10, H 5.61, N 10.34.

Example 44

3-(3-Indolyl)acrylic acid N-hydroxvsuccinimide ester

A solution of 3-(3-indolyl)acrylic acid (500 mg), N-hydroxysuccinimide (370 mg) and EDCI (560 mg) in methylene chloride (20 mL) and dimethylformamide (5 mL) was stirred at ambient temperature for 18 h. The active ester was isolated as described for example 7 to yield

450 mg of a light brown solid. MS(CI/NH₃) m/e 285 (M+H)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 2.89 (br s,4H), 6.57 (d, J=16Hz, 1H), 7.52 (m,1H), 7.95-8.20 (m, 4H), 12.09 (br s,1H).

Example 45

BOC-Trp-Lys (epsilon-N-(3-(3-indolyl)acrylyl))-Asp-Phe-NH₂ The tetrapeptide of example 6 (70 mg), active ester of example 44 (34 mg) and N-methylmorpholine (11 mg) were reacted under similar conditions to those described for example 8. The final product was isolated in an identical manner to produce 40 mg of a white solid. MS (FAB+) m/e 863 (M+H)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 1.08-1.68 (m, 6H), 1.29 (br s,9H), 2.38-2.68 (m,2H), 2.78-3.21 (m, 6H), 4.18-4.41 (m,3H), 4.50 (q, J=7Hz,1H), 6.62 (d, J=16Hz, 1H), 6.82 (br d, J=6Hz, 1H), 6.92-7.48 (m,10H), 7.56-7.64 (m, 2H) , 7.71 (br s,1H), 7.85-8.00 (m, 2H), 8.26 (br d, J=7Hz,1H), 10.83 (br s, 1H). Anal calcd for C₄₆H₅₄F₈N₉ 1.5H2O: C 62.08, H 6.46, N 12.59;

found: C 62.29, H 6.22, N 12.59.

Example 46

1-Naphthoic acid N-hydroxysuuccinimide ester A solution of 1-naphthoic acid (500 mg), N-hydroxysuccinimide (400 mg) and EDCI (610 mg) in methylene chloride (20 mL) was stirred at ambient temperature for 18 h. The active ester was isolated as described in example 7 to yield 660 mg of a white solid. MS(CI/NH₃) m/e 270

(M+H)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 2.95 (br s,4H), 7.18-7.32 (m, 3H), 8.15 (d, J=8Hz, 1H), 8.40 (t, J=9Hz, 2H), 8.65 (d, 1H).

Example 47

BOC-Trp-Lys (epsilon-N-(1-naphthoyl))-Asp-Phe-NH₂

The tetrapeptide of example 6 (70 mg), active ester of example 46 (33 mg) and N-methylmorpholine (11 mg) were allowed to react under similar conditions to those described in example 8. The peptide was isolated in an identical manner to yield 42 mg of a white, flocculent solid.

MS(FAB+) m/e 848 (M+H)⁺. ¹H NMR(DMSO-d6, 300MHz) δ 1.08-1.71 (m, 6H), 1.30 (br s,9H), 2.38-2.68 (m, 2H), 2.78-3.18 (m, 6H), 4.17-4.40 (m,3H), 4.46-4.55 (m, 1H) , 6.81 (d, J=7Hz, 1H), 6.92-7.62 (m, 10H), 7.92-8.02 (m, 2H), 8.16-8.29 (m,2H), 8.51 (br t,1H), 10.83 (br s,1H). Anal calcd for C₄₆H₅₃N₇O₉ H₂O 0.5CH₃CO₂H: C 63.00, H 6.41, N 10.94; found: C 62.93, H 6.15, N 11.16.

Example 43

3- (2-Thienyl)acrylic acid N-hydroxysuccinimide ester A solution of 3-(2-thienyl)acrylic acid (500 mg), N-hydroxysuccinimide (450 mg) and EDCI (680 mg) in methylene chloride (20 mL) was stirred at ambient temperature for 18 h. The active ester was isolated as described in example 7 to yield 630 mg of a white solid. MS(CI/NH₃) m/e 252

(M+H)⁺. ¹H NMR (CDCl₃, 300MHz) δ 2.87 (br s,4H), 6.37

(d,J=16Hz,1H), 7.11 (m,1H), 7.38 (d, J=3Hz, 1H), 7.50

(d, J=5Hz, 1H), 8.01 (d,1H). Exampls 49

BOC-Trp-Lys (epsilon-N-(3-(2-thienyl)acrylyl))-Asp-Phe-NH₂ The tetrapeptide of example 6 (70 mg), active ester of example 48 (31 mg) and N-methylmorpholine (11

mg) were allowed to react under similar conditions to those described in example 8. The product was isolated in a similar manner to yield 47 mg of a white solid.

MS(FAB+) m/e 830 (M+H)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 1.10-1.68 (m, 6H), 1.30 (br s,9H), 2.36-2.68 (m,2H), 2.78-3.18 (m, 6H) , 4.18-4.40 (m, 3H), 4.49 (q, J=7Hz, 1H), 6.38

(d, J=16Hz,1H), 6.82 (d, J=8Hz, 1H), 6.92-7.62 (m, 9H), 7.90-8.02 (m,2H), 8.10 (br t,1H), 8.24 (br d, 1H), 10.83 (br S,1H). Anal calcd for C₄₂H₅₁N₇O₉S H₂O: C 59.49, H 6.30, N 11.56; found: C 59.55, H 6.15, N 11.49.

Example 50

2-Naphthoic acid N-hydroxysuccinimide ester A solution of 2-naphthoic acid (500 mg), N-hydroxysuccinimide (400 mg) and EDCI (610 mg) in methylene chloride (20 mL) was stirred at ambient temperature for 18 h. The product was isolated as described in example 7 to yield 730 mg of a white solid. MS(CI/NH₃) m/e 270 (M+H)⁺. ¹H NMR (CDCl₃, 300MHz) δ 2.94 (br s,4H), 7.55-7.70 (m, 2H), 7.88-8.01 (m, 3H), 8.09 (dd, J=6Hz, 1H), 8.76 (d, J=2Hz, 1H).

Example 51

BOC-Trp-Lys (epsilon-N-(2-naphthoyl))-Asp-Phe-NH₂

The tetrapeptide of example 6 (70 mg), active ester of example 50 (33 mg) and N-methylmorpholine (11 mg) were allowed to react under similar conditions to those described in example 8. The product was purified in an identical manner to yield 52 mg of a white solid.

MS(FAB+) m/e 848 (M+H)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 1.05- 1.68 (m, 6H), 1.29 (br s,9H), 2.42-2.72 (m, 2H), 2.79-3.18

(m, 6H), 4.18-4.42 (m, 3H). 4.52 (q, J=7Hz, 1H), 6.82-7.34

(m, 8H), 7.51-7.63 (m, 2H), 7.84-8.01 (m, 4H), 8.28 (br d,J=7Hz,1H), 8.43 (br s,1H), 8.62 (br t,1H). Anal calcd for C₄₆H₅₃N₇O₉ 0.5H₂O: C 64.47, H 6.35, N 11.44; found:

C 64.67, H 6.27, N 11.42. Example 52

4-Chlorocinnamic acid N-hydroxysuccinimide ester A solution of 4-chlorocinnamic acid (0.08 g), N-hydroxysuccinimide (0.55 g), and EDCI (0.88 g) in

methylene chloride was stirred at ambient temperature for 18 h. The product was isolated as described in example 7 to yield 550 mg of white crystals, mp 192-193 °C.

MS(DCI/NH₃) m/e 297 (M+NH₄)⁺. ¹H NMR(DMSO-d6, 300MHz) δ 2.87 (br s,4H), 7.05 (d, J=17Hz, 1H), 7.56 (d, J=9Hz, 1H), 7.92 (d, 1H), 7.99 (d, 1H).

Example 53

BOC-Trp-Lys (epsilon-N-(4-chlorocinnamoyl))-Asp-Phe-NH₂ The tetrapeptide of example 6 (100 mg), active ester of example 52 (47 mg) and N-methylmorpholine (17 mg) were allowed to react under similar conditions to those described in example 8. The product was isolated in an identical manner to yield 85 mg of a white solid.

MS(FAB+) m/e 858 (M+H)⁺. ¹H NMR(DMSO-d6,300MHz) δ 1.08- 1.65 (m,6H), 1.30 (br s,9H), 2.35-2.62 (m,2H), 2.78-3.20 (m,6H), 4.15-4.40 (m, 3H), 4.48 (q, J=7Hz, 1H), 6.64 (d, J=16Hz,1H), 6.83 (br d, J=7Hz, 1H), 6.91-7.62 (m, 15H), 7.95 (br d, 1H), 8.03 (br s,1H), 8.17 (br S,1H), 8.25 (br d,1H), 8.57 (br d, 1H), 10.87 (br s,1H). Anal calcd for

C₄₄H₅₂C₁N₇O₉ H₂O: C 60.30, H 6.21, N 11.19; found: C 60.65, H 6.18, N 11.06.

Example 54

3-(3,4-Dihydroxyphenyl)propionic acid

N-hydroxysuccinimide ester

A solution of 3-(3,4-dihydroxyphenyl)propionic acid (350 mg), N-hydroxysuccinimide (230 mg) and EDCI (380 mg) in methylene chloride and dimethylformamide was stirred at ambient temperature for 18 h. The solvents were removed in vacuo and the residue was dissolved in ethyl acetate and washed several times with water. The ethyl acetate was dried (MgSO₄) and evaporated to yield 560 mg of a white solid. MS(CI/NH₃) m/e 297 (M+NH₄)⁺ . ¹H NMR(DMSO-d6, 300MHz) δ 2.72-2.90 (m, 8H), 7.95 (br s, 1H), 8.63-8.79

(m,2H).

Example 55

BOC-Trp-Lys (epsilon-N-(3-(3,4-dihydroxyphenyl)- propionyl))-Asp-Phe-NH₂

The tetrapeptide of example 6 (149 mg), active ester of example 54 (60 mg) and N-methylmorpholine (21 mg) were allowed to react under similar conditions to those

described in example 8. The product was isolated in a similar manner to yield 52 mg of a white, flocculent solid. MS(FAB+) m/e 880 (M+Na)⁺. ¹H NMR(DMSO-d6, 300MHz) δ 1.08-1.61 (m,6H), 1.30 (br s, 9H), 2.21-3.16 (m, 12H), 4.22 (br s,2H), 4.33 (m,1H), 4.45 (q, J=7Hz, 1H), 6.39 (br d,1H), 6.56-7.35 (m, 11H), 7.49 (br s,1H), 7.59 (br

d, J=7Hz,1H), 7.77 (m, 1H), 7.95 (br d, 1H), 8.05 (br s,1H), 8.19 (br d,1H) . Anal calcd for C₄₄H₅₅N₇O₁₁ 0.5H₂O

CH₃CO₂H: C 59.60, H 6.52, N 10.58; found: C 59.75, H 6.26, N 10.35.

Example 56

6-Acetoxy-2-naphthoic acid N-hydroxysuccinimide ester A solution of 6-acetoxy-2-naphthoic acid (1.00 g), N-hydroxysuccinimide (0.53 g) and EDCI (0.95 g) in methylene chloride was stirred at ambient temperature for 18 h. The product was isolated as described in example 7 to yield 0.90 g of a white solid. MS(CI/NH₃) m/e 345 (M+NH₄)⁺. ¹H NMR (CDCl₃, 300MHz) δ 2.39 (s,3H), 2.94 (br s,4H), 7.36 (dd, J=9Hz,1H), 7.66 (d, J=2Hz, 1H), 7.91 (d, J=8Hz, 1H), 8.00 (d,1H), 8.10 (dd,1H), 8.75 (br s,1H) .

Example 57

BOC-Trp-Lys (epsilon-N-(6-acetoxy-2-naphthoyl))- Asp-Phe-NH₂

The tetrapeptide of example 6 (145 mg), active ester of example 56 (50 mg) and N-methylmorpholine (16 mg) were allowed to react under similar conditions to those

described in example 8. The product was purified in an identical manner to yield 57 mg of a white solid.

MS(FAB+) m/e 906 (M+H⁾⁺. ¹H NMR(DMSO-d6, 500MHz) δ 1.08-1.68 (m,6H), 1.28 (br s,9H), 2.34 (s,3H), 2.42-2.50

(m,1H), 2.60-2.67 (m, 1H), 2.81-2.95 (m,2H), 3.01-3.14 (m, 2H), 4.18-4.40 (m, 3H), 4.51 (q, J=7Hz, 1H), 6.82 (br d,1H), 6.95 (br t,1H), 7.04 (br t, 1H), 7.10-7.26 (m, 7H), 7.30-7.38 (m,3H), 7.58 (br d, 1H), 7.72 (d, J=2Hz, 1H), 7.88- 7 . 98 (m, 4H) , 8 . 03 (d, J=7Hz , 1H) , 8 . 26 (br d, 1H) , 8 . 47 (br s,1H), 8.62 (br t,1H), 10.80 (br s,1H). Anal calcd for

C₄₈H₅₅N₇O₁₁ H₂O C₅H₅N: C 61.91, H 6.31, N 10.75; found:

C 61.86, H 6.02, N 11.14.

Example 58

alpha-Cyano-3-hydroxycinnamic acid

N-hydroxysuccinimide ester

A solution of a-cyano-3-hydroxycinnamic acid (500 mg), N-hydroxysuccinimide (310 mg) and EDCI (500 mg) in methylene chloride (20 mL) was stirred at ambient

temperature for 18 h. The mixture was washed with water (2X) and brine (2X) then dried over MgSO₄. The solvent was removed in vacuo and the residue chromatographed (hexane/ethyl acetate) to yield 210 mg of a yellow solid. MS(CI/NH₃) m/e 304 (M+NH₄) ⁺. ¹H NMR (CDCl₃, 300MHz) δ 2.91 (br s,4H), 6.99 (d, J=9Hz, 1H), 7.96-8.05 (m, 2H), 8.22 (br S,1H).

Example 59

BOC-Trp-Lys (epsilon-N-(alpha-cyano-3-hydroxycinnamoyl))-

Asp-Phe-NH₂

The tetrapeptide of example 6 (177 mg), active ester of example 58 (70 mg) and N-methylmorpholine (27 mg) were allowed to react under similar conditions to those described in example 8. The product was isolated in a similar manner to yield 42 mg of a white solid. MS (FAB+) m/e 865 (M+H)⁺. ¹H NMR (DMSO-d6, 500MHz) δ 1.00-1.64

(m, 6H), 1.30 (br s,9H), 2.41-2.50 (m, 1H), 2.59-2.66

(m,1H), 2.82-2.95 (m, 2H), 3.02-3.19 (m,4H), 4.18-4.30 (m,2H), 4.47 (br q,1H), 6.81 (br t,1H), 6.88-7.44 (m, 8H), 7.58 (br d,1H), 7.83 (d, J=8Hz, 1H), 7.92 (br d, 1H), 8.02 (s,1H), 8.21-8.29 (m, 2H), 10.79 (br s,1H). Anal calcd for

C₄₅H₅₂N₈O₁₀ H₂O 0.5CH₃CO₂H: C 60.51, H 6.18, N 12.27; found: C 60.47, H 5.92, N 12.27.

Example 60

Cinnamic acid N-hydroxysuccinimide ester A solution of cinnamic acid (1.00 g), N-hydroxysuccinimide (0.80 g) and EDCI (1.30 g) was stirred at ambient temperature for 18 h. The solvent was removed in vacuo and the residue dissolved in ethyl acetate and washed with water and brine. After drying over MgSO₄, the solvent was evaporated to yield 1.20 g of a white solid. MS(CI/NH₃) m/e 263 (M+NH₄)⁺. ¹H NMR(DMSO-d6, 300MHz) δ 2.88 (br s,4H), 7.02 (d, J=16Hz, 1H), 7.45-7.55 (m, 3H), 7.88 (br dd, J=8Hz,2H), 7.99 (d, 1H).

Example 61

BOC-Trp-Lys (epsilon-N-(cinnamoyl))-Asp-Phe-NH₂

The tetrapeptide of example 6 (100 mg), active ester of example 60 (37 mg) and N-methylmorpholine (18 mg) were allowed to react in a similar manner to that described in example 8. The product was isolated in an identical manner to yield 35 mg of a white solid. MS (FAB+) m/e 824 (M+H)⁺. ¹H NMR (DMSO-d6, 500MHz) δ 1.10-1.63 (m, 6H), 1.30 (br s,9H), 2.35-2.61 (m,2H), 2.81-2.96 (m,2H), 3.02-3.18 (m,4H), 4.20-4.30 (m,3H), 4.36 (m, 1H), 4.48 (br q, 1H), 6.64 (d, J=16Hz,1H), 6.79 (br d, 1H), 6.95 (t, J=7Hz, 1H), 7.02-7.60 (m,10H), 7.92 (br s,1H), 8.03 (br s,1H), 8.12 (br s,1H), 8.22 (br d, 1H) , 10.85 (br s,1H). Anal calcd for C₄₄H₅₃N₇O₉ CH₃CO₂H: C 61.50, H 6.57, N 10.91; found: C 61.31, H 6.18, N 11.28.

Example 62

1-Adamantane carboxylic acid N-hydroxysuccinimide aster A solution of 1-adamantane carboxylic acid (800 mg), N-hydroxysuccinimide (580 mg) and EDCI (800 mg) in methylene chloride was stirred at ambient temperature for 18 h. The active ester was isolated as described in example 7 to yield 850 mg of a white solid. MS(CI/NH₃) m/e 278 (M+H)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 1.72 (br t, 6H), 1.97 (br d, 6H), 2.02 (br s, 3H), 2.80 (br s, 4H).

Example 63

BOC-Trp-Lys (epsilon-N-(1-adamantoyl))-Asp-Phe-NH₂

The tetrapeptide of example 6 (100 mg), active ester of example 62 (48 mg) and N-methylmorpholine (16 mg) were allowed to react under similar conditions to those described in example 8. The product was isolated in an identical manner to yield 105 mg of a white solid. MS(FAB+) m/e 856 (M+H) ⁺ . ¹H NMR (DMSO-d6, 300MHz) δ 1.11-1.54 (m, 6H), 1.30 (br s,9H), 1.62 (br s,6H), 1.75 (br d, 6H), 4.15-4.29 (m,2H), 4.30-4.39 (m,1H), 4.42-4.51 (m, 1H), 6.77 (d, J=7Hz, 1H), 6.92-7.48 (m,10H), 7.58 (br d, J=8Hz, 1H), 7.89-8.08 (m, 3H), 8.21 (br d, 1H), 10.85 (br s,1H). Anal calcd for C₄₆H₆₁N₇O₉

1.5H₂O: C 62.56,H 7.25, N 11.10; found: C 62.39, H 7.01, N 11.04.

Example 64

1-Adamantane acetic acid N-hydroxysuccinimide ester

A solution of 1-adamantane acetic acid (800 mg), N-hydroxysuccinimide (530 mg) and EDCI (850 mg) in methylene chloride (60 mL) was allowed to stir at ambient temperature for 18 h. The product was isolated as described in example 7 to yield 860 mg of a white solid. MS(DCI/NH₃) m/e 292 (M+H)⁺. ¹H NMR (CDCl₃,300MHz) d 1.61-1.76 (m, 12H), 2.01 (br s,3H), 2.34 (s,2H), 2.83 (br s,4H).

Example 65

BOC-Trp-Lys (epsilon-N- (1-adamantane acetyl))-Asp-Phe-NH₂ The tetrapeptide of example 6 (100 mg), active ester of example 64 (50 mg) and N-methylmorpholine (16 mg) were allowed to react under similar conditions to those described in example 8. The product was isolated in a similar manner to yield 68 mg of a white solid. MS (FAB+) m/e 870 (M+H)⁺. ¹H NMR(DMSO-d6, 300MHz) δ 1.10-1.69 (m, 18H), 1.30 (br s, 9H), 1.80 (s,2H), 1.90 (br s,3H), 2.25-2.72 (m,2H), 2.78-3.25 (m, 6H), 4.15-4.30 (m,2H), 4.32-4.42 (m, 1H), 4.51

(q, J=8Hz,1H), 6.85 (d, J=8Hz, 1H), 6.92-7.35 (m, 9H), 7.55-7.68 (m,2H), 8.00 (br t,1H), 8.28 (br d, 1H), 10.80 (br s,1H). Anal calcd for C₄₇H₆₃N₇O₉ H₂O: C 63.56, H 7.37, N 11.03; found: C 63.43, H 7.12, N 10.73.

Example 66

4-Methoxycinnamic acid N-hydroxysuccinimide ester

A solution of 4-methoxycinnamic acid (0.05 g), N- hydroxysuccinimide (0.39 g) and EDCI (0.59 g) in methylene chloride (20 mL) was stirred at ambient temperature for 18 h. The ester was isolated in a similar manner as described in example 7 to yield 0.66 g of a white solid. MS(CI/NH₃) m/e 275 M⁺ . 1H NMR (CDCl₃, 300MHz) δ 2.88 (br s, 4H), 3.86 (s,3H), 6.45 (d, J=16Hz,1H), 6.93 (dt, J=9Hz, 2H), 7.52

(dt,2H), 7.88 (d,1H).

Example 67

BOC-Trp-Lys (epsilon-N-(4-methoxycinnamoyl))-Asp-Phe-NH₂ The tetrapeptide of example 6 (71 mg), active ester of example 66 (35 mg) and N-methylmorpholine (11 mg) were allowed to react under similar conditions to those

described in example 8. The peptide was isolated in an identical manner to yield 45 mg of a white solid.

MS(FAB+) m/e 854 (M+H)⁺ . ¹H NMR(DMSO-d6, 300MHz) δ 1.08-1.64 (m,6H), 1.30 (br s,9H), 2.35-2.65 (m,2H), 2.78-3.23 (m,6H), 4.16-4.40 (m, 3H), 4.49 (q, J=7Hz, 1H), 6.49

(d, J=16Hz,1H), 6.82 (br d, J=8Hz, 1H), 6.91-7.53 (br d, 1H), 7.59 (d, J=7Hz,1H), 7.89-8.09 (m, 3H), 8.25 (br d, 1H). Anal calcd for C₄₅H₅₅N₇O₁₀ 2H₂O: C 60.73, H 6.68, N 11.02;

found: C 60.61, H 6.35, N 10.97.

Example 68

4-(Dimethylamino)cinnamic acid N-hydroxysuccinimide ester A solution of 4-(dimethylamino)cinnamic acid (0.50 g), N-hydroxysuccinimide (0.37 g) and EDCI (0.56 g) in methylene chloride (20 mL) and dimethylformamide (5 mL) was stirred at ambient temperature for 18 h. The product was isolated as described in example 7 to yield 0.15 g of a yellow solid. MS(CI/NH3) m/e 289 (M+H)⁺. ¹H

NMR (CDCl₃, 300MHz) δ 2.86 (br s, 4H), 3.05 (s,6H), 6.32 (d, J=16Hz,1H), 6.67 (dt, J=9Hz, 2H), 7.46 (dt,2H), 7.83 (d,1H). Example 69

BOC-Trp-Lys (epsilon-N-(4-(dimethylamino)cinnamoyl))- Asp-Phe-NH₂

The tetrapeptide of example 6 (70 mg), active ester of example 68 (37 mg) and N-methylmorpholine (11 mg) were allowed to react under similar conditions to those described in example 8. The peptide was isolated in an identical manner to yield 31 mg of a white solid.

MS(FAB+) m/e 867 (M+H)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 1.06-1.68 (m,6H), 1.30 (br s,9H), 2.41-2.71 (m,2H), 2.78-3.25 (m, 6H), 4.16-4.31 (m, 2H) , 4.32-4.41 (m, 1H) , 4.51

(q, J=7Hz,1H), 6.25 (d, J=16Hz, 1H), 6.68 (d, J=9Hz, 2H), 6.82-7.48 (m, 11H), 7.59 (d, J=8Hz, 1H), 7.82-7.97 (m, 3H), 8.28 (br d,1H). Anal calcd for C₄₆H₅₈N₈O₉ 0.5H₂O: C 63.07,H 6.79, N 12.79; found: C 62.97, H 6.68, N 12.69.

Example 70

4-Bromocinnamic acid N-hydroxysuccinimide ester

A solution of 4-bromocinnamic acid (0.50 g), N-hydroxysuccinimide (0.30 g) and EDCI (0.46 g) in methylene chloride (20 mL) was stirred at ambient temperature for 18 h. The active ester was isolated as described in example 7 to yield 0.65 g of a white solid. MS(CI/NH₃) m/e 341 (M+H)⁺. ¹H NMR (CDCl₃,300MHz) δ 2.88 (br s,4H), 6.59 (d, J=17Hz,1H), 7.43 (dt, J=9Hz, 2H), 7.57 (d,2H), 7.85 (d, 1H).

Example 71

BOC-Trp-Lys (epsilon-N-(4-bromocinnamoyl))-Asp-Phe-NH₂ The tetrapeptide of example 6 (70 mg), active ester of example 70 (40 mg) and N-methylmorpholine (11 mg) were allowed to react under similar conditions to those described in example 8. The final product was isolated in a similar manner to yield 40 mg of a white solid.

MS(FAB+) m/e 904 (M+H)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 1.08-1.65 (m,6H), 1.29 (br s, 9H), 2.40-2.70 (m,2H), 2.77-3.32 (m,6H), 4.15-4.31 (m, 2H), 4.32-4.42 (m, 1H) , 4.50

(q, J=7Hz, 1H), 6.63 (d, J=16Hz, 1H), 6.82-7.62 (m, 14H), 7.92 (br t,2H), 8.13 (t,1H), 8.28 (br d, 1H). Anal calcd for

C₄₄H₅₂N₇O₉Br H₂O CH₃CO₂H: C 56.33, H 5.96, N 10.00;

found: C 56.00, H 5.57, N 10.19.

Example 72

6-Hydroxy-2-naphthoic acid N-hydroxysuccinimide ester A solution of 6-hydroxy-2-naphthoic acid (1.0 g), N-hydroxysuccinimide (0.65 g) and EDCI (1.1 g) in methylene chloride was stirred at ambient temperature. The product was isolated as described in example 7 to yield 0.32 g of a white solid. MS(CI/NH₃) m/e 303 (M+NH₃)⁺. ¹H NMR(DMSO-d6, 300MHz) δ 2.92 (br s,4H), 7.22-7.29 (m,2H), 7.91

(s,2H), 8.10 (d, J=8Hz,1H), 8.71 (s,1H), 10.49 (s,1H).

Example 73

BOC-Trp-Lys (epsilon-N-(6-hydroxy-2-naphthoyl))- Asp-Phe-NH₂

The tetrapeptide of example 6 (100 mg), active ester of example 72 (40 mg) and N-methylmorpholine (37 mg) were reacted under similar conditions to those described in example 8. The product was isolated in an identical

manner to yield 38 mg of a white solid. MS(FAB+) m/e 864

(M+H)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 1.08-1.68 (m, 6H), 1.29

(br s,9H), 2.36-2.65 (m,2H), 2.77-3.19 (m, 6H), 4.18-4.40

(m, 3H), 4.49 (br q, 1H), 6.77-8.51 (m,20H). Anal calcd for C₄₆H₅₃F₇O₁₀ 1.5H20: C 62.01, H 6.34, N 11.01; found: C

61.71, H 6.04, N 10.92.

Example 74

2,4-Dichlorocinnamic acid N-hydroxysuccinimide ester A solution of 2,4-dichlorocinnamic acid (1.0 g), N-hydroxysuccinimide (0.6 g) and EDCI (0.9 g) in methylene chloride was stirred at ambient temperature for 18 h. The product was isolated as described in example 7 to yield 0.9 g of a white solid. MS(CI/NH₃) m/e 331 (M+NH₃)⁺. ¹H

NMR (CDCl₃, 300MHz) δ 2.89 (br s,4H), 6.60 (d, J=16Hz, 1H), 7.32 (dd, J=8Hz,1H), 7.48 (d, J=2Hz, 1H), 7.62 (d, 1H), 8.26 (d, 1H).

Example 75

BOC-Trp-Lys (epsilon-N-(2,4-dichlorocinnamoyl))-Asp-Phe-NH₂ The tetrapeptide of example 6 (100 mg), active ester of example 74 (46 mg) and N-methylmorpholine (20 mg) were allowed to react under similar conditions to those described in example 8. The product was isolated in an identical manner to yield 45 mg of a white solid. MS (FAB+) m/e 892 M+. ¹H NMR(DMSO-d6, 500MHz) δ 1.09-1.65 (m, 6H), 1.29 (br s,9H), 2.42-2.68 (m, 2H), 2.80-2.94 (m,2H), 3.00-3.20

(m,4H), 4.18-4.30 (m, 2H), 4.38 (m, 1H), 4.50 (br q, 1H), 6.68 (d,J=16,1H), 6.82 (d, J=8Hz,1H), 6.92-7.44 (m, 10H), 7.55-7.69 (m,3H), 7.84-7.92 (m, 2H), 8.18-8.28 (m, 2H). Anal calcd for C₄₄H₅₁Cl₂N₇O₉ 1.5H₂O: C 57.45, H 5.92, N 10.66; found: C 57.90, H 5.78, N 10.59.

Example 76

4-Nitrocinnamic acid N-hydroxysuccinimide ester

A solution of 4-nitrocinnamic acid (1.0 g), N-hydroxysuccinimide (0.6 g) and EDCI (1.0 g) in methylene chloride was stirred at ambient temperature for 18 h. The product was isolated as described in example 7 to yield 1.09 g of a white solid. MS(CI/NH₃) m/e 308 (M+NH₄)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 2.88 (br s,4H), 7.25 (dd, J=16Hz, 1H), 8.08-8.18 (m,3H), 8.28-8.33 (m, 2H).

Example 77

BOC-Trp-Lys (epsilon-N-(4-nitrocinnamoyl))-Asp-Phe-NH₂ The tetrapeptide of example 6 (100 mg), active ester of example 76 (42 mg) and N-methylmorpholine (28 mg) were allowed to react under similar conditions to those

described in example 8 and purified in an identical manner to yield 38 mg of a white solid. MS(FAB+) m/e 891

(M+Na)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 1.09-1.65 (m, 6H), 1.29 (br s,9H), 2.36-2.65 (m,2H), 2.79-3.22 (m, 6H), 4.17-4.30 (m,2H), 4.35 (m,1H), 4.48 (br q, 1H), 6.76-6.85 (m,2H), 6.94 (br t, J=5Hz,1H), 7.03 (t,1H), 7.08-7.60 (m, 10H), 7.76 (d,J=6H,2H), 7.88-8.04 (m,2H), 8.18-8.32 (m, 3H). Anal calcd for C₄₄H₅₂N₈O₁₁ H₂O: C 58.34, H 6.17, N 11.85;

found: C 58.11, H 5.88, N 12.25.

Example 78

3,4-Dimethoxycinnamic acid N-hydroxysuccinimide ester A solution of 3, 4-dimethoxycinnamic acid (1.0 g), N-hydroxysuccinimide (0.56 g) and EDCI (0.95 g) in methylene chloride was stirred at ambient temperature. The active ester was isolated as described in example 7 to yield 0.81 g of a white solid. ¹H NMR(DMSO-d6, 300MHz) δ 2.85 (br s,4H), 3.82 (s,6H), 6.89 (d, J=16Hz, 1H), 7.04 (d, J=9Hz, 1H), 7.40 (dd, 1H), 7.51 (d, J=2Hz, 1H), 7.89 (d,1H).

Example 79

BOC-Trp-Lys (epsilon-N-(3,4-dimethoxvcinnamovl))-

Asp-Phe-NH₂

The tetrapeptide of example 6 (100 mg), active ester of example 78 (45 mg) and N-methylmorpholine (28 mg) were allowed to react under similar conditions to those

described in example 8. The final product was purified in an identical manner to yield 32 mg of a white solid.

MS(FAB+) m/e 884 (M+H)⁺. ¹H NMR(DMSO-d6, 300MHz) δ 1.06-1.69 (m,6H), 1.29 (br s,9H), 2.38-2.52 (m, 1H), 2.60-2.71 (m,1H), 2.80-3.25 (m, 6H) , 3.78 (br s,6H), 4.17-4.43

(m,3H), 4.51 (br q, 1H), 6.51 (d,J=16Hz, 1H), 6.82-7.40 (m, 9H), 7.60 (d, J=8Hz, 1H), 7.88-8.06 (m,3H), 8.29 (d, 1H). Anal calcd for C₄₆H₅₇N₇O₁₁ H₂O: C 61.18, H 6.59, N 10.86; found: C 60.85, H 6.38, N 10.68.

Example 80

4-(3-Ouinolyl)-3-butenoic acid

N-hydroxysuccinimide ester

A solution of 4-(3-quinolyl)-3-butenoic acid (300 mg), N-hydroxysuccinimide (175 mg) and EDCI (310 mg) in methylene chloride was stirred at ambient temperature. The product was isolated as described in example 7 to yield 280 mg of a light yellow solid. MS(CI/NH₃) m/e 311 (M+H)⁺. ¹H NMR (CDCl₃, 300MHz) δ 2.86 (br s,4H), 3.64 (dd, J=7Hz, 2H), 6.51 (dt, J=16Hz, 1H), 6.81 (d, 1H), 7.54 (br t,J=7Hz, 1H), 7.69 (br t,1H), 7.82 (br d, J=8Hz, 1H), 8.08 (br q, 2H), 8.99 (d, J=2Hz, 1H) .

Example 81

BOC-Trp-Lys (epsilon-N-(4-(3-quinolyl)-3-butenoyl))- Asp-Phe-NH₂

The tetrapeptide of example 6 (120 mg), active ester of example 80 (55 mg) and N-methylmorpholine (19 mg) were reacted under similar conditions to those described in example 8. The product was isolated under identical conditions to yield 32 mg of a white solid. MS(FAB+) m/e 889 (M+H)⁺. ¹H NMR (DMSO-d6, 500MHz) δ 1.10-1.63 (m, 6H), 1.29 (br s,9H), 2.41-2.48 (m, 1H), 2.58-2.66 (m, 1H), 2.82-3.16 (m,8H), 4.19-4.32 (m, 2H), 4.37 (m, 1H), 4.50 (q, 1H), 6.65 (br t,1H), 6.80 (d, J=8Hz, 1H), 6.96 (t, J=6Hz, 1H), 7.03 (t,1H), 7.10-7.41 (m, 9H), 7.58 (m, 2H), 7.71 (br t,1H), 7.88-8.00 (m,4H), 8.22 (d, 1H), 8.29 (d, 1H), 9.01

(d, J=3Hz, 1H). Anal calcd for C₄₈H₅₆N₈O₉ H₂O ·75CH₃COOH: C 62.38, H 6.46, N 11.73; found: C 62.27, H 6.26, N

11.79.

Example 82

BOC-Trp-Lys (epsilon-N-(3,4-dihydroxycinnamoyl))- Asp-Phe-NH₂

A solution of 3, 4-dihydroxycinnammic acid (80 mg), N-hydroxysuccinimide (50 mg) and 1,3-dicyclohexylcarbodiimide (95 mg) in DMF was stirred at ambient temperature for 18 h. The reaction mixture was cooled to 0°C and the tetrapeptide of example 6 (320 mg) and N-methylmorpholine (92 mg) were added and

allowed to react for 24 h. The product was isolated in a similar manner as described for example 8 to yield a white solid. MS(FAB+) m/e 856 (M+H)⁺. ¹H NMR(DMSO-d6, 500MHz) δ 1.06-1.66 (m,6H), 1.29 (br s,9H), 2.40-2.55 (m,2H), 2.81-3.17 (m,6H), 4.17-4.32 (m, 2H) , 4.33-4.40 (m, 1H) , 4.50

(q, 1H), 6.13 (d,J=16Hz,1H), 6.42-6.54 (m, 1H), 6.72

(d, J=7Hz,1H), 6.77-6.85 (m,2H), 6.92-7.42 (m, 12H), 7.58 (d, 1H), 7.88-7.97 (m,2H), 8.23 (d, 1H). Anal calcd for C₄₄H₅₃N₇O_l1 H₂O CH₃COOH: C 59.15, H 6.37, N 10.50; found: C 59.44, H 6.21, N 10.52.

Example S3

3,4-Dichlorocinnamic acid N-hydroxysuccinimide ester A solution of 3,4-dichlorocinnamic acid (1.0 g), N-hydroxysuccinimide (0.6 g) and EDCI (0.9 g) in methylene chloride was stirred at ambient temperature. The solvent was removed in vacuo and the residue was dissolved in ethyl acetate and washed several times with water. After drying

(MgSO₄), the ethyl acetate was removed in vacuo and the resulting solid was recrystallized (ethyl acetate) to yield 1.19 g of a white solid, mp 192-196 °C. MS(CI/NH₃) m/e 314

(M+H)⁺. ¹H NMR (CD3OD,300MHz) δ 2.87 (s,4H), 6.86

(d, J=17Hz,1H), 7.61 (d, J=8Hz,1H), 7.68 (dd, J=2Hz, 1H), 7.91

(d, 1H), 7.94 (d,1H).

Example S4

BOC-Trp-Lys (epsilon-N-(3,4-dichlorocinnamoyl))-Asp-Phe-NH₂ The tetrapeptide of example 6 (100mg), active ester from example 83 (43 mg) and N-methylmorpholine (15 mg) were allowed to react under similar conditions to

those described in example 8. The final product was isolated as described to yield 62 mg of a white solid.

MS(FAB+) m/e 892 (M+H)⁺. ¹Η NMR (DMSO-d6, 500MHz) δ 1.06-1.66 (m, 6H), 1.29 (br s, 9H), 2.35-2.55 (m,2H), 2.81-3.18 (m, 6H) , 4.19-4.32 (m, 2H), 4.36 (m, 1H), 4.47 (q, 1H), 6.72 (d, J=16Hz,1H), 6.78 (br d, 1H), 6.95 (br t, 1H), 7.02-7.65 (m, 12 H), 7.79 (d, J=3Hz, 1H), 7.91 (br d, 1H), 8.04 (br s,1H), 8.14-8.24 (m,2H). Anal calcd for C₄₄H₅₁Cl₂N₇O₉ CH₃COOH 1.75H₂O: C 56.18, H 5.98, N 9.97; found: C

55.94, H 5.59, N 10.23.

Example 85

BOC-D-Trp-Lys (epsilon-N-Cbz)-ASP(beta-Bn)-Phe-NH₂

The compound was prepared in a similar manner to example 5 via coupling of BOC-D-Trp with tripeptide of example 4 using EDCI and 1-hydroxybenzotriazole.

Example 86

BOC-D-Trp-Lys-Asp-Phe-NH₂

The tetrapeptide was prepared from example 85 in an identical manner to example 6.

Example 87

BOC-D-Trp-Lys (epsilon-N-(3-(4-hydroxyphenyl)propionyl))- Asp-Phe-NH₂

The tetrapeptide of example 86 (60 mg), N-methylmorpholine (9 mg) and 3-(4-hydroxyphenyl)propionic acid N-hydroxysuccinimide ester (28 mg) in DMF (6 mL) were allowed to react under similar conditions to those

described in example 8. The product was isolated in an identical manner to yield 54 mg of a white solid.

MS(FAB+) m/e 842 (M+H)⁺. ¹H

NMR (DMSO-d6, 300MHz) δ 0.98-1.58 (m, 6H), 1.30 (br s,9H), 2.22-3.15 (m, 12H), 4.09-4.38 (m, 3H), 4.43-4.55 (m, 1H), 6.63 (dt, J=8Hz,1H), 6.85-7.44 (m, 12H), 7.58 (br d,J=7Hz,1H), 7.74 (br t,1H), 7.91 (br d, 1H), 8.00 (br d, J=7Hz, 1H), 8.14 (br d, J=7Hz,1H). Anal calcd for C₄4H₅₅N₇O₁₀ 0.5CH₃COOH: C 59.52, H 6.77, N 10.80; found: C 59.88, H 6.19, N 11.05.

Example 88

BOC-D-Trp-Lys (epsilon-N-(4-hydroxycinnamoyl))-Asp-Phe-NH₂ The tetrapeptide of example 87 (75 mg), active ester of example 15 (34 mg) and N-methylmorpholine (12 mg) were allowed to react under similar conditions to those described in example 8. The product was isolated as described to yield 63 mg of a white powder. MS (FAB+) m/e 840 (M+H)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 1.01-1.62 (m, 6H), 1.30 (br s,9H), 2.37-3.13 (m,8H), 4.10-4.39 (m,3H), 4.50 (br q,J=7Hz, 1H), 6.40 (d, J=16Hz,1H), 6.77 (d, J=9Hz,1H), 6.86-7.40 (m, 10H), 7.57 (br d,1H), 7.85 (br d, 1H) , 7.92-8.05 (m, 2H) , 8.15 (br d,1H). Anal calcd for C₄₄H₅₃N₇O₁₀ CH₃COOH 1.5H₂O: C 59.60, H 6.52, N 10.58; found: C 59.42, H 5.93, N 10.93. Example 89

BOC-alpha-Nal-Lys (epsilon-N-Cbz)-Asp(beta-Bn)-Phe-NH₂ The compound was prepared in a similar manner to example 5 via coupling of BOC-alpha-Nal with tripeptide of example 4 using EDCI and 1-hydroxybenzotriazole.

Example 90

BOC-alpha-Nal-Lys-Asp-Phe-NH₂

The tetrapeptide was prepared from example 89 in an identical manner to example 6.

Example 91

BOC-alpha-Nal-Lys (epsilon-N-(3-(4-hydroxyphenyl)

propionyl))-Asp-Phe-NH₂

The tetrapeptide of example 90 (55 mg), 3-(4-hydroxyphenyl)propionic acid N-hydroxysuccinimide ester

(25 mg) and N-methylmorpholine (9 mg) were allowed to react under similar conditions to those described in example 8. The product was isolated in an identical manner to yield 46 mg of a white solid. MS (FAB+) m/e 853

(M+H)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 1.12-1.62 (m, 6H), 1.23

(br s,9H), 2.28 (t, J=8Hz, 2H), 2.34-2.62 (m,2H), 2.67

(t,2H), 2.78-3.26 (m, 6H), 4.22-4.40 (m,3H), 4.48 (br q,1H), 6.63 (dt, J=8Hz, 2H), 6.94 (dt,2H), 7.07-7.28 (m, 7H),

7.38-7.60 (m,5H), 7.78 (br s,2H), 7.91 (br d, 2H), 8.02 (br d, 1H), 8.15 (d,1H), 8.30 (d,1H). Anal calcd for

C₄₆H₅₆N₆O₁₀ 2H₂O: C 62.15, H 6.80, N 9.45; found: C

62.26, H 6.46, N 9.38.

Example 92

BOC-alpha-Nal-Lys (epsilon-N-(4-hydroxycinnamoyl))- Asp-Phe-NH₂

The tetrapeptide of example 90 (56 mg), active ester of example 15 (25 mg) and N-methylmorpholine (9 mg) were allowed to react under similar conditions to those described in example 8. The final product was isolated in an

identical manner to yield 41 mg of a white, flocculent solid. MS(FAB+) m/e 873 (M+Na)⁺. ¹H NMR(DMSO-d6, 300MHz) δ 1.11-1.68 (m, 6H), 1.23 (br s,9H), 2.38-2.69 (m,2H), 2.78-3.26 (m,6H), 4.25-4.41 (m, 3H), 4.49 (br q, 1H), 6.40

(d, J=16Hz,1H), 6.75 (d, J=8Hz, 2H), 7.07-7.62 (m, 13H), 7.78 (br t,2H), 7.88-8.06 (m, 4H), 8.16 (br d, 1H), 8.32 (br d, 1H). Anal calcd for C₄₆H₅₄N₆O₁₀ 1.5H₂O: C 62.93, H 6.54, N 9.57; found: C 62.73, H 6.27, N 9.42.

Example 93

BOC-beta-Nal-Lys (epsilon-N-Cbz)-Asp(beta-Bn)-Phe-NH₂ The tetrapeptide was prepared by coupling BOC-beta-Nal with tripeptide of example 4 using EDCI and 1-hydroxybenzotriazole in a manner similar to that described in Example 5.

Example 94

BOC-beta-Nal-Lys-Asp-Phe-NH₂

The tetrapeptide was prepared from the resultant compound of example 93 under identical conditions to those described in example 6.

Example 95

BOC-beta-Nal-Lys (epsilon-N-(3-(4-hydroxyphenyl)- propionyl))-Asp-Phe-NH₂

The tetrapeptide of example 94 (55 mg), 3-(4-hydroxyphenyl)propionic acid N-hydroxysuccinimide ester (25 mg) and N-methylmorpholine (9 mg) were allowed to react under similar conditions to those described for example 8. The product was purified in a similar manner to yield 56 mg of a white solid. MS(FAB+) m/e 853 (M+H)⁺. ¹H NMR(DMSO-d6, 300MHz) δ 1.11-1.72 (m, 6H), 1.22 (br s, 9H), 2.28 (br t,J=8Hz,2H), 2.32-2.60 (m, 2H), 2.68 (t,2H), 2.78-3.29

(m, 6H), 4.18-4.38 (m, 3H) , 4.44 (br q, 1H), 6.62 (d, J=9Hz, 2H), 6.95 (d,2H), 7.01 (d, J=9Hz, 1H), 7.09 (br s,2H), 7.12-7.28 (m,10H), 7.40-7.53 (m, 4H), 7.71-7.89 (m, 5H), 7.98-8.12

(m,2H), 8.25 (d,1H). Anal calcd for C₄₆H₅₆N₆O₁₀ H₂O

1.25CH₃COOH: C 61.57, H 6.71, N 8.88; found: C 61.45, H 6.34, N 9.28.

Example ?6

BOC-beta-Nal-Lys (epsilon-N-(4-hydroxycinnamoyl))- Asp-Phe-NH₂

The tetrapeptide of example 94 (56 mg), active ester of example 15 (25 mg) and N-methylmorpholine (9 mg) were allowed to react under similar conditions to those

described for example 8. The product was isolated in an identical manner to yield 44 mg of a white solid.

MS(FAB+) m/e 851 (M+H)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 1.11-1.72 (m,6H), 1.22 (br s,9H), 2.34-2.65 (m,2H), 2.79-2.96 (m,2H), 3.01-3.29 (m, 4H), 4.20-4.41 (m, 3H), 4.48 (br

q, 1H), 6.42 (d, J=16Hz, 1H), 6.76 (d, J=8Hz, 2H), 7.02

(d, J=8Hz,1H), 7.10-7.54 (m,l1H), 7.71-7.89 (m, 4H), 7.96- 8.06 (m,2H), 8.28 (br d, 1H). Anal calcd for C₄₆H₅₄N₆O₁₀

2H₂O: C 62.29, H 6.59, N 9.47; found: C 62.36, H 6.21, N 9.34

Example 97

CBZ-(NMe)Phe-NH₂

To a -10°C solution of CBZ-(NMe)Phe (20.6 g) and N-methylmorpholine (7.01 g) in tetrahydrofuran (500 mL) was added isobutylchloroformate (9.5 g). After stirring for 5 min, a solution of aqueous ammonium hydroxide (12 mL) was added. After stirring an additional 15 min at -10 °C, the reaction was allowed to warm to ambient temperature. The mixture was quenched with the addition of water and the product was collected and dried to yield 18 g of a white solid. MS(CI/NH₃) m/e 313 (M+H)⁺. ¹H NMR(DMSO-d6, 300MHz) δ 2.82-2.97 (m, 1H) , 3.13-3.28 (m, 1H) , 3.35 (s,3H), 4.75-5.05 (m,3H), 7.13-7.55 (m, 10H).

Example 98

(NMe)Phe-NH₂

The product was prepared from example 97 using the identical conditions described in example 6.

Example 99

BOC-Asp(beta-Bn)-(NMe)Phe-NH₂

To a 0°C solution of BOC-(beta-Bn)Asp (39.9 g) in methylene chloride (350 mL) was added EDCI (11.6 g).

After stirring 1 h, the amide of example 98 (8.33 g) was added to the reaction and allowed to stand at ambient temperature for 18 h. The solvent was removed in vacuo, the residue dissolved in ethyl acetate and washed with 1M H₃PO₄ then with saturated sodium bicarbonate solution. After drying (MgSO₄), the ethyl acetate was evaporated in vacuo to yield 25.4 g of a white solid.

Example 100

Asp(beta-Bn)-(NMe)Phe-NH₂ Hydrochloride The dipeptide was prepared from example 99 employing similar reaction conditions to those described in example 2.

Example 101

BOC-Lys (epsilon-N-Cbz)-ASP(beta-Bn)-(NMe)Phe-NH₂

The coupling of BOC-Lys (epsilon-N-Cbz) and dipeptide of example 100 was conducted under similar conditions to those described in example 3.

Example 102

Lys (epsilon-N-Cbz)-ASP(beta-Bn)-(NMe)Phe-NH₂

Hydrochloride

The tripeptide was prepared from example 101 in a similar manner to that described for example 4.

Example 103

BOC-Trp-Lys (epsilon-N-Cbz)-ASP(beta-Bn)-(N- Me)Phe-NH₂

The coupling of BOC-Trp with the tripeptide of example 102 was performed under identical conditions to those described for example 5.

Example 104

BOC-Trp-Lys-Asp-(NMe)Phe-NH₂

The tetrapeptide was prepared from example 103 employing the procedure described for example 6.

Example 105

BOC-Trp-Lys (epsilon-N-(3-(4-hydroxyphenyl)propionyl))- Asp-(NMe)Phe-NH₂

The tetrapeptide of example 104 (43 mg), N-methylmorpholine (8 uL) and 3-(4-hydroxyphenyl)propionic acid N-hydroxysuccinimide ester (20 mg) were allowed to react under similar conditions to those described in example 8. The product was isolated in an identical manner to yield 26 mg of a white solid. MS (FAB+) m/e 856 (M+H)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 1.08-1.61 (m, 6H), 1.29 (br s,9H), 2.21-2.34 (m,4H), 2.62-3.25 (m, 11H) , 4.12-5.27 (m,4H), 6.62 (d, J=8Hz,2H), 6.74-7.84 (m, 13H), 8.18 (br d, 1H), 8.58 (br d, 2H). Anal calcd for C₄₅H₅₇N₇O₁₀ CH₃COOH 1.5H₂O: C 59.86, H 6.84, N 10.40; found: C 59.54, H 6.52, N 10.32.

Example 106

BOC-Trp-Lys (epsilon-N-(4-hydroxycinnamoyl))-Asp-(N- Me)Phe-NH₂

The tetrapeptide of example 104 (107 mg), active ester of example 15 (48 mg) and N-methylmorpholine (20 uL) were allowed to react under similar conditions to those described in example 8. The product was isolated in an identical manner to yield 48 mg of a white solid.

MS(FAB+) m/e 854 (M+H)⁺. ¹H NMR(DMSO-d6, 300MHz) δ 1.05-1.62 (m,6H), 1.29 (br s, 9H), 2.26-2.54 (m,2H), 2.69-3.26 (m, 9H), 4.15-5.20 (m, 4H), 6.42 (d, J=16Hz, 1H), 6.72-6.85

(m,3H), 6.90-7.65 (m, 14H) , 7.79-8.00 (m,2H), 8.24 (br d,1H), 8.59 (m, 1H) , 9.82 (br s,1H). Anal calcd for

C₄₅R₅₆N₇O₁₀ CH₃COOH 0.5H₂O: C 61.09, H 6.65 N 10.61;

found: C 60.66 H, 6.26, N 11.00.

Example 107

BOC-Trp-Lys (epsilon-N-(4-chlorocinnamoyl))-Asp-(N

Me)Phe-NH₂

The tetrapeptide of example 104 (113 mg), active ester of example 52 (50 mg) and N-methylmorpholine (18 uL) were allowed to react under similar conditions to those described in example 8. The product was purified

employing identical conditions to yield 44 mg of a white powder. MS (FAB+) m/e 872 (m+H)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 1.06-1.51 (m,6H), 1.28 (br s,9H), 2.22-2.51 (m,2H), 2.69-3.31 (m,9H), 4.13-5.18 (m, 4H), 6.55-8.31 (m, 18H). Anal calcd for C₄₅H₅₇ClN₇O₉ 1.5H₂O: C 60.02, H.6.49, N 10.89; found: C 60.16,H 6.14, N 10.88.

Example 108

BOC-Trp-Lys (epsilon-N-(6-hydroxy-2-naphthoyl))- ASP-(NMe)Phe-NH₂

The tetrapeptide of example 104 (156 mg), active ester of example 72 (64 mg) and N-methylmorpholine (24 uL) were allowed to react under similar conditions to those described for example 8. The product was isolated as described to yield 78 mg of a white solid. MS(FAB+) m/e 878 (M+H)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 1.04-1.65 (m, 6H), 1.28 (br d,9H), 2.25-2.59 (m,2H), 2.69-3.35 (m, 9H) , 4.15-5.15 (m,4H), 6.81 (br t,1H), 6.91-7.98 (m, 12H), 8.22-8.62 (m,4H). Anal calcd for C₄₇H₅₅N₇O₁₀ H₂O: C 63.00, H 6.41, N 10.94; found: C 63.27, H 6.38, N 10.66.

Example 109

BOC-Trp-Lys (epsilon-N-(6-acetoxy-2-naphthoyl))- Asp-(NMe)Phe-NH₂

The tetrapeptide of example 104 (105 mg), active ester from example 56 (54 mg) and N-methylmorpholine (17 uL) were allowed to react under similar conditions employed of example 8. The product was purified as described to yield 59 mg of a flocculent solid. MS (FAB+) m/e 920 (M+H)⁺. ¹H NMR(DMSO-d6, 300MHz) δ 1.06-1.65

(m,6H), 1.28 (br d,9H), 2.34 (s,3H), 2.68-3.95 (m, 9H), 4.14-5.18 (m,4H), 6.81 (br t,1H), 6.91-8.07 (m, 12H), 8.26 (br d, 1H), 8.48 (br d, 1H) , 8.55-8.70 (m, 3H). Anal calcd for C₄₉H₅₇N₇O₁₁ 1.5H₂O: C 62.14, H 6.39, N 10.38; found: C 61.94, H 6.13, N 10.29.

Example 110

BOC-D-Trp-Lys (epsilon-N-Cbz)-ASP(beta-Bn)-(N

Me)Phe-NH₂

The tetrapeptide was prepared via coupling of BOC-D-Trp with tripeptide of example 102 employing similar conditions to those described for example 5.

Example 111

BOC-P-Trp-Lys-Asp-(NMe)Phe-NH₂

The above peptide was prepared from example 110 in a similar manner to that described for example 6.

Example 112

BOC-D-Trp-Lys (epsilon-N-(3-(4-hydroxyphenyl)propionyl))- Asp-(NMe)Phe-NH₂

The tetrapeptide of example 111 (60 mg), N-methylmorpholine (9 mg) and 3-(4-hydroxyphenyl)propionic acid N-hydroxysuccinimide ester (28 mg) were allowed to react under similar conditions to those described in example 8. The product was purified in an identical manner to yield 56 mg of a white powder. MS(FAB+) m/e 878 (M+Na)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 0.95-1.52 (m, 6H), 1.31 (br s,9H), 2.22-2.32 (m, 4H), 2.60-3.20 (m,HH), 4.08-5.22 (m,4H), 6.62 (d, J=9Hz, 2H), 6.72-7.88 (m, 15H). Anal calcd for C_{4 5}H₅₇N₇O₁₀ CH₃COOH: C 60.44, H 6.80, N 10.50; found:

C 60.18, H 6.47 N 10.86.

Example 113

BOC-D-Trp-Lys (epsilon-N-(4-hydroxycinnamoyl))-Asp-N- alpha-Me-Phe-NH₂

The tetrapeptide of example 111 (60 mg), active ester of example 15 (28 mg) and N-methylmorpholine (9 mg) were allowed to react under similar conditions to those

described in example 8. The product was isolated in an identical manner to yield 55 mg of a white solid.

MS(FAB+) m/e 876 (M+Na)⁺. ¹H NMR(DMSO-d6, 300MHz) δ 1.10-1.44 (m, 6H), 1.31 (br s, 9H), 2.22-2.35 (m,2H), 2.68-3.23 (m, 9H), 4.08-5.22 (m,4H), 6.40 (d, J=16Hz, 1H), 6.72-6.84 (m, 3H), 6.92-7.62 (m, 12H), 7.81-8.16 (m, 3H). Anal calcd for C₄₅H₅₅N₇O₁₀ CH₃COOH: C 60.86, H 6.57, N 10.57; found: C 60.54, H 6.32, N 10.94.

Example 114

BOC-alpha-Nal-NH₂

The amide was prepared from BOC-alpha-Nal employing the mixed anhydride conditions described in example 97.

Example 115

alpha-Nal-NH₂ Hydrochloride

The amino acid was obtained from the deprotection of example 114 using similar conditions to those described in example 2.

Example 116

BOC-Asp (beta-Bn)-alpha-Nal-NH₂

The dipeptide was prepared via coupling of BOC-Asp(beta-Bn) with the amino acid of example 115 employing the conditions described in example 1.

Example 117

Asp (beta-Bn)-alpha-Nal-NH₂ Hydrochloride The deprotection of example 116 was conducted under similar conditions to those described in example 2.

Example 118

BOC-Lys (epsilon-N-Cbz)-Asp(beta-Bn)-alpha-Nal-NH₂

The tripeptide was prepared from coupling of BOC-Lys (epsilon-N-Cbz) with example 117 under conditions described for example 3.

Example 119

Lys ( epsilon-N-Cbz ) -Asp (beta -Bn ) -alpha-Nal -NH₂

Hydrochloride

The compound was prepared from example 118 in an identical manner to example 4.

Example 120

BOC-Trp-Lys (epsilon-N-Cbz)-Asp(beta-Bn)-alpha-Nal-NH₂ The tetrapeptide was prepared by coupling BOC-Trp with tripeptide of example 119 using the conditions described in example 5.

Example 121

BOC-Trp-Lys-Asp-alpha-Nal-NH₂

The compound was prepared from example 120 using the conditions described for example 6.

Example 122

BOC-Trp-Lys (epsilon-N-(3-(4-hydroxyphenyl) propionyl))-Asp-alpha-Nal-NH₂

The product was prepared from example 121 in a similar manner to that described for example 9. MS(FAB+) m/e 892 (M+H)⁺. ¹H NMR(DMSO-d6, 300MHz) δ 1.05-1.79

(m, 6H), 1.29 (br s,9H), 2.28 (t, J=8Hz, 2H), 2.40-2.72

(m,4H), 2.85-3.64 (m, 6H), 4.15-4.26 (m,2H), 4.42-4.55 (m,2H), 6.63 (d, J=8Hz, 1H) , 6.83-8.35 (m,16H), 9.14 (s,1H). Anal calcd for C₄₈H₅₇N₇O₁₀ 0.5H₂O: C 63.98, H 6.49, N 10.88; found: C 63.86, H 6.44, N 10.75.

Example 123

2-Hydroxycinnamic acid N-hydroxysuccinimide ester

A solution of 2-hydroxycinnamic acid (1.00 g), N-hydroxysuccinimide (0.72 g) and EDCI (1.20 g) in methylene chloride (15 mL) was stirred at ambient temperature for 18 h. The product was isolated in an identical manner as described in example 7. MS(CI/NH₃) m/e 262 (M+H)⁺. ¹H NMR (CDCl₃, 300MHz) δ 2.90 (br s,4H), 6.72 (d, J=15Hz, 1H), 6.81 (dd, J=7Hz,1H), 6.93 (t, 1H), 7.28 (t,1H), 7.38

(dd, J=7Hz,1H), 8.03 (d, 1H).

Example 124

BOC-Trp-Lys (epsilon-N-(2-hydroxycinnamoyl))-Asp-Phe-NH₂ The tetrapeptide of example 6 (100 mg), active ester of example 123 (38 mg) and N-methylmorpholine (28 mg) were allowed to react under identical conditions to those described in example 8. The product was isolated in a similar manner to yield 49 mg of a white solid. MS (FAB+) m/e 840 (M+H)⁺. ¹H NMR(DMSO-d6,300MHz) δ 1.04-1.66

(m,6H), 1.30 (br s,9H), 2.38-2.51 (m,2H), 2.58-2.69

(m,2H), 2.79-3.17 (m,4H), 4.16-4.42 (m, 3H), 4.50 (br q, 1H), 6.65 (d, J=16Hz,1H), 6.77-7.44 (m,15H), 7.58 (br s,1H), 7.62 (d,1H), 7.95 (d, 1H) , 8.06 (br t,1H), 8.28 (d, 1H), 10.83 (br s,1H). Anal calcd for C₄₄H₅₃N₇O₁₀ 1.75H₂O: C 60.64, H 6.54, N 11.25; found: C 60.48, H 6.36, N 10.97.

Example 125

3-Chlorocinnamic acid N-hydroxysuccinimide ester

A solution of 3-chlorocinnamic acid (1.00 g), N-hydroxysuccinimide (0.65 g) and EDCI (1.15 g) in methylene chloride (10 mL) was stirred at ambient temperature overnight. The product was isolated as described in example 7. MS(CI/NH₃) m/e 280 (M+H)⁺. ¹H

NMR (CDCl₃, 300MHz) δ 2.88 (br s,4H), 6.60 (d, J=16Hz, 1H), 7.32-7.49 (m,3H), 7.55 (t,1H), 7.85 (d, 1H).

Example 126

BOC-Trp-Lys (epsilon-N-(3-chlorocinnamoyl))-Asp-Phe-NH₂ The tetrapeptide of example 6 (100 mg), active ester of example 125 (41 mg) and N-methylmorpholine (28 mg) were allowed to react under similar conditions to those

described in example 8. The product was isolated in an identical manner to yield 64 mg of a white powder.

MS(FAB+) m/e 758 (M+H)⁺. ¹H NMR(DMSO-d6, 300MHz) δ 1.08-1.65 (m, 6H) , 1.29 (br s,9H), 2.35-2.65 (m,2H), 2.76-3.29 (m, 6H) , 4.16-4.40 (m, 3H) , 4.48 (br q, 1H), 6.69

(d, J=16Hz,1H), 6.83 (br d, J=8Hz, 1H), 6.91-7.62 (m, 14H), 7.92-8.05 (m,2H), 8.16 (br t,1H), 8.26 (br d, 1H), 10.85 (br s,1H). Anal calcd for C₄₄H₅₂N₇O₉Cl 1.25H₂O: C 59.99, H 6.24, N 11.13; found: C 59.78, H 6.09, N 11.08.

Example 127

BOC-Trp-Lys (epsilon-N-((6-OSO₃H)-2-naphthoyl))- Asp-Phe-NH₂

A solution of the tetrapeptide of example 73 (120 mg) and pyridinium acetyl sulfate (310 mg) in DMF and pyridine was allowed to react under the conditions described in example 10. The product was isolated via reverse phase

HPLC under identical conditions to yield a white powder after lyopholization. MS (FAB-) m/e 942 (M-H)-. ¹H

NMR (DMSO-d6, 500MHz) δ 1.05-1.69 (m, 6H), 1.29 (br s,9H), 2.42-2.52 (m, 1H), 2.62-2.68 (m, 1H), 2.92-3.16 (m, 6H), 4.20-4.32 (m,2H), 4.35-4.41 (m, 1H) , 4.51 (q, 1H), 6.81 (d, J=5Hz,1H), 6.95 (t, J=4Hz, 1H), 7.02-7.36 (m, 10H), 7.40 (dd, J=5Hz, 1H), 7.58 (d,1H), 7.70 (d, 1H), 7.81-7.95 (m, 3H), 8.25 (d, 1H), 8.38 (s, 1H), 8.53 (t,1H), 10.76 (s,1H) . Anal calcd for C₄₆H₅₂N₇O₁₃S 3H₂O NH₃ : C 54.48, H 6.06, N

11.05; found: C 54.13, H 5.66, N 10.86.

Example 128

2,4-Dimethoxycinnamic acid N-hydroxysuccinimide ester A solution of 2,4-dimethoxycinnamic acid (1.00 g), N-hydroxysuccinimide (0.56 g) and EDCI (0.95 g) in methylene chloride was allowed to stir at ambient temperature for 18 h. The product was isolated as described in example 7 to yield 1.04 g of a white solid. MS(CI/NH₃) m/e 306 (M+H)⁺. ¹H NMR(CDCl₃, 300MHz) δ 2.87 (br s,4H), 3.85 (s,3H), 3.88 (s,3H), 6.46 (d, J=2Hz,1H), 6.52 (dd, J=9Hz, 1H), 6.62

(d, J=16Hz,1H), 7.46 (d, 1H), 8.09 (d, 1H).

Example 129

BOC-Trp-Lys (epsilon-N-(2,4-dimethoxycinnamoyl))- Asp-Phe-NH₂

The tetrapeptide of example 6 (100 mg), active ester of example 128 (44 mg) and N-methylmorpholine (29 mg) were allowed to react under the conditions described in example 8. The product was isolated in a similar manner to yield 77 mg of a white solid. MS(FAB+) m/e 884 (M+H)⁺. ¹H NMR(DMSO-d6, 500MHz) δ 1.06-1.61 (m, 6H) , 1.28 (br s,9H), 2.36-2.43 (m,1H), 2.52-2.62 (m, 1H), 2.78-2.92 (m, 2H), 3.00-3.15 (m,4H), 3.77 (s,3H), 3.80 (s,3H), 4.14-4.28 (m,2H), 4.30-4.37 (m, 1H) , 4.46 (q, 1H), 6.47-6.57 (m, 3H), 6.78 (d,1H), 6.92 (t, 1H), 7.02 (t,1H), 7.05-7.40 (m,7H), 7.51-7.58 (m, 2H), 7.88-7.98 (m, 3H) , 8.21 (d, 1H), 10.80 (br s,1H). Anal calcd for C₄₆H₅₇N₇O₁₁ 0.5H₂O : C 61.87, H 6.55, N 10.98; found: C 61.47, H 6.43, N 10.84.

Example 130

4-(2-Naphthyl)-3-butenoic acid

N-hydroxysuccinimide ester

A solution of 4-(2-naphthyl)-3-butenoic acid (300 mg), N-hydroxysuccinimide (170 mg) and EDCI (310 mg) in methylene chloride was stirred at ambient temperature for 18 h. The product was isolated as described in example 7 to yield 190 mg of a white solid. MS(CI/NH-J m/e 309 M⁺. H NMR (CDCl₃, 300MHz) δ 2.84 (br s,4H), 3.62

(dd, J=7Hz, J=2Hz,2H), 6.38 (dt,1H), 6.79 (d, J=15Hz, 1H), 7.40-7.50 (m, 2H), 7.59 (dd, J=10Hz, J=2Hz, 1H), 7.70-7.85 (m, 4H).

Example 131

BOC-Trp-Lys (epsilon-N-(4-(2-naphtyl)-3-butenoyl))- Asp-Phe-NH₂

The tetrapeptide of example 6 (100 mg), active ester of example 130 (45 mg) and N-methylmorpholine (30 mg) were allowed to react under similar conditions to those

described in example 8. The product was purified in an identical manner to yield 77 mg of a white solid.

MS(FAB+) m/e 888 (M+H)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 1.06-1.66 (m,6H), 1.30 (br s,9H), 2.38-3.35 (m, 12H), 4.17-4.41 (m,3H), 4.50 (br q,1H), 6.41-6.65 (m, 2H), 6.85 (br d, 1H), 6.92-7.98 (m,18H), 8.28 (br d,1H), 10.32 (br s,1H). Anal calcd for C₄₉H₅₇N₇O₉ H₂O: C 64.95, H 6.56, N 10.82;

found: C 64.51, H 6.44, N 10.63.

Example 132

BOC-Lys-Asp-Phe-NH₂

A suspension of tripeptide of example 3 (150 mg) and 10% Pd-C (75 mg) in acetic acid was stirred under a balloon of hydrogen gas at ambient temperature. After 2 h, the reaction was filtered through Celite, the acetic acid was removed in vacuo and the resulting residue was dissolved in water then lyophilized to yield 107 mg of a white flocculent powder. MS (FAB+) m/e 508 (M+H)⁺.

Example 133

BOC-Lys (epsilon-N-(3-(4-hydroxyphenyl)propionyl))- Asp-Phe-NH₂

A solution of the tripeptide of example 132 (92 mg), N-methylmorpholine (23 mg) and 3-(4-hydroxyphenyl)propionic acid N-hydroxysuccinimide ester (55 mg) in N,N-dimethylformamide (5 mL) was stirred at ambient temperature for 18 h. The product was isolated under similar conditions to those described in example 8 to yield 50 mg of a white solid. MS (FAB+) m/e 656 (M+H)⁺. ¹H NMR (DMSO-d6, 500MHz) δ 1.10-1.57 (m, 6H), 1.37 (br s, 9H), 2.32 (t, J=7Hz,2H), 2.46-2.54 (m, 1H), 2.60-2.67 (m, 1H), 2.71 (t,H), 2.80-2.87 (m, 1H), 2.94-3.09 (m,3H), 3.86

(m,1H), 4.36 (m, 1H), 4.47 (br t, 1H), 6.68 (d,J=9Hz, 2H), 7.01 (d,2H), 7.17-7.32 (m, 6H), 7.44 (m, 1H) , 7.83 (m, 1H), 8.56 (br s,1H). Examole 134

Lys (epsilon-N-(3-(4-hydroxyphenyl)propionyl))- Asp-Phe-NH₂ HCl

The tripeptide of example 133 was reacted with HCl(g) in acetic acid as described in example 2. MS(FAB+) m/e 556 (M+H)⁺. ¹H NMR (DMSO-d6, 500MHz)

d 1.15-1.25 (m,2H), 1.29-1.37 (m,2H), 1.58-1.66 (m,2H), 2.34 (t, J=8Hz,2H), 2.51-2.61 (m, 1H), 2.68-2.79 (m, 3H), 2.81-2.88 (m, 1H), 2.95-3'.08 (m, 4H), 3.69 (t, J=7Hz, 1H), 4.41 (m,1H), 4.60 (t,1H), 6.69 (d, J=8Hz, 2H), 7.02 (d,2H), 7.19-7.32 (m,6H), 7.89 (t,1H), 8.39 (t,1H), 8.75 (br d,1H).

Example 135

Ctp-Lys (epsilon-N-(3-(4-hydroxyphenyl)propionvl))- Asp-Phe-NH₂

To a solution of Ctp acid (24 mg; Ctp: Tryptophan-4-acetic acid lactam; M. Mascal, C. J. Moody, J. Chem. Soc., Chem. Commun., 587 (1987)) in tetrahydrofuran (5 mL) cooled to -15ºC were added N-methylmorpholine (10 mg) and

isobutylchloroformate (13 mg) . After stirring five minutes at -15 °C, a solution of tripeptide of example 134 (37 mg) and N-methylmorpholine (10 mg) in N,N-dimethylformamide (5 mL) was added to the reaction mixture. The solution was stirred for 2 h at -15°C then for 18 h at ambient

temperature. The solvents were removed in vacuo and the residue was purified as described in example 8. MS(FAB+) m/e 782 (M+H)⁺. ¹H NMR (DMSO-d6, 500MHz) δ 1.16-1.61 (m, 6H), 2.27 (t, J=8Hz,2H), 2.38-2.58 (m, 1H), 2.62-2.84 (m,3H), 2.93-3.57 (m, 8H), 4.16 (m,2H), 4.27 (m,2H), 6.64 (d, J=9Hz, 1H), 6.75 (d, J=8Hz,1H), 6.92-7.02 (m, 6H) , 7.08-7.23 (m, 6H), 7.32-7.47 (m, 1H), 7.62-7.86 (m, 2H), 8.19-8.38 (m, 2H), 10.92 (br S,1H).

Example 136

2-Naphthoxyacetyl-Lys (epsilon-N-Cbz)-ASP(beta-Bn)-Phe-NH₂ A solution of tripeptide of example 4 (300 mg), 2-naphthoxyacetic acid N-hydroxysuccinimide ester (161 mg) and N-methylmorpholine (50 mg) in N,N-dimethylformamide (10 mL) was stirred at ambient temperature for 18 h. The solvent was removed in vacuo and the residue was dissolved in 25% isopropanol in chloroform which was washed successively with 1M phosphoric acid solution, saturated sodium bicarbonate solution and brine. After drying over sodium sulfate, the solvent was removed in vacuo to yield a white solid.

MS(FAB+) m/e 816 (M+H)⁺. ¹H NMR(DMSO-d6,300MHz) δ 1.14-1.74 (m,6H), 2.51-2.69 (m, 2H), 2.81-3.19 (m, 6H), 4.31-4.48

(m,2H), 4.61-4.75 (m, 3H), 5.04 (s,2H), 5.11 (s,2H), 7.16-7.55 (m,20H), 7.79-7.95 (m, 4H), 8.22 (d,1H), 8.46 (d, 1H).

Example 137

2-Naphthoxyacetyl-Lys-Asp-Phe-NH₂

The tripeptide of example 136 was treated in a similar manner as described in example 6. MS(CI/NH₃) m/e 592

(M+H)⁺.

Example 138

2-Naphthoxyacetyl-Lys (epsilon-N-(4-hydroxycinnamoyl))- Asp-Phe-NH₂

The tripeptide of example 137 (40 mg), active ester of example 15 (23 mg) and N-methylmorpholine (8 mg) were allowed to react under the conditions described in example 8. The product was purified in an identical manner to yield 22 mg of a white solid. MS(FAB+) m/e 738 (M+H)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 1.12-1.71 (m, 6H), 2.26-2.52

(m,2H), 2.74-2.86 (m, 2H) , 2.99-3.12 (m, 4H) , 4.24-4.35 (m, 2H), 4.42 (br q, 1H), 4.68 (br s,2H), 6.42

(d, J=16Hz,1H), 6.77 (d, J=8Hz, 2H), 7.02-7.64 (m, 12H), 7.75- 7.87 (m,3H), 8.03 (br t,1H), 8.15 (br d, 1H), 8.22 (d, 1H), 8.38 (d,1H). Anal calcd for C₄₀H₄₃N₅O₉ 2HCO₂H: C 60.79, H 5.71, N 8.44; found: C 60.99, H 5.60, N 8.68.

Example 139

3-(3-Indolyl)propionic acid 2,4,5-trichlorophenol ester A solution of 3-(3-indolyl)propionic acid (9.5 g) and 2,4,5-trichlorophenol (10.4 g) in ethyl acetate (65 mL) was stirred at ice bath temperature for 4 h. The

resulting precipitate was filtered off and the solvent was evaporated in vacuo. The residue was crystallized from ethanol to yield light brown needles. m.p. 105-105.5 ºC.

Example 140

3-(3-Indolyl)propionyl-Lys(epsilon-N-Cbz)- Asp(beta-Bn)-Phe-NH₂

The tripeptide of example 4 (252 mg), active ester of example 139 (167 mg) and N-methylmorpholine (40 mg) were reacted under the conditions described in example 136. The product was isolated in an identical manner.

MS(CI/NH₃) m/e 803 (M+H)⁺.

Example 141

3-(3-Indolyl)propionyl-Lys-Asp-Phe-NH₂ The tripeptide of example 140 (100 mg) and 10% Pd-C (50 mg) were reacted and the product isolated as described in example 137 to yield 58 mg of a light brown powder. MS(CI/NH₃) m/e 579 (M+H)⁺. ¹H NMR(DMSO-d6, 300MHz) δ 1.15-1.68 (m,6H), 2.17-2.39 (m, 2H), 2.47-3.32 (m, 10H), 3.99-4.08 (m, 1H), 4.18-4.29 (m, 1H), 4.31-4.42 (m, 1H), 6.88-7.55 (m, 9H), 7.92 (d,1H), 8.13 (d,1H), 8.29 (d,1H), 10.80 (br s,1H).

Example 142

3-(3-Indolyl)propionyl-Lys (epsilon-N- (3-(4-hydroxyphenyl)propionyl))-Asp-Phe-NH₂ The tripeptide of example 141 (35 mg), 3-(4-hydroxyphenyl) propionic acid N-hydroxysuccinimide ester (19 mg) and N-methylmorpholine (7 mg) were allowed to react as described in example 7. The product was isolated in a similar manner to yield 35 mg of a white flocculent powder. MS (FAB+) m/e 727 (M+H)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 1.08-1.59 (m, 6H), 2.23-2.58 (m, 4H), 2.68 (t, J=8Hz, 2H), 2.79-3.22 (m,8H), 4.08-4.18 (m,1H), 4.23-4.39 (m,2H), 6.63 (dt,J=9Hz,1H), 6.89-7.28 (m, 8H) , 7.31 (d, 1H), 7.49-7.56 (m,2H), 7.80 (t,1H), 8.06-8.15 (m,2H), 8.26 (d, 1H). Anal calcd for C₃₉H₄₆N₆O₈ 1.5H₂O: C 62.14, H 6.55, N 11.15; found: C 61.73, H 6.22, N 10.89. Example 143

BOC-Trp-Lys (epsilon-N-3-(3-indolyl)propionyl)-Asp-Phe-NH.

The tetrapeptide of example 6 (75 mg) , active ester of example 139 (48 mg) and N-methylmorpholine (12 mg) were reacted under similar conditions to those described in example 8. The product was isolated in a similar manner to yield 46 mg of a white solid. MS(FAB+) m/e 887 (M+Na)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 1.08-1.63 (m, 6H) , 1.32 (br s, 9H), 2.38-2.68 (m,4H), 2.80-3.18 (m,10H), 4.18-4.30 (m, 2H), 4.32- 4.41 (m,1H), 4.49 (br q, 1H), 6.78 (br d, 1H), 6.90-7.61 (m,14H), 7.81 (br t,1H), 7.90-8.05 (m, 2H) , 8.20-8.30 (m,2H), 10.72 (br s,1H), 10.82 (br s,1H). Anal calcd for C₄₆H₅₆N₈O₉ H₂O: C 62.57, H 6.62, N 12.69; found: C 62.43, H 6.37, N 12.47.

Example 144

BOC-Trp-Lys (epsilon-N-(BOC-Tyrosyl))-Asp-Phe-NH₂

The tetrapeptide of example 6 (111 mg), BOC-Tyr N-hydroxysuccinimide ester (73 mg) and N-methylmorpholine (18 mg) were allowed to react under the conditions described in example 8. The product was purified in a similar manner to yield 96 mg of a white powder. MS(FAB+) m/e 955 (M+H)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 1.10-1.65 (m, 6H), 1.32 (br s,18H), 2.44-3.36 (m,12H), 4.00-4.12 (m, 1H), 4.25 (br s,2H), 4.35-4.44 (m,1H), 4.52 (br q, 1H), 6.60-6.82 (m,3H), 6.92-7.35 (m,12H), 7.58 (d,1H), 7.74-7.96 (m,3H), 8.21-8.29 (m,2H), 9.08 (br s,1H), 10.72 (br s,1H). Anal calcd for C₄₉H₆₄N₈O₁₂ 0.5H₂O: C 60.92, H 6.78, N 11.60; found: C 60.99 H 6.78, N 11.62. Example 145

BOC-Trp-Lys (epsilon-N-(BOC-O-sulfatyl-Tyrosyl))- Asp-Phe-NH₂

The tetrapeptide of example 144 (41 mg) and

pyridinium acetyl sulfate (94 mg) were reacted as

described in example 10. The product was isolated by preparative reverse phase HPLC as described to yield 26 mg of a white solid after lyophilization. MS (FAB+) m/e 1037 (M+H)⁺. ¹H NMR(DMSO-d6, 300MHz) δ 1.11-1.62 (m, 6H), 1.32 (br s,18H), 2.38-3.25 (m,12H), 4.04-4.15 (m, 1H), 4.18-4.29 (m, 2H), 4.32-4.42 (m, 1H), 4.45-4.55 (m, 1H) , 6.71-6.86 (m, 2H), 6.91-7.35 (m, 12H), 7.58 (d,1H), 7.80-7.95 (m,2H), 8.18-8.32 (m,2H), 10.75 (br s,1H). Anal calcd for

C₄₉H₆₄N₈O₁₅S 2·5H₂O NH₃ : C 53.54, H 6.60, N 11.47; found: C 53.39, H 6.20, N 11.24.

Example 146

BOC-Trp-Lys (epsilon-N-(BOC-Tryptophyl))-Asp-Phe-NH₂ The tetrapeptide of example 6 (70 mg), BOC-Trp N-hydroxysuccinimide ester (49 mg) and N-methylmorpholine (11 mg) were allowed to react under the conditions described in example 8. The product was isolated in an identical manner to yield 44 mg of a white flocculent solid. MS(FAB+) m/e 980 (M+H) ⁺ . ¹H NMR(DMSO-d6, 300MHz) δ 1.06-1.65 (m,6H), 1.32 (m, 9H), 2.39-3.19 (m, 12H), 4.12-4.29 (m,3H), 4.32-4.42 (m,1H), 4.46-4.55 (m,1H), 6.64 (br s,1H), 6.75 (br s,1H), 6.90-7.41 (m,15H), 7.58 (br d, 1H), 7.80-8.01 (m,3H), 8.18-8.30 (m,2H), 10.76 (br s,1H), 10.80 (br s,1H). Anal calcd for C₅₁H₆₅N₉O₁₁ 1.5H₂O: C 60.82, H 6.80, N 12.52; found: C 60.85, H 6.57, N 12.28. Example 147

L-3-Benzyloxycarbonyl-5-Oxo-4-Oxazolidinepropionic Acid To a suspension of CBZ-glutamic acid (4.22 g, 15.0 mmol) in toluene (110 mL) were added 37% formalin solution (4.5 mL) and para-toluenesulfonic acid monohydrate (0.21 g, 1.1 mmol). The mixture was subsequently heated to reflux and water removed via toluene azeotrope (1-1.5 h). The crude product was washed with water (3X), dried (MgSO) and concentrated in vacuo. Purification by flash

chromatography on silica gel (ethyl acetate-hexane-acetic acid) gave 3 . 6 g of a clear oil. H NMR(CDCl₃, 300MHz) δ 2.2 (cm,1H), 2.32 (cm, 1H), 2.52 (br m, 2H) , 4.41 (m, 1H), 5.19 (s,2H), 5.23 (d, 1H), 5.55 (br s, 1H), 7.35 (s,5H). Anal calcd for dicyclohexylammoniun salt C₂₆H₃₈N₂O₆: C 65.79, H 8.09, N 5.90; found: C 65.50, H 8.01, N 5.74.

Example 148

L-3-Benzyloxycarbonyl-5-Oxo-4- Qxasolidinepropionic Acid Chloride To a solution of the product from example 147 (2.32 g, 7.91 mmol) in anhydrous dichloromethane (50 mL) at room temperature was added thionyl chloride (2.0 mL, 26.9 mmol). The mixture was allowed to stir overnight

protected from atmospheric moisture. The volatiles were removed in vacuo, the resulting residue dissolved in benzene and concentrated in vacuo (2X) to yield a solid product (2.26 g) of high purity. ¹H NMR(CDCl₃, 300MHz) δ 2.25 (br m,2H), 3.08 (cm,2H), 4.37 (m, 1H) , 5.21 (m,3H),

5.54 (br s,1H), 7.3-7.5 (m,5H). Anal calcd for

C₁₄H₁₄NO₅Cl: C 53.93, H 4.54, N 4.49; found: C 54.24, H

4.59, N 4.53, Example 149

L-3-Benzyloxycarbonyl-5-Oxo-4-Oxazolidinepent-2-enoic

Acid tert-Butyl Ester

The product from example 148 (0.6 g, 1.92 mmol) was dissolved in anhydrous 1,2-dimethoxyethane (26 mL) under nitrogen and the solution cooled to -78 °C. To this was added dropwise a solution of lithium tri-tertbutoxy aluminum hydride (0.51 g, 2 mmol) in anhydrous 1,2-dimethoxyethane (18 mL). Upon completion of the addition, the mixture was stirred 15 min at -78ºC before warming to room temperature. After one half hour the reaction was quenched by the addition of 10% aqueous HCl (1 mL) and the mixture poured into acidified brine. The crude product was isolated by extraction with ethyl acetate (3X). The combined extracts were washed with 5% sodium bicarbonate solution (2X), water (IX) then dried (MgSO₄) and

concentrated in vacuo to yield a clear oil (0.334 g). In practice the aldehyde is sufficiently pure to be used directly. To a solution of the aldehyde prepared as above (0.169 g, 0.61 mmol) in anhydrous dichloromethane (12 mL) at room temperature was added tertbutoxycarbonylmethylenetriphenyl phosphorane (0.46 g, 1.22 mmol) prepared according to Kenner,G.W.,et.al.,

Tetrahedron, 32 (2), 275 (1976) in one portion. The resulting clear solution was allowed to stir overnight under a nitrogen atmosphere. The product was isolated by concentration in vacuo followed by flash chromatography on silica gel (ethyl acetate-hexane) to give a white solid (0.154 g). ¹H NMR(CDCl₃, 300MHz) δ l.48 (s,9H), 2.05 (br m, 2H), 2.26 (br m,2H), 4.35 (cm,1H), 5.2 (cm,3H), 5.53 (br s , 1H) , 5 . 97 (br d, 1H J=15Hz ) , 6 . 77 (br m, 1H) , 7 . 3-7 . 45 (m, 5H) .

Example 150

L-N-alpha-CBZ,alpha-Amino,trans-5.6-Dehydropimelic acid alpha Methyl, epsilon-tert-Butyl Diester The product from example 149 (0.15 g, 0.386 mmol) was dissolved in methanol (5 ml) and the solution chilled in an ice bath. To this was added sodium (11.3 mg, 0.50 mmol). Following consumption of the sodium the mixture was warmed to room temperature. The product was isolated by acidification with cold, dilute aqueous HCl followed by chloroform extraction. The combined extracts were dried (MgSO₄) and concentrated in vacuo to give a quantitative yield of the diester as an oil sufficiently pure for use in subsequent chemistry. ¹H NMR(CDCl₃, 300MHz) δ 1.47 (s,9H), 1.8 (cm,1H), 2.07 (cm,1H), 2.23 (cm,2H), 3.75 (s,3H), 4.42 (cm,1H), 5.12 (s,2H), 5.32 (d, 1H) , 5.75 (d, 1H J=15Hz), 6.8 (dt,1H), 7.3-7.45 (m, 5H).

Example 151

L-N-alpha-CBZ, alpha-Amino. trans-5,6-Dehvdropimelic

acid alpha-Methyl Ester

The product from example 150 (1.96 g, 5.0 mmol) was dissolved in dichloromethane (50 ml) and treated at room temperature with a total of 6.6 equivalents of

trifluoroacetic acid (2.4 mL). Upon completion of the deprotection the mixture was washed with water (IX) then dried (MgSO₄) and concentrated in vacuo to give a solid product in quantitative yield sufficiently pure for use in subsequent chemistry. Example 152

L-N-alpha-CBZ,alpha-Amino,trans-5,6-Dehydropimelic acid alpha-Methyl Ester, epsilon-Tyramide The product from example 151 (0.10 g, 0.31 mmol) was dissolved in anhydrous THF (10 mL) under nitrogen and treated with N-methylmorpholine (0.037 ml, 0.34 mmol) followed by cooling to -10/-15 ºC. To the cold solution was added isobutylchloroformate (0.040 ml, 0.31 mmol). After ten minutes a previously prepared solution of tyramine (0.045 g, 0.31 mmol) in THF (4 mL) /DMF (1 mL) was added dropwise over approximately five minutes. Upon completion of the addition, the mixture was warmed to 0/+5

C. The product was isolated by partitioning between dilute aqueous HCl and ethyl acetate. The organic phase was washed (IX) with water then dried (MgSO₄) and

concentrated in vacuo. The crude product was purified by flash chromatography on silica gel (ethyl acetate-hexane-acetic acid) to yield an oil (0.063g). MS(CI/NH₃) m/e 441 (M+H)⁺, m/e 458 (M+NH₄)⁺. ¹H NMR (DMSO-d6,300MHz) δ 1.75 (cm,2H), 2.18 (cm,2H), 2.59 (cm,2H), 3.63 (s,3H), 4.03 (cm,1H), 5.04 (s,2H), 5.87 (d, 1H J=15Hz), 6.55 (dt,1H), 6.67 (d,2H), 6.98 (d,2H), 7.25-7.45 (cm,5H), 7.8 (d,1H), 7.96 (t,1H).

Example 153

L-N-alpha-CBZ,alpha-Amino,trans-5,6-Dehydropimelic acid,epsilon-Tyramide

The product from example 152 was dissolved in methanol (5 mL) and treated at room temperature under a nitrogen atmosphere with 0.1N KOH solution (4.4 mL). The crude product was isolated by concentration in vacuo followed by partitioning between dilute aqueous HCl and ethyl acetate. The organic phase was washed (IX) with water, dried (MgSO₄) and concentrated in vacuo to give in quantitative yield an oil which was sufficiently pure for use in subsequent chemistry.

Example 154

L-N-alpha-CBZ,alpha-Amino,trans-5,6-Dehydropimelic

acid (epsilon-Tyramide)-Asp(beta-Bn)-Phe-NH₂

The product from example 153 (0.129 mmol) was combined with the TFA salt of the product from example 2 (0.063 g, 0.129 mmol), 1-hydroxybenzotriazole (0.020 g, 0.129 mmol) and EDCI (0.027 g, 0.142 mmol). Anhydrous DMF (1 ml) was added and the flask capped with a calcium chloride filled drying tube. The solution was chilled in an ice bath and to the cold solution was added diisopropylethylamme (0.025 mL, 0.142 mmol) . The reaction mixture was allowed to warm to room temperature. The crude product was isolated by partitioning between ethyl acetate and dilute aqueous HCl. The organic phase was washed (IX) with water and then concentrated to dryness in vacuo. The crude product was dissolved in glacial acetic acid, frozen and lyophilized to yield 0.091 g of a white solid sufficiently pure for use in subsequent chemistry. ¹H NMR(DMSO-d6, 300MHz) δ 1.6 (cm,2H), 2.1 (cm, 2H), 2.6 (cm,2H), 3.97 (cm, 1H), 4.37 (cm, 1H) , 4.62 (cm, 1H), 4.9-5.1 (m, 4H) , 5.86 (d, 1H J=15Hz), 6.5-6.64

(dt,1H), 6.67 (d,2H), 6.98 (d,2H), 7.52 (d, 1H), 7.90

(d, 1H), 8.34 (d,1H).

Example 155

alpha-Aminopimelic acid-(epsilon-Tyramide)-Asp- Phe-NH₂ Hydrochloride

A mixture of the product from example 154 (0.086 g, 0.111 mmol) and 10% Pd/C (0.086 g) in glacial acetic acid (3 mL) was hydrogenated "overnight at room temperature and four atmospheres hydrogen pressure. The product was isolated by vacuum filtration through celite. The filtrate was frozen and lyophilized to yield a glass which was treated with 1.5 N HCl in glacial acetic acid to give, after lyophilization, a solid hydrochloride in quantitative yield.

Example 156

BOC-Trp-alpha-Aminopimelic acid-(epsilon-Tyramide)-

Asp-Phe-NH₂

The product from example 155 (0.11 mmol) was combined with BOC-Trp hydroxysuccinimide ester in anhydrous DMF (2 mL) . To this solution was added diisopropylethylamme (0.043 ml, 0.24 mmol) and the mixture allowed to stir overnight at room temperature. The crude reaction mixture was chromatographed (preparative HPLC) on a Vydac C₁₈ column (acetonitrile-0.05 M ammonium acetate @ pH 4.5) to afford after lyophilization, the desired product (0.013 g) as a white solid. An additional amount (0.005 g) was recovered from a recycle of mixed fractions. MS(FAB+) m/e 842 (M+H)⁺, m/e 864 (M+Na)⁺. ¹H NMR(DMSO-d6, 300MHz) δ 1.0-1.7 (cm,15H), 2.0 (t,2H), 4.0-4.7 (cm,4H), 6.66 (d,2H), 6.83 (d,2H), 6.9-7.5 (m, 11H), 7.59 (d, 1H) , 7.81 (t, 1H), 7.93 (d,2H), 8.26 (d, 1H) , 10.82 (s,1H). Anal calcd for C₄₄H₅₅N₇O₁₀ 3.5 H₂O: C 58.38, H 6.92, N 10.83; found : C 58.21, H 6.10, N 10.69.

Example 157

BOC-Trp-Lys (epsilon-N-(2-methylphenylaminocarbonyl))-

Asp-Phe-NH₂

A solution of tetrapeptide of example 6 (50 mg) , 2-methylphenyl isocyanate (15 mg) and N-methylmorpholine (18 mg) in DMF (5 mL) was stirred at ambient temperature for 18 h. The product was isolated as described in example 8 to yield 36 mg of a white solid. MS(FAB+) m/e 827 (M+H)⁺ . ¹H NMR (DMSO-d6, 300MHz) δ 1.05-1.69 (m, 6H) , 1.30 (br s,9H), 2.16 (s,3H), 2.35-3.25 (m, 10H), 4.16-4.40 (m, 3H), 4.48 (q,1H), 6.78-7.42 (m, 16H) , 7.59 (br d, J=7Hz, 1H), 7.75-8.05 (m,4H), 8.26 (d, 1H), 8.58 (m, 1H) , 10.82 (br s,1H). Anal calcd for C₄₃H₅₄N₈O₉ CH₃CO₂H: C 60.94, H 6.59, N 12.63; found: C 60.92, H 6.58, N 12.54.

Example 158

3-(Cyclohexyl)propionic acid N-hydroxysuccinimide ester A solution of 3-(cyclohexyl)propionic acid (0.72 g), N-hydroxysuccinimide (0.55 g) and EDCI (0.90 g) in

methylene chloride (20 mL) was stirred at ambient

temperature for 18 h. The active ester was isolated as described in example 7 to yield 0.60 g of a white solid. MS(CI/NH₃) m/e 271 (M+NH₄)⁺. ¹H NMR (CDCl₃, 300MHz) δ 0.82-1.01 (m,2H), 1.08-1.40 (m, 4H) , 1.59-1.78 (m, 7H), 2.61

(t, J=8Hz,2H), 2.84 (br s,4H).

Examole 159

BOC-Trp-Lys (epsilon-N- (3- (cyclohexyl) propionyl) ) - Asp-Phe-NH₂

A solution of tetrapeptide of example 6 (100 mg), active ester of example 158 (37 mg) and N-methylmorpholine

(28 mg) in DMF was allowed to react as described in example 8. The product was isolated in an identical fashion to yield 68 mg of a white solid. MS (FAB+) m/e 854 (M+Na)⁺. ¹H NMR(DMSO-d6, 300MHz) δ 0.71-0.90 (m,2H), 1.02-1.40

(m,10H), 1.50-1.69 (m,7H), 2.04 (t, J=8Hz, 2H) , 2.38-3.18

(m,10H), 4.12-4.29 (m, 2H) , 4.32-4.40 (m, 1H) , 4.50 (q,1H), 6.84 (br d, J=8Hz,1H), 6.92-7.35 (m, 10H) , 7.59 (d, 1H), 7.71

(br t,1H), 7.90 (m,2H), 8.26 (d, 1H) , 10.80 (br s,1H). Anal calcd for C₄₄H₆₁N₇O₉ H₂O: C 62.17, H 7.47, N 11.54; found: C 62.44, H 7.26, N 11.46.

Example 160

BOC-Trp-Lys (epsilon-N-(8-hydroxyquinolyl-2-carbonyl))- Asp-Phe-NH₂

A solution of 8-hydroxyquinolyl-2-carboxylic acid (50 mg), N-hydroxysuccinimide (31 mg) and EDCI (60 mg) in DMF (4 mL) was stirred at ambient temperature for 18 h. The active ester was not isolated but immediately reacted with tetrapeptide of example 6 (88 mg) and N-methylmorpholine (58 mg) under the conditions described in example 8. The product was purified in a similar manner to yield 62 mg of a white solid. MS(FAB+) m/e 865 (M+H)⁺. ¹H NMR(DMSO-d6, 500MHz) δ 1.02-1.68 (m, 6H), 1.25 (br s, 9H), 2.45

(dd,J=6Hz,J=16Hz,1H), 2.63 (dd,1H), 2.78-2.91 (m, 4H), 2.95- 3.15 (m,4H), 4.19 (br s,1H), 4.25 (br s,1H), 4.33 (m, 1H), 4.48 (q,1H), 6.78 (d,2H), 6.88-7.30 (m, 15H) , 7.42 (d, 1H), 7 . 52 (t , 2H) , 7 . 78 (d, 1H) , 7 . 88 ( d, 1H) , 8 . 22 (d, 1H) , 9 . 57

(br t,1H). Anal calcd for C₄₅H₅₂N₈O₁₀ H₂O: C 61.22, H 6.16, N 12.69; found: C 61.44, H 6.03, N 12.51.

Example 161

5-Methoxyindolyl-2-carboxylic acid

N-hydroxysuccinimide ester

A solution of 5-methoxyindolyl-2-carboxylic acid (1.00 g), N-hydroxysuccinimide (0.61 g) and EDCI (1.10 g) in methylene chloride was allowed to stir at ambient

temperature for 18 h. The active ester was isolated as described in example 7 to yield 0.55 g of a white solid. MS(CI/NH₃) m/e 289 (M+H)+. 1H NMR(CDCl₃, 300MHz) δ 2.91 (s,4H), 3.79 (s,3H), 7.04 (dd, J=3Hz, J=10Hz, 1H), 7.18

(d,1H), 7.39-7.45 (m,2H), 12.31 (br s,1H).

Example 162

BOC-Trp-Lys (epsilon-N-(5-methoxyindolyl-2-carbonyl))- Asp-Phe-NH₂

A solution of tetrapeptide of example 6 (100 mg), active ester of example 161 (45 mg) and N-methylmorpholine

(29 mg) was allowed to react as described in example 8.

The final product was isolated in a similar manner to yield 80 mg of a white solid. MS(FAB+) m/e 925 (M+H)⁺. ¹H

NMR (DMSO-d6, 500MHz) δ 1.08-1.68 (m, 6H), 1.29 (br s,9H), 2.42 (dd, J=7Hz, J=17Hz,1H), 2.58 (dd, 1H), 2.76-2.95 (m, 3H) , 3.02-3.30 (m,4H), 3.72 (s,3H), 4.25 (br s,2H), 4.36 (m, 1H) , 4.48 (q,1H), 6.75-6.82 (m,2H), 6.92-7.45 (m, 1H), 7.58

(d,1H), 7.95 (d,2H), 8.08 (d, 1H), 8.50 (br s,1H), 10.82 (br s,1H). Anal calcd for C₄₅H₅₄N₈O₁₀ 0.5H₂O 0.75CH₃CO₂H: C 60.57, H 6.35, N 12.12; found: C 60.69, H 6.59, N 12.01. Example 163

BOC-Trp-Lys (epsilon-N-BOC-D-Trptophyl)-Asp-Phe-NH₂

A solution of BOC-D-Trp (75 mg) and EDCI (25 mg) in methylene chloride and N,N-dimethylformamide was stirred at ambient temperature for 4 h. The tetrapeptide of example 6 (75 mg) and N-methylmorpholine (12 mg) were added to the above solution and allowed to react for 18 h. The product was isolated as described in example 8 to yield 60 mg of a white solid. MS (FAB+) m/e 1002 (M+Na)⁺. ¹H NMR(DMSO-d6, 300MHz) δ 1.06-1.68 (m, 6H) , 1.32 (br s,18H), 2.35-3.25 (m, 10H), 4.11-4.30 (m,3H), 4.32-4.42 (m, 1H) , 4.50 (br q, 1H), 6.68 (br d, 1H), 6.80 (br d, 1H), 6.91-7.38 (m, 14H), 7.58 (br d,1H), 7.82-7.90 (m, 3H), 8.21-8.31 (m, 2H), 10.78 (br s,1H), 10.80 (br s,1H). Anal calcd for C₅₁H₆₅N₉O₁₁ H₂O: C 61.37, H 6.77, N 12.63; found: C 61.28, H 6.48, N 12.59.

Example 164

BOC-Trp-Lys (epsilon-N-BOC-D-Tyrosyl)-Asp-Phe-NH₂

A solution of BOC-D-Tyr (70 mg) and EDCI (26 mg) in methylene chloride and DMF was stirred at ambient

temperature for 6 h. A solution of tetrapeptide of example 6 (58 mg) and N-methylmorpholine (9 mg) in DMF was added to the above reaction and allowed to stand for 18 h. The product was purified as described in example 8 to yield 43 mg of a white solid. MS(FAB+) m/e 957 (M+H)⁺. ¹H

NMR(DMSO-d6, 300MHz) δ 1.06-1.68 (m, 6H), 1.32 (br s,18H), 2.33-3.18 (m,10H), 4.04 (br s,1H), 4.24 (br s,2H), 4.32- 4.42 (m,1H), 4.49 (q, 1H), 6.62 (br d,2H), 6.66-6.80 (m, 2H), 6.91-7.42 (m,15H), 7.58 br d, 1H), 7.81 (br s,1H), 7.92-7.98 (m, 2H), 8.19-8.30 (m, 1H) , 10.80 (br s, 1H). Anal calcd for

C₄₉H₆₄N₈O₁₂ H₂O: C 60,35, H 6.82, N 11.49; found: C 60.28, H 6.71, N 11.36.

Example 165

5-(Benzyloxy)indole-2-carboxylic acid N-hydroxysuccinimide ester

A solution of 5- (benzyloxy) indole-2-carboxylic acid (1.00 g), N-hydroxysuccinimide (0.45 g) and EDCI (0.72 g) in methylene chloride was stirred at ambient temperature for 18 h. The resulting solid was collected, washed and dried to yield 1.26 g of a white solid. MS(CI/NH₃) m/e 365 (M+H)⁺. ¹H NMR (DMSO-d₆, 300MHz) δ 2.92 (br s,4H), 5.12 (s,2H), 7.12 (dd, J=2Hz, J=12Hz,1H), 7.28-7.52 (m, 8H), 12.32 (br S,1H).

Example 166

BOC-Trp-Lys (epsilon-N-(5-(benzyloxy)indole-2-carbonyl)- Asp-Phe-NH₂

The tetrapeptide in example 6 (100 mg), active ester in example 165 (55 mg) and N-methylmorpholine (32 mg) were allowed to react in a similar manner to that described in example 8. The final product was isolated under identical conditions to yield 85 mg of a white solid. MS (FAB+) m/e 943 (M+H)⁺. ¹H NMR (DMSO-d₆, 500MHz) δ 1.05-1.62 (m, 6H), 1.23 (br s,9H), 2.38 (dd, J=6Hz, J=16Hz, 1H), 2.52 (dd, 1H), 2.71-3.12 (m,6H), 4.20 (br s,2H), 4.32 (m, 1H) , 4.42 (br q,1H), 5.03 (s,2H), 6.71-7.57 (m, 10H), 7.92 (br d, 2H), 8.13 (d,1H), 8.49 (br s,1H). Anal calcd for C₅₁H₅₈N₈O₁₀ C₂H₄O₂ 0.5H₂O: C 62.90, H 6.27, N 11.07; found: C 62.79, H 6.16, N 11.08. Example 167

5-Chloroindole-2-carboxylic acid N-hydroxy- succinimide ester

A solution of 2-chloroindole-2-carboxylic acid (0.50 g), N-hydroxysuccinimide (0.30 g) and N-methylmorpholine (32 mg) in methylene chloride was stirred at ambient temperature for 18 h. The solvent was removed in vacuo and the residue was dissolved in ethyl acetate and washed with water. After drying over MgSO₄, the solvent was evaporated in vacuo and the resulting solid was dissolved in ethyl acetate and precipitated with the addition of hexane to yield 0.47 g of a white solid. MS(CI/NH₃) m/e 310

(M+NH₄)⁺. ¹H NMR(CDCl₃, 300MHz) δ 2.92 (br s,4H), 7.32-7.42 (m,3H), 7.69 (m, 1H), 9.12 (br s,1H).

Example 168

BOC-Trp-Lys (epsilon-N-(5-chloroindole-2-carbonyl))- Asp-Phe-NH₂

A solution of tetrapeptide in example 6 (100 mg), active ester in example 167 (44 mg) and N-methylmorpholine

(32 mg) in DMF was allowed to react under similar

conditions to those described in example 8. The product was isolated in an identical manner to yield 79 mg of a white solid. MS (FAB+) m/e 893 (M+Na)⁺. ¹H NMR (DMSO- d₆,500MHz) δ 1.08-1.68 (m, 6H) , 1.31 (br s,9H), 2.41-2.52

(m,1H), 2.58-2.68 (m, 1H) , 2.82-3.96 (m, 2H), 3.02-3.15

(m,2H), 4.25 (br s,2H), 4.36 (br q, 1H), 4.50 (br q, 1H), 6.79 (d, J=7Hz,1H), 6.95 (t,1H), 7.01-7.39 (m, 14H), 7.42

(d, J=9Hz,1H), 7.63 (s,1H), 7.84-7.97 (m,2H), 8.20 (br d,1H), 8.62 (br s,1H), 10.78 (s, 1H). Anal calcd for C₄₄H₅₁ClN₈O₉ 2H₂O: C 58.24, H 6-11 N 12-35; found:

58.30, H 5.91, N 12.20

Example 169

5-Hvdroxyindole-2-carboxylic acid N-hvdroxy- succinimitie ester

A solution of 5-hydroxyindole-2-carboxylic acid (0.50 g), N-hydroxysuccinimide (0.35 g) and EDCI (0.60 g) in methylene chloride was stirred at ambient temperature for 18 h. The solvent was removed in vacuo and the residue was dissolved in ethyl acetate and washed with water and brine. The solvent was dried (MgSO₄) and evaporated to a slurry which was triturated with ethyl acetate and hexane to yield 0.41 g of a white solid. MS(CI/NH₃) m/e 292 (M+H)⁺ ¹H NMR (CDCl₃,300MHz) δ 2.82 (br s,4H), 6.97-7.09 (m,2H), 7.30-7.41 (m,2H)

Example 170

BOC-Trp-Lys (epsilon-N-(5-hydroxyindole-2-carbonyl))- Asp-Phe-NH₂

The tetrapeptide in example 6 (100 mg), active ester in example 169 (40 mg) and N-methylmorpholine were allowed to react under the conditions described in example 8. The final peptide was isolated in an identical manner to yield 44 mg of a white solid. MS(FAB+) m/e 853 (M+H⁺). ¹H

NMR (DMSO-d₆, 500MHz) δ 1.10-1.68 (m, 6H), 1.32 (br s,9H), 2.40-2.48 (m,1H), 2.53-2.64 (m, 1H), 2.81-2.98 (m, 2H) , 3.03-3.15 (m,2H), 3.18-3.28 (m, 2H), 4.25 (br s,2H), 4.35 (q, 1H), 4.48 (q,1H), 6.69 (d, 1H), 6.78 (d,2H), 6.87 (s,1H)H 6.95 (t,1H), 7.00-7.25 (m,9H), 7.31 (d, 1H), 7.38 (br s,2H), 7.58 (d, 1H) , 7.94 (br d,2H), 8.18 (d, 1H), 8.43 (br s,1H), 8.70 (s,1H). Anal calcd for C₄₄H₅₂N₈O₁₀ 2H₂O 1.5C₂H₄O₂: C 57.66, H 6.38, N 11.44; found: C 57.60, H 5.89, N 11.71.

Example 171

BOC-Trp-Lys (epsilon-N-(3-methylphenylaminocarbonyl))- Asp-Phe-NH₂

A solution of the tetrapeptide of example 6 (110 mg), 3-methylphenyl isocyanate (25 uL) and N-methylmorpholine (33 mg) in DMF (5 mL) was allowed to react as described in example 157. The product was isolated in a similar manner to yield 86 mg of a white solid. MS(FAB+) m/e 827 (M+H)⁺. ¹H NMR (DMSO-d₆, 500MHz) δ 1.08-1.66 (m, 6H) , 1.31 (br s,9H), 2.22 (s,3H), 2.32-2.39 (m, 1H) , 2.44-2.55 (m, 1H), 2.81-3.25 (m,8H), 4.16-4.29 (m,2H), 4.35 (m, 1H), 4.42 (br q, 1H), 6.65 (br d, J=7.5 Hz,1H), 6.77 (br d, 1H), 6.94 (br t,1H), 7.03-7.40 (m, 15H), 7.54 (br d, 1H) , 7.88 (br d, 1H), 7.98 (br d,1H), 8.09 (br d, 1H). Anal calcd for C₄₃H₅₄N₈O₉ C₂H₄O₂ 0.75H₂O: C 59.96, H 6.66, N 12.43; found: C 59.97, H 6.31, N 12.45.

Example 172

BOC-Trp-Lys (epsilon-N-(4-chlorophenylaminocarbonyl))-Asp-Phe-NH₂

A solution of tetrapeptide of example 6 (100 mg), N-methylmorpholine (40 uL), and 4-chlorophenyl isocyanate (40 mg) in DMF (5 mL) was reacted according to the procedure in example 157. The product was isolated under identical conditions to yield 38 mg of a white solid. MS(FAB+) m/e 847 (M+H)+. ¹H NMR(DMSO-d₆, 500MHz) δ 1.08-1.68 (m, 6H), 1.29 (br s,9H), 2.40-2.64 (m,2H), 2.80-3.16 (m, 8H), 4.22 (br s,2H), 4.37 (br m, 1H), 4.48 (br m, 1H) , 6.74 (br m, 1H), 6.95 (t,1H), 7.02-7.38 (m, 14H), 7.41 (d, 1H), 7.57 (br d, 1H) , 7.75 (br d, 1H), 7.88 (br s,1H), 8.14 (br s,1H).

Anal calc for C₄₂H₅₁N₈ Cl H₂O 0.5C₂H₄O₂ : C 57.68, H 6.19, N 12.51; found: C 57.67, H 5.99, N 12.43.

Example 173

BOC-Trp-Lys (epsilon-N-(2-chlorophenylaminocarbonyl))-Asp-Phe-NH₂

A solution of tetrapeptide of example 6 (100 mg), 2-chlorophenyl isocyanate (30 uL) and N-methylmorpholine (35 uL) in DMF (5 mL) was stirred at ambient temperature for 18 h. The product was purified as described in example 157 to yield 72 mg of a white solid. MS(FAB+) m/e 847 (M+H)+. ¹H NMR (DMSO-d₆, 500MHz) δ 1.10-166 (m, 6H) , 1.31 (br s, 9H), 2.41 (dd, J=6Hz, J=17Hz,1H), 2.63 (dd, 1H), 2.83-3.14 (m, 8H), 4.20-4.30 (m,2H), 4.38 (m, 1H), 4.51 (q, 1H), 6.77 (br d, 1H), 6.90-7.40 (m,14H), 7.58 (d, 1H), 7.86 (d,1H), 7.90 (d, 1H), 8.00 (s,1H), 8.14 (d,1H), 8.21 (d, 1H). Anal calcd for

C₄₂H₅₁N₈O₉Cl H₂O 0.5C₂H₄O₂: C 57.68, H 6.19, N 12.51;

found: C 57.68, H 6.03, N 12.51.

Example 174

BOC-Trp-Lys (epsilon-N-(1-naphthylaminocarbonyl))-Asp-Phe-NH₂

A solution of tetrapeptide of example 6 (100 mg), 1-naphthyl isocyanate (30 uL) and N-methylmorpholine (30 uL) in DMF (5 mL) was allowed to react as described in example 157. The final product was purified in a similar manner to yield 54 mg of a white solid. MS(FAB+) m/e 863 (M+H)⁺. ¹H NMR (DMSO-d₆, 500MHz) δ 1.10-1.68 (m, 6H), 1.30 (br s,9H), 2.39-2.60 (m,2H), 2.82-3.18 (m, 8H), 4.24 (br s,2H), 4.36 (m, 1H), 4.45 (m, 1H), 6.78 (br d, 1H), 6.91-7.68 (m, 18H), 7.84-8.05 (m, 3H), 8.17 (br s,1H). Anal calcd for

C₄₆H₅₄N₈O₉ C₂H₄O₂ 0.6H₂O: C 61.74, H 6.39, N 11.99; found: C 61.67, H 6.19, N 11.99.

Example 175

BOC-Trp-Lys (epsilon-N-(phenylaminocarbonyl))- Asp-Phe-NH₂

A solution of tetrapeptide of example 6 (100 mg), phenyl isocyanate (20 uL) and N-methylmorpholine (30 uL) in DMF (5 mL) was reacted in a similar manner as described in example 157. The product was purified under identical conditions to yield 47 mg of a white powder. MS (FAB+) m/e 813 (M+H)⁺. ¹H NMR (DMSO-d₆, 500MHz) δ 1.08-1.66 (m, 6H), 1.30 (br s,9H), 2.35-2.60 (m, 2H) , 2.81-3.14 (m, 6H), 4.17- 4.28 (br m,2H), 4.35 (m, 1H), 4.45 (m, 1H), 6.82 (q, 2H), 6.95 (t,1H), 7.04 (t,1H), 7.09-7.45 (m, 14H), 7.58 (d, 1H) , 7.86 (d,1H), 7.98 (br d,1H), 8.18 (br d, 1H). Anal calcd for

C₄₂H₅₂N₈O₉ H₂O: C 60.71, H 6.55, N 13.49; found C 60.76, H 6.50, N 13.14.

Example 176

BOC-Trp-Lys (epsilon-N-(cyclohexylaminocarbonyl))-Asp-Phe-NH₂

A solution of tetrapeptide of example 6 (100 mg), cyclohexyl isocyanate (25 uL) and N-methylmorpholine (33 uL) in DMF (5 mL) was allowed to react as described in example 157. The product was purified in a similar manner to that described to yield 44 mg of a white solid.

MS(FAB+) m/e 819 (M+H)⁺. ¹H NMR (DMSO-d₆, 300MHz) δ 1.00-1.76 (m,17H), 1.30 (br s, 9H), 2.42 (dd, J=7Hz, J=16Hz, 1H), 2.59 (dd, 1H), 2.81-2.95 (m, 4H), 3.01-3.12 (m, 2H), 4.22

(m,2H), 4.36 (m,1H), 4.48 (q, 1H) , 6.78 (d, 1H), 6.95

(t, J=7Hz, 1H), 7.04 (t, 1H), 7.08-7.38 (m,8H), 7.58 (d, 1H),

7.85-7.95 (m, 2H) , 8.21 (d, 1H), Anal calcd for C₄₂H₅₈N₈O₉

H₂O 0.75C₂H₄O₂: C 59.17, H 7.20, N 12.66; found: C 59.11,

H 6.83, N 12.82.

Example 177

BOC-Trp-Lys (epsilon-N-(3-chlorophenylaminocarbonyl))-Asp-Phe-NH₂

A solution of tetrapeptide of example 6 (100 mg), 3-chlorophenyl isocyanate (40 uL) and N-methylmorpholine (35 uL) in DMF (5 mL) was allowed to react under the conditions described in example 157. The final product was purified under identical conditions to yield 46 mg of a white powder. MS(FAB+) m/e 847 (M+H) + . 1H NMR (DMSO-d₆, 500MHz) δ 1.08-1.65 (m,6H), 1.30 (br s,9H), 2.44 (dd, 1H), 2.62 (dd,1H), 2.81-2.96 (m,2H), 3.00-3.15 (m,4H), 4.22 (br s,2H), 4.36 (br m,1H), 4.48 (q, 1H), 6.81 (d, 1H), 6.89

(m, 1H), 6.95 (t, J=5Hz, 1H), 7.04 (t,1H), 7.09-7.33 (m, 10H), 7.58 (d,1H), 7.67 (s,1H), 7.82 (d, 1H), 7.94 (d, 1H), 8.20 (d, 1H), 10.76 (br s,1H). Anal calcd for C₄₂H₅₁N₈O₉Cl H₂O 0.5C₂H₄O₂: C 57.68, H 6.19, N 12.51; found: C 57.63, H 5.84, N 12.48.

Example 178

BOC-Trp-Lys (epsilon-N-[3-methylphenylaminothiocarbonyl])-Asp-Phe-NH₂

A solution of tetrapeptide of example 6 (100 mg), 3-methylphenyl isothiocyanate (30 uL) and N-methylmorpholine (35 uL) in DMF (5 mL) was allowed to react as described in example 157. The product was purified in a similar manner to yield 42 mg of a white solid. MS (FAB+) m/e 865 (M+Na)⁺. ¹H NMR (DMSO-d₆, 500MHz) δ 1.12-1.68 (m, 6H), 1.33 (br s,9H), 2.24 (s,3H), 2.28-2.46 (m, 2H), 2.82-2.98 (m, 3H), 3.07-3.17 (m,3H), 4.18 (br s,1H), 4.27 (M, 1H), 4.33 (m, 1H) , 4.39 (m,1H), 6.81 (m, 2H), 6.94 (t,1H), 7.02-7.43 (m, 12H), 7.57 (d, 1H), 7.84 (br d,1H), 8.02 (br d, 1H), 8.11 (br d,1H). Anal calcd for C₄₃H₅₄N₈O₈S 2H₂O 2C₂H₄O₂ : C 56.50, H 6.65, N 11.28; found: C 56.23, H 6.10, N 11.68.

Example 179

BOC-Trp-Lys (epsilon-N-(t-butylaminocarbonyl))- Asp-Phe-NH₂

A solution of tetrapeptide of example 6 (100 mg), t-butyl isocyanate (30 uL) and N-methylmorpholine (30 uL) in DMF (5 mL) was allowed to react under the conditions described in example 157. The product was purified in a similar manner to yield 67 mg of a white solid. MS (FAB+) m/e 793 (M+H)⁺. ¹H NMR (DMSO-d₆, 300MHz) δ 1. 03-1 . 58

(m,6H), 1.20 (br s, 9H), 1.30 (br s,9H), 2.40

(dd, J=Hz, J=16Hz,1H), 2.58 (dd, 1H), 2.78-2.98 (m, 4H), 3.02-3.14 (m,2H), 4.15-4.29 (m, 2H), 4.35 (m, 1H), 4.47 (q, 1H), 6.82 (d,J=8Hz,1H), 6.91-7.49 (m, 10H), 7.58 (d, 1H), 7.90-8.02 (m,2H), 8.22 (d, 1H), 10.85 (br s,1H). Anal calcd for

C₄₀H₅₆N₈O₉ H₂O 0.75C₂H₄O₂: C 58.23, H 7.18, N 13.09;

found: C 58.21, H 6.87, N 13.20.

Example 180

BOC-Trp-Lys (epsilon-N-(2-methylphenylaminocarbonyl))-ASP-(NMe)Phe-NH₂

A solution of the tetrapeptide of example 104 (219 mg), 2-methylphenyl isocyanate (100 uL) and N-methylmorpholine (35 uL) in DMF was allowed to react as described in example 8. The product was purified under similar conditions to yield a white solid. MS (FAB+) m/e 841 (M+H)⁺. ¹H NMR (DMSO-d₆, 300MHz) δ 1.14-1.56 (m, 6H), 1.29 (br s,9H), 2.05-2.40 (m,2H), 2.15 (s,3H), 2.27 (s,3H), 2.67-3.29 (m,8H), 2.97 (s,3H), 4.20-4.39 (m, 2H), 4.71-5.27 (m,2H), 6.78-7.98 (m, 16H) , 8.24 (s,1H), 8.58 (br s,1H), 10.82 (br s,1H). Anal calcd for C₄₄H₅₆N₈O₉ C₂H₄O₂ : C 61.32, H 6.61, N 12.44; found: C 61.46, H 6.22, N 12.18.

Example 181

BOC-Trp-3,5-dimethylpyrazolide

To a 0°C solution of BOC-Trp (1.5 g), 3,5-dimethylpyrazole (0.57 g) and HOBT (0.80 g) in methylene chloride (40 mL) was added EDCI (1.04 g). The reaction was allowed to warm to ambient temperature and stirred for 18 h. The solvent was removed in vacuo, the residue dissolved in ethyl acetate and washed with 1 M phosphoric acid solution, saturated sodium bicarbonate solution then brine. After drying over sodium sulfate, the solvent was removed in vacuo and the residue chromatographed on silica (3:1, hexane :ethyl acetate) to yield 1.48 g of a white product. MS(DCI/NH₃) m/e 383 (M+H)⁺. ¹H NMR (CDCl₃, 300MHz) δ 1.41 (br s,9H), 2.29 (s,3H), 2.40 (s,3H), 3.30-3.49 (m, 2H), 5.28 (br d,1H), 5.86 (m, 1H), 5.99 (br s,1H), 6.97 (s,1H,) 7.04 (t, J=7 . 5 Hz, 1H) , 7 . 15 (t, 1H) , 7 . 28-7 . 38 (m, 2H) , 8 . 23 (br s , 1H) .

Example 182

BOC-Trpψ(CH₂NH) Lys(epsilon-N-Cbz)-ASP(OBn)-Phe-NH₂

To a -78°C solution of the pyrazolide of example 181 (300 mg) in tetrahydrofuran (20 mL) was added lithium aluminum hydride (49 mg) in five portions over a 20 minute period. The reaction was poured into 10% citric acid solution (100 mL) then extracted with ether. The combined ether extract was washed with saturated sodium bicarbonate solution then brine and dried over sodium sulfate. The solvent was evaporated in vacuo and the residue was dissolved in tetrahydrofuran (25 mL) to which the free base of example 4 (415 mg) was added. After 1 h, acetic acid (45 uL) and sodium cyanoborohydride (49 mg) were added and allowed to stand for 1 h. The reaction mixture was diluted with ethyl acetate, washed with saturated sodium

bicarbonate solution, then dried over sodium sulfate. The solvent was removed in vacuo and the residue

chromatographed (silica, 5% methanol in chloroform) to yield 220 mg of a white solid. MS (FAB+) m/e 904 (M+H)⁺. ¹H NMR (DMSO-d₆, 300MHz) δ 1.12-1.48 (m, 6H) , 1.33 (br s,9H), 2.28-3.03 (m, 12H) , 3.71 (br s,1H), 4.35-4.45 (m, 1H), 4.64 (br q,1H), 4.99 (s,2H), 5.03 (s,2H), 6.62 (br d, 1H), 6.91-7.40 (m,12H), 7.55 (d, 1H), 7.86 (br d, 1H), 8.20 (br d, 1H) .

Example 183

BOC-Trpψ(CH₂NH)Lys-Asp-Phe-NH₂

A solution of peptide of example 182 (155 mg) in acetic acid (10 mL) was hydrogenated in the presence of 10% Pd-C (75 mg) under one atmosphere hydrogen gas at ambient temperature. The catalyst was filtered and the solvent removed in vacuo to yield 111 mg of a white solid

MS (FAB) ⁺ m/e 680 (M+H)⁺.

Example 134

BOC-Trpψ(CH₂NH)Lys(epsilon-N-(4-hydroxycinnamoyl))- Asp-Phe-NH₂

A solution of peptide of example 183 (70 mg), active ester of example 15 (32 mg) and N-methylmorpholine (12 uL) in DMF (7 mL) was allowed to react as described in example 8. The product was purified under similar conditions to yield 36 mg of a white solid. MS(FAB+) m/e 826 (M+H)⁺ . ¹H NMR (DMSO-d₆, 300MHz) δ 1.12-1.49 (m, 6H), 1.34 (br s, 9H), 2.30-3.17 (m,12H), 3.70 (br m, 1H), 4.33-4.42 (m, 1H), 4.53 (q,1H), 6.41 (d, J=16Hz,1H), 6.62 (br d, J=8Hz, 1H), 6.88 (d, J=8Hz, 1H), 6.91-7.42 (m, 16H), 7.56 (br d, 1H), 7.88-8.01 (m, 2H), 8.14 (d, 1H), 9.83 (br s, 1H), 10.75 (br s,1H). Anal calcd for C₄₄H₅₅N₇O₉ 2H₂O: C 61.31, H 6.90, N 11.37;

found: C 61.00, H 6.53, N 11.10.

Example 185

BOC-Lys(epsilon-N-Cbz)-3,5-dimethylpyrazolide

BOC-Lys(epsilon-N-Cbz) (2.00 g), 3, 5-dimethylpyrazole (0.61 g), HOBT (0.85 g), and EDCI (1.11 g) were allowed to react as described in example 181 to yield 2.32 g of a white solid. MS(DCI/NH₃) m/e 458 (M+H)⁺. ¹H NMR (DMSO- d₆,300MHz) δ 1.13-1.77 (m, 6H), 2.18 (s,3H), 2.44 (s,3H), 2.99 (br s,2H), 5.00 (s,2H), 5.17 (br m, 1H), 6.20 (s,1H), 7.20-7.40 (m, 5H) .

Example 186

BOC-Trp-Lys (eosilon-N-Cbz)ψ(CH₂NH)ASP(OBn)-Phe-NH₂

The pyrazolide of example 185 (403 mg) was reacted with lithium aluminum hydride (52 mg) then condensed with the free base of example 2 (287 mg) followed by reaction with acetic acid (50 uL) and sodium cyanoborohydride (62 mg) as described in example 182 to yield 326 mg of a white solid. MS(FAB+) m/e 718 (M+H)⁺. ¹H NMR(DMSO-d₆, 300MHz) δ 1.10-1.40 (m,6H), 1.37 (br s,9H), 1.98 (br m, 1H), 2.09-2.24 (m,2H), 2.37 (dd, 1H), 2.56 (dd,1H), 2.75-3.07 (m, 3H), 4.43-4.52 (m,1H), 5.00 (s,2H), 5.06 (s,2H), 6.40 (br d, 1H), 7.08-7.41 (m, 15H), 8.14 (d, 1H).

Example 187

Lys (epsilon-N-Cbz)ψ(CH₂NH)ASP(OBn)-Phe-NH₂ 2HCl

The peptide of example 186 (250 mg) was stirred in a solution of hydrochloric acid (g) in acetic acid for 2 h. The solvent was removed in vacuo and the residue dissolved in water and lyopholyzed to yield 239 mg of a white solid. MS(FAB+) m/e 618 (M+H)⁺.

Example 188

BOC-Trp-Lys (epsilon-N-Cbz)ψ(CH₂NH)Asp(OBn)-Phe-NH₂ A solution of peptide of example 187 (150 mg), N-methylmorpholine (50 uL) and BOC-Trp N-hydroxysuccinimide ester (96 mg) in methylene chloride (15 mL) was stirred at ambient temperature for 18 h. The reaction mixture was diluted with methylene chloride then washed with water and dried over sodium sulfate. The solvent was removed in vacuo and the residue chromatographed (silica, 3% methanol in chloroform) to yield 152 mg of a white solid. MS(FAB+) m/e 905 (M+H)⁺. ¹H NMR (DMSO-d₆, 300MHz) δ 1.08-1.46

(m, 6H) , 1.29 (br s,9H), 2.28-2.62 (m,2H), 2.76-3.11 (m, 4H), 3.64 (br S,1H), 4.16 (br m, 1H), 4.49 (br m, 1H), 4.99

(s,2H), 5.02 (s,2H), 6.74 (br d, 1H), 6.92-7.66 (m,21H), 8.13 (br d, 1H).

Example IS?

BOC-Trp-Lysψ(CH₂NH)Asp-Phe-NH₂

The peptide of example 188 (135 mg) was reacted as described in example 183 to yield 82 mg of a white solid. MS(FAB+) m/e 680 (M+H)⁺.

Example 190

BOC-Trp-Lys (epsilon-N-(4-hydroxycinnamoyl)) ψ(CH₂NH)Asp-Phe-NH₂

The peptide of example 189 (71 mg), N-methylmorpholine (13 uL) and active ester of example 15 (33 mg) were reacted as described in example 8 to yield, after a similar

purification procedure, 56 mg of a white solid. MS (FAB+) m/e 826 (M+H)⁺. ¹H NMR (DMSO-d₆, 300MHz) δ 1.09-1.50

(m,6H), 1.30 (br s,9H), 2.09-2.48 (m,3H), 2.73-3.19 (m, 8H), 3.68 (br s,1H), 4.16 (br s,1H), 4.48 (m, 1H), 6.42

(d, J=16Hz,1H), 6.68-6.79 (m,2H), 6.93-7.68 (m, 15H), 7.96 (br t,1H), 8.27 (br d,1H), 9.83 (br s,1H), 10.79 (s,1H). Anal calcd for C₄₄H₅₅N₇O₉ 2H₂O: C 61.31, H 6.90, N 11.37; found: C 61.33, H 6.53, N 11.28. Example 191

BOC-Trp-Lys (ε-N-[2-nitrophenylaminocarbonyl])-Asp-Phe-NH₂

The tetrapeptide of example 6, 2-nitrophenyl isocyanate and N-methymorpholine in DMF were allowed to react in a manner

described in example 157. Similar purification yielded the product as a white solid. MS (FAB+) m/e 858 (M+H)⁺. ¹H NMR(DMSO- d₆,300MHz) δ 1.08-1.65 (m,8H), 1.30 (s,9H), 2.41-2.53 (d, 1H), 2.66

(dd,1H), 2.70-3.13 (m, 6H), 4.16-4.32 (br s,2H), 4.33-4.43 (m, 1H), 4.46-4.55 (m, 1H), 6.84 (d, 1H) , 6.95 (t,1H), 7.02-7.34 (m, 9H), 7.55-7.65 (m,2H), 7.86 (d,1H), 7.95 (d, 1H), 8.03 (d,1H), 8.28 (t, 1H) . Anal calcd for C₄₂H₅₁N₉O₁₁ C₂H₄O₂ : C 57.57, H 6.04, N

13.73; found: C 57.93, H 5.90, N 13.90.

Example 192

BOC-Trp-Lys (ε-N-[2-triflurormethylphenylaminocarbonyl])- Asp-Phe-NH₂

The tetrapeptide of example 6 (175 mg, 0.26 mmol), N-methylmorpholine (0.06 mL) and 2-triflurormethylphenyl isocyanate (0.05 mL, 0.28 mmol) in DMF (5 mL) were allowed to react in a manner described in example 157. Similar purification yielded 135 mg of a white solid. MS (FAB+) m/e 881 (M+H)⁺. ¹H NMR (DMSO-d₆, 500MHz) δ 1.09-1.66 (m, 8H), 1.31 (s, 9H), 2.47 (dd, 1H), 2.65

(dd,1H), 2.82-2.96 (m, 2H), 2.99-3.13 (m, 4H), 4.19-4.32 (m,2H), 4.34-4.40 (m,1H), 4.48-4.54 (m, 1H) , 6.80 (d, 1H), 6.95 (t,1H), 7.02-7.33 (m,9H), 7.51-7.61 (m,2H), 7.27 (s,1H), 7.92 (d, 1H), 7.96

(d,1H), 8.25 (d,1H). Anal calcd for C₄₃H₅₁F₃N₈O₉ : C 58.63, H 5.84, N 12.72; found: C 58.26, H 5.81, N 12.46.

Example 193

BOC-Trp-Lys (ε-N-[2-hromophenylaminocarbonyl])-Asp-Phe-NH₂

A solution of the tetrapeptide of example 6 (175 mg, 0.26 mmol), N-methylmorpholine (0.06 mL, 0.52 mmol) and 2-bromophenyl isocyanate (0.034 mL, 0.28 mmol) in DMF (5 mL) was allowed to react as described in example 157 and the product purified in a similar manner to yield 130 mg of a white solid. MS (FAB+) m/e 893 (M+H)⁺. ¹H NMR (DMSO-d₆, 500MHz) δ 1.07-1.66 (m, 8H), 1.30 (s,9H),

2.44 (dd, 1H), 2.61 (dd, 1H), 2.82-2.95 (m,2H), 3.01-3.13 (m, 4H), 4.19-4.30 (m,2H), 4.33-4.39 (m, 1H), 4.49 (q, 1H), 6.79 (d, 1H), 6.95 (t,1H), 7.02-7.39 (m, 8H), 7.53 (d, 1H), 7.58 (d, 1H), 7.88 (s,1H), 7.93 (d,1H), 8.06 (d, 1H), 8.22 (d, 1H). Anal calcd for C₄₂H₅₁BrN₈O₉ H₂O: C 55.44, H 5.99, N 12.32; found: C 55.70, H 5.70, N 12.16

Example 194

BOC-Trp-Lys (ε-N-[2-chlorophenylaminothiocarbonyl])-Asp- Phe-NH₂

A solution of the tetrapeptide of example 6 (160 mg, 0.23 mmol), N-methylmorpholine (0.06 mL, 0.52 mmol) and 2-chloropheny isothiocyanate (0.05 mL, 0.26 mmol) in DMF (5 mL) was allowed to react as described in example 157. A similar purification yield 138 mg of a white solid. MS(FAB+) m/e 863 (M+H)⁺ . ¹H NMR(DMSO-d₆,300MHz) δ 1.09-1.65 (m,8H), 1.30 (s, 9H), 2.44 (dd, 1H), 2.63

(dd,1H), 2.80-2.96 (m, 2H), 3.00-3.18 (m, 4H), 4.17-4.30 (m,2H), 4.31-4.40 (m,1H), 4.45-4.53 (m, 1H), 6.84 (d, 1H), 6.95 (t,1H), 7.11-7.34 (m,8H), 7.46 (d, 1H), 7.59 (d, 1H), 7.67 (d, 1H), 7.87

(d,1H), 7.98 (d,1H). Anal calcd for C₄₂H₅₁ClN₈O₈S H₂O: C 57.23, 6.06, N 12.71; found: C 57.42, H 5.87, N 12.63.

Example 195

BOC-Trp-Lys (ε-N-[2-methylphenylaminothiocarbonyl])-Asp- Phe-NH₂

A solution of the tetrapeptide of example 6 (160 mg, 0.23 mmol), N-methylmorpholine (0.06 mL, 0.52 mmol) and 2-methylpheny isothiocyanate (0.04 mL, 0.26 mmol) in DMF (5 mL) was allowed to react in a manner similar to that described in example 157.

Purification of the mixture yielded 54 mg of a white solid.

MS(FAB+) m/e 843 (M+H)⁺ . ¹H NMR(DMSO-d6, 300MHz) δ 1.09-1.67

(m,8H), 1.30 (br s,9H), 2.44-2.66 (m,2H), 2.78-2.97 (m, 2H), 3.01-3.19 (m,2H), 4.16-4.29 (m, 2H), 4.30-4.39 (m, 1H), 4.42-4.54 (m, 1H), 6.82 (d,1H), 6.95 (t,1H), 7.10-7.37 (m, 7H), 7.58 (d, 1H), 7.87 (d, 1H), 7.98 (d,1H), 8.22 (d, 1H). Anal calcd for C₄₃H₅₄N₈O₈S H₂O: C 59.99, H 6.32, N 13.02; found: C 60.15, H 6.34, N 12.96.

Example 196

BOC-Trp-Lys (ε-N-[3-acetylphenylaminocarbonyl])-Asp- Phe-NH₂

The tetrapeptide of example 6 (160 mg, 0.23 mmol), N-methylmorpholine (0.06 mL, 0.52 mmol) and 3-acetylphenyl

isocyanate (0.05 mL, 0.26 mmol) were allowed to react in DMF (5 mL) as described in example 157. A similar purification yielded 96 mg of a white solid. MS(FAB+) m/e 855 (M+H)⁺. ¹H NMR(DMSO-d₆,300MHz) δ 1.08-1.66 (m, 8H), 1.31 (br s,9H), 2.52 (s,3H), 2.39- 2.66 (m,2H), 2.79-2.96 (m,2H), 2.98-3.15 (m, 4H), 4.18-4.28 (m, 2H), 4.31-4.40 (m,1H), 4.43-4.53 (m, 1H) , 6.85 (d, 1H), 6.95 (t, 1H), 7.05 (t,1H), 7.11-7.41 (m, 9H), 7.47 (d, 1H), 7.55-7.66 (m,2H), 7.86 (d,1H), 7.95-8.04 (m, 2H), 8.24 (d,1H). Anal calcd for C₄₄H₅₄N₈O₁₀ H₂O: C 60.54, H 6.47, N 12.84; found: C 60.32, H 6.28, N 12.64. Example 197

BOC-Trp-Lys (ε-N-[4-acetylphenylaminocarbonyl])-Asp- Phe-NH₂

A solution of the tetrapeptide of example 6 (160 mg, 0.23 mmol), N-methylmorpholine (0.06 mL, 0.52 mmol) and 4-acetylphenyl isocyanate (0.05 mL, 0.26 mmol) in DMF (5 mL) was allowed to react as described in example 157. The product was purified in an identical manner to yield 125 mg of a white solid. MS(FAB+) m/e 855 (M+H)⁺. ¹H NMR (DMSO-d₆, 300MHz) δ 1.09-1.65 (m, 8H), 1.30 (br s,9H), 2.33-2.67 (m,2H), 2.47 (s,3H), 2.80-2.95 (m, 2H), 3.01-3.14 (m,4H), 4.17-4.28 (m, 2H), 4.32-4.40 (m, 1H), 4.43-4.53 (m,1H), 6.85 (d,1H), 6.95 (t,1H), 7.05 (t,1H), 7.10-7.35 (m, 8H) , 7.52 (d, 1H), 7.58 (d,1H), 7.79-7.87 (m,2H), 7.99 (d, 1H). Anal calcd for

C₄₄H₅₄N₈O₁₀ 1.25H₂O: C 60.22, H 6.46, N 12.77; found: C 60.07, H 6.24, N 12.62.

Example 198

BOC-Trp-Lys (ε-N-[4-phenoxyphenylaminocarbonyl)]-Asp- Phe-NH₂

A solution of the tetrapeptide of example 6 (160 mg, 0.23 mmol), N-methylmorpholine (0.06 mL, 0.52 mmol) and 4-phenoxyphenyl isocyanate (0.06 mL, 0.26 mmol) in DMF (5 mL) was reacted

according to the procedure in example 157. Purification of the product yielded 135 mg of a white solid. MS (FAB+) m/e 905 (M+H)⁺. ¹H NMR (DMSO-d6, 300MHz) δ 1.09-1.66 (m, 8H), 1.30 (s, 9H), 2.26-2.68

(m,2H), 2.80-2.96 (m, 2H), 2.99-3.15 (m,4H), 4.19-4.29 (m,2H), 4.32-4.40 (m,1H), 4.45-4.53 (m, 1H), 6.83-6.99 (m, 4H), 7.01-7.09 (m,3H), 7.11-7.25 (m, 5H) , 7.28-7.43 (m, 4H) , 7.58 (d, 1H) . Anal calcd for C₄₈H₅₅N₈O₁₀ H₂O: C 62.46, H 6.33, N 12.14; found: C 62.66, H 6.22, N 11.96. Example 199

BOC-Trp-Lys (ε-N-[2-isopropylphenylaminocarbonyl])-Asp- Phe-NH₂

A solution of the tetrapeptide of Example 6 (160 mg, 0.23 mmol), 2-isopropylphenyl isocyanante (0.042 mL) and N-methylmorpholine in DMF was allowed to react as described in

Example 157. The product was isolated in a similar manner to yield 60 mg of a white solid. MS(FAB+) m/e 855 (M+H)⁺. ¹H

NMR (DMSO-d₆, 300MHz) δ 1.09-1.65 (m,8H), 1.14 (d, J=7Hz, 6H), 1.30

(s,9H), 2.78-3.17 (m, 6H), 4.18-4.29 (m, 2H), 4.31-4.40 (m, 1H), 4.42-4.51 (m,1H), 6.83 (d, 1H), 6.91-7.33 (m, 8H), 7.40 (br s,1H), 7.55-7.66 (m,2H), 7.97 (t,1H), 8.22 (d, 1H). Anal calcd for

C₄₅H₅₈N₈O₉ H₂O 0.75C₂H₄O₂: C 60.90, H 6.91, N 12.25; found: C 60.90, H 6.76, N 12.30.

Example 200

BOC-Trp-Lys (ε-N-TS-2(1-naphthyl)ethylaminocarbonyl])- Asp-Phe-NH₂

The tetrapeptide of example 6 (160 mg, 0.23 mmol) was allowed to react with α-(1-naphthyl)-ethyl isocyanate and N-methylmorpholine in DMF in a similar manner as described in example 157. After purification, the product was isolated as a white solid. MS (FAB+) m/e 892 (M+H) ⁺. ¹H NMR(DMSO-d₆, 500MHz) δ

1.18-1.68 (m, 6H), 1.36 (s, 9H, 1.50 (d,3H), 2.45-2.72 (m, 2H), 2.88-3.20 (m,4H), 4.25-4.33 (m,2H), 4.39-4.45 (m, 1H), 4.51-4.56 (m, 1H), 5.58-5.65 (m,1H), 6.85 (d, 1H), 7.02 (t,1H), 7.10 (t,1H), 7.15-7.39 (m, 6H), 7.51-7.66 (m, 5H), 7.85 (d, 1H), 7.94-8.00 (m,2H), 8.19 (d,1H). Anal calcd for C₄₈H₅₈N₈O₉ 3H₂O: C 61.00, H 6.83, N 11.76; found: C 60.94, H 6.61, N 11.31. Example 201

BOC-Trp-Lys (ε-N-[4-methylphenylaminocarbonyl])-Asp-Phe-NH₂

A solution of the tetrapeptide of Example 6 (100 mg, 0.15 mmol), N-methylmorpholine (0.035 mL) and 4-methylphenyl isocyanate (0.08 mL) in DMF was reacted under the conditions described in Example 157 and isolated to yield 31 mg of a white solid.

MS(FAB+) m/e (M+H)⁺. ¹H NMR (DMSO-d₆, 500MHz) δ 1.11-1.66 (m, 8H),

1.32 (s,9H), 2.19 (s,3H), 2.34-2.43 (m, 1H), 2.47-2.59 (m, 1H), 2.82-3.15 (m,4H), 4.17-4.27 (m,2H), 4.32-4.39 (m, 1H). 4.41-4.47 (m, 1H), 6.79 (d, 1H), 6.92-7.33 (m, 10H), 7.57 (d, 1H), 7.85 (d, 1H), 7.97 (d,1H), 8.15 (d, 1H). Anal calcd for C₄₃H₅₄N₈O₉ C₂H₄O₂ 1.75H₂O: C 58.90, H 6.74 N.12.21; found: C 58.80, H 6.36, N 12.44.

Example 202

BOC-Trp-Lys (ε-N-[2-methoxyphenylaminocarbonyl])-Asp- Phe-NH₂

A solution of the tetrapeptide of Example 6 (100 mg, 0.15 mmol), N-methylmorpholine (0.35 mL) and 2-methoxyphenyl isocyanate (0.05 mL) in DMF was allowed to react as described in Example 157. Purification yielded 47 mg of a white solid. MS (FAB+) m/e 843 (M+H)⁺. ¹H NMR (DMSO-d₆, 500MHz) δ 1.11-1.66 (m,8H),1.31 (s, 9H),

2.44 (dd,1H), 2.61 (dd, 1H), 2.81-2.95 (m, 2H), 2.99-3.14 (m, 3H), 3.79 (s,3H), 4.19-4.31 (m, 2H) , 4.33-4.40 (m, 1H), 4.47-4.53 (m,1H), 6.76-6.88 (m,3H), 6.91-6.99 (m,2H), 7.04 (t,1H), 7.08-7.41 (m, 5H), 7.58 (d,1H), 7.85-7.96 (m,2H), 8.08 (dd, 1H), 8.22 (d,1H). Anal calcd for C₄₃H₅₄N₈O₈ 0.5C₂H₄O₂: C 60.68, H 6.46, N 12.92; found: C 60.85, H 6.60, N 12.84.

Example 203

BOC-Trp-Lys (ε-N-[2-naphthyl])-Asp-Phe-NH₂

A solution of the tetrapeptide of example 6 (100 mg, 0.14 mmol), N-methylmorpholine and 2-naphthyl isocyanate (0.03 mL) was allowed to react as described in example 157. Purification yielded 50 mg of a white solid. MS (FAB+) m/e 863 (M+H)⁺. ¹H NMR (DMSO-d₆, 500MHz) δ 1.07-1.49 (m, 8H), 1.30 (s,9H), 2.45 (dd, 1H),

2.64 (dd, 1H), 2.78-3.17 (m, 4H), 4.17-4.30 (m,2H), 4.31-4.41

(m, 1H), 4.40-4.55 (m, 1H), 6.87 (d, 1H), 6.95 (t,1H), 7.05 (t,1H), 7.11-7.46 (m, 10H), 7.59 (m, 1H),. Anal calcd for C₄₆H₅₆N₈O₉ H2O: C 62.71, H 6.41, N 12.72; found: C 62.34, H 6.21, N 12.35.

Example 204

BOC-Trp-Lys (ε-N-[ 2-(methoxycarbonyl)phenylaminocarbonyl])-Asp- Phe-NH₂

A solution of the peptide of example 6 (100 mg, 0.14 mmol), 2-(methoxycarbonyl)phenyl isocyanate (0.03 mL) and N-methylmorpholine in DMF was allowed to react as described in example 157. The product was purified in a similar manner to yield 50 mg of a white solid. MS (FAB+) m/e 871 (M+H)⁺ . ¹H

NMR (DMSO-d₆, 300MHz) δ 1.08-1.66 (m, 8H) , 1.30 (s,9H), 2.45 (dd, 1H),

2.64 (dd,1H), 2.79-3.14 (m, 4H) , 3.83 (s,3H), 4.16-4.31 (m,2H), 4.32-4.42 (m, 1H), 4.46-4.57 (m, 1H), 6.83 (d, 1H), 6.91-7.38

(m,10H), 7.44-7.53 (m,3H), 7.58 (d, 1H), 7.86-7.98 (m,3), 8.28 (d,1H), 8.37 (d,1H). Anal calcd for C₄₄H₅₄N₈O₁₁ 0.5H₂O 0.5C₂H₄O₂: C 59.52, H 6.31, N 12.40; found: C 59.54, H 6.24, N 12.39.

Example 205

BOC-Trp-Lys (ε-N-13-(methoxycarbonyl)phenylaminocarbonyl])-Asp- NH₂

The procedure described for Example 203 using

3-(methoxycarbonyl)phenyl isocyanate in place of

2-(methoxycarbonyl)phenyl isocyanate yielded the product as a white solid. MS (FAB+) m/e 893 (M+Na)⁺. ¹H NMR (DMSO-d₆, 300MHz) δ

1.08-1.46 (m,8H), 1.30 (s, 9H), 2.43 (dd, 1H), 2.62 (dd, 1H), 2.78-3.83 (m,6H), 3.82 (s,3H), 4.18-4.28 (m, 2H), 4.31-4.39 (m, 1H), 4.41-4.53 (m,1H), 6.87 (d, 1H), 6.95 (t,1H), 7.04 (t,1H), 7.10-7.38 (m,10H), 7.46 (d, 1H), 7.59 (d,2H), 7.36 (d,1H), 8.00 (d, 1H). Anal calcd for C₄₄H₅₄N₈O₁₁ H₂O 0.75C₂H₄O₂: C 58.45, H 6.37, N 11.96; found: C 58.59, H 6.04, N 12.28.

Example 206

BOC-Trp-Lys (ε-N-[2,6-dichlorophenylaminocarbonyl])- Asp-Phe-NH₂

A solution of the tetrapeptide of example 6 (100 mg), 2,6-dichlorophenyl isocyanate and N-methylmorpholine in DMF was allowed to react as described in Example 157. The product was purified to yield 66 mg of a white solid. MS(FAB+) m/e 881 (M+H)⁺ ¹H NMR (DMSO-d₆, 300MHz) δ 1.10-1.47 (m, 8H), 1.31 (s,9H), 2.42

(dd,1H), 2.60 (dd,1H), 2.79-3.17 (m,4H), 4.14-4.29 (m,2H), 4.31-4.41 (m,1H), 4.43-4.52 (m, 1H) , 6.81 (d, 1H), 6.95 (t,1H), 7.04 (t,1H), 7.10-7.38 (m,8H), 7.47 (d, 1H), 7.57 (d, 1H), 7.86-7.98

(m,3H), 8.22 (d, 1H). Anal calc for C₄₂H₅₀N₈O₉CI₂ H₂O: C 56.06, H 5.83, N 12.45; found: C 56.04, H 5.69, N 12.11.

Example 207

BOC-Trp-Lys (ε-N-[2 ,6-dimethylphenylaminocarbonyl])- Asp-Phe-NH₂

The tetrapeptide of Example 6 (100 mg), 2,6-dimethylphenyl isocyanate and N-methylmorphline were allowed to react in DMF as described in Example 157. The product was chromatographed to yield 73 mg of a white solid. MS(FAB+) m/e 841 (M+H)⁺. ¹H

NMR (DMSO-d₆, 300MHz) δ 1.08-1.48 (m, 8H), 1.30 (s,9H), 2.15 (s, 6H),

2.44 (dd, 1H), 2.63 (dd,1H), 2.79-3.16 (m, 4H), 4.17-4.29 (m, 2H), 4.31-4.41 (m,1H), 4.45-4.55 (m,1H), 6.83 (d, 1H), 6.92-7.35

(m, 12H), 7.58 (d, 1H), 7.88 (d, 1H), 7.95 (d, 1H), 8.26 (d, 1H). Anal calcd for C₄₄H₅₆N₈O₉ H₂O: C 61.52, H 6.80, N 13.04; found: C

61.69, H 6.53, N 12.71.

Example 208

BOC-Trp-Lys (ε-N-[allylaminocarbonyl])-Asp-Phe-NH₂

A solution of the tetrapeptide of Example 6 (100 mg, 0.15 mmol), allyl isocyanate (0.015 mL) and N-methylmorpholine (0.03 mL) in DMF was allowed to react according to the procedure in

Example 157. The product was isolated in a similar manner to yield 56 mg of a white solid. MS(FAB+) m/e 777 (M+H)⁺. ¹H

NMR (DMSO-d₆, 300MHz) δ 1.08-1.42 (m, 8H), 1.30 (s,9H), 2.38 (dd, 1H),

2.55 (dd, 1H), 2.78-3,16 (m, 4H), 3.62 (br m,2H), 4.16-4.28 (m, 2H), 4.30-4.39 (m, 1H), 4.41-4.49 (m, 1H), 4.99 (dm, 1H), 5.09 (dm, 1H), 7.05 (t,1H), 7.09-7.35 (m, 8H), 7.58 (d, 1H), 7.94 (d, 1H), 8.02 (d,1H), 8.21 (d,1H). Anal calcd for C₃₉H₅₂N₈O₉ 1.5H₂O: C 58.40, H 6.88, N 13.97; found: C 58.11, H 6.53, N 13.76.

Example 209

BOC-Trp-Lys (ε-N-[4-nitrophenylaminocarbonyl])-Asp-Phe-NH₂

A solution of the tetrapeptide of Example 6 (100 mg), 4-nitrophenyl isocyanate (24 mg) and N-methylmorpholine (0.03 mL) was allowed to react as described in Example 157. The reaction was chromatographed to yield 70 mg of a white solid. MS(FAB+) m/e 858 (M+H)⁺. ¹H NMR (DMSO-d₆, 300MHz) δ 1.05-1.69 (m, 8H), 1.29

(s,9H), 2.43 (dd,1H), 2.61 (dd, 1H), 2.80-3.16 (m, 4H), 4.15-4.28 (m,2H), 4.31-4.40 (m, 1H), 4.42-4.53 (m, 1H), 6.86 (d, 1H), 6.95 (t,1H), 7.04 (t,1H), 7.10-7.35 (m, 10H), 7.58 (d, 1H), 7.62 (d, 1H), 7.84 (d,1H), 8.00 (d, 1H), 8.10 (d,1H), 8.20 (d, 1H). Anal calcd for C₄₂H₅₁N₉O₁₁ 0.5H₂O 0.5C₂H₄O₂: C 57.47, H 6.08, N 14.03; found: C 57.48, H 5.94, N 14.14.

Example 210

BOC-Trp-Lys (ε-N-[benzylaminocarbonyl])-Asp-Phe-NH₂

A solution of the tetrapeptide of Example 6 (100 mg), benzyl isocyanate (0.02 mL) and N-methylmorpholine (0.03 mL) in DMF was reacted according to the procedure in Example 157. After

chromatography, the product was isolated as a white solid.

MS(FAB+) m/e 827 (M+H)⁺. ¹H NMR (DMSO-d₆, 30OMHz) δ 1.08-1.61

(m, 8H), 1.30 (s, 9H), 2.78-3.15 (m, 8H) , 4.15-4.28 (m, 4H), 4.31-4.40 (m, 1H), 4.43-4.52 (m, 1H), 6.83 (d,1H), 6.95 (t,1H), 7.04 (t,1H), 7.10-7.38 (m, 8H), 7.58 (d, 1H), 8.25 (d, 1H). Anal calcd for

C₄₃H₅₄N₈O₉ H₂O: C 61.12, H 6.68, N 13.26; found: C 61.33, H 6.39, N 13.03.

Example 211

BOC-Trp-Lys (e-N-[2-methylphenylaminocarhonyl])- Asp-HPhe-NH₂ a. BOC-Homophenylalanine amide

A solution of homophenylalanine (95 mg), triethylamine (0.18 mL), and di-t-butyl dicarbonate (180 mg) in 10 mL water and 10 mL acetone was stirred at room temperature overnight. The acetone was removed in vacuo, the aqueous phase diluted with water and extracted with ethyl acetate three times. The organic phase was washed with IM phosphoric acid (3 times), saturated sodium

bicarbonate solution (3 times), brine and dried over magnesium sulfate. The solvent was removed in vacuo to yield 127 mg (86%) of a viscous oil. The oil (127 mg) was dissolved in THF (15 mL) and cooled to -15° C to which N-methylmorpholine (0.051 mL) and isobutylchloroformate (0.60 mL) were added and stirred for 5 min at -10° C. A solution of ammonium hydroxide (2 mL) was then added, the solution stirred for 15 minutes and then allowed to warm to ambient temperature. After 4h, the reaction mixture was diluted with ethyl acetate, washed successively with 1M phosphoric acid, saturated sodium bicarbonate solution and brine. After drying over magnesium sulfate, the solvent was removed in vacuo to yield 116 mg of a white solid. ¹H NMR(CDCl₃,300MHz) δ 1.46 (s,9H), 1.87- 2.00 (m,1H), 2.13-2.28 (m, 1H), 2.72 (t, J=7Hz, 2H), 4.05-4.16

(m, 1H), 7.16-7.33 (m, 5H). b . HCl Asp(OBn)-HPhe-NH₂

A solution of the compound of example 211a (112 mg) in 10 mL HCl in acetic acid was stirred at ambient temperature for 2 h.

The product was precipitated with addition of ether, collected and dried. The salt (35 mg) was then coupled with BOC-Asp (OBn) activated with isobutylchloroformate in the presence of N- methylmorpholine. A similar work-up as described in Example 211a yielded the dipeptide which was deprotected using HCl in acetic acid to yield the hydrochloride salt. MS(CDI/NH₃) m/e 384 (M+H)⁺. 1H NMR (DMSO-d₆, 300MHz) δ 1.77-1.91 (m, 1H), 1.93-2.03 (m, 1H), 2.55- 2.69 (m,2H), 2.88 (dd, 1H), 3.07 (dd, 1H), 4.19-4.30 (m, 1H), 5.17 (s,2H), 7.14-7.47 (m, 5H). c . BOC-Trp-Lys (ε-N- [2-methylphenylaminocarbonyn ) - Asp-HPhe-NH₂

To a -10° C solution of BOC-Trp-Lys (ε-N-[2-methylphenylaminocarbonyl]) (99 mg) in 10 mL THF were added N-methyl morpholine (0.02 mL) and isobutylchloroformate (0.024 mL). The heterogeneous suspension was stirred for 3 minutes and a solution of compound of Example 211b (70 mg) and N-methylmorphline (0.02 mL) in THF (5 mL) was added. After stirring for 15 minutes at -10° C, the reaction was warmed to ambient temperature, allowed to stand for 4 h and worked up by dilution with ethyl acetate and washing successively with IM phosphoric acid, saturated sodium bicarbonate solution and brine followed by drying over magnesium sulfate. After solvent evaporation, the residue was

chromatographed on silica gel using ethyl

acetate:pyridine:water:acetic acid to yield a white solid after lyopholization. The peptide was then treated with 10% Pd-C in acetic acid under a balloon of hydrogen gas overnight. The catalyst was filtered and washed with fresh acetic acid and evaporated to a residue that was purified on silica gel using ethyl acetate:pyridine:acetic acid:water to yield a white solid after lyopholization. MS (FAB+) m/e 841 (M+H)⁺. ¹H NMR (DMSO-d₆, 300MHz) δ 1.11-1.42 (m, 6H), 1.30 (s, 9H), 1.49-1.72 (m, 2H), 1.77- 1.89 (m,1H), 1.94-2.06 (m, 2H), 2.30 (dd, 1H), 4.02-4.13 (m, 1H), 4.18-4.34 (m,2H), 4.50-4.58 (m, 1H), 6.82 (t, 1H), 6.94 (t,1H), 7.01-7.35 (m,10H), 7.58 (d, 1H), 7.81 (d, 1H) . Anal calcd for

C₄₄H₅₆N₈O₉ H₂O 1.5C₂H₄O₂: C 59.53, H 6.84, N 12.07; found: C 59.58, H, 6.38, N 12.47.

Example 212

BOC-Trp-Lys (ε-N-[2-methylphenylaminocarbonyl])- Asp-Phenylalaninol a. HCl Asp(OBn)-Phenylalaninol

A solution of BOC-Asp (OBn) (643 mg), phenylalaninol (300 mg), N-hydroxybenzotriazole (407 mg) and EDCI (456 mg) in methylene chloride was stirred at ambient temperature for 18 h. The mixture was washed with IM phosphoric acid, saturated sodium bicabonate solution and brine then dried over magnesium sulfate. Solvent evaporation yielded a white solid. The solid (300 mg) was dissolved in a 0° C solution of methylene chloride (20 mL) and trifluoroacetic acid (10 mL) and stirred for 3 h. The reaction was concentrated, dissolved in water and lyopholyzed to yield a white powder (288 mg). MS (DCI/NH₃) m/e 357 (M+H)⁺. ¹H

NMR (CDCI₃, 300MHz) d 2.62-2.81 (m, 2H), 2.85-3.00 (m, 2H), 3.40-3.53

(m,2H), 3.97-4.08 (m, 1H), 4.26-4.35 (m, 1H), 5.02 (s,2H), 7.08-7.38

(m, 10H), 7.83 (d, 1H). b_. BOC-Trp-Lys (ε-N-[2-methylphenylaminocarbonyl])- Asp-Phenylalaninol

A solution of BOC-Trp-Lys (ε-N-[2-methylphenylaminocarbonyl]) and the dipeptide salt of Example 212a was reacted as described in Example 211c to yield a white solid. MS(FAB-t-) m/e 814 (M+H)⁺. ¹H NMR (DMSO-d₆, 300MHz) δ 1.08-1.45 (m, 6H), 1.30 (s,9H), 2.15-2.70

(m, 3H), 2.75-3.18 (m, 6H), 3.78-3.91 (m, 1H), 4.18-4.28 (m,2H), 4.45-4.52 (m,1H), 6.80-6.90 (m, 1H) , 6.95 (t, 1H), 7.02-7.33

(m,10H), 7.59 (d, 1H), 8.00 (d, 1H). Anal calcd for C₄₃H₅₅N₇O₉ H₂O 0.5C₂H₄O₂: C 61.31, H 6.90, N 11.37; found: C 61.22, H 6.50, N 11.49.

Example 213

BOC-Trp-Lys (ε-N-[2-methylphenylaminocarbonyl])-Asp- NMe-Phenylalaninol

The product was prepared in a similar manner described for Example 212 beginning with available (NMe)phenylalaninol in place of phenylalaninol. MS(FAB+) m/e 828 (M+H)⁺. ¹H NMR (DMSO-d₆, 300MHz) δ 1.11-1.65 (m, 6H), 1.30 (s,9H), 2.15 (s,3H), 2.30

(dd,1H), 2.63-2.95 (m, 9H), 2.97-3.12 (m,2H), 3.37 (dd, 1H), 3.48 (dd,1H), 4.18-4.32 (m, 2H), 4.52 (m,0.5H), 4.63 (m,0.5H), 4.92-5.01 (m,1H), 6.78 (d, 1H), 6.84 (t,1H), 6.95 (q, 1H), 7.02-7.33 (m, 10H), 7.59 (d,1H), 7.78-7.88 (m,2H), 8.18-8.29 (m,2H). Anal calcd for C₄₄H₅₇N₇O₉ H₂O: C 62.47, H 7.03, N 11.59: found: C 62.22, H 6.75,

N 11.53.

Example 214

(Isobutoxycarbonyl)indolelactoyl-Lys[ε-N-(2-methylphenylaminocarboonyl])-Asp-Phe-NH₂

To a -10° C solution of indolelactic acid (100 mg) in 10 mL THF were added N-methylmorpholine (0.059 mL) and

isobutylchloroformate (0.07 mL) to form a heterogeneous suspension that was stirred for five minutes. To this mixture was added HCl Lys(Z)-Asp(OBn)-Phe-NH₂ (325 mg) and N-methylmorpholine (0.06 mL) in 2 mL DMF and stirred an additional 15 minutes at -10° C. The reaction was allowed to warm to ambient temperature and stirred for 4 h. The product was precipitated by addition of methylene chloride, collected then dissolved in acetic acid to which 10% Pd- C was added and stirred under a balloon of H₂ gas overnight. The catalyst was filtered and the acetic acid evaporated and the residue lyophαlyzed. The resulting peptide was then dissolved in DMF to which N-methylmorpholine and 2-methylphenyl isocyanate were added and allowed to stand overnight. The product was purified by chromatography in silica gel using ethyl acetate:pyridine:acetic acid:water to yield a white solid upon lyopholyzation. MS (FAB+) m/e 828 (M+H)⁺. ¹H NMR (DMSO-d₆, 500MHz) δ 0.80-0.85 (m, 6H), 1.08- 1.63 (m,6H), 1.75-1.85 (m, 1H), 2.15 (s,1.5H), 2.17 (s,1.5H), 2.38-2.66 (m,2H), 2.80-2.87 (m, 1H), 2.97-3.25 (m, 6H), 3.75-3.81 (m, 1H), 4.19-4.28 (m, 1H), 4.32-4.38 (m, 1H), 4.43-4.52 (m, 1H), 5.08-5.15 (m,1H), 6.83 (t,1H), 6.97 (t,1H), 7.03-7.26 (m, 1H), 7.32 (d, 1H), 7.61-7.65 (m, 1H), 7.78-7.88 (m, 2H). Anal calcd for C₄₃H₅₃N₇O₁₀ H₂O: C 61.05, H 6.55, N 11.59; found: C 61.05, H 6.38, N 11.59.

Example 215

Indolelactoyl-Lys(ε-N-[2-methylphenylaminocarbonyl])- Asp-Phe-NH₂

The compound was prepared in an identical manner described in Example 214 using benzylchloroformate instead of

isobutylchloroformate in the coupling of indolelactic acid with HCl Lys(Z)-Asp(OBn)-PheNH₂. MS (FAB+) m/e 728 (M+H)⁺. ¹H NMR(DMSO-d₆,500MHz) δ 1.00-1.65 (m, 6H), 2.17 (br s,3H), 2.38-2.48 (m, 1H),

2.61 (dd,1H), 2.78-3.36 (m, 6H), 4.15-4.20 (m, 1H), 4.22-4.28

(m,1H), 4.32-4.38 (m, 1H), 4.43-4.52 (m, 1H) , 6.85 (t,1H), 6.94 (q,1H), 7.01-7.39 (m, 9H), 7.53 (dd,1H), 7.59 (d, 1H), 7.72-7.92 (m,3H). Anal calcd for C₃₈H₄₅N₇O₈ H₂O: C 61.20, H 6.35, N 13.15; found: C 60.88, H 6.11, N 12.97.

Examole 216

BOC-Trp-Lys (ε-N-[4-hydroxycinnamoyl])-Asp-Trp-NH₂ a. HCl Asp(OBn)-Trp-NH₂

A solution of BOC-Asp(OBn) (377 mg), HCl Trp-NH₂ (277 mg), N-methylmorpholine (0.14 mL), N-hydroxybenzotriazole (234 mg) and EDCI (243 mg) in DMF (15 mL) was stirred at ambient temperature for 18 h. The reaction was diluted with ethyl acetate, washed successively with IM phosphoric acid, saturated sodium bicarbonate solution then brine. After drying over sodium sulfate, solvent evaporation yielded 0.56 g (96%) of a light brown solid. This dipeptide (425 mg) was then dissolved in 10 mL of HCl in acetic acid and stirred at ambient tmeperature for 2 h. Ether was added to precipitate the product which was collected, washed and dried to yield 300 mg (81%) of a light pink powder. MS (FAB+) m/e 409 (M+H)⁺. b. BOC-Trp-Lys (ε-N-[4-hydroxycinnamoyl])-Asp-Trp-NH₂

The dipeptide salt of Example 216a (200 mg) was coupled with BOC-Trp-Lys (Z) (255 mg) in the presence of N-hydroxybenzotriazole (91 mg), N-methylmorpholine (0.055 mL) and EDCI (94 mg) in

DMF/methylene chloride. Upon completion, the reaction was diluted with ethyl acetate and washed successively with 1M phosphoric acid, saturated bicarbonate solution and brine. After drying over sodium sulfate, the solvent was evaporated and the residue

chromatographed on silica gel using methanol:chloroform to yield 229 mg of the product as a white solid. The peptide (120 mg) was then dissolved in acetic acid to which 60 mg of 10% Pd-C was added then allowed to stir overnight under a balloon of hydrogen gas. The catalyst was filtered, washed with fresh acetic acid then evaporated to a residue which was dissolved in a minimum of methanol and the product precipitated with the addition of ether to collect 65 mg of a light pink solid. The product (60 mg) was dissolved in DMF (5 mL) to which N-methylmorpholine (0.01 mL) and 4-hydroxycinnamic acid N-hydroxysuccinimide ester (26 mg) were added and stirred at ambient temperature for 18 h. The DMF was removed in vacuo and the residue chromatographed on silica gel with ethyl acetate:pyridine:acetic acid:water to obtain the product as a white solid after lyopholyzation. MS (FAB+) m/e 879 (M+H)⁺. ¹H NMR (DMSO-d₆, 500MHz) δ 1.11-1.65 (m, 6H), 1.28 (s, 9H),

2.38-2.63 (m, 2H), 2.87-3.23 (m, 6H), 4.19-4.30 (m, 2H), 4.35-4.42 (m, 1H), 4.45-4.52 (m, 1H), 6.42 (d, J=16Hz, 1H), 6.68-6.78 (m, 3H),

6.92-7.18 (m, 7H), 7.29-7.43 (m, 5H), 7.55-7.60 (m, 2H), 7.90-7.99 (m,2H). Anal calcd for C₄₆H₅₄N₈O₁₀ 2.5H₂O: C 59.79, H 6.44, N

12.13; found: C 59.75, H 6.14, N 11.86.

Example 217

BOC-Trp-Orn (δ-N-[2-methylphenylaminocarbonyl])-Asp-Phe-NH₂

A solution of B0C-Trp-0rn-Asp-Phe-NH₂ (prepared in an

identical manner to that described for Example 6 by substituting BOC-Orn(N-δ-Cbz) for BOC-Lys (N-ε-Cbz)), N-methylmorpholine and 2-methylphenyl isocyanate in DMF was stirred at ambient temperature for 18 h. The DMF was removed in vacuo and the residue

chromatographed on silica gel using ethyl

acetate :pyridine:water:acetic acid to yield the product as a white solid after lyopholyzation. MS(FAB+) m/e 813 (M+H)⁺. ¹H NMR (DMSO-d₆, 300MHz) δ 1.09-1.72 (m, 4H) , 1.30 (s,9H), 2.17 (s,3H), 2.31

(dd, 1H), 2.45 (dd,1H), 2.75-3.18 (m, 6H), 4.16-4.47 (m, 4H), 6.73-6.88 (m,2H), 6.95 (t, 1H), 7.00-7.35 (m, 10H), 7.59 (d, 1H), 7.82 (d, 1H), 8.05 (d,1H), 8.13-8.26 (m,2H). Anal calcd for C₄₂H₅₂N₈O₉ 1 . 5H₂O : C 57 . 98 , H 6 . 64 , N 11 . 88 ; found : C 58 . 01 , H 6 . 26 , N 11 . 75 .

Example 218

BOC-Trp-Orn (δ-N-[4-hydroxycinnamoyl])-Asp-Phe-NH₂

The compound was prepared in a similar manner to that

described for Example 217 by substituting 4-hydroxycinnamic acid N-hydroxysuccinimide ester for 2-methylphenyl isocyanate.

MS(FAB+) m/e 826 (M+H)⁺. ¹H NMR (DMSO-d₆, 300MHz) δ 1.08-1.66

(m,4H), 1.30 (s,9H), 2.39 (dd, 1H), 2.78-2.95 (m,2H), 3.00-3.22 (m,4H), 4.15-4.26 (m, 1H), 4.27-4.40 (m,2H), 4.43-4.52 (m, 1H), 6.42 (d, J=16Hz,1H), 6.73-6.82 (m, 2H) , 6.95 (t,1H), 7.03 (t, 1H), 7.08-7.42 (m,8H), 7.59 (d, 1H), 7.92-8.28 (m,3H). Anal calcd for

C₄₃H₅₁N₇O₁₀ C₂H₄O₂ 3H₂O: C 57.50, H 6.54, N 10.43; found: C 57.44, H 5.85, N 10.77.

Example 219

BOC-Trp-HLys (ω-N-[2-methylphenylaminocarbonyl])-Asp- Phe-NH₂ a . Nα-Boc,Nω-(2-methylphenylaminocarbonyl)-L- Homolysine

N^α-Boc-3, 4-dehydro-L-homolysine (0.153 g, 0.59 mmol) prepare by a modification of the method of Schiller and

Beaulieu. Tetrahedron Lett. , 29 (17 ) , 2019 (1988) was dissolved in DMF (2ml) containing DIEA (0.115 ml, 0.65 mmol) and treated with o-tolyl isocyanate (0.085 ml, 0.67 mmol). The flask was capped with a drierite filled drying tube and the contents allowed to stir at room temperature overnight. The reaction mixture was subsequently partitioned between ethyl acetate and dilute aqueous HCl. The organic phase was dried (MgSO₄), filtered and concentrated in vacuo to give the crude product which was purified by flash chromatography on silica gel (ethyl acetate-hexane-acetic acid then ethyl acetate-acetone-acetic acid) to give 0.131 g (0.355 mmol) of an oil. MS (El) m/e 392 (M+H)⁺. The unsaturated product was dissolved in ethyl acetate and hydrogenated (10% Pd/C, 4 atm. H2) at room temperature. Filtration through celite and

concentration in vacuo gave 0.102 g (0.26 mmol) of an oil. b. Nα-Boc,Nω-(2-methylphenylaminocarbonyl)-Hlys-Asp-Phe-NH₂

The product from example 219a (0.098 g, 0.25 mmol) was coupled to Asρ-Phe-NH₂ (added as the hydrochloride, 0.08 g, 0.25 mmol) via standard mixed anhydride methodology. The reaction mixture was subsequently added dropwise to a large volume of dilute aqueous hydrochloric acid with vigorous agitation. The crude product was collected by vacuum filtration, water washed and dried in vacuo at room temperature to give 0.102 g (0.155 mmol) of a solid product sufficiently pure for further use as isolated. MS (FAB)+ m/e 655 (M+H)⁺ m/e 677 (M+Na)⁺. c. Boc-Trp-Hlys (Nω- [2-methylphenylaminocarbonyl])- Asp-Phe-NH₂

The product from example 219b (0.102 g, 0.155 mmol) was treated with 1.5N HCl in glacial acetic acid (7 mL) at room temperature in a tightly capped flask. After forty- five minutes the reaction mixture was frozen and lyophilized. The hydrochlorid (0.155 mmol) was combined with Boc-Trp-OSu (0.062 g, 0.155 mmol) in DMF (2 mL) containing DIEA (0.06 mL, 0.34 mmol) under nitrogen at room temperature and allowed to stir overnight. The reaction mixture was subsequently added dropwise to a large volume of dilute aqueous HCl and the crude product collected by vacuum filtration, water washed and dried in vacuo at room temperature. Purification by recrystallization from aqueous ethanol gave 0.110 g (0.131 mmol) of a granular off-white solid. MS (FAB+) m/e 841 (M+H)⁺ m/e 863 (M+Na)⁺. ¹H NMR (DMSO-d₆, 300MHz) δ 1.1-1.7 (m, 8H),

1.31 (s,9H), 2.4-3.15 (m, 8H), 4.15-4.45 (m,3H), 4.52 (m, 1H), 6.52 (t,1H), 6.8-7.35 (m,15H), 7.55-7.65 (m,2H), 7.8-8.0 (m, 3H), 8.29 (db, J=7.5Hz,1H), 10.8 (s,1H). Anal. calc. for C₄₄H₅₆N₈O₉·H₂O :

C, 61.51; H,6.82; N,13.05. Found: C,61.61; H,6.73; N, 12.95.

Example 220

BOC-Trp-HLys (ω-N-Cbz)-Asp-Phe-NH₂

The compound is prepared by hydrogenating BOC-3,4-dehydro-L-homolysine (described in Example 219a) using 10% palladium-on-carbon in methanol under hydrogen atmosphere. The catalyst is removed by filtration through Celite and the solvent evaporated in vacuo. The resulting product is dissolved in chloroform, cooled to 0°C then treated with an equivalent of benzyl chloroformate along with concomitant addition of sodium hydroxide solution to afford BOC-HLys (ω-N-Cbz) upon standard work-up. The BOC-amino acid is extended to the final product via the procedures described in Example 219a and Example 219b.

Example 221

BOC-Trp-HLys (ω-N-[4-hydrgxycinnamoyl])-Asp- Phe-NH₂

The compound is prepared by reacting BOC-HLys (described in Example 220) with 4-hydroxycinnamic acid N-hydroxysuccinimide ester and N-methylmorpholine in DMF. After standard work-up, the resulting BOC-HLys (ω-N-[4-hydroxycinnamoyl]) is extended to the final tetrapeptide via the procedure described in Example 219a and Example 219b. Example 222

BOC-Trp-Lys (ε-N-r3-Dvridyl-3-acrylyl])-Asp- (NMe)Phe-NH₂

The product of Example 104 (0.153 g), the active ester of Example 38 (0.059 g), and N-methylmorpholine were allowed to react in the usual manner. The compound was purified as previously described in Example 216 to yield 80 mg of a white solid.

MS(FAB+) m/e 839 (M+H)⁺. ¹H NMR (DMSO-d₆, 30OMHz) δ 1.08-1.48

(m, 15H), 2.74-3.35 (m, 11H), 4.14-4.25 (m,2H), 4.62-4.68 (m, 1H), 4.84-5.02 (m,2H), 5.10-5.20 (m, 1H) , 6.68-6.83 (m,2H), 6.93-7.50

(m, 12H), 7.57-7.63 (m, 1H), 7.90-7.93 (m, 1H), 8.12-8.25 (m, 2H), 8.52-8.54 (m,1H), 8.71-8.73 (m, 1H) , 10.75-10.82 (m, 1H). Anal calcd for C₄₄H5₄N₈O₉ 1.5H₂O: C 61.03, H 6.63, N 12.

Example 223

Boc-Trp- (trans-5,6-dehydro)-L-2-Aminopimelic-(2-(4- hydroxyphenyl)-ethylamide)-Asp-Phe-NH₂ a . L-3-(2,2,2-Trichloroethoxycarbonyl)-5-oxo-4-oxazolidine propionic acid

To an ice cold suspension of L-glutamic acid (1.0 g, 6.8 mmol) in 40 ml dioxane-water (1/1 v-v) was added

diisopropylethylamme (DIEA) (3.7 mL, 21.1mmol) followed by 2,2,2-trichloroethylchloroformate (1.0 mL, 7.5 mmol). The mixture was subsequently allowed to warm to room temperature. After two hours the reaction mixture was partitioned between ethyl acetate and dilute aqueous HCl. The organic phase was dried (MgSO₄), filtered and concentrated in vacuo to yield a viscous oil which was sufficiently pure for use as isolated. The oil was combined with 2.2 ml 37% formalin solution and 100 mg p-toluenesulfonic acid in 50 ml toluene and the mixture heated to reflux with azeotropic removal of water. After approximately 3.5 hours the reaction mixture was allowed to cool then water washed , dried, filtered and concentrated in vacuo to give a pale yellow solid. The crude product was purified by flash chromatography on silica gel (ethyl acetate-hexane-acetic acid) to give 1.14 g (3.41 mmole) of a white solid. Anal. calc. for C₉H₁₀NO₆Cl₃: C, 32.29; H,3.02; N,4.18. Found: C,32.43; H,3.03; N, 4.08. MP 116-118.5°C. b. tert-Butyl (L-3-(2,2,2-trichloroethoxycarbonyl)- 5-oxo-4-oxazolidine)trans-2-pentenoate

The product from example 223a (1.12 g, 3.34 mmol) was

dissolved in anhydrous methylene chloride (25 mL) and treated with thionyl chloride (0.9 ml, 12.34 mmol). The flask was capped with a drierite filled drying tube and the mixture allowed to stir at room temperature for 6.5 hours. The volatiles were subsequently removed in vacuo. The residue was dissolved in a small volume of toluene and concentrated in vacuo a total of three times. The crude acid chloride so isolated was dissolved in anhydrous 1,2-dimethoxyethane (glyme) (45 mL) and, under nitrogen, cooled to - 78°C. To this was added via cannulae and nitrogen pressure a suspension of lithium tri-tert-butoxyaluminumhydride in 45 ml anhydrous glyme over a nine minute period. The mixture was allowed to stir seventeen minutes at -78°C before warming to room

temperature. After forty-five minutes the reaction mixture was quenched by cooling in an ice bath and adding (dropwise) dilute aqueous HCl. The mixture was poured into water and the product extracted with chloroform. The combined extracts were dried, filtered and concentrated in vacuo to yield the crude aldehyde as a pale yellow oil which was used without further purification. The aldehyde was dissolved in anhydrous methylene chloride (30 mL) and to this was added at room temperature a solution of tert-butoxycarbonyl-methylenetriphenylphosphorane ( 1.32 g, 3.51 mmol) prepared as in G.W.Kenner,et.al., Tetrahedron, 32 (2),275 (1976) in

30 mL anhydrous methylene chloride (16 mL) dropwise over one-half hour. The flask was capped with a drierite filled drying tube and allowed to stand at room temperature. After one hour, the mixture was concentrated in vacuo and the crude product purified by flash chromatography on silica gel (ethyl acetate- hexane) to give 0.68 g (1.63 mmol) of a white solid. Anal. calc. for C₁₅H₂₀NO₆Cl₃·H₂O : C,41.43; H,5.11; N,3.22. Found : C, 41.81; H,4.69; N,3.24. c . N-Boc-(trans-5,6-dehydro)-L-2-Aminopimelic(ε-2-(4-hydroxyphenyl) ethylamide)

The product from example 223b was treated with 1.5N HCl in glacial acetic acid (6 mL) at room temperature for one hour in a tightly capped flask. The reaction mixture was concentrated in vacuo and the residue dissolved in toluene and re-concentrated in vacuo a total of three times. The crude acid so isolated was coupled with tyramine (0.185 g, 1.35 mmol) via standard mixed anhydride methodology to give after flash chromatography on silica gel (ethyl acetate-hexane-acetic acid) 0.524 g (1.09 mmol) of the amide as a viscous oil. The amide was dissolved in tetrahydrofuran and to this solution at room temperature was added sodium

hydroxide (0.14 g, 3.6 mmol) in a total of 5 ml water. A further addition of isopropanol (2 mL) was necessary to give a homogeneous solution. After one hour the reaction mixture was concentrated in vacuo and the residue partitioned between ethylacetate and dilute aqueous HCl. The organic phase was washed once with brine then dried, filtered and concentrated in vacuo to give the crude acid 0.281 g (0.60 mmol) as an oil sufficiently pure for use as isolated. Dissolution in glacial acetic acid (10 mL) at room tempearture followed by treatment with a total of 1.37 g elemental zinc added in five increments over 3.5 h with sonication and protection from atmospheric moisture gave the N-deprotected alpha amino acid. The reaction mixture was vacuum filtered through celite and the filtrate concentrated in vacuo. The residue was suspended in methanol and concentrated in vacuo several times then finally suspended and vacuum filtered once again through celite. The filtrate was concentrated in vacuo to give a foam which was dissolved at room temperature in 10 mL of a dioxane-water (1/1 v-v) mixture containing DIEA (excess) and treated with Boc-carbonate (excess). The reaction mixture was allowed to stir overnight. The crude product was subsequently isolated by partition between ethyl acetate and dilute aqueous HCl. The organic phase was washed with water (1x), dried, filtered and concentrated in vacuo.

Purification by flash chromatography on silica gel (ethyl acetate-hexane-acetic acid then ethyl acetate-acetone-acetic acid) gave 0.061 g (0.155 mmol) of a clear oil. ¹H NMR (CDCl₃+DMSO-d₆, 300 MHz) δ 1.25 (s,9H), 1.6 (cm,1H), 1.79 (cm,1H), 2.06 (cm,2H), 2.53

(t,2H), 3.35 (m,2H), 4.03 (cm, 1H) , 5.3 (db, J=9Hz, 1H), 5.65

(db, J=15Hz,1H), 6.4-6.65 (m,3H), 6.82 (db, J=9Hz, 1H), 8.46 (bs, 1H).

_d. N-Boc-(trans-5,6-dehydro)-L-2-Aminopimelic(ε-2- (4-hydroxyphenyl)-ethylamide)-Asp-Phe-NH₂

The product from example 223c (0.061 g, 0.155 mmole) was coupled to Asp-Phe-NH₂ (added as the hydrochloride, 0.05 g, 0.155 mmol) via standard mixed anhydride methodology. The crude product was recrystallized from aqueous ethanol to give 0.049 g (0.075 mmol) of a white solid product. MS (FAB+) m/e 654 (M+H) + m/e 676 (M+Na)⁺. Anal. calc. for C₃₃H₄₃N₅O₉·2H₂O : C,57.45; H,6.88; N,10.15. Found: C, 57.03; H, 6.14; N,9.87. e. Boc-Trp-(trans-5,6-dehydro)-L-2-Aminopimelic (ε-2- (4-hydroxyphenyl)-ethylamide)-Asp-Phe-NH₂

The product from example 223d (0.046 g, 0.074 mmol) was treated with 1.5N HCl in glacial acetic acid (5 mL) at room temperature in a tightly capped flask. After one half hour the reaction mixture was frozen and lyophilized. The hydrochloride so isolated was combined with Boc-Trp-OSu (0.03 g, 0.074 mmol) in dimethylformamide (1 mL) at room temperature under nitrogen and treated with DIEA (0.028 mL, 0.163 mmol). The reaction mixture was allowed to stir overnight. Subsequent concentration in vacuo and recrystallization of the residue from aqueous ethanol gave 0.046 g

(0.055 mmol) of product as a white solid. MS (FAB+) m/e 840 (M+H) + m/e 862 (M+Na)⁺. ¹H NMR (DMSO-d₆, 300 MHz) δ 1.3 (s,9H), 1.63

(cm,2H), 2.07 (cm,2H), 2.4-3.4 (m, 10H), 4.1-4.45 (cm, 3H), 4,5

(m, 1H), 5.35 (db, J=15Hz, 1H), 6.58 (cm, 1H), 6.66 (db, J=7.5Hz, 2H),

6.8-7.4 (m, 15H), 7.59 (db, J=7.5Hz, 1H), 7.95 (m,3H), 8.3

(db, J=7.5Hz,1H), 9.17 (S,1H), 10.8 (s,1H), 12.45 (bs,1H). Anal. calc for C₄₄H₅₃N₇O₁₀ · 1 . 5 H₂O : C, 60 . 95; H, 6 . 52 ; N, 11 .31 . Found :

C, 60 .58 ; H, 6.12 ; N, 11 . 14 ; found: C 61 . 00, H 6.35, N 12 .82 .

Example 224

BOC-Trp-Lys(ε-N-[4-sulphatyl-cinnamoyl])-ASP-(NMe)Phe-NH₂

The tetrapeptide of Example 106 was allowed to react as described in Example 17. Purification under identical conditions yielded the product as a white solid. MS (FAB-) m/e 932 (M-H)⁺. ¹H NMR (DMSO-d₆, 300MHz) δ 1.04-1.43 (m, 15H), 2.02-2.12 (m, 1H), 2.34- 2.42 (m,1H), 2.73-2.94 (m, 5H), 3.03-3.32 (m, 4H), 4.16-4.30 (m,2H), 4.61-4.70 (m,1H), 4.84-4.92 (m, 1H) , 4.93-5.02 (m, 1H) , 5.09-5.17 (m,1H), 6.43-6.57 (m, 2H), 6.78-6.84 (m, 1H), 6.93-7.52 (m, 12H), 7.58-7.64 (m,1H), 7.80-7.93 (m, 1H), 8.00-8.09 (m, 2H), 8.27-8.31 (m, 1H) , 8 . 59-8 . 63 (m, 1H) . Anal calcd for C₄₅H₅₅N₇O₁₃S 2H₂O 0 . 9NH₃ : C 54 . 85 , H 6 . 31 , N 11 . 23 ; found : C 54 . 61 , H 6 . 08 , N 10 . 85 .

Example 225

BOC-Trp-Lys (ε-N- [2-methylphenylaminocarbonyl ] ) - Asp(β-OMe)-Phe-NH₂

The compound was prepared in a similar process to that described for Example 180 except that BOC-Asp (β-OMe) ester was used instead of BOC-Asp (β-OBn) ester. MS(FAB+) m/e 855 (M+H)⁺. NMR (DMSO-d₆, 300MHz) δ 1.10-1.46 (m, 15H), 1.91 (s,3H), 2.72-2.94 (m,7H), 3.00-3.36 (m, 5H), 3.454 (d,2H), 4.48-4.53 (m, 1H), 4.90-5.00 (m,2H), 5.38-5.43 (m, 1H), 6.48-6.53 (m, 1H), 8.81-7.01 (m,3H) 7.02-7.28 (m,9H), 7.29-7.34 (m,2H), 7.47 (s,1H), 7.54-7.63 (m,3H) 7.78-7.98 (m,2H), 8.28-8.32 (m, 1H). Anal calcd for C₄₅H₅₈N₈O₉ 0.5H₂O: C 62.42, H 6.83, N 12.66; found: C 62.26, H 6.80, N 12.90.

Example 226

Trp-Lys (ε-N-[2-methylphenylaminocarbonyl])-Asp- (NMe)Phe-NH₂ Hydrochloride

Z-Trp was coupled with Lys (ε-N-[2-methylphenylaminocarbonyl]Asp-Phe-NH₂ via the standard mixed anhydride coupling procedure using isobutylchloroformate. The product was dissolved in acetic acid and hydrogenated under a balloon of hydrogen gas in the presence of 10% palladium-on-carbon. After 3h, the catalyst was filtered and the acetic acid was evaporated. The residue was dissolved in HCl in acetic acid and lyopholyzed to a white powder. MS(FAB+) m/e 741 (M+H)⁺ . ¹H NMR (DMSO-d₆, 300MHz) δ 1.20-1.48

(m,6H), 2.72-3.27 (m, 14H), 4.03-4.10 (m, 1H), 4.20-4.38 (m, 1H), 4.62-4.93 (m,1H), 4.97-5.09 (m, 1H), 6.61-6.72 (m, 1H), 6.83-6.87 (m,1H), 6.93-7.28 (m, 1H), 7.47-7.52 (m, 1H), 7.70-7.83 (m,4H), 7.84-7.89 (m,1H), 7.98-8.03 (m, 2H) , 8.48-8.52 (m, 1H), 8.70-8.72 (m, 1H), 10.97-11.02 (m, 1H) . Anal calcd for C₃₉H₄₈N₈O₇ 2HCl 1.5H₂O: C 55.71, H 6.35, N 13.33; found: C 55.53, H 6.22, N 13.15.

Example 227

D-Trp-Lys (ε-N-[2-methylphenylaminocarbonyl])-Asp-(NMe)Phe-NH₂ Hydrochloride

The compound was prepared in an identical manner as described in Example 226 using Z-D-Trp. MS(FAB+) m/e 741 (M+H)⁺ . ¹H

NMR (DMSO-d₆, 300MHz) δ 1.00-1.38 (m, 6H), 2.25-2.41 (m, 1H), 2.72-3.28

(m,13H), 4.03-4.10 (m, 1H), 4.19-4.34 (m, 1H), 4.55-5.01 (m, 1H), 4.94-5.14 (m,1H), 6.58-6.68 (m, 1H), 6.84 (t, 1H), 6.95-7.50

(m, 16H), 7.62-7.84 (m, 2H), 8.04-8.12 (m, 1H), 8.40-8.43 (m, 1H), 8.68-8.84 (m, 1H) , 11.02 (m, 1H) . Anal calcd for C₃₉H₄₈N₈O₇ 2HCl NH₃ H₂O: C 52.93, H 6.62, N 14.25; found: C 52.97, H 6.29, N 14.17.

Example 228

BOC-Trp-Lys (ε-N-[2-methylphenylaminocarbonyl])-β-Asp- Phe-NH₂ a. Boc-β-Asp (α-benzγl)-Phe-NH₂

Commercially available Boc-Asp(α-benzyl)-OH (0.25 g, 0.77 mmol) was coupled to Phe-NH₂ (0.13 g, 0.77 mmol) via standard mixed anhydride methodology. The reaction mixture was partitioned between ethyl acetate and diluted with aqueous sodium bicarbonate. The organic phase was dried (MgSO₄), filtered and concentrated in vacuo to give 0.357 g (0.76 mmol) of crude product sufficiently pure for further use. MS (El) m/e 470 (M+H)⁺.

b . Nα-CBZ , Nε-Boc-Lys-β-Asp (α-benzvl ) -Phe-NH₂

The product from example 228a (0.357 g, 0.76 mmol) was treated with 1.5N HCl in glacial acetic (5 mL) at room temperature in a tightly capped flask. After fifty minutes the reaction mixture was frozen and lyophilized. The residue was dissolved in methanol and concentrated in vacuo (2x). The hydrochloride so isolated was subsequently coupled to N^α-CBZ,N^ε-Boc-Lys-OH (0.28 g, 0.75 mmol) via standard methodology (EDCI/HOBt). The reaction mixture was diluted with ethyl acetate and the organic phase washed (lx each) with dilute aqueous HCl, sodium bicarbonate and finally water. Upon drying, the product precipitated requiring concentration in vacuo to remove volatile organics followed by suspension of the residue in water to remove the inorganic/water soluble impurities. The mixture was vacuum filtered and the filter cake washed with water then dried in vacuo at room temperature to give 0.485 g (0.663 mmol) of a white solid. MS (FAB+) m/e 732 (M+H)⁺. c. Nα-CBZ,Nε-(2-methylphenylaminocarbonyl)-Lys-β-Asp(α-benzyl)-Phe-NH₂

The product from example 228b (0.479 g, 0.655 mmol) was treated with 1.5N HCl in glacial acetic acid (8 mL) at room temperature in a tightly capped flask. After one hour the reaction mixture was frozen and lyophilized. The hydrochloride was

subsequently treated at room temperature with o-tolyl isocyanate (0.080 ml, 0.655 mmol) in DMF (6 mL) containing DIEA (0.126 mL, 0.721 mmol). After approximately 2.5 the reaction was quenched by the dropwise addition of dilute aqueous HCl (100 mL) with vigorous stirring. The crude product 0.483 g (0.631 mmol), a white solid. was sufficiently pure for further use as isolated. MS (FAB)+ m/e 765 (M+H)⁺ m/e 787 (M+Na)⁺. d. Boc-Trp-Lys(ε-2-methylphenylaminocarbonyl)- β-Asp-Phe-NH₂

The product from example 228c (0.478 g, 0.63 mmol) was dissolved in glacial acetic acid (25 mL) and subjected to

hydrogenolysis (10% Pd/C, 4 atm. H₂) at room temperature. Filtration through celite followed by concentration in vacuo gave the crude product as a white solid which was used without further

purification. The material so isolated was combined with Boc-Trp-OSu (0.25 g, 0.63 mmol) in DMF (8 mL) containing DIEA (0.22 mL, 1.26 mmol) under nitrogen at room temperature. After 1.5, the reaction mixture was added dropwise to a rapidly stirring solution of dilute aqueous HCl. The resulting suspension was vacuum

filtered and the filter cake washed with water. The crude product was recrystallized from aqueous ethanol to give 0.267 g (0.323 mmol) of a white solid. MS (FAB+) m/e 827 (M+H) + m/e 849 (M+Na)+. ¹H NMR (DMSO-d₆,300 MHz) δ 1.1-1.7 (m, 6H), 1.3 (s,9H), 2.16 (s,3H),

2.36 (dd, J=15Hz,6Hz,1H), 2.55 (dd, J=15Hz, 6Hz, 1H), 2.7-3.15 (m, 6H), 4.22 (cm, 1H), 4.38 (cm,2H), 4.51 (cm, 1H), 6.55 (bs,1H), 6.8-7.45 (m, 16H), 7.6 (db, J=7.5Hz, 2H), 7.82 (db, J=7.5Hz, 1H), 7.9

(db,J=9Hz,1H), 8.2 (db, J=7.5Hz, 1H), 8.25 (db,J=7.5Hz,1H), 10.77 (s,1H), 12.64 (bs,1H). Anal. calc. for C₄₃H₅₄N₈O₉·0.5H₂O : C,61.77; H, 6.64; N,13.41. Found : C,61.43; H,6.52; N, 13.15.

Example 229

Ac-Trp-Lys (ε-N-[2-methylphenylaminocarbonγl])-Asp-Phe-NH₂

To a solution of BOC-Trp-Lys (Z)-Asp(Bn)-Phe-NH₂ (0.52 g) in acetic acid (5 mL) was added 1.4 N HCl in acetic acid (5 mL). The mixture was stirred for 2 h, the acetic acid removed in vacuo and the resulting residue taken up in THF (10 mL) and cooled to 0° C. Acetic anhydride (0.064 mL) and N-methylmorpholine (0.075 mL) were added, the reaction allowed to warm to ambient temperature and stirred overnight. The reaction was poured into water and

extracted three times with ethyl acetate which was washed with phosphoric acid solution, bicarbonate solution and brine followed by drying with sodium sulfate. After removal of solvent by evaporation, the residue was dissolved in acetic acid to which 100 mg of 10% palladium on carbon was added and stirred under a balloon of hydrogen gas for 4 h. The catalyst was filtered and the residue lyopholyzed from water. The resulting product was reacted with 2-methylphenyl isocyanate and N-methylmorpholine in DMF for 16 h. The reaction mixture was diluted with 25%

isopropanol in chloroform and washed with phosphoric acid. After drying over sodium sulfate, the solvents were evaporated and the residue purified on a preparative reverse phase C-18 column using acetonitrile and 0.05 M ammonium acetate solution (pH 4.5) as the eluants. MS (FAB+) m/e 769 (M+H)⁺. ¹H NMR (DMSO-d₆, 300MHz) δ 1.20- 1.55 (m,6H), 1.78 (s,3H), 2.16 (s,3H), 2.43-2.75 (m, 3H) , 2.80-3.1 (m,5H), 4.16-4.27 (m, 1H), 4.31-4.38 (m, 1H), 4.45-4.58 (m,2H), 6.50-6.53 (m,1H), 6.85 (t,1H), 6.95 (t,1H), 7.02-7.34 (m, 14H), 7.58-7.63 (m,1H), 7.78-7.86 (m, 1H), 8.07 (t, 1H) , 8.18 (d, 1H), 10.78 (s,1H). Anal calcd for C₄₀H₄₈N₈O₈ 4.5H₂O: C 56.86, H 6.20, N 13.26; found: C 57.19, H 5.95, N 12.77.

Example 230

BOC-Trp- (NMe ) Lys (ε-N- [2-methylphenylaminocarbonyl ] ) - Asp-Phe-NH₂ a. Boc-(N-Me)Lys(ε-N-[2-methγlphenylaminocarbonyl])-OH

A solution of 1.25 g, 3.1 mmol of Z-(N-Me)Lys(phthaloyl)-OH (Freidinger, R.M.; Hinkle, J.S.; Perlow, D.S.; Arison, B.H. J. Org. Chem. 1983,48,77-81) in 4 mL of MeOH was added to a

suspension of 100 mg of 10% Pd/C in 4 mL of MeOH, and the mixture was stirred under an atmosphere of H₂ for about 18 h. The resultant mixture was diluted with 8 mL of H₂O, treated with NEt₃ (457 μL, 3.3 mmol) and di-tert-butyl-dicarbonate (720 mg, 3.3 mmol), and stirred overnight. The mixture was diluted with aqueous HOAc and filtered. The filtrate was concentrated, and the remaining aqueous solution was extracted with EtOAc. The combined organic extracts were washed with brine, dried (Na₂SO₄) and evaporated to 280 mg of oily residue. A 239 mg (0.64 mmol) sample of the product was dissolved in absolute EtOH and treated with 34 μL (0.7 mmol) of hydrazine hydrate. The solution was heated under reflux for 1h, whereupon an additional 34 μL of hydrazine hydrate was added and heating under reflux was continued for 2 h. The mixture was concentrated under vacuum, diluted with aqueous HOAc, and filtered to remove precipitated phthalhydrazide. The filtrate was washed with EtOAc, and lyophilized to afford 190 mg of white powder, which was dried under reduced pressure at 50° C. A 185 mg (0.76 mmol) portion of the product was suspended in dry DMF and treated with NEt3 (115 μL, 0.83 mmol) and o-tolylisocyanate (104 μL, 0.83 mmol). The mixture was stirred for 1 h, then treated with additional NEt₃ (115 μL, 0.83 mmol) and o-tolylisocyanate (104 μL, 0.83 mmol) and stirred for an additional 0.25 h, whereupon the mixture was diluted with EtOAc and aq. NaHCO₃. The layers were mixed and separated, then the aqueous layer was acidified with aq. KHSO₄ and extracted with EtOAc. The organic layer which contained acidic product was dried over Na₂SO₄ and evaporated to 258 mg of crude product, which was

chromatographed over silica gel eluting with 10:9:1

hexane/EtOAc/HOAc to afford 125 mg of pure product. MS (CI/NH₃) m/e 394 (M+H)⁺, 411. 1H NMR(CDCl₃) δ 1.32 (m, 2H), 1.45 (s, 9H), 1.52 (m,1H), 1.75 (m,1H), 1.97 (m, 1H) , 2.29 (s,3H), 2.80 (br s,3H), 3.22 (m,2H), 4.43 (m,0.4H), 4.70 (m, 1.6H), 7.0 (s,1H), 7.07-7.35 (m, 4H). b. H-(N-Me)Lys(ε-N-[2-methylphenylaminocarbonyl])-Asp- Phe-NH₂ CF₃CO₂H

The product of Example 230a (53 mg, 0.135 mmol) was coupled to the TFA salt of H-Asp(Bzl)-Phe-NH₂ (98 mg, 0.2 mmol) by the usual mixed isobutylcarbonic anhydride procedure, to afford the crude product in quantitative yield. Treatment with 1:1 TFA/CH₂CI₂ at room temperature for 1 h, followed by evaporation of volatile components and precipitation of the product with anhydrous Et₂O provided 85 mg (85%) of the title compound. MS (FAB+) m/e 659 (M+H)⁺, 681 (M+Na) ⁺. ¹H NMR (DMSO-d₆) δ1.25 (m, 2H), 1.40 (m,2H),

1.68 (m,2H), 2.18 (s,3H), 2.38 (s, 3H), 2.63 (dd, J= 9 and 16.5 Hz, 1H), 2.80 (dd, J= 9 and 14 Hz, 1H), 2.9 (dd, J= 4.5 and 16.5 Hz, 1H), 3.01 (m,2H), 3.61 (m, 1H), 4.42 (m, 1H), 4.78 (m, 1H), 5.1 (s,2H), 6.52 (m,1H), 6.86 (t, J= 15 Hz,1H), 7.05-7.30 (m, 8H), 7.38 (m, 5H), 7.45 (s,1H), 7.60 (s,1H), 7.80 (d, J= 7.5 Hz,1H), 8.2

(d, J= 8 Hz, 1H), 8.88 (d,J=8 Hz,1H).

c. Boc-Trp-(N-Me)Lys(ε-N-methylphenylaminocarbonyl])- Asp-Phe-NH₂

The product of Example 230b (43 mg, 0.057 mmol) was coupled with the symmetrical anhydride prepared from Boc-Trp-OH (69 mg, 0.228 mmol) in CH₂CI₂/DMF in the presence of diisopropylethylamine (10 μL, 0.06 mmol). Extractive work-up and chromatography over silica gel eluting with 4:96 MeOH/CHCl₃ afforded 40 mg of protected tetrapeptide. Catalytic hydrogenolysis (H₂, 10% Pd-C, MeOH) of 36 mg of the above product, followed by recrystallization of the crude product from MeOH/H₂O afforded 22 mg of the title compound. MS (FAB+) m/e 863, 841, 741. ¹H NMR (DMSO-d₆, 100° C) δ 1.15-1.40 (m, 11H, includes δ 1.30, s, 9H) , 1.45 (br s,2H), 1.60 (br s,1H), 1.8 (br s,1H), 2.18 (s,3H), 2.5 (1H, obscured), 2.65 (m, 1H) , 2.8-3.5 (9H, obscured), 4.45 (q,J= 3 Hz, 1H) , 4.55 (q,J=4.5 Hz,1H), 4.66 (q,4 Hz,1H), 4.90 (br s,1H), 6.22 (br s,1H), 6.38 (br s,1H), 6.87 (t, J=4.5 Hz, 1H), 6.93 (t,J= 4.5 Hz), 7.05-7.15 (m, 4H) , 7.2

(m,4H), 7.33 (d, J= 5 Hz,1H), 7.39 (s,1H), 7.55 (m,2H), 7.71 (d,J= 4.5 Hz,1H). ¹H NMR (DMSO-d₆, 65°C) reveals N-CH₃ at δ 2.87. Anal.

Calcd for C₄₄H₅₆N₈O₉·2H₂O: C, 60.26; H, 6.89; N, 12.77; found: C, 60.31, H, 6.40; N, 12.54.

Example 231

BOC-Trp-(NMe)Lys(ε-N-[2-methylphenylaminocarbonyl])- Asp-(NMe)Phe-NH₂ a. H-(N-Me)Lys-Asp(Bzl)-(N-Me)Phe-NH₂.CF₃CO₂H

The product of Example 230a (67 mg, 0.17 mmol) was coupled to the HCl salt of H-Asp(Bzl)-(N-Me)Phe-NH₂ (107 mg, 0.26 mmol) by the usual mixed isobutylcarbonic anhydride procedure, to afford the crude product in quantitative yield. Treatment with 1:1 TFA/CH₂CI₂ at room temperature for 1 h, followed by evaporation of volatile components and precipitation of the product with anhydrous Et₂O provided 96 mg (72%) of the title compound. MS (FAB⁺) m/e 659 (M+H⁾⁺, 681 (M+NH₄)⁺. ¹H NMR(DMSO-d₆) (two conformers) δ 1.2 (m, 2H), 1.4 (m, 2H), 1.57 (m, 1H) , 1.7 (m, 1H) , 2.15 and 2.17 (two s,3H), 2.4 (two br s,3H), 2.65 (dd, J = 8 and 16 Hz,1H), 2.8

(s,1H), 2.9 (m,1H), 2.95 (s,2H), 3.04 (m, 2H) , 3.15-3.3 (m, 2H, partially obscured), 3.62 (m, 1H), 4.22 (m, 1H) , 4.33 (m, 1H) , 5.02-5.11 (m,2H), 5.13 (m, 1H) , 6.52 (m, 1H) , 6.87 (t,J= 6.5 Hz, 1H), 7.1 (m, 3H), 7.2 (m, 5H) , 7.4 (m, 5H) , 7.6 (m, 2H), 7.8 (m, 1H) , 8.8 (br s), 9.0 (d,J= 9 Hz,0.5H), 9.19 (d, J= 6 Hz, 0.5 H). b. BOC-Trp- (NMe) Lys (ε-N- [2-methylphenylaminpcarbonyl] ) - Asp-(NMe)Phe-NH₂

The product of Example 231a (50 mg, 0.065 mmol) was converted to the title compound by a procedure analogous to that of Example 231a. The crude product was purified by chromatography over silica gel eluting with 62:3:2:1 EtOAc/pyridine/H₂O/HOAc to afford the product in a 50% overall yield. MS (FAB⁺) m/e 855

(M+H)⁺,755, 562. ¹H NMR (DMSO-d₆, 148°C) δ 1.15-1.45 (m, 13H, includes d 1.32 (s, 9H)), 1.75 (m,2H), 2.05 (s,0.5H), 2.16 (s,2.5H), 2.35 (m, 1H), 2.6-2.85 obscured, 2.9 (s,3H), 2.95 (dd,J= 5 and 9

Hz,1H), 3.05 (m,1H), 3.10 (dd,J= 3 and 9 Hz,1H), 3.28 (dd,J= 3 and 9 Hz, 1H) , 4.7 (m, 2H), 4.95 (m, 1H), 5.05 (m, 1H), 6.03 (br m,1H), 6.10 (br m,1H), 6.61 (m, 2H) , 6.87 (m, 1H) , 6.96 (t,J= 4 Hz,1H), 7.05 (t,J= 4.5 Hz,2H), 7.08 (br s,2H), 7.15-7.30 (m,7H), 7.32 (d,J= 5 Hz,1H), 7.64 (d,J= 5 Hz,1H), 10.35 (br s,1H). Anal. Calcd for C₄5H₅₈N₈O₉Η₂O: C, 61.91, H, 6.93, N, 12.84; found: C, 61.96, H, 6.81, N, 12.62.

Example 232

BOC-Trp-Lys (ε-N- [2-methylphenylaminocarbonyl ]) - (NMe)Asp-Phe-NH₂ a. Fmoc-(N-Me)Asp(Bzl)-OH

According to the general procedure described in Freidinger, R.M.; Hinkle, J.S.; Perlow, D.S.; Arison, B.H. J. Org. Chem.

1983,48,77-81, 15 g of Fmoc-Asp(Bzl)-OH was converted to the title compound (obtained as a solid after trituration with Et₂O/hexane) in 78% yield. MS (FAB⁺) m/e 460 (M+H)⁺, 482 (M+Na)⁺. ¹H NMR (CDCI₃) (two conformers) δ 2.35 (dd,J= 7.5 and 17 Hz, 0.5 H), 2.72

(dd,J= 6 and 16 Hz, 0.5 H) , 2.6 (s,1.5 H) , 2.95 (1H, obscured), 3.0 (s,1.5 H), 3.18 (dd, J= 6 and 17 Hz, 0.5 H) , 4.15 (m, 0.5 H), 4.25 (t,J= 7.5 Hz,0.5H), 4.42 (m, 1.5 H), 4.5 (dd,J= 5 and 10.5 Hz, 0.5 H), 4.62 (m, 0.5 H), 4.8 (dd,J= 6 and 8 Hz, 0.5 H) , 5.13

(m,2H), 7.3 (m, 9H), 7.52 (m, 2H) , 7.68 (d,J= 7.5 Hz,0.5 H), 7.75

(d, J= 7.5 Hz, 1.5 H). b. Fmoc-(N-Me)Asp(Bzl)-Phe-NH₂

The product of Example 232a (4.45g, 9.69 mmol) was coupled to Phe-NH₂ (1.6g, 9.69 mmol) using BOP reagent (5g, 11 mmol) and N-methylmorpholine (1.1 mL, 10 mmol) in DMF, with stirring at 0°C followed by warming to room temperature and stirring overnight. The mixture was concentrated, diluted with EtOAc, and the solution was washed with aqueous citric acid, aqueous NaHCO₃, and brine, dried (MgSO₄) and evaporated to 2.88 g (49%) of product. MS (FAB⁺) m/e 606 (M+H⁺) , 628 (M+Na⁺). ¹H NMR (CDCI₃) (two

conformers) δ 2.34 (m, 1H) , 2.46-2.70 (m, 4H, includes 2.59,s), 2.99 (m,1H), 3.1-3.3 (m,2H), 4.22 (m,2H), 4.48 (m, 1H) , 4.70 (m, 1H) , 5.0-5.15 (m,2H), 5.32 (major) and 5.41 (br m's,1H), 6.05 (major) and 6 . 25 (br m ' s , 1H) , 6 . 51 (d, J=7 . 5 Hz , 1H) , 6 . 9-7 . 5 (m, 14H) , 7 . 56 (d, J=7 . 5 Hz , 1H) , 7 . 8 (d, J=7 . 5 Hz , 1H) . c . Boc-Trp-Lys(ε-N-[2-methylphenylaminocarbonyl1)-(N- Me)Asp-Phe-NH₂

The product of Example 232b (510 mg, 0.84 mmol) was treated with 50% diethylamine/ CH₃CN for 40 min., then the volatile components were evaporated under reduced pressure. Additional CH₃CN was added and evaporated to afford 438 mg of crude product. A 225 mg portion of the crude product in 2 ml of DMF at 0°C was treated with the symmetrical anhydride prepared from 407 mg (1.07 mmol) of Boc-Lys(ε-N-[2-methylphenylaminocarbonyl])-OH and EDCI (103 mg, 0.54 mmol, in CH₂CI₂ at 0°C). The solution was allowed to warm to room temperature and stir for about 18 hr. After extractive work-up, the crude product was chromatographed over silica gel, eluting with 5% MeOH/CHCl₃ to afford 160 mg (50%) of pure protected tripeptide, MS (FAB⁺) m/e 745, 767. Elaboration to the crude deprotected tetrapeptide was carried out in the usual fashion (Boc deprotection, active ester coupling, benzyl ester hydrogenolysis), and the final product was purified by

chromatography over silica gel, eluting with 56:3:2:1

EtOAc/pyridine/H₂O/HOAc. Pure fractions were combined,

concentrated, diluted with H₂O and lyophilized to afford 42 mg of pure product. MS (FAB⁺) m/e 841 (M+H⁺) , 741, 577. ¹H NMR(DMSO-d₆ (two conformers) δ 1.05-1.9 (m, 15H, includes 1.30, s and 1.31, s,total 9H), 2.10-2.25 (m, 5H, includes 2.15,s, and 2.20, s), 2.58 (m, 1.5 H) , 2.7 (dd,J= 6 and 9 Hz,0.5H), 2.75-3.2 (m,3H), 3.41 (0.5 H, obscured), 4.21 (m,0.5 H) , 4.32 (m, 1H) , 4.40 (m,0.5 H), 4.59 (br m,1H), 4.9 (m,0.5H), 5.08 (m,0.5H), 5.22 (m,0.5H), 6.67 (m,0.5H), 6.82 (m, 1H) , 6.87 (m,0.5H), 6.95 (m, 1H) , 7.05 (m, 4H) , 7.11-7.28 (m,5.5H), 7.30 (d,J=5 Hz, 1H), 7.45 (s,0.5H), 7.50 (s,0.5H), 7.57 (t,J=4 Hz,1H), 7.18 (d, J=5 Hz,0.5H), 7.63 (br s,0.5H), 7.81 (d, J=5 Hz,0.5H), 8.02 (d,J= 5 Hz,0.5H), 8.10 (br m,0.5H), 8.26 (br m,0.5H), 8.35 (m, 1H) , 8.45 (br m,0.5H), 8.59 (br m,0.5H), 8.81 (br m,0.5H), 10.8 (s,0.5H), 10.97 (br s,0.5H).

Anal. Calcd. for C₄₄H₅₆N₈O₉-O.8HOAc·1.2H₂O: C, 60.14, H, 6.81, N, 12.31; found C, 60.06, H, 6.49, N, 12.37.

Example 233

BOC-Trp-Lys(ε-N-[2-methylphenylaminocarbonyl])- (NMe)Asp-(NMe)Phe-NH₂ a. Fmoc-(N-Me)Asp(Bzl)-(N-Me)Phe-NHo

The product of Example 232a (540 mg, 1.18 mmol), CF₃COOH·H- (N-Me)Phe-NH₂ (344 mg, 1.18 mmol) and NEt₃ (538 μL, 3.89 mmol) were combined in CH₂CI₂ at 0°C then BOP-Cl was added. The mixture was allowed to warm to room temperature and stirred overnight. The solution was diluted with EtOAc and subjected to standard acid-base extractive work-up, followed by chromatography of the crude product over silica gel, eluting with 2:1 hexane/acetone to afford 360 mg (49%) of the title compound. MS (FAB⁺) m/e 642 (M+Na⁺) . ¹H NMR(CDCl₃) (multiple conformers) δ 1.9-3.0 (m, 11H, methyl singlets at 1.92, 2.09, 2.21, and 2.82), 3.12 (m, 1H) , 3.40 (m,1H), 4.19 (m, 1H), 4.35-4.45 (m, 2H) , 4.6-4.8 (m, 1H) , 4.98-5.18 (m, 2H) , 5.32-5.68 (m,2H), 6.08-6.29 (m, 1H) , 6.98-7.62 (m, 15H) , 7.65-7.87 (m, 3H) . b. Boc-Trp-Lys(ε-N-[2-methylphenylaminocarbonyl])-(N- Me) ASP-(N-Me)Phe-NH₂

The product of Example 233a was extended to the title

compound by a procedure analogous to that described in Example 232c. MS(FAB⁺) m/e 577 (M+H) ⁺. ¹H NMR(DMSO-d₆) δ 1.15 (m, 1H) , 1.20-1.35 (m,10 H, includes 1.30 (s, 9H) ) 1.35-1.52 (m, 4H) , 2.0 (dd,J= 2 and 10 Hz, 1H), 2.06 (s,3H), 2.17 (m, 4H, includes s,3H), 2.75 (m, 1H and s,3 H), 2.9 (m, 2H), 3.04 (dd J= 3 and 9 Hz,1H), 3.1 (m, 1H) , 3.21 (dd,J=3 and 9 Hz,1H), 4.2 (m, 1H), 4.42 (q, J=4Hz, 1H), 5.33 (dd,J= 2 and 7 Hz,1H), 5.51 (dd,J=3 and 9 Hz,1H), 6.58 (t,J=3 Hz,1H), 6.73 (d, J=5 Hz,1H), 6.85 (t,J=4 Hz, 1H), 6.95 (t,J=5

Hz,1H), 7.08 (m, 6H), 7.15-7.4 (m, 9H) , 7.59 (m, 2H), 7.83 (d, J=5 Hz,1H), 7.95 (d,J=5 Hz, 1H), 10.8 (s,1H). Anal. Calcd. for

C₄₅H₅₈N₈O₉.1.2 HOAc: C, 61.41, H, 6.82, N, 12.09; found: C, 61.36, H, 6.64, N, 12.17.

Example 234

BOC-Trp-Lys(ε-N-[2-methylphenylaminocarbonyl])- Asp-Phe-NHMe

To a solution of 50mg (0.17 mmol) of the TFA salt of H-Phe-NHMe (Suzuki, K. et al. Chem. Pharm. Bull. 36 (12), 4834, 1988), 132 mg (0.17 mmol) of Boc-Trp-Lys(ε-N-[2-methylphenylaminocarbonyl])-Asp (Bzl)-OH and 89 μL (0.51 mmol) of diisopropylethylamine in 3 mL of CH₂CI₂ at 0°C was added BOP-Cl (43 mg, 0.17 mmol). After stirring for 4h at 0°C, stirring was continued as the mixture was allowed to warm to room temperature. After standard acid-base extractive work-up, the crude product was chromatographed over silica gel eluting with 4% MeOH/CHCl₃ to afford 56 mg of protecte tetrapeptide. A 52 mg sample was subjected to catalytic transfer hydrogenolysis with HCOONH₄ in MeOH in the presence of 10% Pd-C. The mixture was filtered and evaporated, then purified by

chromatography over a C₁₈ reverse-phase column, eluting with

CH₃CN/50 mM NH₄OAC, pH 4.5. Pure fractions were combined and lyophilized twice to afford 17 mg of pure product. MS(FAB⁺) m/e 863 (M+Na⁺), 741 (M+H-Boc)⁺. ¹H NMR (DMSO-d₆) 8 1.1-1.7 (m, 15H), 2.17 (s,3H), 2.55 (d, J=4 Hz, 3H), 2.6-3.15 (m, 8H) , 4.25 (m,2H),

4.35 (m, 1H), 4.49 (m, 1H) , 6.61 (br s, 1H), 6.84 (m,2H), 6.95

(m,1H), 7.01-7.13 (m,4H), 7.15-7.28 (m, 6H) , 7.31 (d, J=9 Hz, 1H), 7.6 (d, J=9 Hz,1H), 7.82 (m, 2H), 7.96 (m,2H), 8.23 (m, 1H) , 10.81

(s,1H). Anal. Calcd for C₄₄H₅₆N₈O₉.1.5 H₂O: C, 60.89, H, 6.85, N, 12.91; found: C, 60.71, H, 6.63, N, 12.61.

Example 235

BOC-Trp-Lys(ε-N-[2-methylphenylaminocarbonyl])-Asp- Phe-NMe₂ a. HCl Asp(Bzl)-Phe-NMe₂

A solution of Boc-Asp (Bzl)-OH (158 mg, 0.49 mmol), the TFA salt of Phe-NMe₂ (Suzuki, K. et al. Chem. Pharm. Bull.

36(12), 4834,1988) (150 mg, 0.49 mmol), N-methylmorpholine (55 μL ,0.50 mmol) and HOBt.H₂O (75 mg, 0.49 mmol) in 3 mL of CH₂CI₂ at 0°C was treated with EDCI (94 mg, 0.49 mmol). The solution was allowed to warm to room temperature and stirred overnight. The solution was diluted with EtOAc and subjected to standard acidbase extractive work-up. The crude product was treated with 1:1 CF₃COOH/CH₂CI₂ for 1h, then the volatile components were

evaporated. A solution of HCl in dioxane was added to convert the product to the HCl salt which solidified upon addition of

anhydrous Et₂O and was collected by filtration to afford 185 mg of white solid. MS (CI) m/e 398. ¹H NMR(DMSO-d₆) δ 2.75 (s,3H),

2.78-2.9 (m, 5H, includes 2.81,s,3H), 2.97 (m, 2H) , 4.12 (m, H), 4.89 (m,1H), 5.15 (m,1H), 7.25 (m, 5H), 7.4 (m, 5H), 8.22 (br m,3H), 8.88 (d J=8 Hz,1H).

b. Boc-Trp-Lys (ε-N-[methylphenylaminocarbonyl])-Asp- Phe-NMe₂

A solution of HCl Asp (Bzl)-Phe-NMe₂ (90 mg, 0.21 mmol), Boc-Trp-Lys(ε-N-[methylphenylaminocarbonyl])-OH (117.3 mg, 0.21 mmol), HOBt.H₂O (32 mg, 0.21 mmol) and diisopropylethylamine (37 μL, 0.21 mmol) in 5 mL of 1:1 THF/CH₂CI₂ at -20°C was treated with EDCI (40 mg, 0.21 mmol). After keeping at -20°C for 2 days, the solution was allowed to warm to room temperature. The solvent was

evaporated and the mixture was subjected to standard acid-base extractive work-up to afford 197 mg of the crude protected

tetrapeptide. Hydrogenolysis of a 155 mg sample over 10% Pd/C was followed by chromatography over silica gel eluting with 56:3:2:1 EtOAc/pyridine/H₂O/HOAc. Pure fractions were combined,

concentrated, diluted with H₂O and lyophilized twice to afford 65 mg of the title compound. MS (FAB⁺) m/e 855 (M+H)⁺, 877 (M+Na)⁺, 893 (M+K)⁺. ¹H NMR (DMSO-d₆) δ 1.12 (m, 1H) , 1.30 (s,9H), 1.42

(m,3H), 1.54 (m, 1H) , 1.68 (m, 1H) , 2.15 (s,3H), 2.48

( 1H, obscured), 2.62 (dd,J = 3 and 9 Hz,1H), 2.8 (dd, J = 4 and 8 Hz,1H), 2.92 (dd, J = 4 and 9 Hz, 1H) , 3.02-3.18 (m,3H), 3.3

(1H, obscured), 4.23 (m, 1H) , 4.3 (m, 1H) , 4.54 (q, J=3Hz, 1H), 4.83 (q,J=4 Hz,1H), 6.59 (br s, 1H) , 6.79 (d, 5 Hz, 1H) , 6.85 (t,J=4.5 Hz,1H), 7.02-7.13 (m, 5H) , 7.15-7.21 (m, 4H), 7.23 (m, H) , 7.31 (d,J=5 Hz,1H), 7.59 (d,J=5 Hz,1H), 7.65 (br s,1H), 7.80 (m, 1H), 7.95 (m, 2H), 8.22 (d, J=4.5 Hz,1H), 10.78 (s, 1H). Anal. Calcd for C₄₅H₅₈N₈O₉-1.5 H₂O : C, 61.28, H, 6.97, N, 12.70; found: C,

61.66, H, 6.86, N, 12.42.

Examole 236

BOC-Trp-Lys (ε-N-[2-methylphenylaminocarbonyll) [ΨCH₂NH] - Asp-Phe-NH₂

The title compound was prepared in a similar manner to that described in Example 190 using 2-methylphenyl isocyanate in place of 4-hydroxycinnamic acid N-hydroxysuccinimide ester. MS (FAB+) m/e 813 (M+H)⁺. ¹H NMR (DMSO-d₆, 50 OMHz) δ 1.04-1.42 (m, 6H) , 1.23

(br s,9H), 1.98-2.00 (m, 1H) , 2.10 (s,3H), 2.19-2.29 (m, 2H) , 2.39 (dd, 1H), 2.73 (dd, 1H) , 2.83-2.91 (m, 1H) , 2.95-3.12 (m, 4H) , 3.38 (t,1H), 3.66 (m,1H), 4.10-4.17 (m, 1H) , 4.45 (dd, 1H) , 6.89 (t,1H), 6.95 (t,1H), 7.02-7.22 (m, 11H) , 7.32 (d, 1H) , 7.56 (br t,2E) . Anal calcd for C₄3H₅₆N₈O₈ C₂H₄O₂ 1.5H₂O: C 60.05, H 7.06, N 12.45;

found: C 60.17, H 6.65, N 12.75.

Example 237

2-Fluoro-3-(indol-3-yl)-propionyl-Lys(ε-N-[2-methylphenylaminocarbony ])-Asp-PheNH₂ a. 2-Fluoro-3-(indol-3-yl)-2-propenoic acid ethyl

ester

To a solution of triethyl-α-fluoro-phosphonoacetate (4.6 g,

19 mmol), LiCl (860 mg, 20 mmol) and DBU (3.2 mL, 23 mmol) was added indole-3-carboxaldehyde (2.9 g, 20 mmol). The reaction was left at ambient temperature overnight then added to 10% citric acid and extracted with CH₂CI₂. After drying over Na₂SO₄, the solution was filtered and solvent was removed in vacuo. The crude product was chromatographed (ethyl acetate/hexane) then

recrystallized (diethyl ether/hexane) to yield 3.1g (66%) of a white solid. MS(CI) 251 (M+NH₄)⁺. ¹H NMR(CDCl₃, 300MHz) δ 1.40

(2t,3H), 4.37 (q,2H), 7.20-7.32 (m, 4H) , 7.38-7.45 (m, 1H), 7.78-83 (m, 1H) , 8.52 (br d, 1H) . b. 2-Fluoro-3-(indol-3-yl)-propionic acid

The product from example 237a (500 mg, 2.14 mmol) and 10% Pd/C (50 mg) in methanol (20 mL) was hydrogenated at ambient temperature overnight. The catalyst was filtered and the solvent was removed in vacuo. The residue was redissolved in methanol (4 mL) then cooled to 0°C. 1N NaOH (4 mL) was added and the reaction stirred at ambient temperature for 10 h, acidified to pH 2-3 and extracted with ethyl acetate (4x). After drying over Na₂SO₄, the solution was filtered and the solvent was removed in vacuo. The residue was chromatographed (ethyl acetate/hexane with 2% HOAc) to yield 160 mg (36%) of a product. MS(CI) 225 (M+NH₄)⁺. ¹R

NMR (CD₃OD, 300MHz) δ 3.20-3.48 (m,2H), 5.17 (ddd, 1H), 7.01 (br t,1H), 7.08 (br t,1H), 7.13 (br s,1H), 7.32 (br d, 1H), 7.54 (br d,1H). c. 2-Fluoro-3-(indol-3-yl)-propionic acid

2 , 4, 5-trichlo rophenyl ester

To a solution of example 237b (104 g, 0.5 mmol) in 2 mL anhydrous CH₂CI₂ was added 2, 4, 5-trichlorophenol (150 mg, 0.75 mmol), HOBt (81 mg, 0.6 mmol) and EDCI (120 mg, 0.6 mmol). The solution was stirred at room temperature for 24 h then washed with 10% citric acid (1x), H₂O (1x) and brine. After drying over

Na₂SO₄, the solution was filtered and the solvent evaporated. The residue was chromatographed (ethyl acetate/hexane) to yield 125 mg of a white solid. MS(CI) 403, 405, 409 (M+NH₄)⁺. ¹H

NMR (CDCI₃, 300MHz) δ 3.56 (ddd, 2H), 5.45 (ddd, 1H), 7.13-7.30 (b m, 5H), 7.42 (br d, 1H), 7.64 (br d, 1H), 8.15 (br s, 1H).

d. 2-Fluoro-3-(indol-3-yl)-propionyl-Lys(g-N-[2-methylphenylaminocarbonyl])-Asp-PheNH₂

To a solution of the TFA salt of Lys (ε-N [2-methyIphenylaminocarbonyl])-Asp-PheNH₂ (210 mg, 0.32 mmol) in DMF (2 mL) at 4°C was added diisopropylethylamine (0.12 mL, 0.69 mmol) and the active ester of example 237c (124 mg, 0.32 mmol). The reaction mixture was stirred at ambient temperature for 10 h, then was poured into a cold, rapidly stirring solution of 10% citric acid causing a white precippate which was collected by filtration. The crude product was suspended in hot ethyl acetate for 30 min with vigorous stirring, cooled to room temperature and the solid was collected by filtration to yield 170 mg (73%) of a white solid. HPLC analysis (Cis-utrasphere ODS with CH₃CN/50 mM NH₄OAc buffer as eluent) showed a diastereomeric ratio of 1/1 at the carbon that bears the fluorine atom. MS(FAB+) m/e 730 (M+H)⁺. ¹H NMR (DMSO-d₆/D₂O, 300MHz) δ 1.21-1.62 (m, 6H) , 2.15(2s,3H), 2.30- 3.32 (m,8H), 4.26(m,1H), 4.37 (m, 1H) , 4.51(m,1H), 5.18(2m,1H),

6.87 (br t, 1H) , 6.97 (br t, 1H), 7.06-7.30 (m, 9H) , 7.34 (br d, 1H), 7.55 (br t, 1H), 7.84(2d,1H). Anal calc for C₃₈H₄₄FN₇O₇ · 0.5H₂O: C

61.78, H 6.14, N 13.27; found: C 61.48, H 6.09, N 12.91.

Example 238

2-Cyano-3-(indol-3-yl)-propionyl-Lys(ε-N-[2-methylphenylaminocarbonyll)-Asp-PheNH₂

The corresponding 2-cyano derivative is prepared in a similar fashion described in Example 237 from the known 2-cyano-3-(1H-indol-3-yl)-2-propenoic acid ethyl ester. [Chem. Pharm. Bull.

1980, 28(6), p.1711-1721., and Chem. Ber.. 1977, 110(1) p.86-95].

Example 239

Boc-Trp-Lys (ε-N-[2-methylphenylaminocarbonyl])-Asp-Phe-OMe

N-Methylmorpholine (50 mL) followed by iso-butylchloroformate (65 mL) were added to a solution of the dipeptide Boc-Trp-Lys (ε-N- [2-methylphenylaminocarbonyl])-OH (265 mg) in 5 mL of THF stirring at -15°C. After stirring for 20-25 minutes additional NMM (50 mL) was added to the reaction mixture followed by Asp (OBn)-Phe-OMe hydrochloride (228 mg, prepared in a similar fashion to the hydrochloride of Example 2) as a neat solid. The reaction vessel was maintained at low temperature (-15°C to -10°C) for 4-5 hours then allowed to warm to ambient temperature overnight. The mixture was poured into ethyl acetate and washed successively with water, dilute aqueous hydrochloric acid, saturated aqueous sodium

bicarbonate, and water. After concentration of the solution, some product (185 mg of benzyl ester tetrapeptide) was isolated by trituration with ethanol. Chromatography of the concentrated filtrate provided additional pure benzyl ester tetrapeptide.

The benzyl ester (265 mg) in 3.5 mL of DMF was added

via cannula to a suspension of 10% palladium on carbon in 4 mL of DMF at ambient temperature under nitrogen atmosphere

that had been treated with 100 mL of cyclohexadiene. When

the addition was complete, an additional 400 mL of

cyclohexadiene was added and stirring continued overnight

at which time tic indicated consumption of starting benzyl

ester. The reaction mixture was filtered through celite

with ethanol as a co-solvent and the filtrate was

concentrated in vacuo. Chromatography of the residue using a pyridine:acetic acid:water:ethyl acetate solvent system

provided fractions with pure product. These fractions were concentrated in vacuo, diluted with water and lyophilized

to provide desired tetrapeptide. MS (FAB) m/e 864(m+Na)⁺, 842(m+H)⁺, 780, 742. ¹H NMR(300MHz,DMSO-d₆, 135°C) δ 10.4

(bs,1H), 7.85-7.90 (bs, 1H), 7.68 (bd, J=7Hz, 1H) , 7.56

(bd, J=7Hz, 1H), 7.45-7.55 (m, 2H), 7.36 (bd, J=8Hz, 1H), 7.15-7.30 (m, 4H), 7.0-7.15 (m, 4H) , 6.97 (dt, J=1, 7Hz, 2H), 6.88 (dt,J=l,7Hz,1H), 6.26 (bs, 1H) , 6.14 (bd, J=8Hz, 1H), 4.58 (m,2H), 4.30 (m,2H), 3.59 (s,3H), 2.95-3.25 (m, 7H) , 2.90 (bs,H₂O), 2.62 (dd, J=7,15Hz,1H), 2.5-2.58 (m, 1H) , 2.2

(s,3H), 1.4-1.8 (m, 6H), 1.35 (s, 9H). Anal calcd for

C₄₄H₅₅N₇O₁₀ 1.1 HOAc: C 61.11, H 6.59, N 10.80; found: C 61.11, H 6.50, N 10.89.

The compounds of formula I are CCK agonists which are useful in the treatment and prevention of CCK-related disorders of the gastrointestinal, central nervous, and appetite and insulin regulatory systems of animals and humans. As CCK agonists, they are useful in the treatment and prevention of neuroleptic disorders, tardive

dyskinesia, disorders of memory and cognition, Parkinson's disease, Huntington's chorea, psychosis, including

schizophrenia, Gilles de la Tourette syndrome, diabetes, disorders of appetite regulatory systems, obesity, bulimia, the treatment of pain and the treatment of substance abuse.

The ability of the compounds of the invention to interact with CCK receptors and to act as CCK agonists can be demonstrated in vitro using the following protocols.

CCK₈ [Asp-Tyr (SO₃H) -Met-Gly-Trp-Met-Asp-Phe-NH₂], bestatin and phosphoramidon were

purchased from Peptide International (Louisville, KY).

EGTA, HEPES and BSA were purchased from Sigma Chemical Co, (St. Louis, MO). [¹²⁵I] - Bolton-Hunter (BH-CCK₈)

(specific activity, 2200 Ci/mmol) was obtained from New England Nuclear (Boston, MA). Male guinea pigs, 250 to 325 g, were obtained from Scientific Small Animal Laboratory and Farm (Arlington Heights, IL). Collagenase, code CLSPA was purchased from Worthington (Frehold, New Jersey)

Protocol For Radioligand Binding Experiments in Guinea Pig Cerebral Cortical and

Pancreatic Membrane Preparations

Cortical and pancreatic membranes were prepared as described (Lin and Miller; J. Pharmacol. Exp. Ther. 232, 775-780, 1985). In brief, cortex and pancreas were removed and rinsed with ice-cold saline. Visible fat and

connective tissues were removed from the pancreas. Tissues were weighed and homogenized separately in approximately 25 mL of ice-cold 50 mM Tris-HCl buffer, pH 7.4 at 4°C. with a Brinkman Polytron for 30 sec, setting 7. The homogenates were centrifuged for 10 min at 1075 x g and pellets were discarded. The supernatants were saved and centrifuged at 38,730 × g for 20 min. The resultant pellets were

rehomogenized in 25 mL of 50 mM Tris-HCl buffer with a Teflon-glass homogenizer, 5 up and down strokes. The homogenates were centrifuged again at 38,730 × g for 20 min. Pellets were then resuspended in 20 mM HEPES,

containing 1 mM EGTA, 118 mM NaCl, 4.7 mM KCl, 5 mM MgCl₂, 100 uM bestatin, 3 uM phosphoramidon, pH 7.4 at 22°C. with a Teflon-glass homogenizer, 15 up and down strokes.

Resuspension volume for the cortex was 15-18 mL per gm of original wet weight and 60 mL per gm for the pancreas.

Incubation Conditions [ ¹²⁵I] Bolton-Hunter CCK₈ and test compounds were diluted with HEPES-EGTA-salt buffer (see above) containing

0.5% bovine serum albumin (BSA). To 1 mL Skatron

polystyrene tubes were added 25 uL of test compounds, 25 uL of [ ¹²⁵I]BH-CCK₈ and 200 uL of membrane suspension. The final BSA concentration was 0.1%. The cortical tissues were incubated at 30°C for 150 min. and pancreatic tissues were incubated at 37°C for 150 min. Incubations were terminated by filtration using Skatron Cell Harvester and

SS32 microfiber filter mats. The specific binding of

[ ¹²⁵I]BH-CCK₈, defined as the difference between binding in the absence and presence of 1 uM CCK₈, was 85-90% of total binding in cortex and 90-95% in pancreas. IC₅₀s were determined from the Hill analysis. The results of these binding assays are shown in Table 1.

Table 1

Compound of ¹²⁵ I-BH- CCK₈ ¹²⁵I-BH-CCK₈

Example Pancreas Cortex

12 30 270

16 12 680

17 10 732

31 26 238

33 71 1480

35 26 1800

39 32 114 45 35 4700

47 50 4000

49 41 815

53 22 3400

57 6 - - - -

73 5.2 970

157 3.0 1450

180 2.5 3470

193 7 - - - -

212 79 10000

214 14 5000

215 48 2000

226 13 4000

229 6 3000

232 4 10000

233 4 10000

236 30 - - -

The results indicate that compounds of the invention possess selective affinity for the pancreatic CCK

receptors.

2. Protocol for Amylase Release Assay

This assay was performed using the modified protocol of Lin et al., J. Pharmacol. Exp. Ther. 236, 729-734, 1986.

Guinea Pig Acini Preparation

Guinea pig acini were prepared by the method of

Bruzzone et al. (Biochem. J. 226, 621-624, 1985) as

follows. Pancreas was dissected out and connective tissues and blood vessels were removed. The pancreas was cut into small pieces (2mm) by a scissor and placed in a 15 mL conical plastic tube containing 2.5 mL of Krebs-Ringer HEPES (KRH) buffer plus 400 units per mL of collagenase. The composition of the KRH buffer was: HEPEs, 12.5 mM: NaCl, 118 mM; KCl, 4.8 mM; CaCl₂, 1 mM; KH₂PO₄, 1.2 mM; MgSO₄, 1.2 mM; NaHCO₃, 5 mM; glucose, 10 mM, pH 7.4. The buffer was supplemented with 1% MEM vitamins, 1% MEM amino acids and 0.001% aprotinin. The tube was shaken by hand until the suspension appeared homogeneous, usually 5 to 6 min. 5 mL of the KRH, without collagenase and with 0.1% BSA, were added and the tube was centrifuged at 50 × g for 35 sec. The supernatant was discarded and 6 mL of the KRH were added to the cell pellet. Cells were triturated by a glass pipet and centrifuged at 50 × g for 35 sec. This wash procedure was repeated once. The cell pellet from the last centrifugation step was then resuspended in 15 mL of KRH containing 0.1% BSA. The contents were filtered through a dual nylon mesh, size 275 and 75 um. The filtrate, containing the acini, was centrifuged at 50 × g for 3 min. The acini were then resuspended in 5 mL of KRH-BSA buffer for 30 min at 37°C. under 100% O₂, with a change of fresh buffer at 15 min.

Amylase Assay

After the 30 min incubation time, the acini was resuspended in 100 volumes of KRH-BSA buffer, containing 3 uM phosphoramidon and 100 uM bestatin. While stirring, 400 uL of acini were added to 1.5 mL microcentrifuge tubes containing 50 uL of CCK₈, buffer, or test compounds. The final assay volume was 500 uL. Tubes were vortexed and placed in a 37°C waterbath, under 100% O₂, for 30 min.

Afterward, tubes were centrifuged at 10,000 g for 1 min.

Amylase activity in the supernatant and the cell pellet were separately determined after appropriate dilutions in

0.1% Triton X-100, 20 mM NaH₂PO₄, pH 7.4 by Abbott Amylase

A-gent test using the Abbott Bichromatic Analyzer 200. The reference concentration for CCK₈ in determining the EC₅₀s of the compounds of Formula I was 3 × 10^-10M. The results of this assay are shown in Table 2.

TABLE 2

Compound of Example Amylase release, EC₅₀(nM)

16 5

17 3

31 40

33 80

39 24

157 1.1

180 0.74

The results indicate that compounds of the invention are CCK agonists.

The ability of the compounds of the invention to increase the release of insulin in vivo can be demonstrated using the following protocol.

Measurement of Plasma Insulin in Mice Following

Treatment With CCK₈ or a CCK Aσonist

Male mice, 20-30 g, were used in all experiments. The animals were fed with laboratory lab chow and water ad libitum. CCK₈ or the CCK agonist compound of this

invention was injected into the tail vein. Two minutes later, the animals were sacrificed and the blood was collected in 1.5 mL heparinized polypropylene tubes. The tubes were centrifuged at 10,000 × g for 2 minutes. The insulin levels were determined in the supernatant, i.e., plasma, by RIA using kits obtained from Radioassay Systems Laboratory (Carson, CA.) or Novo Biolabs (Danbury, CT.). The results of this assay are shown in Table 3 and are expressed as the percent increase in insulin secretion over insulin secretion observed in mice injected with a saline solution. Each dose was tested in at least six mice and the values presented are averages for the group of mice tested at each dose.

Table 3

Effect of CCK Aσonists On Insulin Secretion in Mice

% Increase In Insulin

Dose Secretion versus

Compound of Example (nmole/kg) Saline Control

157 10 41

100 112

180 100 238

CCK₈ 3 65

10 85

30 90

100 70

The results indicate that compounds of the invention stimulate insulin secretion in vivo.

The ability of the compounds of the invention to modulate central nervous system function in vivo can be demonstrated using the following protocol.

Behayioral Effect of CCK Agonists in Mice

Male Swiss CD-1 mice (Charles River) (22-27 g) are provided ample food (Purina Lab Chow) and water until the time of their injection with the test compounds.

I.p. injections are given as a volume of 10.0 mL/kg using a 26 gauge, 3/8 inch needle. ICV injections were given by a free-hand method similar to that previously described (Haley and McCormick, Br. J. Pharmacol.

Chemother., 12 12-15 (1957)). The animals were placed on a slightly elevated metal grid and restrained by the thumb and forefinger at the level of the shoulders, thus

immobilizing their heads. Injections were made with a 30 gauge needle with a "stop" consisting of a piece of tygon tubing to limit penetration of the needle to about 4.5 mm below the surface of the skin. The needle was inserted perpendicular to the skull at a midline point equidistant from each eye and an equal distance posterior from the level of the eyes such that the injection site and the two eyes form an equilateral triangle. The injection volume (5 ul) was expelled smoothly over a period of approximately 1 second.

Immediately after the injections the mice were placed in their cages and allowed a 15 minute recovery period prior to the beginning of behavioral observations.

For the behavioral observations, the mice were placed in clear plastic cages. Each cage measured 19×26×15 cm and contained a 60-tube polypropylene test tube rack place on end in the center of the cage to enhance exploratory activity. Observations were made every 30 seconds for a period of 30 minutes. Behavior was compared between drug treated mice and mice treated with an equal volume of carrier (usually 0.9% saline or 5% dimethylsulfoxide in water). Locomotion as reported here consisted of either floor locomotion or active climbing on the rack.

Differences among groups were analyzed by Newman-Kewels analysis and a probability level of p<.05 was accepted as significant. Each group tested consisted of 10 animals . The results of this test are shown in Table 4 and Table 5.

Table 4

Suppression of Locmotor Activity in Mice Following

IP Administration of CCK Agonists compound of Example Minimal Effective Dose

CCK₈ 0.001 micromol/kg

106 1.0 micromol/kg

157 0.03 micromol/kg

180 0.01 micromol/kg

Table 5

Suppression of Locomotor Activity in Mice Following

ICV Administration of CCK Agonists

Compound of Example Minimal Effective Dose

CCK₈ 3.0 nmol/mouse

106 10.0 nmol/mouse

157 30.0 nmol/mouse

180 1.0 nmol/mouse

The results of these tests indicate that compounds of the invention suppress locomotor activity and thus

demonstrate pyschoactive properties.

The ability of the compounds of the invention to suppress feeding can be demonstrated using the following protocols. Feeding Effect of CCK Agonists in Rats

Forty male, Sprague-Dawley rats were subjected to a 23 hour food deprivation schedule for four days. On the fifth day, the animals were divided into five equal groups based on their previous (4th day) food intake. Five minutes prior to their one hour free feeding (Purina Rat Chow), the animals were injected (i.p.) with either vehicle, CCK₈ or the compound of Example 106. The amount of food consumed was measured after subtraction of spillage. The results of this test are shown in Table 6.

Table 6

Suppression of Feeding in Rats Following I.P.

Administration of CCK Agonists

Compound Dose Mean Food Intake

vehicle --- 9.40 grams

CCKg 20 ug/kg 6.56 grams

Example 106 1.0 mg/kg 3.49 grams

Example 106 3.0 mg/kg 1.80 grams

Example 106 9.0 mg/kg 1.90 grams

The results of this test indicate that compounds of the invention reduce food intake.

Feeding Effects of Chronic Administration of CCK-8 and

CCK Agonists in Rat.

Adult male Sprague-Dawley rats, weighing approximately 250 g, were placed on a restricted diet. Rats were weighed every morning and were allowed access to a liquid diet (Ensure) for 60 minutes in the morning and 45 minutes in the late afrernoon; intakes were recorded every 15 minutes. Following an 8-day diet acclimation period, the rats were weighed and injected with vehicle, CCK-8 (10 nmol/kg), or compound of Example 180 (1 or 10 nmol/kg) (ip) . Six to 10 minutes later, the rats were presented with the diet, and intakes were recorded as usual. No injections were administered prior to the PM feeding.

After 11 days of testing, half of the animals were

sacrificed while the remaining rats were withdrawn from treatment and injected with vehicle for 4 days.

Graphs of AM food intakes and body weight changes for the various treatment groups are shown in Figs. 1 and 2, respectively (there was no difference between groups in PM intakes so these data are not shown). Statistical analysis indicated that CCK-8 significantly reduced intakes on just the first and last treatment days and produced no

significant effects on body weight gain. In contrast, the compound of Example 180 (1 nmol/kg) significantly reduced intakes on days 1,2,3,5,6,7 and 11 and animals in this groups showed significantly less weight gain than controls on days 7 and 8 of testing. Rats given the higher dose of the compound of Example 180 (10 nmol/kg) showed

significantly reduced intakes on every drug injection day, and their rate of body weight gain also differed significantly from that of controls across the entire testing period.

The results of this test indicate that compounds of the invention reduce food intake when administered

chronically.

The compounds of the present invention can be used in the form of salts derived from inorganic or organic acids. These salts include but are not limited to the following: acetate, adipate, alginate, citrate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, camphorate,

camphorsuIfonate, digluconate, cyclopentanepropionate, dodecylsulfate, ethanesulfonate, glucoheptonate,

glycerophosphate, hemisulfate, heptonate, hexanoate, fumarate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxy-ethanesulfonate, lactate, maleate,

methanesulfonate, nicotinate, 2-naphthalenesulfonate, oxalate, pamoate, pectinate, persulfate, 3-phenylpropionate, picrate, pivalate, propionate, succmate, tartrate, thiocyanate, tosylate, and undecanoate. Also, the basic nitrogen-containing groups can be quaternized with such agents as loweralkyl halides, such as methyl, ethyl, propyl, and butyl chloride, bromides, and iodides; dialkyl sulfates like dimethyl, diethyl, dibutyl, and diamyl sulfates, long chain halides such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides, aralkyl halides like benzyl and phenethyl bromides, and others. Water or oil-soluble or dispersible products are thereby obtained.

The pharmaceutically acceptable salts of the present invention can be synthesized from the compounds of formula I which contain a basic or acidic moiety by conventional chemical methods. Generally, the salts are prepared by reacting the free base or acid with stoichiometric amounts or with an excess of the desired salt forming inorganic or organic acid or base in a suitable solvent or various combinations of solvents.

Examples of acids which may be employed to form pharmaceutically acceptable acid addition salts include such inorganic acids as hydrochloric acid, sulphuric acid and phosphoric acid and such organic acids as oxalic acid, maleic acid, succinic acid and citric acid. Other salts include salts with alkali metals or alkaline earth metals, such as sodium, potassium, calcium or magnesium or with organic bases.

The pharmaceutically acceptable salts of the acids of formula I are also readily prepared by conventional procedures such as treating an acid of formula I with an appropriate amount of a base, such as an alkali or alkaline earth metal hydroxide e.g. sodium, potassium, lithium, calcium, or magnesium, or an organic base such as an amine, e.g., dibenzylethylenediamine, cyclohexylamine,

dicyclohexylamine, trimethylamine, piperidine, pyrrolidine, benzylamine and the like, or a quaternary ammonium

hydroxide such as tetramethylammonium hydroxide and the like.

When a compound of formula I is used as an agonist of CCK or gastrin in a human subject, the total daily dose administered in single or divided doses may be in amounts, for example, from 0.001 to 1000 mg a day and more usually 1 to 1000 mg. Dosage unit compositions may contain such amounts of submultiples thereof to make up the daily dose. The amount of active ingredient that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated, the particular treatment and the particular mode of administration.

It will be understood, however, that the specific dose level for any particular patient will depend upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, rate of excretion, drug

combination, and the severity of the particular disease undergoing therapy.

The compounds of the present invention may be

administered orally, parenterally, by inhalation spray, rectally, or topically in dosage unit formulations

containing conventional nontoxic pharmaceutically

acceptable carriers, adjuvants, and vehicles as desired. Topical adminsitration may also involve the use of

transdermal administration such as transdermal patches or iontophoresis devices. The term parenteral as used herein includes subcutaneous injections, intravenous,

intramuscular, intrasternal injection, or infusion

techniques.

Injectable preparations, for example, sterile

injectable aqueous or oleagenous suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable prepartion may also be a sterile injectable solution or suspension in a nontoxic

parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water. Ringer's solution, and isotonic sodium chloride solution. In additon, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.

Suppositories for rectal administration of the drug can be prepared by mixing the drug with a suitable

nonirritating excipient such as cocoa butter and

polyethylene glycols which are solid at ordinary

temperatures but liquid at the rectal temperature and will therefore melt in the rectum and release the drug.

Solid dosage forms for oral administration may include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound may be admixed with at least one inert diluent such as sucrose lactose or starch. Such dosage forms may also comprise, as is normal practice, additional substances other than inert diluents, e.g., lubricating agents such as magnesium stearate. In the case of capsules, tablets, and pills, the dosage forms may also comprise buffering agents. Tablets and pills can additionally be prepared with enteric coatings.

Liquid dosage forms for oral administration may include pharmaceutically acceptable emulsion, solutions, suspensions, syrups, and elixirs containing inert diluents commonly used in the art, such as water. Such compositions may also comprise adjuvants, such as wetting agents, emulsifying and suspending agents, and sweetening,

flavoring, and perfuming agents.

The present agents can also be administered in the form of liposomes. As is known in the art, liposomes are generally derived from phospholipids or other lipid

substances. Liposomes are formed by mono- or multi-lamellar hydrated liquid crystals that are dispersed in an aqueous medium. Any non-toxic, physiologically acceptable and metabolizable lipid capable of forming liposomes can be used. The present compositions in liposome form can contain, in addition to the tetrapeptide of the present invention, stabilizers, preservatives, excipients, and the like. The preferred lipids are the phospholipids and the phosphatidyl cholines (lecithins), both natural and synthetic.

Methods to form liposomes are known in the art. See, for example, Prescott, Ed., Methods in Cell Biology. Volume XIV, Academic Press, New York, N.Y. (1976), p. 33 et seq.

The foregoing is merely illustrative of the invention and is not intended to limit the invention to the disclosed compounds. Variations and changes which are obvious to one skilled in the art are intended to be within the scope and nature of the invention which are defined in the appended claims.

Claims

CLAIMS What is claimed is :

1. A compound of the formula:

Y - X - Z - Q

wherein Y is

1)

or

2)

wherein

(a) A is an N-protecting group and

B is -N(R₂₁)- wherein R₂₁ is hydrogen or loweralkyl; or wherein A is absent and B is halogen, cyano, or -OR₂₂ wherein R₂₂ is hydrogen, alkanoyl,

alkoxycarbonyl or loweralkyl;

U is -CH-;

L is -CH₂-; R₁ is bicyclic carbocyclic or bicyclic heterocyclic; and

R₂₀ is -C(O)- or -CH₂-;

or

(b) A and B are absent;

U and L taken together are an alkylene or alkenylene group;

R₁ is bicyclic carbocyclic or bicyclic heterocyclic; and

R₂₀ is -C(O)- or -CH₂-;

or

(c) A is absent;

B is absent or when U is N then B is hydrogen,

loweralkyl or -(CH₂)_xOR₂₃ wherein x is 2-6 and

R₂₃ is hydrogen, loweralkyl, aryl or heterocyclic;

U is N, O or S;

L is -CH₂;

R₁ is bicyclic carbocyclic or bicyclic heterocyclic; and

or

(d) A is alkoxy, acyloxy, bicyclic carbocyclic or

bicyclic heterocyclic;

B is an alkylene group;

U is CH;

L is an alkylene group;

R₁ is alkoxy, acyloxy, bicyclic carbocyclic or bicyclic heterocyclic;

and R₂₀ is -C(O)- or -CH₂-; or

(e) A and B are absent;

U is -CH₂-;

L is O, S or -N(R₂₆)- wherein R₂₆ is hydrogen or

loweralkyl;

R is bicyclic carbocyclic or bicyclic heterocyclic, and

R₂₀ is -C(O)- or -CH₂-;

X is

wherein V is C₁ to C₆ alkylene or C₂ to C₆

alkenylene;

R₂₇ is hydrogen or loweralkyl;

T is -C(O)- or -CH₂-; is O, S, -N(R₃₀)- wherein R₃₀ is hydrogen or

loweralkyl, or E is -CH₂C (O) -, -C(O)CH₂-,

-CH₂CH₂-, -CH=CH-, -C (=R₂₁ ) R₂₃-, -R₂₂C (=R₂₁ ) or

-R₂₃C(=R₂₄)R₂₃- wherein R₂₁ is O or S, R₂₂ is

N(R₂₈), O or s, R₂₃ is independently selected at each occurrence from N(R₂₉), O and S, and R₂₄ is O, S or NH, wherein R₂₈ and R₂₉ are

independently selected from hydrogen and loweralkyl; with the proviso that when R₂₂ is NH and V is alkenylene then the NH is not directly bonded to the double bond of the alkenylene group; and

R₂ is -GR₇ wherein G is absent, C₁ to C₄ alkylene, C₂ to C₄ alkenylene or a C₁ to C₄ alkylene or C₂ to C₄ alkenylene group which is substituted by a cyano group or an N-protected amino group and R₇ is loweralkyl, C₁ to C₁₂ alkenyl, adamantyl, aryl, arylalkyl, heterocyclic, cycloalkyl, substituted loweralkyl wherein the loweralkyl group is substituted with alkoxy, thioalkoxy, halo or N-protected amino or R₇ is substituted cycloalkyl wherein the cycloalkyl ring is substituted with one to four substituents independently selected from loweralkyl, halo and alkoxy; with the proviso that when E is

-NHC(O)NH- then G is absent and with the proviso that when R₂₃ is NH and G is C₂ to C₄ alkenylene then the NH is not directly bonded to the double bond of the alkenylene group;

Z is

or

R₃₆ is hydrogen or loweralkyl; R₂₅ is -C(O)- or -CH₂-; and R₈₁ is -OR₈₂ wherein R₈₂ is hydrogen or loweralkyl or R₈₁ is

Q is

wherein R₄ is cyclohexyl, loweralkyl, aryl,

arylalkyl, heterocyclic or (heterocyclic) alkyl;

Rr is hydrogen or loweralkyl;

R₃₃ is -C(O)-, -C(S)- or -CH₂-; and

D is -NR₃₄R₈₀ wherein R₃₄ is hydrogen, hydroxy or

loweralkyl and Rso is hydrogen or loweralkyl, or D is -SR₃₅ wherein R₃₅ is hydrogen or loweralkyl, or D is -OR₃₆ wherein R₃₆ is

hydrogen, loweralkyl or alkanoyl;

or a pharmaceutically acceptable salt thereof.

2. The compound of Claim 1 wherein Y is t-butoxycarbonyl-Trp, t-butoxycarbonyl-D-Trp,

t-butoxycarbonyl-beta-Nal, acetyl-Trp or indolelactoyl;

Z is Asp or N-Me-Asp;

X is

wherein E is -NHC (O)- or -NHC(O)NH- and R₂ is aryl or

X is

wherein R₂ is aryl; and

Q is Phe-NH₂, N-Me-Phe-NH₂, alpha-Nal-NH₂ or phenlyalaninol.

3. A compound selected from the group consisting of: t-butoxycarbonyl-beta-Nal-Lys (epsilon-N-(4-hydroxycinnamoyl))-Asp-Phe-NH₂; t-butoxycarbonyl-Trp-Lys(epsilon-N-(4- hydroxycinnamoyl))-Asp-(NMe)Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(6-hydroxy-2- naphthoyl))-Asp-(NMe)Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(6-acetoxy-2- naphthoyl))-Asp-(NMe)Phe-NH₂;

t-butoxycarbonyl-D-Trp-Lys(epsilon-N-(4-hydroxycinnamoyl))-Asp-(NMe)Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-((6-OSO₃H-2-naρhthoyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(5-hydroxyindole-2-carbonyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-(1-naphthylaminocarbonyl))-Asp-Phe-NH₂;

t-butoxycarbonyl-Trp-Lys(epsilon-N-4-hydroxycinnamoyl)ψ(CH₂NH)Asp-Phe-NH₂;

BOC-Trp-Lys(ε-N-[2-bromophenylaminocarbonyl])-Asp-Phe-NH2; BOC-Trp-Lys(ε-N-[2-methylphenylaminocarbonyl])-Asp- Phenylalaninol;

Indolelactoyl-Lys(ε-N-[2-methylphenylaminocarbonyl])-Asp-Phe-NH₂;

Trp-Lys(ε-N-[2-methylphenylaminocarbonyl])-Asp-(NMe)Phe-NH₂

Hydrochloride; Ac-Trp-Lys (ε-N- [2-methylphenylaminocarbonyl] ) -Asp-Phe-NH_2; BOC-Trp-Lys (ε-N- [2-methylphenylaminocarbonyl] ) - (NMe) Asp-Phe -NH_{2 ;}

BOC-Trp-Lys (ε-N- [2-methylphenylaminocarbonyl] ) - (NMe) Asp- (NMe) Phe-NH2 ; and

BOC-Trp-Lys(ε-N-[2-methylphenylaminocarbonyl])[ψCH₂NH]-Asp-Phe-NH₂;

or a pharmaceutically acceptable salt thereof.

4. BOC-Trp-Lys(epsilon-N-(2-methylphenylaminocarbonyl))-Asp-(NMe) Phe-NH₂, or a pharmaceutically

acceptable salt thereof.

5. A method for mimicking the effects of CCK on CCK receptors comprising administering to a host in need of such treatment a therapeutically effective amount of a compound of Claim 1.

6. A method for (a) stimulating insulin secretion, (b) inducing central nervous system suppression, (c) treating psychosis, or (d) treating appetite disorders comprising administering to host in need of such treatment a therapeutically effective amount of a compound of Claim 1.

7. A CCK agonist composition comprising a

pharmaceutical carrier and a therapeutically effective amount of a compound of Claim 1.

8. A composition for (a) stimulating insulin

secretion, (b) inducing central nervous system suppression, (c) treating psychosis, or (d) treating appetite disorders comprising a pharmaceutical carrier and a therapeutically effective amount of a compound of Claim 1.

9. A process for the preparation of a compound of Claim 1 wherein R₂₀, T and R₂₅ are -C(O)- comprising:

(a) stepwise coupling of the appropriately protected amino acids or amino acid analogs represented by Y, X, Z and Q as defined herein, followed by removal of the protecting groups; or

(b) coupling of the appropriately protected fragments of dipeptide length or greater, said fragments comprising the amino acids or amino acid analogs represented by Y, X, Z and Q as defined herein, followed by removal of the

protecting groups.

10. A process for the preparation of a compound of Claim 1 wherein R₂₀, T and R₂₅ are -C(O)- and wherein E is O, S, -N(R₃₀)- wherein R30 is hydrogen or loweralkyl, or E is -R₂₂C(=R₂₁)- or -R₂₃C (=R₂₄) R₂₃- wherein R₂₁ is O or S, R₂₂ is N(R₃₀), O or S, R₂₃ is independently selected at each occurrence from N(R₃₀), O and S; and R₃₀ is independently selected at each occurrence from hydrogen and loweralkyl comprising:

(a) stepwise coupling of the appropriately protected amino acids or amino acid analogs represented by Y,

Z and Q as defined herein, and wherein P' is hydrogen or a protecting group, followed by functionalization of the side chain of the amino acid analog represented by X and removal of the protecting groups; or

(b) coupling of the appropriately protected fragments of dipeptide length or greater, said fragments comprising the amino acids or amino acid analogs represented by Y,

Z and Q as defined herein, and wherein P' is hydrogen or a protecting group, followed by functionalization of the side chain of the amino acid analog represented by X and removal of the protecting groups.

11. A process for the preparation of a compound of Claim 1 wherein one or more of the amide bonds between the amino acids or amino acid analogs represented by Y, X, Z and Q as defined herein is in the form of a reduced carbonyl amide bond surrogate such that one or more of R₂₀, T and R₂₅ is -CH₂-, comprising for each reduced carbonyl amide bond surrogate:

(a) condensing the appropriately protected amino aldehyde derived from the corresponding amino acid or amino acid analog represented by Y, X, Z and Q with the free terminal amino group of the adjacent appropriately protected amino acid, amino acid analog, dipeptide, dipeptide analog, tripeptide or tripeptide analog, followed by;

(b) reduction of the resulting Schiff base and removal of the protecting groups.

12. A process for the preparation of a compound of Claim 1 wherein one or more of the amide bonds between the amino acids or amino acid analogs represented by Y, X, Z and Q as defined herein is in the form of a reduced

carbonyl amide bond surrogate such that one or more of R₂₀, T and R₂₅ is -CH₂-, and wherein E is O, s, -N(R₃₀)- wherein R₃₀ is hydrogen or loweralkyl, or E is -R₂₂C(=R₂₁) - or

-R₂₃C (=R₂₄)R₂₃- wherein R₂₁ is 0 or S, R₂₂ is N(R₃₀), 0 or S, R₂₃ is independently selected at each occurrence from

N(R₃₀), 0 and S; and R₃₀ is independently selected at each occurrence from hydrogen and loweralkyl comprising for each reduced carbonyl amide bond surrogate:

(a) condensing the appropriately protected amino aldehyde derived from the corresponding amino acid or amino acid analog represented by Y,

Z and Q as defined herein, and wherein P' is hydrogen or a protecting group, with the free terminal amino group of the adjacent appropriately protected amino acid, amino acid analog, dipeptide, dipeptide analog, tripeptide or

tripeptide analog, followed by;

(b) reduction of the resulting Schiff base,

functionalization of the side chain of the amino acid analog represented by X and removal of the protecting groups.