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WO1994009031A1 - Analogues of cholecystokinin (30-33) containing an alpha-substituted aminoacid - Google Patents

Analogues of cholecystokinin (30-33) containing an alpha-substituted aminoacid Download PDF

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Publication number
WO1994009031A1
WO1994009031A1 PCT/US1993/009809 US9309809W WO9409031A1 WO 1994009031 A1 WO1994009031 A1 WO 1994009031A1 US 9309809 W US9309809 W US 9309809W WO 9409031 A1 WO9409031 A1 WO 9409031A1
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Prior art keywords
methyl
gly
tryptophyl
carbonyl
leu
Prior art date
Application number
PCT/US1993/009809
Other languages
French (fr)
Inventor
David Christopher Horwell
William Howson
John Hugues
Reginald Stewart Richardson
Original Assignee
Warner-Lambert Company
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Warner-Lambert Company filed Critical Warner-Lambert Company
Priority to AU53596/94A priority Critical patent/AU5359694A/en
Publication of WO1994009031A1 publication Critical patent/WO1994009031A1/en

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06139—Dipeptides with the first amino acid being heterocyclic
    • C07K5/06156—Dipeptides with the first amino acid being heterocyclic and Trp-amino acid; Derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/02—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link
    • C07K5/0202—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link containing the structure -NH-X-X-C(=0)-, X being an optionally substituted carbon atom or a heteroatom, e.g. beta-amino acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/08—Tripeptides
    • C07K5/0821—Tripeptides with the first amino acid being heterocyclic, e.g. His, Pro, Trp
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • 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
  • modified peptides offer significant opportunities.
  • the ⁇ , ⁇ -disubstituted amino acids are non-genetically coded synthetic analogues of natural mammalian ⁇ -amino acids and are incorporated at least once into the
  • the invention relates to novel compounds of formula
  • 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.
  • the invention also relates to a pharmaceutical composition for reducing gastric acid secretion
  • 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 method for
  • the invention further relates to a method for 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 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.
  • 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.
  • the invention also relates to a method for
  • the invention also relates to a method for
  • 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.
  • 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,
  • the invention further provides processes for the preparation of compounds of Formula I.
  • the 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.
  • N-terminal protecting 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,
  • C-terminal protecting group 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.
  • side chains and protecting groups are those known in the art. Any of those could be used.
  • R 1 is an N-terminal blocking group or from 0 to
  • R is a sidechain of a genetically coded ammo
  • R is a C-terminal blocking group or from 0 to
  • R n is straight or branched alkyl or cycloalkyl of 1 to 6 carbon atoms
  • R 4 is a sidechain of a genetically coded ammo
  • 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
  • Preferred compounds of the invention are those of Formula I selected from:
  • More preferred compounds of the invention are those of Formula I selected from:
  • 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 recrystallization and reconversion to the nonsalt form.
  • the compounds of the present invention may be formed by coupling individual substituted ⁇ -amino acids by methods well known in the art. (See, for example, standard synthetic methods discussed in the
  • inert, pharmaceutically acceptable carriers can be either solid or liquid.
  • Solid form preparations include powders, tablets, dispersible granules, capsules, cachets, and
  • 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.
  • the carrier is a finely divided solid which is in a mixture with the finely divided active component.
  • the active component is mixed with the carrier having the necessary binding
  • 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.
  • 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 which is thus in association with it.
  • encapsulating material as a carrier providing a capsule in which the active component (with or without other carriers) is surrounded by a carrier which is thus in association with it.
  • cachets are included. Tablets, powders, cachets, and capsules can be used as solid dosage forms suitable for oral
  • Liquid form preparations include solutions, suspensions, and emulsions. Sterile water or
  • 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
  • the pharmaceutical preparation is in unit dosage form.
  • the preparation is in unit dosage form.
  • the preparation is divided into unit doses containing appropriate
  • the unit dosage form can be a packaged preparation, the package
  • 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.
  • RapidAmide ® or Nova Bi.ochem Ultrasyn C ® resins either in a simple bubbler apparatus (DuPont resin) or automated synthesizer (Nova Biochem resin).
  • Such peptides include but are not limited to the short chemotactic peptides, for example,
  • Preferred compounds are:
  • LTyr-Gly-Gly- ⁇ -MePhe-LLeu (isomer 2) whose full chemical names are N-[ ⁇ -methyl-N-[N-(N-L-tyrosylglycyl)glycyl]-L-phenylalanyl]-L-leucine trifluoroacetate (1:1 salt) and N-[ ⁇ -methyl-N-[N-(N-L-tyrosylglycyl)glycyl]-D-phenylalanyl]-L-leucine trifluoroacetate (1:1 salt).
  • NK 1 receptor - measurement of the binding of [ 125 I]-Bolton Hunter labeled substance P (0.1 nM) to guinea pig cerebral cortex membranes and for the NK 3 receptor - measurement of the binding of [ 3 H]-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
  • Examples 18 and 19 illustrate NK 2 receptor antagonism. See Table IV below. Therefore, they are expected to be useful in treating disorders mediated by tachykinins, eg, respiratory disorders, inflammation, gastrointestinal disorders, ophthalmic diseases, allergies, pain, circulatory insufficiency, diseases of the central nervous system, and migraine. TABLE IV. NK 2 Functional Data Summary
  • L659,874 3 ⁇ M 41 7.1 a L659,874 is a standard NK 2 antagonist. Its structure is
  • the peptide was prepared and separated as
  • 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 Na 2 CO 3 and extracted with
  • 1,3-Dicyclohexylcarbodiimide (4.95 g, 24 mmol) was added to a solution of 1 (6.70 g, 24 mmol) in THF
  • hexachlorophosphate (0.19 g, 0.5 mmol) was added to a solution of Z-(L)-Trp (0.15 g, 0.5 mmol), 16
  • Trp- ⁇ -H 4.84-5.06 (2H, m, CH 2 OCO) , 6.88-7.45 (16H, m, Ar, OCONH, CONH 2 ) , 7.55-7.75 (2H, m, Ar) , 7.80
  • Step 4 To a solution of the acid (Step 4) (216 mg) in DMF (4 mL) was added the HCl-GlyNH 2 (41 mg) followed by BOP reagent (164 mg) and DIPEA (144 mg). The reaction mixture was stirred overnight at room temperature and then evaporated to dryness. The residue was taken up in EtOAc and the organic layer washed with aqueous 0.1 M HCl solution. The organic layer was then
  • Acid (14) was prepared as a crude sample without purification and was a white amorphous solid (90 mg, 93%); IR (film) 3340, 1721, 1715, and 1667 cm -1 .
  • Example 7 white solid (0.100 g, 58%); MS (FAB) m/e 603 [M+H]; 1 H NMR (DMSO-d 6 ) ⁇ 1.39 (3H, s, ⁇ -CH 3 ), 2.89-3.38 (4H, m, CH 2 indole, CH 2 Ph), 4.02 (1H, m, Trp ⁇ H), 5.07 (2H, m, Ar CH 2 O 2 CNH) , 6.97-7.85 (23H, m, Ar, NH, NH 2 , OCONH), 10.84 (1H, s, indole NH); Anal.
  • Example 7 yellow foam (0.37 g, 68%); MS (FAB) m/e 544 [M+H]; 1 H NMR (DMSO-d 6 ) ⁇ 1.39 (0.5 ⁇ 3H, S, ⁇ -CH 3 ), 1.42 (0.5 ⁇ 3H, s, ⁇ -CH 3 ), 3.16-3.40 (4H, m,
  • 1,3-Dicyclohexyxcarbondiimide (4.95 g, 24 mmol) was added to a solution of ⁇ -methyl-L-phenylalanine (obtained by the method of Turk, et al) (6.70 g,
  • N-BOC- ⁇ -Me-(L)Phe(H)-DCHA (4.6 g, 10 mmol) was taken up in CH 2 Cl 2 and washed with 1N HCl, H 2 O, dried (MgSO 4 ), and the solvent was removed in vacuo to give the acid as a solid.
  • the solid was taken up in DMF (50 mL) and stirred with HBTU (3.80 g, 10 mmol) and diisopropylethyl amine (2.58 g, 20 mmol) at room temperature for 30 minutes.

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Abstract

The compounds of the invention are α-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, cancers, asthma, bladder dysfunction, psychosis, and arthritis and/or inflammatory pain. Pharmaceutical compositions, novel intermediates, and processes are also included.

Description

ANALOGUES OF CHOLECYSTOKININ (30-33) CONTAINING AN ALPHA-SUBSTITUTED AMINOACID.
BACKGROUND OF THE INVENTION
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 α,α-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.
Some examples of therapeutic applications of these modified peptides (peptoids) are given in Table I below. TABLE I
Endogenous Peptide Peptoid
Type* Therapy
Angiotensin ANT Hypertension
Heart Failure
Atrial Natriuretic Factor AG Heart Failure
Thyrotropin Releasing AG Stroke
Factor Cognition
Spasticity
Depression
Neuropeptide-Y ANT Hypertension
Depression
Obesity
Glucagon ANT Diabetes
Insulin AG Diabetes
Gastrin ANT Gastric Ulcer
Cancer
Bombesin ANT Cancer
Tachykinins ANT Antipsychotic
SP, NKA, NKB 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 is 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 Increased Sexual Factor Receptivity
Corticotrophin Releasing Anxiety
Factor Enhanced Vigilance
Thyrotropin Releasing Factor Increased Activity
Enhanced Awareness
SP or NKA A dopamine
behavioral
syndrome, i .e
increased
locomotion, rearing
SP Antidipsogenic
activity
SUMMARY OF THE INVENTION
The invention relates to novel compounds of formula
Figure imgf000005_0001
and the pharmaceutically acceptable salts thereof wherein R1, R2, R3, and R4 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.
The 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
compound 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 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 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
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 dysfunction. 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.
The 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.
Figure imgf000010_0001
Figure imgf000011_0001
N-Terminal
Protecting
Abbreviation Group
H Hydrogen
Tert-butyloxy¬
BOC carbonyl
CBZ Benzyloxy(or Z) carbonyl
IBU Isobutyryl
IVA Isovaleryl
NVA n-Valeryl
1-Adamantyloxy¬
1-AdoC carbonyl N-Terminal
Protecting
Abbreviation Group
2-Adamantyloxy¬
2-Adoc carbonyl
9-Fluorenyl¬
Fmoc methoxycarbonyl
C-Terminal
Substituent/
Side Chain
Protecting
Abbreviation Group
-NH2 Amide
-OCH3 Methyl ester
Benzyl
-OCH2Ph ester/ether
Tert-butyl
-OC(CH3)3 ester/ether
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
Figure imgf000013_0001
and the pharmaceutically acceptable salts thereof, wherein
R1 is an N-terminal blocking group or from 0 to
4 amino acid residues or hydrogen;
2
R is a sidechain of a genetically coded ammo
acid except glycine;
3
R is a C-terminal blocking group or from 0 to
4 amino acid residues, or -OH, or ORn where
Rn is straight or branched alkyl or cycloalkyl of 1 to 6 carbon atoms;
R 4 is a sidechain of a genetically coded ammo
acid, except glycine, or
-HC=CH2,
-OCH,
- 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.
Preferred compounds of the invention are those of Formula I selected from:
Formyl-MeMet-Leu-Phe-Obzl,
Formyl-Met-MeLeu-Phe-OBzl,
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-Pro-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,
MeGlp-His-Pro, Glp-MeHis - Pro,
Glp-His -MePro,
MeArg-Tyr-Leu-Pro-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,
Thr-Lys-MePro-Arg,
Thr-Lys-Pro-MeArg,
MeThr-Pro-Arg-Lys,
Thr-MePro-Arg-Lys,
Thr-Pro-MeArg-Lys,
Thr-Pro-Arg-MeLys,
MeThr-Val-Leu,
Thr-MeVal-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,
Arg-Pro-Tyr-Melle-Leu,
Arg-Pro-Tyr-Ile-MeLeu,
MeLeu-Asp-Ile-Ile-Trp, Leu-MeAsp-Ile-Ile-Trp,
Leu-Asp-MeIle-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-Gln,
Lys-Trp-MeAsp-Asn-Gln,
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,
Tyr-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,
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,
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 Nα-4-hydroxyphenylacetyl derivatives and the NH2-CO-(CH2)4-CO- derivatives of the above four compounds,
N-[[(4-Chlorophenyl)methoxy]carbonyl]-L-tryptophyl-α-methyl-DL-phenylalaninamide,
N-[[[4-(Trifluoromethyl)phenyl]methoxy]carbonyl]--L-tryptophyl-α-methyl-DL-phenylalaninamide,
N-[([1,1'-Biphenyl]-4-ylmethoxy)carbonyl]-L-tryptophyl-α-methyl-DL-phenylalaninamide,
N-[(9-Anthracenylmethoxy)carbonyl]-L-tryptophyl-α-methyl-DL-phenylalaninamide,
N-[(1-Naphthalenylmethoxy)carbonyl]-L-tryptophyl- α-methyl-DL-phenylalaninamide.
N-[(1-Naphthalenylmethoxy)carbonyl]-L-tryptophyl-α-methyl-L-phenylalaninamide,
N-[(1-Naphthalenylmethoxy)carbonyl]-L-tryptophyl-α-methyl-D-phenylalaninamide,
N-[[[4-(Propoxycarbonyl)phenyl]methoxy]carbonyl]-L-tryptophyl-α-methyl-DL-phenylalaninamide, N-[(Phenylmethoxy)carbonyl]-L-tryptophyl-α-methyl-DL-phenylalanylglycinamide,
N-[(1-Naphthalenylmethoxy)carbonyl]-L-tryptophyl-α-methyl-DL-phenylalanylglycinamide,
N-[(1-Naphthalenylmethoxy)carbonyl]-L-tryptophyl-α-methyl-L-phenylalanylglycinamide,
N-[[(2,3-Dimethoxyphenyl)methoxy]carbonyl]-L-tryptophyl-α-methyl-DL-phenylalaninamide,
N-[[(2,3-Dimethoxyphenyl)methoxy]carbonyl]-L-tryptophyl-α-methyl-L-phenylalanylglycinamide,
N-[(1-Naphthalenylmethoxy)carbonyl]-L-tryptophyl-α-methyl-DL-phenylalanyl-β-alaninamide,
N-[(1-Naphthalenylmethoxy)carbonyl]-L-tryptophyl-N-[2-acetylamino)ethyl]-α-methyl-DL-phenylalaninamide, N-[(1-Naphthalenylmethoxy)carbonyl]-L-tryptophyl-4-chloro-α-methyl-DL-phenylalanyl-glycinamide,
N-[(1-Naphthalenylmethoxy)carbonyl]-L-tryptophyl-α,4-dimethyl-DL-phenylalanyl-glycinamide,
N-[(1-Naphthalenylmethoxy)carbonyl]-L-tryptophyl-α-methyl-3-(2-thienyl)-DL-alanylglycinamide,
N-[[(4-Benzoylphenyl)methoxy]carbonyl]-L-tryptophyl-α-methyl-DL-phenylalaninamide,
N-[[(4-Nitrophenyl)methoxy]carbonyl]-L-tryptophyl-α-methyl-DL-phenylalaninamide,
α-Methyl-N-[N-[(1-naphthalenylmethoxy)carbonyl]-L-tryptophyl]-DL-phenylalanine-2-[(1,1-dimethylethoxy)-carbonyl]hydrazide, and
L-phenylalaninamide, N-[[(2,3-dimethoxyphenyl)-methoxy]carbonyl]-L-tryptophyl-N-(2-hydroxyethyl)-α-methyl.
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 recrystallization and reconversion to the nonsalt form.
The compounds of the present invention may be formed by coupling individual substituted α-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 Braña, 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 α-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) and Examples 21 (20), 28, 29. In Scheme 2 below, Compound 1 was activated with DCCI and HODhbt to give 14, which was reacted with GlyNH2 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 18-20
(17,18,19), and by a similar method to give
Examples 22-24.
Figure imgf000022_0001
In Scheme 3 below, AlaOMe was protected as its Schiffs base (1). The anion of (1) was then reached with 4-chlorobenzylbromide and deprotected to give (2). The amine (2) was then reacted with activated
1-Naphthmoc Trp to afford the dipeptide (3).
Hydrolysis of the methyl ester gave the acid (4), which was activated and reacted with GlyNH2 to give the final compound (5). (Example 25)
Figure imgf000024_0001
In Scheme 4 below, compound (1) was alkylated with 4-methylbenzyl bromide and deprotected to give (6). The amine (6) was then reacted with activated
1-Naphthmoc-Trp to afford the dipeptide (7).
Hydrolysis of the methyl ester gave the acid (8), which was activated and reacted with GlyNH2 to give compound (9). (Example 26)
Figure imgf000026_0001
In Scheme 5 below, 2-thienylalanine was protected as its methyl ester (10) and then as the Schiffs base (11). The anion of (11) was then alkylated with methyl iodine and deprotected to give the amine (12). The amine (12) was then reacted with activated
1-Naphthmoc-Trp to afford the dipeptide (13).
Hydrolysis of the methyl ester gave the acid (14), which was activated and reacted with GlyNH2 to give compound (15). (Example 27)
Figure imgf000028_0001
In Scheme 6 below, α-methyl-L-phenylalanine was activated with DCCI and HoDhbt and reacted with GlyNH2 in DMF to give the dipeptide 1. Removal of the BOC group with TFA gave the amine 2 which was reacted with BOC-(L)-Trp activated with DCCI and HoBT to give the tripeptide 3. The BOC group of 3 was subsequently removed with TFA to give the amine 4. This was treated with the appropriate mixed carbonate to give the desired compound 5 (Example 30).
Figure imgf000030_0001
In Scheme 7 below, BOC-α-methyl-L-phenylalanine was activated with HBTU and DIPEA and coupled with ethanol amine to give 1. The BOC protecting group was removed with TFA to give the free amine 2. The mixed carbonate of 2,3-dimethoxybenzylalcohol was reacted with (L)-tryptophan in the presence of base to give the 2,3-dimethoxybenzyloxycarbonyl-(L)-tryptophan 3. The acid 3 was activated with HBTU and DIPEA and reacted with the amine 2 to give the desired product 4
(Example 31).
Figure imgf000032_0001
For 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 which 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.
Some 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 Bi.ochem 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 × 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 peptide (Z-DPhe-Phe-Gly), DNA binding peptide
(Lys-Trp-Lys), Molluscan cardioexcitatory peptide
(Phe-Met-Arg-Phe-NH2 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-α-MePhe-LLeu (isomer 1) and
LTyr-Gly-Gly-α-MePhe-LLeu (isomer 2) whose full chemical names are N-[α-methyl-N-[N-(N-L-tyrosylglycyl)glycyl]-L-phenylalanyl]-L-leucine trifluoroacetate (1:1 salt) and N-[α-methyl-N-[N-(N-L-tyrosylglycyl)glycyl]-D-phenylalanyl]-L-leucine trifluoroacetate (1:1 salt).
Some of the compounds were evaluated in three tachykinin binding assays:
For the NK1 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).
Figure imgf000037_0001
Figure imgf000038_0001
Figure imgf000039_0001
The data in Tables III and III-A above show that the compounds are selective NK2 receptor ligands.
The compounds of Examples 18 and 19 illustrate NK2 receptor antagonism. See Table IV below. Therefore, they are expected to be useful in treating disorders mediated by tachykinins, eg, respiratory disorders, inflammation, gastrointestinal disorders, ophthalmic diseases, allergies, pain, circulatory insufficiency, diseases of the central nervous system, and migraine. TABLE IV. NK2 Functional Data Summary
Tissue Agonist Antagonist Cone. Dose\
Ratio pKa
Rat Colon Eledoisin Example 18 1 μM 3.1 6.5
Example 19 1 μM 7. 7 6.8
1,659,874 10 μM 10.3 7.0
Example 22 - - - - 7.9
Hamster Bladder
Urinary NKA Example 18 3 μM 30 7.0
Example 19 3 μM 29 6.9
1.659,874 3 μM 29 6.9
Hamster Trachea NKA Example 18 1 μM 11 6.9
Example 18 3 μM 46 7.2
Example 19 3 μM 31 6.9
Example 19 10 μM 472 7.2
L659,874 1 μM 11 7.0
L659,874 3 μM 41 7.1 a L659,874 is a standard NK2 antagonist. Its structure is
Glycinamide, N-acetyl-L-leucyl-L-methionyl-L-glutaminyl- L-tryptophyl-L-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.
For 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-αMePhe-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-αMePhe-LLeu NMR (D2O) δ 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-αMePhe-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-α-methylphenylalanine (1)
DL-α-Methylphenylalanine (5.0 g, 28 mmol) was dissolved in warm 10% Na2CO3 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 H2O. This was then washed with CH2Cl2 (3x), acidified with citric acid, and extracted with CH2Cl2 (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%): 1H NMR (DMSO-d6) δ 1.18 (3H, s), 1.41 (9H, s), 2.91 (1H, d, J 13 Hz), 3.31 ( 1H, d , J 13 Hz ) , 6 . 71 ( 1H, bs ) , 7 . 09 (2H , d) , 7 . 25 ( 3H m) , 12 . 50 ( 1H, bs ) .
EXAMPLE 3
N-(t-Butyloxycarbonyl)-DL-α-methylphenylalaninamide (2)
1 (6.7 g, 22 mmol) was dissolved in CH2Cl2 (80 mL) and 1,3-dicyclohexylcarbodiimide (4.9 g, 24 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 CH2Cl2. The aqueous layer was washed with CHCl2 (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 (1H, bs), 7.15 (6H, m), 7.41 (1H, bs).
EXAMPLE 4
DL-α-Methylphenylalaninamide (3)
2 (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%) 1H 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-ylmethoxy)carbonyl]-L-tryptophyl-DL-α-methylphenylalaninamide (4)
3 (1.8 g, 10.1 mmol) and N-[(9H-fluoren-9-ylmethoxy)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-α-methylphenylalaninamide (5)
4 (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 extracted 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%): 1Η NMR (DMSO-d6) δ 1.45
(0.5 × 3H, s), 1.51 (0.5 × 3H, s), 1.81 (2H, bs), 2.59 (0.5 × H, dd, J 10.14 Hz), 2.76 (0.5 × H, dd, 9,
14 Hz), 3.15 (2H, m), 3.45 (2H, m), 6.95-7.60 (12H, m), 8.16 (0.5 × H, s), 8.28 (0.5 × H, s), 10.85 (1H, s). EXAMPLE 7
N-[[(4-Chlorophenyl)methoxy]carbonyl]-L-tryptophyl-α-methyl-DL-phenylalaninamide (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 CH2Cl2 (5 mL) at 0°C and stirred for 5 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] : 1H NMR (DMSO-d6) δ 1.38 (0.5 × 3H, s, α-CH3), 1.40 (0.5 × 3H, s, α-CH3), 2.80-3.40 (4H, m, CH2-indole, CH2Ph), 4.10-4.32 (1H, m, Trp α-H), 4.80-4.99 (2H, m, CH2O), 6.90-7.78 (18H, m, Ar, CONH2, CONH, OCONH) , 10.70 (1H, s, indole NH); Anal. (C29H29N4O4C· 0.3 H2O) C, H, N.
EXAMPLE 8
N-[[[4-(Trifluoromethyl)phenyl]methoxy]carbonyl]-L-tryptophyl-α-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]; 1H NMR (DMSO-d6) δ 1.38 (0.5 × 3H, s, α-CH3), 1.41 (0.5 × 3H, s, α-CH3), 2.70, 3.46 (4H, m, CH2-indole, CH2Ph), 4.12-4.36 (1H, m, Trp α-H), 4.90-5.16 (2H, m, CH2O), 6.90-7.88 (18H, m, Ar, CONH2 CONH, OCONH), 10.84 (1H, s, indole NH); Anal. (C30H29N4O4F3- 0.75 H2O) C, H, N. EXAMPLE 9
N-[([1,1'-Biphenyl]-4-ylmethoxy)carbonyl]-L-tryptophyl-α-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) δ 1.38 (0.5 × 3H, s, α-CH3), 1.39 (0.5 × 3H, s, α-CH3), 2.72-3.40 (4H, m, CH2-indole, CH2Ph), 4.12-4.35 (1H, m, Trp α-H), 4.90-5.08 (2H, m, CH2O), 6.92-7.85 (23H, m, Ar, CONH2, CONH, OCONH), 10.90 (1H, s, indole NH); Anal. (C35H34N4O4-0.75 H2O) C, H, N.
EXAMPLE 10
N-[(9-Anthracenylmethoxy)carbonyl]-L-tryptophyl-α-methyl-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]; 1H NMR (DMSO-d6) 1.36 (3H, s, α-CH3), 2.25-3.20 (2H, m, CH2-indole), 3.25-3.40 (2H, m, CH2Ph), 4.20-4.38 (1H, m, Trp α-H), 5.95 (0.5 × 2H, AB, J 12 Hz, CH2Ar), 6.01 (0.5 × 2H, AB, J 12 Hz, CH2Ar), 6.85-7.20 (17H, m, Ar, CONH2, OCONH), 7.77 (0.5 × H, s, CONH), 7.83 (0.5 × H, S, CONH), 8.10 (1H, S, Ar), 8.13 (1H, S, Ar), 8.28 (1H, s, Ar), 8.31 (1H, s, Ar), 8.67 (1H, s, Ar), 10.80 (1H, s, indole NH); Anal. (C37H34N4O4 · 0.7 H20) C, H, N.
EXAMPLE 11
N-[(1-Naphthalenylmethoxy)carbonyl]-L-tryptophyl-α-methyl-DL-phenylalaninamide (10)
Prepared by the same method as 6. White foam (0.13 g, 34%); mp 102-113°C; MS (FAB) m/e 549 [M+H]; 1H NMR (DMSO-d6) δ 1.40 (3H, s), 2.87 (1H, m), 3.05-3.45 (3H, m), 4.15-4.40 (1H, m), 5.30-5.50 (2H, m), 6.90-8.00 (21H, m), 10.70 (1H, s); Anal. (C33H32N4O4- 0.4 H2O) C, H, N. EXAMPLE 12
N-[(1-Naphthalenylmethoxy)carbonyl]-L-tryptophyl-α-methyl-L-phenylalaninamide (11)
Prepared by the same method as 6, -α-methyl-L-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
N-[(1-Naphthalenylmethoxy)carbonyl]-L-tryptophyl-α-methyl-D-phenylalaninamide (12)
Prepared by the same method as 6. D-α-methylphenylalanine was obtained by the method of Turk, et al.
EXAMPLE 14
N-[[[4-(Propoxycarbσnyl)phenyllmethoxy]carbonyl]-L-tryptophyl-α-methyl-DL-phenylalaninamide (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 CH2Cl2 (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 H2O and 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 (95:5) gave 13 as a white foam (0.02 g, 13%); mp 78-105°C; MS (FAB) m/e 586 [M+H] ; 1H NMR (DMSO-d6) δ 0.97 (3H, t, J 7 Hz, CH3CH2), 1.38 (0.5 × 3H, s, α-CH3), 1.40 (0.5 × 3H, s, α-CH2), 1.76 (2H, m,
CH2CH2), 2.80-3.45 (4H, m, CH2-indole, CH2Ph), 4.12-4.35 (3H, m, Trp α-H, CH2CH2O), 4.94-5.15 (2H, m, CH2OCO), 6.95-7.95 (18H, m, Ar, CONH2, CONH, OCONH, 10.82 (1H, s, indole NH); Anal. (C33H36N4O6- 0.75 H2O) C, H, N. EXAMPLE 15
Boc(α-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-1,2,3-benzotriazine-4-(3H)-one (3.91 g,
24 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 white solid precipitated. This was filtered and dried to give 14 (9.1 g, 89%): 1H NMR (DMSO-d6) δ 1.44 (3H, S, α-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, 5, 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-(α-Me)-DLPheGlyNH2 (15)
14 (4.24 g, 10 mmol), glycinamide hydrochloride (1.11 g, 11 mmol), and triethylamine (1.39 mL, 11 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) δ 1.16 (3H, s, α-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
(α-Me)-DLPheGlyNH2 (16)
15 (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, α-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), 7.00-7.40 (7H, m, Ph2CONH2), 8.09 (3H, bs, NH3+), 8.62 (1H, d, J 5.6 Hz, CONH).
EXAMPLE 18
N-[(Phenylmethoxy)carbonyl]-L-tryptophyl-α-methyl-DL-phenylalanylqlycinamide (17)
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]; 1H NMR
(DMSO-d6) δ 1.19 (0.5 × 3H, s, α-CH3), 1.24 (0.5 × 3H, s, α-CH3), 2.80-3.30 (4H, m, CH2-indole, CH2Ph),
3.40-3.78 (2H, m, CH2CONH2), 4.25-4.48 (1H, m,
Trp-α-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 × H, t, CONHCH2) , 7.88 (0.5 × H, t, C0NHCH2) , 8.35 (0.5 × H, s, NHC(CH3) , 8.41 (0.5 × H, S, NHC(CH3) , 10.86 (1H, s, indole NH) ; Anal. (C31H33N5O35·0.5H2O) C,
H, N. EXAMPLE 19
N-[(1-Naphthalenylmethoxy)carbonyl]-L-tryptophyl-α-methyl-DL-phenylalanylglycinamide (18)
Diisopropylethylamine (52 μL, 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 temperature. 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 H2O (100 mL) and extracted with EtOAc (2 × 50 mL). The organic extracts were combined, washed with 10% citric acid solution (1 × 100 mL), saturated NaHCO3 solution (1 × 100 mL), H2O
(2 × 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]: 1H NMR (DMSO-d6) δ 1.21
(0.5 × 3H, s, α-CH3), 1.26 (0.5 × 3H, s, α-CH3),
2.85-3.78 (6H, m, CH2-indole, CH2Ph, Ch2CONH2), 4.38 (1H, m, Trp α-H), 5.44 (2H, m, CH2 naphthyl), 6.92-7.20 (10H, m, Ar/NH), 7.33 (0.5 × H, s, Ar/NH), 7.35
(0.5 × H, s, Ar/NH), 7.52-7.70 (6H, m, Ar/NH),
7.75-8.00 (4H, m, Ar/NH), 8.33 (0.5 × H, S, NH), 8.35 (0.5 × H, s, NH), 10.72 (1H, s, NH indole); Anal.
(C35H35N5O5-0.5H2O) C, H, N.
EXAMPLE 20
N-[(1-Naphthalenylmethoxy)carbonyl]-L-tryptophyl-α-methyl-L-phenylalanylqlycinamide (19)
Prepared by the same method as Compound 18 as in Example 19 from (L)α methylphenylalanine obtained by the method of Turk, et al. EXAMPLE 21
N-[[(2,3-Dimethoxyphenyl)methoxylcarbonyl]-L-tryptophyl-α-methyl-DL-phenylalaninamide (20)
Prepared by the same method as Compound 6 in Example 7, white foam: mp 87-102°C; MS (FAB) m/e 559 [M+H]; 1H NMR (DMSO-d6) δ 1.39 (3H, s, α-CH3), 2.88 (1H, t) and 3.11-3.39 (3H, m, CH2-indole, CH2Ph), 3.63 (3H, d, OCH3), 3.78 (3H, s, 0CH3), 4.21 (1H, m, Trp α-H), 4.93 (2H, s, CH2O), 6.79-7.78 (17H, m, Ar, CONH2, CONH, OCONH), 10.76 (1H, s, indole NH), and
(C31H34N4O6-0.5 H2O) C, H, N.
EXAMPLE 22
N-[[(2,3-dimethoxyphenyl)methoxylcarbonyl]-L-tryptophyl-α-methyl-L-phenylalanylqlycinamide
Prepared by a similar method as for Compound 19 in Example 20: white solid (0.100 g; 58%); MS (FAB) m/e 616 [M+H]; 1H NMR (DMSO-dg) δ 1.25 (3H, s, α-CH3), 2.92-3.35 (4H, m, CH2-indole, CH2Ph), 3.46-3.72 (5H, m, Gly CH2, OCH3), 3.78 (3H, s, OCH3), 4.28 (1H, m, Trp α-H), 4.87 (1H, d, J = 12.5 Hz, 1/2 Ar CH2O2CNH), 4.97 (1H, d, J = 12.5 Hz, 1/2 Ar CH2O2ONH) , 6.82-7.18 (13H, m, Ar, NH2), 7.33 (1H, d, J = 8 Hz, indole H),
7.55-7.63 (2H, m, NH, indole H), 7.85 (1H, bt, NH), 8.30 (1H, s, NH), 10.78 (1H, S, indole NH); Anal.
(C33H37N5O7-0.5 H2O) C, H, N.
EXAMPLE 23
N-[(1-naphthalenylmethoxy)carbonyl]-L-tryptophyl-α-methyl-DL-phenylalanyl-β-alaninamide
Prepared by a similar method as for Compound 18 in Example 19: colorless foam (0.062 g, 43%); MS (FAB) m/e 620.5 [M+H]; 1H NMR (DMSO-d6) δ 1.31 (3H, s, α-CH3), 2.22 (2.5H, m, 1/2 CHPh β-H + CH2CONH2), 2.71 (0.5 × H, d, J = 13 Hz, 1/2 CHPh β-H), 2.89 (1H, m,
CHPh β-H), 3.23 (4H, m, CH2-indole, CONHCH2), 4.26 (1H, m, Trp α-H), 5.43 (2H, m, CH2-naphthyl), 6.81 (1H, br.s, Ar/NH), 6.95-7.18 (8H, m, Ar/NH), 7.33 (2H, m, Ar/NH), 7.42-7.74 (7H, m, Ar/NH), 7.87-7.97 (4H, m, Ar/NH), 10.80 (1H, s, NH indole); Anal. (C36H37N5O5- 0.6 H2O) C, H, N.
EXAMPLE 24
N-[(1-naphthalenylmethoxy)carbonyl]-L-tryptophyl-N-[2 -acetylamino)ethyl]-α-methyl-DL-phenylalaninamide
Prepared by a similar method as for Compound 18 in Example 19: white solid MS (FAB) m/e 634 [M+H]; 1H NMR (DMSO-d6) δ 1.28 (3H, bs, α-CH3), 2.09 (3H, s, CH3CO), 2.3-3.8 (8H, m, NHCH2CH2NH, CH2-indole, CH2Ph), 4.32 (1H, m, Trp α-H), 5.43 (2H, m, CH2-naphthyl), 6.88-8.06 (21H, m, ArH, NH), 10.80 (1H, s, NH indole).
EXAMPLE 25
(See Scheme 3)
N-[(1-naphthalenylmethoxy)carbonyl]-L-tryptophyl-4-chloro-α-methyl-DL-phenylalanyl-glycinamide
Step 1
To a suspension of alanine methyl ester
hydrochloride (5 g) in CH2Cl2 (50 mL) was added
p-chlorobenzaldehyde (5.6 g) and MgSO4 (2 spatula loads) and NEt3 (4.02 g). The reaction mixture was stirred at room temperature for 3 days, filtered, and concentrated. The residue was taken up in Et2O, filtered again, and the filtrate concentrated to yield the desired Schiffs base (1) as a yellow oil (8.0 g, 99%); IR (film) 2953, 1742, and 1645 cm-1; 1H NMR
(CDCl3) 1.52 (3H, d, J = 7 Hz), 3.75 (3H, s), 7.37 (2H, d, J = 9 Hz), 7.70 (2H, d, J = 9 Hz), 8.27 (1H, s).
Step 2
To a solution of the Schiffs base (1) (1 g) in THF
(15 mL) at -78°C was added LHMDS (4.87 mL of 1 M) dropwise. After stirring for 1/2 hour at -78°C p-chlorobenzyl bromide (0.907 g) in THF (3 mL) was added and the reaction mixture allowed to warm to room temperature overnight. The reaction mixture was then concentrated and the residue stirred in an aqueous 1 M HCl solution (15 mL) for 4 hours. After this time the aqueous layer was washed with Et2O, made basic with a saturated NaHCO3 solution, and extracted with EtOAc. The organic layer was dried (MgSO4) and evaporated to yield the desired amine (2) as a yellow oil (0.77 g, 70%); IR (film) 3845 and 1734 cm-1; 1H NMR (CDCl3) 1.38 (3H, s), 1.58 (2H, brs, NH2), 2.76 (1H, d, 1/2 of ABq, J = 14 Hz), 3.07 (1H, d, 1/2 of ABq, J = 14 Hz), 3.70 (3H, s), 7.08 (2H, d, J = 8 Hz), 7.25 (2H, d,
J = 8 Hz).
Step 3
To a solution of 1-Naphthmoc-Trp (250 mg) in DMF (4 mL) was added the amine (Step 2) (146 mg) followed by BOP reagent (285 mg) and DIPEA (166 mg). The reaction mixture was stirred overnight at room
temperature and then evaporated to dryness. The residue was taken up in EtOAc and the organic layer washed with a 0.1 M HCl solution. The organic layer was then separated, dried (MgSO4), and evaporated to yield the crude product as a yellow oil (700 mg). The crude product was purified by column chromatography (eluent - 50% EtOAc/n-hexane) to yield the desired amide (3) as an amorphous white solid (295 mg, 77%); mp 72-75°C; IR (film) 3351, 1713, and 1668 cm-1; 1H NMR (CDCl3) 1.41 and 1.45 (1H, 2s), 3.00-3.40 (4H, m), 4.50 (1H, brs), 5.40 (1H, br.m), 5.60 (2H, s), 6.25 (1H, brs), 6.69 (1H, d, J = 9 Hz), 6.75 (2H, br.s),
6.90-7.10 (4H, m), 7.19 (1H, t, J = 8 Hz), 7.33 (1H, d, J = 8 Hz), 7.40-7.55 (4H, m), 7.70 (1H, br.m),
7 . 85 - 8 .05 (4H, m) ; Anal . (C34H33ClN3O5) C, H, N. Step 4
To a solution of the ester (Step 3) (270 mg) in THF (8 mL)/H2O (2 mL) was added LiOH (21 mg). The reaction mixture was stirred overnight at room
temperature and then evaporated to dryness. The residue was partitioned between aqueous 0.5 M HCl solution and EtOAc. The organic layer was separated, dried (MgSO4), and evaporated to yield the desired crude acid (4) as an amorphous white solid (237 mg, 90%); mp 80-85°C; IR (film) 3280, 1713, and 1665 cm-1; 1H NMR (CDCl3) 1.30 and 1.34 (1H, 2s), 2.95-3.20 (4H, m), 4.60 (1H, brm), 5.55 (2H, s), 5.65 (1H, br.s), 6.35 (1H, br.s), 6.68 (1H, brs), 6.70-6.95 (4H, m), 7.05 (2H, d, J = 7 Hz), 7.17 (1H, t, J = 7 Hz), 7.29 (1H, t, J = 7 Hz), 7.40-7.50 (5H, m), 7.65 (1H, brs), 7.80-8.10 (4H, m); Anal. (C33H30ClN3O5 · 0.5 H2O) C, H, N.
Step 5
To a solution of the acid (Step 4) (216 mg) in DMF (4 mL) was added the HCl-GlyNH2 (41 mg) followed by BOP reagent (164 mg) and DIPEA (144 mg). The reaction mixture was stirred overnight at room temperature and then evaporated to dryness. The residue was taken up in EtOAc and the organic layer washed with aqueous 0.1 M HCl solution. The organic layer was then
separated, dried MgSO4, and evaporated to yield the crude product (240 mg) as a brown gum. The crude product was purified by column chromatography
(eluent - 5% MeOH/CH2Cl2) to yield the desired
amide (5) as an amorphous white solid (190 mg, 80%); mp 100-105°C; IR (film) 3300 and 1664 cm1; 1H NMR
(CDCI3) 1.14 and 1.32 (1H, 2s), 2.65-3.25 (4H, m), 3.70 (2H, m), 4.30 (1H, m), 5.50 (2H, s), 5.55 (2H, m), 5.95 and 6.20 (1H, 2s), 6.70 (3H, m), 6.94 (2H, s),
6.95-7.15 (3H, m), 7.20 (1H, t, J = 7 Hz), 7.30-7.60
( 6H, m) , 7 . 80 (2H, m) , 7 .95 ( 1H, m) , 8 .28 and 8 .34 ( 1H, 2s) ; Anal . (C35H34ClN5O6 - 0 .5 H2O) C, H, N. EXAMPLE 26
(See Scheme 4)
N-[(1-naphthalenylmethoxy)carbonyl]-L-tryptophyl-α,4-dimethyl-DL-phenylalanyl-glycinamide
AlaOMe was protected as its Schiffs base (1). The anion of (1) was then reacted with 4-chlorobenzyl bromide and deprotected to give (2). The amine (2) was then reacted with activated Napthoc-Trp to afford the dipeptide (3). Hydrolysis of the methyl ester gave the acid (4), which was activated and reacted with GlyNH2 to give the title compound.
Step 1
Prepared by the same method as Compound 2 (in Example 25). Amine (6) colorless oil (310 mg, 68%); IR (film) 3370, 2950, 1734, and 1516 cm-1; 1E NMR (CDCl3) 1.38 (3H, s), 1.65 (2H, br.s), 2.73 (1H, d, 1/2 of ABq, J = 13 Hz), 3.06 (1H, d, 1/2 of ABq, J = 13 Hz), 3.70 (3H, s), 3.78 (3H, s), 6.81 (2H, d, J = 8 Hz), 7.06 (2H, d, J = 8 Hz).
Step 2
Prepared by the same method as Compound 3 (in Example 25). Amide (7) was produced as an amorphous white solid (113 mg, 31%); mp 65-70°C; IR (film) 3401, 1733, and 1666 cm-1; 1H NMR (CDCl3) 1.42 and 1.47 (3H, 2 singlets), 2.22 and 2.24 (3H, 2 singlets), 2.90-3.35 (4H, m), 3.66 (3H, s), 4.48 (1H, brs), 5.45 (1H, m), 5.57 (2H, s), 6.19 (1H, brs), 6.66 (2H, s), 6.73 (1H, d, J = 7 Hz), 6.85 (1H, d, J = 7 Hz), 6.93 (2H, d,
J = 7 Hz ) , 7. 10 (1H, brs) , 7 . 18 (1H, t , J = 7 Hz ) , 7.31 ( 1H, m) , 7.45 (4H, m) , 7 . 70 ( 1H, m) , 7. 80 - 8 . 10 (4H, m) ; Anal . (C35H35N3O5 - 0 .25 H2O) C, H, N. Step 3
Prepared by the same method as Compound 4 (in Example 25). Acid (8) was produced crude, without purification, as an amorphous white solid (85 mg, 93%); IR (film) 3350, 1713, and 1666 cm-1.
Step 4
Prepared by the same method as Compound 5 (in
Example 25). Amide (9) was produced as an amorphous white solid (56 mg, 67%); mp 90-95°C; IR (film) 3307 and 1662 cm-1; 1H NMR (CDCl3) 1.17 and 1.35 (3H,
2 singlets), 2.23 (3H, s), 2.63 (1/2H, d, 1/2 of ABq, J = 14 Hz), 2.80-3.25 (3 1/2H, m), 3.50-3.75 (2H, m),
4.36 (1H, m), 5.43 (2H, m) , 5.82 (1H, brs), 5.90 (1/2H, brs), 6.20 (1/2H, brs), 6.45 (1/2H, br.s), 6.55-6.80 (2 1/2H, m), 6.90-7.20 (6H, m), 7.30-7.60 (6H, m),
7.75-7.85 (2H, m), 7.95 (1H, t, J = 7 Hz), 8.71 (1H, brs); Anal. (C36H37N5O5-0.4 CHCl3) C, H, N.
EXAMPLE 27
(See Scheme 5)
N-[(1-naphthalenylmethoxy)carbonyl]-L-tryptophyl-α-methyl-3-(2-thienyl)-DL-alanylglycinamide
Step 1
To a suspension of S-2-thienylalanine (1 g) in MeOH (20 mL) was added SOCl2 (2.09 g). The reaction mixture was refluxed for 3 hours and then concentrated to yield a crude sample of the hydrochloride salt of the methyl ester 10. (The free amine was liberated upon wash up with a saturated NaHCO3/CH2Cl2
separation.) Hydrochloride salt 10 was produced as a white solid (1.24 g, 97%); 1H NMR (6D-DMSO) 3.41 (2H, d, J = 6 Hz), 3.72 (3H, s), 4.29 (1H, t, J = 6 Hz), 7.05 (2H, brs), 7.45 (1H, brs), 8.75 (2H, brs).
Step 2
Prepared by the same method as Compound 1 (in Example 25) Schiffs base (11) prepared as a crude sample and was a white solid (686 mg, 81%); IR (film) 2952, 1739, and 1642 cm-1; 1H NMR (CDCl3) 3.37 (1H, dd, J = 9 and 15 Hz), 3.56 (1H, dd, J = 5 and 15 Hz), 3.76 (3H, s), 4.16 (1H, dd, J = 5 and 9 Hz), 6.78 (1H, d, J = 3 Hz), 6.86 (1H, dd, J = 3 and 5 Hz), 7.09 (1H, dd, J = 1 and 5 Hz), 7.36 (2H, d, J = 9 Hz), 7.66 (2H, d, J = 9 Hz), 8.01 (1H, S).
Step 3
To a solution of the Schiffs base (11) (686 mg) in dry THF (10 mL) at -78°C was added LHMDS (2.5 mL of
1 M) dropwise. The reaction mixture was then stirred at -78°C for 1/2 hour and then Mel (426 mg) in THF (3 mL) added dropwise. The reaction was allowed to warm to room temperature over 4 hours and then an aqueous 1 M HCl solution (10 mL) added. After stirring overnight at room temperature, the reaction mixture was concentrated and the residue partitioned between water and Et2O. The aqueous layer was separated, made basic with saturated NaHCO3 solution, and extracted with EtOAc. The combined EtOAc extracts were dried (MgSO4) and evaporated to yield a crude sample of the
amine (12) as a yellow oil (195 mg, 42%); IR (film) 3400, 2951, and 1734 cm-1; 1H NMR (CDCl3) 1.40 (3H, s), 3.02 (1H, d, 1/2 of ABq, J = 14 Hz), 3.35 (1H, d, 1/2 of ABq, J = 14 Hz), 3.74 (3H, s), 6.83 (2H, d,
J = 3 Hz), 6.93 (2H, dd, J = 4 and 6 Hz), 7.15 (2H, d, J = 6 Hz).
Step 4
Prepared by the same method as Compound 3 (in
Example 25). Ester (13) was produced as an amorphous white solid (120 mg, 23%); mp 58-63°C; IR (film) 3338, 1721, 1711, and 1667 cm1; 1H NMR (CDCl3) 1.47 (3H, brs), 3.07 (1/2H, dd, J = 8 and 11 Hz), 3.20-3.35 (2 1/2H, m), 3.67 and 3.70 (4H, 2 singlets over broad multiplet), 4.55 (1H, brm), 5.45 (1H, m), 5.57 (2H, s), 6.37-6.95 (4H, m), 7.00-7.20 (3H, m), 7.32 (1H, d, J = 8 Hz), 7.35-7.55 (4H, m), 7.60-8.10 (4H, m);
Anal. (C32H31N3O5S) C, H, N. Step 5
Prepared by the same method as Compound 4 (in Example 25). Acid (14) was prepared as a crude sample without purification and was a white amorphous solid (90 mg, 93%); IR (film) 3340, 1721, 1715, and 1667 cm-1.
Step 6
Prepared by the same method as Compound 5 (in Example 25). Amide (15) was prepared as a white solid (40 mg, 43%); mp 98-102°C; IR (film) 3308, 3507, and 1662 cm"1 (broad); 1H NMR (CDCl3) 1.13 and 1.35 (3H, 2 singlets), 3.05-3.85 (6H, m), 4.45 (1H, m), 5.45 (2H, m), 5.80 (2H, m), 6.54 (2H, brs), 6.75-7.55 (12H, m), 7.70-8.00 (3H, m), 8.65 and 8.82 (1H, 2 singlets);
Anal. (C33H33N5O5S-0.5 CHCl3) C, H, N.
EXAMPLE 28
N-[[(4-benzoylphenyl)methoxy]carbonyl]-L-tryptophyl-α-methyl-DL-phenylalaninamide
Prepared by the same method as Compound 6 in
Example 7: white solid (0.100 g, 58%); MS (FAB) m/e 603 [M+H]; 1H NMR (DMSO-d6) δ 1.39 (3H, s, α-CH3), 2.89-3.38 (4H, m, CH2 indole, CH2Ph), 4.02 (1H, m, Trp αH), 5.07 (2H, m, Ar CH2O2CNH) , 6.97-7.85 (23H, m, Ar, NH, NH2, OCONH), 10.84 (1H, s, indole NH); Anal.
(C36H34N4O5-0.3 H2O) C, H, N. EXAMPLE 29
N-[[(4-nitrophenyl)methoxy]carbonyl]-L-tryptophyl-α-methyl-DL-phenylalaninamide
Prepared by the same method as Compound 6 in
Example 7: yellow foam (0.37 g, 68%); MS (FAB) m/e 544 [M+H]; 1H NMR (DMSO-d6) δ 1.39 (0.5 × 3H, S, α-CH3), 1.42 (0.5 × 3H, s, α-CH3), 3.16-3.40 (4H, m,
CH2 indole, CH2Ph), 4.22 (1H, m, Trp α-H), 5.10 (2H, m, ArCH2O2CNH), 6.95-8.16 (19, m, Ar, NH, NH2, OCONH), 10.80 (1H, s, indole NH); Anal. (C29H29N5O6- 0.25 H2O) C, H, N.
EXAMPLE 30
N-[[(2,3-dimethoxyphenyl)methoxy]carbonyl]- (L)-tryptophyl-α-methyl-(L)-phenylalanylglycinamide (5)
N-[[(1,1-dimethylethoxy)carbonyl]-α-methyl-(L)-phenyl¬alanyl-glycinamide (1)
1,3-Dicyclohexyxcarbondiimide (4.95 g, 24 mmol) was added to a solution of α-methyl-L-phenylalanine (obtained by the method of Turk, et al) (6.70 g,
24 mmol) in THF (100 mL) at -15°C and stirred for
5 minutes. 3-Hydroxy-1,2,3-benzotriazine-4-(3H)-one (3.91 g, 24 mmol) and an additional volume of THF
(20 mL) were added to the mixture at -10°C for 1 hour and at 0°C for 4 hours. After leaving overnight at 5°C, a white solid precipitated. This was filtered and dried to give the Dhbt ester of α-methylphenylalanine. The crude Dhbt ester (4.2 g, 10 mmol) and triethylamine (1.4 mL, 11 mmol) were dissolved in DMF (50 mL) and glycinamide hydrochloride (1.1 g, 11 mmol) added. The mixture was stirred at room temperature for 72 hours. The mixture was evaporated to dryness in vacuo and the residue taken up in EtOAc. This was washed with saturated aqueous NaHCO3, IN HCl solution and H2O, dried (MgSO4), and the solvent removed in vacuo to give 1 as a white solid (2.1 g, 62%), mp 78-82°C; 1H NMR (CDCI3): δ 1.38 (3H, S, CH3), 1.46 (9H, s, (CH3)3), 3.09 (1H, d, 13.8 Hz, CHHPh), 3.36 (1H, d, 13.8 Hz CHHpH) , 3.81 (1H, dd, 17.3, 5.7 Hz, CHH), 4.11 (1H, dd, 17.4, 7.0 Hz, CHH), 4.73 (1H, s, OCONH), 5.32 (1H, bs, CONHH), 6.62 (1H, bt, CONH), 7.10-7.25 (3H, m, ArH, CONHH), 7.25-7.35 (3H, m, Ar);
IR 3319, 1669, 1521 cm-1; MS (CI) 336 (M+H);
Anal. C17H25N3O4-0.25H2O (C,H,N). α-Methyl-(L)-phenylalanyl-glycinamide (2)
Trifluoroacetic acid (30 mL) was added dropwise to a stirred solution of 1 (3.7 g 11 mmol) in Ch2Cl2
(100 mL). The mixture was stirred at room temperature for 4 hours, the solvent removed in vacuo and the residue triturated with Et2O to give 2 as its TFA salt (3.7 g, 95%);
1H NMR (DMSO) : δ 1.52 (3H, S, CH3), 3.05 (1H, d,
14.1 Hz, CHHPh), 3.20 (1H, d, 14.0 Hz, CHHPh), 3.75 (2H, d, 5.6 Hz, CH2CO), 7.10 (1H, S, CONHH), 7.20-7.35 (6H, m, ArH, CONHH), 8.06 (3H, bs, NH3), 8.60 (1H, bt, CONH) ;
MS m/e (CI) 236 (100%) (M+H) ;
Anal. C12H17N3O2-C2HF3O2 (C,H,N). N-[(1,1-Dimethylethoxy)carbonyl]-(L)-tryptophyl-α-methyl-(L)-phenylanalyl-glycinamide (3)
N-[(1,1-Dimethylethoxy)carbonyl]-(L)-tryptophan (1.0 g, 3.3 mmol) was dissolved in DMF (20 mL) and 1-hydroxybenzotriazole hydrate (0.5 g, 3.7 mmol) added, followed by 1,3-dicyclohexylcarbodiimide (0.68 g,
3.3 mmol). The mixture was stirred at room temperature for 1 hour and then 2 (0.8 g, 3.4 mmol) in DMF (5 mL) was added. The mixture was stirred for a further
48 hours and the solvent removed in vacuo. The residue was taken up in EtOAc and filtered. Washed with saturated aqueous NaHCO3, IN HCl and H2O, dried
(MgSO4), and the solvent removed in vacuo to give a foam. This was purified by flash chromatography on silica eluting with EtOAc to give 3 as a white solid (1.25 g, 73%), mp 108-114°C;
1H NMR (CDCl3): δ 1.35 (9H, s, C(CH3)3), 1.45 (3H, s, CH3), 3.00 (1H, d, 13.7 Hz, CHHPh), 3.10-3.35 (3H, m, 3 × CHH), 3.77 (2H, d, 6.2 Hz, CH2), 4.15-4.25 (1H, m, CH), 5.13 (1H, s, NH), 5.48 (1H, s, NH), 6.29 (1H, s, NH), 6.95-7.00 (2H, m), 7.06 (1H, 2.2 Hz, ArH),
7.10-7.30 (7H, m), 7.41 (1H, d, 8.0 Hz, ArH), 7.56 (1H, d, 7.8 Hz, ArH), 8.38 (1H, s, indole NH);
IR 3310, 2980, 1653 cm"1;
MS (m/e) (FAB) 522 (M+H) ;
Anal. C28H35N5O5-0.5H2O (C,H,N). (L)-Tryptophyl-α-methyl-(L)-phenylalanyl-glycinamide (4)
3 (1.6 g, 3 mmol) was dissolved in CH3Cl2 (100 mL) and trifluoroacetic acid (50 mL) added. The mixture was stirred at room temperature for 2 hours and then the solvent removed in vacuo. The residue was
triturated with Et2O to give a solid. This was
partitioned between 10% aqueous NH3 solution and
CH2Cl2. The aqueous layer was separated and extracted with CH2Cl2 (2X). The organic extracts were combined, dried (MgSO4), and removed in vacuo. The solid was purified by flash chromatography on silica eluting with CH2Cl2:MeOH (10:1) to give 4 as a white solid (1.2 g, 93%), mp 105-108°C;
1H NMR (CDCl3): δ 1.46 (3H, s, CH3), 3.02-3.12 (2H, m, CHH, CHHPh), 3.21 (1H, dd, 14.4 Hz, CHH), 3.32 (1H, d, 13.7 Hz, CHHPh), 3.57 (1H, dd, 17.1, 5.9 Hz, CHH), 3.75-3.80 (1H, m, CH), 3.86 (1H, dd, 17.2, 6.7 Hz, CHH), 5.42 (1H, s, NHC), 6.14 (1H, bt, NHCH2),
7.05-7.10 (3H, m), 7.14 (1H, t, 7.0 Hz, ArH), 7.20-7.30 (5H, m, +CHCl3), 7.39 (1H, d, 8.0 Hz, ArH), 7.55 (1H, s, indole NH, CONHH), 7.64 (1H, d, 7.8 Hz, Ar), 8.40 (1H, s, indole NH); IR 3307, 1661 cm-1;
MS m/e (Cl) 442 (M+H) ;
Anal. C23H27N5O3-0.25H2 (C,H,N). N-[[(2,3-Dimethoxyphenyl)methoxy]carbonyl-(L)-tryptophyl-α-methyl-(L)-phenylalanyl-glycineamide (5) 2,3-Dimethoxybenzylalcohol (3.36 g, 20 mmol) and pyridine (1.7 mL, 20 mmol) were dissolved in CH2Cl2 and cooled to 0°C. 4-Nitrophenylchloroformate (4.8 g, 24 mmol) was dissolved in CH2Cl2 (50 mL) and added dropwise to the above over 15 minutes. The mixture was stirred at room temperature overnight, washed with H2O, dried (MgSO4) and the solvent removed in vacuo. The residue was triturated with Et2O to give the mixed carbonate as a crystalline solid (4.5 g, 68%),
mp 87-89°C;
1H NMR (CDCl3): δ 3.89 (3H, s, OCH3), 3.91 (3H, s, OCH3) , 5.37 (2H, S, CH2) 6.95-7.12 (3H, m, ArH), 7.39 (2H, d, 9.1 Hz, ArH), 8.27 (2H, d, 9.1 Hz, ArH);
IR 2940, 1766, 1525, 1486 cm-1;
MS m/e (Cl) 333 (M);
Anal. C23H27N5O3-0.25H2O (C,H,N).
The mixed carbonate (0.4 g, 1.2 mmol) described above, 4 (0.5 g, 1.2 mmol), and triethylamine (0.17 mL, 2.3 mmol) were dissolved in DMF (25 mL) and stirred at room temperature overnight. The solvent was removed in vacuo and the residue was taken up in EtOAc. This was washed with 10% aqueous Na2CO3 solution, IN HC1, 10% aqueous Na2CO3, dried (MgSO4), and the solvent removed in vacuo. The residue was purified by flash chromatography on silica eluting with a mixture of CH2Cl2:MeOH (9:1) to give 5 as a white solid (0.4 g, 55%), mp 106-110°C;
1Η NMR (dg-DMSO): δ 1.25 (3H, s, CH3), 2.85-2.95 (1H, m, CHH), 3.00-3.10 (2H, m, 2 × CHH), 3.30-3.40 (1H, obscured by water, CHH), 3.50 (1H, dd, 16.8, 5.4 Hz, CHH), 3.66 (3H, s, OCH3), 3.60-3.75 (1H, m, CHH), 3.80 (3H, S, OCH3), 4.25-4.35 (1H, m, CH), 4.88 (1H, d,
12.4 Hz, CHH), 4.99 (1H, d, 12.4 Hz, CHH), 6.80-6.85 (1H, m, ArH), 6.90-7.20 (12H, m, CONH2 indole (5C)H, (6C)H, (2C)H, and ArH), 7.25 (1H, d, 8.0 Hz, indole (7C)H), 7.58 (1H, d, 6.3 Hz, CONHCH), 7.63 (1H, d,
7.5 Hz, indole (4C)H), 7.87 (1H, bt, CONHCH2), 8.31 (1H, s, CONHC), 10.82 (1H, s, indole NH);
MS m/e (FAB+) 616 (70%) (M+H) + 638 (100%) (M+Na)+;
Anal . C33H37N5O7 - 0 . 5H2O ( C, H, N) .
EXAMPLE 31
L-phenylalaninamide, N-[[(2,3-dimethoxyphenyl)methoxy]-carbonyl]-L-tryptophyl-N-(2-hydroxyethyl)-α-methyl
N-[(2,3-Dimethoxyphenyl)methoxylcarbonyl]- (L)-tryptophyl-α-methyl-(L)-phenylalanyl-NH
Figure imgf000062_0001
OH
Step 1
Figure imgf000062_0002
Figure imgf000062_0003
N-BOC-α-methyl-(L)-tryptophyl-ethanol amide (1)
N-BOC-α-Me-(L)Phe(H)-DCHA (4.6 g, 10 mmol) was taken up in CH2Cl2 and washed with 1N HCl, H2O, dried (MgSO4), and the solvent was removed in vacuo to give the acid as a solid. The solid was taken up in DMF (50 mL) and stirred with HBTU (3.80 g, 10 mmol) and diisopropylethyl amine (2.58 g, 20 mmol) at room temperature for 30 minutes. The ethanol amine (0.61 g, 10 mmol) and diisopropylethylamine (1.29 g, 1 mmol) were added to the reaction mixture which was stirred at room temperature overnight. The solvent was removed and the residue was taken up in EtOAc and washed with IN HCl, saturated NaHCO3 (aqueous), water, and brine. The organic layer was dried (MgSO4) and the solvent was removed in vacuo to give 1 as a cream amorphous solid (2.98 g, 92%);
1H NMR (CDCl3): δ 1.39 (3H, s, α-Me), 1.47 (9H, s, CH3)C), 3.10 (1H, d, 13.7 Hz, PhCHH), 3.34 (1H, d, 13.7 Hz, PhCHH), 3.2-3.6 (2H, m, CONHCHH), 3.6-3.85 (2H, m, CHHOH), 4.74 (1H, s, CONH(CH2) 2OH), 6.52 (1H, m, urethane NH), 7.13 (2H, dd, 1.4, 7.3 Hz, ArH),
7.26-7.31 (3H, m, ArH).
Step 2
Figure imgf000063_0001
α-Methyl-(L)-tryptophyl-ethanol amide (2)
1 (2.98 g, 9.3 mmol) was stirred in a solution of CH2Cl2:TFA (40 mL:25 mL) for 2 hours. The reaction mixture was washed with Na2CO3 solution, H2O, dried
(MgSO4), and concentrated in vacuo to give 2 as a clear oil (1.64 g, 79%). This was used crude for the next reaction. Step 3
Figure imgf000064_0001
N-[[(2,3-Dimethoxyphenyl)methoxylcarbonyl]- (L)-tryptophan (3)
Figure imgf000064_0002
(3.33 g, 10 mmol), (L)-tryptophan (2.04 g,
10 mmol), triethylamine (2 mL) and DMF were stirred at room temperature over the weekend. The reaction mixture was concentrated in vacuo and the residue was taken up in CH2Cl2, washed (H2O), dried (MgSO4), and the solvent removed in vacuo to give a solid. This was purified by column chromatography on silica eluting with 2% MeOH:CH2Cl2 to give 3 as an amorphous solid; 1H NMR (CDCI3): δ 3.32 (2H, m, CH2 indole), 3.78 (3H, s, PhOMe), 3.84 (3H, s, PhOMe), 4.69 (1H, m, αH), 5.16 (2H, q, 12.4, 15.6 Hz, OCH2Ph), 6.89-7.10 (5H, m, 4Ar, CONH), 7.16 (1H, t, 7.2 Hz, ArH), 7.31 (1H, d, 8.3 Hz, H1-indole), 7.55 (1H, d, 7.4 Hz, H4-indole), 8.0 (1H, s, H2-indole), 8.1 (1H, bs, NH indole). Step 4
Figure imgf000065_0001
N-[[(2,3-Dimethoxyphenyl)methoxy]carbonyl]-(L)-tryptophyl-α-methyphenylalanyl-ethanol amide (4)
3 (2.95 g, 7.4 mmol), HBTU (2.81 g, 7.4 mmol) and diisopropylethylamine (1.90 g, 14.7 mmol) was stirred at room temperature in DMF (40 mL) for 30 minutes.
Diisopropylethylamine (0.95 g, 7.4 mmol) and a solution of 2 (1.64 g, 7.4 mmol) in DMF (10 mL) were added to the mixture which was stirred at room temperature
overnight. The solvent was removed and the residue was taken up in EtOAc and washed with 1N HCl, saturated
NaHCO3 (aqueous), brine, and water. The organic
solution was dried (MgSO4) and the solvent was removed in vacuo to give a solid which was purified by flash chromatography on silica eluting with 10% MeOH:DCM to give 4 as a cream solid (1.96 g, %), mp 76-79°C;
1H NMR (CDCI3): δ 1.48 (3H, S, αMe), 2.84 (1H, d,
13.6 Hz, OH), 3.16-3.30 (6H, m, CHH indole,
NHCHHCHHOH), 3.62 (2H, m, CH3CCHHPh), 3.78 (3H, s,
CH3OPh), 3.84 (3H, s, CH3OPh), 4.24 (1H, q, αH), 5.02
(2H, dd, 12.0, 25.5 Hz, OCHHPh), 5.32 (1H, d, 4.9 Hz,
NHCO2), 6.12 (1H, s, CONH), 6.82 (1H, bs, CONHCH2),
6.86-7.39 (11H, m, ArH), 7.38 (1H, d, 8.1 Hz, ArH),
7.60 (1H, d, 7.7 Hz, ArH), 8.25 (1H, S, indole NH);
IR 3332, 1649, 1277 cm-1;
Anal. C33H38N4O7-0.6H2) (C,H,N).

Claims

1. A compound of Formula
Figure imgf000066_0001
or a pharmaceutically acceptable salt thereof wherein:
R1 is an N-terminal blocking group, from 0 to
4 amino acid residues or hydrogen;
R 2 is a sidecham of a genetically coded ammo acid except glycine;
R 3 is a C-terminal blocking group from 0 to
4 amino acid residues, -OH, or ORn wherein Rn is straight or branched alkyl or cycloalkyl of from 1 to 6 carbon atoms;
R 4 is a sidecham of a genetically coded ammo acid, except glycine, or
-HC=CH2,
-C≡CH,
-CH2 -CH=CH2 ,
-CH2C≡CH,
-CH2Ar,
-CH2OR,
-CH2OAr,
- (CH2)nCO2R, or
- (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;
R4 and R2 cannot be hydrogen; R1 and R3 together cannot be more than 4 amino acid residues.
2. A compound according to Claim 1 wherein:
R1 is H,
Boc,
Fmoc,
Z,
1-Adoc,
2-Adoc,
IVA, or
NVA;
R2 is CH3- or
HOOC-CH2- ; and
R3 is -NH2,
-OCH3, or
-OCH2Ph.
3. A compound according to Claim 1 selected from MeLys-Trp-Asp-Asn-Gln,
Lys-MeTrp-Asp-Asn-Gln,
Lys-Trp-MeAsp-Asn-Gln,
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,
Tyr- 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,
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,
MePhe-Phe-Gly-Leu-Met-NH2,
Phe-MePhe-Gly-Leu-Met-NH2,
Phe-Phe-Gly-MeLeu-Met-NH2, and
Phe-Phe-Gly-Leu-MeMet-NH2.
4. A compound named N-[α-methyl-N-[N-(N-L- tyrosylglycyl)glycyl]-L-phenylalanyl]-L-leucine trifluoroacetate (1:1 salt).
5. A compound named N-[α-methyl-N-[N-(N-L- tyrosylglycyl)glycyl]-D-phenylalanyl]-L-leucine trifluoroacetate (1:1 salt).
6. A compound named N-[[(4-chlorophenyl)methoxy]- carbonyl]-L-tryptophyl-α-methyl-DL- phenylalaninamide.
7. A compound named N-[[[4-(trifluoromethyl)phenyl]- methoxy]carbonyl]-L-tryptophyl-α-methyl-DL- phenylalaninamide.
8. A compound named N-[([1,1'-biphenyl]-4-yl- methoxy)carbonyl]-L-tryptophyl-α-methyl-DL- phenylalaninamide.
9. A compound named N-[(9-anthracenylmethoxy)- carbonyl]-L-tryptophyl-α-methyl-DL- phenylalaninamide.
10. A compound named N-[(1-naphthalenylmethoxy)- carbonyl]-L-tryptophyl-α-methyl-DL- phenylalaninamide.
11. A compound named N-[(1-naphthalenylmethoxy)- carbonyl]-L-tryptophyl-α-methyl-L- phenylalaninamide.
12. A compound named N-[(1-naphthalenylmethoxy)- carbonyl]-L-tryptophyl-α-methyl-D- phenylalaninamide.
13. A compound named N-[[[4-(propoxycarbonyl)- phenyl]methoxy]carbonyl]-L-tryptophyl-α-methyl-DL- phenylalaninamide.
14. A compound named N-[(phenylmethoxy)carbonyl]-L- tryptophyl-α-methyl-DL-phenylalanylglycinamide.
15. A compound named N-[(1-naphthalenylmethoxy)- carbonyl]-L-tryptophyl-α-methyl-DL-phenylalanyl- glycinamide.
16. A compound named N-[(1-naphthalenylmethoxy)- carbonyl]-L-tryptophyl-α-methyl-L- phenylalanylglycinamide.
17. A compound named N-[[(2,3-dimethoxyphenyl)- methoxy]carbonyl]-L-tryptophyl-α-methyl-DL- phenylalaninamide.
18. A compound named N-[[(4-benzoylphenyl)- methoxy]carbonyl]-L-tryptophyl-α-methyl-DL- phenylalaninamide.
19. A compound named N-[[(4-nitrophenyl)- methoxy]carbonyl]-L-tryptophyl-α-methyl-DL- phenylalaninamide.
20. A compound named N-[[(2,3-dimethoxyphenyl)- methoxy]carbonyl]-L-tryptophyl-α-methyl-L- phenylalanylglycinamide.
21. A compound named N-[(1-naphthalenylmethoxy)- carbonyl]-L-tryptophyl-α-methyl-DL-phenylalanyl- β-alaninamide.
22. A compound named N-[(1-naphthalenylmethoxy)- carbonyl]-L-tryptophyl-N-[2-acetylamino)ethyl]-α- methyl-DL-phenylalaninamide.
23. A compound named N-[(1-naphthalenylmethoxy)- carbonyl]-L-tryptophyl-4-chloro-α-methyl-DL- phenylalanyl-glycinamide.
24. A compound named N-[(1-naphthalenylmethoxy)- carbonyl]-L-tryptophyl-α,4-dimethyl-DL- phenylalanyl-glycinamide.
25. A compound named N-[(1-naphthalenylmethoxy)- carbonyl]-L-tryptophyl-α-methyl-3-(2-thienyl)-DL- alanylglycinamide.
26. A compuond named L-phenylalaninamide,
N-[[(2,3-dimethoxyphenyl)methoxy]carbonyl]- L-tryptophyl-N-(2-hydroxyethyl)-α-methyl.
27. 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 carrier.
28. A method of treating pain in a mammal comprising administering an effective pain treating amount of a compound according to Claim 1.
PCT/US1993/009809 1992-10-19 1993-10-14 Analogues of cholecystokinin (30-33) containing an alpha-substituted aminoacid WO1994009031A1 (en)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2735687A1 (en) * 1995-06-21 1996-12-27 Sederma Sa Topical penta:peptide-contg. compsns. for slimming treatment
US8362068B2 (en) 2009-12-18 2013-01-29 Idenix Pharmaceuticals, Inc. 5,5-fused arylene or heteroarylene hepatitis C virus inhibitors
WO2015083816A1 (en) * 2013-12-06 2015-06-11 国立大学法人京都大学 New nk3 receptor agonist

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1992011284A1 (en) * 1990-12-17 1992-07-09 The James Black Foundation Limited Tetrapeptide derivatives and analogues
WO1992019254A1 (en) * 1991-04-24 1992-11-12 Warner-Lambert Company α-SUBSTITUTED POLYPEPTIDES HAVING THERAPEUTIC ACTIVITY

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1992011284A1 (en) * 1990-12-17 1992-07-09 The James Black Foundation Limited Tetrapeptide derivatives and analogues
WO1992019254A1 (en) * 1991-04-24 1992-11-12 Warner-Lambert Company α-SUBSTITUTED POLYPEPTIDES HAVING THERAPEUTIC ACTIVITY

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
D C HORWELL ET AL.: "alpha-methyl tryptophanylphenylalanines and their arylethylamine "dipeptoid" analogues of the tetrapeptide cholecystokinin (30-33)", EUR J MED CHEM, vol. 25, no. 1, February 1990 (1990-02-01), ELSEVIER, PARIS, pages 53 - 60, XP023870633, DOI: doi:10.1016/0223-5234(90)90164-X *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2735687A1 (en) * 1995-06-21 1996-12-27 Sederma Sa Topical penta:peptide-contg. compsns. for slimming treatment
US8362068B2 (en) 2009-12-18 2013-01-29 Idenix Pharmaceuticals, Inc. 5,5-fused arylene or heteroarylene hepatitis C virus inhibitors
US9187496B2 (en) 2009-12-18 2015-11-17 Idenix Pharmaceuticals Llc 5,5-fused arylene or heteroarylene hepatitis C virus inhibitors
WO2015083816A1 (en) * 2013-12-06 2015-06-11 国立大学法人京都大学 New nk3 receptor agonist

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