IE921321A1 - Ó-substituted polypeptides having therapeutic activity - Google Patents

Abstract

The compounds of the invention are alpha -substituted mono-, di-, tri-, tetra-, and pentapeptides useful in treating obesity, anxiety, gastrointestinal ulcers, pain, stroke, and inflammation. They are also useful in blocking the reaction caused by withdrawal from drug or alcohol use and in reducing gastric acid secretion. They are further useful as agents in hypertension, heart failure, stroke, cognition, memory enhancement, spasticity, depression, diabetes, cancer, asthma, bladder dysfunction, psychosis, and arthritis and/or inflammatory pain. Pharmaceutical compositions, novel intermediates, and processes are also included.

Description

Peptides form the main messenger systems within and between cells and they number more than a thousand. Over a hundred peptides are known to act as hormone, neurohormones, or neurotransmitters, and this number is growing rapidly. The potential for drug development is therefore vast. However, the great majority of peptide messengers are not suitable for use as pharmaceuticals in their natural state.

The problems of natural peptides as drugs are lack of oral activity, failure to penetrate the blood-brain barrier, rapidly metabolized, no selectivity for receptor subclasses, antigenic properties, and expensive to make.

The vast majority of small peptide messengers are not suitable as drugs, particularly where an orally administered and possibly centrally active drug is required. In this situation the development of modified peptides offer significant opportunities.

The a,a-disubstituted amino acids are non-genetically coded synthetic analogues of natural mammalian α-amino acids and are incorporated at least once into the neuropeptides of this patent. -2Some examples of therapeutic applications of these modified peptides (peptoids) are given in Table I below.

TABLE I Endogenous Peptide Peptoid Type* Therapy Angiotensin ANT Hype rtension Heart Failure Atrial Natriuretic Factor AG Heart Failure Thyrotropin Releasing Factor AG Stroke Cognition Spasticity Depression Neuropeptide-Y ANT Hypertension Depression Obesity Glucagon ANT Diabetes Insulin AG Diabetes Gastrin ANT Gastric Ulcer Cancer Bombesin ANT Cancer Tachykinins SP, NKA, NKB ANT Antipsychotic Analgesic Antiinflammatory Antiasthmatic * ANT = Antagonist; AG = Agonist These offer completely novel approaches to drug treatment. For example, all of the major tranquilizers block central dopaminergic function indiscriminately. An antipsychotic modified peptide designed to act through the mesolimbic CCK-neuropeptide/Dopamine system may be considerably more selective. An improvement in quality through the ability to modify drug resistant characteristics -3is also expected. In psychosis the blunting of affect leading to general impoverishment of social interactions with schizophrenics is expected to be susceptible to alternate modified peptide therapies.

The highly selective behavioral responses elicited by individual neuropeptides is shown in the Table II below.

TABLE II Behavioral Responses to Peptides Peptide Behavior Angiotensin-II Dipsogenesis Cholecystokinin Cessation of Feeding Drowsiness Enhanced Memory Adrenocorticotrophic Hormone Enhanced Alertness Enhanced Cognition β-Endorphin Decreased Awareness Amnesia Reward Analgesia Dynorphin Decreased Reward Drowsiness Analgesia Luteinizing Hormone Releasing Factor Increased Sexual Receptivity Corticotrophin Releasing Factor Anxiety Enhanced Vigilance Thyrotropin Releasing Factor Increased Activity Enhanced Awareness SP or NKA A dopamine behavioral syndrome, i.e., increased locomotion, rearing SP Antidipsogenic activity -4SUMMARY OF THE INVENTION The invention relates to novel compounds of formula R2 I 3 R-NH-C-COR I u R and the pharmaceutically acceptable salts thereof 12 3 4 wherein R , R , R , and R are as defined hereinbelow.

In commonly assigned copending application 07/609,754, filed on April 24, 1991, by Horwell, et al, the disclosure of which is incorporated by reference, CCK analogues containing α,α-disubstituted amino acids are disclosed.

The invention also relates to a pharmaceutical composition containing an effective amount of a compound according to formula I in combination with a pharmaceutically acceptable carrier in unit dosage form for appetite suppression. The invention also relates to a method for suppressing appetite in a mammal.

The compounds of the invention are also useful for blocking the reaction caused by withdrawal from drug or alcohol use. The compounds of the invention are also useful in reducing gastric acid secretion, in treating gastrointestinal ulcers, in treating pain, treating and/or preventing stroke, treating inflammation, and in treating anxiety.

The compounds of the invention are also useful in treating cognitive deficits, small cell lung cancer, colonic cancer, peptic ulcers, and are useful in contraception. -5The invention also relates to a pharmaceutical composition for reducing gastric acid secretion containing an effective amount of a compound of formula I in combination with a pharmaceutically acceptable carrier in unit dosage form effective for reducing gastric acid secretion.

The invention also relates to a method for reducing gastric acid secretion in mammals which comprises administering an amount effective for gastric acid secretion reduction of the composition described above to a mammal in need of such treatment.

The invention also relates to a pharmaceutical composition containing an effective amount of a compound of formula I in combination with a pharmaceutically acceptable carrier in unit dosage form effective for reducing anxiety.

The invention also relates to a method for reducing anxiety in mammals which comprises administering an amount effective for anxiety reduction of the composition described above to a mammal in need of such treatment.

The invention also relates to a pharmaceutical composition containing an effective amount of a compound of formula I in combination with a pharmaceutically acceptable carrier in unit dosage form effective for treating gastrointestinal ulcers.

The invention further relates to a method for treating gastrointestinal ulcers in mammals which comprises administering an amount effective for gastrointestinal ulcer treatment of the composition as described above to a mammal in need of such treatment.

The invention also relates to a pharmaceutical composition containing an effective amount of a -6compound of formula I in combination with a pharmaceutically acceptable carrier in unit dosage form effective for treating inflammation.

The invention further relates to a method for 5 treating inflammation in mammals which comprises administering an amount effective of a composition as described above to a mammal in need of such treatment.

The invention also relates to a pharmaceutical 10 composition for preventing the withdrawal response produced by chronic treatment or abuse of drugs or alcohol.

The invention further relates to a method for treating the withdrawal response produced by withdrawal from chronic treatment or withdrawal from abuse of drugs or alcohol. Such drugs include benzodiazepines, especially diazepam, cocaine, caffeine, opioids, alcohol, and nicotine. Withdrawal symptoms are treated by administration of an effective withdrawal treating amount of a compound of the instant invention.

This invention also relates to a pharmaceutical composition containing a therapeutically effective amount of a compound according to formula I in combination with a pharmaceutically acceptable carrier in unit dosage form for treating psychosis.

The invention also relates to a method for treating psychosis in mammals which comprises administering an amount effective for treatment of the composition described above to a mammal in need of such treatment.

This invention also relates to a pharmaceutical composition containing a therapeutically effective amount of a compound according to formula I in —7 — combination with a pharmaceutically acceptable carrier in unit dosage form for treating asthma.

The invention also relates to a method for treating asthma in mammals which comprises administering an amount effective for treatment of the composition described above to a mammal in need of such treatment.

This invention also relates to a pharmaceutical composition containing a therapeutically effective amount of a compound according to formula I in combination with a pharmaceutically acceptable carrier in unit dosage form for treating bladder dys function.

The invention also relates to a method for treating bladder dysfunction in mammals which comprises administering an amount effective for treatment of the composition described above to a mammal in need of such treatment.

This invention also relates to a pharmaceutical composition containing a therapeutically effective amount of a compound according to formula I in combination with a pharmaceutically acceptable carrier in unit dosage form for treating arthritis and/or inflammatory pain.

The invention also relates to a method for treating arthritis and/or inflammatory pain in mammals which comprises administering an amount effective for treatment of the composition described above to a mammal in need of such treatment.

The invention further relates to methods of treating hypertension, heart failure, stroke, cognition, memory enhancement, spasticity, depression, and diabetes.

The invention further provides processes for the preparation of compounds of formula I. -8The invention further provides novel intermediates useful in the preparation of compounds of formula I and also provides processes for the preparation of the intermediates.

The invention also relates to a pharmaceutical composition for treating pain and to a method of using a compound of formula I for treating pain.

The invention also relates to a pharmaceutical composition for treating and/or preventing stroke and to a method of using a compound of formula I for treating and/or preventing stroke.

DETAILED DESCRIPTION The following provides a dictionary of the terms used in the description of the invention.

Abb revi at i on Side Chain Amino Acid ALA ch3- NH II NH2-C-NH-CH2-CH2-CH2- Alanine ARG Arginine ASN 0 II NH2-C-CH2- Asparagine ASP hooc-ch2- Aspartic Acid CYS hs-ch2- Cysteine GLU HOOC-CH2-CH2- Glutamic Acid O II GLN NH2-C-CH2-CH2Glutamine Abb reviation Side Chain Amino Acid HIS /“V-ch2— V Histidine ILE fH3 CH3-CH2-CH- Isoleucine LEU CHO-CH-CHO- 3 1 2 ch3 Leucine 5 LYS NH2-CH2-CH2-CH2-CH2- Lysine MET CH3-S-CH2“CH2- Methionine PHE ch2— Phenylalanine PRO SER THR H2C-(N“) h2C'c/ h2 HO-CH2TYR CH2 — ίι -iT“CH2' Proline Serine Threonine Tyrosine TRP Tryptophan -10Abbreviation Side Chain Amino Acid VAL CH--CH3 I ch3 Valine Abb revi at ion N-Terminal Protecting Group H Hydrogen BOC Tert-butvloxy- carbonyl CBZ Benzyloxy- (or Z) carbonyl IBU Isobutyryl IVA Isovaleryl NVA n-Valeryl 1-Adoc 1-Adamanty1oxycarbonyl 2-Adoc 2-Adamantyloxycarbonyl Fmoc 9-Fluorenyl- methoxycarbonyl C-Terminal Substituent/ Side Chain Protecting Abbreviation Group -nh2 Amide -OCH3 Methyl ester C-Terminal Substituent/ Side Chain Protecting Group Benzyl ester/ether Tert-butyl ester/ether -11Abbreviation -OCH2Ph -OC(CH3)3 The term N-terminal protecting as used herein refers to those groups known to the art intended to protect the N-terminus of an amino acid or peptide or to protect an amino group against undesirable reactions during a synthetic procedure or to prevent the attack of exopeptidases on the compounds or to increase the solubility of the compounds and includes, but is not limited to, sulfonyl, acetyl, pivaloyl, t-butyloxycarbonyl (Boc), benzyloxycarbonyl (Cbz), benzoyl, or an L- or D-aminoacyl residue, which may itself be N-protected similarly.

The term C-terminal protecting group as used herein refers to those groups known to the art intended to protect the C—terminus of an amino acid or peptide, these include but are not limited to an amide, methyl ester, benzyl ester/ether, tert-butyl ester/ether. Other examples of side chains and protecting groups are those known in the art. Any of those could be used.

The compounds of the invention are represented by formula -12R2 R1-NH-C-COR3 and the pharmaceutically acceptable salts thereof, wherein r1 is an N-terminal blocking group or from 0 to 4 amino acid residues or hydrogen; R is a sidechain of a genetically coded amino acid except glycine; R is a C-terminal blocking group or from 0 to amino acid residues, or -OH, or 0Rn where Rn is straight or branched alkyl or cycloalkyl of 1 to 6 carbon atoms; R is a sidechain of a genetically coded amino acid, except glycine, or -hc=ch2, —C=CH, -ch2-ch=ch2, -ch2c=ch, -CH2Ar, -ch2or, -CH2OAr, - (CH2)nCO2R, -(CH2)nNR5R6 wherein n is an integer of from 0 to 3, R is hydrogen or lower alkyl, Ar is a mono- or polycyclic unsubstituted or substituted carbo- or heterocyclic aromatic or hydroaromatic moiety; neither R2 nor R4 can be hydrogen; R1 plus R3 cannot be greater than 4 amino acid residues in total. -13Preferred compounds of the invention are those of formula I selected from: Formyl—MeMet—Leu—Phe—Obzl, F o rmy1-Met-MeLeu—Phe-OBz 1, Formyl-Met-Leu—MePhe—OBzl, Ac-MeLeu-Leu-Arginal, Ac-Leu-MeLeu-Arginal, MeTyr-Arg, Tyr—MeArg, MeTyr-Pro-Phe-Pro-NH2, Tyr-MePro-Phe—Pro-NH2, Tyr-Pro—MePhe-Pro-NH2, Tyr-Pro—Phe-MePro—NH2, Me-Arg-Lys-Asp-Val—Tyr, Arg-MeLys-Asp-Val-Tyr, Arg-Lys-MeAsp-Val-Tyr, Arg-Lys-Asp-MeVal-Tyr, Arg-Lys-Asp-Val-MeTyr, MeArg-Lys-Glu-Val-Tyr, Arg-MeLys—Glu-Val-Tyr, Arg-Lys—MeGlu-Val-Tyr, Arg-Lys—Glu-MeVal-Tyr, Arg-Lys-Glu-Val-MeTyr, MeAsp-Leu-Asp-Pro-Arg, Asp-MeLeu-Asp-Pro-Arg, Asp-Leu-MeAsp-Pro-Arg, Asp-Leu-Asp-MePro-Arg, Asp—Leu—Asp-P ro—MeArg, MeLys-Trp-Lys, Lys-MeTrp-Lys, Lys-Trp-MeLys, MePhe—Met—Arg—Phe-NH2, Phe-MeMet-Arg-Phe-NH2, Phe-Met-MeArg-Phe-NH2, Phe-Met-Arg-MePhe-NH2, -14MeGlp-His-Pro, Glp-MeHi s-P ro, Glp-His-MePro, MeArg-Tyr-Leu-Prο-Thr, Arg—MeTyr-Leu-Pro—Thr, Arg—Tyr-MeLeu—ProThr, Arg-Tyr-Leu-MePro-Thr, Arg-Tyr-Leu-Pro-MeThr, MeThr-Lys-Pro-Arg, Thr-MeLys-Pro-Arg, Th r-Ly s-MeP ro-Arg, Thr—Lys—Pro—MeArg, MeThr-Pro-Arg-Lys, Thr-MePro-Arg-Lys, Thr-Pro-MeArg-Lys, Thr—Pro-Arg-MeLys, MeThr—Val-Leu, Th r-MeV a1-Leu, and Thr-Val-MeLeu.

More preferred compounds of the invention are those of formula I selected from: Me-Tyr—Gly-Gly-Phe—Met, MeTyr—Gly—Gly—Phe—Met—NH2, MeTyr-Gly—Gly-Phe—Leu—NH2, Tyr—Gly-Gly—MePhe—Met, Tyr-Gly-Gly-MePhe—Met—NH2, Tyr-Gly-Gly-MePhe—Leu—NH2, Tyr-Gly-Gly-Phe—MeMet, Tyr—Gly—Gly—Phe—MeMet—NH2, Tyr—Gly—Gly—Phe—MeLeu—NH2, Z-DMePhe-Phe—Gly, Z-DPhe—MePhe—Gly, MeArg-Pro-Tyr-Ile-Leu, Arg-MePro-Tyr-Ile—Leu, Arg-Pro-MeTyr-Ile—Leu, -15Arg-Pro-Tyr-Melle-Leu, Arg-Pro-Tyr-Ile-MeLeu, MeLeu-Asp-Ile—Ile-Trp, Leu-MeAsp-Ile-Ile-Trp, Leu-Asp-Melle-Ile-Trp, Leu-Asp-Ile-Melle-Trp, Leu-Asp-Ile-Ile-MeTrp, MeGlu-Cys-Tyr—Phe, Glu-MeCys-Val-Tyr-Phe, Glu-Cys-MeVal-Tyr-Phe, Glu-Cys-Val-MeTyr-Phe, and Glu-Cys-Val-Tyr-MePhe.

Most preferred compounds of the invention are those of formula I selected from: MeLys-Trp-Asp-Asn-Gln, Lys-MeTrp-Asp-Asn—Gin, Lys-Trp-MeAsp-Asn—Gin, Lys-Trp-Asp-MeAsn-Gln, Lys-Trp-Asp-Asn-MeGln, MeVal-Gly-His-Leu—Met-NH2, Val-Gly-MeHis—Leu—Met-NH2, Val-Gly—His—MeLeu—Met—NH2, Val-Gly-His-Leu-MeMet-NH2, MeGlp-His-Trp—Ser-Tyr, Glp-MeHis-Trp-Ser-Tyr, Glp-His-MeTrp-Ser-Tyr, Glp-His-Trp-MeSer-Tyr, Glp-His—Trp—Ser—MeTyr, Gly-MeLeu-Arg-Pro-Gly—NH2, Gly-Leu-MeArg-Pro-Gly-NH2, Gly-Leu-Arg-MePro-Gly-NH2, MeTyr-Pro-Ser-Lys-Pro, Tyr—MePro-Ser—Lys—Pro, Tyr—Pro—MeSer—Lys—Pro, Tyr-Pro—Ser-MeLys-Pro, -16Tyr-Pro-Ser-Lys-MePro, MeThr-Arg-Gln-Arg-Tyr-NH2, Thr-MeArg-Gln-Arg-Tyr—NH2, Thr-Arg-MeGln-Arg-Tyr-NH2, Thr-Arg-Gln-MeArg-Tyr-NH2, Thr-Arg-Gln-Arg-MeTyr-NH2 r Gly-MeTrp-Thr-Leu-Asn, Gly-Trp-MeThr-Leu—Asn, Gly-Trp-Thr-MeLeu-Asn, Gly-Trp-Thr-Leu-MeAsn, MeLeu-Tyr-Gly-Leu-Ala-NH2, Leu—MeTyr—Gly—Leu—Ala—NH2, Leu-Tyr—Gly—MeLeu—Ala—NH2, Leu-Tyr-Gly-Leu-Aib-NH2, MePhe-Phe-Trp-Lys-Thr, Phe-MePhe-Trp-Lys-Thr, Phe-Phe-MeTrp-Lys—Thr, Phe-Phe-Trp-MeLys-Thr, Phe-Phe-Trp-Lys—MeThr, 0 MePhe-Phe-Gly-Leu—Met-NH2, Phe-MePhe-Gly-Leu-Met-NH2, Phe-Phe-Gly-MeLeu-Met-NH2, Phe-Phe-Gly-Leu-MeMet-NH2, and the Na-4-hydroxyphenylacetyl derivatives and the NH2—CO—(CH2)4—CO— derivatives of the above four compounds, N-[[(4-Chiorophenyl)methoxy]carbonyl]—L— t rypt ophy1-α-methy1-DL—pheny1a1anin amide, N-[[[4-(Trifluoromethyl)phenyl]methoxy]30 carbonyl]-L-tryptophyl-a-methyl-DL-phenylalaninamide, N-[([1,1'-Biphenyl]-4-ylmethoxy)carbonyl]L-tryptophvl-g-methyl-DL-phenylalaninamide, N—[(9-Anthracenylmethoxy)carbonyl]—L—tryptophy1g-methyl-DL-phenvlalaninamide, -17N-[(1-Naphthalenylmethoxy)carbonyl]-Lt ryptophy1-α-methy1—DL-pheny1a1aninami de, N-[(1-Naphthalenylmethoxy)carbonyl]-Lt rypt ophy1-α-methy1-L-phenylalaninamide, N-[(1-Naphthalenylmethoxy)carbonyl]-Lt rypt ophy1-α-methyl-D-pheny1a1aninamide, N-[[[4-(Propoxycarbonyl)phenyl]methoxy] carbonyl] —L—tryptophyl-Ct—methyl-DLphenylalaninamide, N-[(Phenylmethoxy)carbonyl]-L-tryptophy1-amethyl-DL-phenylalanylglycinamide, N-[(1-Naphthalenylmethoxy)carbonyl]-Ltryptophyl-a-methyl-DL-phenylalanylglycinamide, N-[(1-Naphthalenylmethoxy)carbonyl]-L15 tryptophyΙ-α-methyl-L-phenylalanylglycinamide, and N-[[(2,3-Dimethoxyphenyl)methoxy]carbonyl]L-t rypt ophy1-α-methy1-DL—phenylalaninamide .

The compounds include solvates, hydrates, and pharmaceutically acceptable salts of the compounds of formula I above.

The compounds of the present invention may exist as diastereomers, mixtures of diastereomers, or as the mixed or the individual optical enantiomers. The present invention contemplates all such forms of the compounds. The mixtures of diastereomers are typically obtained as a result of the reactions described more fully below. Individual diastereomers may be separated from mixtures of the diastereomers by conventional techniques such as column chromatography or repetitive recrystallizations.

Individual enantiomers may be separated by conventional methods well known in the art such as conversion to a salt with an optically active compound, followed by separation by chromatography or -18recrystailization and reconversion to the nonsalt form.

The compounds of the present invention may be formed by coupling individual substituted a-amino acids by methods well known in the art. (See, for example, standard synthetic methods discussed in the multi-volume treatise The Peptides, Analysis, Synthesis, Biology, by Gross and Meienhofer, Academic Press, New York). If known, the individual substituted alpha amino acid starting materials are synthesized by methods within the skill of the art. (Synthesized racemic [DL]-α-methyl tryptophan methyl ester - see Brana, M. F., et al, J. Heterocyclic Chem., 1980.

In Scheme I below, the α-MePhe was N-terminally protected with (BOC)2O to afford BOC a—MePhe.(1) Compound 1 was then activated with DCCI and HOBt and reacted with NH3 (g) to give the amide. (2) Removal of the BOC group with TFA gave the amine (3), which was reacted with FMOC-L-TrpOPfp to afford the dipeptide.(4) A further deprotection with piperidine in DMF gave the amine (5), which was reacted with the appropriate chloroformate to give the Examples 7 through 14.(6-13) Ph H2N*^CO2H D, L or D or L -19SCHEME 1 (Examples 2-13) Ph L^ch3 o2cn'4^co2h H conh2 H CH3 ROCOHN & - 12 (Examples 7-14) -20In Scheme 2 below, Compound 1 was activated with DCCI and HODhbt to give 14, which was reacted with CTlyNH2 in EtOAc to afford the N-protected dipeptide.(15) Removal of the BOC group with TFA gave the amine (16), which was reacted with ROCO(L) Trp activated with HBTU to give the Examples 18 and 19. (17,18) -21SCHEME 2 (Examples 2, 14, and 16-19) Ph --o2cn'^co,h H Ph L^H3 h2n^xconh if conh2 12, lfi (Examples 18 and 19) Ph k<CH3 .

O2CN CONH CONH2 H -22For preparing pharmaceutical compositions from the compounds of this invention, inert, pharmaceutically acceptable carriers can be either solid or liquid. Solid form preparations include powders, tablets, dispersible granules, capsules, cachets, and suppositories.

A solid carrier can be one or more substances which may also act as diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders, or tablet disintegrating agents; it can also be an encapsulating material.

In powders, the carrier is a finely divided solid which is in a mixture with the finely divided active component. In tablets, the active component is mixed with the carrier having the necessary binding properties in suitable proportions and compacted in the shape and size desired.

For preparing suppository preparations, a low-melting wax such as a mixture of fatty acid glycerides and cocoa butter is first melted and the active ingredient is dispersed therein by, for example, stirring. The molten homogeneous mixture is then poured into convenient sized molds and allowed to cool and solidify.

The powders and tablets preferably contain 5% to about 70% of the active component. Suitable carriers are magnesium carbonate, magnesium stearate, talc, lactose, sugar, pectin, dextrin, starch, tragacanth, methyl cellulose, sodium carboxymethyl cellulose, a low-melting wax, cocoa butter, and the like.

The term preparation is intended to include the formulation of the active component with encapsulating material as a carrier providing a capsule in which the active component (with or without other carriers) is surrounded by a carrier -23which is thus in association with it. Similarly, cachets are included.

Tablets, powders, cachets, and capsules can be used as solid dosage forms suitable for oral administration.

Liquid form preparations include solutions, suspensions, and emulsions. Sterile water or water-propylene glycol solutions of the active compounds may be mentioned as an example of liquid preparations suitable for parenteral administration.

Liquid preparations can also be formulated in solution in aqueous polyethylene glycol solution.

Aqueous solutions for oral administration can be prepared by dissolving the active component in water and adding suitable colorants, flavoring agents, stabilizers, and thickening agents as desired.

Aqueous suspensions for oral use can be made by dispersing the finely divided active component in water together with a viscous material such as natural synthetic gums, resins, methyl cellulose, sodium carboxymethyl cellulose, and other suspending agents known to the pharmaceutical formulation art.

Preferably the pharmaceutical preparation is in unit dosage form. In such form, the preparation is in unit dosage form. In such form, the preparation is divided into unit doses containing appropriate quantities of the active component. The unit dosage form can be a packaged preparation, the package containing discrete quantities of the preparation, for example, packeted tablets, capsules, and powders in vials or ampoules. The unit dosage form can also be a capsules, cachet, or tablet itself, or it can be the appropriate number of any of these packaged forms . -24Some of the peptides of the invention were constructed on solid-phase resins designed to produce C-terminal amides either by treatment of the resin with ammonia in methanol or by direct cleavage of an appropriately substituted resin using trifluoroacetic acid, with the required scavengers, giving the amides directly. The latter protocol was used with DuPont RapidAmide® or Nova Biochem Ultrasyn C® resins either in a simple bubbler apparatus (DuPont resin) or automated synthesizer (Nova Biochem resin).

Using a Pharmacia 4170 automated peptide synthesizer and Bioplus® software the peptides were constructed from the C-terminus using Fmoc amino acid pentafluorophenyl or DHBt esters and HOBt catalysis.

Each residue was present in a fivefold excess to ensure rapid and complete acylation. On a 0.095 mmol scale the peptide was isolated following TFA cleavage (94% TFA, 5% anisole, 1% ethanedithiol) from the resin (2 hours, room temperature).

The two isomers were separated by RP-HPLC (250 x 25 mm column, gradient elution, Solvent A 0.1% aqueous TFA, Solvent B 0.1% FFA in MeCN, gradient 20% to 80% B over 20 minutes. The peptides of the invention can be made by the above method. Although any compatible resin may be used or, alternatively, solution phase synthesis may be used.

Such peptides include but are not limited to the short chemotactic peptides, for example, formyl-Met-Leu-Phe-OBz, Met-enkephalin, enkephalinamides, leupeptin (Ac-Leu-Leu-Arginol), Kyotorphin (Tyr-Arg), Morphiceptin (Tyr-Pro-Phe-Pro-NH2), Thymopoetin II (Arg-Lys-Asp-Val—Tyr), Splenopentin (Arg-Lys-Glu-Val-Tyr), Hamburger pentapeptide (Asp-Leu-Asp-Pro-Arg), virus replication inhibiting -25peptide (Z-DPhe-Phe-Gly), DNA binding peptide (Lys-Trp-Lys), Molluscan cardioexcitatory peptide (Phe-Met-Arg-Phe-N^ and all FMRF-amide analogues), TRH (pGlu-His-Pro), Proctolin (Arg-Tyr-Leu-Pro-Thr), Tuftsin, (Thr-Lys-Pro-Arg), Kentsin (Thr-Pro-Arg-Lys), schizophrenia-related peptide (Tnr-Val-Leu) and short polypeptides or 5-residues or less which are fragments of longer peptides.

Preferred compounds are: LTyr-Gly-Gly-a-MePhe-LLeu (isomer 1) and LTyr-Gly-Gly-a-MePhe-LLeu (isomer 2) whose full chemical names are N-[α-methyl-N-[N-(N-Ltyrosylglycyl)glycyl]-L—phenylalanyl]—L-leucine trifluoroacetate (1:1 salt) and N-[α-methyl—Ν-[Ν-(N15 L-tyrosylglycyl)glycyl]-D—phenylalanyl]-L-leucine trifluoroacetate (1:1 salt).

Some of the compounds were evaluated in three tachykinin binding assays: For the NKX receptor - measurement of the binding of [125I]-Bolton Hunter labeled substance P (0.1 nM) to guinea pig cerebral cortex membranes, and for the NK3 receptor - measurement of the binding of [3H]-senktide (2 nM) to guinea pig cerebral cortex membranes. See Lee, C. M., et al, Eur. J. Pharmacol. 130:209 (1986), and Guard, S., et al, Brit. J.

Pharmacol. 99:767 (1990).

For the NK2 receptor - measurement of the binding of [125I]-iodohistidyl neurokinin A (0.1 nM) to hamster urinary bladder membranes. See Buck and Shatzer, Life Sci. 42:2701 (1988).

TABLE III Structure of Results -2610 Ex. X Y Stereochemistry at a-Me-Phe Binding IC50 (nM) ΝΚχ nk2 nk3 7 4-CL nh2 D,L >105 98 >105 8 4-CF3 nh2 D,L >105 188 >105 9 4-Ph nh2 D,L >105 94 >105 10 r^2 nh2 D,L >105 68 >105 nh2 D,L >105 73 >105 NH2 -105 48 >105 13 c nh2 D >10s 1850 >1O5 14 4-CO2-n-Pr nh2 D,L >105 144 >105 18 H nhch2conh2 D,L -105 20 >105 19 (¢^2 nhch2conh2 D,L >105 14 >105 20 Γ2 nhch2conh2 L -105 6 >105 21 2,3—di—OCH3 nh2 D,L 2900 45 >105 -27The data in Table III show that the compounds are selective NK2 receptor ligands. The compounds of Examples 18 and 19 illustrate NK2 receptor antagonism. See Table IV. Therefore, they are 5 expected to be useful in treating disorders mediated by tachykinins, e.g., respiratory disorders, inflammation, gastrointestinal disorders, ophthalmic diseases, allergies, pain, circulatory insufficiency, diseases of the central nervous system, and migraine.

NK2 Functional Data Summary Tissue Agonist Antagonist Cone . Dose\ Ratio pKa Rat Colon Eledoisin Example 18 ι μΜ 3.1 6.5 Example 19 1 μΜ 7.7 6.8 15 Hamster Bladder 1659,874 10 μΜ 10.3 7.0 Urinary NKA Example 18 3 μΜ 30 7.0 Example 19 3 μΜ 29 6.9 20 L659,874 3 μΜ 29 6.9 Hamster Trachea NKA Example 18 1 μΜ 11 6.9 Example 18 3 μΜ 46 7.2 Example 19 3 μΜ 31 6.9 25 Example 19 10 μΜ 472 7.2 L659,874 1 μΜ 11 7.0 L659,874 3 μΜ 41 7.1 a L659,874 is a standard NK2 antagonist. Its chemical name is Glycinamide, N—acetyl—L—leucyl— 0 L-methionyl-L-glutaminyl-L-tryptophylL-phenylalanyl-Ac-LLeu-LMet—LGln—LTrp-LPhe-Gly-NH2 For the hamster trachea, see Maggi, C. A., Patacchini, R., Rovero, P., and Meri, A. Eur. J.

Pharmacol., 1989;166:435-440.

For the hamster urinary bladder, see Mizrahi, J., Dion, S., D'Orleans-Juste, P., Escher, B., Drapeau, G., and Regoli, D., Eur. J. Pharmacol., 1985;118:25-36. -28For the rat colon, see Bailey, S. J. and Jordan, C. C., Br. J. Pharmacol., 1984;82:441-451.

The following examples illustrate the instant invention but are not intended to limit it in any way.

EXAMPLE 1 Tyr-Gly-Gly-OMePhe-Leu The peptide was prepared and separated as described above, using solid-phase methodology.

Isomer 1 (L,D,L or L, L, L) , _LTyr—Gly—Gly— OMePhe-LLeu NMR (D2O) 8 0.89 (6H, br d), 1.41 (3H, s), 1.57 (3H, br m) , 3.18 <2H, 2bq) , 3.12 (2H, d) , 3.92 (4H, m) , 4.25 (2H, m), 6.88 (2H, d), 7.18 (4H, d), 7.46 (3H, m). FAB-MS 57C (M+H)+, 592 (M+Na)+, 608 (M+K)+.

Isomer 2 (L,L,L or L,D,L) LTyr-Gly-Gly-OCMePhe-LLeu NMR (D2O) δ 0.88 (6H, dd), 1.43 (3H, s), 1.57 (3H, br m), 3.20 (4H, m), 3.91 (4H, m), 4.23 (2H, m), 6.88 (2H, d), 7.16 (4H, d), 7.33 (3H, m). FAB-MS 570 (M+H)+, 592 (M+Na)+, 608 (M+K)+.

EXAMPLE 2 N—(t-Butyloxycarbonyl)-DL—a-methylphenylalanine (1) DL-a-Methylphenylalanine (5.0 g, 28 mmol) was dissolved in warm 10% Na2CQ3 solution (60 mL) and then cooled to 0°C. t-Butyloxycarbonyl anhydride (6.39 g, 29.3 mmol) in dioxan (50 mL) was added dropwise and the mixture stirred at 0°C for 1 hour.

The mixture was then allowed to warm to room temperature and stirred for a further 24 hours. The solvents were then distilled off in vacuo and the residue taken up in H20. This was then washed with CH2C12 (3x), acidified with citric acid, and -29extracted with CH2C12 (3x). The organic extracts were combined, dried (MgSO4) , and the solvent distilled off in vacuo to give 1 as a white solid (6.2 g, 25.1 mmol, 90%): ΧΗ NMR (DMSO-dg) δ 1.18 (3H, s), 1.41 (9H, s) , 2.91 (IH, d, J 13 Hz), 3.31 (IH, d, J 13 Hz), 6.71 (IH, bs), 7.09 (2H, d), 7.25 (3H m) , 12.50 (IH, bs) .

EXAMPLE 3 N-(t-Butyloxycarbonyl)-DL—a—methylphenylalaninamide 111 (6.7 g, 22 mmol) was dissolved in CH2C12 (80 mL) and 1,3-dicyclohexylcarbodiimide (4.9 g, mmol) followed by 1-hydroxybenzotriazole monohydrate (3.9 g, 28.8 mmol) added and the mixture stirred for 0.5 hour at room temperature. DMF (15 mL) was then added, the solution cooled to -10°C and a slow stream of NH3 (g) bubbled through. The mixture thickened almost immediately so a further amount of DMF (50 mL) was added. After 0.5 hour, the addition of NH3 (g) was stopped and the white precipitate removed by filtration and washed with EtOAc. The washings were combined with the filtrate and the solvent distilled off in vacuo. The residue was partitioned between H2O (500 mL) and CH2C12. The aqueous layer was washed with CHC12 (3x), the organic extracts combined, washed with a saturated NaHCO3 solution, dried (MgSO4) , and the solvent distilled off in vacuo. The white solid obtained was further purified by flash column chromatography on silica, eluting with a mixture of CHCl3:MeOH (95:5) to give 2 as a white solid (4.4 g, 15.8 mmol, 66%): 1H NMR (DMSO-d6) δ 1.33 (3H, s), 1.41 (9H, s), 3.31 (2H, bs), 6.25 (IH, bs), 7.15 (6H, m), 7.41 (IH, bs). -30EXAMPLE 4 DL-q-Methylphenylalaninamide (3) (4.4 g, 15 mmol) was stirred in TFA (15 mL) at room temperature for 15 minutes. The TFA was distilled off in vacuo and the residue taken up in EtOAc. The organic solution was carefully washed with saturated NaHCO3 solution, dried (MgSO4) , and the solvent removed in vacuo to give 3 as a white solid (1.9 g, 12.1 mmol, 70%) ΧΗ NMR (MeOH-d4) δ 1.40 (3H, s), 2.77 (1H, d, J 13 Hz), 3.21 (1H, d, J 13 Hz), 7.30 (5H, m).

EXAMPLE 5 N—(9H-Fluoren-9-ylmethoxv)carbonyl)-L-tryptophyl—DL15 g-methylphenylalaninamide (4) (1.8 g, 10.1 mmol) and N-((9H-fluoren-9ylmethoxy)carbonyl]—L-tryptophan pentafluorophenyl ester (6.0 g, 10.1 mmol) were stirred together in DMF (50 mL) at room temperature for 18 hours. The solution was concentrated in vacuo and H2O (400 mL) added. The aqueous suspension was extracted with EtOAc (3x) and the organic extracts combined, dried (MgSO4) and the solvent removed in vacuo to give crude 4.

EXAMPLE 6 L-Tryptophyl-DL-g—methylphenylalaninamide (5) (3.65 g, 6.2 mmol) was dissolved in a 20% solution of piperidine in DMF (20 mL) and stirred at room temperature for 20 minutes. The mixture was then concentrated in vacuo and H2O (400 mL) added; the white precipitate formed was then removed by filtration. Citric acid was added to the filtrate to pH 3 and washed with EtOAc (3x). The aqueous layer was made alkaline (pH 9) with solid Na2CO3 and -31extracted with EtOAc (3x). The organic extracts were combined, dried (MgSO4) and the solvent removed in vacuo to give 5 as a pale yellow solid (2.2 g). Purification by flash column chromatography on silica, eluting with CH2Cl2:MeOH (10:1) gave the product 5 as a white foam (1.8 g, 4.93 mmol, 80%): XH NMR (DMSO—dg) δ 1.45 (0.5 x 3H, s), 1.51 (0.5 X 3H, s), 1.81 (2H, bs), 2.59 (0.5 x H, dd, J 10.14 Hz), 2.76 (0.5 x H, dd, 9, 14 Hz) , 3.15 (2H m), 3.45 (2H, m), 6.95- 7.60 (12H, m), 8.16 (0.5 x H s), 8.28 (0.5 x H, s), 10.85 (IH, s).

EXAMPLE 7 N—[[(4-Chlorophenyl)methoxy]carbonyl]-L-tryptophyl15 g-methyl-DL-phenvlalaninamide (6) Pyridine (0.11 mL, 1.4 mmol) was added dropwise to a stirred solution of 4-chlorobenzyl alcohol (0.20 g, 1.4 mmol) and triphosgene (0.15 g, 0.51 mmol) in CH2C12 (5 mL) at 0°C and stirred for minutes. The solvent was removed in vacuo and the resulting oil triturated with Et2O and filtered. The filtrate was added to a solution of 5 (0.26 g, 0.71 mmol) and triethylamine (0.10 mL, 0.71 mmol, in THF (25 mL) and stirred overnight. The solvent was removed in vacuo to give a white solid which was partitioned between EtOAc and 10% citric acid solution. The organic layer was washed with saturated NaHCO3 solution and H2O and dried (MgSO4) . Further purification by flash column chromatography on silica eluting with a mixture of CH2Cl2:MeOH (95:5) gave 6 as a white foam (0.23 g, 0.43 mmol, 61%); mp 89-101.3°C; MS (FAB) m/e 533 [M+H]: XH NMR (DMSO—dg) δ 1.38 (0.5 x 3H, s, a-CH3) , 1.4 0 (0.5 x 3H, s, a-CH3) , 2.80-3.40 (4H, m, CH2-indole, CH2Ph), 4.10-4.32 (IH, m, Trp a-H), 4.80-4.99 (2H, m, -32CH2O), 6.90-7.78 (18H, m, Ar, CONH2, CONH, OCONH), .70 (IH, s, indole NH) ; Anal. (C29H29N4O4C ♦ 0.3 H2O) C, Η, N.

EXAMPLE 8 N-[[[4-(Trifluoromethyl)phenyl]methoxy]carbonyl]L-tryptophyl-a-methyl-DL—phenylalaninamide (7) Prepared by the same method as 6. White foam (0.092 g, 62%); mp 91-112°C; MS (FAB) m/e 567 [M+H]; 'H NMR (DMSO—dg) δ 1.38 (0.5 χ 3H, s, a-CH3) , 1.41 (0.5 χ 3H, s, a-CH3), 2.70, 3.46 (4H, m, CH2-indole, CH2Ph), 4.12-4.36 (IH, m, Trp a-H), 4.90-5.16 (2H, m, CH2O), 6.90-7.88 (18H, m, Ar, CONH2 CONH, OCONH), 10.84 (IH, s, indole NH); Anal. (C30H29N4O4F3 · 0.75 H2O) C, Η, N.

EXAMPLE 9 Ν-ί(fl,1'-Biphenyl]-4-ylmethoxv)carbonyl]-Ltryptophyl-a-methyl—DL—phenylalaninamide (8) Prepared by the same method as 6. White foam (0.050 g, 32%); mp 92-102°C; MS (FAB) m/e 576 [M+H]; 1H NMR (DMSO—d6) 8 1.38 (0.5 χ 3H, s, a-CH3) , 1.39 (0.5 χ 3H, s, a-CH3) , 2.72-3.40 (4H, m, CH2-indole, CH2Ph), 4.12-4.35 (IH, m, Trp a-H), 4.90-5.08 (2H, m, CH2O) , 6.92-7.85 (23H, m, Ar, CONH2, CONH, OCONH), .90 (IH, s, indole NH); Anal. (C35H34N4O4·0.75 H2O) C, Η, N.

EXAMPLE 10 N—[(9-Anthracenvlmethoxv)carbonyl1-L-tryptophyl-amethyl-DL-phenylalaninamide (9) Prepared by the same method as 6. Yellow foam (0.10 g, 23%); mp 113-130°C; MS (FAB) m/e 599 [M+H]; XH NMR (DMSO—d6) 1.36 (3H, s, a-CH3) , 2.25-3.20 (2H, m, CH2-indole), 3.25-3.40 (2H, m, CH2Ph), 4.20-4.38 -33(1H, m, Trp α-H), 5.95 (0.5 χ 2H, AB, J 12 Hz, CH2Ar), 6.01 (0.5 χ 2H, AB, J 12 Hz, CH2Ar) , 6.857.20 (17H, m, Ar, CONH2, OCONH), 7.77 (0.5 χ H, s, CONH), 7.83 (0.5 χ H, s, CONH), 8.10 (IH, s, Ar), 8.13 (IH, s, Ar), 8.28 (IH, s, Ar), 8.31 (IH, s, Ar), 8.67 (IH, s, Ar), 10.80 (IH, s, indole NH); Anal. (C37H34N4O4 · 0.7 H2O) C, H, N.

EXAMPLE 11 N-[(1-Naphthalenylmethoxy)carbonyl]-L-tryptophyl-amethyl-DL—phenylalaninamide (10) Prepared by the same method as 6. White foam (0.13 g, 34%); rrp 102-113°C; MS (FAB) m/e 549 [M+H]; ΧΗ NMR (DMSO-dg) δ 1.40 (3H, s) , 2.87 (IH, m), 3.05lS 3.45 (3H, m), 4.15-4.40 (IH, m), 5.30-5.50 (2H, m), 6.90-8.00 (21H, m) , 10.70 (IH, s); Anal. (C33H32N4O40.4 H2O) C, Η, N.

EXAMPLE 12 Ν-Γ(1-Naphthalenylmethoxy)carbonyl]-L-tryptophylq-methyl-L-phenylalaninanride (11) Prepared by the same method as 6, -a-methylL—phenylalanine was obtained by the method of Turk, et al (Turk, J., Panse, G. T., Marshall G. R., J.

Org♦ Chem. 40:953 (1975)).

EXAMPLE 13 Ν-Γ(1-Naphthalenylmethoxy)carbonyl)-L-tryptophyl-ftmethyl—D—phenylalaninamide (12) Prepared by the same method as 6. D-a-methylphenylalanine was obtained by the method of Turk, et al. -34EXAMPLE 14 N-[[Γ4-(Propoxycarbonyl)phenyl]methoxy]carbonyl]-Ltryptophyl-a-methyl-DL-phenvlalaninamide (13) Propyl 4-(hydroxymethyl)benzoate (0.10 g, 0.5 mmol), 4-nitrophenyl chloroformate (0.10 g, 0.5 mmol) and pyridine (0.04 mL, 0.5 mmol) were stirred for 18 hours in CH2C12 (8 mL) . The solvent was removed in vacuo and the white residue triturated with EtOAc and filtered. The filtrate was added to a solution of 5 (0.10 g, 0.27 mmol) and 1,1,3,3-tetramethylguanidine (0.10 mL, 0.81 mmol) in DMSO (2 mL) and stirred for 3.5 hours. The reaction mixture was poured onto H20 and extracted with EtOAc. The organic layer was washed with 10% citric acid solution, saturated NaHCO3 solution, H20, and dried (MgSO4) . Further purification by flash column chromatography on silica, eluting with CH2C12 (95:5) gave 13 as a white foam (0.02 g, 13%); mp 78-105°C; MS (FAB) m/e 586 [M+H]; XH NMR (DMSO-d6) δ 0.97 (3H, t, J 7 Hz, CH3CH2), 1.38 (0.5 x 3H, s, a-CH3) , 1.40 (0.5 x 3H, s, a-CH2), 1.76 (2H, m, CH2CH2), 2.80-3.45 (4H, m, CH2-indole, CH2Ph) , 4.12-4.35 (3H, m, Trp a-H, CH2CH2O) , 4.94-5.15 (2H, m, CH2OCO) , 6.95-7.95 (18H, m, Ar, CONH2, CONH, OCONH, 10.82 (IH, s, indole NH) ; Anal. (C33H36N4O6 · 0.75 H2O) C, Η, N.

EXAMPLE 15 Boc(a-Me)-DL-PheODhbt (14) 1,3-Dicyclohexylcarbodiimide (4.95 g, 24 mmol) was added to a solution of 1 (6.70 g, 24 mmol) in THF (100 mL) at -15°C and allowed to stir for 5 minutes. 3-Hydroxy-l,2,3-benzotriazine—4-(3H)-one (3.91 g, mmol) and an additional volume of THF (20 mL) were added and the mixture stirred at —10°C for 1 hour and at 0°C for 4 hours. After leaving overnight at 5°C a -35white solid precipitated. This was filtered and dried to give 14 (9.1 g, 89%): 1H NMR (DMSO-dg) δ 1.44 (3H, s, a-CH3), 1.49 (9H, s, t-Bu), 3.06 (1H, d, J 13 Hz, CH'HPh), 3.61 (1H, d, J 13 Hz, CH'HPh), 7.15-7.38 (5H, m, Ph), 7.64 (1H, s, OCONH), 8.02 (1H, , J 7.5 Hz, Ar), 8.19 (1H, t, J 7.5 Hz, Ar), 8.33 (2H, d, J 8 Hz, Ar).

EXAMPLE 16 Boc-(«-Me)-DLPheGlyNH, (15) (4.24 g, 10 mmol), glycinamide hydrochloride (1.11 g, 11 mmol), and triethylamine (1.39 mL, mmol) in EtOAc (100 mL) were stirred overnight at room temperature. The white solid which precipitated was removed by filtration and washed with EtOAc. The filtrate was washed with 10% citric acid solution, saturated NaHCO3 solution, H2O, and dried (MgSO4) . Removal of the solvent in vacuo gave 15 as a white solid (2.05 g, 61%): 1H NMR (DMSO-d6) 6 1.16 (3H, s, a-CH3) , 1.43 (9H, s, t-Bu), 2.95 (1H, d, J 13 Hz, CH2Ph), 3.26 (1H, d, J 15 Hz, CH2Ph), 3.46 (1H, br d, J 16.5 Hz, NHCH2CO) , 3.69 (1H, dd, J 16.5 Hz, 6 Hz, NHCH2CO) , 7.02-7.34 (8H, m, Ar, CONH2, OCONH), 8.17 (1H, bs, NHCO).

EXAMPLE 17 (g-Me)-DLPheGlvNH2 (16) (2.0 g, 5.7 mmol) was stirred at 0°C in TFA (2 mL) for 15 minutes. The solvent was removed in vacuo and the residue triturated with ET2O. The white solid formed was filtered and dried to give 16 (1.97 g, 95%): 1H NMR (DMSO-d6) δ 1.52 (3H, s, a-CH3), 3.06 (1H, d, J 14 Hz, CH2Ph), 3.20 (1H, d, J 14 Hz, CH2Ph), 3.74 (2H, d, J 5.5 Hz, NHCH2CO) , -367.00—7.40 (7H, m, Ph2CONH2) , 8.09 (3H, bs, NH3+), 8.62 (1H, J 5.6 Hz, CONH).

EXAMPLE 18 N-[(Phenylmethoxy)carbonyl]—L-tryptophyl-a-methyl-DLphenyl alanvlqlvcinanu.de (14) Hydrobenzotriazolyl tetramethyluronium hexachlorophosphate (HBTU) (0.19 g, 0.5 mmol) was added to a solution of Z-(L)-Trp (0.15 g, 0.5 mmol), 16 (0.17 g, 0.5 mmol), and N,N-diisopropylethylamine (0.26 mL, 1.5 mmol) in DMF (3 mL) and stirred at room temperature for 45 minutes. Water was added and the mixture extracted with EtOAc. The organic layer was washed with 10% citric acid solution, saturated NaHCO3 solution, H2O, and dried (MgSO4) . Further purification by flash column chromatography on silica, eluting with CH2Cl2:MeOH (90:10) gave 17 as a white foam (0.17 g, 61%); mp 92-110°C; MS (FAB) m/e 556 [M+H]; ΧΗ NMR (DMSO-d6) 5 1.19 (0.5 x 3H, S, a-CH3) , 1.24 (0.5 x 3H, s, a-CH3) , 2.80-3.30 (4H, m, CH2-indole, CH2Ph), 3.40-3.78 (2H, m, CH2CONH2) , 4.25-4.48 (1H, m, Trp-a-H), 4.84-5.06 (2H, m, CH2OCO), 6.88-7.45 (16H, m, Ar, OCONH, CONH2) , 7.55-7.75 (2H, m, Ar), 7.80 (0.5 x H, t, CONHCH2) , 7.88 (0.5 x H, t, CONHCH2) , 8.35 (0.5 x H, s, NHC(CH3), 8.41 (0.5 x H, s, NHC(CH3), 10.86 (1H, s, indole NH) ; Anal. (C31H33N5O35· 0.5H2O) C, Η, N.

EXAMPLE 19 N-F(1-Naphthalenylmethoxy)carbonyl]-L-tryptophyl-amethyl-DL-phenylalanylqlycinamide (15) Diisopropylethylamine (52 μΐ, 0.3 mmol) was added to a stirred solution of N—[(1-naphthylmethoxy)carbonyl]tryptophan (0.117 g, 0.3 mmol) and HBTU (0.114 g, 0.3 mmol) in DMF (5 mL) at room -37temperature. The mixture was stirred for 10 minutes, then 16 (0.105 g, 0.3 mmol) in DMF (5 mL) was added, followed by diisopropyl ethylamine (105 μL, 0.6 mmol) and the reaction mixture stirred for a further 18 hours. The mixture was poured into H20 (100 mL) and extracted with EtOAc (2 x 50 mL). The organic extracts were combined, washed with 10% citric acid solution (1 x 100 mL), saturated NaHCO3 solution (1 x 100 mL), H2O (2 x 100 mL) , and dried (MgSO4) .

The solvent was removed in vacuo and the residue purified by column chromatography on silica, eluting first with mixtures of EtOAc in hexane (30% to 70%) followed by CH2Cl2:MeOH (95:5) to give 18 as a white solid (0.148 g, 81%); MS (FAB) m/e 606 [M+H]: τΗ NMR (DMSO-dg) 0 1.21 (0.5 x 3H, s, a-CH3) , 1.26 (0.5 x 3H, s, a-CH3) , 2.85-3.78 (6H, m, CH2-indole, CH2Ph, Ch2CONH2) , 4.38 (1H, m, Trp a-H) , 5.44 (2H, m, CH2 naphthyl), 6.92-7.20 (10H, m, Ar/NH), 7.33 (0.5 x H, s, Ar/NH), 7.35 (0.5 x H, s, Ar/NH), 7.52-7.70 (6H, m, Ar/NH), 7.75-8.00 (4H, m, Ar/NH), 8.33 (0.5 x H, s, NH), 8.35 (0.5 x H, s, NH), 10.72 (1H, s, NH indole); Anal. (C35H35N5O5 · 0.5H2O) C, Η, N.

EXAMPLE 20 N—[(1-Naphthalenylmethoxy)carbonyl]-L-tryptophyl-qmethyl-L-phenvlalanylqlvcinamide (16) Prepared by the same method as Compound 18 as in Example 19 in methylphenylalanine and obtained by the method of Tuttle, et al.

EXAMPLE 21 N-[[(2,3-Dimethoxyphenyl)methoxy]carbonyl]L-tryptophyl-a-methyl-DL-phenylalaninamide (17) Prepared by the same method as Compound 6 in 35 Example 7, white foam: mp 87-102°C; MS (FAB) m/e 559 -38[M+H] ; ΧΗ NMR (DMSO-dg) δ 1.39 (3H, S,CC-CH3) , 2.88 (IH, t) and 3.11-3.39 (3H, m, CH2-indole, CH2Ph), 3.63 (3H, d, OCH3), 3.78 (3H, s, OCH3) , 4.21 (IH, m, Trpa-H), 4.93 (2H, s, CH2O) , 6.79-7.78 (17H, m, Ar, CONH2, CONH, OCONH), 10.76 (IH, s, indole NH), and <C31H34N4°6·0·5 H2°) C' H' NIE 921321

Claims (23)

1. A compound of formula R 1 —NH—C—COR 3 I l 4 R or a pharmaceutically acceptable salt thereof wherein: r1 is an N-terminal blocking group, from 0 to 4 amino acid residues or hydrogen; R is a sidechain of a genetically coded amino acid except glycine; R is a C-terminal blocking group from 0 to 4 amino acid residues, -OH, or OR n wherein R n is straight or branched alkyl or cycloalkyl of from 1 to 6 carbon atoms; R is a sidechain of a genetically coded ammo acid, except glycine, or -hc=ch 2 , -CsCH, -ch 2 -ch=ch 2 , -ch 2 c=ch, -CH 2 Ar, -ch 2 or, -CH 2 OAr, -( CH 2) n C0 2Rf oe - (CH 2 ) n NR 3 R.6 wherein n is an integer of from 0 to 3, R is hydrogen or lower alkyl, Ar is a mono- or polycyclic unsubstituted or substituted carbo- or heterocyclic aromatic or hydroaromatic moiety; -40R 4 and R 2 cannot be hydrogen; R 1 and R 3 together cannot be more than 4 35 amino acid residues.

2. A compound according to Claim 1 wherein: R 1 is H, Boc, Fmoc, 5 Z, 1- Adoc, 2- Adoc, IVA, or NV A; 10 R 2 is CHg- or HOOC-CH 2 - ; and R 3 is “NH 2 , -OCH^, or -OCH 2 Ph.

3. A compound according to Claim 1 selected from MeLys-Trp-Asp-Asn-Gln, Lys—MeTrp-Asp—Asn—Gin, Lys-Trp-MeAsp-Asn—Gin, 5 Lys-Trp-Asp-MeAsn-Gln, Lys-Trp-Asp-Asn—MeGln, MeVal-Gly-His-Leu-Met-NH 2 , Val-Gly—MeHis-Leu—Met-NH 2 , Val—Gly—His—MeLeu—Met-NH 2 , 10 Val-Gly-His-Leu-MeMet—NH 2 , MeGlp-His-Trp-Ser-Tyr, Glp-MeHi s-Trp-Ser-Tyr, Glp-His-MeTrp-Ser-Tyr, Glp-His-Trp-MeSer-Tyr, 15 Glp-His-Trp-Ser-MeTyr, Gly-MeLeu-Arg-Pro—Gly—NH 2 , —41— Gly-Leu-MeArg-Pro-Gly—NH 2 , Gly-Leu-Arg-MePro-Gly—NH 2 , MeTyr-Pro-Ser-Lys-Pro, 20 Tyr-MePro-Ser-Lys-Pro, Tyr-Pro-MeSer-Lys-Pro, Tyr-Pro-Ser-MeLys-Pro, Tyr-Pro-Ser-Lys-MePro, MeThr-Arg-Gln-Arg-Tyr-NH 2f 25 Thr-MeArg-Gln-Arg-Tyr-NH 2 , Thr-Arg-MeGln—Arg-Tyr-NH 2 , Thr—Arg-Gln-MeArg-Tyr-NH 2 , Thr-Arg-Gln-Arg-MeTyr-NH 2 , Gly—MeTrp—Thr—Leu—Asn, 30 Gly—Trp—MeThr—Leu-Asn, Gly-Trp-Thr-MeLeu-Asn, Gly-Trp-Thr-Leu-MeAsn, MeLeu-Tyr-Gly-Leu-Ala-NH 2 , Leu—MeTyr—Gly—Leu—Ala—NH 2 , 35 Leu—Tyr—Gly-MeLeu-Ala—NH 2 , Leu-Tyr-Gly-Leu—Aib-NH 2 , MePhe-Phe-Trp-Lys-Thr, Phe-MePhe-Trp-Lys-Thr, Phe-Phe-MeTrp-Lys-Thr, 40 Phe-Phe-Trp-MeLys-Thr, Phe-Phe-Trp-Lys-MeThr, MePhe-Phe—Gly-Leu-Met—NH 2 , Phe-MePhe-Gly-Leu-Met-NH 2 , Phe—Phe—Gly-MeLeu-Met—NH 2 ,

4. 5 Phe-Phe-Gly-Leu-MeMet-NH 2 . and

A compound named N—[ 4. -425. A compound named N-[α-methyl-N-[N-(N-Ltyrosylglycyl)glycyl]-D-phenylalanyl]-L-leucine trifluoroacetate (1:1 salt).

6. A compound named N-[[(4-chlorophenyl)methoxy]carbonyl]-L-tryptophyl—a—methyl-DLphenylalaninamide.

7. A compound named N-[[[4-(trifluoromethyl)phenyl] methoxy] carbonyl] -L-tryptophyl—OC-methylDL—phenylalaninamide.

8. A compound named N-[([1,1'-biphenyl]-4-ylmethoxy) carbonyl] -L-t rypt ophvl-Ct-methyl-DLphenylalaninamide.

9. A compound named N-[(9-anthracenylmethoxy)— carbonyl]-L-tryptophyΙ-α-methy1-DLphenylalaninamide.

10. A compound named N—[(1—naphthalenylmethoxy)— carbonyl]-L-tryptophyl—a-methyl—DLphenylalaninamide.

11. A compound named N-[(1—naphthalenylmethoxy)carbonyl]-L-tryptophy1-a—methy1—Lpheny1a1aninamide.

12. A compound named N—[(1—naphthalenylmethoxy) — carbonyl]-L-tryptophyl-a-methyl-Dphenylalaninamide.

13. A compound named N-[[[4-(propoxycarbonyl)phenyl]methoxy]carbonyl]—L-tryptophy1-a-methy1DL-phenylalaninamide.

-4314. A compound named N-[(phenylmethoxy)carbonyl]-Ltryptophyl-a-methyl-DL-phenylalanylglveinamide.

15. A compound named N-[(1—naphthalenylmethoxy) carbonyl]-L-tryptophyl—q-methyl-DL-phenylalany1glycinamide.

16. A compound named N-[(1-naphthalenylmethoxy)carbonyl]-L-tryptophyl-a-methyl-Lphenylalanylglycinamide.

17. A compound named N-[[(2,3—dimethoxyphenyl)methoxy]carbonyl]-L-tryptophyΙ-α-methyl-DLphenylalaninamide.

18. A pharmaceutical composition comprising an amount of a compound according to Claim 1 effective to treat pain in a mammal suffering therefrom, and a pharmaceutically acceptable 5. Carrier.

19. A method of treating pain in a mammal comprising administering an effective pain treating amount of a compound according to Claim 1.

-4420. A compound as claimed in Claim 1, substantially as hereinbefore described and exemplified.

21. A pharmaceutical composition according to Claim 18, substantially as hereinbefore described.

22. A process for preparing a compound as claimed in Claim 1, substantially as hereinbefore described and exemplified.

23. A compound as claimed in Claim 1, whenever prepared by a process claimed in Claim 22.