AU2001296313B2 - Melanocortin receptor ligands - Google Patents

Description

WO 02/26774 PCT/US01/30051 MELANOCORTIN RECEPTOR LIGANDS FIELD OF THE INVENTION The present invention relates to new melanocortin (MC) receptor ligands. These ligands preferably exhibit selectivity for the MC-4 and/or the MC-3 receptors relative to the other melanocortin receptors (in particular the MC-1 receptor).

BACKGROUND OF THE INVENTION Melanocortin peptides (melanocortins) are natural peptide horihones in animals and man that bind to and stimulate MC receptors. Examples of melanocortins are a-MSH (melanocyte stimulating hormone), P-MSH, y-MSH, ACTH (adrenocorticotropic hormone) and their peptide fragments. MSH is mainly known for its ability to regulate peripheral pigmentation (Eberle 1988), whereas ACTH is known to induce steroidoneogenesis (Simpson and Waterman, 1988).

The melanocortin peptides also mediate a number of other physiological effects. They are reported to affect motivation, learning, memory, behavior, inflammation, body temperature, pain perception, blood pressure, heart rate, vascular tone, natriuresis, brain blood flow, nerve growth and repair, placental development, aldosterone synthesis and release, thyroxin release, spermatogenesis, ovarian weight, prolactin and FSH secretion, uterine bleeding in women, sebum and pheromone secretion, sexual activity, penile erection, blood glucose levels, intrauterine fetal growth, food motivated behavior, as well as other events related to parturition.

ACTH and the various MSH peptides share the tetrapeptide core His-Phe-Arg-Trp. All of the peptides are derived from the proteolytic processing of the pro-peptide preopiomelanocortin (POMC). In the past several years, five distinct melanocortin receptor subtypes have been identified. These MC receptors belong to the class of 7 transmembrane domain G-protein coupled receptors. The five MC receptors, termed MC-1, MC-2, MC-3, MC-4 and MC-5, all couple in a stimulatory fashion to cAMP. Of these, the MC-2 receptor is the ACTH receptor, whereas the others constitute subtypes of MSH receptors. The MC-1 receptor is present on melanocytes and melanoma. The MC-2 receptor is present predominantly in the adrenal gland. The mRNA for the MC-3 receptor has been found in the brain, as well as in placental and gut tissues (Gantz et al. 1993a, Desaraud et al. 1994, Roselli Rehfuss et al. 1993).

The MC-4 receptor has been found primarily in the brain (Gantz et al. 1993b; Mountjoy et al 1994). The MC-5 receptor is expressed in the brain, as well as in several peripheral tissues WO 02/26774 PCT/US01/30051 (Chhajlani et al 1993; Gantz et al 1994; Griffon et al 1994; Labbu et al. 1994; Barrett et al. 1994; Fathi et al. 1995). More recent data from humans indicate that all of the cloned MC-receptors have a wider tissue distribution (Chhajlani, 1996) than originally thought.

As discussed above, the members of the melanocortin receptor family can be a differentiated on the basis of their tissue distribution. Both the MC-4 and MC-3 receptors have been localized to the hypothalamus, a region of the brain believed to be involved in the modulation of feeding behavior. Compounds showing selectivity for the MC-4/MC-3 receptors have been shown to alter food intake following intracerebroventricular and peripheral injection in rodents. Specifically, agonists have been shown to reduce feeding, while antagonists have been shown to increase feeding. See, Fan, W. et al., "Role of Melanocortinergic Neurons in Feeding and the Agouti Obesity Syndrome", Nature, 385(6612), pp. 165-8 (Jan. 9, 1997).

The role of the MC-4 receptor subtype has been more clearly defined in the control of eating and body weight regulation in mammals. See, Huszer, D. et al., "Targeted Disruption of the Melanocortin-4 Receptor Results in Obesity in Mice", Cell, Vol. 88, pp. 131-141 (1997); Klebig, M.L. et al., "Ectopic Expression of the Agouti Gene in Transgenic Mice Causes Obesity, Features of Type II Diabetes, and Yellow Fur", Proc. Natl Acad Sci., Vol. 92, pp. 4728-32 (1995); Karbon, W. et al., "Expression and Function of Argt, a Novel Gene Related to Agouti", Abstract from the Nineteenth Annual Winter Neuropeptide Conference (1998); Fan, W. et al., "Role of Melanocortinergic Neurons in Feeding and the Agouti Obesity Syndrome", Nature, Vol.

385, pp. 165-168 (1997); Seely, "Melanocortin Receptors in Leptin Effects", Nature, Vol.

390, p. 349 (1997); Comuzzie, "A Major Quantitative Trait Locus Determining Serum Leptin Levels and Fat Mass is Located on Human Chromosome Nat. Gen., Vol. 15, pp. 273- 276 (1997); Chagnon, Y.C. et al., "Linkage and Association Studies Between the Melanocortin Receptors 4 and 5 Genes and Obesity-Related Phenotypes in the Quebec Family Study", Mol.

Med., Vol 3(10), pp. 663-673 (1997); Lee, F. and Huszar, D, "Screening Methods for Compounds Useful in the Regulation of Body Weight", World Patent Publication WO 97/47316 (1997); and Shutter, J.R. et al., "Hypothalamic Expression of ART, a Novel Gene Related to Agouti, is Up- Regulated in Obese and Diabetic Mutant Mice", Gen. Dev. Vol. 11, pp. 593-602 (1997).

Stimulation of the MC-4 receptor by its endogenous ligand, aMSH, produces a satiety signal and may be the downstream mediator of the leptin satiety signal. It is believed that by providing potent MC-4 receptor agonists, appetite may be suppressed and weight loss benefits may be achieved.

0 The role of the MC3 receptor subtype has recently been defined in the control of body r, weight regulation and energy partitioning. See, Chen, A.S. et al., Inactivation of the mouse S melanocortin-3 receptor results in increased fat mass and reduced lean body mass", Nature Genetics, Vol 26, pp 97-102 (2000); Butler, A.A. et al., "A Unique Metabolic Syndrome Causes Obesity in the Melanocortin-3 Receptor -Deficient Mouse", Endocrinology, Vol 141, pp 3518- 3521 (2000). is believed that MC-3 receptor agonists may modulate energy partitioning and c weight loss benefits may be achieved.

These studies imply a non-redundant role for the MC-3 receptor, compared to MC-4 OC receptor, in energy homeostasis. Therefore, compounds that stimulate both MC-3 and MC-4 receptors may enhance the weight loss benefits compared to compounds that are selective for either the MC-3 or MC-4 receptor subtypes.

The Applicants have discovered a class of compounds that surprisingly have high affinity for the MC-4 and/or the MC-3 receptor subtypes, and that are typically selective for these MC receptors relative to the other melanocortin receptor subtypes, particularly the MC-1 subtype. It is therefore an aspect of this invention to provide compounds that have affinity for the MC-4 and/or MC-3 receptor subtypes. It is a further aspect of the invention to provide means for administration of said compounds to animals or man. Still other aspects of the invention will be evident from the following disclosure of the invention.

The discussion of documents, acts, materials, devices, articles and the like is included in this specification solely for the purpose of providing a context for the present invention. It is not suggested or represented that any or all of these matters formed part of the prior art base or were common general knowledge in the field relevant to the present invention as it existed before the priority date of each claim of this application.

Throughout the description and the claims of this specification the word "comprise" and variations of the! word, such as "comprising" and "comprises" is not intended to exclude other additives, components, integers or steps.

DISCLOSURE OF THE INVENTION The invention relates to a class of compounds that are ligands for receptors of the MC-4 and/or the MC-3 subtype. In particular, the invention relates to a compound having a structure according to Formula (dR 4

('R

6

R

6 R(dR9 1 j 3 42 wherein R I WO 02/26774 PCT/US01/30051 X is selected from hydrogen, fluoro, aryloxy, acyloxy, OW', SRI, -NR'R' and -CHR'R', where R 1 and R are independently selected from the group consisting of hydrogen, alkyl and acyl; each R 2 is independently selected from the group consisting of hydrogen, alkyl halo and heteroalkyl; or two consecutive R 2 moieties, or consecutive R 2 and R 3 moieties, may join to form a 3 to 8 membered carbocyclic or heterocyclic ring; or the R 2 bonded to the carbon atom that is bonded to X and Z 1 and an R 5 moiety can optionally join to form a carbocyclic or heterocyclic ring that is fused to phenyl ring J; or the R 2 bonded to the carbon atom that is bonded to ring Ar can join with an R 7 to form a ring fused to ring Ar; or the R 2 bonded to the carbon atom that is bonded to Z 2 and Z 3 can optionally join with R 8 to form a carbocyclic or heterocyclic ring; or the R 2 bonded to the carbon atom that is bonded to Z 3 and D can optionally join with R'O to form a carbocyclic or heterocyclic ring; each of Z 1

Z

2 and Z 3 is independently selected from -OC(R 3

)(R

3 -C(R3)(R 3 S(O)aC(R)(R 3 where a is 0, 1 or 2; -C(R 3

)(R

3 where b is 0, 1 or 2; N(R )C(R3)(R -C(R -N(R 3

C(R

3 -C(R3)=C(R 3

-SO

2 N(R3d)-; N(R3d)SO 2

-C(R

3 )(R3)P(=O)(OR 3

-P(=O)(OR)C(R

3

)(R

3

-N(R

3 3 f)N(R3d)-; a cycloalkyl having from 3 to 8 ring atoms and a heterocycloalkyl having from 4 to 8 ring atoms; wherein each of R 3

R

3a

R

3 b and R 3 c, when present, is independently selected from hydrogen, hydroxy, alkoxy, aryloxy, acyloxy, thiol, alkylthio, acylthio, arylthio, amino, alkylamino, acylamino, and alkyl;

R

3 d, when present, is selected from hydrogen, alkyl and aryl;

R

3 when present, is selected from hydrogen, alkyl, aryl and acyl; and

R

f when present, is selected from hydrogen and alkyl; p is 0, 1, 2, 3, 4 or 5; wherein when p is greater than 0, each R 4 and R 4 is independently selected from hydrogen, alkyl, aryl, halo, hydroxy, alkoxy, amino and acylamino; WO 02/26774 PCT/US01/30051 when p is greater than 1, two R 4 moieties, together with the carbon atoms to which they are bonded, can join to form a heterocycloalkyl, cycloalkyl or aryl ring; and when p is greater than 1, the R 4 moieties on two adjacent carbon atoms can both be nil such that a double bond is formed between the two adjacent carbon atoms, or both the R 4 and R 4 moieties on two adjacent carbon atoms can all be nil such that a triple bond is formed between the two adjacent carbon atoms;

R

5 represents the 5 substituents positions 2-6) on phenyl ring J, wherein each R s is independently selected from hydrogen, hydroxy, halo, thiol, -OR 12

-SR

12

SON(R

2

-N(R

2

)(R

12 alkyl, acyl, alkene, alkyne, cyano, nitro, aryl, heteroaryl, cycloalkyl, and heterocycloalkyl; where each R 1 2 and R' 2 is independently selected from hydrogen, alkyl, acyl, heteroalkyl, aryl, heteroaryl, cycloalkyl, and heterocycloalkyl; or two R 5 moieties can optionally join to form a carbocyclic or a heterocyclic ring that is fused to phenyl ring J; q is 0, 1, 2, 3, 4 or 5; wherein when q is greater than 0, each R 6 and R 6 is independently selected from hydrogen, alkyl, aryl, halo, hydroxy, alkoxy, amino and acylamino; when q is greater than 1, two R 6 moieties, together with the carbon atoms to which they are bonded, can join to form a heterocycloalkyl, cycloalkyl or aryl ring; and when q is greater than 1, the R 6 moieties on two adjacent carbon atoms can be nil such that a double bond is formed between the two adjacent carbon atoms, or both the R 6 and R 6 moieties on two adjacent carbon atoms can all be nil such that a triple bond is formed between the two adjacent carbon atoms; Ar is an aryl or heteroaryl ring selected from the group consisting of phenyl, thiophene, furan, oxazole, thiazole, pyrrole and pyridine;

R

7 represents all the substituents on ring Ar, wherein each R 7 is independently selected from hydrogen, halo, -NR 13

R

3 alkyl, acyl, alkene, alkyne, cyano, nitro, aryl, heteroaryl, cycloalkyl, and heterocycloalkyl; where each R 1 3 and R" 1 is independently selected from hydrogen, alkyl, acyl, heteroalkyl, aryl, heteroaryl, cycloalkyl, and heterocycloalkyl; or two R 7 moieties can optionally join to form a carbocyclic or a heterocyclic ring fused to ring Ar; r is 0, 1, 2, 3, 4, 5, 6 or 7; wherein each R 8 and R 8 is independently selected from hydrogen, alkyl, halo, hydroxy, alkoxy and amino; WO 02/26774 PCT/US01/30051 when r is greater than 1, two R 8 moieties, together with the carbon atoms to which they are bonded, can join to form a heterocycloalkyl, cycloalkyl or aryl ring; and when r is greater than 1, the R 8 moieties on two adjacent carbon atoms can be nil such that a double bond is formed between the two adjacent carbon atoms, or both the R 8 and R 8 moieties on two adjacent carbon atoms can all be nil such that a triple bond is formed between the two adjacent carbon atoms; B is selected from -N(R14)C(=NR 15 or =S)NR1 6 R17, -NR 20

R

2 1 cyano a heteroaryl ring eg. thiophene, an alkyl or dialkyl amine, a heteroaryl ring containing at least one ring nitrogen atom and a heterocycloalkyl ring containing at least one ring nitrogen atom, wherein R 1 4

R

15 R 1

R

17

R

20 and R 1 are independently selected from hydrogen, alkyl, alkene, and alkyne; wherein further a combination of two or more of R 4

R

1 5

R

16 and R 17 may optionally combine with the atoms to which they are bonded to form a monocyclic or bicyclic ring; preferred are -N(R 1)C(=NR5)NR16R 7 cyano, N(R1)C(=O)NRR 17 a heteroaryl ring containing at least one ring nitrogen atom and a heterocycloalkyl ring containing at least one ring nitrogen atom. More preferred are N(R1 4 )C(=NRIS)NR1 6

R

17

N(R'

4 )C(=O)NR1 6

R

1 7 cyano, and triazole and imidazole.

s is 0, 1, 2, 3, 4 or 5; wherein when s is greater than 0, each R 9 and R 9 is independently selected from hydrogen, alkyl, aryl, halo, hydroxy, alkoxy, amino and acylamino; when s is greater than 1, two R 9 moieties, together with the carbon atoms to which they are bonded, can join to form a heterocycloalkyl, cycloalkyl or aryl ring; and when s is greater than 1, the R 9 moieties on two adjacent carbon atoms can be nil such that a double bond is formed between the two adjacent carbon atoms, or both the R 9 and R 9 moieties on two adjacent carbon atoms can all be nil such that a triple bond is formed between the two adjacent carbon atoms;

R'

1 is selected from the group consisting of an optionally substituted bicyclic aryl ring and an optionally substituted bicyclic heteroaryl ring; and D is independently selected from hydrogen, fluoro, hydroxy, thiol, acylthio, alkoxy, aryloxy, alkylthio, acyloxy, cyano, amino, acylamino, and wherein R" is selected from the group consisting of hydroxy; alkoxy; amino; alkylamino; NHOR"', where R" 8 is selected from hydrogen and alkyl; -N(R 19

)CH

2

C(O)NH

2 where R' 9 is alkyl; -NHCH 2

CH

2 OH; -N(CH 3

)CH

2

CH

2 0H; and -NHNHC(=Y)NH2, where Y is selected from O, S and NH; and wherein if at least one of Z1, Z 2 or Z 3 is other than -C(O)N(R 3 or -N(R1)CQ0)., then X and D may optionally be linked together via a linking moiety, L, that contains all covalent bonds or covalent bonds and an ionic bond so as to form a cyclic peptide analog; or an optical isomer, diastereomer or enantiomer thereof;, a pharmaceutically-acceptable salt, hydrate, or biohydrolyzable ester, amide or imide thereof.

The present invention also provides a compound having a structure according to Formula R1 0 0 0 V/ 71- -c {C-NH-CH-C-N-H- II (c44)(CR R 6 ?112 P q 3 B R7 7 The invention also relates to pharmaceuti cal compositions comprising the above compo~inds, and to methods of treating disorders mediated by the MC-3 or MC-4 receptor by administering these compounds. Thus, in one aspect, the present invention provides a pharmaceutical composition comprising: a safe and effective amount of a cyclic peptide analog as defined herein a pharmaceutical ly-acceptable excipient.

DETARiLE DESCRIPTION OF TBE RiVENTION L. Deffnitions: "Amino acid" refers t o alanine (Ala; arginine (Arg; asparagie (Asn; aspartic acid.(Asp; cysteine (Cys; glutamic acid (Glu; g] utamine (Gin; glycine (Gly; G), bistidine (is; Hi), isoleucine (Ile; leucine (Leu; lysine (Lys; methionine (Met; phenylalanine (Phe; proline (Pro; serine (Ser; threonine (Thr; tryptophan (Trp; W), tyrosine (Tyr; and valine (Val; The common 3-letter and I-letter abbreviations are indicated parenthetically. Modified amino acids also useful herein are the following (the 3-letter abbreviation for each moiety is noted parenthetically): p-Benzoyl-phenylalanine (Bpa); f3-( l-Naphthil)-alanine (1-Nal); f0-(2-Naphthyl)-alanine (2-Nal); 1-Cyclohexylalanine (Cha), 3 ,4-Dichlorolphenylalanine (3,4-Dep); 4-Fluorophenylalanine 4-Nirophenylalanine (4-Npa); 2-Thienylalanine (Tha); l, 2 ,3, 4 -Tetrahydroisoqinoline-3.carboxylic acid (Tic); 3-Benzothienylalanine (3-Bal); 4-Cyanophenylalanine (4-Ypa); 4 -Iodophenyialanine (4-lpa); S4-Bromophenylalanine (4-Rpa); 4 ,4'-Biphenylalanine (Bip); Ornithine (Orn); Sarcosine (Sar); Pentafluorophenylalanine (Pfp); and P,P-Diphenylalanine (Dip). With respect to moieties depicted on Formula and Formula moieties referred to using a single letter designation are as defined and do not refer to the single letter amino acids corresponding to those letters.

The letter preceding the above three-letter abbreviations, e.g. as in "D-Nal" or S"D-Phe", denotes the D-form of the amino acid. The letter preceding an amino acid threeletter abbreviation denotes the natural L-form of the amino acid. For purposes of this disclosure, N unless otherwise indicated, absence of a or designation indicates that the abbreviation 1 .0 refers to both the D- and L-forms. Where the common single-letter abbreviation is used, O capitalization refers to the L-form and small letter designation refers to the D-form, unless otherwise indicated.

WO 02/26774 PCT/US01/30051 "Ac" refers to acetyl CH3C(=O)-).

"Acylamino" refers to "Acyloxy" refers to "Acylthio" refers to "Alkoxy" is an oxygen radical having a hydrocarbon chain substituent, where the hydrocarbon chain is an alkyl or alkene -O-alkyl or -O-alkene). Preferred alkoxy groups include (for example) methoxy (MeO), ethoxy, propoxy and allyloxy.

"Alkyl" is a saturated hydrocarbon chain having 1 to 15 carbon atoms, preferably 1 to more preferably 1 to 4 carbon atoms. "Alkene" is a hydrocarbon chain having at least one (preferably only one) carbon-carbon double bond and having 2 to 15 carbon atoms, preferably 2 to 10, more preferably 2 to 4 carbon atoms. "Alkyne" is a hydrocarbon chain having at least one (preferably only one) carbon-carbon triple bond and having 2 to 15 carbon atoms, preferably 2 to more preferably 2 to 4 carbon atoms. Alkyl, alkene and alkyne chains (referred to collectively as "hydrocarbon chains") may be straight or branched and may be unsubstituted or substituted. Preferred branched alkyl, alkene and alkyne chains have one or two branches, preferably one branch. Preferred chains are alkyl. Alkyl, alkene and alkyne hydrocarbon chains each may be unsubstituted or substituted with from 1 to 4 substituents; when substituted, preferred chains are mono-, di-, or tri-substituted. Alkyl, alkene and alkyne hydrocarbon chains each may be substituted with halo, hydroxy, aryloxy phenoxy), heteroaryloxy, acyloxy acetoxy), carboxy, aryl phenyl), heteroaryl, cycloalkyl, heterocycloalkyl, spirocycle, amino, amido, acylamino, keto, thioketo, cyano, or any combination thereof. Preferred hydrocarbon groups include methyl ethyl, propyl, isopropyl, butyl, vinyl, allyl and butenyl.

Also, as referred to herein, a "lower" alkyl, alkene or alkyne moiety "lower alkyl") is a chain comprised of 1 to 6, preferably from 1 to 4, carbon atoms in the case of alkyl and 2 to 6, preferably 2 to 4, carbon atoms in the case of alkene and alkyne.

"Alkylthio" is a sulfur radical having a hydrocarbon chain substituent, where the hydrocarbon chain is an alkyl or alkene -S-alkyl or -S-alkene). Preferred alkylthio groups include (for example) methylthio (MeS) and ethylthio.

"Aryl" is an aromatic hydrocarbon ring. Aryl rings are monocyclic or fused bicyclic ring systems. Monocyclic aryl rings contain 6 carbon atoms in the ring. Monocyclic aryl rings are also referred to as phenyl rings. Bicyclic aryl rings contain from about 8 to about 17 carbon atoms, preferably about 9 to about 12 carbon atoms in the ring. Bicyclic aryl rings include ring systems wherein one ring is aryl and the other ring is aryl, cycloalkyl, or heterocycloalkyl.

WO 02/26774 PCT/US01/30051 Preferred bicyclic aryl rings comprise 6- or 7-membered rings fused to or 7-membered rings. Aryl rings may be unsubstituted or substituted with from 1 to 4 substituents on the ring.

Aryl may be substituted with halo, cyano, nitro, hydroxy, carboxy, amino, acylamino, alkyl, heteroalkyl, haloalkyl, phenyl, aryloxy, heteroaryloxy, or any combination thereof. Preferred aryl rings include naphthyl, tolyl, xylyl, and phenyl. The most preferred aryl ring radical is phenyl.

"Aryloxy" is an oxygen radical having an aryl substituent -O-aryl). Preferred aryloxy groups include (for example) phenoxy, naphthyloxy, methoxyphenoxy, and methylenedioxyphenoxy.

As used herein, "basic amino acids" refers to His, Lys, and Arg.

"Bc" refers to butanoyl CH 3

CH

2

CH

2 "Cycloalkyl" is a saturated or unsaturated hydrocarbon ring. Cycloalkyl rings are not aromatic. Cycloalkyl rings are monocyclic, or are fused, spiro, or bridged bicyclic ring systems.

Monocyclic cycloalkyl rings contain from about 3 to about 9 carbon atoms, preferably from 3 to 7 carbon atoms in the ring. Bicyclic cycloalkyl rings contain from 7 to 17 carbon atoms, preferably is from about 7 to about 12 carbon atoms in the ring. Preferred bicyclic cycloalkyl rings comprise 6- or 7-membered rings fused to or 7-membered rings. Cycloalkyl rings may be unsubstituted or substituted with from 1 to 4 substituents on the ring. Cycloalkyl may be substituted with halo, cyano, alkyl, heteroalkyl, haloalkyl, phenyl, keto, hydroxy, carboxy, amino, acylamino, aryloxy, heteroaryloxy, or any combination thereof. Preferred cycloalkyl rings include cyclopropyl, cyclopentyl, and cyclohexyl.

"Fused" refers to cyclic moieties having at least two common ring atoms, the preferred maximum number of fused cycles being three.

"Halo" is fluoro chloro bromo (Br) or iodo "Heteroatom" is a nitrogen, sulfur, or oxygen atom, to which one or more moieties may be connected according to heteroatom valence; in the case of nitrogen, one oxygen atom may be optionally connected to it by a coordinate covalent bond, such as forming an N-oxide. Groups containing more than one heteroatom may contain different heteroatoms.

"Heteroalkyl" is a saturated or unsaturated chain containing carbon and at least one heteroatom, wherein no two heteroatoms are adjacent. Heteroalkyl chains contain from 2 to about 15 member atoms (carbon and heteroatoms) in the chain, preferably 2 to about 10, more preferably 2 to about 5. For example, alkoxy -O-alkyl or -O-heteroalkyl) radicals are included in heteroalkyl. Heteroalkyl chains may be straight or branched. Preferred branched heteroalkyl have one or two branches, preferably one branch. Preferred heteroalkyl are saturated.

WO 02/26774 PCT/US01/30051 Unsaturated heteroalkyl have one or more double bonds (also referred to herein as "heteroalkenyl") and/or one or more triple bonds (also referred to herein as "heteroalkynyl").

Preferred unsaturated heteroalkyl have one or two double bonds or one triple bond, more preferably one double bond. Heteroalkyl chains may be unsubstituted or substituted with from 1 to 4 substituents. Preferred substituted heteroalkyl are mono-, di-, or tri-substituted. Heteroalkyl may be substituted with lower alkyl, halo, hydroxy, aryloxy, heteroaryloxy, acyloxy, carboxy, monocyclic aryl, heteroaryl, cycloalkyl, heterocycloalkyl, spirocycle, amino, acylamino, amido, keto, thioketo, cyano, or any combination thereof.

"Heterocycloalkyl" is a saturated or unsaturated, non-aromatic ring containing carbon and from 1 to about 4 (preferably 1 to 3) heteroatoms in the ring, wherein no two heteroatoms are adjacent in the ring and no carbon in the ring that has a heteroatom attached to it also has a hydroxyl, amino, or thiol radical attached to it. Heterocycloalkyl rings are monocyclic, or are fused, bridged, or spiro bicyclic ring systems. Monocyclic heterocycloalkyl rings contain from about 4 to about 9 member atoms (carbon and heteroatoms), preferably from 5 to 7 member atoms in the ring. Bicyclic heterocycloalkyl rings contain from about 7 to about 17 atoms, preferably from 7 to 12 atoms. Bicyclic heterocycloalkyl rings may be fused, spiro, or bridged ring systems. Preferred bicyclic heterocycloalkyl rings comprise 6- or 7-membered rings fused to or 7-membered rings. Heterocycloalkyl rings may be unsubstituted or substituted with from 1 to 4 substituents on the ring. Heterocycloalkyl may be substituted with halo, cyano, hydroxy, carboxy, keto, thioketo, amino, acylamino, acyl, amido, alkyl, heteroalkyl, haloalkyl, phenyl, phenoxy or any combination thereof. Preferred substituents on heterocycloalkyl include fluoro and alkyl.

"Heteroaryl" is an aromatic ring containing carbon and from 1 to about 4 heteroatoms in the ring. Heteroaryl rings are monocyclic or fused bicyclic ring systems. Monocyclic heteroaryl rings contain from about 5 to about 9 member atoms (carbon and heteroatoms), preferably 5 or 6 member atoms in the ring. Bicyclic heteroaryl rings contain from about 8 to about 17 member atoms, preferably about 8 to about 12 member atoms in the ring. Bicyclic heteroaryl rings include ring systems wherein one ring is heteroaryl and the other ring is aryl, heteroaryl, cycloalkyl, or heterocycloalkyl. Preferred bicyclic heteroaryl ring systems comprise 6- or 7membered rings fused to or 7-membered rings. Heteroaryl rings may be unsubstituted or substituted with from 1 to 4 substituents on the ring. Heteroaryl may be substituted with halo, cyano, nitro, hydroxy, carboxy, amino, acylamino, alkyl, heteroalkyl, haloalkyl, phenyl, aryloxy, WO 02/26774 PCT/US01/30051 heteroaryloxy, or any combination thereof. Preferred heteroaryl rings include thienyl, thiazolo, imidazyl, purinyl, pyrimidyl, pyridyl, and furanyl.

As used herein, "MC-4 agonist" and "MC-3 agonist" refers to a compound with affinity for the MC-4 receptor or MC-3 receptor, respectively, -that results in measurable biological activity in cells, tissues, or organisms which contain the MC-4 or MC-3 receptor. As used herein, an "MC-4/MC-3 agonist" refers to a compound that is both an MC-4 agonist and an MC-3 agonist, as those terms are defined herein. Assays which demonstrate MC-4 and/or MC-3 agonistic activity of compounds are well known in the art. One commonly used assay is the BioTrak TM cAMP direct enzymeimmunoassay (EIA) system from Amersham Pharmacia Biotech (Piscataway, NJ), which quantitates the cAMP response of cells to MC ligands. Another useful assay is the Tropix cAMP ScreenTM, which is available from Tropix. These systems allow the simple quantitation of total cellular cAMP measurement in cells exposed to selective ligands.

Briefly summarized: HEK cells stably transfected with the MC-1, MC-3 or MC-4 receptors are plated into 96 well microtiter plates and grown overnight. Cells are dosed with the appropriate MC ligand for 1 hour and then lysed. A fraction of the lysed cell extract is transferred to the assay plate. The ELISA assay is performed according to kit instructions. Each plate contains a series of cAMP standards for calculating a standard curve, as well as a full MC agonist as a positive control for each MC receptor. cAMP activity is calculated as a of the maximum cAMP activity of the full MC agonist control.

As used herein, "MC-4 antagonist" and "MC-3 antagonist" refer to compounds with affinity for the MC-4 receptor or MC-3 receptor, respectively, and blocks stimulation by a known MC agonist. As used herein, an "MC-4/MC-3 antagonist" refers to a compound that is both an MC-4 antagonist and an MC-3 antagonist, as those terms are defined herein. Assays that demonstrate compounds with MC-4 and/or MC-3 antagonism are well known in the art. One particularly useful assay is competitive binding with the use of Europium-labeled NDP-MSH.

Briefly summarized: HEK cells stably transfected with the MC-1, MC-3, MC-4 or receptors are plated into 96-well microtiter plates and grown overnight. Cells are dosed with the appropriate MC ligand in the presence of europilated-NDP-MSH for 60 min., cells are washed several times, enhancement solution is added and fluorescence is measured. IC5o and Ki values can be calculated at each receptor for each MC ligand using standard graphing programs such as GraphPad PrismTM, (GraphPad Software Inc., San Diego, CA).

As used herein, "MC-3 receptor" and "MC-4 receptor" mean the known MC-3 and MC-4 receptors, their splice variants, and undescribed receptors. MC-3 receptors are described by WO 02/26774 PCT/US01/30051 Gantz et al., supra (human MC-3); Desarnaud et al., supra (mouse MC-3) and L. Reyfuss et al., "Identification of a Receptor for Gamma Melanotropin and Other Proopiomelanocortin Peptides in the Hypothalamus and Limbic System., Proc. Natl. Acad. Sci. USA, vol. 90, pp. 8856-8860 (1993) (rat MC-3). MC-4 receptors are described by Gantz et al., supra (human MC-4), J.D.

Alvaro et al., "Morphine Down-Regulates Melanocortin-4 Receptor Expression in Brain Regions that Mediate Opiate Addiction", Mol-Pharmnacol. Sep, vol. 50(3), pp. 583-91 (1996) (rat MC-4) and Takeuchi, S. and Takahashi, "Melanocortin Receptor Genes in the Chicken--Tissue Distributions", Gen-Comp-Endocrinol., vol. 112(2), pp 220-31 (Nov. 1998) (chicken MC-4).

As used herein, "measurable" means the biologic effect is both reproducible and significantly different from the baseline variability of the assay.

A "pharmaceutically-acceptable salt" is a cationic counterion formed at any acidic (carboxylic acid) group, or an anionic counterion formed at any basic amino) group. Many such salts are known in the art, as described in World Patent Publication 87/05297, Johnston et al., published September 11, 1987 incorporated by reference herein. Preferred cationic salts include the alkali metal salts (such as sodium and potassium), and alkaline earth metal salts (such as magnesium and calcium) and organic salts. Preferred anionic salts include the halides (such as chloride salts), sulfonates, carboxylates, phosphates, trifluoroacetate (TFA) and the like. Clearly contemplated in such salts are addition salts that may provide an optical center where once there is none. For example, a chiral tartrate salt may be prepared from the compounds of the invention, and this definition includes such chiral salts.

Such salts are well understood by the skilled artisan, and the skilled artisan is able to prepare any number of salts given the knowledge in the art. Furthermore, it is recognized that the skilled artisan may prefer one salt over another for reasons of solubility, stability, formulation ease and the like. Determination and optimization of such salts is within the purview of the skilled artisan's practice.

As used herein, "selective" means having an activation preference for a specific receptor over other receptors which can be quantified based upon whole cell, tissue, or organism assays which demonstrate receptor activity, such as the cAMP enzyme immunoassay (EIA) system discussed above. A compound's selectivity is determined from a comparison of its EC 5 0 values at the relevant receptors being referenced. As used herein, unless indicated, use of the term "selective over the other MC receptors" means selective with respect to the other melanocortin receptors, including the MC-1, MC-2 and MC-5 receptors. For example, a compound having an

EC

5 s of 8 nM at the MC-4 receptor and an EC5o of 2 80 nM at the MC-1, MC-2 and WO 02/26774 PCT/US01/30051 receptors has a selectivity ratio for the MC-4 receptor over the other MC receptors of at least 1:10. Additionally, it will be recognized that selectivity may also refer to one of the MC-1, MC-2 or MC-5 receptors individually. For example, a compound having an ECso of 8 nM at the MC-4 receptor and an ECso of 80 nM at the MC-1 receptor has a selectivity ratio for the MC-4 receptor over the MC-1 receptor of 1:10. Such a compound is selective over the MC-1 receptor, regardless of its ECs 0 value for MC-2 or MC-5. Selectivity is described in more detail below and may be determined by using, for example, the software Prism v 2.0 which is available from GraphPad, Inc.

A "solvate" is a complex formed by the combination of a solute a MC-4/MC-3 receptor ligand of the present invention) and a solvent water). See J. Honig et al., The Van Nostrand Chemist's Dictionary, p. 650 (1953). Pharmaceutically-acceptable solvents used according to this invention include those that do not interfere with the biological activity of the compound water, ethanol, ethers, acetic acid, N,N-dimethylformamide and others known or readily determined by the skilled artisan).

"Spirocycle" is an alkyl or heteroalkyl diradical substituent of an alkyl or heteroalkyl, wherein said diradical substituent is attached geminally and wherein said diradical substituent forms a ring, said ring containing about 4 to about 8 member atoms (carbon or heteroatoms), preferably 5 or 6 member atoms.

II. The Compounds The compounds of the present invention are MC-4 and/or MC-3 receptor ligands having a structure according to Formula R2 R2 R 2

R

2 I I 2 I 3 I X-C--Z -C Z-C--Z-C-D (C o R 6 R I) (CR 8

R

8

R

SB Ri 2 6 Ar 3 45 R R wherein R 2

R

4

R

4

R

5

R

6

R

6 7

R

8

R

8

R

9

R

9

R

10 Ar, Z 1

Z

2 Z, X, B, D,p, q, r and s are as described in the Disclosure of the Invention section above.

In addition to the compounds described by Formula it is envisioned that the core peptidic residues can be pegylated to provide enhanced therapeutic benefits such as, for example, WO 02/26774 PCT/US01/30051 increased efficacy by extending half-life in vivo. Peptide pegylation methods are well known in the literature. For example, pegylation of peptides is described in the following references, the disclosure of each of which is incorporated herein by reference: Lu, Y.A. et al., "Pegylated peptides. II. Solid-phase synthesis of amino-, carboxy- and side-chain pegylated peptides", Int. J.

Pept. Protein Res., Vol. 43(2), pp. 127-38 (1994); Lu, Y.A. et al., "Pegylated peptides. I. Solidphase synthesis of N alpha-pegylated peptides using Fmoc strategy", Pept. Res., Vol. pp.

140-6 (1993); Felix, A.M. et al., "Pegylated peptides. IV. Enhanced biological activity of sitedirected pegylated GRF analogs.", Int. J. Pept. Protein Res., Vol. 46(3-4), pp. 253-64 (1995); Gaertner, H.F. et al., "Site-specific attachment of functionalized poly(ethylene glycol) to the amino terminus of proteins", Bioconjug Chem., Vol. pp. 38-44 (1996); Tsutsumi, Y. et al., "PEGylation of interleukin-6 effectively increases its thrombopoietic potency", Thromb Haemost, Vol. 77(1), pp. 168-73 (1997); Francis, G.E. et al., "PEGylation of cytokines and other therapeutic proteins and peptides: the importance of biological optimisation of coupling techniques", Int. J. Hematol., Vol. 68(1), pp. 1-18 (1998); Roberts, M.J. et al., "Attachment of degradable poly(ethylene glycol) to proteins has the potential to increase therapeutic efficacy", J.

Pharm. Sci., Vol 87(11), pp. 1440-45 (1998); and Tan, Y. et al., "Polyethylene glycol conjugation of recombinant methioninase for cancer therapy", Protein Expr. Purif, Vol. 12(1), pp. 45-52 (1998). The compounds of Formula can be pegylated directly, or a "linker arm" may be added to the compounds to facilitate pegylation.

With reference to Formula the following is a non-limiting list of preferred substituents: X is selected from hydrogen, fluoro, aryloxy, acyloxy, OR 1 SR', -NRR 1 and -CHR'R'.

Preferred are hydrogen (when D is not also hydrogen), -NR R' and -CHR'R'. More preferred are -NR1R' and -CHRR 1 Still more preferred is -NR 1

R'.

R

1 and R' are independently selected from the group consisting of hydrogen, alkyl and acyl. Preferred is where R' is hydrogen or alkyl and R" is acyl.

R

2 is independently selected from the group consisting of hydrogen, alkyl halo and heteroalkyl. Preferred is hydrogen. Alternatively, two consecutive R 2 moieties, or consecutive

R

2 and R 3 moieties, may join to form a 3 to 8 membered carbocyclic or heterocyclic ring. In another alternative, the Rz bonded to the carbon atom that is bonded to X and Z' and an R 5 moiety can optionally join to form a carbocyclic or heterocyclic ring that is fused to phenyl ring J. In another alternative, the R 2 bonded to the carbon atom that is bonded to ring Ar can join with an

R

7 to form a ring fused to ring Ar. In another alternative, the R 2 bonded to the carbon atom that is WO 02/26774 WO 0226774PCT/USOI/30051 bonded to Z 2 and Z 3 can optionally join with R 8 to form a carbocyclic or heterocyclic ring. In still another alternative, the R2 bonded to the carbon atom that is bonded to f 3 and D can optionally join with R 10 to form n carbocyclic or heterocyclic ring. With respect to the foregoing, preferred is where R2 does not form a ring with another R7, and where W 2 does not form a ring with R 3, R7 or R8. More preferred is where R7 also does not form a ring with R1 0 Preferably rings formed between R2 and another moiety will have 5 to 8 ring atoms.

Each of Z 1

Z

2 and Z 3 is independently selected from -OC(R 3

)(R

3

-C(R

3

)(R

3 S(O)aC(R 3

)(R

3 where a is 0, 1 or 2; -C(R 3 3 where b is 0, 1 or 2; -N(R 3 )C(R 3 3

C(RS)(R

3 a)N(R 3

-C(O)N(R

3

-N(R

3

_C(O)C(R

3

)(R

3 a _C(R 3 3 C(R 3 3 a)C(R 3

_C(R

3

_SO

2

N(R

3 _N(Rld)SO 2

C(R

3

-P(=O)(OR

3 f)C(R 3

)(R

3 3

-P(=O)(OR

3 f)N(R 3 P(=O)(0R 3 0 -O-P(=O)(0R 3 a cycloalkyl having from 3 to 8 ring atoms and a heterocycloalkyl having from 4 to 8 ring atoms. Preferred are -OC(Rs)(RWa)_; _C(R 3 S(O)aC(R 3

)(R

3 where a is 2; -C(R 3 3 where b' is 2; -N(R 3 e)C(R 3(Rla)_; (R 3 3 a -N(R -C(R 3 )(Rla)C(RWb)(Rlc)_; _C(R 3

SO

2 N(R 3

_N(R

3 d)SO 2 and -P(=O)(ORM)C(R 3 3 More preferred are -OC(R 3

-C(R

3

C(R

3 )(RWa)N(Re~ -C(O)N(Rd~ I (R 3

)(R

3 a)C(Rb)(R 3

-C(R

3 3

_SO

2 N(Rld)_ and _p(=O)(OR 3 f)C(R 3 Most preferred are -C(R 3 3

-C(O)N(R

3 and _C(R 3

)(R

3 a)C(R 3 b)(R 3 In another aspect, preferred compounds are those where at least one of Z or Z is other than More preferred are compounds where at least two of f 2 or Z 3 aeother than -C(O)N(RW Also more preferred are compounds where at least one of Z 1

Z

2 or Z 3 is other than and (ii)(a) X is other than -NR 1 R" where is acyl and/or D is other than -C(O)R' 1 Each of RI, R 3 R Rb and RWO, when present, is independently selected from hydrogen, hydroxy, alkoxy, aryloxy, acyloxy, thiol, alkylthio, acylthio, arylthio, amino, alkylamino, acylamino, and alkyl. Preferred are hydrogen, hydroxy, alkoxy, aryloxy and alkyl. Most preferred is where each of R"a R Ib and R 3 C is hydrogen.

R 3 dI when present, is selected from hydrogen, alkyl and aryl. Preferred is where R 3 d is selected from hydrogen and alkyl.

when present, is selected from hydrogen, alkyl, aryl and acyl. Preferred is where R 3 is selected from hydrogen and alkyl.

R

3 f is selected from hydrogen and alkyl. When R 3 f is alkyl, preferred are branched alkyl, preferably isopropyl.

WO 02/26774 PCT/US01/30051 p is 0, 1, 2, 3, 4 or 5. Preferablyp is 1 or 2, more preferably 1.

When p is greater than 0, each R 4 and R 4 is independently selected from hydrogen, alkyl, aryl, halo (preferably fluoro), hydroxy, alkoxy, amino and acylamino. When p is greater than 1, two R 4 moieties, together with the carbon atoms to which they are bonded, may join to form a heterocycloalkyl, cycloalkyl or aryl ring. When p is greater than 1, the R 4 moieties on two adjacent carbon atoms can both be nil such that a double bond is formed between the two adjacent carbon atoms, or both the R 4 and R 4 moieties on two adjacent carbon atoms can all be nil such that a triple bond is formed between the two adjacent carbon atoms. Preferably each R 4 when present, is hydrogen and each R 4 when present, is hydrogen or alkyl. Most preferably there is no unsaturation in the chain linking ring J to the X-containing carbon atom of Formula

R

5 represents the 5 substituents positions 2-6) on phenyl ring J, wherein each R 5 is independently selected from hydrogen, hydroxy, halo, thiol, -OR 1 2

-SR

12

-SO

2 N(R12)(R 2 N(R12)(RI' 2 alkyl, acyl, alkene, alkyne, cyano, nitro, aryl, heteroaryl, cycloalkyl, and heterocycloalkyl. Each R 12 and R 12 is independently selected from hydrogen, alkyl, acyl, heteroalkyl, aryl, heteroaryl, cycloalkyl, and heterocycloalkyl; or two R 5 moieties can optionally join to form a carbocyclic or a heterocyclic ring that is fused to phenyl ring J.

Preferred R 5 moieties are hydrogen, hydroxy, halo, thiol, -OR 12 where R 12 is lower alkyl or acyl, -SR 12 where R 12 is lower alkyl or acyl, -SO 2 N(R12)(R' 2 -N(R12)(R 12 alkyl, cyano, nitro, aryl, heteroaryl, cycloalkyl, and heterocycloalkyl. More preferred R 5 moieties are hydrogen, hydroxy, halo, thiol, -SO 2

N(R')(R

1 where R 1 2 and R 2 both are hydrogen, -N(R' 2

)(R

1 2 where

R'

2 and R 2 each are hydrogen or alkyl. Still more preferred R 5 moieties are hydrogen, hydroxy, chloro, fluoro, -N(Rl)(R 2 where R 12 and R' 2 each are hydrogen or alkyl. Most preferred R moieties are hydrogen, hydroxy, chloro, fluoro and nitro.

With respect to ring J, preferred is where four of the R 5 moieties are hydrogen. Also preferred is where the 4-position is other than hydrogen. Most preferred is where the 4-position is other than hydrogen and the remaining 4 substituents are hydrogen.

q is 0, 1, 2, 3, 4 or 5. Preferably, q is 0, 1 or 2, more preferably q is 1.

When q is greater than 0, each R 6 and R 6 is independently selected from hydrogen, alkyl, aryl, halo (preferably fluoro), hydroxy, alkoxy, amino and acylamino. When q is greater than 1, two R 6 moieties, together with the carbon atoms to which they are bonded, can join to form a heterocycloalkyl, cycloalkyl or aryl ring. When q is greater than 1, the R 6 moieties on two adjacent carbon atoms can be nil such that a double bond is formed between the two adjacent WO 02/26774 PCT/US01/30051 carbon atoms, or both the R 6 and R 6 moieties on two adjacent carbon atoms can all be nil such that a triple bond is formed between the two adjacent carbon atoms. Preferably each R 6 when present, is hydrogen and each R when present, is hydrogen or alkyl. Most preferably there is no unsaturation in the chain linking the ring Ar to the carbon atom of Formula that is bonded to

Z

1 and Z 2 Ar is an aryl or heteroaryl ring selected from the group consisting of phenyl, thiophene, furan, oxazole, thiazole, pyrrole and pyridine. Ar is preferably phenyl, thiophene or furan. Ar is most preferably phenyl.

R

7 represents the substituents on the Ar ring, wherein each R 7 is independently selected from hydrogen; halo; -NR" 3

R

1 where R 13 and R 1 3 each are hydrogen or alkyl; alkyl; acyl; alkene; alkyne; cyano; nitro; aryl; heteroaryl; cycloalkyl and heterocycloalkyl. Optionally, two R 7 moieties can join to form a carbocyclic or a heterocyclic ring fused to ring Ar. When Ar is phenyl, preferred is where four of the five R 7 moieties are hydrogen or all five of the R 7 moieties are hydrogen. Also preferred is where two R 7 moieties are selected from fluoro, chloro, cyano, bromo, iodo, nitro, alkoxy and alkyl; or two R 7 moieties join to form a carbocyclic or a heterocyclic ring fused to the phenyl ring. More preferred is where the 4-position of the phenyl ring is hydrogen, fluoro, chloro, cyano; bromo, iodo, nitro and alkyl and the remaining four positions are hydrogen. Most preferred is where the 4-position of the phenyl ring is hydrogen or fluoro and the remaining four substituents are hydrogen.

When Z 1 and Z 2 are both -C(O)N(R 3 peferred are compounds where the carbon atom that is bonded to Z 1 and Z 2 is assigned an R configuration according to Cahn-Ingold-Prelog rules of nomenclature.

r is 0, 1, 2, 3, 4, 5, 6 or 7. Preferred r is 2, 3, 4 or 5. More preferred r is 2, 3 or 5. Most preferred r is 3.

Each R 8 and R s is independently selected from hydrogen, alkyl, halo (preferably fluoro),.

hydroxy, alkoxy and amino. Optionally, when r is greater than 1, two R 8 moieties, together with the carbon atoms to which they are bonded, join to form a heterocycloalkyl, cycloalkyl or aryl ring. Preferably, each R 8 and R 8 is independently selected from hydrogen and alkyl. Most preferably, each R 8 and R 8 is hydrogen. Optionally, when r is greater than 1, the R 8 moieties on two adjacent carbon atoms can be nil such that a double bond is formed between the two adjacent carbon atoms, or both the R 8 and R 8 moieties on two adjacent carbon atoms can all be nil such that a triple bond is formed between the two adjacent carbon atoms.

WO 02/26774 PCT/US01/30051 B is selected from -N(R 4

)C(=NR

15 or =S)NR16R 17

-NRR

21 cyano a heteroaryl ring eg. thiophene, an alkyl or dialkyl amine, a heteroaryl ring containing at least one ring nitrogen atom and a heterocycloalkyl ring containing at least one ring nitrogen atom.

Preferred are -N(R 1 a)C(=NR 1 5

)NR

16

R

17 a heteroaryl ring containing at least one ring nitrogen atom and a heterocycloalkyl ring containing at least one ring nitrogen atom. More preferred are N(Ri )C(=NR 5 )NR16R17 cyano, N(R1 4

)C(=O)NR

16

R

17 a heteroaryl ring containing at least one ring nitrogen atom and a heterocycloalkyl ring containing at least one ring nitrogen atom. More preferred are N(R14)C(=NR NR1R 17

N(R

14

)C(=O)NRR

1 7 cyano, and triazole and imidazole..

R

1 4 and R 15 are independently selected from hydrogen, alkyl, alkene, and alkyne.

Preferred are hydrogen and alkyl. R 1 6 and R 1 7 are independently selected from hydrogen, alkyl, alkene, and alkyne. Preferred are hydrogen and alkyl. R 20 and R 21 are independently selected from hydrogen, alkyl, alkene, and alkyne. Preferred are hydrogen and alkyl.

Alternatively, a combination of two or more of R 14

R

15

R"

1 and R17 combine to form a monocyclic or bicyclic ring. For example, R 1 4 and R 15 together with the atoms to which they are bonded, can join to form a heterocycloalkyl or a heteroaryl. Also, R 1 4 and R' 6 together with the atoms to which they are bonded, can join to form a heterocycloalkyl or a heteroaryl. Also, R' and R together with the atoms to which they are bonded, can join to form a heterocycloalkyl or a heteroaryl. Also, R 16 and R 17 can optionally join to form a heteroaryl or heterocycloalkyl ring.

Preferred is where R 15 and R 16 join to form a ring.

s is 0, 1, 2, 3, 4 or 5. Preferably s is 1 or 2, more preferably 1.

When s is greater than 0, each R 9 and R 9 is independently selected from hydrogen, alkyl, aryl, halo (preferably fluoro), hydroxy, alkoxy, amino and acylamino. Preferably each R 9 when present, is hydrogen and each R 9 when present, is hydrogen or alkyl. Optionally, when s is greater than 1, two R 9 moieties, together with the carbon atoms to which they are bonded, join to form a heterocycloalkyl, cycloalkyl or aryl ring. Also, when s is greater than 1, the R 9 moieties on two adjacent carbon atoms can be nil such that a double bond is formed between the two adjacent carbon atoms. Also, when s is greater than 1 both the R 9 and R 9 moieties on two adjacent carbon atoms can all be nil such that a triple bond is formed between the two adjacent carbon atoms. Most preferably there is no unsaturation in the chain linking R 1 0 to the Dcontaining carbon atom of Formula

R

1 is selected from the group consisting of an optionally substituted bicyclic aryl ring and an optionally substituted bicyclic heteroaryl ring. Preferred bicyclic aryl rings include 1- WO 02/26774 WO 0226774PCT/USOI/30051 naphihyl, 2-naplithyl, indan, 1H-indene, benzocylcobutane and benzocylcobutenie. Preferred bicyclic heteroaryl rings include indole, indoline, pyrindine, clihydropyrindine, octahydropyrindine, beuzothiophene, henzofuran, benzimidozole, benzopyran, quinoline, quinolone and isoquinoline. More preferred is where R1 0 is 1-naplithyl, 2-naphthyl, indole, indan, 1H-indene, benzothiophene, beuzofuran and benzopyran. Most preferred is where R 10 is 1naplithyl, 2-naphthyl or indole (particularly 3-indole).

D is selected from hydrogen, fluoro, hydroxy, thiol, alkoxy, aryloxy, alkyithio, acyloxy, cyano, amino, acylamino, -C(C)R' 1 and -C(S)R 1 Preferred are fluoro, hydroxy, thiol, alkoxy, aryloxy, alkylthio, acyloxy, cyano, amiino, acylamino, -C(O)R 1 and -C(S)R" 1 More preferred are alkoxy, cyano, amino, acylamnino, and Still more preferred are -C(O)R 1 and

C(S)R

11 Most preferred is

R

11 is selected from the group consisting of amino; alkylanmino; -NI{0R 1 8, where R' 8 is selected from hydrogen and alkyl; -N(R' 9

)CH

2 C(O)N1 2 where R1 9 is alkyl. (preferably lower alkyl); -NHCH.2CH 2 OH; -N(CH 3

)CH

2

CH

2 OH; and -NHM{C(=Y)NH 2 where Y is selected from 0, S and Nil. Preferred R' 1 are amino; alkylamnino; -N110R 1 8, where R1 8 is selected from hydrogen and alkyl (preferably hydrogen); -N(R' 9

)CH

2

C(O)NH

2 where R19 is alkyl (preferably lower alkyl); -NIICH 2

CH

2 OH; and -N(CH 3

)CH

2

CH

2 OH. More preferred R 11 are amino; alkylamino; -NT{0R 1 where R's is selected from hydrogen and alkyl (preferably hydrogen); and

-N(R'

9

)CH

2 C(0)NH 2 where R' 9 is alkyl. Most preferred are amino and alkylamino.

As is indicated with respect to Formula if at least one of Z 2 or f 3 is other than or -N(R 3 then X and D may optionally be linked together via a linking moiety, L, that contains all covalent bonds or covalent bonds and an ionic bond so as to form a cyclic peptide analog. Such cyclic peptides have a structure according to the following Formula

L

R R R R I 1 1 2 3 Xv-C Z C- Z C Z-C-D (cR R) (CRV 6 (CR R 81 (CR R?) B R j2 R 5 R7 0I0 WO 02/26774 PCT/US01/30051 With respect to cyclic compounds containing linking moiety L, the bridge connecting X and D can be in the form of covalent bond linkages or alternatively can include a salt bridge resulting from the formation of ionic bonds. The linking moiety can be wholly peptidic in nature containing amino acids only), non-peptidic containing no amino acids) in nature, or it can include both peptidic and non-peptidic moieties introduced using well-known chemistry. The linking moiety can comprise aliphatic residues, aromatic residues or heteroaromatic residues, or any combination thereof. In one embodiment, the linking moiety will preferably comprise long chain omega-amino acids in which amino and carboxyl groups are separated by from about 4 to about 24 methylene groups or a combination of said omega-amino acids and aminobenzoic acids.

In another embodiment, which is a preferred embodiment, the linking moiety will contain all covalent bonds, such as amide bonds. For example, the linking moiety can comprise an amide formed through the chemical coupling of a side-chain amino group of amino acids such as Lys or Orn, and a side-chain carboxyl group of the amino acid residue such as Asp or Glu.

Alternatively, the linking moiety can comprise an amide formed between the amino and carboxylate groups attached to the a-carbon of the bridging moiety amino acids. (Hereafter referred to as the "a-amino" moiety of an amino acid or the "a-carboxyl" moiety of an amino acid.) In another alternative, the linking moiety can comprise an amide formed between any combination of the side-chain amino group or side-chain carboxyl group and the a-amino and the a-carboxyl moieties. The linking residues may be amine- or carboxyl-containing structures other than natural amino acids, including, 6-aminohexanoic acid as an amine-containing residue and succinic acid as a carboxyl-containing residue. Furthermore, the invention allows for linking using other types of chemical functionalities. In this case, these linking residues may contain a variety of groups and substituents, including aliphatic, heteroalkyl, aromatic and heterocyclic moieties. When covalently linked, the linking moiety can include but is not limited to amide, ester, ether, thioether, aminoalkyl, aminoaryl, alkyl, other heteroalkyl, alkene, alkyne, heterocycloalkyl, aryl, and heteroaryl. Preferably, the linking moiety can include ether, aminoalkyl, aminoaryl, alkyl, other heteroallky, alkene, alkyne, heterocycloalkyl, aryl, and heteroaryl. More preferably, the linking moiety can include ether, aminoalkyl, alkyl, alkene, and alkyne. When L contains only covalent bonds, preferred are compounds having from about 12 to about 32 ring atoms, more preferred are compounds having from about 22 to about 28 ring atoms.

The linking moiety can alternatively include an ionic bond/association that favors a cyclic structure. This "ionic" bridge is comprised of salt-forming basic and acid functionalities.

For example, the link can comprise an ionic bond formed between the side-chain amino group of WO 02/26774 PCT/US01/30051 amino acids such as Lys or Orn, and the side-chain carboxyl group of the amino acid residue such as Asp or Glu. Alternatively, the linking moiety can comprise an ionic bond formed between the amino and carboxylate groups attached to the a-carbon of the linking moiety amino acids. In still another alternative, the linking moiety may comprise an amide formed between any combination of the side-chain amino group or side-chain carboxyl and the a-amino and the a-carboxyl moieties. When L contains an ionic bond, the ring formed will preferably contain from about 22 to about 28 ring atoms.

It will be recognized that any free peptidic ca-carboxy and a-amino groups amino acid cx-carboxy and a-amino groups) not involved in formation of the ring can optionally be in the form of a carboxyamide or an acylamino moiety, respectively. The most preferred Lcontaining compounds are analogs wherein X and D form covalently bonded cyclic structures.

With respect to the present compounds in general, while alkyl, heteroalkyl, cycloalkyl, and heterocycloalkyl groups may be substituted with hydroxy, amino, and amido groups as stated above, the following are not envisioned in the invention: 1. Enols (OH attached to a carbon bearing a double bond).

2. Amino groups attached to a carbon bearing a double bond (except for vinylogous amides).

3. More than one hydroxy, amino, or amido attached to a single carbon (except where two nitrogen atoms are attached to a single carbon atom and all three atoms are member atoms within a heterocycloalkyl ring).

4. Hydroxy, amino, or amido attached to an sp 3 -hybridized carbon that also has a heteroatom attached to it.

Hydroxy, amino, or amido attached to a carbon that also has a halogen attached to it.

A preferred subclass of compounds of Formula wherein there is no linking moiety L to form a macrocyclic ring are compounds having a structure of Formula as follows: 1

R

2 O O O I 11 II II N -c -Z-CH-C-NH-CH-C-NH-CH-C-R 1

(R

4

R

4

(C

6 R 6

CH

2

RIO

1 2' 1 6' R R(A)

R

7

(A)

WO 02/26774 PCT/US01/30051 In this preferred subgenus of compounds, the moieties R 1

R

4 R R, R6', R, B, R 10 and R 1 are as defined with respect to Formula With reference to Formula the compounds of Formula are those where ring J of Formula is a phenyl ring wherein all of positions 2, 3, 5 and 6 are hydrogen, such that the ring is only substituted at the 4-position with the R 5 moiety, which is as defined with respect to Formula Similar to the description of Formula the R 5 ring moiety and the R 2 substituent can optionally join to form a ring fused to the depicted phenyl ring. In such an embodiment, the fused ring may join the phenyl ring at a position other than the 4-position. In Fomula ring Ar of Formula is a phenyl ring where all of positions 5' and 6' are hydrogen and position 4' is R 7 which is as defined above. In this regard, preferred are where R 7 is selected from hydrogen and fluoro.

With respect to Formula p and q are independently 1 or 2, preferably q is 1. Also preferred are where R 4

R

4

R

6 and R 6 are all hydrogen. Also preferred are compounds where B is -N(R4)C(=NR15)NR R 7 or-NR R21.

A preferred sub-class of compounds of Formula (II) are compounds having a structure according to Formula as follows:

L

-X-CH- Z CH -Z-CH-Z3-CH-D- (CR CR 6

R

6 (CRR )3 CRR p SB R 1 4 5 3' 7

(B)

where X, Z 2

Z

3 D, R 4

R

4

R

5

R

6

R

6

R

7

R

8 B, R 9

R

9 and R 10 are as defined above and p is 1 or 2. Preferred are compounds where R 6 and R 6 are both hydrogen. Also preferred are compounds where B is -N(R4)C(=NRNR R 17 or -NR2R21. Also preferred is where R 8

R

8

R

9 and R 9 are all hydrogen. Preferably R 7 is selected from hydrogen and fluoro. Preferred are N(R 4 )C(=NR )NR 16

R

1 7 cyano, N(R1)C(=O)NR 1 6

R

17 a heteroaryl ring containing at least one ring nitrogen atom and a heterocycloalkyl ring containing at least one ring nitrogen atom. More preferred are N(R )C(=NR )NRR 7

N(R

1 4 )C(=O)NR6R 1 7 cyano, and triazole and imidazole.

The following is a non-limiting list of preferred compounds of Formula With respect to the compounds depicted with chemical structures, included are both "linear" compounds WO 02/26774 WO 0226774PCT/USOI/30051 those where linking moiety L is not present to provide a macrocyclic molecule) and macrocyclic compounds of Formula (11).

The following list uses the one- and/or three-letter amino acid abbreviations discussed above.

YfRW-NH 2 Ac-YfRW-NTI 2 Y(2-Nal)RW-NH 2 Ac-YfRW Y(l-Nal)RW-NHz Ac-Y(D-1-Nal)RW-NH- 2 Ac-aIIDYfRWK] -Nil 2 Ac-a[DY(D-Phe(4-C1))RWK] -NH 2 Ac-Y(2-Nal)RW-NH 2 Ac-IIEYfRWGK] -Nil 2 Ac-Y(D-2-Nal)RW-NH 2 Ac-YFRW-N11 2 Ac-Y(2-Nal)RW Ac-Y(D-Phe(4-F))RW-N1 2 Ac-(Phe(4-F))fRW-NHCH 3 Ac-(Phe(4-Cl))fRW-NH 2 Ac-(Phe(4-Cl))fRW-NHCH 3 Ac-YfKW-N11 2 Ac-YtTK(2-Nal)-N11 2 Ac-YfK(2-Nal)-N{CH 3 Ac-YfR(2-Nal)-NIICH 3 Ac-YfR(2-Nal) -NH 2 Ac-YfR( 1-Nal) -NHz (des- NH 2 Tyr)YfR(2-Nal)-NHCH 3 Ac-TICfRW-NHCH 3 Ac-a[EYfRWGK]-NH 2 2 ))fRW-N1 2 Ac--(Phe(4-Cl))(D-Phe(4-F))RW-NH 2 Ac--(Phe(4-Cl))(D-Phe(4-F))RW- NHCH 3 Bc-YfRW(Sar)-NH 2 Ac-(Phe(4-Cl))fR(2-Nal)-NHCH 3 Bc-(Phe(4-Cl))(fRW(Sar)-NTI 2 Ac-YfHW-NH 2 Ac-Yf(homo-HiEs)W-NH 2 Ac-(Phe(4-Cl))XPhe(4-F)R(2-Nal)-NHCH 3 Ac-Y(Phe(4-F)R(2-Nal)-NHCH3 Ac-FfRW-NH 2 Ac-FfR(2-Nal)-NHCH 3 Ac-yfRW-NH 2 yfRW-NH 2 Ac-YyRW-N11 2 Ac-Y(D-Phe(4-I))KW-NH 2 Ac-Y(D-Phe(4-I))HW- N11 2 Ac-Y(D-Phe(4-I))RW-NH 2 Ac-WfRW-NH 2 Ac-YfR(Trp(5-F)-NH 2 Ac-Y(D-Phe(4Br))RW-NH 2 Ac-(D-Phe(3-OH))fRW-NH 2 Ac-F(D-Phe(4-I))KW]I-Nil 2 Ac-YfR(Trp(5-OMe)-NH 2 2 Ac-YfR(Trp(5-Mc)-NH 2 Ac-a[DYfR(Trp(6-F)CK]-N1 2 Ac-YfR(Tip( 1-Me)-NH 2 Ac-a[DYfR(Trp(4-F)GK]-NH 2 Ac-YfR(Trp(6-Br) -NH 2 Ac-aIIDYfR(Trp(7-Me)GKI-N1 2 Ac-YfR(Trp(5-OH))-N1 2 Ac-YfR(Trp(6-OH)) -NH 2 Ac-YfR(Trp(6-C))-N1 2 WO 02/26774 WO 0226774PCT/USO1/30051 Ac-(Tyr(Me))fRWG-NH 2 Ac-(Tyr(3-NH 2 ))fRW-NH 2 Ac-(Tyr(3-MeO))fR-N{ 2 HN

NH

H2 NNH 0 H 0

NH

0' CH 2 O0 NH O NH

HN,

H

2 N N 0 0 o NH 0GH 0 CHN1 0

~'NH

HN

NH c Ac-(Tyr(CH 2 Ph))fRW-NH 2 Ac(Tyr(3-C))fRW-N1T 2 Ac-Y(D-Phe(5-F))RW-N{ 2 HN

NHN

H

2 N lkN0 0 H H 0 'N0 (CH 2 10 7 "0

NH

SOH

HN

NHN

H

2 N N H 0 O NH 00H 3

NH

SOH

HN NH'0

H

2 N N N 0 H -Hj WO 02/26774 WO 0226774PCT/USO1/30051 HNHN 0 ly0 N0 H 2 9 N N 0 NH NH- 0 HN C 290 H H N N H NH OH 7 OH HN H/ NH 0 'HN N

H

2 N N N 2 N0 H H H 2 0/H H4 Kji. \8NH '7

NNH

O'lNN 0 'N 'N C~ H -~OH H2N 0 HN N 0 NH N H N o CH1 OH WO 02/26774 WO 0226774PCT/USO1/30051

NH

H

2 N

N

H

0

H

2 N N 0 0 ol NH

HN

(CA

I WO 02/26774 WO 0226774PCT/USO1/30051 NH HNH, OiPr P HN (CiPr o H-N HN H NH HN

NHN

HO HO H NH 9 NH HN O~ H O o H-N 0 H-N NH HN NH HO cl H KNH2 jNH2 I NH HN OH HC0 O H-N

H-N

NH NH HO CI WO 02/26774 PCT/US01/30051 OHN NH, NH HN H

O

H N

H

NH H 2

N-

CI Cl H NH H H

HN

0 o9-- II. Synthesis of the Compounds The compounds of the present invention can be prepared using a variety of procedures, including solid phase and solution phase techniques. A general description of both the solid and solution phase techniques is set forth below. In Section VII, several representative examples are set forth for each of these synthetic techniques.

A. Solid Phase Chemistry Linear Peptide Synthesis: The compounds are synthesized manually (a brief description is set forth in Section VII-C below) or automatically either with a Perkin-Elmer Applied Biosystem Division (PE-ABD) Model 433 automated synthesizer or with a SyntraPrep Reaction Station (from SyntraChem, Charlottesville, VA). All the reagents used for peptide synthesis, can be purchased from PE-ABD. Standard 0.25 mmole FastMoc conductivity monitoring chemistry with single coupling is used with the PE-ABD automated synthesizer. The general Fmoc chemistry protocol for SPPS (solid phase peptide synthesis) includes: 1) cleavage of the Fmoc protection groups with piperidine; 2) activation of the carboxyl group of amino acids; and 3) coupling of the activated amino acids to the amino-terminal of the resin bound peptide chain to form peptide bonds. FastMoc cycles in which amino acids are activated with 2-(lH-benzotriazol- 1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU). 1.0 mmole of dry protected amino acid in a cartridge is dissolved in a solution of HBTU, N,N-diisopropylethylamine (DIEA), WO 02/26774 PCT/US01/30051 and 1-hydroxybenzotriazole (HOBt) in N,N-dimethylformamide (DMF) with additional Nmethylpyrrolidone (NMP) added. The activated Fmoc amino acid is formed almost instantaneously and the solution is transferred directly to the reaction vessel. The step of Fmoc deprotection is monitored and controlled by conductivity measurement. The peptide chain is built on a Rink Amide resin since the C-terminal amide is needed. The acetyl or butyl group is added on the N-terminal side of the peptide after the full length of the peptide chain is made. It is accomplished by reaction of acetic anhydride or butyric anhydride (4.75% V:V acetic anhydride or butyric anhydride, 0.2% HOBt W:V, 2.25% DIEA in NMP) with the ao-amino group on Nterminal side of residue. The final synthesis product is washed extensively with NMP and dichloromethane (DCM). While the SyntraChem reaction station is used for peptide synthesis, O-(7-azabenzotriazol-l-yl)-,l1,3,3-tetramethyluronium hexafluoraphosphate (HATU) is used as an activation reagent to replace HBTU.

Deprotection: The resins containing synthesized peptides are unloaded from the synthesizer and briefly air-dried. Using 4.0-10.0 ml of the cleavage cocktail (91% trifluoroacetic acid (TFA), 2.3% ethanodithiol, 2.3% thioanisol, and 2.3% phenol in water) for 1.5-3.0 hours at room temperature, the peptides are cleaved off the resin and at the same time, the side chain protection groups [O-t-butyl (OtBu) for Asp, Glu, Tyr and Ser, (Pbf) for Arg, t-butoxycarbonyl (Boc) for Trp, Orn, Lys] are removed under the deprotection conditions. The cleavage solution is separated from the resin by filtration. The peptide in the filtrate is then precipitated by adding 40 ml cold ether.

Peptide precipitate is filtered and washed with 4 x 40 ml of cold ether. For the peptides that do not precipitate in the ether solution due to their high degree of hydrophobicity, nitrogen stream is delivered to evaporate the ether. The peptides are then frozen and lyophilized for more than 24hrs. The peptides are recovered into the solution by adding acetic acid.

Purification and Characterization: The peptide powder along with other by-products are re-dissolved in 50% acetic acid solution and injected onto a Vydac 1.0 cm I.D. 25 cm length C-8 column with 5 ~m particle size, and 300 A pore size for purification. A Beckman System Gold HPLC system with dual wavelength u.v. detector is used. Linear gradient of acetonitrile is programmed and introduced to the column to separate the peptide product from other substances.

The elute is collected by a Pharmacia fraction collector, and the individual separation fractions are subjected to both analytical HPLC and electrospray MS for characterization to ensure the identity and purity.

WO 02/26774 PCT/US01/30051 B. Solution Phase Chemistry Solvents and reagents commercially obtained are used without purification. Reaction mixtures are stirred magnetically and are monitored by either analytical high performance liquid chromatography (HPLC) or thin-layer chromatography (TLC). Solutions are routinely concentrated using a Biichi rotary evaporator at 15-25 mm Hg. TLC is performed using silica gel

F

254 precoated plates with a fluorescent indicator. Visualization is accomplished routinely with UV light (254 nm.) Flash column chromatography is carried out with E. Merck silica gel (230-400 mesh) using the eluants indicated; chromatographic separations are monitored by TLC analyses. Analytical HPLC is carried out on either 4.6 x 250 mm MetaChem Kromasil C 4 or Polaris Cg-reverse-phase colunms (3.5 u or 3.0 J particle sizes for C 4 or C 18 respectively) using a 0.1% phosphoric acid in water (A)/acetonitrile gradient B for C 4 or 20% B for Cg 1 to 100 B over 20 min, hold 5 min) with a flow rate of 1.0 ml/min; detection is by UV light at both 214 and 254 nm. Preparative HPLC is conducted on either a 50 x 250 mm Polaris C 18 -reversephase column (10 LI particle size, 100 A pore size) or a 41.4 x 250 mm Rainin Dynamax C 4 reverse-phase column (8 pt particle size, 300 A pore size) using a 0.1% trifluoroacetic acid in water(A)/acetonitrile(B) gradient to 100% B over 55 min, hold 10 min); detection is by UV light at 214 nm.

C. General It is recognized that it is preferable to use a protecting group for any reactive functionality such as a carboxyl, hydroxyl and the like. This is standard practice, well within the normal practice of the skilled artisan.

The indicated steps may be varied to increase yield of desired product. The skilled artisan will recognize the judicious choice of reactants, solvents, and temperatures is an important component in any successful synthesis. Determination of optimal conditions, etc. is routine.

Thus the skilled artisan can make a variety of compounds using the guidance of the above general descriptions, together with the teachings of the examples in Section VII.

It is recognized that the skilled artisan in the art of organic chemistry can readily carry out standard manipulations of organic compounds without further direction; that is, it is well within the scope and practice of the skilled artisan to carry out such manipulations. These include, but are not limited to, reduction of carbonyl compounds to their corresponding alcohols, oxidations of hydroxyls and the like, acylations, aromatic substitutions, both electrophilic and WO 02/26774 PCT/US01/30051 nucleophilic, etherifications, esterification and saponification and the like. Examples of these manipulations are discussed in standard texts such as March, Advanced Organic Chemistry (Wiley), Carey and Sundberg, Advanced Organic Chemistry (Vol. 2) and other art that the skilled artisan is aware of.

The skilled artisan will also readily appreciate that certain reactions are best carried out when potentially reactive functionalities on the molecule are masked or protected, thus avoiding any undesirable side reactions and/or increasing the yield of the reaction. Often the skilled artisan utilizes protecting groups to accomplish such increased yields or to avoid the undesired reactions. These reactions are found in the literature and are also well within the scope of the skilled artisan. Examples of many of these manipulations can be found for example in T. Greene, Protecting Groups in Organic Synthesis. Of course, amino acids with reactive side chains used as starting materials are preferably blocked to prevent undesired side reactions.

IV. Melanocortin Functional Activity and Selectivity Functional activity can be evaluated using various methods known in the art. Examples of such methods are measurement of second messenger responses, in particular cAMP, the use of modified cell systems yielding color reaction upon accumulation of second messenger elements such as cAMP, e.g. as described by Chen et al. 1995 (Anal Biochem. 1995, 226, 349-54), Cytosensor Microphysiometer techniques (see Boyfield et al. 1996), or the study of physiological effects caused by the compounds of the invention may be applied by using the compounds of the invention alone, or in combination with natural or synthetic MSH-peptides.

The compounds of the present invention will interact preferentially selectively) to MC-4 and/or MC-3, relative to the other melanocortin receptors. Selectivity is particularly important when the compounds are administered to humans or other animals, to minimize the number of side effects associated with their administration. MC-3/MC-4 selectivity of a compound is defined herein as the ratio of the EC 50 of the compound for an MC-1 receptor ("ECso-MC-l") over the EC 5 o of the compound for the MC-3 (ECso-MC-3) MC-4 (ECso-MC-4) receptor, the EC 5 0 values being measured as described above. The formulas are as follows: MC-3 selectivity [EC 5 o-MC-1] [ECso-MC-3] MC-4 selectivity [ECso-MC-1] [EC 5 o-MC-4] A compound is defined herein as being "selective for the MC-3 receptor" when the above mentioned ratio "MC-3-selectivity" is at least about 10, preferably at least about 100, and more preferably at least about 500.

WO 02/26774 PCT/US01/30051 A compound is defined herein as being "selective for the MC-4 receptor" when the above mentioned ratio "MC-4-selectivity" is at least about 10, preferably at least about 100, and more preferably at least about 500.

V. Methods of Use and Compositions: Based on their ability to agonize or antagonize the MC-4 and/or MC-3 receptor, the present invention also relates to the use of the ligands described herein in methods for treating obesity and other body weight disorders, including, for example, anorexia and cachexia. The compounds may also be used in methods for treating disorders that result from body weight disorders, including but not limited to insulin resistance, glucose intolerance, Type-2 diabetes mellitus, coronary artery disease, elevated blood pressure, hypertension, dyslipidaemia, cancer endometrial, cervical, ovarian, breast, prostate, gallbladder, colon), menstrual irregularities, hirsutism, infertility, gallbladder disease, restrictive lung disease, sleep apnea, gout, osteoarthritis, and thromboembolic disease. The invention further relates to the treatment of disorders relating to behavior, memory (including learning), cardiovascular function, inflammation, sepsis, cardiogenic and hypovolemic shock, sexual dysfunction, penile erection, muscle atrophy, nerve growth and repair, intrauterine fetal growth, and the like.

The terms treating and treatment are used herein to mean that, at a minimum, administration of a compound of the present invention mitigates a disorder by acting via the MC- 3 or MC-4 receptor. Thus, the terms include: preventing a disease state from occurring in a mammal, particularly when the mammal is predisposed to acquiring the disease, but has not yet been diagnosed with the disease; inhibiting progression of the disease state; and/or alleviating or reversing the disease state.

The invention compounds can therefore be formulated into pharmaceutical compositions for use in treatment or prophylaxis of these conditions. Standard pharmaceutical formulation techniques are used, such as those disclosed in Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa., latest edition and Peptide and Protein Drug Delivery, Marcel Dekker, NY, 1991.

The compositions of the invention comprise: a. a safe and effective amount of a compound of Formula and b. a pharmaceutically-acceptable excipient.

A "safe and effective amount" of a Formula compound is an amount that is effective to interact with the MC-4 and/or MC-3 receptor, in an animal, preferably a mammal, more preferably a human subject, without undue adverse side effects (such as WO 02/26774 PCT/US01/30051 toxicity, irritation, or allergic response), commensurate with a reasonable benefit/risk ratio when used in the manner of this invention. The specific "safe and effective amount" will, obviously, vary with such factors as the particular condition being treated, the physical condition of the patient, the duration of treatment, the nature of concurrent therapy (if any), the specific dosage form to be used, the excipient employed, the solubility of the Formula (1) compound therein, and the dosage regimen desired for the composition.

In addition to the subject compound, the compositions of the subject invention contain one or more pharmaceutically-acceptable excipients. The term "pharmaceutically-acceptable excipient", as used herein, means one or more compatible solid or liquid ingredients which are suitable for administration to an animal, preferably a mammal, more preferably a human. The term "compatible", as used herein, means that the components of the composition are capable of being commingled with the subject compound, and with each other, in a manner such that there is no interaction which would substantially reduce the pharmaceutical efficacy of the composition under ordinary use situations. Pharmaceutically-acceptable excipients must, of course, be of sufficiently high purity and sufficiently low toxicity to render them suitable for administration to the animal, preferably a mammal, more preferably a human being treated.

Some examples of substances which can serve as pharmaceutically-acceptable excipients or components thereof are sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and methyl cellulose; powdered tragacanth; malt; gelatin; talc; solid lubricants, such as stearic acid and magnesium stearate; vegetable oils, such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil and oil of theobroma; polyols such as propylene glycol, glycerin, sorbitol, mannitol, and polyethylene glycol; agar; alginic acid; wetting agents and lubricants, such as sodium lauryl sulfate; coloring agents; flavoring agents; tableting agents, stabilizers; antioxidants; preservatives; pyrogen-free water; isotonic saline; and buffers, such as phosphate, citrate and acetate.

The choice of pharmaceutically-acceptable excipients to be used in conjunction with the subject compound is basically determined by the way the compound is to be administered. If the subject compound is to be injected, the preferred pharmaceutically-acceptable excipient is sterile so water, physiological saline, or mixtures thereof, the pH of which has preferably been adjusted to about 4-10 with a pharmaceutical buffer; a compatible suspending agent may also be desirable.

In particular, pharmaceutically-acceptable excipients for systemic administration include sugars, starches, cellulose and its derivatives, malt, gelatin, talc, calcium sulfate, WO 02/26774 PCT/US01/30051 lactose, vegetable oils, synthetic oils, polyols, alginic acid, phosphate, acetate and citrate buffer solutions, emulsifiers, isotonic saline, and pyrogen-free water. Preferred excipients for parenteral administration include propylene glycol, ethyl oleate, pyrrolidone, ethanol, and sesame oil. Preferably, the pharmaceutically-acceptable excipient, in compositions for parenteral administration, comprises at least about 90% by weight of the total composition.

The compositions of this invention are preferably provided in unit dosage form. As used herein, a "unit dosage form" is a composition of this invention containing an amount of a Formula compound that is suitable for administration to an animal, preferably a mammal, more preferably a human subject, in a single dose, according to good medical practice. These compositions preferably contain from about 1 mg to about 750 mg, more preferably from about 3 mg to about 500 mg, still more preferably from about 5 mg to about 300 mg, of a Formula compound.

The compositions of this invention may be in any of a variety of forms, suitable (for example) for oral, rectal, topical, nasal, ocular, transdermal, pulmonary or parenteral administration. Depending upon the particular route of administration desired, a variety of pharmaceutically-acceptable excipients well-known in the art may be used. These include solid or liquid fillers, diluents, hydrotropes, surface-active agents, and encapsulating substances. Optional pharmaceutically-active materials may be included, which do not substantially interfere with the inhibitory activity of the Formula compound. The amount of excipient employed in conjunction with the Formula compound is sufficient to provide a practical quantity of material for administration per unit dose of the compound.

Techniques and compositions for making dosage forms useful in the methods of this invention are described in the following references, all incorporated by reference herein: Modern Pharmaceutics, Chapters 9 and 10 (Banker Rhodes, editors, 1979); Lieberman et al., Pharmaceutical Dosage Forms: Tablets (1981); and Ansel, Introduction to Pharmaceutical Dosage Forms 2d Edition (1976).

Various oral dosage forms can be used, including such solid forms as tablets, capsules, granules and bulk powders. These oral forms comprise a safe and effective amount, usually at least about and preferably from about 25% to about 50%, of the Formula compound. Tablets can be compressed, tablet triturates, enteric-coated, sugarcoated, film-coated, or multiple-compressed, containing suitable binders, lubricants, diluents, disintegrating agents, coloring agents, flavoring agents, flow-inducing agents, and melting agents. Liquid oral dosage forms include aqueous solutions, emulsions, WO 02/26774 PCT/US01/30051 suspensions, solutions and/or suspensions reconstituted from non-effervescent granules, and effervescent preparations reconstituted from effervescent granules, containing suitable solvents, preservatives, emulsifying agents, suspending agents, diluents, sweeteners, melting agents, coloring agents and flavoring agents.

The pharmaceutically-acceptable excipient suitable for the preparation of unit dosage forms for peroral administration are well-known in the art. Tablets typically comprise conventional pharmaceutically-compatible adjuvants as inert diluents, such as calcium carbonate, sodium carbonate, mannitol, lactose and cellulose; binders such as starch, gelatin, polyvinylpyrrolidone and sucrose; disintegrants such as starch, alginic acid and croscarmelose; lubricants such as magnesium stearate, stearic acid and talc. Glidants such as silicon dioxide can be used to improve flow characteristics of the powder mixture. Coloring agents, such as the FD&C dyes, can be added for appearance. Sweeteners and flavoring agents, such as aspartame, saccharin, menthol, peppermint, and fruit flavors, are useful adjuvants for chewable tablets.

Capsules typically comprise one or more solid diluents disclosed above. The selection of excipient components depends on secondary considerations like taste, cost, and shelf stability, which are not critical for the purposes of the subject invention, and can be readily made by a person skilled in the art.

Peroral compositions also include liquid solutions, emulsions, suspensions, and the like.

The pharmaceutically-acceptable excipients suitable for preparation of such compositions are well known in the art. Typical components of excipients for syrups, elixirs, emulsions and suspensions include ethanol, glycerol, propylene glycol, polyethylene glycol, liquid sucrose, sorbitol and water. For a suspension, typical suspending agents include methyl cellulose, sodium carboxymethyl cellulose, Avicel® RC-591, tragacanth and sodium alginate; typical wetting agents include lecithin and polysorbate 80; and typical preservatives include methyl paraben, propyl paraben and sodium benzoate. Peroral liquid compositions may also contain one or more components such as sweeteners, flavoring agents and colorants disclosed above.

Such compositions may also be coated by conventional methods, typically with pH or time-dependent coatings, such that the subject compound is released in the gastrointestinal tract in the vicinity of the desired topical application, or at various times to extend the desired action.

Such dosage forms typically include, but are not limited to, one or more of cellulose acetate phthalate, polyvinylacetate phthalate, hydroxypropyl methyl cellulose phthalate, ethyl cellulose, Eudragit® coatings, waxes and shellac.

WO 02/26774 PCT/US01/30051 Because the compounds of the present invention are peptidic in nature, a preferred mode of administration is parenteral (more preferably intravenous injection) or nasal administration, in the form of a unit dose form. Preferred unit dose forms include suspensions and solutions, comprising a safe and effective amount of a Formula I compound. When administered parenterally, the unit dose form will typically comprise from about 1 mg to about 3 g, more typically from about 10 mg to about 1 g, of the Formula compound, although the amount of compound administered will depend, for example, on its relative affinity for the MC-4/MC-3 receptor subtypes, its selectivity over other receptors, including the other melanocortin receptors, etc.

Compositions of the subject invention may optionally include other drug actives.

Other compositions useful for attaining systemic delivery of the subject compounds include sublingUal and buccal dosage forms. Such compositions typically comprise one or more of soluble filler substances such as sucrose, sorbitol and mannitol; and binders such as acacia, microcrystalline cellulose, carboxymethyl cellulose and hydroxypropyl methyl cellulose.

Glidants, lubricants, sweeteners, colorants, antioxidants and flavoring agents disclosed above may also be included.

VI. Methods of Administration: As indicated, compositions of this invention can be administered topically or systemically. Systemic application includes any method of introducing a Formula (I) compound into the tissues of the body, intra-articular, intrathecal, epidural, intramuscular, transdermal, intravenous, intraperitoneal, subcutaneous, sublingual, rectal, nasal, pulmonary, and oral administration. The Formula compounds of the present invention are preferably administered systemically, more preferably parenterally and most preferably via intravenous injection.

The specific dosage of compound to be administered, as well as the duration of treatment, and whether the treatment is topical or systemic are interdependent. The dosage and treatment regimen will also depend upon such factors as the specific Formula (I) compound used, the treatment indication, the personal attributes of the subject (such as weight), compliance with the treatment regimen, and the presence and severity of any side effects of the treatment.

Typically, for a human adult weighing approximately 70 kilograms, from about 1 mg to about 6 g, more typically from about 100 mg to about 3 g, of Formula compound are administered per day for systemic administration. It is understood that these dosage ranges WO 02/26774 PCT/US01/30051 are by way of example only, and that daily administration can be adjusted depending on the factors listed above.

As is known and practiced in the art, all formulations for parenteral administration must be sterile. For mammals, especially humans, (assuming an approximate body weight of 70 kilograms) individual doses of from about 0.001 mg to about 100 mg are preferred.

A preferred method of systemic administration is intravenous delivery. Individual doses of from about 0.01 mg to about 100 mg, preferably from about 0.1 mg to about 100 mg are preferred when using this mode of delivery.

In all of the foregoing, of course, the compounds of the invention can be administered alone or as mixtures, and the compositions may further include additional drugs or excipients as appropriate for the indication.

The compound of the invention can be delivered to the preferred site in the body by using a suitable drug delivery system. Drug delivery systems are well known in the art. For example, a drug delivery technique useful for the compounds of the present invention is the conjugation of the compound to an active molecule capable of being transported through a biological barrier (see e.g. Zlokovic, Pharmaceutical Research, Vol. 12, pp. 1395-1406 (1995)). A specific example constitutes the coupling of the compound of the invention to fragments of insulin to achieve transport across the blood brain barrier (Fukuta, et al. Pharmaceutical Res., Vol. 11, pp. 1681-1688 (1994)). For general reviews of technologies for drug delivery suitable for the compounds of the invention see Zlokovic, Pharmaceutical Res., Vol. 12, pp. 1395-1406 (1995) and Pardridge, WM, Pharmacol. Toxicol., Vol. 71, pp. 3-10 (1992).

VII. Representative Synthetic Examples In the following examples, the invention will be described in greater detail by reference to a number of preferred embodiments which are only given for purposes of illustration and should not be considered to limit the invention in any way.

The following abbreviations are used in the Examples: Ac: acetyl [-C(O)CH 3 Aun: aminoundecanoic Ate: (D,L)-2-aminotetraline-2-carboxylic acid Bc: butanoyl [-C(O)(CH 2 2 CH3] Boc: tert-butyloxycarbonyl DCM: dichloromethane DEA: diethylamine DMF: N,N-dimethylformamide DMAP: 4-dimethylaminopyridine DME: 1,2-dimethoxyethane DIEA: diisopropylethylamine DPPA: Diphenylphosphoryl azide EtOAc: ethyl acetate WO 02/26774 PCT/US01/30051 EDCI: 1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide hydrochloride Fmoc: 9-Fluorenylmethoxycarbonyl HOBt: N-hydroxybenzotriazole, monohydrate HOAt: 1-hydroxy-7-azabenzotriazole i-PrOH: 2-propanol MeOH: methanol NMM: N-methylmorpholine OtBu: tert-butoxy [-O-C(CH 3 3 Pbf: 2,2,4,6,7-pentamethyl-dihydrobenzofurane-5-sulfonyl- Pmc: 2,2,5,7,8-pentamethyl-6-chromansulfonylp-TSA: p-toluenesulfonate PyBOP: benzotriazole-lyl-oxy-tris-pyrrolidinophosphonium hexafluorophosphate PyBroP: bromo-tris-pyrrolidino-phosphonium hexafluorophosphate tBu: tert-butyl [-C(CH 3 3 TEA: triethylamine TFA: trifluoroacetic acid THF: tetrahydrofuran A. Automated Solid Phase Chemistry Example 1 Synthesis of Ac-YfRW-NHz Based on the 0.55 mmole/g substitution rate for the Rink Amide resin, 0.45g of the resin is weighted out for 0.25 mmole scale synthesis. The performance of the PE-ABD 433 peptide synthesizer is checked before, the run with various flow tests to ensure the accurate reagent delivery. Fmoc amino acids: Tyr-OtBu, Arg-Pmc, and Trp-Boc are purchased commercially in 1 mmole cartridges. Fmoc-phe (387 mg, 1 mmole) is measured and added in the synthesis cartridges. The freshly made acetic anhydride solution is loaded on the instrument at #4 bottle position. Other synthesis reagents and solvents are purchased commercially and loaded on the instrument according to the instrument's instruction. A chemistry program called NAc- 0.25mmole MonPrePk is used for synthesizing this peptide. The Fmoc deprotection is monitored and controlled by conductivity measurement with set criteria of 5% or less conductivity comparing to previous deprotection cycle.

The resin is air-dried and transferred into a glass vial and a freshly prepared cleavage reagent (10 ml) is added. The deprotection reaction is carried out for 2 hours at room temperature with constant stirring. The supernatant is then separated from the resin by filtration.

The synthesized peptide is then precipitated in ether layer by adding 40 ml cold ether. The peptide precipitates are centrifuged (Heraeus Labofuge 400, Rotor #8179) at 3,500 rpm for four minutes. The ether is discarded and 40 ml of fresh cold ether is added to wash the peptide WO 02/26774 PCT/US01/30051 precipitates. The washing steps are repeated for three times to remove the deprotection byproducts. The final peptide precipitates are freeze-dried overnight. The identity and purity of the linear peptide is determined by both MS and HPLC. Expected peptide molecular weight is detected.

Peptide is re-dissolved in 50% acetic acid and purified by a C8 reverse phase HPLC using a linear gradient of 0-70% solvent B with solvent A in 70 min at a flow rate 3 ml/min. The composition of solvents A and B are as follows: A: 0.1% TFA, 2% acetonitrile in water; B: 0.1% TFA in 95% acetonitrile. The fractions are collected at every 0.25 min. Aliquots of each fraction are analyzed by both MS and analytical RP-HPLC. The fractions that contain a single u.v. 220 nm absorbance peak with expected mass unit for the peptide are combined and lyophilized. The final purity of the peptide is determined by an analytical RP-HPLC of the combined fractions.

The peptides described in Examples 2-54 below are readily synthesized according to the same protocol as Example 1, but with the modifications as noted.

Example 2 Synthesis of Ac-YFRW-NHz Prepared according to Example 1, except Fmoc-L-Phe is used instead Fmoc-D-Phe.

Example 3 Synthesis of Ac-FfRW-NH 2 Prepared according to Example 1, except Fmoc-L-Phe is used instead Fmoc-L-Tyr(OtBu).

Example 4 Synthesis of Ac-PFRW-NH 2 Prepared according to Example 1, except Fmoc-L-Pro is used instead Fmoc-L-Tyr(OtBu).

Example Synthesis of Ac-AfRW-NHz Prepared according to Example 1, except Fmoc-L-Ala is used instead Fmoc-L-Tyr(OtBu).

Example 6 Synthesis of Ac-(2-Nal)fRW-NH2 Prepared according to Example 1, except Fmoc-L-(2-Nal) is used instead Fmoc-L-Tyr(OtBu).

Example 7 Synthesis of Ac-YfR(2-Nal)-NH 2 WO 02/26774 WO 0226774PCT/USOI/30051 Prepared according to Example 1, except Fmoc-L-(2-Nal) is used instead Fmoc-L-Trp(Boc).

Example 8 Synthesis of Ac-YfR(1-Nal)-N112 Prepared according to Example 1, except Fmoc-L-(1-Nal) is used instead Fmoc-L-Trp(Boe).

Example 9 Synthesis of Ae-YfHW-NHz Prepared according to Example 1, except Fmoc-L-HIs(Trt) is used instead Fmoc-L-Arg(Pmc).

Example Synthesis of Ac-Y(D-2-Nal)RW-NH 2 Prepared according to Example 1, except Fmoc-D-(2-Nal) is used instead Frnoc-D-Phe.

Example 11 Synthesis of Ac-Y(L-N-Me-IPhe)RW-NH 2 Prepared according to Example 1, except Fmoc-L-N-Me-Phe is used instead Fmoc-D-Phe.

Example 12 Synthesis of Ac-A(D-N-Me-Phe)RW-N1 2 Prepared according to Example 1, except Fmoc-D-N-Me-Phe is used instead Fmoc-L-Phe, and Fmoc-L-Ala is used instead of Fmoc-L-Tyr(OtBu).

Example 13 Synthesis of Ac-YF(-N-Me-Arg)W-NH Prepared according to Example 1, except Fmoc-L-N-Me-Arg(Mtr) is used instead Fmoc-L- Arg(Pmc), and Fmoc-L-Phe is used instead of Fmoc-D-Phe.

Example 14 Synthesis of Ac-Yf(L-N-Me-Arg)W-N1 2 Prepared according to Example 1, except Pmoc-L-N-Me-Arg(Mtr) is used instead Fmoc-L- Arg(Pmuc).

Example Synthesis of Ac-(L-N-Me-Tyr)FRW-N1J 2 Prepared according to Example 1, except Fmoc-L-N-Me-Tyr(Bzl) is used instead Fmoc-L- Tyr(OtB and Fmoc-L-Phe is used instead of Fmoc-D-Phe.

WO 02/26774 WO 0226774PCT/USO1/30051 Example 16 Synthesis of Ac-(L-N-Me-Tyr)fRW-NH 2 Prepared according to Example 1, except Fmoc-L-N-Me-Tyr(Bzl) is used instead Fmoc-L- Tyr(OtBu).

6 Example 17 Synthesis of Ac-Y(D-4-Chloro-Phe)RW.NH2 Prepared according to Example 1, except Fmoc-D-4-Chloro-Phe is used instead Fmoc-D-Phe.

Example 18 Synthesis of Ac-Y(D-4-Fluoro-Phe)RW-NH 2 Prepared according to Example 1, except Fmoc-D-4-Fluoro-Phe is used instead Fmoc-D-Phe.

Example 19 Synthesis of Ac'.Y(D-3,4-Dichloro-Phe)RW-NH 2 Prepared according to Example 1, except Fmoc-D-3,4-Dichloro-Phe is used instead Fmoc-D-Phe.

Example Synthesis of Ac-Y(D-4-Me-Phe)RW-NHz Prepared according to Example 1, except Fmoc-D-4-Me-Phe is used instead Fmoc-D-Phe.

Example 21 Synthesis of Ac-Y(D-4-Nitro-Phe)RW-NH2 Prepared according to Example 1, except Fmoc-D-4-Nitro-Phe is used instead Fmoc-D-Phe.

Example 22 Synthesis of Ac-Y(D-Phenylglycine)RW-NH 2 Prepared according to Example 1, except Fmoc-D-Phenylglycine is used instead Fmoc-D-Phe.

Examnie 23 Synthesis of Ac-Y(D-4-Homo-Phe)RW-NH 2 Prepared according to Example 1, except Fmoc-D-4-Homo-Phe is used instead Fmoc-D-Phe.

Example 24 Synthesis of Ae-Y(D-Strylryalanine)RW-N1 2 Prepared according to Example 1, except Fmoc-D-Strylryalanine is used instead Fmoc-D-Phe.

Example 41 WO 02/26774 WO 0226774PCT/USOI/30051 Synthesis of Ac-Y(D-4-Thienylalanine)RW-N1 2 Prepared according to Example 1, except Fmnoc-D-4-Thienylalanine is used instead Fmoc-D-Phe.

Example 26 Synthesis of Ac-Y(D-3-Fluoro-Phe)RW-N1 2 Prepared according to Example 1, except Fmoc-D-3-Fluoro-Phe is used instead Fmoc-D-Phe.

Example 27 Synthesis of Ac.(L-4-Fluoro-Phe)(D-4-Fluoro-Phe)RW-NH 2 Prepared according to Example 1, except Fmoc-L-4-Fluoro-Phe is used instead Fmoc-L- Tyr(OtB and Fmoc-D-4-Fluoro-Phe is used instead of Fmoc-D-Phe.

Example 28 Synthesis of Ac-Y(D-2-Fluoro-Phe)RW-NIz Prepared according to Example 1, except Fmioc-D-2-Fluoro-Phe is used instead Fmoc-D-Phe.

Example 29 Synthesis of Ac-(L-4-Chloro-Phe)(D-4-Fluoro-Phe)RW-NH 2 Prepared according to Example 1, except Fmoc-L-4-Chloro-Phe is used instead Fmoc-L- Tyr(OtBu), and Fmoc-D-4-Fluoro-Phe is used instead of Fmoc-D-Phe.

Example Synthesis of Ac-(L-4-Chloro-Plie)(D-4-Fluoro-Phe)RW(N-Me-Gly)-N1 2 Prepared according to Example 1, except Fmoc-L-4-Chloro-Phe is used instead Fmoc-L- Tyr(OtBu), Fmoc-D-4-Fluoro-Phe is used instead of Fmoc-D-Phe, and an additional Fmoc-N-Me- Gly is used.

Example 31 Synthesis of Ac-(L-4-Chloro-Phie)fRW(N-Me-Gly)-N1 2 Prepared according to Example 1, except Frnoc-L-4-Chiloro-Phe is used instead Fmoc-L- Tyr(OtBu), and an additional Fmoc-N-Me-Gly is used.

Example 32 Synthesis of Ac- (L-4-Chloro-Phe)4D-4-Flijoro-Phe)R-NH 2 WO 02/26774 WO 0226774PCT/USOI/30051 Prepared according to Example 1, except Fmoc-L-4-Cliloro-Phe is used instead Fmoc-L- Tyr(OtBu), Fmoc-D-4-Fluoro-Phe is used instead of Fmoc-D-Phe, and Fmoc-L-Trp(Boc) is not used.

Example 33 Synthesis of Ac-(L-4-Chloro-Phe)(D-4-Fluoro-Phe)RG-Nz Prepared according to Example 1, except Fmoc-L-4-Chloro-Phe is used instead Fmoc-L- Tyr(OtBu), Fmoc-D-4-Fluoro-Phe is used instead of Fmoc-D-Phe, and Fmoc-L-Gly is used instead of Fmoc-L-Trp(Boc).

Example 34 Synthesis of Ac-(L-3,4-Difluoro-Phe)fRW-NIHz Prepared according to Example 1, except Fmoc-L-3,4-Difluoro-Phe is used instead Fmoc-L- Tyr(OtBui).

Example Synthesis of Ac-Y(D-2-Me-Phe)RW-NHz Prepared according to Example 1, except Fmoc-D-2-Me-Phe is used instead Fmoc-D-Phe.

Example 36 Synthesis of Ac-(L-4-Bromo-Phe)fRW-NH 2 Prepared according to Example 1, except Fmoc-L-4-Bromo-Phe is used instead Fmoc-L- Tyr(OtBu).

Example 37 Synthesis of Ac-(L-4-Iodo-Phe)fRW-N1 2 Prepared according to Example 1, except Fnmoc-L-4-Iodo-Phe is used instead Fmoc-L-Tyr(OtBu).

Example 38 Synthesis of Ac-(L-Pentaflnoro-Phe)fRW-N1 2 Prepared according to Example 1, except Fmoc-L-Pentafluoro-Phe is used instead Fmnoc-L- Tyr(OtBu).

Example 39 Synthesis of Ac-(L-4-Nitro-Phe)fRW-N1 2 WO 02/26774 WO 0226774PCT/USOI/30051 Prepared according to Example 1, except Fnaoc-L-4-Nitro-Phe is used instead Fmoc-L- Tyr(OtBu).

Example Synthesis of Ac-(L-Asninomethyl-Phe)fRW-NH 2 Prepared according to Example 1, except Fmoc-L-Aminomethyl-Phe(Boc) is used instead Fmoc- L-Tyr(OtBu).

Example 41 Synthesis of Ac-(L-Tetraisoquinoline-3-carboxylic acid)fRW-NH 2 Prepared according to Example 1, except Fmoc-L-Tetraisoquinoline-3-carboxylic acid is used instead Fmoc-L-Tyr(OtBu).

Example 42 Synthesis of Ac-(L-Homo-Phe)fRW-NI~z Prepared according to Example 1, except Fmoc-L-Homo-Phe is used instead Fmoc-L-Tyr(OtBu).

Example 43 is Synthesis of Ac-(L-Biphenyl-Alanine)RW-NH 2 Prepared according to Example 1, except Fmoc-L-Biphennyl-Alanine is used instead Fmoc-L- Tyr(OtBu).

Example 44 Synthesis of Ac-(L-4"S0 3 -Phe)fRW-N{ 2 Prepared according to Example 1, except Fmoc-L-4-S0 3 -Phe is used instead Fmoc-L-Tyr(OtBn).

Example Synthesis of Ac-(L-2,6-Dimnethyl-Phe)fRW-NH 2 Prepared according to Example 1, except Fmoc-L-2,6-Dimethyl-Phe is used instead Fmoc-L- Tyr(OtBu).

Example 46 Synthesis of Ac-(L-4-Methyl-Phe)fRW-NH 2 Prepared according to Example 1, except Fmoc-L-4-Methyl-Phe is used instead Fmoc-L- Tyr(OtBu).

WO 02/26774 WO 0226774PCT/USOI/30051 Example 47 Synthesis of Ac- (L-4-N-H-Phe)fRW-NH 2 Prepared according to Example 1, except Fmoc-L-4-NII-Phe(B3oc) is used instead Finoc-L- Tyr(Otflu).

Example 48 Synthesis of Ac-YfKW-NR 2 Prepared according to Example 1, except Fmoc-L-Lys(Boc) is used instead Fmoc-L-Arg(Pmc-).

Example 49 Synthesis of Ac-Yf(Orn)W-NH2z Prepared according to Example 1, except Fmoc-L-Om(Boc) is used instead Fmoc-L-Arg(Pmc).

Example Synthesis of Bc-HFRW(N-Me-Gly)-NH 2 Prepared according to Example 1, except Fmoc-L-Hils(Trt) is used instead Fmoc-L-Tyr(OtBu), Fmoc-L-Phe is used instead of Fmoc-D-Phe, butyric anhydride is used instead of acetic anhydride, and an additional Fmoc-L-N-Me-Gly is used.

Example 51 Synthesis of Bc-HfRWV(N-Me-Gly)-NH2 Prepared according to Example 1, except Fmoc-L-Hlis(Trt) is used instead Fmoc-L-Tyr(OtBu), butyric anhydride is used instead of acetic anhydride, and an additional Fmoc-L-N-Me-Gly is used.

Example 52 Synthesis of Bc-YfRW(N-Me-Gly)-NH2 Prepared according to Example 1, except butyric anhydride is used instead of acetic anhydride, and an additional Fmoc-L-N-Me-Gly is used.

Example 53 Synthesis of Bc-YFRW(N-Me-Gly)-N1 2 Prepared according to Example 1, except Fmoc-L-Phc is used instead of Fmloc-D-Phe, butyric anhydride is used instead of acetic anhydride, and an additional Fmoc-L-N-Me-Gly is used.

Example 54 WO 02/26774 WO 0226774PCT/USOI/30051 Synthesis of Bc-FfRW(N-Me-GIy)-NHz, Prcparcd according to Example 1, except Fmoc-L-Phe is used instead Fmoc-L-Tyr(OtBu), butyric anhydride is used instead of acetic anhydride, and an additional Fmoc-L-N-Me-Gly is used.

B. Solution Phase Chemistry Example Synthesis of 5-Guanidino-2-[3-phenyl-2-(3-phenyl-propylamino)-propionylaminopentanoic acid (naphthalen-1-ylmethyl)-amide ii:: i1 2-(S)-(2-(R)-tert-B3utoxycarbonylamino-3-pheyl-propionylamino)-5-nitroguanidinopentanoic acid metl ester To a solution of 2-(R)-(2-tert-butoxycarbonylamino-3-phenyl-propionic acid (2.0g, 7.54 mmoles) and 2-(S)-amino-5-nitroguanidino-pentanoic acid methyl ester (2.0g, 1.leq) in lO0rni of anhydrous DMIF is added 1-[3-(dimethylamino)propyl)-3-ethylcarbodiimide hydrochloride 1-hydroxybenzotriazole (2.l6g, l.4eq) and triethylamine (3.Oml, 3eq). The resulting solution is stirred at room temperature for 20 -hours, and then the solvents are removed under reduced pressure. The resulting residue is partitioned between 10% sodium carbonate (lO0ml) and methylene chloride (lO0mi). The organics are dried over anhydrous magnesium sulfate, filtered and the solvent removed under reduced pressure. The resulting crude material is purified by flash chromatography on silica gel (90:9:1 chloroform:methanol: ammonium hydroxide) to afford the title compound.

2-(S)-(2-(R)-tert-butoxvcarbonylamino-3-:phenvl-prgoniylarino)-5-nitroguanidinouentanoic acid To a solution of 2-(S)-(2-(R)-tert-butoxycarbonylarnino-3-phenyl-propionylamino)-5nitroguanidino-pentanoic acid methyl. ester (3.6g, 7.5mmoles) in tetrahydrofuran (lO0mir) is added lithium hydroxide monohydrate (387mg, 1.2eq) and water (l1nil). After stirring at room temperature for one hour, the reaction is neutralized with trifluoroacetic acid (0.7rnl) and the solvents are removed under reduced pressure. The resulting residue is partitioned between water (200m1) and ethyl acetate (200nm1). The aqueous layer is extracted with ethyl acetate (3 x 250m1), 46 WO 02/26774 WO 0226774PCT/USOI/30051 the organics are pooled and dried over anhydrous magnesium sulfate, filtered and the solvent removed under reduced pressure. The crude material is used without further purification.

(1b(S)-f 4-Nitroguanidinio-l-[naphthalen-l-yl-methvl)-carbamoyll -butylcarbamoyl l-2-(R)-phenvlety-carbamic acid tert-butyl ester To a solution of 2-(S)-(2-(R)-tert-butoxycarbonylan-ino-3-phenyl-propionylaniino)-5-guanidinopentanoic acid 1.0g, 2.15 mmoles) and C-naphthalen-1 -yl-methylantine (0.377n1, 1.2eq) in of anhydrous DMF is added l-[3-(dimethylamino)propyl)-3-ethylcarbodiimide hydrochloride (532mg, 1.5eq), 1-hydroxybenzotriazole (376, L.4eq) and triethylainiine (0.9m1A, 3eq). The resulting solution is stirred at room temperature for 20 hours, and then the solvents are removed under reduced pressure. The resulting residue is partitioned between 10% sodium carbonate (75m1A) and chloroform (75m1). The organics are dried over anhydrous magnesium sulfate, filtered and the solvent removed under reduced pressure. The resulting crude material is purified by flash chromatography on silica gel (90:9:1 chloroformn:methanol:ammoniumhlydroxide) to afford the title compound.

2-(S)-2-(R)-Amino-3-:phenyl-proionylanmino)-5-nitrogruanidino-pentanoic acid (naphthalen-1ylmethyl)-ainide trifluoroacetic acid salt To a solution of 4-nitroguanidino-1-[naphithalen- l-ylmethyl-carbamoyl]butylcarbamoyl}-2-(R)-phenyl-ethyl)-carbainic acid tert-butyl ester 1 .1g, l.82inmoles) in methylene chloride (lO0mI) is added trifluoroacetic acid (50mI). The resulting solution is stirred at room temperature for three hours and the then the solvents are removed under reduced pressure. The crude material is purified by reverse phase preparative HPLC to afford the title compound.

5-Nitroguaiiidino-2-r3-phenyl-2-(3-nhenyl-propylamino)prouionvlaino-pentanoic acid (ngphthalen-1-ylmethyl)-arnide To a suspension of 2-(S)-(2-(R)-aminio-3-phenyl-propioniylamino)-5-guanidino-pentanoic acid (naphthalen-1 -ylmethyl)-amide (750mg, 1.21 mmoles), 3-phenyl-propionaldehyde 159m1, l.Oeq), and activated molecular sieves (4 angstrom, crushed) is added triethylamine (0.247m1l, 1 .5eq). The resulting suspernsion is stirred at room temperature for 24 hours, and then the pH is adjusted to 5 with acetic acid. A LOM solution of sodium cynanoborohydride in tetrahydrofuran (l.44nil, 1.2eq) is then added at a rate of O.2nil/min with a syringe pump. The resulting suspension is stirred at room temperature for 24 hours, filtered through celite, and the solvents WO 02/26774 PCT/US01/30051 removed under reduced pressure. The crude material is purified by reverse-phase HPLC to afford the title compound.

5-Guanidino-2-[3-phenyl-2-(3-phenvl-propylamino)-propionylaminol-pentanoic acid (naphthalen-1 -lmethyl)-amide To a solution of 5-nitroguanidino-2-[3-phenyl-2-(3-phenyl-propylamino)-propionylamino]pentanoic acid (naphthalen-l-ylmethyl)-amide (140mg, 0.165mmoles) in methanol (30ml) is added acetic acid (3ml) and 5% palladium on barium sulfate (100mg). The resulting suspension is hydrogenated at atmospheric pressure for twenty-four hours, and then filtered through celite.

The solvents are removed under reduced pressure and the crude product purified by reverse-phase

HPLC.

Example 56 Synthesis of 5-Guanidino-2-(S)-[3-phenyl-2-(R)-propylamino)-propionylamino]-pentanoic acid (2-naphthalen-2-yl-ethyl)-amide -i I H N Toluene-4-sulfonic acid 2-naphthalen-2yl-ethyl ester To a solution of 2-naphthalen-2yl-ethanol (3.0g, 17.4mmoles) in tetrahydrofuran (50ml), is added para-toluenesulfonic anhydride (6.8g, 1.2eq), and triethylamine (7.1ml, 3eq). The resulting solution is stirred at room temperature for one hour and then the solvents are removed under reduced pressure. The crude material is purified by flash chromatography on silica gel ethyl acetate/hexane) to afford the title compound.

2-(2-Azido-ethyl)-naphthalene To a solution of toluene-4-sulfonic acid 2-naphthalen-2yl-ethyl ester (5.0g, 15.3mmoles) in DMF (100ml) is added sodium azide (1.3g, 1.3 eq). The resulting suspension is heated to 80 0 C for twenty-four hours and then cooled to room temperature. The solvent is removed under reduced pressure and the residue partitioned between ethyl acetate (200ml) and water (200ml). The organics are dried over anhydrous magesium sulfate, filtered and the solvent removed under WO 02/26774 WO 0226774PCT/USOI/30051 reduced pressure to afford the title compound. The crude material is used without further purification.

2-Naphthalen-2-yl-ethylamine To a solution of 2-(2-azido-ethyl)-naphthalene (3.0g, 15.2nimoles), in tetrahydrofuran (lO0mi) is added triphenyiphosphine (6.0g, 1.5eq) and water (5rnl). The resulting solution is heated to reflux for three hours and then cooled to room temperature. The solvents are removed under reduced pressure and the crude material purified by flash chromatography on silica gel (90:9:1 chloroform:m-ethanol:amnnonium hydroxide). The purified material is then converted to the trifluoroacetic acid salt by the addition of excess trifluoroacetic acid, and subsequent removal of the excess acid by evaporation under reduced pressure to afford the title compound.

I 1-(S)-r4-Nitroguanidino-1-(2-naphthalen-2yl-ehylcarbamovl)butylcarbamol-2-(R)-phenvlethyl I -carbamic acid tert-butyl ester To a solution of 2-(S)-(2-(R)-tert-butoxycarbonylamino-3-phenyl-propionylamino)-5nitroguanidinio-pentanoic acid (1.0g, 2. l4irnuoles) and 2-(2-azido-etliyl)-naphtlialenie (733mg, 1 .2eq) in DIVI (50m1) is added l-[3-(dimethylam-ino)propyl)-3-ethylcarbodiimide hydrochloride (6 14mg, 1.5eq), 1-hydroxybenzotriazole (434mg, l.5eq), and triethylamine (0.877ni1, 3eq). The resulting suspension is stirred at room temperature for twenty-four hours, and the solvents are then removed tinder reduced pressure. The crude material is partitioned between 10%/ sodium carbonate (75m1) and methylene chloride (75m1). The organics are dried over anhydrous magnesium sulfate, filtered and solvents removed under reduced pressure. The crude material is purified by flash chromatography on silica gel (90:9:1 chloroform:methanol: ammonium hydroxide) to afford the title compound.

2-(S)-(2-(R)-Armino-3-phen-vl-vropionvlamino)-5-nitroguanidino-nentanoic acid (2-naphthalen-2yl-ptlivl)-amide To a solution of l-(S)-[4-nitroguanidino-1-(2-naphthalen-2-yl-ethylcarbamoyl)-butycarbamoyl- 2-(R)-phenyl-ethyl 1-carbamic acid tert-butyl ester (1 .5g, 2.O4mmoles) in methylene chloride (lO0mI) is added trifluoroacetic acid (50m1). The resulting solution is stirred at room temperature for twenty-four hours and then the solvents are removed tinder reduced pressure. The crude material is purified by flash chromatography on silica gel (90:9:1 chloroform:methanol:am-monium hydroxide) to afford the title compound.

5-Nitroguanidino-2-(S)-r3-phenyl-2-(R):nropvlamino):propionylamiliolzpentanoic acid (2na-phthalen-2-YI-ethfl)-aniide WO 02/26774 WO 0226774PCT/US01/30051 To a solution of 2-(S)-(2-(R)-aniino-3-phenyl-propionylam-ino)-5-nitroguanidino-pentanoic acid (2-naphthalen-2-yl-ethyl)-amide (600mg, 1. l5mmoles) in tetrahydrofuran (5Oinl) is added 3phenyl-propionaldehyde (0.l2lnil, O.8eq) and molecular sieves (4angstrom, crushed). The resulting suspension is stirred at room temperature for twenty-four hours and then the pH is adjusted to 5 with acetic acid. To this solution is added sodium cyanoborohydride (1.3 Smi, 1.2eq, 1 OM solution in tetrahydrofuran) at rate of O.2mllhour using a syringe pump. The resulting suspension is stirred at room temperature for twenty-four hours, and then the solvents are removed under reduced pressure. The crude material is filtered and then purified by reversephase HPLC to afford the title compound.

5-Guanidino-2-(S)-3-ph-eny1-2-(R)-propylainino)propiovylamino1-pentanioic acid (2-nahthalen- 2-yl-etl)-amide To a solution of 5-niitroguanidino-2-(S)-r3-phenyl-2-(R)-propylamino)-propionylarmino ipentanoic acid (2-naphthalen-2-yl-ethyl)-amide (294mg, O.46mmoles) in 50m1 of methanol is added acetic acid (5mil) and 5% palladium on barium sulfate (294mg). The resulting suspension is hydrogenated at atmospheric pressure for twenty-four hours, filtered through celite, and the solvents removed under reduced pressure. The crude material is purified by reverse-phase HPLC to afford the title compound.

Example 57 Synthesis of 5-Guanidino-2-(S)-[3-phenyl-2-(R)-(3-phenyl-propylam-ino)-propionylaminolpentanoic acid (2-naphthalen-1-yl-ethyl)-amide N 0H Toluene-4-sulfonic acid 2-naphthalen-l:-Yl-ethyl ester To a solution of 2-naphthalen-1-yl-ethanol (3.0g, l7.4nimoles) in tetrahydrofuran (50mI) is added para-toluene sulfonic anhydride (6.8g, 1.2eq) and triethylaruine (7.lirn, 3eq). The resulting solution is stirred at room temperature for one hour and then the solvents are removed under WO 02/26774 WO 0226774PCT/USOI/30051 reduced pressure. The crude product is purified by flash chromatography on silica gel (20% ethyl acetatelhexanes) to afford the title compound.

1-(2-Azido-ethyl)-naphthalene To a solution of toluene-4-sulfonic acid 2-naphthalen-l-yl-ethyl ester (5.2g, 15.9rnmoles) in DMF (l0Oml) is added sodium azide (1.3g, 1.3eq). The resulting solution is heated to 80'C to twentyfour hours and then cooled to room temperature and the solvents removed under reduced pressure. The residue is partitioned between ethyl acetate and water, the organics are dried over anhydrous magnesium sulfate, filtered, and the solvents removed under reduced pressure to afford the title compound. The crude product is used without further purification.

2-Naphthalen-1-yl-ethylami-ne To a solution of 1-(2-azido-ethyl)-naphthalene (3.0g, 15.23mmoles) in tetrahydrofuran (lO0mi) is added triphenyiphosphine (6.0g, l.5eq) and water (5m1). The resulting solution is refluxed for three hours and then cooled to roome temperature. The solvents are removed under reduced pressure and the crude product is purified by flash chromatography on silica gel (90:9:1 chlorofonn:methanol:ammoniurn hydroxide) to afford the title compound. The addition of excess trifluoroacetic acid, followed by evaporation provided the trifluoroacetic acid salt.

41-(S)-4-nitroguanidinco-1-2-nphthalen-l-yl-ehlcarbamoyl)-butvlcarbamoyll-2-(R):ph-enlethyl] -carbamic acid tert-butyl ester To a solution of 2-(S)-(2-(R)-tert-butoxycarbonylamino-3-phenyl-propionylamino)-5nitroguanidino-pentanoic acid methyl ester (1 .0g, 2. inunoles) and 2-naphthalen- 1-yl-ethylamine (733mg, 1.2eq) in DMP (50m1) is added 1-hydroxybenzotriazole (434 mg, 1.5eq), 1-[3- (dimethylaminopropyll-3-ethylcarbodiirnide hydrochloride (6 14mg, 1 .5eq), and triethylamine (0.877m1, 3eq). The resulting suspension is stirred at room temperature for twenty-four hours, and then the solvents are removed under reduced pressure. The residue is partitioned between methylene chloride and 10% sodium carbonate, the organics dried over anhydrous magnesium sulfate, filtered and the solvent removed under reduced pressure to give the crude product. The product is purified by flash chromatography on silica gel (90:9:1 chloroform: methanol: ammonium hydroxide) to afford the title compound.

2-(S)-(2-(R)-Amino-3-uhe~nyl-uronionlanmino)-5-nitroauanidino-nentanoic acid (2-naphithalen-lyl-etl)-amide To a solution of I 1-(S)-[4-nitroguanidino-1-(2-naphthalen-1-yl-ethylcarbamoyl)-butylcarbamoyll- 2-(R)-phenyl-ethyl]-carbamic acid tert-butyl ester (1-3g, 1.77nimoles) in methylene, chloride WO 02/26774 PCT/US01/30051 (100ml) is added trifluoroacetic acid (50ml). After stirring at room temperature for twenty-four hours the solvents are removed under reduced pressure ant the residue partitioned between sodium carbonate (100ml) and ethyl acetate (100ml). The organics are dried over anhydrous magnesium sulfate, filtered, and the solvents removed under reduced pressure. The crude product is purified by flash chromatography on silica gel (90:9:1 chloroform:methanol:ammonium hydroxide) to give the title compound.

5-Nitroguanidino-2-(S)-r3-phenyl-2-(R)-(3-phenyl-propylamino)-propionylaminol-pentanoic acid (2-naphthalen-1-vl-ethyl)-amide To solution of 2-(S)-(2-(R)-aiino-3-phenyl-propionylamino)-5-nitroguanidino-pentanoic acid (2naphthalen-l-yl-ethyl)-amide (690mg, 1.33mmoles) in tetrahydrofuran (501ml) is added 3-phenylpropionaldehyde (0.14ml, 0.8eq) and molecular selves (500mg, 4A powdered). The resulting suspension is stirred at room temperature for twenty-four hours and then the pH is adjusted to with acetic acid. Sodium cyanoborohydride (1.6ml 1.2eq 1.OM solution in tetrahydrofuran) is then slowly added (0.2ml/hour) with a syringe pump. After twenty-four hours the solvents are is removed under reduced pressure and the crude product is purified by reverse phase HPLC to afford the title compound.

5-Guanidino-2-(S)-[3-phenyl-2-(R)-(3-phenyl-propylamino)-propionyamino -pentanoic acid (2naphthalen-1-vl-cthyl)-amide To solution of 5-nitroguanidino-2-(S)-[3-phenyl-2-(R)-(3-phenyl-propylamino)-propionylamino]pentanoic acid (2-naphthalen-l-yl-ethyl)-amide (280mg, 0.44moles) in 50 ml of methanol is added acetic acid (5ml) and 5% palladium on barium sulfate (280mg). The resulting suspension is hydrogenated at atmospheric pressure for twenty-four hours, filtered through celite, and the solvents are removed under reduced pressure. The crude material is purified by reverse-phase HPLC to afford the title compound.

Example 58 Synthesis of 5-Guanidino-2-(S)-(2-(S)-{[3-(4-hydroxy-phenyl)-propionyl]-methyl-amino}-3phenyl-propionylamino)-penta acid [2-(1H-indol-3yl)-ethyl]-amide WO 02/26774 WO 0226774PCT/USOI/30051 2-(S)-[2-(S)-(tert-Butoxycarbonyl-methvl-anmino)-3-phenyl-propionylamlinol-5-nitrog-uanidinopentanoic acid methyl este To a solution of 2-(S)-(tert-butoxycarbonyl-methyl-amino)-3-phenyl-propionic acid 7. l7nirnoles) and 2-(S)-amuino-5-nitroguanidino-pentanoic acid methyl ester (1.83g, 1.leq) is added 1-hydroxybenzotriazole (1 .45g, 1 .5eq), 1 -[3-(dimethylamrinopropyl] -3-ethylcarbodiim-ide hydrochloride (2.05g, 1.5eq), and triethylamine (3.OmlA, 3eq). The resulting solution is stirred at room temperature for twenty-four hours, and then the solvents are removed under reduced pressure. The residue is partitioned between 10% sodium carbonate (l5Ornl) and methylene chloride (150n11). The organics are dried over anhydrous magnesium sulfate, filtered and the solvents removed under reduced pressure. The crude material is purified by flash chromatography on silica gel (90:9:1 chloroform: methanol: ammonium hydroxide) to give the title compound.

5-Nitroguanidino-2-(S)-(2-(S)-methvlanino-3-phenvl-prLopionlanmino):-nentanoic acid methyl ester To a solution of 2-(S)-[2-(S)-(tert-butoxycarbonyl-methyl-amino)-3-pbenyl-propionylam-ino]-5nitroguanidino-pentanoic acid methyl ester (3.3g, 6.68mmoles) in methylene chloride (200m1) is added trifluoroacetic acid (lO0mi). The resulting solution is stirred at room temperature for twenty-four hours, and then the solvents are removed under reduced pressure. The residue is partitioned between methylene chloride (15Ornl) and 10% sodium carbonate (150m1l). The organics are dried over anhydrous magnewium sulfate, filtered, and the solvents removed under reduced pressure. The crude material is purified by flash chromatography on silica gel (90:9:1 chloroformn:methanol:ammonium hydroxide) to afford the title compound.

5-Nitroguanidino-2-(S)-(2-(S)-f [3-(4-hydroxvohenyl)prop~ionll-methyl-amino I -3-uhenylpropionylamino)-pentanoic acid methyl estr To a solution of 5-Guanidino-2-(S)-(2-(S)-methylamino-3-phenyl-propionylamino0-pentanoic acid methyl ester (1.0g, 2.54mmoles) and 3-(4-hydroxyphenyl)-propanoic acid (506mg, l.2eq) is WO 02/26774 WO 0226774PCTIUSOI/30051 added 1-hydroxybenzotriazole (513mg, 1.5eq), l-[3-(dimethylainopropyl]-3-eth-ylcarbodiimide hydrochloride (727mg, 1.5eq) and triethylaniine (I.Om1, 3eq). The resulting solution is stirred at room temperature for twenty-four hours, and then the solvents are removed under reduced pressure. The residue is partitioned between 10% sodium carbonate (lO0nil) and methylene chloride (lO0mi). The organics are dried over arnhydrous magnesium sulfate, filtered, and the solvents removed under reduced pressure. The crude material is purified by flash chromatography on silica gel (90:9:1 chlnrofon-n:methanol:ar-nmoniuni hydroxide) to afford the title compound.

5-Nitroguanidino-2-(S)-2-SH- E34(4 h droxhenyfpoinl 1-methyl-amino l-3-phenIl propionlamino)-:pentanoic acid To a solution of 5-nitroguanidino-2-(S)-(2-(S)-{ [3-(4-hydroxyphenyl)propionyl]-metliyl-ainino 1- 3-phenyl-propionylamino)-pentanoic acid methyl ester (5 34mg, 0.9 8mrnoles) in tetrahydrofuran is added lithium hydroxide monohydrate (49mg, 1.leq), and water The resulting solution is stirred at room temperature for twenty-four hours and then acidified with trifluoroacetic acid (leq). The solvents are removed under reduced pressure and the crude material partitioned between ethyl acetate (75m1) and water (75m1). The organic s are dried over anhydrous magnewium sulfate, filtered, and the solvents removed under reduced pressure to afford the title compound.

5-Nitrog4uanidino-2-(S)-(2-(S)-f r3-(4-hydroxy-phenyl)-propionyl]-methyl-amnino 1-3-phenylIprgpionylamino)-p2enta acid r2-(lH-indol-3v1)-cthvlI-amide To a solution of 5-niitroguanidino-2-(S)-(2-(S)-{ [3-(4-hydroxyphenyl)propionylj-methyl-aininol 3-phenyl-propionylamino)-pentanoic acid (460mg, 0. 87mmoles) and [2-(1H-indol-3y1)-ethyl]amine, (168mg, 1.2eq) is added 1-h-ydroxybenzotriazole (176mg, l.5eq), 14[3- (dimethylaminopropyl]-3-ethylcarbodiimide hydrochloride (25 0mg, 1.5eq) and triethylamine (0.36m1A, 3eq). The resulting solution is stirred at room temperature for twenty-four hours, and then the solvents are removed under reduced pressure. The residue is partitioned between ethyl acetate (75m1) and water (75mi), the organics dried over magnesium sulfate, filteed and the solvents removed under reduced pressure. The crude product is purified be reverse-phase HPLC to afford the title compound.

3o 5-Guanidino-2-(S)-(2-(S)-{ [3-(4-hydroxy-:phenyl)-:prop2ionvll-mcthyl-amino I-3-phenylpropionylanino)-:penta acid r2-( 1H-indol-3v1)-ethylj-amnide WO 02/26774 WO 0226774PCT/USOI/30051 To solution of 5-nitroguanidino-2-(S)-(2-(S)-{ [3-(4-hydroxy-phenyl)-propionylj-methyl-ainino}- 3-phenyl-propionylamino)-penta acid [2-(1H-indol-3y1)-ethyll-amide (250mg, 0.37inoles) in ml of methanol is added acetic acid (5nil) and 5% palladium on barium sulfate (25 0mg). The resulting suspension is hydrogenated at atmospheric pressure for twenty-four hours, filtered through celite, and the solvents removed under reduced pressure. The crude material is purified by reverse-phase HPLC to afford the title compound.

Example 59 Synthesis of 5-Guianidino-2-(S)-[3-phenyl-2-(R)-(3-phenyl-propylamino)-propionyaniino]pentanoic acid benzyl amide

NN

HN

N

1-Benzylcarbamoyl-4-nitroguanidino-butvcarbamoyl)-2-(R)-:pheny-ethyll-carbarmic acid tert-butyl este To a solution of 2-(S)-(2-(R)-tert-butoxycarbonylamino-3-phenyl-propionylamino)-5nitroguanidino-pentanoic acid (500mg, 1.29mmoles) benzylamine (0.155m], 1.leq) in DM (50ro) is added l-hydroxybenzotriazole (261mg, 1.5eq), 1-[3-(dimethylamiinopropyl]-3ethylcarbodiiniide hydrochloride (371mg, 1.Seq) and triethlylaiine (0.5 3rl, 3eq). The resulting solution is stirred at room temperature for twenty-four hours, and then the solvents are removed under reduced pressure. The crude material is purified by flash chromatography on silica gel (90:9: 1 chloroform:methanol: ammonium hydroxide) to afford 526mng the title compound.

2-(S)-(2-(R)-Arnino-3-pheniyl-:propionylarnino)-5-niitroguanidino-opentanoic acid benzylam-ide To a solution of [I -(S)-(1-benzylcarbamoyl-4-nitroguanidino-butylcarbamoyl)-2-(R)-phenylethyl] -carban-ic acid tert-butyl ester (512mg, l.l2mnoles) in methylene chloride (50nml) is added trifluororacetic acid (25m1). The resulting solution is stirred at room temperature for twenty-four hours, and the solvents are then removed under reduced pressure. The crude product is purified by reverse-phase HPLC to afford the title compound.

5-Nitroguanidino-2-(S)-[3-:phenyl-2-(Rh-(3:-he nvl-vropvylamino)vroionvlaino1-pentanoic acid benzyl amide WO 02/26774 WO 0226774PCT/USOI/30051 To suspension of 2-(S)-(2-(R)-amino-3-phenyl-propionylamino)-5-nitoguanidino-pentanoic acid benazylamnide (370mg, 0.65mmoles), 3-phenyl-propionaldehyde (0.077n21, 0.9eq) and molecular seives (370mg, 4A powdered) is added triethylamine 177in1, 2eq). The resulting suspension is stirred at room temperature for twenty-four hours, and then the pH is adjusted to 5 with acetic acid. Sodium cyanoborohydride (O.70m1, lOM solution in tetrahydorofuran, leq) is then slowly added (0.2inllhour) with a syringe pump. After twenty-four hours the suspension is filtered through celite and the solvents removed under reduced pressure. The crude product is purified by reverse-phase HPLC to afford the title compound.

5-Guanidino-2-(S)-E3-phenyl-2-(R)-(3zphenyl-propylarmino)-propiony)lainino -pentanoic acid benzyl anfide To a solution of 5-nitroguaniidino-2-(S)-[3-phenyl-2-(R)-(3-phenyl-propylamino)propionylamino]-pentanoic acid benzyl amide (80mg, 0. l4rnmoles) in methanol (50Rmi) is added acetic acid (5mi) and 5% palladium on abrium sulfate (80mig). The resulting suspension is hydrogenated at atmospheric pressure for twenty-four hours, filtered through celite and the solvents removed under reduced pressure. The crude product is purified by reverse-phase BPLC to afford the title compound.

Example Synthesis of 2-(S)-{2-(R)-[2-(S)-Acetylam-ino-3-(4-hydroxy-phenyl)-propionylam-iino]-3acid [1-carbamoyl-2-(1H-indol-3-yI)ethyl]-amide

NH

2 OHNH I- NH 00 N H 1-f l-Carbamoyl-2-(lH-indol-3-yl)-ehylcarbamoyll-4-nitroguanidino-butylcarbamoyll-2- (R)-phenyl-eth lY-carbamic acid tert-bu~tyl ester To a solution of 2-(S)-(2-(R)-tert-butoxycarbonylanaino-3-phenyl-propionylamino)-5nitroguanidino-pentanoic acid (800mg, 1 7 mmoles), and 2-(S)-aniino-3-( lH-indol-3-yl)- WO 02/26774 WO 0226774PCT/USOI/30051 propionarnide (500mg, 1.2eq) in DMF (50mi) is added 1-hydroxybenzotriazole (350mg, 1-[3-(dimethylaminopropyl]-3-ethylcarbodiimide hydrochloride (491lig, 1 .5eq) and triethylamine l7nil, 5eq). The resulting solution is stirred at room temperature for twenty-four hours and then the solvents are removed under reduced pressure. The residue is partitioned between ethyl acetate (75ni1) and 10%1 sodium carbonate (75m1). The organics are dried over anhydrous magnesiumn sulfate, filtered and the solvents removed under reduced pressure. The crude product is purified by flash chormatography on silica gel (90:9:1 chloroform:methanol:anoniuin hydroxide) to afford the title compound.

2-(S)-(2-(R)-Arnino-3-phenyl-propionylamino)-5-nitroguanidinopentanoic acid rl-carbamoyl-2- (lH1-indol-3-yL)-etll-armide To a solution of l-carbamoyl-2-(lH-indol-3-yl)-ethylcarbamoyl]-4-nitroguanidinobutylcarbamoyl}-2-(R)-phenyl-ethyl)-carbamic acid tert-butyl ester (1.16g, 1.78mmoles) in methylene chloride (lO0rml) is added trifluoroacetic acid (50nil). The resulting solution is stirred at room temperature for twenty-four hours, and then the solvents are removed under reduced 1s pressure. The crude material is purified by reverse-phase ITPLC to afford the title compound.

2-0S-4 2-(R)-[2-(S)-Aetylamino-3-(4-hydroxyv-phenyl)-:propionla ol-3-ph Iypropionylamino 1-5-nitroguanidino-pentanoic acid F l-carbamovl-2-( 1H-indol-3-vl)-ethl3l-amide To a solution of 2-(S)-(2-(R)-arrino-3-phenyl-propionylamnino)-5-nitroguanidino-pentanoic acid [1-carbamoyl-2-( 1H-indol-3-yl)-ethyl]-arnide (250mg, O.378mmoles) and 2-(S)-acetylamino-3-(4hydroxy-phenyl)-propionic acid (100mg, l.2eq) in DMIF (50m1) is added 1-hydroxybenzotriazole (76mg, 1.5eq), 1-[3-(dimethylaminopropyl1-3-ethylcarbodiimide hydrochloride (107mg, 1 .Seq) and triethylamine (0.20m1, 4eq). The resulting solution is stirred at room temperature for twentyfour hours, and then the solvents are removed under reduced pressure. The crude material is purified by reverse-phase HPLC to afford the title compound.

2-(R)-r2-(S)-Acetvlarn-ino-3-(4-hydroxy-phenyl-propionylaniino-3-phenyacid [1-carbamcoyl-2-(1H-indol-3-vyl)-th l-mde To a solution of 2-(S)-{2-(R)-[2-(S)-acetylamino-3-(4-hydroxy-phenyl)-propionylamino]-3phenyl-propionylamnino }-5-nitroguanidino-pentanoic acid Ijl-carbamoyl-2-(lH-indol-3-yl)-ethyl]aruide (100mng, 0. 13rmoles) in methanol (50m1) is added acetic acid (51) and 5 palladium on abrium sulfate (70mg). The resulting suspension is hydrogenated at atmospheric pressure for twenty-four hours, filtered through celite and the solvents removed under reduced pressure. The crude product is purified by reverse-phase HPLC to afford the title compound.

WO 02/26774 WO 0226774PCT/US01/30051 Example 61 Synthesis of 2-(2-(R)-Aniino-3-phenyl-propionylamino)-5-(1-trityl-1H-iniidazol-4-yl)pentanoic acid [2-(S)-(1H-indol-3-yl)-1-methylcarbamoyl-ethiyl]-amide

NH

0

~NH

2-Anmino-5-(3H-imidazol-4-yl)-pentanoic acid methyl ester To a suspension of 2-tert-butoxycarbonylamino-5-(3H-imiidazol-4-yl)-pentanoic acid 1 1.Sinmoles) in methanol (6Onil) is added anhydrous hydrogen chloride until the solution is saturated. The solution is then heated to reflux for twenty-four hours and then cooled to room temperature. The solvents are removed under reduced pressure to afford the title compound.

2-(S)-(2-(R)-tert-Butoxycarbonylaniino-3-phenyl-propionylamino)-5-(3H-imidazol-4-yl) pentanoic acid metl ester To a solution of 2-(R)-(2-tert-butoxycarbonylamino-3-phenyt-propionic acid (500mg, l.88mmroles) and 2-aniino-5-(3H-iniidazol-4-yl)-pentanoic acid methyl ester (500mg, 1. leq is added hydroxybenzotriazoie (381mg,. 1 .5eq), 1 -I3-(dimethylamiinopropyl]-3-ethylcarbodiimide hydrochloride (540mng, 1 .5eq) and triethylamnine (1 .28m1, 5eq). The resulting solution is stirred at room temperature for twenty-four hours and then the solvents are removed under reduced pressure. The residue is partitioned between ethyl acetate (75m1) and 10% sodium carbonate (75m1). The organics are dried over anhydrous magnesium sulfate, filtered and the solvents removed under reduced pressure. The crude product is purified by flash chromatography on silica gel (90:9:1 chlaroform:methanol: ammonium hydroxide) to afford the title compound.

2-(S)-(2-(R)-tert-Butoxycarbovlanino-3-phenyl-uropionylamino)V5-(l-trityl-lH-imidazol-4-vD)pentanoic acid methyl este To a solution of 2-(S)-(2-(R)-tert-butoxycarbonaylainino-3-phenyl-propionylamino)-5-(3Himidazol-4-yl)-pentanoic acid methyl ester (300mg, 0.72mmnoles) in tetrahydrofuran (50aI) is added triphenylmethylchioride (220mg, 1.leq) and triethylamine 2eq). The resulting solution is stirred at room temperature for twenty-four hours and then the solvents are removed WO 02/26774 WO 0226774PCT/USOI/30051 under reduced pressure. The crode product is purified by flash chromatography on silica gel methanollchloroform) to afford the title compound.

2-(S)-(2-(R)-tert-Butoxycarbonylamino-3-phenyl-propionylarrnino)-5-( 1-trityl-lH-iDmidazol-4-yl)pentanoic acid To a solution of 2-(S)-(2-(R)-tert-butoxycarbonylamino-3-phenayl-propionylamino)-5-(1-trityl-1Hiniidazol-4-yl)-pentanoic acid methyl ester (300mg, 0.45rnmoles) in tetrahydrofuran (30m1) is added lithium hydroxide monohydrate (32mg, 1.2eq) and water (3m1) the resulting solution is stirred at room temperature for twenty-four hours, and then the solvents are removed under reduced pressure. The crude material is used directly in the next step.

1 l- l)-f -r2-(S%-u1-indol-3-yl)-l-methvlcarbamovl-ehv, rro1 -4-(1-trityl-TH-imidazol-4yl)-butylcarbamovll-2-nhenyl-ethyI I -carbamic acid tert-hutyl este To a solution of 2-(S)-(2-(R)-tert-butoxycarbonylamino-3-phenyl-propionylamino)-5-( 1-trityl-1Himidazol-4-yl)-pentanoic acid (293mg, 0.45rnmoles) and 2-(S)-arnino-3-( 1H-indol-3-yI)propionamide (1 17mg, 1.3 eq) is added PyBOP (301mg, 1 .3eq) and triethylamine 18m1, 3eq).

The resulting solution is stirred at room temperature for twenty-four hours, and then the solvents are removed under reduced pressure. The residue is partitioned between ethy acetate (75nI) and 19% sodium carbonate (75m1). The organics are dried over anhydrous magnesium sulfate, filtered, and the solvents removed under reduced pressure. The crude product is purified by flash chromatography on silica gel (5%o methanol/chloroform) to afford the title compound.

2-(2-(R)-Amino-3-:pheriyl-propionylamino)-5-( 1-trityL-111-imnidazo1-4-yl)-pentanoic acid P2-(S)- (lH-indol-3-yl)- t-methylcarbamoyl-ethyli-amide To a solution of I l-[2-(S)-(1H-Jndol-3-yl)-1-methylcabamoyl-ethylcarbamoyl -4-(1-trityllH-imidazol-4-yl)-butylcarbanoyl]-2-phenyl-ethyl}-carbamic acid tert-butyl ester (46 8mg, 0.,55 mmoles) in methylene chloride (32rn1) is added trifluoroacetic acid (l6nil). Triethylsilane is then added dropwise until the bright yellow color disappeared. The resulting solution is stirred at room temperature for twenty-four hours and then the solvents are removed under reduced pressure. The crude material is purified by reverse phase 1{PLC to afford the title compound.

Example 62 Synthesis of 2-(R)-[2-(S)-(2-Benzyl-6-phenyl-hexanoylaniino)-5-guanidinopentanoylamino]-3-naphtlialen-2-yl-propioriic acid methyl ester WO 02/26774 PCT/US01/30051 o 6-Phenyl-hexanoic acid methyl ester A solution of 6-phenyl-hexanoic acid (1.9g, 9.89mmoles) in methanol (50ml) is saturated with anhydrous hydrogen chloride and then heated to reflux for twenty-four hours. After cooling to room temperature, the solvents are removed under reduced pressure and the residue partitioned between chloroform and 10% sodium carbonate. The organics are dried over anhydrous magnesium sulfate, filtered and the solvent removed under reduced pressure to afford the title compound.

2-Benzvl-6-phenyl-hexanoic acid methyl ester To a cooled (-78 0 C) solution of 6-phenyl-hexanoic acid methyl ester (2.8g, 13.5mmoles) in anhydrous tetrahydrofuran (50ml) is slowly added a 2.0M solution of lithium diisopropylamide hexane/tetrahydrofuran (7.5ml, 1.leq). The resulting solution is stirred at -78oC for fifty minutes and then benzyl bromide (1.92ml, 1.2eq) is slowly added. The resulting solution is warmed to room temperature overnight and then the solvents are removed under reduced pressure. The residue is then partitioned between ethyl acetate and water. The organics are dried over anhydrous magnesium sulfate filtered and the solvents removed under reduced pressure. The crude product is purified by reverse-phase HPLC to afford the title compound.

2-Benzvl-6-phenyl-hexanoic acid To a solution of 2-benzyl-6-phenyl-hexanoic acid methyl ester (2.17g, 7.33mmoles), in tetrahydrofuran (100ml) is added lithium hydroxide monohydrate (880mg, 2eq) and water The resulting solution is heated to reflux for forty-eight hours and then cooled to room temperature. The solvents are removed under reduced pressure and the residue partitioned between ethyl acetate and 1M citric acid. The organics are dried over anhydrous magnesium sulfate, filtered and the solvents removed under reduced pressure. The crude product is purified by reverse phase HPLC to afford the title compound.

2-(S)-(2-Benzyl-6-phenyl-hexanoylamino)-5-nitroguanidino-pentanoic acid methyl ester To a solution of 2-benzyl-6-phenyl-hexanoic acid methyl ester (513mg, 1.82mmoles) and acid methyl ester (480mg, l.leq) in DMF (50ml) is added 1- WO 02/26774 WO 0226774PCT/USOI/30051 hydroxybeuzotriazole (319mg, 1 .3eq), 1-[3-(dimethylaniinopropyl]-3-ethylcarbodiimiide hydrochloride (45 1mg, 1.3eq) and triethylamine (0.74n1 3eq). The resulting solution is stirred at room temperatrure for twenty-four hours, and then the solvents are removed under reduced pressure. The crude material is purified by reverse-phase HPLC to afford the title compound.

2-(S)-(2-Benzyl-6-phienyl-hexanoylamino)-5-nitrofuanidino-:pentanoic acid To a solution of 2-(S)-(2-benzyl-6-phenyl-hexanoylamino)-5-nitroguanidino-pentanoic acid methyl ester (700mg, 1 .Srmoles) in tetrahydrofuran (40m1) is added lithium hydroxide monohydrate (180mg, 2eq), and water (3ml). The resulting solution is stirred at room temperature for twenty-four hours and then the solvents are removed under reduced pressure.

The crude product is purified by reverse-phase HIPLC to afford the title compound.

2-Aniino-3-naphthialen-2-:yl-:proionic acid methyl ester To a suspension of 2-arnino-3-napthtalen-2-yl-propionic acid (300mg, 1.39inmoles) in methanol (40m1) is added anhydrous hydrogen chloride until the solution is saturated. The resulting solution is heated to reflux for two hours and then cooled to room temperature, and thle solvents removed under reduced pressure to afford the title compound.

2-(R)-F2-(S)-(2-Benzyl-6-phenyl-hexanoylamino)-5-nitroguanidino-pntanvlariin1 -3naphthalen-2-yI-propionic acid methyl ester To a solution of 2-(S)-(2-benzyl-6-phenyl-hexanoylamino)-5-nitroguanidino-pentanoic acid (100mg, 0.22mmoles) and 2-amino-3-naphthalen-2-yl-propionic acid methyl ester (65mg, 1.leq) in DMI (30m1) is added 1-hydroxybenzotriazole (44mg, 1 eq), l-[3-(dimethylaminopropyl]-3ethylcarbodiimide hydrochloride (63mg, 1 .5eq) and triethyl amine (0.09m1 3eq). The resulting solution is stirred at room temperature for twenty-four hours and then the solvents are removed under reduced pressure. The crude material is purified by reverse-phase HPLC to affore the title compound.

2-(R-r2-S)-2-Benzyl-6pheni-exanolamino-5-ganidinopentanolaminoI -3-naphthalen-2yl-prgpionic acid methyl ester To a solution of 2-(R)-12-(S)-(2-benzyl-6-phenyl-hexanoylam-ino)-5-nitroguanidinopentanoylamino]-3-naphthalen-2-yl-propionic acid methyl ester (80mg, 0. 1 Iimoles) in methanol (30m1) is added acetic acid (3nil) and 5% palladium on barium sulfate (75mg). The resulting suspension is hydrogenated at atmospheric pressure for twenty-four hours, filtered through celite, and the solvents removed under reduced pressure. The crude product is purified by reverse-phase HPLC to afford the title compound as a mixture of four diastereomers.

61 WO 02/26774 WO 0226774PCT/USOI/30051 Example 63 Synthesis of 5-(1H-Imidazol-4-yl)-2-[3-phenyl-2-(R)-(3-phenyl-propionylamino)propionylamino]-pentanoic acid (1-(S)-methylearbamoyl-2-naphthalen-2-yl-ethyl)-amide N N

N

.NJ

3-Phenyl-2-(R)-(3-phenyl-propiovlamino)-:propionic acid methlyl ester To a solution of 3-phenyl-propionic acid (1-0g, 6.66mmoles) and 2-(R)-amino-3-phenyl-propionic acid methyl ester 19g, leq) in DMIF (60m1) is added 1-hydroxybenzatriazole (1.35g, l.5eq), 1r3-(dimethylaminopropyl] -3-ethylcarbodimride hydrochloride (1 .90g, 1 .5eq) and triethylamine (2.7m1, 3eq). The resulting solution is stirred at room temperature for twenty-four hours, and then the solvents are removed under reduced pressure. The residue is partitioned between water and ethyl acetate, dried over anhydrous magnesium sulfate, filtered and the solvents removed under reduced pressure to afford the title compound 3-Phenyl-2-(R)-(3-phe~nyl-propionylaminio)-propionic acid To a Solution of 3-phenyl-2-(R)-(3-phenyl-propionylamino)-propioriic acid methyl ester 4.82mmoles) in tetrahydrofuran (70m1) is added lithium hydroxide monohydrate (434mg, and water (5rn1). The resulting solution is stirred at room temperature for twenty-four hours, and then the solvents are removed under reduced pressure. The crude material is purified by reversephase HPLC to afford the title compound.

5-(311-lnidazol-4-yl)-2-[3-:phenyl-2-(R)-(3-:phenyl-propionylan-inro)-propionvlarmmol pentanoic acid methyl ester To a solution of 3-phenyl-2-(R)-(3-phenyl-propionylamino)-propionic acid (376mg, 1 .26nunoles) and 2-ainino-5-(3H-iidazol-4-yl)-pentanoic acid mnethyl ester (355mg, 1. leq) in DMF (50mi) is added l-hydroxybenzotriazole (256mg, 1.5eq), l-13-(diiethylamainopropylj-3-ethylcarbodiimide hydrochloride (362mg, 1 .5eq) and trietbylamine (O.863m1, 5eq). The resulting solution is stirred at room temperature for twenty-four hours and then the solvents are removed under reduced pressure. The crude product is purified by reverse-phase FFPLC to afford the title compound.

24f3:-?hen 1-2-(R)-(3-phenyl-:propiolamino)-propionylanmino-5-(l-tritl-IH-nidazol-4-vlDpentanoic acid meathyl ester WO 02/26774 WO 0226774PCT/USOI/30051 To a solution of 5-(3H-iinidazol-4-yl)-2-[3-pheniyl-2-(R)-(3-phenyl-propionylamino)propionylam-ino]-pentanoic acid methyl ester (473mg, 1.O2mmoles) in tetrahydrofuran (50m1) is added triphenylmethyichloride (34 1mg, 1.2eq), and triethylamine (O.42m1, 3eq). The resulting solution is stirred at room temperature for twenty-four hours and then the solvents are removed under reduced pressure. The crude product is purified by flash chromatography on silica gel (5 methanollchloroform) to afford the title compound.

2F31Phen 1-2-(R)-(3-phenyl-:propionvylanmino)-propi nlaminol-5-(1-triyl-lH-irnidazol-4yl)pentanoic acid To a solution of 2-[3-phenyl-2-(R)-(3-phenyl-propionylamino)-propionylamino]-5-(1-trityl-1Himidazol-4-yl)-pentanoic acid methyl ester (380mg, O.54mmoles) in tetrahydrofuran (8Onil) is added lithium hydroxide monohydrate (48mg, 1 .5eq) and water (l0ml). The resulting solution is stirred at room temperature for twenty-four hours, and then the solvents are removed under reduced pressure. The crude material is used directly in the next step.

(1 -(S)-Methylcarbamovl-2-naphthalen-2-vl-ethvl)-carbamic acid tert-butyl ester To a solution of 2-(S)-tert-butoxycarbonylamino-3-naphthalen-2-yl-propionic acid 4.76mmoles) in DMF (50m1A) is added methyl amine (3.0ml of a 2.OM solution in tetrahydrofuran), PyBOP (3.7g, 1.5eq), and triethylamine (1.95m1, 3eq). The resulting solution is stirred at room temperature for twenty-four hours, and then the solvents are removed under reduced pressure. The residue is partitioned between ethyl acetate and 10% sodium carbonate, the organics are dried over anhydrous magnesium sulfate,filtered and the solvents removed under reduced pressure. The crude product is purified by flash chromatography on silica gel methanol/chloroform) to afford the title compound.

2-(S)-Amfino-N-methyl-3-nanhthalen-2- yl-oropionamide To a solution of (1-(S)-methylcarbamoyl-2-naphthalen-2-yl-ethyl)-carbanic acid tert-butyl ester 3.96mmoles) in methylene chloride (lO0mi) is added trifluoroacetic acid (50rmi). The resulting solution is stirred at room temperature for twenty-four hours, and then the solvents are removed under reduced pressure. The crude material is purified by reverse-phase HIPLC to afford of the title compound.

2-(S)-[3:Phen l-2-(R)-(3-phenyl-propionvlamino)-propionylaniinol-5-(1-trityl-lH-imidazo1-4-vl)pentanoic acid (1-(S)-met~hylcarbamoyl-2-nanhthlen-2-vyl-ethyl)-anmide To a solution of 2-13-phenyl-2-(R)-(3-phenyl-propionylarmino)-propionylainino]-5-( 1-trityl-1Himidazol-4-yl)-pentanoic acid (383mg, O.S3mr-noles) in DMIP (50m1d) is added 2-(S)-arnino-N- WO 02/26774 WO 0226774PCT/USOI/30051 methyl-3-naphthalen-2-yl-propionamide (147mg, 0. 8eq), PyBOP (420mg, 1 .5eq) and triethyl am-ine (0.220m1, 3eq). The resulting solution is stirred at room temperature for forty-eight hours, and then the solvents are removed under reduced pressure. The residue is partitioned between ethyl acetate and 10%1 sodium carbonate, the organics are dried over anhydrous magnesium sulfate, filtered and the solvents removed under reduced pressure. The crude product is purified by flash chromatography on silica gel methanollchloroform) to afford the title compound.

lH-linidazol-4-yl)-2-F3:-henyl-2-(R)-(3-vphenyl-propionaylamino)-:propionvlamino] -entanoic acid (1 -(S)-methylcarbamoyl-2--naphfthalen-2-yl-ethyl)-amide To a solution of trifinoroacetic acid (20m1) in methylene chloride (40m1) is added phenyl-2-(R)-(3-phenyl-propionylamino)-propionylamino--5-(1-trityl-H-iuidazol-4-yl)pentanoic acid (1-metliylcarbamoyl-2-naphthlen-2-yl-ethyl)-amide (44 1mg, 0.48 9mmoles).

Triethylsilane: is then added dropwise until the bright yellow color just disappeared. The resulting solution is stirred at room temperature for twenty-four hours and then the solvents are removed under reduced pressure. The crude product is purified by flash chromatography on silica gel (90:9:1 chloroform:methanol: ammonium hydroxide) to afford the title compound as a mixture of diastereomers. The diastereomers are separated by reverse-phase UPLC to afford an earlier eluting diastereomer and a later eluting diastereomer.

Example 64 Synthesis of 2-(S)-{2-(R)-[2-(S)-Acetylamino-3-(4-hydroxy-phenyl)-propionylaniino]-3phenyl-propionylaniino}-5-guanidino-pentanoic acid (1-(S)-methylcarhamoyl-2naphthalen-2-yl-ethyl)-amide 0 0 0 1 -(R)-[4-Nitro~yuanidino- l-CS)-(1-(S)-methvlcarbamoyl-2-naphthalen-2-vl-thylcarbamoyL)butvlearbaloyll -2-phe nvi-ethyI I -carbamic acid tert-butyl ester To a solution of 2-(S)-(2-(R)-tert-butoxycarhonylam-ino-3-phenyl-propioniylaniino)-5nitroguanidino-pentanoic acid (500mg, 1 .07mmoles) and 2-(S)-amino-N-methyl-3--naphthalen-2yl-propionainido (440mg, 1.2eq), is added PyBOP (836mg, 1.5eq) and triethylamnine (0.58m1, 4eq). The resulting solution is stirred at room temperature overnight and then the solvents are WO 02/26774 WO 0226774PCT/USOI/30051 removed under reduced pressure. The residue is partitioned between ethyl acetate and sodium carbonate, dried over anhydrous magnesium sulfate, filtered and the solvents removed under reduced pressure. The crude product is purified by flash chromatography on silica gel (90:9: 1 chloroform:methanol: ammonium hydroxide) to afford the title compound.

2-(S)-(2-(R)-Arnino-3-:phenyl-~propignylamino)-5-nitroguanidino-pentanoic acid methylcarbamoyl-2-nahthalen-2-vyl-ethyl)-amide To a solution of I 1 -(R)-[4-nitroguanidino-1-(S)-( 1-(S)-methylcarbamoyl-2-naphthalen-2-ylethylcarbamoyl)-butylcarbamoyl] -2-phenyl-ethyl }-carbamic acid tert-butyl ester (500mg, 0.74mmoles) in methylene chloride (6Ond) is added trifluoroacetic acid (30m1). The resulting solution is stirred at room temperature for twenty-four hours, and then the solvents are removed under reduced pressure. The crude product is purified by reverse-phase HPLC to afford the title compound.

2-(R)-r2-(S)-Acetylamino-3-(4-.hydxy-phen1)-pr oiylaminol -3-Phenypropioniylainiino I-5-nitroguaniidinto-p2entanioic acid (1-(S)-i-ethylcarbamioyl-2-naiphthaen-2-ylethy1 -amnide To a solution of 2-(S)-(2-(R)-aniino-3-phenyl-propionylamnino)-5-nitroguanidino-pentanoic acid (1-(S)-methylcarbamoyl-2-naphthalen-2-yl-ethyl)-amide (400mg, 0.58mmoles) in DMF (6Ornl) is added 2-(S)-acetylamino-3-(4-hydroxy-ph-enyl)-propionic acid (155mg, 1 .2eq), PyBOP (410mg, l.2eq), and triethylarnine (0,32m1, 4eq). The resulting solution is stirred at room temperature for twenty-four hours, and then the solvents are removed under reduced pressure. The residue is partitioned between ethyl acetate and 10% sodium carbonate, the organics are dried over anhydrous magnesium sulfate, filtered and the solvents removed under reduced pressure. The crude product is purified by flash chromatography on silica gel (90:9:1 chloroform: methanol:ainmonium hydroxide) to afford the title compound.

2-(R)-[2-(S)-Acetvlaniino-3-(4-hydroxy-phe nyl-roi la inol-3phnpronionvlamino 1-5-guanidino-pentanoic acid (1-(S)-methvlcarbamoyl-2-nahthalen-2-yl-ethyl)amide To a solution of 2-(R)-[2-(S)-acetylamino-3-(4-hydroxy-phenyl)-propionylamino]-3phenyl-propionylamino }-5-nitroguanidino-pentanoic acid (l-(S)-methylcarbamoyl-2-naphthalen- 2-yl-ethyl)-amide (330mg, 0.42mmoles) in methanol (50m1) is added acetic acid (51) and 501 palladium on barium sulfate (325mg). The resulting suspension is hydrogenated at atmospheric pressure for twenty-four hours, and then filtered through ce-lite. The solvents are removed WO 02/26774 WO 0226774PCT/USOI/30051 reduced pressure and the crude product purified by reverse-phase NPLC to afford the -title compound.

Example Synthesis of 5-Guanidino-2-(S)-{2-(R)-[3-(4-hydroxy-phenyl)-propionylamino]-3-phnylpropionylarmino}-pentanoic acid (1-(S)-methylcarbamoyl- 2-naphthalen-2-yI-ethyl)-amide N

"'Y

To a solution of 2-(S)-(2-(R)-amiino-3-phenyl-propionylamino)-5-nitroguanidino-pentanoic acid I0 (l-(S)-methylcarbamoyl-2-naphthalen-2-yl-ethyl)-aniide (268mng, 3.88nimoles) and 3-(4-hydroxyphenyl)-propionic acid (77mg, 1.2eq) is added 1-13-(dimetlylaminopropylj-3-ethylcarbodiiniide hydrochloride (111mg, 1.5eq), l-hydroxybenzotriazole (78 mg, 1.5 eq) and triethylamine (0.21 nml, 4 eq). The resulting solution is stirred at room temperature for twenty-four hours and then the solvents are removed under reduced pressure. The crude material is purified by reverse-phase HPLC to afford the title compound.

5-Guanidino-2-(S)-I 2-(R)-r3-(4-hydroxy-phe~nyl)-prop~ionylamninol-3-phecnyl-propionvlaminlo I1pentanoic acid (1-(S)-methylcarbamoyl-2-naphthalen-2-yl-ethyl)-amide.

To a suspension of 5-nitroguanidino-2--(S)--{2-(R)-[3-(4-hydroxy-phenyl)-propionylamino] -3phenyl-propionylaminol-pentanoic acid (1-(S)-methylcarbamoyl-2-naphthalen-2-yl-ethyl)-amide (141mg, 0. l9nnnoles), and 5% palladium on barium sulfate (100mg) in methanol (45ni1) is added acetic acid (5m1). The resulting suspension is hydrogenated at atmospheric pressure four twentyfour hours, filtered through celite, and the solvents are removed under reduced pressure. The crude product is purified by reverse-phase HPLC to afford the title compound.

Example 66 Synthesis of 5-Guanidino-2-(S)-113-phenyl-2-(R)-(2-phenyl-ethanesulfonylamino)propionylaminol-pentanoic acid (1-(S)-nethylcarhamoyl-2-naphthalen-2-yL-ethyl)-anide WO 02/26774 PCT/US01/30051 2-Phenyl-ethanesulfonyl chloride To a biphasic solution of 2-phenyl-ethanethiol (10.0g, 72.5mmoles) in 100mi of ice water is added acetic acid (20ml). This solution is then saturated with chlorine gas for five minutes. The aqueous solution is then extracted with ethyl ether, dried over anhydrous magnesium sulfate, filtered and the solvents removed under reduced pressure to afford the title compound.

3-Phenvl-2-(R-(2-phenyl-ethanesulfonvlamino)-propionic acid methyl ester To a solution of 2-amino-3-phenyl-propionic acid methyl ester (700mg, 3.9mmoles) in tetrahydrofuran (30ml) is added dropwise 2-phenyl-ethanesulfonyl chloride (1.2g, 5.88mmoles).

To this solution is added triethylamine (1.55ml, 3eq). The resulting solution is stirred at room temperature for twenty-four hours and then the solvents are removed under reduced pressure.

The crude product is purified by reverse-phase HPLC to afford the title compound.

3-Phenyl-2-(R)-(2-phenyl-ethanesulfonylamino)-propionic acid To a solution of 3-phenyl-2-(R)-(2-phenyl-ethanesulfonylamino)-propionic acid methyl ester (890mg, 2.56mmoles) in tetrahydrofuran (50ml) is added lithium hydroxide monohydrate (260mg, 1.5eq) and water (5ml). The resulting solution is stirred at room temperature for twentyfour hours and then the solvents are removed under reduced pressure. The crude product is purified by reverse-phase HPLC to afford the title compound.

[4-Nitroguanidino-1-(R)-(1-(S)-methylcarbamovl-2-naphthalen-2-vl-ethlcarbamovl)-butv11carbamic acid tert-butvyl ester To a solution of 2-amino-N-methyl-3-naphthaolen-2-yl-propionanide (1.0g, 2.92mmoles) and 2acid (1.12g, 1.2eq) is added 1-[3- (dimethylaminopropyl]-3-ethylcarbodiimide hydrochloride (837mg, 1.5eq), 1hydroxybenzotriazole (592mg, 1.5eq), and triethylamnine (1.2ml, 3eq). The resulting suspension is stirred at room temperature for twenty-four hours and then the solvents are removed under reduced pressure. The residue is partitioned between ethyl acetate and 10% sodium carbonate, the organics are dried over anhydrous magnesium sulfate, filtered, and the solvents removed WO 02/26774 WO 0226774PCT/USOI/30051 under reduced pressure. The crude product is purified by reverse-phase HPLC to afford the title compound.

acid (l-(S)-rnethylcarbamov-2-nahthalen-2-yl-ethylD-ande To a solution of [4-niitroguaniidino-l-(R)-(l-(S)-methylcarbamoyl-2-naphtlhalen-2-ylethylcarbamoyl)-butylj-carbamic acid tert-butyl ester (800mg, l.52mmoles) in methylene chloride (4Ornl) is added trifluoroacetic acid (20m1A). The resutting solution is stirred at room temperature for twenty-four hours and then the solvents are removed under reduced pressure.

The crude product is purified by reverse-phase HPLC to afford the title compound as the trifluoroacetic acid salt.

5-Nitroguanidino-2-(S)-[3-phenly-2-(RY-(2-phenvyl-ethanesulfonvlamimo)-propionvlaninopentanoic acid (1-(S)-methylcarbamoyl-2-naphthalen-2-vyl-ethvl)-amide To a solution of 3-phenyl-2-(R)-(2-phenyl-ethanesulfonylamino)-propionic acid (150mg, 0.45rnoles) and 2-(R)-amrino-5-guanidino-pentanoic acid (1 -(S)-methylcarbamoyl-2-naphthalen- 2-yl-ethyl)-anride (270mng,1. leq) is added 1-[3-(dir-netliylamiinopropyl]-3-ethiylcarbodiilide hydrochloride (130mg, 1.5eq), l-hydroxybenzotriazole (9 1mg, 1.5eq), and triethyl amine (0.25m1, 4eq). The resulting solution is stirred at room temperature for twenty-four hours and then the solvents are removed under reduced pressure. The crude product is purified by preparative HPLC to afford the title compound.

5-Guanidino-2-(S)-13-phenyl-2-(R)-(2-phenyl-ethanesulfonylaino)-:propionlarminolpetanoic acid (1-(S)-mehlerbamoyl-2-nphthalen-2-y-ehyl)-amide To a solution of 5-nitroguanidino-2-(S)-!13-phenyl-2-(R)-(2-phenyl-ethanesulfonylamino)propionylamninol-pentanoic acid (l-(S)-methylcarbamoyl-2-niaphthalen-2-yl-ethyl)-arnide (23 0mg, 0.3 immoles) in methanol (45m1) is added acetic acid (5m1) and 5% palladium on barium sulfate (200mg). The resulting suspension is hydrogenated at atmospheric pressure fur twenty-four hours, and then filtered through celite. The solvents are removed under reduced pressure and the crude product purified by reverse-phase HIPLC to afford the title compound as the trifluoroacetic acid salt.

Example 67 Synthesis of Ac-(carba-Ft)-RW-N1 2 (11) This compound is prepared according to the plans presented in Schemes IA&B: WO 02/26774 WO 0226774PCT/US01/30051 0 C' o OH oo~ DMAP BON o aBS 4

E.

1 2 J %Xylenes BGHo2 1) NaHMDS H O0. 2) 8 0 cv 3a R H Sb R B oo 4 Scheme 1A [1-(4-Chloro-benz'l)3-(2,2-dimethl-4,6-dioxo-f1 .3ldioxan-5-yl)-3-oxo-propyll -carbamic acid tert-butyl ester (1) To a well-stirred mixture of Boc-(S)-3-amino-4-(4-chlorophenyl)-butyric acid (5.0 g, 16 mmaol), 2,2-dimethyl-1,3-dioxane-4,6-dionie (2.54 g, 17.6 mmol) and DMIAP (24 mmol) in DCM (160 ml) at OTC is added EDCI (24 nunol) in one portion. The resulting mixture is stirred at 0 0 C for 1 h, then at room temperature for 18 h. DCM (100 nil) is added and the mixture is washed with water (2 x 50 ml), 5% aqueous potassium hydrogen sulfate (3 x 50 ml), 5% aqueous sodium bicarbonate (1 x 50 ml), and brine (1 x 50 ml). The organic layer is dried over anhydrous magnesium sulfate and concentrated to yield 1.

1l-(4-Chloro-benzyl)3-(2.2-dimethyl-4,6-dioxo-[1 ,3]dioxan-5-yl)-:propyll-caxban-ic acid tert-butvl ester (2) Sodium borohydride (63.8 mmol, 4.0 eq.) is added portionwise over 1 h to a well-stirred solution of 1 15.96 mmol) in a mixture of DCM (180 ml) and acetic acid (10 ml, 175 mmol, 11.0 eq.) at 0' C. The resulting mixture is stirred at 0' C for 1 h and then at room temperature for 64 h. The reaction mixture is diluted with DCM (150 nil) and washed with water (I x 50 nil) and brine (2 x ml). The organic layer is dried over anhydrous magnesium sulfate and concentrated by rotary evaporation to yield 2.

6-(4-Chloro-benzyl)-piperidin-2-one (3a) and 2-(4-Chloro-benzvl)-6-oxo-piperidine-l-carbox(Ylic acid tert-butLv ester (3b) A stirred mixture of 2 (15.32 mmol) and xylene (140 ml) is heated at reflux for 6 h. The solvent is removed by evaporation at 370 C in vacuo to yield crude 3a. This material is combined with di-tert-butyl dicarbonate (5.0 eq.) and DIAP (0.3 eq.) in DCM (100 ml) and the mixture is WO 02/26774 PCT/US01/30051 stirred at room temperature for 40 h. The solvent is removed by rotary evaporation and the residue is purified by flash chromatography on silica gel using EtOAc-hexane (1:19, 500 ml and 1:9, 1300 ml) as an eluant to yield 3b.

3-Benzvl-6-(4-Chloro-benzyl)-2-oxo-piperidine--carboxylic acid tert-butyl ester (4) Sodium bis(trimethylsilyl)amide (5.16 ml of a 1.0 M solution in THF, 1.0 eq.) is added dropwise to a well-stirred solution of 3b (1.67 g, 5.16 mmol) in a mixture of THF-DMIE 100 ml) at 78°C under argon and the resulting mixture is stirred for 0.5 h more at -78 0 C. To this stirred, cold mixture is added a solution of benzyl bromide (0.882 g, 5.16 mmol, 1.0 eq.) in THF (5 ml) and stirring is continued at -78 0 C under argon for 2 h. The reaction mixture is quenched with a saturated aqueous solution of ammonium chloride (20 ml) and stirring is continued for 10 min.

more. The mixture is then partitioned between DCM (80 ml) and water (40 ml) and the water phase is extracted with DCM (2 x 40 ml). The combined organic phase is washed with water (1 x ml), dried over anhydrous magnesium sulfate, and concentrated to give crude 4, which is purified by flash column chromatography on silica gel using EtOAc-hexane (1:39, 1000 ml and 1:19, 1300 ml) as an eluant to afford 4 (single diastereomer).

2-(R)-Benzyl-5-(R)-tert-butoxycarbonylamino-6-(4-chloro-phenyl)-hexanoic acid ('Boc-(carba-4- Cl-Ff)-OH', To a well-stirred solution of 4 (1.152 g, 2.783 mmol) in a mixture of THF-water are added lithium hydroxide monohydrate (0.467 g, 11.132 mmol, 4 eq.) in one portion and 35% hydrogen peroxide (1.95 ml, 8.0 eq.) by dropwise addition. The resulting mixture is stirred at 0 C for 1 h and then at room temperature for 16 h. The reaction mixture is acidified with aqueous hydrochloric acid (11.5 ml, IN) at 0°C and is extracted with DCM (4 x 40 ml). The organic layer is dried over anhydrous magnesium sulfate, concentrated, and the residue triturated to yield (single diastereomer).

WO 02/26774 PCT/USOI/30051 NO S H H 'J0 2 iH~jl N- N0 2 rN YN -NO NH I\NH NH 2

NH

2

I"

OH H'e\ H I )TFA, DOM p-ISA H BoN N Non NN H oEDOI, I-O~t, NMM, DMF N 2) pTSA H NH 9H2N 0 )-N 6 7 6oc-(carba-4-GI-f)-OH NMM, DMFt ci 2

I)TFA,DCMN

S.\H 2) p'ABoNH p-TPSA H o 2pSAH

H

ACH

rDOI, HOSt NM," IDMF ci 14 N0H 2 Y

H

2 P a O I NH N "'2 H H H NH

HN

2 N 0 HN _N Scheme lB Boc-R(NO2)-W-NH 9 (6) HOBt (1.0 eq.) is added to a well-stirred solution of Boc-R(N0 2 )-OH (0.639 g, 2.0 mmol) and B-

W-NH

2 HC1 (0.479 g, 2.0 rnmol) in DMF (10 ml) at OTC followed by addition of EDGI (1.1 eq.) and NMM (0.484 nil, 2.2 The resulting mixture is stirred at OTC for 1 h and then at room temperature for 4-18 h. The reaction mixture is diluted with EtOAc (100 ml) and is washed with water (1 x 20 ml), 1 N aqueous hydrochloric acid (2 x 10 ml), saturated aqueous sodium bicarbonate (2 x 10 ml), and brine (1 x 10 ml). The organic layer is dried over anhydrous magnesium sulfate and concentrated to yield 6.

P-TSA H-R(N',O -W-NIH?,(7) A solution of 6 (0.697 g, 1.381 minol) in a mixture of TFA-DCM-water (10:40:0.5, 15 ml) is stirred at room temperature for 2-4 h. p-Toluenesulfonic acid, monohydrate (1.0 eq.) is added to the reaction mixture. After being stirred for 10 m the mixture is concentrated by evaporation in vacuc and the residue is triturated with ether-hexane 1) to yield 2.

Boc-(carba-4-Cl-Ff)-R(NO 2 ,)-W-NH2 (8 The procedure of preparing compound 6 is followed and 7 (0.5 mmol) is coupled with Boc- (carba-4-Cl-Ff)-OH 0.5 mmol, 1.0 eq.) to yield 8.

P-TSA 9 H-(carba-4-Cl-Ff)-R(NO)-WNIT (9) The method of preparing compound 7 is followed and 9 is obtained from 8.

71 WO 02/26774 PCT/USOI/30051 Ac-(carba-4-Cl-Ff)-R(N&)-W-NH1 2 The method of preparing compound 6 is followed and 9 (0.472 mmol) is coupled with acetic acid to yield crude 10 which is purified by preparative HPLC (C, 8 to afford Ac-(carba-Ff)-R-W-NM2 (11) A mixture of 10 (200 mg, 0.263 mmol), and 300 rug of 5% Pd-BaSO 4 (unlreduced) in MeOH- HOAc (10: 1, 22 nil) is hydrogenated under 42 psi of hydrogen at room temperature for 17 h. The catalyst is removed by filtration through a pad of Celite, which is washed with MeOH. The filtrate is concentrated to give crude 11 which is purified by preparative HIPLC (C 18 and the product is further triturated with ether to yield 11 as a TFA salt.

Examples 68 and 69 Syntheses of 3-(4-hydroxyphenyl)propanoyl-Atc-R-tryptainde (17) and 3-phenylpropanoyl-Atc-R-tryptamide (19) These examples are prepared according to the plan presented in Scheme 11: H" H H N0 N YN N02 N N- 0

H

2

H

2 N NH 2 NH Boo., OH PH TFA H H a800, HO~t, DMF TFA, DCM NHN 0 H 0ho N N*2 NN0 2 12 1

~NH

2 HFo-Atc-OH C(A N OHEDC.,

HOAI,

NH H 0 NH 0TEA 18 801 HO~t, DMF ~NN NH 'O H N NH, H~fPd-BaS0 4 I 9Q("N1

HNH

2 H a NH DEA,ODMF NMdN N NH 15 14 0 H NH 2

HOH

CN 001I, HOBt, DMF NHH 0 NH

HH

aN N -N2N NH -19

Y~O

NH

2 Hd NHBS 0 NH NHH 0 NH NH 0 16 -17 Scheme 11 Boe-R(NO,))-tryptamide (12) HOBt (1.0 eq.) and EDCI (0.949 g, 4.95 nimol, 1.1 eq.) are added sequentially to a well-stirred mixture of Boc-R(N0 2 )-OH (1.437 g, 4.5 marnol) and tryptamine (0.721 g, 4.5 minol) in DMIF 72 WO 02/26774 PCT/US01/30051 ml) at o0C. The resulting mixture is stirred for 30 minutes at 0°C and then at room temperature for -20 h. The mixture is diluted with EtOAc (100 ml) and is then washed successively with water (2 x 20 ml), 5 aqueous citric acid (3 x 10 ml), 5% aqueous sodium bicarbonate (2 x ml), and brine (2 x 20 ml). The organic layer is dried over anhydrous sodium sulfate and concentrated by rotary evaporation to yield 12, which is used directly in the next step.

H-R(N0 2 )-trvptamide (13) A solution of 12 (1.985 g, 4.3 mmol) in a mixture of trifluoroacetic acid-dichloromethane-water (10:40:0.5, 50 ml) is stirred at 0°C for 15 minutes and then at room temperature for 16 h. The solvent is removed by rotary evaporation and the residue is co-evaporated with methanol (3 x ml). The crude product is then purified by preparative HPLC to yield 13.

Fmoc-Atc-R(NO)-tryptamide (14) HOAt (0.409 g, 3.005 mmol), EDCI (0.632 g, 3.306 mmol, 1.1 and TEA (l.leq.) are added sequentially to a well-stirred solution of 13 (1.086 g, 3.005 rmnol) and Fmoc-Atc-OH (1.242 g, 3.005 mmol) in DMF (15 ml) at 0°C. The resulting mixture is stirred for 30 minutes at o0C and then at room temperature for 21 h. Work-up as described above for 12 gives crude 14. This material is used directly in the next step without further purification.

H-Atc-R(NOz)-trvptamide A solution of 14 (2.27 g, 3.0 mmol) in a mixture of DEA-DMF 30 ml) is stirred at room temperature for 3 h. The solvent is removed by evaporation under reduced pressure and the residue is triturated with ether-hexane 100 ml) to yield 3-(4-hydroxyphenvl)propanoyl-Atc-R(NOg)-tryptamide (16) The procedure of preparing 12 is followed and 0.603 g of crude 16 is obtained from 15 (0.535 g, mmol) and 3-(4-hydroxyphenyl)propanoic acid (0.166 g, 1.0 mmol). This crude product is purified by preparative HPLC to yield 16.

3-(4-hydroxyphenyl)propanoyl-Atc-R-tryptamide (17) A mixture of 16 (0.270 g, 0.405 mmol) and 5% Pd-BaSO 4 (unreduced, 0.270 g) in MeOH-HOAc (10:1, 22 ml) is hydrogenated at room temperature and atmospheric pressure for 17 h. The catalyst is removed by filtration through a pad of celite and the filtrate is concentrated by rotary evaporation. The residue is purified by preparative HPLC and triturated with ether to yield 17.

3-phenylpropanovl-Atc-R(NO2)-trvptamide (18) WO 02/26774 PCT/USOI/30051 The procedure of preparing 16 is followed and 18 is obtained from 0.535 g (1.0 mind) of 15 and 3-phenyipropainoic acid 150 g, 1.0 mmol).

3-phen~ylpropanoyl-Atc-R-taptaniide (19) The procedure of preparing 17 is followed and 19 is obtained as a hydrated TFA salt from 0.270 mg ofl18.

Example Synthesis of 3-Phenylpropanoyl-f-N(Me)R-tryptamide This example is prepared according to the plan presented in Scheme Ill: NNH

H

I NyNNNNN ~Tco H 2 N HNL I N 2 NNH B H TO I 1) TFA, DGM P-TSA 0 f H Boo,. HBe N I aEDCI, H0Bt, DIVF 2)NHS HN N NH NHN I H20 21 9 0 NTos IBoG-f H ".jNPH IPyorop DCM NH 1 0 ~NH 0IE \ETA, Dmr H At 240 HNr To I 0 HN, Tos 9 H

H

0 IE Ua~q. NH 3 r N TrA DCIM N N H NH I:PN310_CNH SONH 1~ 0 N 0 NH 23 22 NH 0 ~NH Scheme III Boc-N(Me)-R(Tos)-tryptamide The procedure of preparing 12 (Scheme I) is followed and crude 20 is obtained from 1.991 g nimol) of Boc-N(Me)-R(Tos)-OH and 0.721 g (4.5 nimol) of tryptamine. This crude material is used in the next step without further purification.

P-TSA H-N(Me)-R(Tos)-tryptamide (21) The procedure of preparing 7 (Scheme 0) is followed and 21 is obtained as a p-TSA salt from 2.63 1 g (4.5 =iol) of Boc-f-N(Me)-R(Tos)-tryptamide (22) DIEA (0.3 10 nil, 1.772 minol, 2.0 eq.) is added to a well-stirred maixture of 21 (0.4293 g, 0.886 nimol), Boc-f-OH (0.235 1 g, 0.886 nimol), and PyBrop (0.3795 g, 0.886 inmol) in DCM (10 al) at 0 0 C. The resulting reaction mixture is stirred at room temperature for 16 h and then is diluted with EtOAc (50 ml). The organic layer is washed successively with water (1 x 10 ml), 5%1o WO 02/26774 PCT/US01/30051 aqueous sodium bicarbonate (2 x 10 ml), and brine 2 x 10 ml) and is dried over anhydrous sodium sulfate. Removal of the dessicant and evaporation of the volatiles under reduced pressure yields crude 22.

H-f-N(Me)-R(Tos)-tryptamide (23) A solution of 22 (0.609 g, 0.833 mmol) dissolved in a mixture of TFA-DCM-water (10:40:0.5, ml) is stirred at 0 C for 1 h, and then is kept in a refrigerator at 4°C for 72 h. The solvent is removed by evaporation in vacuo and the residue is then twice co-evaporated with DCM (10 ml The crude material so obtained is purified by preparative HPLC and triturated with ether to yield 23.

3-Phenvlpropanovl-f-N(Me)-R(Tos)-tryptamide (24) TEA (0.209 mmol, 1.1 eq.) is added to a well-stirred mixture of 23 (0.120 g, 0.19 mmol), 3phenylpropanoic acid (0.19 mmol), and EDCI (0.209 mmol, 1.1. eq) in DMF (3 ml) at 0°C. The resulting mixture is stirred at 0°C for 0.5 h and then at room temperature for 16 h. The reaction mixture is diluted with EtOAc (30 ml) and is washed successively with water (2 x 5 ml), aqueous citric acid (3 x 3 ml), 5% aqueous sodium bicarbonate (2x 3 ml), and brine (2 x 5 ml).

The organic layer is dried over anhydrous sodium sulfate, concentrated, and then triturated with ether to yield crude 24.

3-Phenylpropanoyl-f-N(Me)-R-tryptamide Compound 24 (100 mg, 0.13 mmol) is dissolved in liquid ammonia (25 ml) at -78 0 C and small pieces of metallic sodium are added with vigorous magnetic stirring until a blue color persists for 2 h. The excess reagent is destroyed with ammonium acetate and the ammonia is removed by evaporation at room temperature. The residue is dissolved in methanol and the solution is filtered through a pad of silica gel that is subsequently washed with more methanol. The combined methanol filtrates are concentrated by rotary evaporation to give a crude material which is purified by preparative HPLC to yield 25 as a hydrated, TFA salt.

Examples 71 through Syntheses of H-YfRW-NH2(32), Bc-YfRW-NH2 CH 3 (CH)sCO-YfRW-NH 2 Bc- YfRW-tryptamide and CH 3

(CH

2 )sCO-YfRW-tryptamide (42) These examples are prepared according to the plan presented in Scheme IV: WO 02/26774 PCT/US01/30051

+NH

2 EDGI, HOBt HN W~e TEA, DMF

H

N N 02 1) TFA, DCM 0cj NHz 2) p-TSA OY_ MU p-S~0 27 Boo-V-OH EDGI, H-OMt TEA, DMF

TFA

Scheme IV 33RH= (CH2)2CH-1 35RH= (CH2)aGH3 39 R =(CH 2 2

CH

3 41RH= (CH 2 ).Co

SH

2 HNH H 'N4 __INH

NH

2 0- 1

NH

2 HN IAN HN "H IN 34 H (0H 2 )2CH3 o 0 0 r_ 40 R =(CH 2 )pCHS 36H=(CH~CN 0-HYH~, 429=CH),CH3 AO/ H H HO H Boc-fR(NO,)-OMe(26) Boc-f-OII (7.64 g, 28.8 inmol), H-W(N0 2 )-OMe'HCI (6.72 g, 28.8 mnmol), HOBt (3.94 g, 29.2 mmol), TEA (8 ml, 57.6 mmol) and DMF (1 10 ml, anhydrous) are combined and cooled at 0 0

C

and EDCI (5.89 g, 30.8 mamol) is added with stirring. After stirring at room temperature for 18 hi, WO 02/26774 PCT/US01/30051 the mixture is concentrated by rotary evaporation, diluted with water (350 ml), and extracted with EtOAc (4x80 ml). The combined organic extract is washed with aqueous 1 N HC1 (3x60 ml), saturated aqueous NaHCO 3 (2x50 ml), brine (45 ml), and is then dried with anhydrous Na 2

SO

4 After filtration, the filtrate is concentrated by rotary evaporation, co-evaporated with ether ml), and dried in vacuo to give 3.

p-TSA H-fR(NO)-OMe (27) TFA (6 ml) is added to a solution of Boc-fR(N0 2 )-OMe (26) (1.49 g, 3.1 mmol) in DCM (20 ml) at 0°C. After stirring at 0°C for 30 min., the reaction mixture is stirred at room temperature for h. The solvent is removed in vacuo, the residue co-evaporated with ether (25 ml), and the resulting residue is dissolved in methanol (25 ml). p-toluenesulfonic acid monohydrate (0.6 g, 3.1 mmol) is added, the mixture stirred for 5 min. at room temperature, the volatiles removed in vacuo, and the residue co-evaporated with ether (2x25 ml) to give 27.

Boc-YfR(NO0)-OMe (28) p-TSA H-fR(N0 2 )-OMe (27, 3.46 g, 6 mmol), Boc-Y-OH (1.72 g, 6.12 mmol), HOBt (0.83 g, 6.18 mmol), TEA (1.6 ml, 12 mmol) and DMF (25 ml, anhydrous) are combined and cooled at 0 C and EDCI (1.1 g, 6.3 mmol) is added with stirring. After stirring at room temperature for 16 h, the mixture is concentrated by rotary evaporation, diluted with water (110 ml), and extracted with EtOAc (4x23 ml). The combined organic extract is washed with aqueous 1 N HC1 (3x20 ml), saturated aqueous NaHCO 3 (2x25 ml), brine (25 ml), and is then dried with anhydrous NazSO 4 After filtration, the filtrate is concentrated by rotary evaporation to give Boc-YfR(NO,)-OH (29) A mixture of Boc-YfR(N0 2

)-OCH

3 (28, 2.64 g, 4.1 mmol), LiOH (0.113 g, 4.72 mmol), water (0.4 ml) and MeOH (10 ml) is stirred at room temperature for 6 h. After removal of the solvent, the residue is dissolved in a minimum amount of water. 1 N HC1 (ca. 4.7 ml) is added to neutralize the mixture to pH 5-6. Filtration of the solid and drying in vacuo gives 29.

Boc-YfR(NO)W-NH, Boc-YfR(NO2)-OH (29, 0.99 g, 1.56 mmol), H-W-NH2-HCl (0.43 g, 1.8 mmol), HOBt (0.25 g, 1.84 mmol), TEA (0.55 ml, 3.9 mmol) and DMF (20 ml, anhydrous) are combined and cooled at 0°C and EDCI (0.36 g, 1.87 mmol) is added with stirring. After stirring at room temperature for 16 h, the mixture is concentrated by rotary evaporation, diluted with water (120 ml), and extracted with EtOAc (4x25 ml). The combined organic extract is washed with aqueous 1 N HC1 WO 02/26774 PCT/US01/30051 (3x20 ml), saturated aqueous NaHCO 3 (2x25 ml), brine (20 ml), and is then dried with anhydrous Na 2

SO

4 After filtration, the filtrate is concentrated by rotary evaporation, co-evaporated with ether (20 ml), and dried in vacuo to give H-YfR(NO,)-Trp-N (31) TFA (2 ml) is added to a solution of peptide 30 (0.95 g, 1.16 mmol) in DCM (6 ml) at 0° C. After stirring at 0° C for 30 min, the reaction mixture is stirred at room temperature for 5 h. The solvent is removed by rotary evaporation and the resulting residue is purified by preparative HPLC to give 31.

H-YfRW-NHI(32) Peptide 31 (0.25 g, 0.35 mmol), 5% Pd/BaSO 4 (0.25 g, unreduced) and MeOH (15 ml) are combined and hydrogenated under 40 psi of hydrogen at room temperature for 48 h. After filtration, the filtrate is concentrated by rotary evaporation and the residue purified by preparative HPLC to yield H-YfRW-NH2 (32).

Bc-YfR(NO,)W-NH (33) The procedure of preparing 28 is used, followed by purification by preparative HPLC and 33 is obtained from H-YfR(NO2)W-NH2 (31, 0.26 g, 0.37 mmol), butyric acid (0.037 g, 0.42 mmol), HOBt (0.059 g, 0.43 mmol), TEA (0.1 ml, 0.74 mmol), EDCI (0.084 g, 0.44 mmol), and DMF (12 ml).

Bc-YfRW-NH, (34) The procedure of preparing 32 is followed and 34 is obtained from 33 (0.16 g, 0.2 mmol), Pd/BaSO 4 (0.15 g, unreduced), MeOH (12 ml), and TFA (0.1 ml).

CH

3 (CH)CO-YfR(NO,)W-NH, The procedure of preparing 28 is used followed by purification by preparative HPLC and 35 is obtained from H-YfR(N0 2

)W-NH

2 (31, 0.26 g, 0.36 mmol), decanoic acid (0.071 g, 0.41 mmol), HOBt (0.057 g, 0.42 mmol), TEA (0.1 ml, 0.73 mmol), EDCI (0.083 g, 0.43 mmol), and DMF (12 ml).

CIICHL)CO-YfRW-NH, f36) The procedure of preparing 31 is followed and 36 is obtained from 35 (0.2 g, 0.23 mmol), Pd/BaSO 4 (0.18 g, unreduced), MeOH (12 ml), and TFA (0.1 ml).

Boc-YfR(N.)-tryptamide (37) WO 02/26774 WO 0226774PCT/USOI/30051 The procedure of preparing 30 is used and 37 is obtained from Boc-YfR(N0 2 )-OH (29, 0.95 g, rnmol), tryptamnine (0.276 g, 1.73 mmol), HOBt (0.239 g, 1.77 mmol), TEA (0.5 ml, 3.6 nimol), EDCI (0.34 g, 1.8 mmnol), and DMF (18 ml).

H-YfR(N0q)-tryptamide (38) Peptide 30 (0.98 g, 1.27 nmnol) in DCM (6 nil) and a solution of anisole-TFA-DCM 6 ni) are mixed at OTC and stirred with ice-cooling for 20 min. The mixture is then allowed to stir at room temperature overnight. The solvent is removed and the resulting residue is co-evaporated with ether (2x25 ml) to give crude 38.

Bc-YfR(N0 0 )-tryptamide (39) The procedure of preparing 28 is used followed by purification by preparative HPLC and 39 is obtained from 38 (0.28 g, 0.41 nimol), butyric acid (0.042 g, 0.47 mmol), HOBt (0.066 g, 0.49 iiol), TEA 12 nil, 0.83 mniol), EDCI (0.094g, 0.49 unnol.), and DMF (10 nil).

Bc-YfR(NO?)-trvptaniide The procedure of preparing 32 is followed and 40 is obtained from 39 (0.19 g, 0.25 rnol), PdIB aSO 4 18 g, unreduced), MeOll (12 ml), and TEA 1 ml).

C 3 (CHq) 8 CO-YfR(NO?,)-trvp2tamide (41) The procedure of preparing 28 is used followed by purification by preparative IIPLC and 41 is obtained from 38 (0.27 g, 0.4 mmol), decanoic acid (0.081 g, 0.47 mmrol), HOBt (0.065 g, 0.48 nimol), TEA (0.11 ml, 0.81 mmol), EDCI (0.093g, 0.49 mmol), and DMF (12 mlA).

CH

3 (CH,))CO-YfRW-trvptamide (42) The procedure of preparing 32 is followed and 42 is obtained from 41 (0.21 g, 0.25 mmol), 5 To Pd/B aSO 4 18 g, unreduced), MeOH (12 ml), and TEA 1 ml).

Examples 76 through 81 Syntheses of Ac-YfRW-OMe Ac-YfRW-NIICH 3 AcYfRWSar-NH 2 3-(4-OH- Ph)propanoy-fRW-NHCH 3 3-(4-OH-Phi)propanoyl-fRW-NH(CH 2 zH and 3-(4- O11-Ph)propanoyl-fRW-NIL(CJ~z) 2 0H These examples are prepared according to the plan presented in Scheme V: WO 02/26774 PCT/USO1/30051

UIOH

TFA

A -Y -O H

I

EDGI, HO~t TEA, DMF NH2 HN H4 N H A dPP HOJ 46

H

H2 LiOH 1) HNRIR2 IEDOI, HOBt TEA, DMF 2) H 2 53 Rl R2= OH 3 54 Rl H, R 2 CH CH2PH Rl CHI3 R2= CH~qONH 2 49 RI R2= OH3 Rl OHS R 2

=CHCONH

2 Scheme V Boc-fR(NOg,)-OHI (43) WO 02/26774 PCT/US01/30051 The procedure of preparing 29 is followed and 43 is obtained from Boc-fR(NO2)-OMe (26, 2.8 g, 5.8 mmol), LiOH (0.27 g, 11 mmol) and MeOH (15 ml).

Boc-fR(N0 9 )W-OMe (44) The procedure of 26 is followed and of 44 is obtained from Boc-fR(N0 2 )-OH (43, 4.22 g, 9.06 mmol), H-W-OMe HCI (2.65 g, 10.4 mmol), HOBt (1.44 g, 10.7 mmol), TEA (3.15 ml, 22.6 mmol), EDCI (2.08 g, 10.9 mmol), and DMF (55 ml).

H-fR(NO,)W-OMe TFA (12 ml) is added to a solution of 44 (5 g, 7.5 mmol) in DCM (30 ml) at 0°C. After stirring at 0°C for 1 h, the reaction mixture is stirred at room temperature for 16 h. The solvent is removed in vacuo, the residue co-evaporated with ether (2 x 35 ml), and the product dried in vacuo to give crude Ac-YfR(NO 2 )W-OMe (46) The procedure of preparing 28 is followed and crude 46 is collected from H-fR(N0 2 )W-OMe 3.3 g, 5.8 mnnol), Ac-Y-OH (1.5 g, 6.7 mmol), HOBt (0.93 g, 6.9 mmol), TEA (1.7 ml, 12.2 mmol), EDCI (1.34 g, 7 mmol), and DMF (25 ml).

Ac-YfRW-OMe (47) The procedure of preparing 32 is followed and 47 is obtained from 46 (0.2 g, 0.26 mmol), Pd/BaSO 4 (0.19 g, unreduced), MeOH (15 ml), and TFA (0.1 ml).

Ac-YfR(NO,)W-OH (48) The procedure of preparing 29 is used followed by purification by preparative HPLC and 48 is obtained from Ac-YfR(N0 2 )W-OMe (46, 0.96 g, 1.25 mmol), LiOH (0.063 g, 2.6 mmol), water (0.4 ml), and MeOH (6 ml).

Ac-YfRW-NHCH, (49) The procedure of preparing 30 is followed, exqept for the co-evaporation step, and a coupling product is obtained from Ac-YfR(NOz)W-OH (48, 0.3 g, 0.39 mmol), methyl amine (0.014 g, 0.45 mmol), HOBt (0.058 g, 0.43 mmol), TEA (0.092 ml, 0.91 mmol), EDCI (0.089 g, 0.47 mmol), and DMF(12 ml). This product is purified by preparative HPLC and is then hydrogenated using the procedure of preparing 32 with 5% Pd/BaSO 4 (0.17 g, unreduced) in MeOH (12 ml) to give 49.

Ac-YfRWSar-NH, WO 02/26774 PCT/US01/30051 The procedure of preparing 30 is followed, except for the co-evaporation step, and a coupling product is obtained from Ac-YfR(N0 2 )W-OH (19, 0.3 g, 0.39 mnmol), sarcosine amide'HC1 (0.05 g, 0.4 mmol), HOBt (0.058 g, 0.43 mmol), TEA (0.092 ml, 0.91 mmol), EDCI (0.089 g, 0.47 mmol), and DMF (12 ml). The product is purified by preparative HPLC and is then hydrogenated using the procedure of preparing 32 with 5% Pd/BaSO 4 (0.18 g, unreduced) in MeOH (12 ml) to give 3-(4-OH-Ph)propanovl-fRW-OCH, (51) The procedure of preparing 26 is used followed by purification by preparative HPLC to give 51 from H-fR(N0 2

)W-OCH

3 45 (2.1 g, 3.7 mmol), 3-(4-hydroxyphenyl)propanoic acid (0.71 g, 4.3 mmol), HOBt (0.59 g, 4.4 minol), TEA (1.1 ml, 0.91 mmol), EDCI (0.85 g, 4.5 mmol), and DMF ml).

3-(4-OH-Ph)propanovl-fRW-OII (52) The procedure of preparing compound 29 is followed to give 52 from 51 (1.15 g, 1.61 mmol), LiOH (0.081 g, 3.38 mmol), water (0.4 ml) and MeOH (8 ml).

3-(4-OH-Ph)propanovy-fRW-NHCH_ (53) The procedure of preparing compound 30 is followed to give a coupling product from 52 (0.3 g, 0.42 mmol), methylamine (0.015 g, 0.49 mmol), HOBt (0.064 g, 0.47 mmol), TEA (0.14 ml, 0.98 mmol), EDCI (0.098 g, 0.51 mmol), and DMF (12 ml). This product is purified by preparative HPLC and hydrogenated using the procedure of 32 with 5% Pd/BaSO 4 (0.16 g) and MeOH (12 ml) to give 53.

3-(4-OH-Ph)propanoyl-fRW-NHCH_9CHO0H (54) The procedure of preparing compound 30 is followed to give a coupling product from 52 (0.3 g, 0.42 mmol), ethanolamine (0.031 g, 0.51 mmol), HOBt (0.066 g, 0.49 mmol), TEA (0.14 ml, 1 mmol), EDCI (0.1 g, 0.52 mmol), and DMF (12 ml). This product is purified by preparative HPLC and hydrogenated using the procedure of preparing compound 32 with 5% Pd/BaSO4 (0.18 g) and MeOH (12 ml) to give 54.

3-(4-OH-Ph)propanoyl-fRWSar-NH The procedure of preparing compound 30 is followed to give a coupling product from 52 (0.3 g, 0.42 mmol), sarcosine amideHCl (0.061 g, 0.49 mmol), HOBt (0.068 g, 0.5 mmol), TEA (0.15 ml, 1.1 mmol), EDCI (0.098 g, 0.51 mmol), and DMF (12 ml). This product is purified by WO 02/26774 PCT/USOI/30051 preparative HPLC and hydrogenated using the procedure of preparing compound 32 with Pd/BaSO 4 (0-18 g) and MeOH (12 nil) to give.

Examples 82 through 84 Syntheses of Bc-YfRW-NHCH 3 Bc-YfRW-NH(CH 2 2 011 and BcYfRW-N(CH 3

)(CH

2 2 0H These examples are prepared according to the plan presented in Scheme VI:

NNO

2 -L NO 2 NtNN0 0

NH

2

NH

OH H-WOM H H N TFA RN 0HN 0 TEA,

DMF

HH N

HH

29 2057 HiH HO N- N02 NNO 2 Y ,0 X H2 0 NH 2

NH

2 Butyic Acid ,~HMSR R "DOI, HOBt LiOH KEDOI, ON TEA, OMF HN HN HN HRTA, OMF RN HN H N N N! 5J H 58 HO 59HO H ~60 Rl=H, R 2 =CH3 N NH 61l R 1

R

2

OHOH

2

OH

12 ~62R P 1 3 2

R

2 -CH-12CH

H

2 Pd-BaSO 4

?H

0

FI

0 H 6 R1 R2= GH3HG HO

CH

2

CH

2

OH

Scheme V1 Boc-YfRW-OCH 3 (56) The procedure of preparing compound 28 is followed and 2.1g of 56 is obtained from Boc- YfR(N0 2 )-Oll (29, 1.7 g, 2.6 mmol), H-W-OCH 3 *HCI (0.68 g, 2.7 mmol), HOBt (0.42 g, 3.1 mmol), TEA (0.8 ml, 5.8 mmol), EDCI (0.63 g, 3.3 mmol), and DM4F(25 ml).

II-YfRW-OCH 3 (57) The procedure of preparing compound 38 is followed and 57 is obtained from 56 (2.1 g, mmol) and a solution of anisole-TFA-DCM 18 ml).

B3c-YfRW-OCH 3 (58) 'The procedure of preparing compound 28 is followed to give a crude coupling product from H- YfR(N0 2

)W-OCH

3 (57,1.21 g, 1.65 nimol), butyric acid (0.18 g, 2 nimol), HOBt (0.26 g, 1.95 WO 02/26774 PCT/US01/30051 mmol), TEA (0.57 ml, 4.1 mmol), EDCI (0.394 g, 2.06 mmol), and DMF (25 ml). This product is purified by preparative HPLC to give 58.

Bc-YfRW-OH (59) The procedure of preparing compound 29 is followed to give 59 from 58 (0.1 g, 0.13 mmol), LiOH (0.004 g, 0.15 mmol), water (0.2 ml) and MeOH (2 ml).

Bc-YfRW- NHCH 3 (63) The procedure of preparing compound 28 is followed to give a coupling product 60 from 59 (0.1 g, 0.125 mmol), methyl amine (0.006 ml, 0.15 mmol), HOBt (0.02 g, 0.15 mmol), TEA (0.04 ml, 0.3 mmol), EDCI (0.03g, 0.16 mmol), and DMF (3 ml). This product is hydrogenated according to the procedure of preparing compound 32 with 5% Pd/BaSO 4 (0.1 g, unreduced) in MeOH (6 ml) to give 63.

Bc-YfR(NO,)W- NH(CH 2 OH (61) The procedure of preparing compound 28 is used followed by purification by HPLC to give 61 from 59 (1.3 g, 1.65 mmol), ethanolamine (0.12 ml, 1.9 mmol), HOBt (0.263 g, 1.95 mmol), TEA (0.55 ml, 4 mmol), EDCI (Q.38g, 2 mmol), and DMF Bc-YfRW- NH(CH)70OH (64) The procedure of preparing compound 32 is followed and 64 is obtained as a solid from 61 (0.22 g, 0.26 mmol), 5% Pd/BaSO 4 (0.2 g, unreduced), MeOH (12 ml), and TFA (0.1 ml).

Bc-YfR(NO)W-N(CH )(CH )OH (62) The procedure of preparing compound 28 is used followed by purification by HPLC to give 62 from 59 (0.26 g, 0.33 mmol), N-methylethanolamine (0.032 ml, 0.4 mmol), HOBt (0.053 g, 0.39 mmol), TEA (0.11 ml, 0.83 mmol), EDCI (0.076g, 0.4 mmol), and DMF (10 ml).

Bc-YfRW-N(CH 3 )(CH )OH The procedure of preparing compound 32 is followed and 65 is obtained from 62 (0.16 g, 0.19 mmol), 5% Pd/BaS04 (0.19 g, unreduced), MeOH (10 ml), and TFA (0.1 ml).

Example Synthesis of 3-(4-OHPh)propanoyl-YfRW-NH2 (69) This example is prepared according to the plan presented in Scheme VII: WO 02/26774 WO 0226774PCT/USOI/30051 H HH NH2 N 2

N..N

N2 propanoic acid -0 If NH 0 N-12' EaD, HOBt IIO Me TEA,. ONMe L FIN 0 H TEA,1- DM H 0HN 0 66 67 HO HP H H NH2NNH

NH

2 NH 0. N V-'k 0 ECCI HO~t MN 4 HN 2 1 Pd-BaS0 4 H4 R TEA, DMVF 0 N0H 8 N' \9 N H H J

H

Scheme VII 3-(4-OB~h)propanoy-fR(NO)-OCH 66 The procedure of preparing compound 28 is used followed by purification by preparative IAPLC to give 66 from 27 (1.5 g, 2.6 mmol) 3-(4-hydroxyphenyl)propanioic acid (0.48 g, 2.9 mmol), IlOBt (0.43 g, 3.2 mmol), TEA (0.91 n-l, 6.6 mmol), EDGI (0.63 g, 3.3 minol), and DMF (28 ml).

3-(4-Q1HPh)propanoyl-fR(N0 2 )-OH (67) The procedure of preparing compound 29 is followed to give 67 from 66 (0.33 g, 0.63 nimol), LiOH (0.029 g, 1.2 mmol), water (0.4 mlA) and MeOH (5 mlA).

3-(4-OBPh)prop~anoyl-fR(NO0 2 )W-NII, (68) The procedure of preparing compound 28 is used followed by purification by preparative HPLC to give 68 from 67 (0.32 g, 0.62 mmol), H-W-NH2*HCl (0.16 g, 0.68 nimol), HOBt (0.1 g, 0.73 nimol), TEA (0.2 ml, 1.6 mmol), EDCI 14g, 0.75 nimol), and DMF (13 ml).

3-(4-OH~h)propanoyl-fRW-NH 2 (69) The procedure of preparing compound 32 is followed and 69 is obtained from 68 (0.14 g, 0.2 Inmol), 5% Pd/33aSO 4 (0.12 g, unreduced), MeOH (9 ml), and TFA 1 nil).

Example 86 Synthesis of Ae-YfR-tryptainide (73) This example is prepared according to the plan presented in Scheme VIII: WO 02/26774 PCT/USOI/30051

H

NH0 W27 Ac-Y-OH EDGI. HO~t TEA, DMF

H

Tryptamnile 0 HO~t HN aHN TEA, DME H NHAc N

HO

H H 6> I>N N02 N NN0

NH

2 H HH 0 HAc NHAC 70 7/1~

HO

oHN N N A3

H

HO

Scleme Vill Ac-YfR(N09)-0CH-, The procedure of preparing compound 28 is used followed by purification by preparative I-ILC to give 70 from 27 (0.36 g, 0.63 mmol), Ac-Y-O11 14 g, 0.63 mmcl), HOBt 1 g, 0.74 mmol), TEA (0.17 nil, 1.2 mmol), EDCI (0.14 g, 0.76 mmol), and DMF (12 mlA).

Ac-YfP(N02)-OH (71) The procedure of preparing compound 29 is followed with an additional co-evaporation step from THE (5 ml) to give a mixture of 71 and two equivalents of LiCi from 70 (0.08 g, 0.14 mmol), LiOH (0.007 g, 0.28 mmol), water (0.2 ml), and MeOH (4 ml).

Ac-YfR-tryptamide (73) The procedure of preparing compound 28 is followed to give a crude coupling product 72 from the mixture of Ac-YfR(N0 2 )-OW2LiCI (71, 0.091 g, 0.136 mmol), tryptamnine (0.025 nil, 0.15 mmol), HOBt (0.021 g, 0.15 rmnol), TEA (0.1 ml, 0.7 mmol), EDCI (0.03g, 0.16 nimol), and DMLF (6 ml). This product is hydrogenated using the procedure of preparing compound 32 using 5% PdIBaSO 4 (0.2 g, unreduced) and MeOH (8 ml) to give 73 as a solid.

Example 87 Synthesis of Hydrocinnamoyl-fRW-NU2 (77) This example is prepared according to the plan presented in Scheme IX: WO 02/26774 PCT/USOI/30051

H

N N

OPAO

ITSP

W

2 7 0 Hydrocinnamic Acid EDCI, HOBt TEA, DMF 0rJ NH 2 HN H 0 0 74 UGH .(LN yOHNH HN 0H

H

N N N0 N2

H-W-NH

2

O

SEDC, HO~t HN aHN TEA, DMVF IN~- 76

N

H

H

N NH

H

2 /Pd-BRS0 4

NH

77

H

Scheme IX Hydr ocinnamoyl-fR(NOj -OCH 3 (7-4) The procedure of preparing compound 28 is followed to give 74 from 27 (0.25 g, 0.43 mmol), hydrocinnamic acid (0.068 g, 0.45 nmol), HOBt (0.068 g, 0.5 mmrol), TEA (0.16 ml, 1.1 rumol), EDCI (0.097 g, 0.51 nnnol), and DNM (20 m) Hydrocinnamoyl-fW(NO,)-OH The procedure of preparing compound 29 is followed to give 75 from 74 (0.22 g, 0.43 mmol), LiOl (0.014 g, 0.56 mmol), water (0.3 ml) and MeOH (8 nml).

Hydrocinnamnoyl-fRW-NH 9 (77) The procedure of preparing compound 28 is followed to give a crude coupling product 76 from (0.2 g, 0.4 nunol), H-W-NH2'HCI 1 g, 0.4 2 numol), HIOB t (0.068 g, 0. 5 Mmnol), TEA 14 nml, 1 nimol), EDCI (0.1 g, 0.52 rnmol), and DMF (12 ml). This product is hydrogenated using the procedure of preparing compound 32 using 5% Pd/BaSO 4 (0.2 g, unreduced) and MeOH nil) to give 77.

Examiple 88 Synthesis of 3-(4-OHPh)propanoyl-(4-F-f)RW-NH 2 (82) This example is prepared according to the plan presented in Scheme X: WO 02/26774 WO 0226774PCT/USOI/30051

F

H F HF N N NO2 'r NN2 INN 0 0o NH 2 NH f1 NN2 I HO~ 0HO TAe H Hi OMe TEA4 DMF I. e tGMG BoCNH H0 0 78 79 F H F H F NH2 N2 N H 3~ OHhdoyhey) 001 H-W-NH 2 0 H EDpaoi ci d UOH ON FOCi, HoBI 'rt' 0 I.J O HNO r H TEA DMF H YEA, DMF a\ 'OH 2) H 2 Pd-BaSO4 a HN NH'

N

H

80 81 82 HO HO H Sohemox B oc- (4-F-f)R(NO,)-OMe (78) The procedure of preparing compound 26 is followed to give 78 from Boc-(4-F-f)-OH (1.5 g, 5.3 inmol), HR(NO 2 )-OMte-HCl (1.46 g, 5.4 imniol), HOBt (0.87 g, 6.4 imcd), TEA (1.86 mul, 13.3 mmol), EDCI (1.33 g, 6.9 mmol), and DMF (35 ml).

H-(4-F-f)R(NO2)-OMe (79) TFA (4 a) is added to a solution of 78 (2.48 g, 5 mmol) in DCM (20 ml) at 0 0 C. The mixture is stirred with ice-cooling for 30 min. and then is stirred at room temperature overnight. The solvent is removed, the residue co-evaporated with ether (2030 ml), and the resulting residue is dried under vacuum to give 79 as a solid.

3-(4-OHPh)propanoyl-(4-F-f)R(N0 9 )-OMe The procedure of preparing compound 26 is followed to give 80 from 79 (2 g, 5 mmol), 3-(4hydroxyphenyl)-propanoic acid (0.92 g, 5.5 mmol), HOBt (0.8 g, 5.9 mmol), TEA (1.7 ml, 12.6 mmol), EDCI 15 g, 6 mmol), and DMF (25 mld).

3-(4-OH~h)propanov1-(4-F-f)R(NO,))-OH (81) The procedure of preparing compound 29 is followed to give 81 from 80 (0.75 g, 1.4 nunol), LiOH (0.065 g, 2.7 mmoL), water (0.4 ml), and MeOH (8 mlA).

3-(4-O1{Ph)propangyl-(4-F-flR(NO,)W-NH (82) The procedure of preparing compound 30 is used followed by purification by preparative 1{PLC to give a coupling product from 81 (0.4 g, 0.75 mmol), H-W-NH2HCl (0.2 g, 0.82 mmol), HOBt (0.12 g, 0.9 mmol), TEA (0.26 ml, 1-9 nunol), EDCI (0.17 g, 0.9 mmol), and DMF (20 This product is hydrogenated using the procedure of preparing compound 32 using 5% Pd/B aSO 4 (0.25 g, unreduced) and MeOH (15 ml) to give 82.

WO 02/26774 PCT/USOI/30051 Examples 89-91 Syntheses of 3-(4-OHPh)propanoy-TR-tryptaniide 3-(2-OHPh)propanoyl-flRtryptanide and Hydrocinnamoyl-f.R-tryptamide These examples are prepared according to the plan presented in Scheme XI: HH H _NN N N N N _fN0 2 Y -NO2 Z2-~NO 2 LK~J NH 2 NH2a 'f N2 tryptamine NH EH CI, HOST 0 1) Boc NH H0 TEA, DMF~ BOCNH H4 2) p-TSA-H N N3H0 43 8i HN_- 84 H 'N N HH H

H

4 N Y N N N r NH 0 '4j 0 ArOOH H H2/P-BS H4 EDC1, HOBT aN HN 0H NMM, DME F N Y N ey H ey H X 85 X=OH,Y=H X 88 X=OH,Y=H BSX=H,Y=OH 89 X=HY=OH 87X=Y=H Scheme XI BocfR(N0 7 )-tryptamide, (83) Boc-fR(N0 2 )-OH (43, 507.9 mg, 1.09 mmol), tryptamnine (172.9 mg, 1.08 nimol), HOBt (163.3 g, 1.08 mmol), TEA 16 ml, 0. 12 nimol) and DMEF (6 ml, anhiydrous) are combined and cooled at OTC and EDGI (226.9 mg, 1. 18 mmol) is added with stirring. After stirring at OTC for 45 min the ic0 ice bath is removed and the mixture is warmed and stirred at room temperature for 14.5 h. The mixture is diluted with EtOAc (20 mlA) and washed with aqueous 2 N HCl (3x5 ml), the aqueous acid layer is back-extracted with EtOAc (lx 10 ml), and the combined EtOAc layers are washed with I M NaHCO 3 (3x5 ml) and brine (10 ml). The organic extract is then dried with anhydrous Na 2 S 04, the dessicant is removed by filtration, and the filtrate is concentrated by rotary. This solid is dried in vacuo to give 83.

p-TSA e H-fR(NO2)-trytamide (84) TFA (3 mnl) is added to a solution of Boc-IR(N0 2 )-tryptamide (83) (0.58 g, 0.95 nimol) in DCM (6 ml) at 0 0 C. After stirring at OTC for 30 min., the reaction mixture is stirred at room temperature for 1.25 h. p-Toluenesulfonic acid monohydrate (176.2 mug, 0.93 mmol) is added and the volatiles are removed by rotary evaporation to leave a brown oil. The oil is triturated with ether (10 ml) and the residue dried in vacua to 84.

3-(4-OH~h)propanovl-fR(N&,')-tryvtamide WO 02/26774 WO 0226774PCT/USOI/30051 p-TSA H-fR(N0 2 )-tryptamide(84, 149.9 mg, 220 Lrol), 3-(4-hydroxyphenyl)propanoic acid (39.6 mg, 238 limiol), HOBt (33.7 mg, 233 gino1), NMM (0.37 nil, 337 jtmol) and DMEF (1.5 nil, anhydrous) are combined and cooled at 0 0 C and EDCI (44.6 mg, 233 tmol) is added with stirring. After stirring at 0 0 C for 1.3 h the ice bath is removed and the mixture is warmed and stirred at room temperature for 100 h. The muixture is diluted with FtOAc (20 nil) and washed with aqueous 2 N HCl (3x5 ml), the aqueous acid layer is back-extracted with EtOAc (lx 10 all), and the combined EtOAc layers are washed with 1 M NaHICO 3 (3x5 nil) and brine (10 ml). The organic extract is then dried with anhydrous Na 2 S 04, the dessicant is removed by filtration, and the filtrate is concentrated by rotary evaporation to give 3-(2-01IPh)propanoyl-fR(NO,)-trvpta-ilde (86) The procedure used in preparing compound 85 is followed to give crude coupling product from 84 (151.7 mg, 223 .rmmo1), 3-(2-hydroxyphenyl)propanoic acid (39.2 mg, 236 tmnol), HOBt (35.3 mg, 231 iol), NMAM (0.40 ml, 364 jimol), EDCI (45.0 mg, 235 A.mol), and DMF (1.5 ml, anhydrous). This material is further purified by preparative 1LPLC (C 4 to afford 86.

I-1ydrocinnamoyl-fR(NO 9 2)-tryptamide (87) The procedure used in preparing compound 85 is followed to give 87 from 84 (150.6 mg, 221 ginmmol), hydrocinnamic acid (35.6 mag, 237 tmol), ROMt (34.4 mig, 225 jimol), NMM (0.37 ml, 337 j tmol), EDCI (45.1 mg, 235 uniol), and DMF (1.5 nml, anhydrous).

3-(4-OH~h)propanovl-fR-trvtamide (88) The procedure used in preparing compound 32 is followed to give 88 from 95 mg (145 iol) of and 5% Pd-BaSO 4 (68 mg, unreduced).

3-(2-OHF'h)nrop~anoyl-fR-tryntatrnide (89) The procedure used in preparing compound 32 is followed except that the product is lyophilized from acetonitrile-water rather than purified by HPLC to give 89 from 33.7 mig (5 ljimol) of 86 and 5% Pd-B aSO 4 (31 mng, unreduced).

llvdrocinnamoyl-fR-trytamide The procedure used in preparing compound 32 is followed to give 90 from 106 mg (165 Rmol) of 87 and 5% Pd-BaSO 4 (63 mg, unreduced).

Example 92 Synthesis of 3-(4-OHPh)propanoyl-Me-fR..tryptanmide WO 02/26774 PCT/USOI/30051 This example is prepared according to the plan presented in Scheme XII: H H H Ni N N0 2 N <N NN N NN02 OH+ N2EDOIHO~t 0 H a1)TFA.

H

2 N OMe TEA, DMF .2NA.Oe 2) p-TSAI-H BC0Boe N, H o 91 p-TSA 92 H H H N N~H N N, N NH 3.4.t~ohn~'Z"O NH NH2 NH p-4roxp 1O eO tryptamine B- N EDCI, HO~t EDCI, HO t -H 0 LOH _ijNMM, DMF NMM, DMF 0 0 2)H 2 /Pd-BaGO 4

H

?94 95 H H0 HO HO~r Schame XII Boc-Me-fR NO2>-OMe (91) Boc-Me-f-OH (1.0008 g, 3.6 mmol), H-R(N0 2 )-OMe *HCI (0.9659 g, 3.6 nunol), HO~t (0.5515 6 g, 3.6 mmol), TEA (0.525 ml, 3.8 mmol) and DMF (20 ml, anhydrous) are combined and cooled at OTC and EDCI (0.7211 g, 3.8 mmol) is added with stirring. After stirring at OTC for 1 h the ice bath is removed and the mixture is warmed and stirred at room temperature for 23 h. The mixture is diluted with EtOAc (20 ml) and washed with aqueous 2 N HCl (3x8 ml), the aqueous acid layer is back-extracted with EtOAc (Ix 10 ml), and the combined EtOAc layers are washed with 1 M NaHCO 3 (3x8 ml) and brine (10 ml). The organic extract is then dried with anhydrous Na 2

SO

4 the dessicant is removed by filtration, and the filtrate is concentrated by rotary evaporation to give 91.

p-TSA H-Me-fR(NG+)-OMe (92) The procedure of making compound 84 is followed to give 92 as a hydrated p-TSA salt from 91 (0.4989 g, 1.0 mmol).

3-(4-OBPh)propanoyl-Me -fR(NO2)-OMe (93) p-TSA H-Me-fR(N0 2 )-OMe (92, 282.8 mg, 500 p.tmmol), 3-(4-hydroxyphe-nyl)propanoic. acid (85.7 mg, 516 .tmol), HOBt (77.4 mg, 505 [tmol), NMM (0.60 ml, 546 jimol) and DMF (3 ml, anhydrous) are combined and cooled at 0 0 C and EDCI (101.9 mg, 532 jimol) is added with stirring. After stirring at OTC for 1.3 h the ice bath is removed and the mixture is warmed and stirred at room temperature for 24 h. More EDCI (41.2 mg, 215 jimol) is added, the mixture is stirred at room temperature for 64 h, EDCI 29.9 mg, 156 jimol) is again added along with more NM (0.60 mlA, 546 jimol), and the mixture is heated at 50'C for 8 h. The mixture is then left at WO 02/26774 PCT/US01/30051 room temperature for 21 d and is worked up as described in the procedure of preparing compound 91 to give 93.

3-(4-OHPh)propanoyl-Me-fR(NO,)-OH (94) A mixture of 3-(4-OHPh)propanoyl-Me-fR(NO2)-OMe (93, 96.3 mg, 177 Ltmol), LiOH monohydrate (21.3 mg, 508 pmol) in THF-MeOH-water (6 ml, 4:1:1) is stirred at room temperature for 1 h. 6% aqueous KHSO 4 (1.3 ml) is added and the volatiles are removed by rotary evaporation. Water (1.4 ml) is added to the wet residue, the pH adjusted to ~2 with a few more drops of 6% aqueous KHSO 4 and the aqueous mixture is extracted with EtOAc (3 x 2 ml).

The combined organic layers are washed with water and brine and are dried over anhydrous Na 2

SO

4 The spent dessicant is removed by filtration and the filtrate is concentrated by rotary evaporation to give 94.

3-(4-OHPh)propanoyl-Me-fR-tryptamide 3-(4-OHPh)propanoyl-Me-fR(NO 2 )-OH (94, 77.0 mg, 146 jtmol), tryptamine (23.9 mg, 149 Limol), HOBt (25.1 mg, 164 imrol), NMM (0.17 ml, 155 pmol) and DMF (5 ml, anhydrous) are combined and cooled at 0 C and EDCI (32.4 mg, 169 pmol) is added with stirring. After stirring at 0 C for 1.5 h the ice bath is removed and the mixture is warmed and stirred at room temperature for 4 h. The mixture is worked up as in the procedure for preparing compound 85 to give a crude coupling product which is purified by preparative HPLC(C 4 The purified product is then dissolved in 10% acetic acid-MeOH (20 ml) and is hydrogenated under 40 psi of H2 at room temperature for 15 h using 5% Pd-BaSO 4 (31.3 mg, unreduced) as catalyst. The catalyst is removed by filtration through Celite, the volatiles are removed in vacuo, the residue is redissolved in 10% acetonitrile-water (15 ml), and the mixture is frozen and lyophilized to yield Example 93 Synthesis of 5-[2-Acetylamino-3-(4-chloro-phenyl)-propionylamino]-4-oxo-6-phenyl-2-(2pyridin-2-yl-ethyl)-hexanoic acid [1-carbamoyl-2-(1H-indol-3-yl)ethyl]-amide (103) This example is prepared according to the plans presented in Schemes XII and XIV: WO 02/26774 WO 0226774PCT/USOI/30051 0 )CIOt TEA (DN C 0

'N

OH 2) CHNp

-N'

NHNH NH CI Nal a1 BNH 8c Boc'C 98 97 co 2 Me *N

H

2

(CO

2 Me) 2 'NO e BO N 9 0 M N.OM. CO 2 Me I N N 98 CO 2

MO

1) NaCH, MeCH 0 2) toluene, ref lux0 10OH Scheme XIII 3-(tert-Butoxycarbnylanino-1--diazo-4zphenylbutan-2-one (96) To a solution of Boc-f-OI-I (5.30 g, 20 nimol) in dry TI-IF (100 ml) are added TEA (2.02 g, nimol) and ethyl chloroforinate (2.16 g, 20 mmol) at -15'C under argon atmosphere. The mixture is stirred at C for 30 miii, and is then warmed to 0 0 C. A solution of diazomethane in ether [100 nml, prepared from Me(NO)NCONH2 (4.0 g, 40 nimol) and 50%7 aqueous KOH (20 nil)] is added. The mixture is warmed and stirred at room temperature for 3 h. The mixture is washed with saturated aqueous NaHCO 3 (30 nml), saturated aqueous amonium chloride (30 ml), and brine (2 x 30 mlA). The organic layer is dried over Na 2

SO

4 and concentrated by rotary evaporation. The residue is crystallized from hexanes-EtOAc at 5 0 C to give desired product. The mother liquor is concentrated and purified by chromatography (hexanes-EtOAc, 80;20) to give additional desired product.

(R)-3-(tert-Butoxycarbonyjamiino)-l-chloro-4-phenylbutan-2-one (97) To a solution of diazoketone (96, 115 mg, 0.4 mmol) in ether (3 mI) is added 4N~ HCII1 ,4-dioxane 11 ml, 0.44 nimol, prepared by diluting conc. HCl with 1,4-dioxane) dropwise at (f C. The resulting mixture is stirred at 0 0 C for 30 min. Several drops of TEA are added to neutralize the solution and the mixture is diluted with EtOAc (20 ml), washed with saturated aqueous NaHICO 3 mlA), and brine (3 x 10 ml). The organic layer is dried over Na 2

SO

4 and the solvents are evaporated. The residue is purified by chromratography (hexanes-EtOAc, 90: 10) to give 108 mg of a colorless solid: nip.: 101-102 'H NMR (CDC1 3 400 Mvllz) 6 1.43(s, 9H1), 3.03, 3.10 (AIIX, JAB 13.8 Hz, JAx 7.1 Hz, J-BX 6.8 Hz, 211), 4.00, 4.19 (AB, JAB 16.2 Hiz, 2H1), 4.70 (in, 114), 5.04 (br d, J =6.8 Hz, 111), 7.18-7.37 (in, 511).

WO 02/26774 PCT/US01/30051 Methyl 2-methoxvcarbonyl-4-(2'-pyridinyl)butanoate (98) To freshly prepared NaOMe [from 2.3 g (0.1 mol) of sodium] in MeOH (25 ml) is added dimethyl malonate (32 g, 0.24 mmnol). A solution of 2-vinylpyridine (10.5 g, 0.1 mol) in MeOH ml) is added dropwise over 40 min to the NaOMe solution at reflux. The resulting mixture is heated at reflux for 2.5 h. MeOH is removed under reduced pressure, the residue is treated with 2N HCI (150 ml), then is extracted with ether (2 x 60 ml) to remove excess dimethyl malonate.

The aqueous phase is made basic with 2N NaOH and extracted with ether (3 x 100 ml). The combined organic phases are washed with brine (3 x 60 ml) and dried over Na 2 S0 4 The filtrate is concentrated under reduced pressure, then most of the excess 2-vinylpyridine is removed in 1o vacuo. The residue is purified by chromatography (hexanes- EtOAc, 60:40 to 50:50 to 30:70) to give desired product.

Methyl (R)-5-(tert-Butoxycarbonylanino)-2-methoxvcarbonyl-6-phenyl-2-[2'-(2"pyridinvl)ethyll-4-oxohexanoate (99) To a solution of the c-chloroketone (97, 150 mg, 0.5 mmol) in dry 1,2-dimethoxyethane (3.0 ml) is added Nal (75 mg, 0.5 mmol) and the mixture is stirred under an argon atmosphere for 15 min (Mixture To a solution of the diester (98,142 mg, 0.6 mmol) in dry 1,2-dimethoxyethane ml) is added freshly prepared NaOMe (32 mg, 0.6 mmol) and the mixture is stirred under an argon atmosphere for 15 min (Mixture Mixture B is added to mixture A and the resulting mixture is stirred at room temperature for 1 h. The mixture is diluted with EtOAc (30 ml) and washed with brine (2 x 10 nil). The organic layer is dried over Na 2

SO

4 the solvents are removed by rotary evaporation, and the residue is purified by chromatography (hexanes-'PrOH, 80:20) to give 99.

(2RS, 5R)-5-(tert-Butoxycarbonvlamino)-6-phenvl-2-F2'-(2"-pyridinyl)ethyll-4-oxohexanoic acid (100) To a solution of di-ester (99, 165 mg) in MeOH (10 ml) is added 5N NaOH (1.0 ml) and the resulting mixture is stirred at room temperature for 2 h. The mixture is concentrated by rotary evaporation and the residue is dissolved in water (10 ml) and acidified with 3N HCI to pH 3.

The mixture is extracted with DCM (3 x 10 ml) and the combined organic phases are washed with brine (10 ml) and dried over Na 2

SO

4 The solvents are removed by rotary evaporation to give the di-acid. The crude di-acid is suspended in toluene (10 ml) and the mixture is heated at reflux under an argon atmosphere for 3 h. The solvent is evaporated and the residue is purified by WO 02/26774 WO 0226774PCT/USOI/30051 chromatography (DCM-MeOH, 98:2 to 95:5) to give a mixture of two diastereoisomers that are separable by preparative HPLC.

I- 'H 0 Ac-(4-C F)-OH

N

FDI, Hoat TFA 0 oarI HO~t TEA, OMF 0TEA, DMF0 B HN NH 2 MNN MN NN NHNHO 3N ieO 101 102 H S-heme XIV -B enzy-4-f 1-carbamoyl-2-(1H-indol-3-yl)-ethylcarbamoyll-2-oxo-6-pyridin-2-:yl-hexvllcarbamnic acid tert-butyl ester (101) Acid 100 (44 mg, 0.1 mmol), H-W-NI4jHCl (26 mg, 0.11 nimol), HOBt(16 mg, 0.12 mmol), TEA (24 mg, 0.24 mmol), EDCI (24 mg, 0.12 mmol) and DIVI(1 ml, anhydrous) are combined.

The mixture is stirred at room temperature overnight, then poured into water (10 ml) and extracted with EtOAc (4x9ml). The combined extract is washed with 1 N HC1 (2x8 ml), saturated NaHCO 3 (Ix 8 ml), brine (8 ml) then dried with anhydrous Na 2

SO

4 After filtration, the filtrate is concentrated by rotary evaporation to give 101.

5-A-mino-4-oxo-6-phentyl-2-(2-pyridiin-2-yl-etyhexanoic acid f I-carbamoyl-24( 1H-indol-3yl)ethvll-amide(102) Peptide 101 (97 mg, 0.16 mmol) is mixed with a solution (1 mlA) of anisole-TFA-DCM The mixture is stirred at room temperature overnight. After removal of solvent, the resulting residue is co-evaporated with ether (2xg ml) to give 0.81 g (99 of 102.

Ac-(4-Cl-F)-OH A mixture of Boc-(4-Cl-F)-OH (54 mg, 0.18 mmol), DCM (0.5 mld), and TFA (0.5 ml) is stirred at room temperature for 2 h. After removal of solvent, the resulting residue is mixed with DCM (1 acetic anhydride (17 mg, 0.2 mmol), and TEA (40 mg, 0.4 mmol). The mixture is stirred at room temperature overnight, then concentrated by rotary evaporation to yield Ac-(4-Cl-F)-OH.

5-[2-Acetyanmino-3-(4-chloro-phenyl)-:propionylaniinol-4-oxo-6phenl-2-(2pyridin-2:yl-ethl) hexanoic acid fl-carbamoyl-2-(IH-indol-3-l)ethyll-amide (103) EDCI (36 mg, 0.19 mmol) is added to a mixture of peptide 102 (81 mg, 16 mmol), Ac-(4-Cl-F)- OH (41 mg, 0.17 nimol), HOBt (24 mig, 0.18 mmol), TEA (35 mng, 0.35 mmol), and.DMI(2 ml, anhydrous) at 0 0 C. The reaction mixture is stirred at room temperature overnight and then is WO 02/26774 PCT/USOI/30051 worked up as in the procedure for making compound 101. The crude product is purified by preparative HPLC to give 103.

Example 94 Synthesis of N-{3-[9-Benzyl-12-(4-chlorobenzyl)-3-(1 H-Jndol-3ylmethyl-2,5,8,14-tetraoxo- 1,4,7,13-tetraaza-cyclotetracos-6-yl]-propyl}-guanidine (110) This example is prepared according to the plan presented in Scheme XV: NH NH NH PN NH PikNJNH PbN )NH H H H N HI N N Hp -A0 NN BO' MeG o 104 105 Doc

D

NH NH NH

P

4 NH N NH Pbf N NHJN TH 0H 0H N H Fmoo-li-Aun-OH NH DC, HO~t H 1) TFA H N o NHNMM, DMF NH 2) p-TSA-H NH C1M80 Nor MeD 1 MeG NH Emoc NH 106 Pbf, N A-NH HN NMH H N s I"~ 2; GDCI, HORt NH NH NH NMM, DMF CI HN C; Z H HN NH N Scheme XV 109 110 Boc-R(Pbf)W-OMe (104) NMM (0.48 ml, 4.4 mrmol) is added to a mixture of Boc-R(Pbf)-OH (1.053 g, 2.0 mmol), HA- OMe liCl (0.509 g, 2.0 unal), HOBt (0.270 g, 2.0 mmol), EDCI (0.422 g, 2.2 nimol) in DMF mlA) at 0 0 C and the resulting mixture is stirred at 0 0 C for 1 h and then at room temperature for 4 hi. The reaction mixture is diluted with EtOAc (100 ml) and is washed successively with water (2 x 10 ml), 1 N HCI (2 x 10 ml), saturated NaHCO 3 2 x 10 ml), and brine (2 x 10 The organic layer is dried over MgSO 4 the dessiuant removed by filtration, and [he filtrate concentrated by rotary evaporation to yield 1.454 g (100%) of crude 104.

n-TSA a H-R(Pbf)W-OMe (105) WO 02/26774 PCT/US01/30051 A solution of 104 (1.454, 2.0 mmol) in a mixture of TFA-DCM-water (10:40:0.5, 20 ml) and DCM (20 ml) is stirred at room temperature for 6 h. p-Toluenesulf-onic acid monohydrate (0.380 g, 2.0 mmol) is added and the mixture stirred for 10 min at room temperature. The solvent is removed by rotary evaporation and the residue is triturated with ether-hexane 100 ml) to yield 1.598 g of crude 105.

Boc-(carba-4-C1-Ff)-R(Pbf)W-OMe (106) EDCI (105.5 mg, 0.55 mmol) and NMM (0.12 ml, 1.1 mmol) are added to a well-stirred mixture of 5 (216 mg, 0.5 mmol), 105 (399.5 mg, 0.5 mmol), and HOBt (67.6 mg, 0.5 mmol) in DMF (9 ml) at o0C. The resulting mixture is stirred at 0°C for 1 h and then at room temperature for 5 h.

The mixture is worked-up as in the procedure for preparing compound 104 to give 508 mg (98%) of 106.

p-TSA H-(carba-4-C1-Ff)-R(Pbf)W-OMe (107) A solution of 106 (508 mg, 488 pmol) in TFA-DCM-water (10:90:0.5, 15 ml) is stirred at room temperature for 17 h. Analysis by HPLC indicates that the reaction is not yet complete so more TFA-DCM-water (10:90:0.5, 10 mi) and water (1 ml) are added and the resulting mixture is stirred at room temperature for 7 d. p-Tolu-enesulfonic acid monohydrate (92.8 mg, 488 CPmol) is added and the mixture stirred for 10 min at room temperature. The solvent is removed by rotary evaporation and the residue is triturated with ether (25 ml) to yield crude 107.

Fmoc-11-Aun-(carba-4-Cl-Ff)-R(Pbf)W-OMe (108) The procedure of making compound 104 is followed and crude 108 is obtained from Fmoc-11- Aun-OH (207 mg, 488 9mol), 107 (543 mg, 488 xmol), HOBt (66.0 mg, 488 Rmol), EDCI (103.0 mg, 537 gmol), and NMM (0.118 ml, 1.07 mmol).

S11-Aun-(carba-4-C1-Ff)-R(Pbf)W1 (109) 1 N NaOH (2.2 ml, 2.2 mmol) is added to a solution of 108 (520 mg, 387 ptmol) in THF-MeOH at room temperature and the resulting mixture is stirred at room temperature for 2 h. The mixture is acidified with 1 N HCI to pH ~3 and is partitioned between EtOAc (100 ml) and water ml). The water phase is further extracted with EtOAc (2 x 20 ml) and the combined organic phase is washed with brine (20 ml) and dried over MgSO 4 The dessicant is removed by filtration, the filtrate concentrated by rotary evaporation, and the residue triturated with ether ml) to the crude amino acid to be cyclized. NMM (47 1t, 460 ltmol) is added to a mixture of this crude amino acid (430 mg, 387 tmol), HOBt (53 mg, 387 itmol), and EDCI (82.5 mg, 430 Itmol), WO 02/26774 PCT/US01/30051 in DMF at 0°C. The resulting mixture is stirred at 0°C for 1 h and then at room temperature for 14 h. The mixture is worked-up as in the procedure for preparing compound 104 and purified by preparative HPLC to give 109.

S11-Aun-(carba-4-Cl-Ff)-RW1 (110) A mixture of 109 (42 mg, 39 gmol) in TFA-DCM-water (10:10:0.5, 5 ml) is stirred at room temperature for 20 h, the volatiles are removed by rotary evaporation, and the residue is purified by preparative HPLC. The product-containing fractions are combined, concentrated, frozen, and lyophilized to yield 110 as a TFA salt.

Examples 95-99 Coupling procedure (CP) for peptide bond formation in solution: An amine component (1 equivalent), an acid component (1 equivalent), and HOBt (2 equivalents) are dissolved in DMF (2 ml mmole of substrate). The solution is treated with Nmethylmorpholine (3-4 equivalents), EDCI (1.2 equivalent) and stirred at room temperature until formation of the product is complete (usually 1 5 hours). The product precipitates upon addition of water (6 10 ml ml DMF) to the reaction mixture and is separated from the liquid by filtration or decantation.

A. Synthesis of core dipeptide Boc-DPhe-Arg(N0 2 )OH (A-2) A-1. Boc-DPhe-Arg(N0 2 )OMe OgN HOBt O N ONH NH 0 NH DMF M NitJL H DMF HN 0 0 0 0 *2 O HN H o 0 HCI 0- 0 Reartant /iRea.nt M l±leq An'nt. 1Inito'; Boc-D-Phe 265.31 0.05 13.26 9 DMF _50 ml EDC 191.17 0.06 11.5 -T-Arf(OMel HCI 269.69 0.05 13.48g HOBt 13513 0.1 13.5 NMM j101.14 0.15 16.5 ml The coupling procedure (CP) for the peptide bond formation in solution is used.

98 WO 02/26774 WO 0226774PCT/US01/30051 A-2. Boc-DPhe-Arg(N0 2

)OH

LIOH

THF (A-2) flvetant t 'et Roc-DPhe-Arg(NWQhMe

TJIOH

TF

water MW! MalpeN: Amount., 2 T"IM!: 490.23 0.-01 4.R 8 23.95 0.023 0.55 10 ml A solution of Boc-DPhe-Arg(N02)OMe in THIF is cooled in an ice bath and treated at 0 0 C with aqueous solution of LiOH. The reaction mixture is stirred in the ice bath for 4 hrs.

Solvents are evaporated to small residual volume, which is treated with IN HG) (approx 25 ml) to pH 2-3. The product is extracted with ethyl acetate, washed with water brine, dried with anhydrous magnesium sulfate; the solvent is evaporated to dryness to afford the title compound.

B. Synthesis of amino terminus groups B-1. 3.(4-Benzyloxy-phenyl)-propionic acid N. Br 0

OH

HO" 1 NKOH EtDH 0

OH

Reacbqnit 1 Re~agent: n-hvdro~xvnroni onic acid henzvl hromide NaOH. 1iN FI01-

MW:

166.17 171.0.3 0.0468 0.0468 7.78 9-17 100 150 mrnl The procedure described in JACS 1955, 77, p. 4887 4892 is used. The product precipitates when the reaction mixture is acidified to pH 2 3.

99 WO 02/26774 WO 0226774PCT/US01/30051 B-2. 3-(4-Benzyloxy-phenyl)-propionyL chloride 0 0

PCI

OH CI 'N TOLUENE 3-(4-Benzyloxy-phenyl)-propionic 256.11 0.038 9.73 g acid PC] 95% 209.24 _D.0418 9.1.5 g toluene 400 ml Solid PCIS is added to a solution of 3-(4-benzyloxy-pheniyl)-propionic acid in toluene over a 1 hour period. The reaction mixture is stirred at room temperature for 3 hours and solvent is evaporated. The residue is stirred with hexanes overnight producing crystalline material, which is filtered and dried under vacuum.

B-3. 4-(S)-Benzyl-3-[3-(4-beiizyloxy-phenyl)-propionyl]-oxazolidin-2-one -O0 NH t-BuLi 0 NUi 0 0 0~ 'b A JN-- Reacbmnt tRepeynt: MW!, nk. Ammint: Vnits: be-n7vloxvnhenvlnronanovl chloride 274.74 0.005 1.37 g (S)-(--4-benzvlfoxazld n -177.2 -a.005 0-99 tart-RiJ~i in. nentane 0-0051 3" Ml TI-JR dry 6 M1 TT-TF. dry Iml The procedure described in Tetrahedron 52(43), 1996, pl3'733-l3738 is used. Li )-4-benzyloxazolidinone salt is prepared at -65' to -72'C. A solution of decanoyl chloride in THF is cooled to -72'C and treated with Li (S)-(-)-4-benzyloxazolidinone solution at this temperature. The reaction mixture is stirred at -70' to -75'C for 1 hour and overnight at room WO 02/26774 WO 0226774PCT/USOI/30051 temperature, treated with NII 4 Cl solution and extracted with ethyl acetate. The organic layer is washed with water/brine, dried with MgSO 4 and evaporated. The residue is separated on a silica column using hexane ethyl acetate 7 3 solution to afford the title compound.

B-4. 4-(S)-Benzyl-3-12-(4-bezyloxybenzyl)-pent-4-enoylj-oxazolidin-2-one NaHUMDS

THF

1Reactnnt I R.uiozent'! 1VflV Mn AmdnilinP Ilnigi 4-(S)-Benzyl-3-[3-(4-benizyloxy- 415.48 0.003 1.24 g phenyl)-propionyl]-oxazolidin-2one allyl bromide, d=1.398 120.98 0.006 0.52 ml TIHF 130 mlr NaHIVDS, 0.6 M in THF 5 mli NaTIMDS is added to a TIJF solution of 4-(S)-benzyl-3-113-(4-benzyloxy-phcnyl)propionyll-oxazolidin-2-one at -70' to -75'C over 15 min. The mixture is stirred for I hour and treated with allyl bromide at -70'C. Stirring is continued at this temperature for 1 hour. The reaction mixture is allowed to reach 0 0 C in 3 hours and quenched with 10% NH 4 Cl. The product is extracted with ethyl acetate, washed with water brine and dried with anhydrous magnesium sulfate. The solvent is evaporated and the crude produact is purified on a silica column using hexane 4 ethyl acetate 1 to afford the title compound.

2-(4-Bea2yloxy-benzyl)-pent-4-enoic acid WO 02/26774 WO 0226774PCT/USO1/30051

LICH

THE

TI-F I 1 m~ The procedure described in JOC 1992, 57(10), 2888-2902 (p.2894) is used.

B-6. 4-(4-(S)-Benzyl-2-oxo-oxazolidin-3-yl)-3-(4-beuzyloxy-benzyl)-4-oxo-butyronitrile 0 NaHMDS0 TH F

'CN

A procedure analogous to that used to prepare B-4 is used to prepare B-6.

B-7. 4-An'ino-2-(4-hydroxy-benzyl)-butyric acid ethyl ester; hydrochloride o)

OH

HO0 10% Pd/C 0 C CN EtOH MCI conc.0 N2H 2-Cyano~methyl-3-phenyl-propionic acid (8-16 g, 43 mmol) in a solution of ethanol ml) and concentrated HCI (10 ml) is hydrogenated overnight at 40 psi in the presence of WO 02/26774 WO 0226774PCT/USOI/30051 Pd/C. The catalyst is removed by filtration; the filtrate is concentrated under reduced pressure to dryness to afford the title compound.

B-8. 3-(4-Benzyloxy-phenyl)-2-decanoylaniino-propionic acid methyl ester 00

WN

0 0N 0~~ 0 N Y" H 0 Et3N

DOM

Rae itl"ti~nt,: H4-TV(13z,1)-OT\e HC1 decariovi chloride- 98% d=0.9 19 TEA. d=0.726i

DCM

321.9 0-003 0.965 190-71 006 1.7 m 101.19 0-016 2.2 MI 15 ml TEA is added to a solution of the remaining reactants in DCM at to +311C. The reaction mixture is stirred at room temperature for 4 brs and diluted with 0. IN IICI. The product is extracted with DCM, washed with water, dried With MgSO 4 and solvent is evaporated under reduced pressure. The residue is crystallized from hexanes to afford the title compound.

B-9. 3-(4-Benzyloxy-phenyl)-2-decanoylamiino-propionic acid N~ 1NNaOH N 0

THF

0 0 N N OH

H

0

H

H 0 0 Rqter 1 N NnOH water 1T Males- Amniint:' Units: 4-39. 59 a0022_3 0.98 .9 0003 2.3 n Ml n07 Ml Reactants are stirred at room temperature for 5 hirs. Solvents are evaporated under reduced pressure; the residue is diluted with water and acidified to pH about 2. Resulting WO 02/26774 PCT/US01/30051 precipitate of the product is filtered, washed with water until pH of the filtrate reaches about 6, and dried under vacuum overnight.

C. Synthesis of carboxyl terminal groups C-1. N-Benzyl-3-(1H-indol-3-yl)-N-methyl-propane-1,2-diamine HN HN

BH

3 .SMe2

H

2 N SMe H 2

N

0. THF O N N A 2 M THF solution of BH 3 Me 2 S complex (40 ml) is added to a solution of the amide substrate (4.5 g, 8.4 mmol) in anhydrous THF (50 ml). The reaction mixture is heated at while slowly distilling liquid is collected through the condenser. After 2 hours, a new portion of 2 M THF solution of BH 3 Me 2 S complex (10 iml) is added and heating with simultaneous distillation is continued for additional 3 hours. The reaction mixture is cooled to room temperature and carefully treated with MeOH, until the release of gas ceases, and with 3 N NaOH. The crude product is extracted with ethyl acetate and purified on a silica column using a solution of 1.5 MeOH in AcOEt following by a solution of EtOAc/DCM/MeOH/Et 3

N

4/5/0.5/0.3 to afford the title compopund.

C-2. N'-Benzyl-N-hexyl-3-(1H-indol-3-yl)-propane-1,2-diamine HN HN N ~BH 3 .SMe 2

H

2 N H 2

N

2 TFA THF A procedure analogous to that used to prepare C-1 was used to prepare C-2.

C-3. N-Hexyl-3-(1H-indol-3-yl)-N 1 -methyl-propane-1,2-diamine WO 02/26774 WO 0226774PCT/USOI/30051

HN

BH

3 SMe 2

THF

A procedure analogous to that used to prepare C-1 was used to prepare C-3.

D. Assembly of tetrapeptide mimetics ethylcarbamoyll-butylcarbamoyl I -2-:pheny-ethyl)-carbamic acid tert-butyl ester.

NH

HOBt, EDCI, NMM

DMF

Mu- N'MOR1~. 'E mcnin&! TI i cinontide 466.49 -0.0005 0.233.1__ N'-Hexyl-3-(1H-indol-3-yl)-N'- 287.24 0.0005 0.143 g methyl-propane-i ,2-dianiine_ H-ORt 135.12 0-001 0.135 9__ NMMd=0-92 101.14 0.0015.. 10.17 Iml EDCJ 19.7 0iX6 141 DM7F The coupling procedure (CP) for the peptide bond formation in solution is used. The crude product is purified on a silica column using hexane ethyl acetate 6 1 to afford the title compound.

A procedure analogous to that used to prepare D-1 is used to prepare the following D-2, D-3, D-4 and D-2. 2-f2-(2-tert-Butoxvcarbonylamino-3-phen poI L lamino)-S-nitro uanidinopentanoylaminol-3-( 1H-indol-3-yl)h-propionic acid methyl ester WO 02/26774 WO 0226774PCT/USO1/30051 HOBt, EDGI, NMM o A-2 DMF 61 D-3. (1-f 1-[2-(Benzyl-hexyl-anino)-1-(1H-indol-3-ylmethl)-ethylcarban-oli-4nitrogiuanidino-butylcarbamoyI I -2-phenvi-ethyD-carbamic acid tert-butyl ester HOBt, EDGI, NMM

DMF

D-4. 4-Nirosuanidino-l-r2-( 1f-indol-3-yl)-1-(mehvl-r Iv-carbamo )-ethvlcarbamoyll butvlcarbamoyl I 2-nhenvl-ethyfl-carbamic acid tert-butyl ester 0 2

N

N.,NH

2

NH

0 N0 NH H HO 0 2

N

N-yNH 2

NH

0

H

H NN 0 0 HOBt, EDGI, NMM

DMF

(1-f 1-[1-(Carbamoylmethvyl-methyl-carbamoyl)-2-(l1H-indol-3-yl)-ethylcarbamoyll -4nitroguanidino-butylcarbamoyl 1-2:phenyl-ethyi)-carbamnic acid tart-butyl este WO 02/26774 WO 0226774PCT/USOI/30051 NH HI HO HOBt, EDCI, NMM 0= Ozz<~ No O 002 ODMF

N-

A-2 I NH2 0 D-6. 2-(2-Amino-3- pheuyl-propio n-iino)-5-nitroguanidino-penitanoic acid r2-(hexYl-rneth3lamino)- 1H-indol-3-ylmethvl)-ethyll-aniide 0 2 N 0 2

N

NzyNH2

N-NH

NH N 0H 0H 0~ N 25% TFA/DCMH 2 0 0 N YM H> HHNHHN N-1

CF

0 COOH N TFA /DCM /H201/2/10.1 1 2 MI The reaction mixture is stirred at room temperature for 4 hours and diluted with 1,2dichloethane. Solvents are evaporated under reduced pressure; the residue is dried under vacuum overnight.

D-7. 2-(4-Benzyloxy-benzvl)-pent-4-enoic acid (l-i 4-nitroguanidino-1-r2-(hexy l-methyl-aiino)-1- (ll1-indol-3-vlmethl)-ethylcarbamoyll-butylearbamoyl 1-2-phen l-ethyl)-amide WO 02/26774 WO 0226774PCT/USOI/30051 02N NNH2

MH

o N

N

"H2 H HX HOBt

NMM

EDOI

DMF

RPearctnnI Rgea~ru: Ti k AaA~ni Mu-iis: Amide bond formation 149.92 0.000226 0.17 2cid 296.36 0.0003 0.09 H-O-Rt 135,12 0-0006 RV09 NMM- d=0.92 101.14 0.001 0.11 MI EDTCI .191.17 10.00036 M.069 .9 DMF 10.7 IMl The coupling procedure (CP) for the peptide bond formnation in solution is used. The crude product is purified by by reverse phase preparative HPLC to afford the title compound.

Example Synthesis of 2-(4-Hydroxy-benzyl)-pentanoic acid (1-{4-guanidino-1-[2-(hexyl-methylamino)- 1-(lH-indol-3-ylmethyl)-etlhylcarbamoyli-butylcarbamoyl}-2-phenyl-ethyl)-amide 0 2

N

NxNH2 HNz,,NH2 NH NH 0 H 0H N Pd(OH) 2 0 N NN I N N NH HHN C. AH H H o EOH HO/\ D-7 Reaebmnt./1eni'pnt! IM"ntes- Amriuiitt: TVnits IStnrfinp, material 913.15 01 Pt.0-T 115 MI Pd(OH4)2II The reaction mixture is hydrogenated at room temperature, 45 psi overnight. The catalyst is separated by filtration through Celite. Solvent is evaporated under reduced pressure. The crude product is purified by reverse phase preparative HPLC to afford the title compound.

Example 96 WO 02/26774 PCT/USOI/30051 Synthesis of Decanoic acid [1-(1-{4-guanidino-1-[2-(hexyl-methyl-amino)-1-(1H-indol-3yhnethyl)-ethylcarbamoyll-butylcarhamoyl-2-phenyl-ethylarbamoyl)-2-(4-hydroxyphenyl)-ethyl]-amide N0 2 N_ NH 2 0 OH0

NO

2

NR

Ho~l EM&l 0 H EDO

N

DM 0

H

HN, NI- 2 IHI N Pd(OI2

A/

OH

Reartnf llz ellt: TMW: Mls r!ni ~i~ acid 426-56 0-00023 01A ~mrne635.R 0.0001O9 DOJ2....

I4-RT 135.12 0-0005 (R07 NMM. d=0.92 101.14 0.001 0.11 ml FEDCT 1191.17 10003 10.06 19 1 DMF 1 ml] Pd(OH4)2 The coupling procedure (CP) for the peptide bond formation in solution is used.

Hydrogenation is performed in ethanol at 45 psi for 48 hrs. The catalyst is removed by filtration; the crude product is purified by reverse phase preparative TTPLC to afford the title compound.

A procedure analogous to that used to make Example 95 was used to prepare the compound of E xamples 97-99.

Example 97 Synthesis of 2-(5-Guanidino-2-{2-[2-(4-hydroxy-benzyl)-pentanoylamino]-3-phenylpropionylaniino}-pentanoylanino)3-(1H-indol-3-yl)-propiouic acid methyl ester WO 02/26774 WO 0226774PCT/USOI/30051

NH

2 HNzzzNH2 NH NH 0 H 0 H 0 N Pd(OH) 2 0D N NM NH HN /N 0 EtOH HO 0 Example 98 Synthesis of 2-(4-Hydroxy-benzyl)-pentanoic acid (1-{1-[1-(benzyl-methyl-carbamoyl)-2- (1H-indol-3-yl)-ethylearbamoyl]-4-guanidino-butylcarbamoyl}-2-phenyl-ethyl)-anide NH

N

[02 H 0 HN N Pd(OH) 2 HN H0 NH NH MeOH NH NH HO ON- 0Oc HN 00 Examle 99 Synthesis of [2-(1-1-[1-(Carbamoylrnethyl-methyl-carbamoyl)-2-(1H-indol-3-yl)ethylcarhamoyl]-4-guanidino-butylcarbamoyl}-2-phenyl-ethylcarbamoyl)-3-(4-hydroxyphenyl)-propyl]-carbamic acid tert-butyl ester

NH

NH

N02H 0a HN N N Pd(OH)2 H0 Y b 2 NH w N NH rH 2 N N

N

2 MeOH NI H L NH

+HN-

0-11- C. Manual Solid Phase Chemistry WO 02/26774 PCT/US01/30051 The following peptides are obtained by manual synthesis using Fmoc chemistry and Rink amide resin as the solid support. The removal of the Fmoc groups is achieved by reaction of piperidine in DMF for 30 min followed by washing with DMF (3 x 35 ml), MeOH (3 x 35 ml), and DMF (3 x 35 ml.) The ninhydrin color test is used for monitoring reaction completion.

Acetylation of the terminal amino group is done with 5% Ac 2 0/ 0.25 NMM/ 0.2% HOBt in DMF for 30 min followed by extensive washing with DMF and DCM and brief drying under vacuum. The crude product is cleaved from the resin and the protecting groups removed using 93% TFA and 2.3 ethanedithiol in water for 3 h at room temperature. After removal of the resin by filtration and washing with 3% TFA (3 x 18 ml), the filtrate is extracted with ether (6 x 20 ml) and is frozen and lyophilized. The crude product is then dissolved in 30% aqueous acetic acid and the solution purified by preparative HPLC (10t Vydac C 18 10 x 250 mm, 6 to acetonitrile-water TFA) gradient over 1.5 The product-containing fractions are combined and lyophilized to give the purified peptides.

Example 100 Synthesis of Ac-Y-(4-Py)ala-RW-NH2 2TFA Fmoc-W(Boc)-OH and Fmoc-R(pbf)-OH (each in 2-fold excess) are attached sequentially to the Rink amide resin (4.28g, 3 mmol) using PyBOP (2-fold excess) and NMM (4fold excess). After washing with DMF (3 x 35 ml), ether (4 x 35 ml), and drying in vacuo an increase in weight is achieved. The resin (1.17 g, 0.35 mmol) is suspended in DMF (10 ml), the Fmoc group is removed, PyBOP (0.6 g, 1.15 mmol), NMM (0.26 ml, 2.8 mmol), and Fmoc-(4- Py)ala-OH (0.447 g, 1.15 mmol) are added sequentially, and the mixture is shaken for 1 h.

Following Fmoc-deprotection, the coupling procedure is again repeated with Fmoc Y(t-Bu)-OH (0.528 g, 1.1 mmol). The peptide is then deprotected, acetylated, and cleaved from the resin to give of product.

Example 101 Synthesis of Ac-Y-(3-Py)ala-RW-NH2 2TFA Prepared according to Example 100, except Fmoc-(3-Py)ala-OH is used instead of Fmoc- (4-Py)ala-OH to yield the title compound.

WO 02/26774 PCT/US01/30051 Example 102 Synthesis of Ac-Y-(2-Py)ala-RW-NH2 2TFA Addition of Fmoc-(2-Py)ala-OH to a solution of PyBop and NMM in DMF leads to a very fast decomposition of the amino acid. Thus, this example is prepared using a modification of the procedure used for Example 100 in which NMM (88 ul, 0.8 mmol) is added to a solution of Fmoc-(2-Py)ala-OH (0.311 g, 0.8 mmol) and PyBrop (0.373 mg, 0.8 mmol). A second equivalent of NMM is then added after 15 min. After 100 min any un-reacted amino termini are acetylated and the remaining steps described in Example 100 are followed to give the title compound.

VIII. Composition and Method Examples Example A An obese human female subject weighing 130 kg is treated by this method to incur weight loss. Specifically, once each day for a period of 6 months, the subject is administered, via intravenous injection, 15 ml of an aqueous solution comprising the following: Component Concentration (mg/ml) Compound of Ex. 1 Sodium bisulfate 1 Sodium chloride 7 Chlorobutanol Citric acid Sterile water qs to 1 mL Sodium Hydroxide adjust to pH At the end of the treatment period, the patient exhibits measurable weight loss.

Example B An obese human male subject weighing 150 kg is subjected to a weight-reduction program that achieves weight loss with reduced adiposity through a combination of a restricted diet and increased exercise. Specifically, once each day for a period of 6 months after weight loss, the subject is administered, via intravenous injection, 15 ml of an aqueous solution comprising the following: Component Concentration (mg/ml) Compound of Ex. 9 Sodium bisulfate 1 WO 02/26774 PCT/US01/30051 Sodium chloride Chlorobutanol Citric acid Sterile water Sodium Hydroxide 7 qs to 1 mL adjust to pH At the end of the treatment period, the patient exhibits maintenance of weight loss and reduced adiposity.

Example C An obese human male subject weighing 165 kg is subjected to a weight reduction program that achieves weight loss through a combination of restricted diet, increased exercise and subcutaneous injection daily of 15 mls of an aqueous solution comprising: Component Compound of Ex. 88 Sodium bisulfate Sodium chloride Chlorobutanol Citric acid Sterile water Sodium Hydroxide Concentration (mg/ml) 1 7 qs to 1 mL adjust to pH Once the desired weight loss has been achieved, the patient's weight loss is maintained through continuation of the intravenous injection once each day for an additional period of 6 months. At the end of the treatment period the patient exhibits maintained weight loss and reduced adiposity.

Example D An obese human female subject weighing 140 kg is treated by the present method to incur weight loss. Specifically, she is treated with an implantable subcutaneous pump that delivers 0.1 mg/kg of the compound of Example 31 over a 24 hour period. The pump contains a solution of the compound dissolved in a solution of 50% propylene glycol and 50% sterile water.

The pump is replaced monthly and treatment continues for a six-month period at which time the patient exhibits weight loss and reduced adiposity.

Example E WO 02/26774 PCT/US01/30051 An obese male weighing 150 kg is treated by this method to incur weight loss.

Specifically, he is treated with an oral tablet taken twice daily containing 300 mg of the compound in Example 29. The treatment continues for 12 months at which time the patient exhibits weight loss and reduced adiposity.

Claims (27)

1. A compound having a structure according to Formula

2-C 2i l C 2 2C R (R R q(RR'(R R 11 2R wherein X is selected from hydrogen, fluoro, aryloxy, acyloxy, OR', SR', -NR 1 R 1 and -CHR'R", where R1 and R" are independently selected from the group consisting of hydrogen, alkyl and acyl; each R 2 is independently selected from the group consisting of hydrogen, alkyl halo and heteroalkyl; or two consecutive R moieties, or consecutive R 2 and W2 moieties, may join to form a 3 to 8 membered carbocyclic or heterocyclic ring; or the R 2 bonded to the carbon atom that is bonded to X and Z' and an R 5 moiety can optionally join to form a carbocyclic or heterocyclic ring that is fused to phienyl ring J; or the Rz bonded to the carbon atom that is bonded to ring Ar can join with an R7 to form a ring fused to ring Ar; or the R2 bonded to the carbon atom that is bonded to Z and Z3 can optionally join with R8 to form a carbocyclic or heterocyclic ring; or the R2 bonded to the carbon atom that is bonded to Z 3 and D can~optionally join with R 10 to form a carbocyclic or heterocyclic ring; each of e 2 and Z 3 is independently selected from -OC(R 3 )(R 3 -C(R 3 S(O).C(R 3 where a is 0, 1 or 2; -C(R 3 3 where b is 0, 1 or 2; N(R'e)C(R 3 -C(R 3 -C(O)C(R 3 C( 3 (R 3 ~CR)Ra)C(R 3 _SO 2 N(R 3 NRd)SO 2 _CR)RaP(=)Ol); -P(=O)(OR 3 f)C(R 3 _N(R 3 WO 02/26774 PCT/US01/30051 P(=O)(O 3 -O-P(=0)(OR 3 a cycloalkyl having from 3 to 8 ring atoms and a heterocycloalkyl having from 4 to 8 ring atoms; wherein each of R 3 R a R 3 b and R 3c when present, is independently selected from hydrogen, hydroxy, alkoxy, aryloxy, acyloxy, thiol, alkylthio, acylthio, arylthio, amino, alkylamino, acylamino, and alkyl; R 3d when present, is selected from hydrogen, alkyl and aryl; R 3 when present, is selected from hydrogen, alkyl, aryl and acyl; and R 3f when present, is selected from hydrogen and alkyl; p is 0, 1, 2, 3, 4 or 5; wherein when p is greater than 0, each R 4 and R 4 is independently selected from hydrogen, alkyl, aryl, halo, hydroxy, alkoxy, amino and acylamino; when p is greater than 1, two R 4 moieties, together with the carbon atoms to which they are bonded, can join to form a heterocycloalkyl, cycloalkyl or aryl ring; and when p is greater than 1, the R 4 moieties on two adjacent carbon atoms can both be nil such that a double bond is formed between the two adjacent carbon atoms, or both the R 4 and R 4 moieties on two adjacent carbon atoms can all be nil such that a triple bond is formed between the two adjacent carbon atoms; R 5 represents the 5 substituents positions 2-6) on phenyl ring J, wherein each R 5 is independently selected from hydrogen, hydroxy, halo, thiol, -OR 1 2 -SR 12 SO 2 N(R 2 )(R 1 2 -N(R 2 )(R 12 alkyl, acyl, alkene, alkyne, cyano, nitro, aryl, heteroaryl, cycloalkyl, and heterocycloalkyl; where each R 12 and R 1 2 is independently selected from hydrogen, alkyl, acyl, heteroalkyl, aryl, heteroaryl, cycloalkyl, and heterocycloalkyl; or two R 5 moieties can optionally join to form a carbocyclic or a heterocyclic ring that is fused to phenyl ring J; q is 0, 1, 2, 3, 4 or 5; wherein when q is greater than 0, each R 6 and R 6 is independently selected from hydrogen, alkyl, aryl, halo, hydroxy, alkoxy, amino and acylamino; when q is greater than 1, two R 6 moieties, together with the carbon atoms to which they are bonded, can join to form a heterocycloalkyl, cycloalkyl or aryl ring; and when q is greater than 1, the R 6 moieties on two adjacent carbon atoms can be nil such that a double bond is formed between the two adjacent carbon atoms, or both the R 6 and R 6 moieties on two adjacent carbon atoms can all be nil such that a triple bond is formed between the two adjacent carbon atoms; Ar is an aryl or heteroaryl ring selected from the group consisting of phenyl, thiophene, furan, oxazole, thiazole, pyrrole and pyridine; R 7 represents the substituents on ring Ar, wherein each R 7 is independently selected from hydrogen, halo, -NR 1 3 Ri', alkyl, acyl, alkene, alkyne, cyano, nitro, aryl, heteroaryl, cycloalkyl, and heterocycloalkyl; where each R 13 and R 1 3 is independently selected from hydrogen, alkyl, acyl, heteroalkyl, aryl, heteroaryl, cycloalkyl, and heterocycloalkyl; or two R 7 moieties can optionally join to form a carbocyclic or a heterocyclic ring fused to ring Ar; r is 0, 1, 2, 3, 4, 5, 6 or 7; wherein each R 8 and R 8 is independently selected from hydrogen, alkyl, halo, hydroxy, alkoxy and amino; when r is greater than 1, two R 8 moieties, together with the carbon atoms to which they are bonded, can join to form a heterocycloalkyl, cycloalkyl or aryl ring; and when r is greater than 1, the Ra moieties on two adjacent carbon atoms can be nil such that a double bond is formed between the two adjacent carbon atoms, or both the R' and R 8 moieties on two adjacent\arbon atoms can all be nil such that a triple bond is formed between the two adjacent carbon atoms; B is selected from -N(R 14 )C(=NR or 16R 1 7 -NR2°R21, cyano a heteroaryl ring eg. thiophene, an alkyl or dialkyl amme, a heteroaryl ring containing at least one ring nitrogen atom and a heterocycloalkyl ring containing at least one ring nitrogen atom, wherein R 1 4 R 1 5 R' 6 R i R 2 0 and R 21 are independently selected from hydrogen, alkyl, alkene, and alkyne; wherein further a combination of two or more of R 4 R 1 5 R 1 6 and R 17 may optionally combine with the atoms to which they are bonded to form a monocyclic or bicyclic ring; s is 0, 1, 2, 3, 4 or 5; wherein when s is greater than 0, each R 9 and R 9 is independently selected from hydrogen, alkyl, aryl, halo, hydroxy, alkoxy, amino and acylamino; when s is greater than 1, two R 9 moieties, together with the carbon atoms to which they are bonded, can join to form a heterocycloalkyl, cycloalkyl or aryl ring; and when s is greater than 1, the R 9 moieties on two adjacent carbon atoms can be nil such that a double bond is formed between the two adjacent carbon atoms, or both the R 9 and R 9 moieties on two adjacent carbon atoms can all be nil such that a triple bond is formed between the two adjacent carbon atoms; R'O is selected from the group consisting of an optionally substituted bicyclic aryl ring and an optionally substituted bicyclic heteroaryl ring; and D is independently selected from hydrogen, fluoro, hydroxy, thiol, acylthio, alkoxy, aryloxy, alkylthio, acyloxy, cyano, amino, acylamino, and wherein R" is selected from the group consisting of hydroxy; alkoxy; amino; alkylamino; NHOR' 8 where R' 8 is selected from hydrogen and alkyl; -N(R 19 )CH 2 C(O)NH 2 where R 1 9 is alkyl; -NHCH 2 CH20H;-N(CH 3 )CH 2 CH 2 OH; and -NHNHC(=Y)NH2, where Y is selected from O, S and NH; and wherein if at least one of Z 2 or Z 3 is other than -C(0)N(R 3 or -N(R 3 then X and D may optionally be linked together via a linking moiety, L, that contains all covalent bonds or covalent bonds and an ionic bond so as to form a cyclic peptide analog; or an optical isomer, diastereomer or enantiomer thereof; a pharmaceutically-acceptable salt, hydrate, or biohydrolyzable ester, amide or imide thereof. 2. A compound according to Claim 1 wherein B is selected from N(RI 4 )C(=NR 15 )NR 1 6 R' 7 cyano, N(R' 4 )C(=O)NR 16 R 7 a heteroaryl ring containing at least one ring nitrogen atom and a heterocycloalkyl ring containing at least one ring nitrogen atom.

3. A compound according to Claim 1 wherein B is selected from N(R' 4 )C(=NR' 5 )NR 1 6 R 17 -N(R' 4 )C(=O)NRI 6 R' 7 cyano, and trizole and imidazole.

4. The compound according to Claim 1 wherein X is selected from and CHR'R' and wherein R' is hydrogen or alkyl and R" is acyl.

5. The compound according to any one of Claims 1 to 4 wherein each R 2 is hydrogen; or the R 2 bonded to the carbon atom bonded to Z 3 and D joins with R 1 0 to form a carbocyclic or heterocyclic ring and the other R 2 moieties are hydrogen.

6. The compound according to any one of Claims 1 to 5, wherein Z 2 and Z 3 are independently selected from -OC(R 3 )(R 3 -C(R 3 )(R 3 -C(R 3 )(R 3 a)N(R 3 -C(O)N(R 3 C(R 3 )(R 3 a)C(R 3 b)(R 3 -C(R 3 )=C(R 3 -SO 2 N(R 3 and -P(=O)(OR)C(R 3 )(R 3 Y:ALoulse\P G\Spedes\687909_sped.doc

7. The compound according to Claim 6 wherein each R 3 R 3 a R 3 b, and R 3 c, when present, is independently selected from hydrogen, hydroxy, alkoxy, aryloxy, and alkyl; R 3 d, when present, is selected from hydrogen and alkyl; R 3 e when present, is selected from hydrogen and alkyl; and when R 3f is present and is alkyl, said R 3f is branched alkyl.

8. The compound of Claim 1 wherein p is 1 or 2.

9. The compound of Claim 1 wherein each R 4 when present, is hydrogen and each R 4 when present, is hydrogen or alkyl. The compound of Claim 1 wherein each R 5 is independently selected from hydrogen; hydroxy; halo; thiol; -SO 2 N(R12)(R 2 where R 1 2 and R 1 2 both are hydrogen; and -N(R 2 )(R 1 2 where R 12 and R 12 each are hydrogen or alkyl.

11. The compound of Claim 10 wherein four of the R 5 moieties on ring J are hydrogen.

12. The compound of Claim 10 wherein the 4-position of ring J is other than hydrogen.

13. The compound of Claim 1 wherein where q is greater than 0, each R 6 hydrogen and each R 6 is hydrogen or alkyl.

14. The compound of Claim 1 wherein q is 0, 1 or 2. The compound of Claim 1 wherein Ar is selected from the phenyl, thiophene and furan.

16. A compound of Claim 15 wherein Ar is phenyl and wherein, the 4-position of the phenyl ring is selected from hydrogen, fluoro, chloro, cyano, bromo, iodo, nitro and alkyl and the remaining four positions are hydrogen.

17. The compound of any one of Claims 1 to 16 wherein r is 2, 3 or 5 and each R 8 and R 8 is independently selected from hydrogen alkyl.

18. The compound of any one of Claims 1 or 4 to 17 wherein B is selected from N(R 14 )C(=NR' 5 )NR 1 6 R 1 7 a heteroaryl ring containing at least one ring nitrogen atom and a heterocycloalkyl ring containing at least one ring nitrogen atom.

19. The compound of Claim 18 wherein B is N(R1 4 )C(=NRI 5 )NR1 6 R 17 Y:\Louise\P G\Spedes\687909_spei.doc 119 The compound of Claim 18 or Claim 19 wherein R 14 R 15 R 16 and R 17 are independently selected from hydrogen and alkyl.

21. The compound of any one of Claims 1 to 20 wherein s is 1 or 2 and R 9 is hydrogen and each R 9 is hydrogen or alkyl.

22. The compound of any one of Claims 1 to 21 wherein R' 1 is selected from 1-naphthyl, 2- naphthyl, indan, 1H-indene, benzocyclobutane, benzocyclobutene, indole, indoline, pyrindine, dihydropyrindine, octahydropyrindine, benothiophene, benzofuran, benzimidozile, benzopyran, quinoline, quinolone and isoquinoline.

23. The compound of any one of Claims 1 to 22 wherein D is selected from fluoro, hydroxy, thiol, alkoxy, aryloxy, alkylthio, acyloxy, cyano, amino, acylamino, and C(S)R 1 1

24. The compound of any one of Claims 1 to 23 wherein X and D are linked together via linking moiety, L, to provide a cyclic compound having a structure according to the following Formula (II): A compound having a structure according to Formula SR 2 R\ N-C--Z R I Y:\Louise\P G\Spedes\687900_sped.doc wherein R 1 and R" are independently selected from the group consisting of hydrogen, alkyl and acyl; R 2 is selected from the group consisting of hydrogen, alkyl and heteroalkyl; Z' is selected from -C(R 3 )(R 3 -S(O) 2 C(Ri)(Ria); -C(Ri)(R 3a)S(O)2- N(Rie)C(Ri)(Ra)-; -C(Ri3)(Ra)N(Re)-; -N(Ri3d)C(O)_; C(Ri)(Ria)C(RIb)(R)-; -SO2N(Rild)_. _N(Rld)SO 2 C(R 3 3 and 3)(R wherein each of R 3 R 3 a Rib and R3, when present, is independently selected from hydrogen, hydroxy, alkoxy, aryloxy, acyloxy, thiol, allcylthio, acylthio, arylthio, amino, alkylamino, acylammno, and alkyl; (2 Miwe rsni eetd rmhdoealy n rl Rio, when present, is selected from hydrogen, alkylan aryl da;an R 3 when present, is selected from hydrogen, lyay and a yl; an p f is1 hr n peahRnd is iidpndnl selected from hydrogen, aanyd arykhalo (D hysd rakoxy, R amn and acylasinoeednl;eetdfo ydoeakl rl ao Risslefrmhydrogen, hydoxy, amionfor, NRb( 1 )whr and R 12 'mno each are independently selected from hydrogen and alkyl; q is 0, 1 or 2; wherein when q is greater than 0, each R 6and R 6 is independently selected from hydrogen, alkyl, aryl, halo, hydroxy, alkoxy, amino and acylainino; :R 7 is selected from hydrogen, halo, -NR 13R 13, alkyl, acyl, alkene, alkyne, cyano, nitro, aryl, heteroaryl, cycloalkyl, and heterocycloalkyl; wherein each R" 3 and R 13 is independently selected from hydrogen and alkyl; B is selected from -N(R' 4 )c(=NR' 5 )NR1 6 RI 7 a heteroaryl ring containing at least one ring nitrogen atom and a heterocycloalIcyl ring containing at least one ring nitrogen atom; wherein R 14 R' 5 R" 6 and R 1 7 are independently selected from hydrogen and alkyl; wherein furthier a combination of two or more of R 14 R 1 5 R'1 6 and R 1 7 may optionally combine with the atoms to which they are bonded to form a monocyclic or bicyclic ring; R1 0 an optionally substituted bicycdic ring selected from 1 -naphthyl, 2-naphthyl, indan, IH-indene, benzocylcobutane, benzocylcobutene, indole, indoline, pyrindine, dihydropyrindine, octahydropyrindine, benzothiophene, benzofuran, benzimidozole, benzopyran, quinoline, quinolone and isoquinoline; and wherein R 14 R 1 5 R 16 and R 1 7 are independently selected from hydrogen and alkyl; wherein further a combination of two or more of R' 4 R s 1 R 16 and R 17 may optionally combine with the atoms to which they are bonded to form a monocyclic or bicyclic ring; R'O an optionally substituted bicyclic ring selected from 1-naphthyl, 2-naphthyl, indan, 1H-indene, benzocylcobutane, benzocyclobutene, indole, indoline, pyrindine, dihydropyrindine, octahydropyrindine, benzothiphene, benzofuran, benzimidozole, benzopyran, quinoline, quinolone and isoquinolone; and R" is selected from the group consisting of amino; alkylamino; -NHOR 1 8 where R' 8 is selected from hydrogen and alkyl; -N(R' 9 )CH 2 C(O)NH 2 where R 9 is alkyl; NHCH 2 CH20H; and -N(CH 3 )CH 2 or an optical isomer, diastereomer or enantiomer thereof; a pharmaceutically-acceptable salt, hydrate, or biohydrolyzable ester, amide or imide thereof.

26. A pharmaceutical composition comprising: a safe and effective amount of a cyclic peptide anolog of any one of Claims 1 to 25 and a pharmaceutically-acceptable excipient.

27. A pharmaceutical composition of Claim 26 to be used in treating a disorder selected from the group consisting of insulin resistance, glucose intolerance, Type-2 diabetes mellitus, coronary artery disease, elevated blood pressure, hypertension, dyslipidaemia, cancer endometrial, cervical, ovarian, breast, prostate, gallbladder, colon), menstrual irregularities, hirsutism, infertility, gallbladder disease, restrictive lung disease, sleep apnea, gout, osteoarthritis, and thromboembolic disease in an animal subject.

28. A pharmaceutical composition of Claim 27 to be used in treating a body weight disorder selected from the group consisting of obesity, anorexia, and chachexia in an animal subject.

29. A method of treatment or prophylaxis of a disorder selected from the group consisting of insulin resistance, glucose intolerance, Type-2 diabetes mellitus, coronary artery disease, elevated blood pressure, hypertension, dyslipidaemia, cancer endometrial, cervical, ovarian, breast, prostate, gallbladder, colon), menstrual irregularities, hirsutism, infertility, gallbladder disease, restrictive lung disease, sleep apnea, gout, osteoarthritis, and thromboembolic disease, said method comprising use of a pharmaceutical composition according to Claim 27. Y:\Louse\P G\Spedes\87009_spedoc I A method of treatment or prophylaxis of a body weight disorder selected from the group consisting of obesity, anorexia, and chachexia in an animal subject, said method comprising use of a pharmaceutical composition according to Claim 27.

31. A compound according to Claim 1 substantially as hereinbefore described with reference to the examples.

32. A compound according to Claim 25 substantially as hereinbefore described with reference to the examples. Dated: 22 April 2005 PHILLIPS ORMONDE FITZPATRICK Attorneys for: THE PROCTER GAMBLE COMPANY Y:\LoulseP GSpedes\687909_sped.doc