WO1993021219A1 - Endothelin antagonists ii - Google Patents

Endothelin antagonists ii Download PDF

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Publication number
WO1993021219A1
WO1993021219A1 PCT/US1993/003658 US9303658W WO9321219A1 WO 1993021219 A1 WO1993021219 A1 WO 1993021219A1 US 9303658 W US9303658 W US 9303658W WO 9321219 A1 WO9321219 A1 WO 9321219A1
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ile
trp
asp
leu
fmoc
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PCT/US1993/003658
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French (fr)
Inventor
Wayne Livingston Cody
Annette Marian Doherty
John Gordon Topliss
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Warner-Lambert Company
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Application filed by Warner-Lambert Company filed Critical Warner-Lambert Company
Priority to EP93912310A priority Critical patent/EP0647236A1/en
Priority to AU42904/93A priority patent/AU678357B2/en
Priority to JP5518657A priority patent/JPH07505890A/en
Priority to SK1287-94A priority patent/SK128794A3/en
Priority to KR1019940703745A priority patent/KR950701344A/en
Priority to CA002133090A priority patent/CA2133090A1/en
Priority to RU9494046056A priority patent/RU2100369C1/en
Publication of WO1993021219A1 publication Critical patent/WO1993021219A1/en
Priority to FI944905A priority patent/FI944905A0/en
Priority to NO944013A priority patent/NO944013L/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/575—Hormones
    • C07K14/57536—Endothelin, vasoactive intestinal contractor [VIC]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/02—Bacterial antigens
    • A61K39/09—Lactobacillales, e.g. aerococcus, enterococcus, lactobacillus, lactococcus, streptococcus
    • A61K39/092—Streptococcus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/02—Bacterial antigens
    • A61K39/095—Neisseria
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/02—Drugs for disorders of the urinary system of urine or of the urinary tract, e.g. urine acidifiers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/04—Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/08—Vasodilators for multiple indications
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/12—Antihypertensives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/60—Medicinal preparations containing antigens or antibodies characteristics by the carrier linked to the antigen
    • A61K2039/6031—Proteins
    • A61K2039/6068—Other bacterial proteins, e.g. OMP
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/62—Medicinal preparations containing antigens or antibodies characterised by the link between antigen and carrier
    • A61K2039/627—Medicinal preparations containing antigens or antibodies characterised by the link between antigen and carrier characterised by the linker
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the present invention relates to novel
  • novel compounds of the present invention are antagonists of endothelin useful in treating elevated levels of endothelin, acute and chronic renal failure, hypertension, myocardial infarction, metabolic, endocrinological and
  • neurological disorders congestive heart failure, endotoxic shock, subarachnoid hemorrhage, arrhythmias, asthma, preeclampsia, atherosclerotic disorders including Raynaud's disease, restenosis, angina, cancer, pulmonary hypertension, ischemic disease, gastric mucosal damage, hemorrhagic shock, ischemic bowel disease, and diabetes.
  • Endothelin-1 (ET-1), a potent vasoconstrictor, is a 21 amino acid bicyclic peptide that was first isolated from cultured porcine aortic endothelial cells. Endothelin-1, is one of a family of
  • bicyclic peptides which include; ET-2, ET-3, vasoactive intestinal contractor (VIC), and the sarafotoxins (SRTXs).
  • VOC vasoactive intestinal contractor
  • SRTXs sarafotoxins
  • ET-1 disulfide bridges of ET-1, which are the same for the endothelins, VIC, and the sarafotoxins, has led to significant speculation as to the importance of the resulting induced secondary structure to receptor binding and functional activity.
  • the flexible C-terminal hexapeptide of ET-1 has been shown to be important for binding to the ET receptor and
  • Trp-21 C-terminal amino acid
  • Endothelin is involved in many human disease states.
  • ET may be involved in the pathogenesis of congestive heart failure and myocardial ischemia
  • the effectiveness and specificity of the anti-ET antibody were confirmed by its capacity to prevent renal deterioration caused by a single bolus dose (150 pmol) of synthetic ET, but not by infusion of angiotensin II, norepinephrine, or the thromboxane A 2 mimetic U-46619 in isolated kidneys (Perico, N., et al, "Endothelin Mediates the Renal Vasoconstriction Induced by Cyclosporine in the Rat," J. Am. Soc.
  • SHR hypertensive rats
  • MAP mean arterial pressure
  • WKY normotensive Wistar-Kyoto rats
  • ET A and ET B The distribution of the two cloned receptor subtypes, termed ET A and ET B , have been studied extensively (Arai, H., et al, Nature 348:730 (1990), Sakurai, T., et al, Nature 348:732 (1990)).
  • the ET A or vascular smooth muscle receptor, is widely used.
  • N-Acetyl-ET[10-21,11,15-Ala] caused vasorelaxation in isolated, endothelium-intact porcine pulmonary
  • ET analogs are potent vasoconstrictors in the rabbit pulmonary artery, a tissue that appears to possess an ET B y, nonselective type of receptor (ibid).
  • Plasma endothelin-1 levels were dramatically increased in a patient with malignant
  • the ET receptor antagonist BQ-123 has been shown to block ET-1 induced bronchoconstriction and tracheal smooth muscle contraction in allergic sheep providing evidence for expected efficacy in bronchopulmonary diseases such as asthma (Noguchi, et al, Am. Rev.
  • Circulating endothelin levels are elevated in women with preeclampsia and correlate closely with serum uric acid levels and measures of renal
  • Plasma immunoreactive endothelin-1 concentrations are elevated in patients with sepsis and correlate with the degree of illness and depression of cardiac output (Pittett J., et al, Ann Sur ⁇ .. 1991, 213(3), 262).
  • ET-1 antagonist BQ-123 has been evaluated in a mouse model of endotoxic shock.
  • This ET A antagonist significantly increased the survival rate in this model (Toshiaki M., et al, 20.12.90.
  • Endothelin is a potent agonist in the liver eliciting both sustained vasoconstriction of the hepatic vasculature and a significant increase in hepatic glucose output (Gandhi C.B., et al, Journal of Biological Chemistry. 1990, 265(29), 17432).
  • streptozotocin-diabetic rats there is an increased sensitivity to endothelin-1 (Tammesild P.J., et al, Clin. Exp. Pharmacol. Physiol.. 1992, 19(4), 261).
  • increased levels of plasma ET-1 have been observed in microalbuminuric insulin-dependent
  • diabetes mellitus patients indicating a role for ET in endocrine disorders such as diabetes (Collier A., et al. Diabetes Care, 1992, 15(8), 1038).
  • ET A antagonist receptor blockade has been found, to produce an antihypertensive effect in normal to low renin models of hypertension with a time course similar to the inhibition of ET-1 pressor responses (Basil M.K., et al, J. Hypertension. 1992, 10(Suppl 4), S49).
  • the endothelins have been shown to be arrhythmogenic, and to have positive chronotropic and inotropic effects, thus ET receptor blockade would be expected to be useful in arrhythmia and other cardiovascular disorders (Han S.-P., et al, Life Sci., 1990, 46, 767).
  • ETs in neurological disorders.
  • the potent vasoconstrictor action of ETs on isolated cerebral arterioles suggests the importance of these peptides in the regulation of cerebrovascular tone. Increased ET levels have been reported in some CNS disorders, i.e., in the CSF of patients with
  • ET-1 induced lesion development in a new model of local ischemia in the brain has been
  • PTCA percutaneous transluminal coronary angioplasty
  • Elevated levels of endothelin have also been measured in patients suffering from ischemic heart disease (M. Yasuda, et al, Amer. Heart J.. 1990, 119 801-806, S.G. Ray, et al, Br. Heart J.. 1992, 67, 383-386) and either stable or unstable angina (J.T.
  • endothelin has been suggested that the proliferative effect of endothelin on mesangial ceils may be a contributing factor in chronic renal failure (P.J. Schultz, J. Lab. Clin. Med. , 1992, 119, 448- 449).
  • Local intra-arterial administration of endothelin has been shown to induce small intestinal mucosal damage in rats in a dose-dependent manner (S. Mirua, et al, Digestion. 1991, 48, 163-172).
  • endothelin-1 in the range of 50-500 pmol/kg into the left gastric artery increased the tissue type plasminogen activator release and platelet activating formation, and induced gastric mucosal hemorrhagic change in a dose dependent manner (I. Kurose, et al, Gut. 1992, 33, 868-871). Furthermore, it has been shown that an anti-ET-1 antibody reduced ethanol-induced vasoconstriction in a concentration-dependent manner (E. Masuda, et al, Am. J. Physiol., 1992, 262, G785-G790). Elevated endothelin levels have been observed in patients suffering from Crohn's disease and ulcerative colitis (S.H. Murch, et al, Lancet, 1992, 339, 381-384).
  • Serial Number 07/995,480 discloses a series of novel antagonists of endothelin.
  • the present invention is a compound of Formula I
  • R is hydrogen
  • R 9 is F, Cl, Br, or I
  • R 2a and R 3 are as defined above excluding R 3 is hydrogen, or , wherein R 2 is as defined above,
  • R 1 is hydrogen or alkyl
  • Z is
  • n is zero or an integer of 1 or 2
  • n is zero or an integer of 1, 2, 3, or 4,
  • n is as defined above, ,
  • R 1 and R 2 are as defined above, or ,
  • R 2 and R 3 are each the same or different and each is as defined above,
  • X and Y are the same and substituted at the same position on the aromatic ring and each may be one, two, three, or four substituents selected from the group consisting of hydrogen,
  • R 2 and R 3 are as defined above, or nitro or
  • R, Z, X, and Y are as defined above;
  • R 2b and R 3b are each the same or different and each is
  • R 2b is as defined above, , wherein R 2b and R 3b are each the same or different and each is as defined above for R 2b and R 3b , , wherein R 2b is as defined
  • R 1 and n are as defined above, or
  • AA 2 is absent
  • R 2b and R 3b are each the same or
  • R 2b is as defined above, or , wherein R 2b is as
  • R 1 and n are as defined above, or AA 3 is absent;
  • AA 4 and AA 5 are each independently absent or each is independently
  • R 6 is hydrogen
  • R 1 and n are as defined above;
  • R 8 is , wherein R 1 is as defined
  • R 1 is as defined above, or
  • R 1 and n are as defined above;
  • stereochemistry at C in AA 6 is L; or a
  • the compounds of Formula I are useful in the treatment of hypertension,
  • myocardial infarction myocardial infarction, metabolic, endocrinological and neurological disorders, congestive heart failure, endotoxic shock, subarachnoid hemorrhage, arrhythmias, asthma, and chronic and acute renal failure,
  • Atherosclerotic disorders including Raynaud's disease, restenosis, angina, cancer, pulmonary hypertension, ischemic disease, gastric mucosal damage, hemorrhagic shock, ischemic bowel disease, and diabetes.
  • invention is a pharmaceutical composition for
  • alkyl means a straight or branched hydrocarbon radical having from 1 to 12 carbon atoms and includes, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, undecyl, dodecyl, and the like.
  • alkenyl means a straight or branched unsaturated hydrocarbon radical having from 2 to
  • alkynyl means a straight or branched triple bonded unsaturated hydrocarbon radical having from 2 to 12 carbon atoms and includes, for example, ethynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 3-pentynyl, 1-hexynyl, 2-hexynyl,
  • cycloalkyl means a saturated
  • hydrocarbon ring which contains from 3 to 12 carbon atoms, for example, cyclopropyl, cyclobutyl,
  • cycloalkylalkyl means a saturated hydrocarbon ring attached to an alkyl group wherein alkyl is as defined above.
  • the saturated hydrocarbon ring contains from 3 to 12 carbon atoms. Examples of such are cyclopropylmethyl, cyclopentylmethyl, cyclohexylmethyl, adamantylmethyl and the like.
  • alkoxy and thioalkoxy are O-alkyl or S-alkyl as defined above for alkyl.
  • aryl means an aromatic radical which is a phenyl group, a benzyl group, a naphthyl group, a biphenyl group, a pyrenyl group, an anthracenyl group,
  • alkyl as defined above, alkoxy as defined above, thioalkoxy as defined above, hydroxy, thiol, nitro, halogen, amino, wherein alkyl is as defined above, wherein alkyl is as defined above, wherein alkyl is as defined above, wherein alkyl is
  • arylalkyl means an aromatic radical attached to an alkyl radical wherein aryl and alkyl are as defined above for example benzyl,
  • heteroaryl means a heteroaromatic radical which is 2-or 3-thienyl, 2- or 3-furanyl, 2-or 3-pyrrolyl, 2-, 4-, or 5-imidazolyl, 3-, 4-, or 5-pyrazolyl, 2-, 4-, or 5-thiazolyl, 3-, 4-, or
  • 5-isothiazolyl 2-, 4-, or 5-oxazolyl, 3-, 4-, or 5-isoxazolyl, 3- or 5-1,2,4-triazolyl, 4- or
  • heterocycloalkyl means 2- or
  • Halogen is fluorine, chlorine, bromine or iodine.
  • the compounds of Formula I are capable of further forming both pharmaceutically acceptable acid addition and/or base salts. All of these forms are within the scope of the present invention.
  • Pharmaceutically acceptable acid addition salts of the compounds of Formula I include salts derived from nontoxic inorganic acids such as hydrochloric, nitric, phosphoric, sulfuric, hydrobromic, hydriodic, hydrofluoric, phosphorous, and the like, as well as the salts derived from nontoxic organic acids, such as aliphatic mono- and dicarboxylic acids,
  • Such salts thus include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, trifluoroacetate, propionate, caprylate, isobutyrate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, mandelate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, phthalate, benzenesulfonate,
  • toluenesulfonate phenylacetate, citrate, lactate, maleate, tartrate, methanesulfonate, and the like.
  • salts of amino acids such as arginate and the like and gluconate, galacturonate (see, for example, Berge, S. M., et al,
  • the acid addition salts of said basic compounds are prepared by contacting the free base form with a sufficient amount of the desired acid to produce the salt in the conventional manner.
  • a peptide of Formula I can be converted to an acidic salt by treating with an aqueous solution of the desired acid, such that the resulting pH is less than 4.
  • the solution can be passed through a C18 cartridge to absorb the peptide, washed with copious amounts of water, the peptide eluted with a polar organic solvent such as, for example, methanol, acetonitrile, aqueous mixtures thereof, and the like, and isolated by concentrating under reduced pressure followed by lyophilization.
  • the free base form may be regenerated by contacting the salt form with a base and isolating the free base in the conventional manner.
  • the free base forms differ from their respective salt forms somewhat in certain physical properties such as solubility in polar solvents, but otherwise the salts are equivalent to their respective free base for purposes of the present invention.
  • Pharmaceutically acceptable base addition salts are formed with metals or amines, such as alkali and alkaline earth metals or organic amines.
  • metals or amines such as alkali and alkaline earth metals or organic amines. Examples of metals used as cations are sodium, potassium,
  • suitable amines are N,N' -dibenzylethylenediamine, chloroprocaine, choline, diethanolamine,
  • the base addition salts of said acidic compounds are prepared by contacting the free acid form with a sufficient amount of the desired base to produce the salt in the conventional manner.
  • a peptide of Formula I can be converted to a base salt by treating with an aqueous solution of the desired base, such that the resulting pH is greater than 9.
  • the solution can be passed through a C18 cartridge to absorb the peptide, washed with copious amounts of water, the peptide eluted with a polar organic solvent such as, for example, methanol, acetonitrile, aqueous mixtures thereof, and the like, and isolated by concentrating under reduced pressure followed by lyophilization.
  • the free acid form may be regenerated by contacting the salt form with an acid and isolating the free acid in the conventional manner.
  • the free acid forms differ from their respective salt forms somewhat in certain physical properties such as solubility in polar solvents, but otherwise the salts are equivalent to their respective free acid for purposes of the present invention.
  • Certain of the compounds of the present invention can exist in unsolvated forms as well as solvated forms, including hydrated forms.
  • the solvated forms, including hydrated forms are
  • Certain of the compounds of the present invention possess one or more chiral centers and each center may exist in the R(D) or S(L) configuration.
  • the present invention includes all enantiomeric and epimeric forms as well as the appropriate mixtures thereof.
  • a preferred compound of Formula I is one wherein AA 1 is
  • R 2 and R 3 are each the same or different and each is
  • R 2 and R 3 are as defined above.
  • R 9 is F, Cl, Br, or I, , wherei.n R 3 is as defined
  • R 3 is as defined above excluding R 3 is hydrogen
  • n is zero or an integer of 1 or 2
  • n is zero or an integer of
  • R 1 is hydrogen or alkyl
  • R 2 and R 3 are each the same or different and each is as defined above and X and Y are the same and substituted at the same position on the aromatic ring and each substituent is selected from the group consisting of
  • R 4 is hydrogen
  • R 2b and R 3b are each the same or different and each is
  • R 2b and R 3b are each the same or different and each is as defined above for
  • R 2b and R 3b wherein R 2b is as defined above, , wherein R 2b is as
  • R 2b is as
  • n is as defined above or
  • AA 2 is absent :
  • R 5 is aryl
  • R 2b and R 3b are each the same or different and each is as defined above, , wherein R 2b is as defined
  • n is as defined above, or
  • AA 3 is absent
  • AA 4 and AA 5 are each independently absent or each is
  • n is as defined above;
  • R 7 is aryl
  • n is as defined above, or
  • stereochemistry at CH in AA 6 is L; or a
  • a more preferred compound of Formula I is one wherein AA 1 is
  • R 2 and R 3 are each the same or different and each is
  • R 9 is F, Cl, Br, or I
  • arylalkyl excluding R 10 is hydrogen
  • n is zero or an integer
  • n is zero or an integer of
  • X and Y are each the same and substituted at the same position on the aromatic ring and each substituent is selected from the group consisting of
  • R 4 is hydrogen
  • R 2b and R 3b are each the same or
  • R 2b and R 3b are each the same or
  • R 2b is as defined above, or , wherein R 2b is as
  • n is an integer of 1, 2, 3, or 4 or
  • AA 2 is absent
  • AA 3 is , wherein R 3 is
  • n is an integer of 1, 2, 3, or 4;
  • AA 4 and AA 5 are each independently
  • R 6 is hydrogen
  • n is an integer of 1, 2, 3, or 4;
  • R 7 is aryl or heteroaryl
  • n is zero or an integer of 1, 2, 3, or 4, or
  • R 7 , R 1 , and n are as
  • stereochemistry at CH in AA 6 is L; or a
  • the cells used were rabbit renal artery vascular smooth muscle cells grown in a 48-well dish (1 cm 2 ) (confluent cells).
  • the growth media was Dulbecco's Modified
  • Eagles/Ham's F12 which contained 10% fetal bovine serum and antibiotics (penicillin/streptomycin/ fungizone).
  • the assay buffer was a medium 199 containing
  • the tissue is made up of 20 mM tris (hydroxy-methyl)aminomethane hydrochloride (Trizma) buffer, 2 mM ethylenediaminetetra acetate, 100 ⁇ M
  • Trizma tris (hydroxy-methyl)aminomethane hydrochloride
  • wash filters with an additional 2.5 mL of cold wash buffer.
  • Antagonist activity is measured by the ability of added compounds to reduce endothelin-stimulated arachidonic acid release in cultured vascular smooth muscle cells as arachidonic acid release (AAR).
  • the compounds of Formula I may be prepared by solid phase peptide synthesis on a peptide
  • synthesizer for example, an Applied Biosystems 430A peptide synthesizer using activated esters or
  • N-alpha-Boc protected amino acids on PAM or MBHA resins. Additionally, the compounds of Formula I may also be prepared by conventional solution peptide synthesis. Amino acid side chains are protected as follows: BzKAsp, Glu, Ser),
  • Each peptide resin (1.0 g) is cleaved with 9 mL of HF and 1 mL of anisole or p-cresol as a scavenger (60 minutes, 0°C). The peptide resin is washed with cyclohexane, extracted with 30% aqueous HOAc, followed by glacial HOAc, concentrated under reduced pressure, and lyophilized.
  • FAB-MS bombardment mass spectrometry
  • the compounds of the present invention can be prepared and administered in a wide variety of oral and parenteral dosage forms.
  • the compounds of the present invention can be administered by
  • the compounds of the present invention can be administered by inhalation, for example, intranasally. Additionally, the compounds of the present invention can be administered
  • dosage forms may comprise as the active component, either a compound of
  • pharmaceutically acceptable carriers can be either solid or liquid.
  • Solid form preparations include powders, tablets, pills, capsules, cachets,
  • a solid carrier can be one or more substances which may also act as diluents, flavoring agents, binders,
  • preservatives for example, preservatives, tablet disintegrating agents, or 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 properties in suitable proportions and compacted in the shape and size desired.
  • the powders and tablets preferably contain from five or ten to about seventy percent of the active compound.
  • Suitable carriers are magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, a low melting wax, cocoa butter, and the like.
  • the term "preparation" is intended to include the formulation of the active compound 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.
  • cachets and lozenges are included. Tablets, powders, capsules, pills, cachets, and lozenges can be used as solid dosage forms suitable for oral administration.
  • a low melting wax such as a mixture of fatty acid glycerides or cocoa butter
  • the active component is dispersed homogeneously therein, as by stirring.
  • the molten homogenous mixture is then poured into
  • Liquid form preparations include solutions, suspensions, and emulsions, for example, water or water propylene glycol solutions.
  • liquid preparations can be formulated in solution in aqueous polyethylene glycol solution.
  • Aqueous solutions suitable for oral use can be prepared by dissolving the active component in water and adding suitable colorants, flavors, stabilizing and thickening agents as desired.
  • Aqueous suspensions suitable for oral use can be made by dispersing the finely divided active component in water with viscous material, such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, and other well-known viscous material, such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, and other well-known viscous material, such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, and other well-known
  • solid form preparations which are intended to be converted, shortly before use, to liquid form preparations for oral administration.
  • Such liquid forms include solutions, suspensions, and emulsions. These preparations may contain, in
  • sweeteners such as sweeteners, dispersants, thickeners, solubilizing agents, and the like.
  • the pharmaceutical preparation is preferably in unit dosage form.
  • the preparation is subdivided 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 preparation, such as packeted tablets, capsules, and powders in vials or ampoules.
  • the unit dosage form can be a
  • capsules tablet, cachet, or lozenge itself, or it can be the appropriate number of any of these in packaged form.
  • the quantity of active component in a unit dose preparation may be varied or adjusted from 0.1 mg to 100 mg preferably 0.5 mg to 100 mg according to the particular application and the potency of the active component.
  • the composition can, if desired, also contain other compatible therapeutic agents.
  • the compounds utilized in the pharmaceutical method of this invention are administered at the initial dosage of about 0.01 mg to about 20 mg per kilogram daily.
  • a daily dose range of about 0.01 mg to about 10 mg per kilogram is preferred.
  • the dosages may be varied depending upon the requirements of the patient, the severity of the condition being treated, and the compound being employed. Determination of the proper dosage for a particular situation is within the skill of the art. Generally, treatment is initiated with smaller dosages which are less than the optimum dose of the compound. Thereafter, the dosage is increased by small increments until the optimum effect under the circumstances is reached. For convenience, the total daily dosage may be divided and administered in portions during the day, if desired.
  • the linear hexapeptide is prepared by standard solid phase synthetic peptide methodology utilizing a Boc/benzyl strategy (Stewart, J. M. and Young, J. D., Solid Phase Peptide Synthesis. Pierce Chemical Co., Rockford, IL, 1984). All protected amino acids and reagents are obtained from commercial sources with the exception of N- ⁇ -Boc-DL-Bhg and are not further purified.
  • the protected peptide resin is prepared on an Applied Biosystems 430A Peptide Synthesizer, utilizing protocols supplied for a dicyclohexyl-carbodiimide-mediated coupling scheme (Standard 1.0, Version 1.40). Starting with 0.710 g of
  • N- ⁇ -Boc-Trp-PAM resin (0.70 meq/g, 0.497 meq of
  • Boc-Trp(For) total the protected peptide is prepared by the stepwise coupling of the following amino acids (in order of addition): N- ⁇ -Boc-Ile.0.5H 2 O,
  • a typical cycle for the coupling of an individual amino acid residue is illustrated below (reproduced from the ABI manual):
  • the peptide is liberated from the solid support, and the carboxylate of aspartic acid deprotected by treatment with anhydrous hydrogen fluoride (9.0 mL), anisole (0.5 mL), and dimethyl sulfide (0.5 mL)
  • a saturated solution of sodium bicarbonate in water is prepared, diluted with water (1:10), chilled to 0°C, and 10 mL of the solution is added to
  • Bhg-HCl (1.70 g, 5.43 mmol) is suspended in

Abstract

Novel antagonists of endothelin are described, as well as methods for the preparation and pharmaceutical compositions of the same, which are useful in treating elevated levels of endothelin, acute and chronic renal failure, hypertension, myocardial infarction, metabolic, endocrinological, neurological disorders, congestive heart failure, endotoxic shock, subarachnoid hemorrhage, arrhythmias, asthma, preeclampsia, atherosclerotic disorders including Raynaud's disease, restenosis, angina, cancer, pulmonary hypertension, ischemic disease, gastric mucosal damage, hemorrhagic shock, ischemic bowel disease, and diabetes. In the claimed new compounds, the N-terminal 16-His is replaced by a building-block of formula (α).

Description

ENDOTHELIN ANTAGONISTS II
BACKGROUND OF THE INVENTION
The present invention relates to novel
antagonists of endothelin useful as pharmaceutical agents, to methods for their production, to
pharmaceutical compositions which include these compounds and a pharmaceutically acceptable carrier, and to pharmaceutical methods of treatment. More particularly, the novel compounds of the present invention are antagonists of endothelin useful in treating elevated levels of endothelin, acute and chronic renal failure, hypertension, myocardial infarction, metabolic, endocrinological and
neurological disorders, congestive heart failure, endotoxic shock, subarachnoid hemorrhage, arrhythmias, asthma, preeclampsia, atherosclerotic disorders including Raynaud's disease, restenosis, angina, cancer, pulmonary hypertension, ischemic disease, gastric mucosal damage, hemorrhagic shock, ischemic bowel disease, and diabetes.
Endothelin-1 (ET-1), a potent vasoconstrictor, is a 21 amino acid bicyclic peptide that was first isolated from cultured porcine aortic endothelial cells. Endothelin-1, is one of a family of
structurally similar bicyclic peptides which include; ET-2, ET-3, vasoactive intestinal contractor (VIC), and the sarafotoxins (SRTXs). The unique bicyclic structure and corresponding arrangement of the
disulfide bridges of ET-1, which are the same for the endothelins, VIC, and the sarafotoxins, has led to significant speculation as to the importance of the resulting induced secondary structure to receptor binding and functional activity. ET-1 analogues with incorrect disulfide pairings exhibit at least 100-fold less vasoconstrictor activity. The flexible C-terminal hexapeptide of ET-1 has been shown to be important for binding to the ET receptor and
functional activity in selected tissues.
Additionally, the C-terminal amino acid (Trp-21) has a critical role in binding and vasoconstrictor activity, since ET[1-20] exhibits approximately 1000-fold less functional activity.
Endothelin is involved in many human disease states.
Several in vivo studies with ET antibodies have been reported in disease models. Left coronary artery ligation and reperfusion to induce myocardial
infarction in the rat heart, caused a four- to
sevenfold increase in endogenous endothelin levels. Administration of ET antibody was reported to reduce the size of the infarction in a dose-dependent manner (Watanabe, T., et al, "Endothelin in Myocardial
Infarction," Nature (Lond.) 344:114 (1990)). Thus, ET may be involved in the pathogenesis of congestive heart failure and myocardial ischemia
(Margulies, K.B., et al, "Increased Endothelin in Experimental Heart Failure," Circulation 82:2226
(1990)).
Studies by Kon and colleagues using anti-ET antibodies in an ischemic kidney model, to deactivate endogenous ET, indicated the peptide's involvement in acute renal ischemic injury (Kon, V., et al,
"Glomerular Actions of Endothelin In Vivo," J. Clin. Invest. 83:1762 (1989)). In isolated kidneys, preexposed to specific antiendothelin antibody and then challenged with cyclosporine, the renal perfusate flow and glomerular filtration rate increased, while renal resistance decreased as compared with isolated kidneys preexposed to a nonimmunized rabbit serum. The effectiveness and specificity of the anti-ET antibody were confirmed by its capacity to prevent renal deterioration caused by a single bolus dose (150 pmol) of synthetic ET, but not by infusion of angiotensin II, norepinephrine, or the thromboxane A2 mimetic U-46619 in isolated kidneys (Perico, N., et al, "Endothelin Mediates the Renal Vasoconstriction Induced by Cyclosporine in the Rat," J. Am. Soc.
Nephrol. 1:76 (1990)).
Others have reported inhibition of ET-1 or ET-2-induced vasoconstriction in rat isolated thoracic aorta using a monoclonal antibody to ET-1 (Koshi, T., et al, "Inhibition of Endothelin (ET)-1 and ET-2-Induced Vasoconstriction by Anti-ET-1 Monoclonal
Antibody," Chem. Pharm. Bull., 39:1295 (1991)).
Combined administration of ET-1 and ET-1 antibody to rabbits showed significant inhibition of the blood pressure (BP) and renal blood flow responses
(Miyamori, I., et al, Systemic and Regional Effects of Endothelin in Rabbits: Effects of Endothelin
Antibody," Clin. Exp. Pharmacol. Physiol.. 17:691 (1990)).
Other investigators have reported that infusion of ET-specific antibodies into spontaneously
hypertensive rats (SHR) decreased mean arterial pressure (MAP), and increased glomerular filtration rate and renal blood flow. In the control study with normotensive Wistar-Kyoto rats (WKY) there were no significant changes in these parameters (Ohno, A.
Effects of Endothelin-Specific Antibodies and
Endothelin in Spontaneously Hypertensive Rats,"
J. Tokyo Women's Med. Coll.. 61:951 (1991)).
In addition, elevated levels of endothelin have been reported in several disease states (see Table I below).
Burnett and co-workers recently demonstrated that exogenous infusion of ET (2.5 ng/kg/mL) to
anesthetized dogs, producing a doubling of the
circulating concentration, did have biological actions (Lerman, A., et al, "Endothelin has Biological Actions at Pathophysiological Concentrations," Circulation
83:1808 (1991)). Thus heart rate and cardiac output decreased in association with increased renal and systemic vascular resistances and antinatriuresis. These studies support a role for endothelin in the regulation of cardiovascular, renal, and endocrine function.
In the anesthetized dog with congestive heart failure, a significant two- to threefold elevation of circulating ET levels has been reported (Cavero, P.G., et al, "Endothelin in Experimental Congestive Heart Failure in the Anesthetized Dog," Am. J. Physiol.
259:F312 (1990)), and studies in humans have shown similar increases (Rodeheffer, R.J., et al,
"Circulating Plasma Endothelin Correlates With the Severity of Congestive Heart Failure in Humans,"
Am. J. Hypertension 4:9A (1991)). When ET was
chronically infused into male rats, to determine whether a long-term increase in circulating ET levels would cause a sustained elevation in mean arterial blood pressure, significant, sustained, and dose- dependent increases in mean arterial BP were observed. Similar results were observed with ET-3 although larger doses were required (Mortenson, L.H., et al, "Chronic Hypertension Produced by Infusion of
Endothelin in Rats," Hypertension. 15:729 (1990)).
The distribution of the two cloned receptor subtypes, termed ETA and ETB, have been studied extensively (Arai, H., et al, Nature 348:730 (1990), Sakurai, T., et al, Nature 348:732 (1990)). The ETA, or vascular smooth muscle receptor, is widely
distributed in cardiovascular tissues and in certain regions of the brain (Lin, H.Y., et- al, Proc. Natl. Acad. Sci. 88:3185 (1991)). The ETB receptor,
originally cloned from rat lung, has been found in rat cerebellum and in endothelial cells, although it is not known if the ETB receptors are the same from these sources. The human ET receptor subtypes have been cloned and expressed (Sakamoto, A., et al, Biochem. Biophys. Res. Chem. 178:656 (1991), Hosoda, K., et al, FEBS Lett. 287:23 (1991)). The ETA receptor clearly mediates vasoconstriction and there have been a few reports implicating the ETB receptor in the initial vasodilatory response to ET (Takayanagi, R., et al, FEBS Lett. 282:103 (1991)). However, recent data has shown that the ETB receptor can also mediate
vasoconstriction in some tissue beds (Panek, R.L., et al, Biochem. Biophys. Res. Commun. 183(2):566
(1992)).
Comparison of the receptor affinities of the ETs and SRTXs in rats and atria (ETA) or cerebellum and hippocampus (ETB), indicate that SRTX-c is a selective ETB ligand (Williams, D.L., et al, Biochem. Biophys. Res . Commun.. 175:556 (1991)). A recent study showed that selective ETB agonists caused only vasodilation in the rat aortic ring, possibly through the release of EDRF from the endothelium (ibid). Thus, reported selective ETB agonists, for example, the linear analog ET [1, 3 , 11 , 15-Ala] and truncated analogs ET [6-21 ,
1,3,11,15-Ala], ET[8-21,11,15-Ala], and
N-Acetyl-ET[10-21,11,15-Ala] caused vasorelaxation in isolated, endothelium-intact porcine pulmonary
arteries (Saeki, T., et al, Biochem. Biophys. Res.
Commun. 179:286 (1991)). However, some ET analogs are potent vasoconstrictors in the rabbit pulmonary artery, a tissue that appears to possess an ETB y, nonselective type of receptor (ibid).
Plasma endothelin-1 levels were dramatically increased in a patient with malignant
hemangioendothelioma (K. Nakagawa et al, Nippon Hifuka Gakkai Zasshi. 1990, 100, 1453-1456).
The ET receptor antagonist BQ-123 has been shown to block ET-1 induced bronchoconstriction and tracheal smooth muscle contraction in allergic sheep providing evidence for expected efficacy in bronchopulmonary diseases such as asthma (Noguchi, et al, Am. Rev.
Respir. Pis.. 1992, 145 (4 Part 2), A858).
Circulating endothelin levels are elevated in women with preeclampsia and correlate closely with serum uric acid levels and measures of renal
dysfunction. These observations indicate a role for ET in renal constriction in preeclampsia (Clark B.A.., et al. Am. J. Obstet. Gvnecol., 1992, 166, 962-968).
Plasma immunoreactive endothelin-1 concentrations are elevated in patients with sepsis and correlate with the degree of illness and depression of cardiac output (Pittett J., et al, Ann Surσ.. 1991, 213(3), 262).
In addition the ET-1 antagonist BQ-123 has been evaluated in a mouse model of endotoxic shock. This ETA antagonist significantly increased the survival rate in this model (Toshiaki M., et al, 20.12.90.
EP 0 436 189 Al).
Endothelin is a potent agonist in the liver eliciting both sustained vasoconstriction of the hepatic vasculature and a significant increase in hepatic glucose output (Gandhi C.B., et al, Journal of Biological Chemistry. 1990, 265(29), 17432). In streptozotocin-diabetic rats there is an increased sensitivity to endothelin-1 (Tammesild P.J., et al, Clin. Exp. Pharmacol. Physiol.. 1992, 19(4), 261). In addition increased levels of plasma ET-1 have been observed in microalbuminuric insulin-dependent
diabetes mellitus patients indicating a role for ET in endocrine disorders such as diabetes (Collier A., et al. Diabetes Care, 1992, 15(8), 1038).
ETA antagonist receptor blockade has been found, to produce an antihypertensive effect in normal to low renin models of hypertension with a time course similar to the inhibition of ET-1 pressor responses (Basil M.K., et al, J. Hypertension. 1992, 10(Suppl 4), S49). The endothelins have been shown to be arrhythmogenic, and to have positive chronotropic and inotropic effects, thus ET receptor blockade would be expected to be useful in arrhythmia and other cardiovascular disorders (Han S.-P., et al, Life Sci., 1990, 46, 767).
The widespread localization of the endothelins and their receptors in the central nervous system and cerebrovascular circulation has been described
(Nikolov R.K., et al, Drugs of Today. 1992, 28(5),
303-310). Intracerebroventricular administration of ET-1 in rats has been shown to evoke several
behavioral effects. These factors strongly suggest a role for the ETs in neurological disorders. The potent vasoconstrictor action of ETs on isolated cerebral arterioles suggests the importance of these peptides in the regulation of cerebrovascular tone. Increased ET levels have been reported in some CNS disorders, i.e., in the CSF of patients with
subarachnoid hemorrhage and in the plasma of women with preeclampsia. Stimulation with ET-3 under conditions of hypoglycemia have been shown to
accelerate the development of striatal damage as a result of an influx of extracellular calcium.
Circulating or locally produced ET has been suggested to contribute to regulation of brain fluid balance through effects on the choroid plexus and CSF
production. ET-1 induced lesion development in a new model of local ischemia in the brain has been
described.
Circulating and tissue endothelin
immunoreactivity is increased more than twofold in patients with advanced atherosclerosis (A. Lerman, et al, New England J. Med.. 1991, 325, 997-1001).
Increased endothelin immunoreactivity has also been associated with Buerger's disease (K. Kanno, et al, J. Amer. Med. Assoc. 1990, 264, 2868) and Raynaud's phenomenon (M.R. Zamora, et al, Lancet, 1990, 336, 1144-1147). Likewise, increased endothelin
concentrations were observed in hypercholesterolemic rats (T. Horio, et al, Atherosclerosis. 1991, 89, 239-245).
An increase of circulating endothelin levels was observed in patients that underwent percutaneous transluminal coronary angioplasty (PTCA) (A. Tahara, et al, Metab. Clin. Eχp., 1991, 40, 1235-1237, K.
Sanjay, et al, Circulation. 1991, 84(Suppl. 4), 726).
Increased plasma levels of endothelin have been measured in rats (T.J. Stelzner, et al. Am. J.
Physiol.. 1992, 262, L614-L620) and individuals
(T. Miyauchi, et al, Jpn. J. Pharmacol.. 1992, 58, 279P, D.J. Stewart, et al, Ann. Internal Medicine.
1991, 114 464-469 ) with pulmonary hypertension.
Elevated levels of endothelin have also been measured in patients suffering from ischemic heart disease (M. Yasuda, et al, Amer. Heart J.. 1990, 119 801-806, S.G. Ray, et al, Br. Heart J.. 1992, 67, 383-386) and either stable or unstable angina (J.T.
Stewart, et al, Br. Heart J.. 1991, 66, 7-9).
Infusion of an endothelin antibody lh prior to and lh after a 60 minute period of renal ischaemia resulted in changes in renal function versus control. In addition, an increase in glomerular platelet-activating factor was attributed to endothelin (A.
Lopez-Farre, et al, J. Physiology. 1991, 444, 513- 522). In patients with chronic renal failure as well as in patients on regular hemodialysis treatment mean plasma endothelin levels were significantly increased
(F. Stockenhuber, et al, Clin. Sci. (Lond.). 1992, 82,
255-258). In addition it has been suggested that the proliferative effect of endothelin on mesangial ceils may be a contributing factor in chronic renal failure (P.J. Schultz, J. Lab. Clin. Med. , 1992, 119, 448- 449). Local intra-arterial administration of endothelin has been shown to induce small intestinal mucosal damage in rats in a dose-dependent manner (S. Mirua, et al, Digestion. 1991, 48, 163-172). Administration of endothelin-1 in the range of 50-500 pmol/kg into the left gastric artery increased the tissue type plasminogen activator release and platelet activating formation, and induced gastric mucosal hemorrhagic change in a dose dependent manner (I. Kurose, et al, Gut. 1992, 33, 868-871). Furthermore, it has been shown that an anti-ET-1 antibody reduced ethanol-induced vasoconstriction in a concentration-dependent manner (E. Masuda, et al, Am. J. Physiol., 1992, 262, G785-G790). Elevated endothelin levels have been observed in patients suffering from Crohn's disease and ulcerative colitis (S.H. Murch, et al, Lancet, 1992, 339, 381-384).
Recently the nonpeptide endothelin antagonist RO 46-2005 has been reported to be effective in models of acute renal ischemia and subarachnoid hemorrhage in rats (3rd International Conference on Endothelin, Houston, Texas, February 1993). In addition, the ETA antagonist BQ-123 has been shown to prevent early cerebral vasospasm following subarachnoid hemorrhage (M. Clozel and H. Watanabe, Life Sci.. 52: 825-834 (1993)).
TABLE I. Plasma Concentrations of ET-1 in Humans
ET Plasma
Condition Normal Levels Reported
Control
(pg/mL)
Atherosclerosis 1.4 3.2 pmol/L
Surgical operation 1.5 7.3
Buerger's disease 1.6 4.8
Takayasu's arteritis 1.6 5.3
Cardiogenic shock 0.3 3.7
Congestive heart failure (CHF) 9.7 20.4
Mild CHF 7.1 11.1
Severe CHF 7.1 13.8
Dilated cardiomyopathy 1.6 7.1
Preeclampsia 10.4 pmol/L 22.6 pmol/L
Pulmonary hypertension 1.45 3.5
Acute myocardial infarction 1.5 3.3
(several reports) 6.0 11.0
0.76 4.95
0.50 3.8
Subarachnoid hemorrhage 0.4 2.2
Crohn's Disease 0-24 fmol/mg 4-64 fmol/mg
Ulcerative colitis 0-24 fmol/mg 20-50 fmol/mg
Cold pressor test 1.2 8.4
Raynaud's phenomenon 1.7 5.3
Raynaud's/hand cooling 2.8 5.0
Hemodialysis <7 10.9
(several reports) 1.88 4.59
Chronic renal failure 1.88 10.1
Acute renal failure 1.5 10.4
Uremia before hemodialysis 0.96 1.49
Uremia after hemodialysis 0.96 2.19
Essential hypertension 18.5 33.9
Sepsis syndrome 6.1 19.9
Postoperative cardiac 6.1 11.9
Inflammatory arthritides 1.5 4.2
Malignant hemangioendothelioma 4.3 16.2
(after
removal)
Rovero, P., et al, British Journal of
Pharmacology 101. pages 232-236 (1990) disclosed various analogs of the C-terminal hexapeptide of ET-1, none of which were reported to be antagonists of ET-1.
Doherty, A. M., et al, Abstract, Second
International Conference on Endothelin, Tsukuba,
Japan, December 9, 1990, and the published manuscript (J. Cardiovasc. Pharm. 17 (Suppl. 7), 1991,
pp. 559-561) disclosed various analogs of the
C-terminal hexapeptide of ET-1, none of which
exhibited any functional activity. Copending United States Patent Application
Serial Number 07/995,480 discloses a series of novel antagonists of endothelin.
However, we have surprisingly and unexpectedly found that a series of C-terminal hexapeptide and related analogs of ET-1 are receptor antagonists of endothelin. Additional data for the activity of this series of peptides is found in the following
references (W.L. Cody, et al, J. Med. Chem., 1992, 35, 3301-3303., D.M. LaDouceur, et al, FASEB, 1992).
SUMMARY OF THE INVENTION
Accordingly, the present invention is a compound of Formula I
AA1-AA2 -AA3-AA4 -AA5-AA6 I wherein AA1 is
Figure imgf000013_0001
wherein R is hydrogen,
alkyl,
alkenyl,
alkynyl,
cycloalkyl,
cycloalkylalkyl,
aryl,
heteroaryl,
fluorenylmethyl, , wherein R2 and R3 are each the same or
Figure imgf000014_0001
different and each is
hydrogen,
alkyl,
alkenyl,
alkynyl,
cycloalkyl,
cycloalkylalkyl,
aryl,
arylalkyl,
heteroaryl, or
fluorenylmethyl,
Figure imgf000014_0002
, wherein R2 is as defined above,
-OR2, wherein R2 is as defined above, , wherein R2 and R3 are as defined above,
Figure imgf000014_0003
Figure imgf000014_0004
, wherein R9 is F, Cl, Br, or I,
-CH2-OR2, wherein R2 is as defined above, ,
Figure imgf000014_0005
wherein R2a is
defined above. ,
Figure imgf000014_0006
wherein R2a and R3 are as defined above excluding R3 is hydrogen, or
Figure imgf000014_0007
, wherein R2 is as defined above,
R1 is hydrogen or alkyl, Z is
-O-,
Figure imgf000015_0006
,
wherein m is zero or an integer of 1 or 2,
, wherein R2 is as defined above,
Figure imgf000015_0007
-(CH2)n-, wherein n is zero or an integer of 1, 2, 3, or 4,
-(CH2)n-CH=CH-(CH2)n-,
wherein n is as defined above,
Figure imgf000015_0005
,
, wherein R1 and R2 are as defined
Figure imgf000015_0004
above, or ,
Figure imgf000015_0003
wherein R2 and R3 are each the same or different and each is as defined above,
X and Y are the same and substituted at the same position on the aromatic ring and each may be one, two, three, or four substituents selected from the group consisting of hydrogen,
halogen,
alkyl,
-CO2R2, wherein R2 is as defined above,
, wherein R2 and R3 are as defined
Figure imgf000015_0002
above,
, wherein R2 and R3 are as defined
Figure imgf000015_0001
above, or nitro or
Figure imgf000016_0001
wherein R, Z, X, and Y are as defined above;
AA2 is
Figure imgf000016_0002
wherein R4 is
hydrogen,
alkyl,
alkenyl,
alkynyl,
cycloalkyl ,
aryl ,
heteroaryl,
,
Figure imgf000016_0003
wherein R2b and R3b are each the same or different and each is
hydrogen,
alkyl,
cycloalkyl,
aryl, or
heteroaryl,
-OR2b, wherein R2b is as defined above, ,
Figure imgf000016_0004
wherein R2b and R3b are each the same or different and each is as defined above for R2b and R3b,
Figure imgf000017_0005
, wherein R2b is as defined
above, wherein R2b is as defined
Figure imgf000017_0004
above, or
Figure imgf000017_0003
, wherein R2b is as defined
above, and
R1 and n are as defined above, or
AA2 is absent;
AA3 is
Figure imgf000017_0002
wherein R5 is
hydrogen,
alkyl,
aryl,
heteroaryl, ,
Figure imgf000017_0001
wherein R2b and R3b are each the same or
different and each is as defined above, , wherein R2b is as
Figure imgf000018_0001
defined above, or , wherein R2b is as
Figure imgf000018_0002
defined above, and
R1 and n are as defined above, or AA3 is absent;
AA4 and AA5 are each independently absent or each is independently
Figure imgf000018_0003
wherein R6 is hydrogen,
alkyl,
alkenyl,
alkynyl,
cycloalkyl,
aryl, or
heteroaryl, and
R1 and n are as defined above;
AA6 is
Figure imgf000018_0004
wherein R7 is
aryl or
heteroaryl,
R8 is , wherein R1 is as defined
Figure imgf000018_0005
above. -OR1, wherein R1 is as defined
above, , wherein R1 is as defined above, or
Figure imgf000019_0001
-CH2-OR1, wherein R1 is as defined above, and
R1 and n are as defined above;
*
stereochemistry at C in AA1, AA2, AA3, AA4, or AA5 is
D, L, or DL and
*
stereochemistry at C in AA6 is L; or a
pharmaceutically acceptable salt thereof.
Elevated levels of endothelin have been
postulated to be involved in a number of
pathophysiological states including diseases
associated with the cardiovascular system as well as various metabolic and endocrinological disorders. As antagonists of endothelin, the compounds of Formula I are useful in the treatment of hypertension,
myocardial infarction, metabolic, endocrinological and neurological disorders, congestive heart failure, endotoxic shock, subarachnoid hemorrhage, arrhythmias, asthma, and chronic and acute renal failure,
preeclampsia, atherosclerotic disorders including Raynaud's disease, restenosis, angina, cancer, pulmonary hypertension, ischemic disease, gastric mucosal damage, hemorrhagic shock, ischemic bowel disease, and diabetes.
A still further embodiment of the present
invention is a pharmaceutical composition for
administering an effective amount of a compound of
Formula I in unit dosage form in the treatment methods mentioned above.
Finally, the present invention is directed to methods for production of a compound of Formula I. DETAILED DESCRIPTION OF THE INVENTION
In the compounds of Formula I, the term "alkyl" means a straight or branched hydrocarbon radical having from 1 to 12 carbon atoms and includes, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, undecyl, dodecyl, and the like.
The term "alkenyl" means a straight or branched unsaturated hydrocarbon radical having from 2 to
12 carbon atoms and includes, for example, ethenyl, 2-propenyl, 1-butenyl, 2-butenyl, 1-pentenyl,
2-pentenyl, 3-methyl-3-butenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 3-heptenyl, 1-octenyl, 1-nonenyl,
1-decenyl, 1-undecenyl, 1-dodecenyl, and the like.
The term "alkynyl" means a straight or branched triple bonded unsaturated hydrocarbon radical having from 2 to 12 carbon atoms and includes, for example, ethynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 3-pentynyl, 1-hexynyl, 2-hexynyl,
3-hexynyl, 3-heptynyl, 1-octynyl, 2-octynyl,
1-nonynyl, 2-nonynyl, 3-nonynyl, 4-nonynyl, 1-decynyl, 2-decynyl, 2-undecynyl, 3-undecynyl, 3-dodecynyl, and the like.
The term "cycloalkyl" means a saturated
hydrocarbon ring which contains from 3 to 12 carbon atoms, for example, cyclopropyl, cyclobutyl,
cyclopentyl, cyclohexyl, adamantyl, and the like.
The term "cycloalkylalkyl" means a saturated hydrocarbon ring attached to an alkyl group wherein alkyl is as defined above. The saturated hydrocarbon ring contains from 3 to 12 carbon atoms. Examples of such are cyclopropylmethyl, cyclopentylmethyl, cyclohexylmethyl, adamantylmethyl and the like.
The terms "alkoxy" and "thioalkoxy" are O-alkyl or S-alkyl as defined above for alkyl. The term "aryl" means an aromatic radical which is a phenyl group, a benzyl group, a naphthyl group, a biphenyl group, a pyrenyl group, an anthracenyl group,
3,3-diphenylalanyl, 10,11-dihydro-5H-dibenzo[a,d]-(cyclohepten-5-yl)glycyl, or a fluorenyl group and the like, unsubstituted or substituted by 1 to
4 substituents selected from alkyl as defined above, alkoxy as defined above, thioalkoxy as defined above, hydroxy, thiol, nitro, halogen, amino,
Figure imgf000021_0002
wherein alkyl is as defined above,
Figure imgf000021_0001
wherein alkyl is as defined above, wherein alkyl is
Figure imgf000021_0003
as defined above, or aryl.
The term "arylalkyl" means an aromatic radical attached to an alkyl radical wherein aryl and alkyl are as defined above for example benzyl,
fluorenylmethyl and the like.
The term "heteroaryl" means a heteroaromatic radical which is 2-or 3-thienyl, 2- or 3-furanyl, 2-or 3-pyrrolyl, 2-, 4-, or 5-imidazolyl, 3-, 4-, or 5-pyrazolyl, 2-, 4-, or 5-thiazolyl, 3-, 4-, or
5-isothiazolyl, 2-, 4-, or 5-oxazolyl, 3-, 4-, or 5-isoxazolyl, 3- or 5-1,2,4-triazolyl, 4- or
5-1,2,3-triazolyl, tetrazolyl, 2-, 3-, or 4-pyridinyl, 3-, 4-, or 5-pyridazinyl, 2-pyrazinyl, 2-, 4-, or 5-pyrimidinyl, 2-, 3-, 4-, 5-, 6-, 7-, or
8-quinolinyl, 1-, 3-, 4-, 5-, 6-, 7-, or
8-isoquinolinyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-, 6-, or 7-benzo tb] thienyl, or 2-, 4-, 5-, 6-, or 7-benzoxazolyl, 2-, 4-, 5-, 6-,, or
7-benzimidazolyl, 2-, 4-, 5-, 6-, or 7-benzothiazolyl, unsubstituted or substituted by 1 to 2 substituents selected from alkyl as defined above, aryl as defined above, alkoxy as defined above, thioalkoxy as defined above, hydroxy, thiol, nitro, halogen, formyl, amino,
Figure imgf000022_0003
wherein alkyl is as defined above,
Figure imgf000022_0002
wherein alkyl is as defined above,
Figure imgf000022_0001
wherein alkyl is as defined above or phenyl.
The term "heterocycloalkyl" means 2- or
3-tetrahydrothieno, 2- or 3-tetrahydrofurano, 2- or 3-pyrrolidino, 2-, 4-, or 5-thiazolidino, 2-, 4-, or
5-oxazolidino, 2-, 3-, or 4-piperidino, N-morpholinyl or N-thiamorpholinyl.
"Halogen" is fluorine, chlorine, bromine or iodine.
The following table provides a list of
abbreviations and definitions thereof used in the present invention.
TABLE
Abbreviation* Amino Acid
Ala Alanine
Arg Arginine
Asn Asparagine
Asp Aspartic acid
Cys Cysteine
Glu Glutamic acid
Gln Glutamine
Gly Glycine
His Histidine
lle Isoleucine
Leu Leucine
Lys Lysine
Met Methionine
Phe Phenylalanine
Pro Proline
Ser Serine
Thr Threonine
Trp Tryptophan
Tyr Tyrosine
Val Valine
Abbreviation* Modified and Unusual Amino Acid
Bhg 10,ll-Dihydro-5H-dibenzo[a,d]- (cyclohepten-5-yl)glycine or
α-Amino-10,11-dihydro-5H-dibenzo- [a, d] cycloheptene-5-acetic acid
Bip (Paraphenyl) phenylalanine * If the optical activity of the amino acid is other than L(S), the amino acid or abbreviation is preceded by the appropriate configuration D(R) or DL(RS). Abbreviation* Modified and Unusual Amino Acid
(cont)
Dip 3,3-Diphenylalanine
3Hyp 3-Hydroxyproline
4Hyp 4-Hydroxyproline
N-MePhe N-Methylphenylalanine
N-MeAsp N-Methylaspartic acid
Nva Norvaline
Nle Norleucine
Orn Ornithine
Abu 2-Aminobutyric acid
Alg 2-Amino-4-pentenoic acid
(AllyIglycine)
Arg(NO2) NG-nitroarginine
Atm 2-Amino-3-(2-amino-5- thiazole)propanoic acid
Cpn 2-Amino-3-cyclopropanepropanoic acid
(Cyclopropylalanine)
Chx Cyclohexylalanine (Hexahydrophenylalanine)
Emg 2-Amino-4,5(RS)-epoxy-4-pentenoic
acid
His (Dnp) Nιm-2,4-Dinitrophenylhistidine
HomoGlu 2-Aminoadipic acid
HomoPhe 2-Amino-5-phenylpentanoic acid
(Homophenylalanine)
Met (O) Methionine sulfoxide
Met(O2) Methionine sulfone
1-Nal 3- (1'-Naphthyl) alanine
2-Nal 3- (2'-Naphthyl) alanine
Nia 2-Amino-3-cyanopropanoic acid
(Cyanoalanine)
Pgl Phenylglycine
Pgy 2-Aminopentanoic acid (Propylglycine) Abbreviation* Modified and Unusual Amino Acid
(cont)
Pha 2-Amino-6-(1-pyrrolo)-hexanoic acid Pyr 2-Amino-3-(3-pyridyl)-propanoic acid
(3-Pyridylalanine)
Tic 1,2,3,4-Tetrahydro-3- isoguinolinecarboxylic acid
Tza 2-Amino-3-(4-thiazolyl)-propanoic acid
Tyr(Ot-Bu) O-tertiary butyl -tyros ine
Tyr (OMe) O-Methyl-tyrosine
Tyr(OEt) O-Ethyl-tyrosine
Trp(For) Nin-Formyl-tryptophan
Bheg 5H-Dibenzo [a, d] cycloheptene glycine
Txg 9H-Thioxanthene glycine
Abbreviation Protecting Group
Ac Acetyl
Ada 1-Adamantyl acetic acid
Adoc Adamantyloxycarbonyl
Bzl Benzyl
MeBzl 4-Methylbenzyl
Z Benzyloxycarbonyl
2-Br-Z ortho-Bromobenzyloxycarbonyl
2-Cl-Z ortho-Chiorobenzyloxycarbonyl
Bom Benzyloxymethyl
Boc tertiary Butyloxycarbonyl
TBS tertiary Butyldimethylsilyl
Dnp 2,4-Dinitrophenyl
For Formyl
Fmoc 9-Fluorenylmethyloxycarbonyl
NO2 Nitro
Tos 4-Toluenesulfonyl (tosyl) Abbreviation Protecting Group (cont)
Trt Triphenylmethyl (trityl)
Ada 1-Adamantyl acetic acid
Bz Benzylcarbonyl
tBu t-Butylcarbonyl
CF3CO Trifluoroacetyl
Cxl Cyclohexylacetyl
Cxl (U) Cyclohexylurea
Et Propionyl
Pya 3-Pyridylacetyl
Me(U) Methylurea
Abbreviation Solvents and Reagents
HOAc Acetic acid
CH3CN Acetonitrile
DCM Dichloromethane
DCC N,N'-Dicyclohexylcarbodiimide
DIEA N,N-Diisopropylethylamine
DMF Dimethylformamide
HCl Hydrochloric acid
HF Hydrofluoric acid
HOBt 1-Hydroxybenzotriazole
KOH Potassium hydroxide
TFA Trifluoroacetic acid
MBHA Resin Methylbenzhydrylamine resin
PAM Resin 4-(Oxymethyl)-phenylacetamidomethyl resin
The compounds of Formula I are capable of further forming both pharmaceutically acceptable acid addition and/or base salts. All of these forms are within the scope of the present invention. Pharmaceutically acceptable acid addition salts of the compounds of Formula I include salts derived from nontoxic inorganic acids such as hydrochloric, nitric, phosphoric, sulfuric, hydrobromic, hydriodic, hydrofluoric, phosphorous, and the like, as well as the salts derived from nontoxic organic acids, such as aliphatic mono- and dicarboxylic acids,
phenyl-substituted alkanoic acids, hydroxy alkanoic acids, alkanedioic acids, aromatic acids, aliphatic and aromatic sulfonic acids, etc. Such salts thus include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, trifluoroacetate, propionate, caprylate, isobutyrate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, mandelate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, phthalate, benzenesulfonate,
toluenesulfonate, phenylacetate, citrate, lactate, maleate, tartrate, methanesulfonate, and the like.
Also contemplated are salts of amino acids such as arginate and the like and gluconate, galacturonate (see, for example, Berge, S. M., et al,
"Pharmaceutical Salts, " Journal of Pharmaceutical Science. 66. pp. 1-19 (1977)).
The acid addition salts of said basic compounds are prepared by contacting the free base form with a sufficient amount of the desired acid to produce the salt in the conventional manner. Preferably a peptide of Formula I can be converted to an acidic salt by treating with an aqueous solution of the desired acid, such that the resulting pH is less than 4. The solution can be passed through a C18 cartridge to absorb the peptide, washed with copious amounts of water, the peptide eluted with a polar organic solvent such as, for example, methanol, acetonitrile, aqueous mixtures thereof, and the like, and isolated by concentrating under reduced pressure followed by lyophilization. The free base form may be regenerated by contacting the salt form with a base and isolating the free base in the conventional manner. The free base forms differ from their respective salt forms somewhat in certain physical properties such as solubility in polar solvents, but otherwise the salts are equivalent to their respective free base for purposes of the present invention.
Pharmaceutically acceptable base addition salts are formed with metals or amines, such as alkali and alkaline earth metals or organic amines. Examples of metals used as cations are sodium, potassium,
magnesium, calcium, and the like. Examples of
suitable amines are N,N' -dibenzylethylenediamine, chloroprocaine, choline, diethanolamine,
dicyclohexylamine, ethylenediamine, N-methylglucamine, and procaine (see, for example, Berge, S. M., et al., "Pharmaceutical Salts," Journal of Pharmaceutical
Science, 66, pp. 1-19 (1977)).
The base addition salts of said acidic compounds are prepared by contacting the free acid form with a sufficient amount of the desired base to produce the salt in the conventional manner. Preferably, a peptide of Formula I can be converted to a base salt by treating with an aqueous solution of the desired base, such that the resulting pH is greater than 9. The solution can be passed through a C18 cartridge to absorb the peptide, washed with copious amounts of water, the peptide eluted with a polar organic solvent such as, for example, methanol, acetonitrile, aqueous mixtures thereof, and the like, and isolated by concentrating under reduced pressure followed by lyophilization. The free acid form may be regenerated by contacting the salt form with an acid and isolating the free acid in the conventional manner. The free acid forms differ from their respective salt forms somewhat in certain physical properties such as solubility in polar solvents, but otherwise the salts are equivalent to their respective free acid for purposes of the present invention.
Certain of the compounds of the present invention can exist in unsolvated forms as well as solvated forms, including hydrated forms. In general, the solvated forms, including hydrated forms, are
equivalent to unsolvated forms and are intended to be encompassed within the scope of the present invention.
Certain of the compounds of the present invention possess one or more chiral centers and each center may exist in the R(D) or S(L) configuration. The present invention includes all enantiomeric and epimeric forms as well as the appropriate mixtures thereof.
A preferred compound of Formula I is one wherein AA1 is
Figure imgf000029_0001
wherein R is ,
Figure imgf000029_0002
wherein R2 and R3 are each the same or different and each is
hydrogen,
alkyl,
alkenyl,
alkynyl,
cycloalkyl,
cycloalkylalkyl,
aryl,
arylalkyl,
heteroaryl, or
fluorenylmethyl, , wherein R2 and R3 are as
Figure imgf000030_0010
defined above.
Figure imgf000030_0009
, wherein R9 is F, Cl, Br, or I,
Figure imgf000030_0008
, wherei.n R3 is as defined
above, or
Figure imgf000030_0007
,
wherein R3 is as defined above excluding R3 is hydrogen,
Z is -O-,
Figure imgf000030_0006
,
wherein m is zero or an integer of 1 or 2,
, wherein R2 is as defined above,
Figure imgf000030_0005
Figure imgf000030_0004
, wherein n is zero or an integer of
1, 2, 3, or 4,
- (CH2)n-CH=CH-(CH2)n-, wherein n is as defined above,
Figure imgf000030_0003
,
, wherein R1 is hydrogen or alkyl,
Figure imgf000030_0002
,
Figure imgf000030_0001
wherein R2 and R3 are each the same or different and each is as defined above and X and Y are the same and substituted at the same position on the aromatic ring and each substituent is selected from the group consisting of
hydrogen,
halogen, or
alkyl;
AA^ is
Figure imgf000031_0001
wherein R4 is hydrogen,
alkyl,
alkenyl,
alkynyl,
cycloalkyl,
aryl,
heteroaryl,
,
Figure imgf000031_0002
wherein R2b and R3b are each the same or different and each is
hydrogen,
alkyl,
cycloalkyl,
aryl, or
heteroaryl,
-OR2b, wherein R2b is as
defined above,
.
,
Figure imgf000031_0003
wherein R2b and R3b are each the same or different and each is as defined above for
R2b and R3b,
Figure imgf000032_0006
, wherein R2b is as defined above,
Figure imgf000032_0005
, wherein R2b is as
defined above, or
Figure imgf000032_0004
, wherein R2b is as
defined above, and n is as defined above or
AA2 is absent :
AA3 is
Figure imgf000032_0003
wherein R5 is aryl,
heteroaryl, ,
Figure imgf000032_0002
wherein R2b and R3b are each the same or different and each is as defined above,
Figure imgf000032_0001
, wherein R2b is as defined
above, or , wherein R2b is as defined
Figure imgf000033_0004
above, and
n is as defined above, or
AA3 is absent;
AA4 and AA5 are each independently absent or each is
Figure imgf000033_0003
wherein R6 is hydrogen
alkyl,
alkenyl,
alkynyl,
cycloalkyl,
aryl, or
heteroaryl, and
n is as defined above;
AA6 is
Figure imgf000033_0002
wherein R7 is aryl or
heteroaryl, and
n is as defined above, or
wherein R7, R1, and r
defined above,
Figure imgf000033_0001
*
stereochemistry at CH m AA 1, AA2 , AA3 , AA4, or AA5 is
D,L, or DL, and
*
stereochemistry at CH in AA6 is L; or a
pharmaceutically acceptable salt thereof.
A more preferred compound of Formula I is one wherein AA1 is
Figure imgf000034_0001
wherein R is ,
Figure imgf000034_0002
wherein R2 and R3 are each the same or different and each is
hydrogen,
alkyl,
aryl, or
fluorenylmethyl, , wherein R2 and R3 are
as defined above.
Figure imgf000034_0004
,wherein R9 is F, Cl, Br, or I,
Figure imgf000034_0005
or , wherein R10 is hydrogen,
Figure imgf000034_0003
alkyl, aryl, or
arylalkyl, excluding R10 is hydrogen,
Z is -O-,
-S-,
-NH-, -(CH2)n, wherein n is zero or an integer
of 1, 2, 3, or 4, or
Figure imgf000035_0004
,
wherein n is zero or an integer of
1 or 2 and
X and Y are each the same and substituted at the same position on the aromatic ring and each substituent is selected from the group consisting of
hydrogen,
halogen, or
alkyl;
,
Figure imgf000035_0003
wherein R4 is hydrogen,
alkyl,
aryl,
heteroaryl,
,
Figure imgf000035_0002
wherein R2b and R3b are each the same or
different and each is hydrogen or alkyl, '
Figure imgf000035_0001
wherein R2b and R3b are each the same or
different and each is hydrogen or alkyl,
Figure imgf000036_0001
wherein R2b is as defined above, or , wherein R2b is as
Figure imgf000036_0002
defined above, and n is an integer of 1, 2, 3, or 4 or
AA2 is absent;
AA3 is , wherein R3 is
Figure imgf000036_0003
aryl,
heteroaryl,
Figure imgf000036_0004
, wherein R3b is
hydrogen or
alkyl.
Figure imgf000036_0005
, wherein R2b is
hydrogen or
alkyl, or
Figure imgf000036_0006
, wherein R2b is
hydrogen or
alkyl, and
n is an integer of 1, 2, 3, or 4;
AA4 and AA5 are each independently
Figure imgf000036_0007
wherein R6 is hydrogen,
alkyl,
cycloalkyl, or
aryl, and
n is an integer of 1, 2, 3, or 4;
AA6 is
Figure imgf000037_0001
wherein R7 is aryl or heteroaryl, and
n is zero or an integer of 1, 2, 3, or 4, or
wherein R7, R1, and n are as
defined above,
Figure imgf000037_0002
stereochemistry at CH m AA1, AA2, AA3, AA4, or AA5 i.s D, L, or DL and
*
stereochemistry at CH in AA6 is L; or a
pharmaceutically acceptable salt thereof.
Particularly valuable are:
L-Bhg-Leu-Asp-Ile-Ile-Trp;
D-Bhg-Leu-Asp-Ile-Ile-Trp;
Ac-L-Bhg-Leu-Asp-Ile-Ile-Trp;
Ac-D-Bhg-Leu-Asp-Ile-Ile-Trp;
Ac-D-Bhg-Orn-Asp-Ile-Ile-Trp;
Ac-D-Bhg-Lys-Asp-Ile-Ile-Trp;
Ac-D-Bhg-Asp-Asp-Ile-Ile-Trp;
Ac-D-Bhg-Glu-Asp-Ile-Ile-Trp;
Ac-D-Bhg-Phe-Asp-Ile-Ile-Trp;
Ac-D-Bhg-Arg-Asp-Ile-Ile-Trp;
Ac-D-Bhg-Asp-Ile-Ile-Trp;
Fmoc-D-Bhg-Leu-Asp-Ile-Ile-Trp;
Fmoc-D-Bhg-Orn-Asp-Ile-Ile-Trp; Fmoc-D-Bhg-Lys-Asp-Ile-Ile-Trp;
Fmoc-D-Bhg-Asp-Asp-Ile-Ile-Trp;
Fmoc-D-Bhg-Glu-Asp-Ile-Ile-Trp;
Emoc-D-Bhg-Phe-Asp-Ile-Ile-Trp; Fmoc-D-Bhg-Arg-Asp-Ile-Ile-Trp;
Fmoc-D-Bhg-Asp-Ile-Ile-Trp;
Ac-D-Bhg-Leu-Phe-Ile-Ile-Trp;
Ac-D-Bhg-Leu-Asn-Ile-Ile-Trp;
Ac-D-Bhg-Leu-Glu-Ile-Ile-Trp; Ac-D-Bhg-Leu-Gln-Ile-Ile-Trp;
Ac-D-Bhg-Leu-Tyr-Ile-Ile-Trp;
Ac-D-Bhg-Leu-1-Nal-Ile-Ile-Trp;
Ac-D-Bhg-Leu-2-Nal-Ile-Ile-Trp;
Ac-D-Bhg-Leu-Trp-Ile-Ile-Trp; Ac-D-Bhg-Leu-Asp-Val-Ile-Trp;
Ac-D-Bhg-Leu-Asp-Ile-Val-Trp;
Ac-D-Bhg-Leu-Asp-Chx-Ile-Trp;
Ac-D-Bhg-Leu-Asp-Ile-Chx-Trp;
Ac-D-Bhg-Arg-Asp-Ile-Chx-Trp; Ac-D-Bhg-Lys-Asp-Ile-Chx-Trp;
Ac-D-Bhg-Orn-Asp-Ile-Chx-Trp;
Ac-D-Bhg-Asp-Asp-Ile-Chx-Trp;
Ac-D-Bhg-Glu-Asp-Ile-Chx-Trp;
Fmoc-D-Bhg-Leu-Phe-Ile-Ile-Trp; Emoc-D-Bhg-Leu-Asn-Ile-Ile-Trp;
Fmoc-D-Bhg-Leu-Glu-Ile-Ile-Trp;
Fmoc-D-Bhg-Leu-Gln-Ile-Ile-Trp;
Fmoc-D-Bhg-Leu-Tyr-Ile-Ile-Trp;
Fmoc-D-Bhg-Leu-Asp-Val-Ile-Trp; Fmoc-D-Bhg-Leu-Ass-Ile-Val-Trp;
Fmoc-D-Bhg-Leu-Asp-Chx-Ile-Trp;
Fmoc-D-Bhg-Arg-Asp-Chx-Ile-Trp;
Fmoc-D-Bhg-Lys-Asp-Chx-Ile-Trp;
Fmoc-D-Bhg-Orn-Asp-Chx-Ile-Trp; Fmoc-D-Bhg-Asp-Asp-Chx-Ile-Trp;
Fmoc-D-Bhg-Glu-Asp-Chx-Ile-Trp;
Fmoc-D-Bhg-Leu-Asp-Ile-Chx-Trp; Fmoc-D-Bhg-Arg-Asp-Ile-Chx-Trp;
Fmoc-D-Bhg-Lys-Asp-Ile-Chx-Trp;
Fmoc-D-Bhg-Orn-Asp-Ile-Chx-Trp;
Fmoc-D-Bhg-Asp-Asp-Ile-Chx-Trp; Fmoc-D-Bhg-Glu-Asp-Ile-Chx-Trp;
Ac-D-Bheg-Leu-Asp-Ile-Ile-Trp;
Ac-D-Bheg-Orn-Asp-Ile-Ile-Trp;
Ac-D-Bheg-Lys-Asp-Ile-Ile-Trp;
Ac-D-Bheg-Asp-Asp-Ile-Ile-Trp; Ac-D-Bheg-Glu-Asp-Ile-Ile-Trp;
Ac-D-Bheg-Phe-Asp-Ile-Ile-Trp;
Ac-D-Bheg-Arg-Asp-Ile-Ile-Trp;
Ac-D-Bheg-Asp-Ile-Ile-Trp;
Fmoc-D-Bheg-Leu-Asp-Ile-Ile-Trp; Fmoc-D-Bheg-Orn-Asp-Ile-Ile-Trp;
Fmoc-D-Bheg-Lys-Asp-Ile-Ile-Trp;
Fmoc-D-Bheg-Asp-Asp-Ile-Ile-Trp;
Fmoc-D-Bheg-Glu-Asp-Ile-Ile-Trp;
Fmoc-D-Bheg-Phe-Asp-Ile-Ile-Trp; Fmoc-D-Bheg-Arg-Asp-Ile-Ile-Trp;
Fmoc-D-Bheg-Asp-Ile-Ile-Trp;
Ac-D-Bheg-Leu-Phe-Ile-Ile-Trp;
Ac-D-Bheg-Leu-Asn-Ile-Ile-Trp;
Ac-D-Bheg-Leu-Glu-Ile-Ile-Trp; Ac-D-Bheg-Leu-Gln-Ile-Ile-Trp;
Ac-D-Bheg-Leu-Tyr-Ile-Ile-Trp;
Ac-D-Bheg-Leu-1-Nal-Ile-Ile-Trp;
Ac-D-Bheg-Leu-2-Nal-Ile-Ile-Trp;
Ac-D-Bheg-Leu-Trp-Ile-Ile-Trp; Ac-D-Bheg-Leu-Asp-Val-Ile-Trp;
Ac-D-Bheg-Leu-Asp-Ile-Val-Trp;
Ac-D-Bheg-Leu-Asp-Chx-Ile-Trp;
Ac-D-Bheg-Leu-Asp-Ile-Chx-Trp;
Ac-D-Bheg-Arg-Asp-Ile-Chx-Trp; Ac-D-Bheg-Lys-Asp-Ile-Chx-Trp;
Ac-D-Bheg-Orn-Asp-Ile-Chx-Trp;
Ac-D-Bheg-Asp-Asp-Ile-Chx-Trp; Ac-D-Bheg-Glu-Asp-Ile-Chx-Trp; Fmoc-D-Bheg-Leu-Phe-Ile-Ile-Trp Emoc-D-Bheg-Leu-Asn-Ile-Ile-Trp Fmoc-D-Bheg-Leu-Glu-Ile-Ile-Trp Fmoc-D-Bheg-Leu-Gln-Ile-Ile-Trp Fmoc-D-Bheg-Leu-Tyr-Ile-Ile-Trp Fmoc-D-Bheg-Leu-Asp-Val-Ile-Trp Fmoc-D-Bheg-Leu-Asp-Ile-Val-Trp Fmoc-D-Bheg-Leu-Asp-Chx-Ile-Trp Fmoc-D-Bheg-Arg-Asp-Chx-Ile-Trp Fmoc-D-Bheg-Lys-Asp-Chx-Ile-Trp Fmoc-D-Bheg-Orn-Asp-Chx-Ile-Trp Fmoc-D-Bheg-Asp-Asp-Chx-Ile-Trp Fmoc-D-Bheg-Glu-Asp-Chx-Ile-Trp Fmoc-D-Bheg-Leu-Asp-Ile-Chx-Trp Fmoc-D-Bheg-Arg-Asp-Ile-Chx-Trp Emoc-D-Bheg-Lys-Asp-Ile-Chx-Trp Fmoc-D-Bheg-Orn-Asp-Ile-Chx-Trp Fmoc-D-Bheg-Asp-Asp-Ile-Chx-Trp Fmoc-D-Bheg-Glu-Asp-Ile-Chx-Trp Ac-D-Txg-Leu-Asp-Ile-Ile-Trp; Ac-D-Txg-Orn-Asp-Ile-Ile-Trp; Ac-D-Txg-Lys-Asp-Ile-Ile-Trp; Ac-D-Txg-Asp-Asp-Ile-Ile-Trp; Ac-D-Txg-Glu-Asp-Ile-Ile-Trp; Ac-D-Txg-Phe-Asp-Ile-Ile-Trp; Ac-D-Txg-Arg-Asp-Ile-Ile-Trp; Ac-D-Txg-Asp-Ile-Ile-Trp;
Fmoc-D-Txg-Leu-Asp-Ile-Ile-Trp; Fmoc-D-Txg-Orn-Asp-Ile-Ile-Trp; Fmoc-D-Txg-Lys-Asp-Ile-Ile-Trp; Fmoc-D-Txg-Asp-Asp-Ile-Ile-Trp; Fmoc-D-Txg-Glu-Asp-Ile-Ile-Trp; Fmoc-D-Txg-Phe-Asp-Ile-Ile-Trp; Fmoc-D-Txg-Arg-Asp-Ile-Ile-Trp; Fmoc-D-Txg-Asp-Ile-Ile-Trp; Ac-D-Txg-Leu-Phe-Ile-Ile-Trp; Ac-D-Txg-Leu-Asn-Ile-Ile-Trp; Ac-D-Txg-Leu-Glu-Ile-Ile-Trp; Ac-D-Txg-Leu-Gln-Ile-Ile-Trp; Ac-D-Txg-Leu-Tyr-Ile-Ile-Trp; Ac-D-Txg-Leu-1-Nal-Ile-Ile-Trp; Ac-D-Txg-Leu-2-Nal-Ile-Ile-Trp; Ac-D-Txg-Leu-Trp-Ile-Ile-Trp; Ac-D-Txg-Leu-Asp-Val-Ile-Trp; Ac-D-Txg-Leu-Asp-Ile-Val-Trp; Ac-D-Txg-Leu-Asp-Chx-Ile-Trp; Ac-D-Txg-Leu-Asp-Ile-Chx-Trp; Ac-D-Txg-Arg-Asp-Ile-Chx-Trp; Ac-D-Txg-Lys-Asp-Ile-Chx-Trp; Ac-D-Txg-Orn-Asp-Ile-Chx-Trp; Ac-D-Txg-Asp-Asp-Ile-Chx-Trp; Ac-D-Txg-Glu-Asp-Ile-Chx-Trp; Fmoc-D-Txg-Leu-Phe-Ile-Ile-Trp Fmoc-D-Txg-Leu-Asn-Ile-Ile-Trp Fmoc-D-Txg-Leu-Glu-Ile-Ile-Trp Fmoc-D-Txg-Leu-Gln-Ile-Ile-Trp Fmoc-D-Txg-Leu-Tyr-Ile-Ile-Trp Fmoc-D-Txg-Leu-Asp-Val-Ile-Trp Fmoc-D-Txg-Leu-Asp-Ile-Val-Trp Fmoc-D-Txg-Leu-Asp-Chx-Ile-Trp Fmoc-D-Txg-Arg-Asp-Chx-Ile-Trp Fmoc-D-Txg-Lys-Asp-Chx-Ile-Trp Fmoc-D-Txg-Orn-Asp-Chx-Ile-Trp Fmoc-D-Txg-Asp-Asp-Chx-Ile-Trp Fmoc-D-Txg-Glu-Asp-Chx-Ile-Trp Fmoc-D-Txg-Leu-Asp-Ile-Chx-Trp Fmoc-D-Txg-Arg-Asp-Ile-Chx-Trp Fmoc-D-Txg-Lys-Asp-Ile-Chx-Trp Fmoc-D-Txg-Orn-Asp-Ile-Chx-Trp Fmoc-D-Txg-Asp-Asp-Ile-Chx-Trp Fmoc-D-Txg-Glu-Asp-Ile-Chx-Trp Et-D-Bhg-Leu-Asp-Ile-Ile-Trp; Bz-D-Bhg-Leu-Asp-Ile-Ile-Trp; Pya-D-Bhg-Leu-Asp-Ile-Ile-Trp;
Cxl-D-Bhg-Leu-Asp-Ile-Ile-Trp;
Ada-D-Bhg-Leu-Asp-Ile-Ile-Trp;
Cxl(U)-D-Bhg-Leu-Asp-Ile-Ile-Trp;
Me(U)-D-Bhg-Leu-Asp-Ile-Ile-Trp;
tBu-D-Bhg-Leu-Asp-Ile-Ile-Trp;
CF3CO-D-Bhg-Leu-Asp-Ile-Ile-Trp;
Et-D-Bheg-Leu-Asp-Ile-Ile-Trp;
Bz-D-Bheg-Leu-Asp-Ile-Ile-Trp;
Pya-D-Bheg-Leu-Asp-Ile-Ile-Trp;
Cxl-D-Bheg-Leu-Asp-Ile-Ile-Trp;
Ada-D-Bheg-Leu-Asp-Ile-Ile-Trp;
Cxl(U)-D-Bheg-Leu-Asp-Ile-Ile-Trp;
Me(U)-D-Bheg-Leu-Asp-Ile-Ile-Trp;
tBu-D-Bheg-Leu-Asp-Ile-Ile-Trp;
CF3CO-D-Bheg-Leu-Asp-Ile-Ile-Trp;
Ac-D-Bhg-Leu-Asp-Phe-Ile-Trp;
Ac-D-Bhg-Orn-Asp-Phe-Ile-Trp;
Ac-D-Bhg-Lys-Asp-Phe-Ile-Trp;
Ac-D-Bhg-Asp-Asp-Phe-Ile-Trp;
Ac-D-Bhg-Glu-Asp-Phe-Ile-Trp;
Ac-D-Bhg-Phe-Asp-Phe-Ile-Trp;
Ac-D-Bhg-Arg-Asp-Phe-Ile-Trp;
Ac-D-Bheg-Leu-Asp-Phe-Ile-Trp;
Ac-D-Bheg-Orn-Asp-Phe-Ile-Trp;
Ac-D-Bheg-Lys-Asp-Phe-Ile-Trp;
Ac-D-Bheg-Asp-Asp-Phe-Ile-Trp;
Ac-D-Bheg-Glu-Asp-Phe-Ile-Trp;
Ac-D-Bheg-Phe-Asp-Phe-Ile-Trp; and
Ac-D-Bheg-Arg-Asp-Phe-Ile-Trp;
or a pharmaceutically acceptable acid or base addition salt thereof.
The compounds of Formula I are valuable
antagonists of endothelin. The tests employed
indicate that compounds of Formula I possess
endothelin antagonist activity. Thus, the compounds of Formula I were tested for their ability to inhibit [125I] -ET-1 ( [125I] -Endothelin-1) binding in a receptor assay according to the following procedures:
ENDOTHELIN RECEPTOR BINDING ASSAY-A (ERBA-A)
INTACT CELL BINDING OF [125I] -ET-1
Materials and Terms Used:
Cells
The cells used were rabbit renal artery vascular smooth muscle cells grown in a 48-well dish (1 cm2) (confluent cells).
Growth Media
The growth media was Dulbecco's Modified
Eagles/Ham's F12 which contained 10% fetal bovine serum and antibiotics (penicillin/streptomycin/ fungizone).
Assay Buffer
The assay buffer was a medium 199 containing
Hanks salts and 25 mM Hepes buffer (Gibco 380-2350AJ), supplemented with penicillin/streptomycin/fungizone (0.5%) and bovine serum albumin (1 mg/mL). [125I] -ET-1
Amersham radioiodinated endothelin-1 [125I]-ET-1 was used at final concentration of 20,000 cpm/0.25 mL (25 pM). Protocol
First, add 0.5 mL warm assay buffer (described above) to the aspirated growth media and preincubate for 2 to 3 hours in a 37°C water bath (do not put back in the 5% carbon dioxide). Second, remove the assay buffers, place the dish on ice, and add 150 μL of cold assay buffer described above to each well. Third, add 50 mL each of cold [125I] -ET-1 and competing ligand to the solution (at the same time if possible). Next, place dish in a 37°C water bath for about 2 hours and gently agitate the dish every 15 minutes. Discard the radioactive incubation mixture in the sink and wash wells 3 times with 1 mL of cold phosphate buffered saline. Last, add 250 mL of 0.25 molar sodium
hydroxide, agitate for 1 hour on a rotator, and then transfer the sodium hydroxide extract to gamma
counting tubes and count the radioactivity.
ENDOTHELIN RECEPTOR BINDING ASSAY-B (ERBA-B)
[125I] -ET-1 BINDING IN RAT CEREBELLAR MEMBRANES
Materials and Terms Used:
Tissue Buffer
The tissue is made up of 20 mM tris (hydroxy-methyl)aminomethane hydrochloride (Trizma) buffer, 2 mM ethylenediaminetetra acetate, 100 μM
phenylmethylsulfonyl fluoride.
Tissue Preparation
First, thaw one aliquot of frozen rat cerebellar membranes (2 mg protein in 0.5 mL) . Next, add 0.5 mL membrane aliquot to 4.5 mL cold tissue buffer, polytron at 7,500 revolutions per minute for
10 seconds. Finally, dilute tissue suspension 1/100 (0.1 mL suspension + 9.9 mL tissue buffer), polytron again, and place ice. Dilution Buffer
Medium 199 with Hank's salts plus 25 mM Hepes + 1 mg/mL bovine serum albumin.
[125I]-ET-1
Amersham [125I] -ET-1 (aliquots of 2 × 106 cpm per
100 mL aliquot of [125I] -ET- 1 with 5.2 mL dilution buffer, place on ice until use (final concentration will be 20,000 cpm per tube, or 25 pM).
Protocol
Add 50 μL each of cold [125] -ET-1 and competing ligand to tubes on ice. Mix in 150 μL of tissue to each tube, vortex briefly, then tap to force all liquids to bottom (total assay volume = 250 μL) . Then place the tubes in a 37°C water bath for 2 hours.
Add 2.5 mL cold wash buffer (50 mM Trizma buffer) to each tube, filter, and then wash tube with
additional 2.5 mL wash buffer and add to filter.
Finally, wash filters with an additional 2.5 mL of cold wash buffer.
Count filters for radioactivity in gamma counter.
IN VITRO INHIBITION OF ET-1 STIMULATED ARACHIDONIC ACID RELEASE (AAR) IN CULTURED RABBIT VASCULAR SMOOTH
MUSCLE CELLS BY COMPOUNDS OF FORMULA I
Antagonist activity is measured by the ability of added compounds to reduce endothelin-stimulated arachidonic acid release in cultured vascular smooth muscle cells as arachidonic acid release (AAR).
[3H] Arachidonic Acid Loading Media (LM) is
DME/F12 + 0.5% FCS × 0.25 mCi/mL [3H] arachidonic acid (Amersham). Confluent monolayers of cultured rabbit renal artery vascular smooth muscle cells were
incubated in 0.5 mL of the LM over 18 hours, at 37°C, in 5% CO2. The LM was aspirated and the cells were washed once with the assay buffer (Hank's BSS + 10 mM HEPES + fatty acid-free BSA (1 mg/mL)), and incubated for 5 minutes with 1 mL of the prewarmed assay buffer. This solution was aspirated, followed by an additional 1 mL of prewarmed assay buffer, and further incubated for another 5 minutes. A final 5-minute incubation was carried out in a similar manner. The same
procedure was repeated with the inclusion of 10 μL of the test compound (1 nM to 1 μM) and 10 μL ET-1
(0.3 nM) and the incubation was extended for
30 minutes. This solution was then collected, 10 μL of scintillation cocktail was added, and the amount of [3H] arachidonic acid was determined in a liquid scintillation counter. IN VITRO ANTAGONISM OF ET-1 STIMULATED
VASOCONSTRICTION IN THE RABBIT FEMORAL ARTERY (ETA)
AND SARAFOTOXIN 6c
STIMULATED VASOCONSTRICTION IN THE RABBIT PULMONARY
ARTERY (ETB)
Male New Zealand rabbits were killed by cervical dislocation and exsanguination. Femoral and pulmonary arteries were isolated, cleaned of connective tissue, and cut into 4-mm rings. The endothelium was denuded by placing the rings over hypodermic tubing (32 guage for femoral rings and 28 guage for pulmonary rings, Small Parts, Ine, Miami, Florida) and gently rolling them. Denuded rings were mounted in 20 mL organ baths containing Krebs-bicarbonate buffer (composition in mM: NaCl, 118.2; NaHCO3, 24.8; KCl, 4.6; MgSO4
7·H2O, 1.2; KH2PO4, 1.2; CaCl2·2H2O; Ca-Na2 EDTA, 0.026; dextrose, 10.0), that was maintained at 37°C and gassed continuously with 5% CO2 in oxygen
(pH 7.4). Resting tension was adjusted to 3.0 g for femoral and 4.0 g pulmonary arteries; the rings were left for 90 minutes to equilibrate. Vascular rings were tested for lack of functional endothelium (i.e., lack of an endothelium-dependent relaxation response to carbachol (1.0 μM.) in norepinephrine (0.03 μM) contracted rings. Agonist peptides, ET-1 (femoral), and S6c (pulmonary), were cumulatively added at
10-minute intervals. The ET antagonists were added 30 minutes prior to adding the agonist and pA 2 values were calculated (Table I).
The data in Table I below show the endothelin receptor binding and antagonist activity of
representative compounds of Formula I.
TABLE I. Biological Activity of Compounds of Formula I
Example ERBA-A ERBA-B AAR
Compound pA 2
Number IC50 (μm) IC50 (/XM) IC50 (μM) Femoral Pulmonary
1 Ac-D-Bhg-Leu-Asp-Ile-Ile-Trp 0.0026 0.0X9 0.0049
2 D-Bhg-Leu-Asp-Ile-Ile-Trp 0.45 2.1 0.10
3 L-Bhg-Leu-Asp-Ile-Ile-Trp 2.5 3.0 2.36
4 Ac-L-Bhg-Leu-Asp-Ile-Ile-Trp 0.56 0.71 0.56
5 Ac-D-Txg-Leu-Asp-Ile-Ile-Trp 0.018 0.18
6 Ac-D-Bheg-Leu-Asp-Ile-Ile-Trp 0.005 0.019 0.003 6.8 7.4
7 Ac-D-Bhg-Om-Asp-Ile-Ile-Trp 0.022 1.5 0.037 6.5
8 Ac-D-Bhg-Glu-Asp-Ile-Ile-Trp 0.0055 0.022 0.007 6.5 7.0
9 Ac-D-Bhg-Leu-Asp-Ile-Ile-Trp, 2Na+ 0.004 0.015 0.0049 6.9 7.1
General Method for Preparing Compounds of Formula I
The compounds of Formula I may be prepared by solid phase peptide synthesis on a peptide
synthesizer, for example, an Applied Biosystems 430A peptide synthesizer using activated esters or
anhydrides of N-alpha-Boc protected amino acids, on PAM or MBHA resins. Additionally, the compounds of Formula I may also be prepared by conventional solution peptide synthesis. Amino acid side chains are protected as follows: BzKAsp, Glu, Ser),
2-Cl-Z(Lys), 2-Br-Z(Tyr), Bom(His), For(Trp), and MeBzl(Cys). Each peptide resin (1.0 g) is cleaved with 9 mL of HF and 1 mL of anisole or p-cresol as a scavenger (60 minutes, 0°C). The peptide resin is washed with cyclohexane, extracted with 30% aqueous HOAc, followed by glacial HOAc, concentrated under reduced pressure, and lyophilized. (A peptide containing For (Trp) is dissolved in 0°C, the pH is adjusted to 12.5 with IN KOH (2 minutes), neutralized with glacial HOAc, desalted on C18 (as described below), and lyophilized. The crude peptide is purified by preparative reversed phase high
performance liquid chromatography (RP-HPLC) on a C18 column (2.2 × 25.0 cm, 15.0 mL/min) with a linear gradient of 0.1% TFA in water to 0.1% TFA in
acetonitrile and lyophilized. The homogeneity and composition of the resulting peptide is verified by RP-HPLC, capillary electrophoresis, thin layer chromatography (TLC), proton nuclear magnetic
resonance spectrometry (NMR), and fast atom
bombardment mass spectrometry (FAB-MS).
The compounds of the present invention can be prepared and administered in a wide variety of oral and parenteral dosage forms. Thus, the compounds of the present invention can be administered by
injection, that is, intravenously, intramuscularly, intracutaneously, sύbcutaneously, mtraduodenally, or intraperitoneally. Also, the compounds of the present invention can be administered by inhalation, for example, intranasally. Additionally, the compounds of the present invention can be administered
transdermally. It will be obvious to those skilled in the art that the following dosage forms may comprise as the active component, either a compound of
Formula I or a corresponding pharmaceutically
acceptable salt of a compound of Formula I.
For preparing pharmaceutical compositions from the compounds of the present invention,
pharmaceutically acceptable carriers can be either solid or liquid. Solid form preparations include powders, tablets, pills, capsules, cachets,
suppositories, and dispersible granules. A solid carrier can be one or more substances which may also act as diluents, flavoring agents, binders,
preservatives, tablet disintegrating agents, or 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.
The powders and tablets preferably contain from five or ten to about seventy percent of the active compound. Suitable carriers are magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, a low melting wax, cocoa butter, and the like. The term "preparation" is intended to include the formulation of the active compound 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 and lozenges are included. Tablets, powders, capsules, pills, cachets, and lozenges can be used as solid dosage forms suitable for oral administration.
For preparing suppositories, a low melting wax, such as a mixture of fatty acid glycerides or cocoa butter, is first melted and the active component is dispersed homogeneously therein, as by stirring. The molten homogenous mixture is then poured into
convenient sized molds, allowed to cool, and thereby to solidify.
Liquid form preparations include solutions, suspensions, and emulsions, for example, water or water propylene glycol solutions. For parenteral injection liquid preparations can be formulated in solution in aqueous polyethylene glycol solution.
Aqueous solutions suitable for oral use can be prepared by dissolving the active component in water and adding suitable colorants, flavors, stabilizing and thickening agents as desired.
Aqueous suspensions suitable for oral use can be made by dispersing the finely divided active component in water with viscous material, such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, and other well-known
suspending agents.
Also included are solid form preparations which are intended to be converted, shortly before use, to liquid form preparations for oral administration.
Such liquid forms include solutions, suspensions, and emulsions. These preparations may contain, in
addition to the active component, colorants, flavors, stabilizers, buffers, artificial and natural
sweeteners, dispersants, thickeners, solubilizing agents, and the like.
The pharmaceutical preparation is preferably in unit dosage form. In such form the preparation is subdivided 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 preparation, such as packeted tablets, capsules, and powders in vials or ampoules. Also, the unit dosage form can be a
capsules, tablet, cachet, or lozenge itself, or it can be the appropriate number of any of these in packaged form.
The quantity of active component in a unit dose preparation may be varied or adjusted from 0.1 mg to 100 mg preferably 0.5 mg to 100 mg according to the particular application and the potency of the active component. The composition can, if desired, also contain other compatible therapeutic agents.
In therapeutic use as antagonist of endothelin, the compounds utilized in the pharmaceutical method of this invention are administered at the initial dosage of about 0.01 mg to about 20 mg per kilogram daily. A daily dose range of about 0.01 mg to about 10 mg per kilogram is preferred. The dosages, however, may be varied depending upon the requirements of the patient, the severity of the condition being treated, and the compound being employed. Determination of the proper dosage for a particular situation is within the skill of the art. Generally, treatment is initiated with smaller dosages which are less than the optimum dose of the compound. Thereafter, the dosage is increased by small increments until the optimum effect under the circumstances is reached. For convenience, the total daily dosage may be divided and administered in portions during the day, if desired.
The following nonlimiting examples illustrate the inventors' preferred methods for preparing the
compounds of the invention. EXAMPLE 1
Ac-D-Bhg-Leu-Asp-Ile-Ile-Trp
The linear hexapeptide is prepared by standard solid phase synthetic peptide methodology utilizing a Boc/benzyl strategy (Stewart, J. M. and Young, J. D., Solid Phase Peptide Synthesis. Pierce Chemical Co., Rockford, IL, 1984). All protected amino acids and reagents are obtained from commercial sources with the exception of N-α-Boc-DL-Bhg and are not further purified. The protected peptide resin is prepared on an Applied Biosystems 430A Peptide Synthesizer, utilizing protocols supplied for a dicyclohexyl-carbodiimide-mediated coupling scheme (Standard 1.0, Version 1.40). Starting with 0.710 g of
N-α-Boc-Trp-PAM resin (0.70 meq/g, 0.497 meq of
Boc-Trp(For) total) the protected peptide is prepared by the stepwise coupling of the following amino acids (in order of addition): N-α-Boc-Ile.0.5H2O,
N-Q!-Boc-Ile.0.5H2O, N-α-Boc-Asp(Bzl), N-α-Boc-Leu-H2O, and N-α-Boc-DL-Bhg. A typical cycle for the coupling of an individual amino acid residue is illustrated below (reproduced from the ABI manual):
All the single couple RV cycles conform to the following pattern:
1) 33% TFA in DCM for 80 seconds
2) 50% TFA in DCM for 18.5 minutes
3) Three DCM washes
4) 10% DIEA in DMF for 1 minute
5) 10% DIEA in DMF for 1 minute
6) Five DMF washes
7) Coupling period
8) Five DCM washes
After the coupling of N-α-Boc-DL-Bhg, the Boc group is removed with the end-NH2 cycle (1.012 g).
The peptide is liberated from the solid support, and the carboxylate of aspartic acid deprotected by treatment with anhydrous hydrogen fluoride (9.0 mL), anisole (0.5 mL), and dimethyl sulfide (0.5 mL)
(60 minutes, 0°C). After removing the hydrogen fluoride under a stream of nitrogen, the resin is washed with diethyl ether (3 × 30 mL) and extracted with 20% HOAc in water (3 × 30 mL) and glacial HOAc (2 × 30 mL). The aqueous extractions are combined, concentrated under reduced pressure, and lyophilized (360 mg). The crude peptide is dissolved in 4.0 mL of 50% TFA/H2O, filtered through a 0.4 L syringe filter, and chromatographed on a Vydac 218TP 1022 column
(2.2 × 25.0 cm, 15.0 mL/min, A: 0.1% TFA/H2O,
B: 0.1% TFA/CH3CN, Gradient; 0% B for 10 minutes, 10% to 40% B over 120 minutes). Two individual fractions are collected and combined based upon analysis by analytical HPLC. The combined fractions are
concentrated separately under reduced pressure
(10 mL), diluted with H2O (50 mL), and lyophilized (40.0 mg/ea). Separation into the two diastereomers (Isomers A and B) is effected under these conditions (tR = Isomer A 15.63 min., Isomer B 16.79 min.). The late running peak fractions (Isomer B) are repurified under the same experimental conditions with a gradient of 30% to 50% B over 120 minutes at 15 mL/min to afford purified product. Acetylation is carried out with 20 mg of Isomer B in 90% acetic acid followed by addition of acetic anhydride (5 mL) and stirring overnight. After evaporation and drying the product Ac-D-Bhg-Leu-Asp-Ile-Ile-Trp is 99% pure by HPLC.
[Vydac 218 TP 1022 column (2.2 × 25.0 cm, 15.0 mL/min. A: 0.1% TFA/CH3CN, Gradient 20% to 86% B over
22 min.)] tR = 18.66 minutes. The homogeneity and structure of the resulting peptide is confirmed by analytical HPLC. Proton Nuclear Magnetic Resonance Spectroscopy (H1-NMR) and Fast Atom Bombardment Mass Spectroscopy (FAB-MS), M+Na 972.0, M+2Na+ 995.9. In a process analogous to Example 1 using the appropriate amino acids, the corresponding compounds of Formula I are prepared as follows: EXAMPLE 2
D-Bhq-Leu-Asp-Ile-Ile-Trp: FAB-MS, M+1 907.4.
EXAMPLE 3
L-Bhg-Leu-Asp-He-Ile-Trp: FAB-MS, M+1 907.4.
EXAMPLE 4
Ac-L-Bhg-Leu-Asp-lle-Ile-Trp: FAB-MS, M+1 950.0.
EXAMPLE 5
Ac-D-Txg-Leu-Asp-He-Ile-Trp; FAB-MS, M+Na 977.0.
EXAMPLE 6
Ac-D-Bheg-Leu-Asp-Ile-Ile-Trp: FAB-MS, M+1 970.3. EXAMPLE 7
Ac-D-Bhg-Orn-Asp-He-Ile-Trp: FAB-MS, M+1 951.2.
EXAMPLE 8
Ac-D-Bhg-Glu-Asp-Ile-Ile-Trp; FAB-MS, M+Na 988.8.
EXAMPLE 9
Disodium salt of Ac-D-Bhg-Leu-Asp-Ile-Ile-Trp
A saturated solution of sodium bicarbonate in water is prepared, diluted with water (1:10), chilled to 0°C, and 10 mL of the solution is added to
approximately 50 mg of Ac-D-Bhg-Leu-Asp-Ile-Ile-Trp (Example 1) with stirring. The pH of the solution is greater than 9. After 10 minutes, the solution is passed through a C18 cartridge, washed with water (100 mL), and the absorbed peptide is eluted with methanol (50 mL), concentrated under reduced pressure, resuspended in water (50 mL), and lyophilized (three times) to give the title compound.
Ac-D-Bhg-Leu-Asp-Ile-Ile-Trp, disodium salt; FAB-MS, M+1950.4, M+Na 972.1, M+2Na 994.3.
EXAMPLE 10
Boc-Bhg
Bhg-HCl (1.70 g, 5.43 mmol) is suspended in
150 mL of p-dioxane:H2O (2:1) at room temperature. To the stirred solution is added 1.40 g (6.42 mmol) of di-tert butyldicarbonate. The pH of the solution is adjusted to >9.0 with 1N NaOH and maintained at between pH 9 and 10 with aliquot additions of 1N NaOH, until the pH is constants The solution is
concentrated under reduced pressure to approximately 75 mL, overlain with ethyl acetate (50 mL), and acidified to approximately pH 2.5 with 10V aqueous HC1. The organic layer is separated, washed
successively with 10% aqueous HCl (2 × 50 mL), brine (2 × 50 mL), H2O (3 × 50 mL), and dried with MgSO4.
The solution is filtered, concentrated under reduced pressure, and the oil is recrystallized from ethyl acetate:heptane (1.82 g). The white solid is
characterized by proton NMR, fast atom bombardment mass spectrometry (M+1=368), and elemental analysis.

Claims

1. A compound of Formula I
AA1-AA2 -AA3 -AA4 -AA5-AA6
wherein AA1 is
Figure imgf000057_0002
wherein R is hydrogen,
alkyl,
alkenyl,
alkynyl,
cycloalkyl,
cycloalkylalkyl,
aryl,
heteroaryl,
fluorenylmethyl,
, wherein R2 and R3 are each the same or
Figure imgf000057_0001
different and each is
hydrogen,
alkyl,
alkenyl,
alkynyl,
cycloalkyl,
cycloalkylalkyl,
aryl,
arylalkyl, heteroaryl, or
fluorenylmethyl,
Figure imgf000058_0001
, wherein R2 is as defined above,
-OR2, wherein R2 is as defined above, , wherein R2 and R3 are as defined
Figure imgf000058_0002
above'
Figure imgf000058_0003
, wherein R9 is F, Cl, Br, or I,
-CH2-OR2, wherein R2 is as defined above. ,
Figure imgf000058_0004
wherein R2a is hydrogen or alkyl and R3 is as defined above, ,
Figure imgf000058_0005
wherein R2a and R3 are as defined above excluding R3 is hydrogen, or
Figure imgf000058_0006
, wherein R2 is as defined above,
R1 is hydrogen or alkyl,
Z is
-O-,
Figure imgf000058_0007
,
wherein m is zero or an integer of 1 or 2,
, wherein R2 is as defined above,
Figure imgf000058_0008
-(CH2)n-, wherein n is zero or an integer of 1, 2, 3, or 4, -(CH2)n-CH=CH-(CH2)n-,
wherein n is as defined above,
Figure imgf000059_0001
,
, wherein R1 and R2 are as defined
Figure imgf000059_0002
above, or ,
Figure imgf000059_0003
wherein R2 and R3 are each the same or different and each is as defined above, X and Y are the same and substituted at the same position on the aromatic ring and each may be one, two, three, or four substituents selected from the group consisting of hydrogen,
halogen,
alkyl,
-CO2R2, wherein R2 is as defined above,
, wherein R2 and R3 are as defined
Figure imgf000059_0004
above,
, wherein R2 and R3 are as defined
Figure imgf000059_0005
above, or
nitro or
Figure imgf000059_0006
wherein R, Z, X, and Y are as defined above; AA2 is
Figure imgf000060_0001
wherein R4 is
hydrogen,
alkyl,
alkenyl,
alkynyl,
cycloalkyl,
aryl,
heteroaryl,
,
Figure imgf000060_0002
wherein R2b and R3b are each the same or different and each is
hydrogen,
alkyl,
cycloalkyl,
aryl, or
heteroaryl,
-OR2b, wherein R2b is as defined above, ,
Figure imgf000060_0003
wherein R2b and R3b are each the same or different and each is as defined above for
R2b and R3b,
Figure imgf000060_0004
, wherei.n R2b is as d"efi.ned above, wherein R2b is as defined
Figure imgf000061_0001
above, or , wherein R2b is as defined
Figure imgf000061_0002
above, and
R1 and n are as defined above,
AA2 is absent;
AA3 is
Figure imgf000061_0003
wherein R5 is
hydrogen,
alkyl,
aryl,
heteroaryl, ,
Figure imgf000061_0004
wherein R2b and R3b are each the same or
different and each is as defined above, , wherein R2b is as
Figure imgf000061_0005
defined above, or , wherein R2b is as
Figure imgf000061_0006
defined above, and
R1 and n are as defined above, or
AA3 is absent; AA4 and AA5 are each independently absent or each is independently
Figure imgf000062_0004
wherein R6 is hydrogen,
alkyl,
alkenyl,
alkynyl,
cycloalkyl,
aryl, or
heteroaryl, and
R1 and n are as defined above;
AA6 is
Figure imgf000062_0003
wherein R7 is
aryl or
heteroaryl,
R8 is , wherein R1 is as defined
Figure imgf000062_0002
above,
-OR1, wherein R1 is as defined above, , wherein R1 is as defined above, or
Figure imgf000062_0001
-C^-OR1, wherein R1 is as defined
above, and
R1 and n are as defined above;
*
stereochemistry at C in AA1, AA2, AA3, AA4, or AA5 is
D, L, or DL and
*
stereochemistry at C in AA6 is L; or a
pharmaceutically acceptable salt thereof.
Figure imgf000063_0004
2. A compound according to Claim 1, in which AA1 is
Figure imgf000063_0001
wherein R is ,
Figure imgf000063_0002
wherein R2 and R3 are each the same or different and each is
hydrogen,
alkyl,
alkenyl,
alkynyl,
cycloalkyl,
cycloalkylalkyl,
aryl,
arylalkyl,
heteroaryl, or
fluorenylmethyl, , wherein R2 and R3 are as
defined above,
Figure imgf000063_0003
Figure imgf000064_0001
, wherein R9 is F, Cl, Br, or I,
Figure imgf000064_0002
, wherein R3 i.s as defined
above, or
Figure imgf000064_0003
,
wherein R3 is as defined above excluding R3 is hydrogen,
Z is -O-,
Figure imgf000064_0004
,
wherein m is zero or an integer of 1 or 2,
, wherein R2 is as defined above,
Figure imgf000064_0005
Figure imgf000064_0006
, wherein n is zero or an integer of
1, 2, 3, or 4,
-(CH2)n-CH=CH-(CH2)n-, wherein n is as
defined above,
Figure imgf000064_0007
,
, wherein R1 is hydrogen or alkyl, or
Figure imgf000064_0008
Figure imgf000064_0009
wherein R2 and R3 are each the same or different and each is as defined above and
X and Y are the same and substituted at the same position on the aromatic ring and each substituent is selected from the group consisting of
hydrogen, halogen, or
alkyl;
AA2 is
Figure imgf000065_0001
wherein R4 is hydrogen,
alkyl,
alkenyl,
alkynyl,
cycloalkyl,
aryl,
heteroaryl,
,
Figure imgf000065_0002
wherein R2b and R3b are each the same or different and each is
hydrogen,
alkyl,
cycloalkyl,
aryl, or
heteroaryl,
-OR2b, wherein R2b is as
defined above, ,
Figure imgf000065_0003
wherein R2b and R3b are each the same or different and each is as defined above for
R2b and R3b
Figure imgf000066_0006
, wherein R2b is as defined above,
Figure imgf000066_0007
wherein R2b is as
defined above, or , wherein R2b is as
Figure imgf000066_0005
defined above, and n is as defined above or
AA2 is absent;
AA3 is
Figure imgf000066_0004
wherein R5 is aryl,
heteroaryl, ,
Figure imgf000066_0003
wherein R2b and R3b are each the same or different and each is as defined above, , wherein R2b is as defined
Figure imgf000066_0002
above, or
Figure imgf000066_0001
, wherein R2b is as defined
above, and
n is as defined above, or
AA3 is absent; AA4 and AA5 are each independently absent or each is
Figure imgf000067_0003
wherein R6 is hydrogen
alkyl,
alkenyl,
alkynyl,
cycloalkyl,
aryl, or
heteroaryl, and
n is as defined above;
AA6 is
Figure imgf000067_0002
wherein R7 is aryl or
heteroaryl, and
n is as defined above, or
wherein R7, R1, and n are as
defined above,
Figure imgf000067_0001
*
stereochemistry at CH in AA1, AA2, AA3, AA4, or AA5 is
D,L, or DL, and
*
stereochemistry at CH in AA6 is L; or a
pharmaceutically acceptable salt thereof.
3. A compound according to Claim 2, in which AA1 is
Figure imgf000068_0006
wherein R is ,
Figure imgf000068_0005
wherein R2 and R3 are each the same or different and each is
hydrogen,
alkyl,
aryl, or
fluorenylmethyl, , wherein R2 and R3 are as
Figure imgf000068_0004
defined above,
Figure imgf000068_0003
, wherein R9 is F, Cl, Br,
or I, or
Figure imgf000068_0002
, wherein R10 is hydrogen, alkyl, aryl, or
arylalkyl, excluding R10 is hydrogen,
Z is -O-,
-S-,
-NH-,
- (CH2)n, wherein n is zero or an
integer of 1, 2, 3, or 4, or
,
Figure imgf000068_0001
wherein n is zero or an integer of
1 or 2 and X and Y are each the same and substituted at the same position on the aromatic ring and each substituent is selected from the group consisting of
hydrogen,
halogen, or
alkyl;
AA2 is ,
Figure imgf000069_0001
wherein R4 is hydrogen,
alkyl,
aryl,
heteroaryl,
,
Figure imgf000069_0004
wherein R2b and R3b are each the same or
different and each is hydrogen or alkyl, ,
Figure imgf000069_0002
wherein R 2b and R3b are each the same or
different . and each is hydrogen or alkyl, wherein R2b is
Figure imgf000069_0003
as defined above, or , wherein R2b is as
Figure imgf000070_0001
defined above, and n is an integer of 1, 2, 3, or 4 or
AA2 is absent;
AA3 is ,
0 )
wherein R5 is
Figure imgf000070_0002
aryl,
heteroaryl,
Figure imgf000070_0003
, wherein R3b i.s
hydrogen or
alkyl,
Figure imgf000070_0004
, wherein R2b is
hydrogen or
alkyl, or
Figure imgf000070_0005
, wherein R2b is
hydrogen or
alkyl, and
n is an integer of 1, 2, 3, or 4;
AA4 and AA5 are each independently
Figure imgf000070_0006
..
wherein R6 is hydrogen,
alkyl,
cycloalkyl, or
aryl, and n is an integer of 1, 2, 3, or 4;
AA6 is
Figure imgf000071_0001
wherein R7 is aryl or
heteroaryl, and
n is zero or an integer of 1, 2, 3, or 4, or
wherein R7, R1, and n are as
defined above,
Figure imgf000071_0002
*
stereochemistry at CH in AA 1, AA2, AA3, AA4, or AA5 is
D, L, or DL and
*
stereochemistry at CH in AA6 is L; or a
pharmaceutically acceptable salt thereof.
4. A compound according to Claim 3 selected from the group consisting of:
L-Bhg-Leu-Asp-Ile-Ile-Trp;
D-Bhg-Leu-Asp-Ile-Ile-Trp;
Ac-L-Bhg-Leu-Asp-Ile-Ile-Trp;
Ac-D-Bhg-Leu-Asp-Ile-Ile-Trp;
Ac-D-Bhg-Orn-Asp-Ile-Ile-Trp;
Ac-D-Bhg-Lys-Asp-Ile-Ile-Trp;
Ac-D-Bhg-Asp-Asp-Ile-Ile-Trp;
Ac-D-Bhg-Glu-Asp-Ile-Ile-Trp;
Ac-D-Bhg-Phe-Asp-Ile-Ile-Trp;
Ac-D-Bhg-Arg-Asp-Ile-Ile-Trp;
Ac-D-Bhg-Asp-Ile-Ile-Trp;
Fmoc-D-Bhg-Leu-Asp-Ile-Ile-Trp;
Fmoc-D-Bhg-Orn-Asp-Ile-Ile-Trp; Fmoc-D-Bhg-Lys-Asp-Ile-Ile-Trp;
Fmoc-D-Bhg-Asp-Asp-Ile-Ile-Trp;
Fmoc-D-Bhg-Glu-Asp-Ile-Ile-Trp;
Fmoc-D-Bhg-Phe-Asp-Ile-Ile-Trp; Fmc-D-Bhg-Arg-Asp-Ile-Ile-Trp;
Fmoc-D-Bhg-Asp-Ile-Ile-Trp;
Ac-D-Bhg-Leu-Phe-Ile-Ile-Trp;
Ac-D-Bhg-Leu-Asn-Ile- Ile-Trp;
Ac-D-Bhg-Leu-Glu-Ile-Ile-Trp; Ac-D-Bhg-Leu-Gln-Ile-Ile-Trp;
Ac-D-Bhg-Leu-Tyr-Ile-Ile-Trp;
Ac-D-Bhg-Leu-1-Nal-Ile-Ile-Trp;
Ac-D-Bhg-Leu-2 -Nal-Ile-Ile-Trp;
Ac-D-Bhg-Leu-Trp-Ile-Ile-Trp; Ac-D-Bhg-Leu-Asp-Val-Ile-Trp;
Ac-D-Bhg-Leu-Asp-Ile-Val-Trp;
Ac-D-Bhg-Leu-Asp-Chx-Ile-Trp;
Ac-D-Bhg-Leu-Asp-Ile-Chx-Trp;
Ac-D-Bhg-Arg-Asp-Ile-Chx-Trp; Ac-D-Bhg-Lys-Asp-Ile-Chx-Trp;
Ac-D-Bhg-Orn-Asp-Ile-Chx-Trp;
Ac-D-Bhg-Asp-Asp-Ile-Chx-Trp;
Ac-D-Bhg-Glu-Asp-Ile-Chx-Trp;
Fmoc-D-Bhg-Leu-Phe-Ile-Ile-Trp; Fmoc-D-Bhg-Leu-Asn-Ile-Ile-Trp;
Fmoc-D-Bhg-Leu-Glu-Ile-Ile-Trpr
Fmoc-D-Bhg-Leu-Gln-lie-Ile-Trp;
Fmoc-D-Bhg-Leu-Tyr-Ile-Ile-Trp;
Fmoc-D-Bhg-Leu-Asp-Val-Ile-Trp; Fmoc-D-Bhg-Leu-Asp-Ile-Val-Trp;
Fmoc-D-Bhg-Leu-Asp-Chx-Ile-Trp;
Fmoc-D-Bhg-Arg-Asp-Chx-Ile-Trp;
Fmoc-D-Bhg-Lys-Asp-Chx-Ile-Trp;
Fmoc-p-Bhg-Orn-Asp-Chx-Ile-Trp; Fmoc-D-Bhg-Asp-Asp-Chx- Ile-Trp;
Fmoc-D-Bhg-Glu-Asp-Chx-Ile-Trp;
Fmoc-D-Bhg-Leu-Asp-Ile-Chx-Trp; Fmoc-D-Bhg-Arg-Asp-Ile-Chx-Trp;
Fmoc-D-Bhg-Lys-Asp-Ile-Chx-Trp; Fmoc-D-Bhg-Orn-Asp-Ile-Chx-Trp;
Fmoc-D-Bhg-Asp-Asp-Ile-Chx-Trp;
Fmoc-D-Bhg-Glu-Asp-Ile-Chx-Trp;
Ac-D-Bheg-Leu-Asp-Ile-Ile-Trp;
Ac-D-Bheg-Orn-Asp-Ile-Ile-Trp; Ac-D-Bheg-Lys-Asp-lie-Ile-Trp;
Ac-D-Bheg-Asp-Asp-Ile-Ile-Trp;
Ac-D-Bheg-Glu-Asp-Ile-Ile-Trp;
Ac-D-Bheg-Phe-Asp-Ile-Ile-Trp;
Ac-D-Bheg-Arg-Asp-Ile-Ile-Trp; Ac-D-Bheg-Asp-Ile-Ile-Trp;
Fmoc-D-Bheg-Leu-Asp-Ile-Ile-Trp;
Fmoc-D-Bheg-Orn-Asp-Ile-Ile-Trp;
Fmoc-D-Bheg-Lys-Asp-Ile-Ile-Trp;
Fmoc-D-Bheg-Asp-Asp-Ile-Ile-Trp; Fmoc-D-Bheg-Glu-Asp-Ile-Ile-Trp;
Fmoc-D-Bheg-Phe-Asp-Ile-Ile-Trp;
Fmoc-D-Bheg-Arg-Asp-Ile-Ile-Trp;
Fmoc-D-Bheg-Asp-Ile-Ile-Trp;
Ac-D-Bheg-Leu-Phe-Ile-Ile-Trp; Ac-D-Bheg-Leu-Asn-Ile-Ile-Trp;
Ac-D-Bheg-Leu-Glu-Ile-Ile-Trp;
Ac-D-Bheg-Leu-Gln-Ile-Ile-Trp;
Ac-D-Bheg-Leu-Tyr-Ile-Ile-Trp;
Ac-D-Bheg-Leu-1-Nal-Ile-Ile-Trp; Ac-D-Bheg-Ieu-2-Nal-Ile-Ile-Trp;
Ac-D-Bheg-Leu-Trp-Ile-Ile-Trp;
Ac-D-Bheg-Leu-Asp-Val-Ile-Trp;
Ac-D-Bheg-Leu-Asp-Ile-Val-Trp;
Ac-D-Bheg-Leu-Asp-Chx-Ile-Trp; Ac-D-Bheg-Leu-Asp-Ile-Chx-Trp;
Ac-D-Bheg-Arg-Asp-Ile-Chx-Trp;
Ac-D-Bheg-Lys-Asp-Ile-Chx-Trp;
Ac-D-Bheg-Orn-Asp-Ile-Chx-Trp;
Ac-D-Bheg-Asp-Asp-Ile-Chx-Trp; Ac-D-Bheg-Glu-Asp-Ile-Chx-Trp;
Fmoc-D-Bheg-Leu-Phe-Ile-Ile-Trp;
Fmoc-D-Bheg-Leu-Asn-Ile-Ile-Trp;
Fmoc-D-Bheg-Leu-Glu-Ile-Ile-Trp;
Fmoc-D-Bheg-Leu-Gln-Ile-Ile-Trp; Emoc-D-Bheg-Leu-Tyr-Ile-Ile-Trp;
Fmoc-D-Bheg-Leu-Asp-Val-Ile-Trp;
Fmoc-D-Bheg-Leu-Asp-Ile-Val-Trp;
Fmoc-D-Bheg-Leu-Asp-Chx-Ile-Trp;
Fmoc-D-Bheg-Arg-Asp-Chx-lle-Trp;
Fmoc-D-Bheg-Lys-Asp-Chx-Ile-Trp;
Fmoc-D-Bheg-Orn-Asp-Chx-Ile-Trp;
Fmoc-D-Bheg-Asp-Asp-Chx-Ile-Trp;
Fmoc-D-Bheg-Glu-Asp-Chx-Ile-Trp;
Fmoc-D-Bheg-Leu-Asp-Ile-Chx-Trp; Fmoc-D-Bheg-Arg-Asp-Ile-Chx-Trp;
Fmoc-D-Bheg-Lys-Asp-Ile-Chx-Trp;
Fmoc-D-Bheg-Orn-Asp-Ile-Chx-Trp;
Fmoc-D-Bheg-Asp-AspτIle-Chx-Trp;
Fmoc-D-Bheg-Glu-Asp-Ile-Chx-Trp; Ac-D-Txg-Leu-Asp-Ile-Ile-Trp;
Ac-D-Txg-Orn-Asp-Ile-Ile-Trp;
Ac-D-Txg-Lys-Asp-Ile-Ile-Trp;
Ac-D-Txg-Asp-Asp-Ile-Ile-Trp;
Ac-D-Txg-Glu-Asp-Ile-Ile-Trp; Ac-D-Txg-Phe-Asp-Ile-Ile-Trp;
Ac-D-Txg-Arg-Asp-Ile-Ile-Trp;
Ac-D-Txg-Asp-Ile-Ile-Trp;
Fmoc-D-Txg-Leu-Asp-Ile- Ile-Trp;
Fmoc-D-Txg-Orn-Asp-Ile-Ile-Trp; Fmoc-D-Txg-Lys-Asp-Ile-Ile-Trp;
Fmoc-D-Txg-Asp-Asp-Ile-Ile-Trp;
Fmoc-D-Txg-Glu-Asp-Ile-Ile-Trp;
Fmoc-D-Txg-Phe-Asp-Ile-Ile-Trp;
Fmoc-D-Txg-Arg-Asp-Ile-Ile-Trp; Fmoc-D-Txg-Asp-Ile-Ile-Trp;
Ac-D-Txg-Leu-Phe-Ile-Ile-Trp; Ac-D-Txg-Leu-Asn-Ile-Ile-Trp;
Ac-D-Txg-Leu-Glu-Ile-Ile-Trp;
Ac-D-Txg-Leu-Gln-Ile-Ile-Trp; Ac-D-Txg-Leu-Tyr-Ile-Ile-Trp;
Ac-D-Txg-Leu-1-Nal-Ile-Ile-Trp;
Ac-D-Txg-Leu-2-Nal-Ile-lle-Trp;
Ac-D-Txg-Leu-Trp-Ile-Ile-Trp;
Ac-D-Txg-Leu-Asp-Val-Ile-Trp; Ac-D-Txg-Leu-Asp-Ile-Val-Trp;
Ac-D-Txg-Leu-Asp-Chx-Ile-Trp;
Ac-D-Txg-Leu-Asp-Ile-Chx-Trp;
Ac-D-Txg-Arg-Asp-Ile-Chx-Trp;
Ac-D-Txg-Lys-Asp-Ile-Chx-Trp; Ac-D-Txg-Orn-Asp-Ile-Chx-Trp;
Ac-D-Txg-Asp-Asp-Ile-Chx-Trp;
Ac-D-Txg-Glu-Asp-Ile-Chx-Trp;
Fmoc-D-Txg-Leu-Phe-Ile-Ile-Trp;
Fmoc-D-Txg-Leu-Asn-Ile-Ile-Trp; Fmoc-D-Txg-Leu-Glu-Ile-Ile-Trp;
Fmoc-D-Txg-Leu-Gln-Ile-Ile-Trp;
Fmoc-D-Txg-Leu-Tyr-Ile-Ile-Trp;
Fmoc-D-Txg-Leu-Asp-Val-Ile-Trp;
Fmoc-D-Txg-Leu-Asp-Ile-Val-Trp; Fmoc-D-Txg-Leu-Asp-Chx-Ile-Trp;
Fmoc-D-Txg-Arg-Asp-Chx-Ile-Trp;
Fmoc-D-Txg-Lys-Asp-Chx-Ile-Trp;
Fmoc-D-Txg-Orn-Asp-Chx-Ile-Trp;
Fmoc-D-Txg-Asp-Asp-Chx-Ile-Trp; Fmoc-D-Txg-Glu-Asp-Chx-Ile-Trp;
Fmoc-D-Txg-Leu-Asp-Ile-Chx-Trp;
Fmoc-D-Txg-Arg-Asp-IIe-Chx-Trp;
Fmoc-D-Txg-Lys-Asp-Ile-Chx-Trp; Fmoc-D-Txg-Orn-Asp-Ile-Chx-Trp; Fmoc-D-Txg-Asp-Asp-Ile-Chx-Trp;
Fmoc-D-Txg-Glu-Asp-Ile-Chx-Trp;
Et-D-Bhg-Leu-Asp-Ile-Ile-Trp;
Bz-D-Bhg-Leu-Asp-Ile-Ile-Trp; Pya-D-Bhg-Leu-Asp-Ile-Ile-Trp;
Cxl-D-Bhg-Leu-Asp-Ile-Ile-Trp;
Ada-D-Bhg-Leu-Asp-Ile-Ile-Trp;
Cxl(U)-D-Bhg-Leu-Asp-Ile-Ile-Trp;
Me(U)-D-Bhg-Leu-Asp-Ile-Ile-Trp;
tBu-D-Bhg-Leu-Asp-Ile-Ile-Trp;
CF3CO-D-Bhg-Leu-Asp-Ile-Ile-Trp;
Et-D-Bheg-Leu-Asp-Ile-Ile-Trp;
Bz-D-Bheg-Leu-Asp-Ile-Ile-Trp;
Pya-D-Bheg-Leu-Asp-Ile-Ile-Trp;
Cxl-D-Bheg-Leu-Asp-Ile-Ile-Trp;
Ada-D-Bheg-Leu-Asp-Ile-Ile-Trp;
Cxl(U)-D-Bheg-Leu-Asp-Ile-Ile-Trp;
Me(U) -D-Bheg-Leu-Asp-He-Ile-Trp;
tBu-D-Bheg-Leu-Asp-Ile-Ile-Trp;
CF3CO-D-Bheg-Leu-Asp-Ile-Ile-Trp;
Ac-D-Bhg-Leu-Asp-Phe-Ile-Trp;
Ac-D-Bhg-Orn-Asp-Phe-Ile-Trp;
Ac-D-Bhg-Lys-Asp-Phe-Ile-Trp;
Ac-D-Bhg-Asp-Asp-Phe-Ile-Trp;
Ac-D-Bhg-Glu-Asp-Phe-Ile-Trp;
Ac-D-Bhg-Phe-Asp-Phe-Ile-Trp;
Ac-D-Bhg-Arg-Asp-Phe-Ile-Trp;
Ac-D-Bheg-Leu-Asp-Phe-Ile-Trp;
Ac-D-Bheg-Orn-Asp-Phe-Ile-Trp;
Ac-D-Bheg-Lys-Asp-Phe-Ile-Trp;
Ac-D-Bheg-Asp-Asp-Phe-Ile-Trp;
Ac-D-Bheg-Glu-Asp-Phe-Ile-Trp;
Ac-D-Bheg-Phe-Asp-Phe-Ile-Trp; and
Ac-D-Bheg-Arg-Asp-Phe-Ile-Trp.
5. A method of inhibiting elevated levels of
endothelin comprising administering to a host suffering therefrom a therapeutically effective amount of a compound according to Claim 1 in unit dosage form.
6. A pharmaceutical composition adapted for
administration as an antagonist of endothelin comprising a therapeutically effective amount of a compound according to Claim 1 in admixture with a pharmaceutically acceptable excipient, diluent, or carrier.
7. A method of treating hypertension comprising
administering to a host suffering therefrom a therapeutically effective amount of a compound according to Claim 1 in unit dosage form.
8. A pharmaceutical composition adapted for
administration as an antihypertensive agent comprising a therapeutically effective amount of a compound according to Claim 1 in admixture with a pharmaceutically acceptable excipient, diluent or carrier.
9. A method of treating metabolic and endocrine
disorders comprising administering to a host suffering therefrom a therapeutically effective amount of a compound according to Claim 1 in unit dosage form.
10. A pharmaceutical composition adapted for
administration as an agent for treating metabolic and endocrine disorders comprising a
therapeutically effective amount of a compound according to Claim 1 in admixture with a
pharmaceutically acceptable excipient, diluent or carrier.
11. A method of treating congestive heart failure and myocardial infarction comprising administering to a host suffering therefrom a therapeutically effective amount of a compound according to
Claim 1 in unit dosage form.
12. A pharmaceutical composition adapted for
administration as an agent for treating
congestive heart failure and myocardial
infarction comprising a therapeutically effective amount of a compound according to Claim 1 in admixture with a pharmaceutically acceptable excipient, diluent or carrier.
13. A method of treating endotoxic shock comprising administering to a host suffering therefrom a therapeutically effective amount of a compound according to Claim 1 in unit dosage form.
14. A pharmaceutical composition adapted for
administration as an agent for treating endotoxic shock comprising a therapeutically effective amount of a compound according to Claim 1 in admixture with a pharmaceutically acceptable excipient, diluent or carrier.
15. A method of treating subarachnoid hemorrhage
comprising administering to a host suffering therefrom a therapeutically effective amount of a compound according to Claim 1 in unit dosage form.
16. A pharmaceutical composition adapted for
administration as an agent for treating
subarachnoid hemorrhage comprising a
therapeutically effective amount of a compound according to Claim 1 in admixture with a
pharmaceutically acceptable excipient, diluent or carrier.
17. A method of treating arrhythmias comprising administering to a host suffering therefrom a therapeutically effective amount of a compound according to Claim 1 in unit dosage form.
18. A pharmaceutical composition adapted for
administration as an agent for treating
arrhythmias comprising a therapeutically
effective amount of a compound according to
Claim 1 in admixture with a pharmaceutically acceptable excipient, diluent or carrier.
19. A method of treating asthma comprising
administering to a host suffering therefrom a therapeutically effective amount of a compound according to Claim 1 in unit dosage form.
20. A pharmaceutical composition adapted for
administration as an agent for treating asthma comprising a therapeutically effective amount of a compound according to Claim 1 in admixture with a pharmaceutically acceptable excipient, diluent or carrier.
21. A method of treating acute and chronic renal
failure comprising administering to a host suffering therefrom a therapeutically effective amount of a compound according to Claim 1 in unit dosage form.
22. A pharmaceutical composition adapted for
administration as an agent for treating acute and
. chronic renal failure comprising a
therapeutically effective amount of a compound according to Claim 1 in admixture with a
pharmaceutically acceptable excipient, diluent or carrier.
23. A method of treating preeclampsia comprising administering to a host suffering therefrom a therapeutically effective amount of a compound according to Claim 1 in unit dosage form.
24. A pharmaceutical composition adapted for
administration as an agent for treating
preeclampsia comprising a therapeutically
effective amount of a compound according to
Claim 1 in admixture with a pharmaceutically acceptable excipient, diluent or carrier.
25. A method of treating diabetes comprising
administering to a host suffering therefrom a therapeutically effective amount of a compound according to Claim 1 in unit dosage form.
26. A pharmaceutical composition adapted for
administration as an agent for treating diabetes comprising a therapeutically effective amount of a compound according to Claim 1 in admixture with a pharmaceutically acceptable excipient, diluent or carrier.
27. A method of treating neurological disorders
comprising administering to a host suffering therefrom a therapeutically effective amount of a compound according to Claim 1 in unit dosage form.
28. A pharmaceutical composition adapted for
administration as an agent for treating
neurological disorders comprising a
therapeutically effective amount of a compound according to Claim 1 in admixture with a pharmaceutically acceptable excipient, diluent, or carrier.
29. A method of treating pulmonary hypertension
comprising administering to a host suffering therefrom a therapeutically effective amount of a compound according to Claim 1 in unit dosage form.
30. A pharmaceutical composition adapted for
administration as an agent for treating pulmonary hypertension comprising a therapeutically
effective amount of a compound according to
Claim 1 in admixture with a pharmaceutically acceptable excipient, diluent, or carrier.
31. A method of treating ischemic disease comprising administering to a host suffering therefrom a therapeutically effective amount of a compound according to Claim 1 in unit dosage form.
32. A pharmaceutical composition adapted for
administration as an agent for treating ischemic disease comprising a therapeutically effective amount of a compound according to Claim 1 in admixture with a pharmaceutically acceptable excipient, diluent, or carrier.
33. A method of protecting against gastric mucosal damage or treating ischemic bowel disease
comprising administering to a host suffering therefrom a therapeutically effective amount of a compound according to Claim 1 in unit dosage form.
34. A pharmaceutical composition adapted for
administration as an agent for protecting against gastric mucosal damage or treating ischemic bowel disease comprising a therapeutically effective amount of a compound according to Claim 1 in admixture with a pharmaceutically acceptable excipient, diluent, or carrier.
35. A method of treating atherosclerotic disorders including Raynaud's disease comprising
administering to a host suffering therefrom a therapeutically effective amount of a compound according to Claim 1 in unit dosage form.
36. A pharmaceutical composition adapted for
administration as an agent for treating
atherosclerotic disorders including Raynaud's disease comprising a therapeutically effective amount of a compound according to Claim 1 in admixture with a pharmaceutically acceptable excipient, diluent, or carrier.
37. A method of treating restenosis comprising
administering to a host suffering therefrom a therapeutically effective amount of a compound according to Claim 1 in unit dosage form.
38. A pharmaceutical composition adapted for
administration as an agent for treating
restenosis comprising a therapeutically effective amount of a compound according to Claim 1 in admixture with a pharmaceutically acceptable excipient, diluent, or carrier.
39. A method of treating angina comprising
administering to a host suffering therefrom a therapeutically effective amount of a compound according to Claim 1 in unit dosage form.
40 . A pharmaceutical composition adapted for
administration as an agent for treating angina comprising a therapeutically effective amount of a compound according to Claim 1 in admixture with a pharmaceutically acceptable excipient, diluent, or carrier.
41. A method of treating cancer comprising
administering to a host suffering therefrom a therapeutically effective amount of a compound according to Claim 1 in unit dosage form.
42. A pharmaceutical composition adapted for
administration as an agent for treating cancer comprising a therapeutically effective amount of a compound according to Claim 1 in admixture with a pharmaceutically acceptable excipient, diluent, or carrier.
43. A method of treating hemorrhagic shock comprising administering to a host suffering therefrom a therapeutically effective amount of a compound according to Claim 1 in unit dosage form.
44. A pharmaceutical composition adapted for
administration as an agent for treating
hemorrhagic shock comprising a therapeutically effective amount of a compound according to
Claim 1 in admixture with a pharmaceutically acceptable excipient, diluent, or carrier.
45. A method of preparing a compound of Formula I
AA1-AA2-AA3-AA4-AA5-AA6
wherein AA1 is
Figure imgf000084_0004
wherein R is hydrogen,
alkyl,
alkenyl,
alkynyl,
cycloalkyl,
cycloalkylalkyl,
aryl,
heteroaryl,
fluorenylmethyl,
wherein R 2 and R3 are each the same or
Figure imgf000084_0003
different and each is
hydrogen,
alkyl,
alkenyl,
alkynyl,
cycloalkyl,
cycloalkylalkyl,
aryl,
arylalkyl,
heteroaryl, or
fluorenylmethyl,
Figure imgf000084_0002
, wherein R2 is as defined above, -OR2, wherein R2 is as defined above, , wherein R2 and R3 are as defined above.
Figure imgf000084_0001
Figure imgf000085_0008
, wherein R9 is F, Cl, Br, or I,
-CH2-OR2, wherein R2 is as defined above, ,
Figure imgf000085_0007
wherein R2a is hydrogen or alkyl and R3 is as defined above, ,
Figure imgf000085_0006
wherein R2a and R3 are as defined above excluding R3 is hydrogen, or
Figure imgf000085_0005
, wherein R2 is as defined above,
R1 is hydrogen or alkyl,
Z is
-O-,
Figure imgf000085_0001
,
wherein m is zero or an integer of 1 or 2,
, wherein R2 is as defined above,
Figure imgf000085_0004
-(CH2)n-, wherein n is zero or an integer of 1, 2, 3, or 4,
-(CH2)n-CH=CH-(CH2)n-,
wherein n is as defined above,
Figure imgf000085_0002
,
, wherein R1 and R2 are as defined
Figure imgf000085_0003
above, or
Figure imgf000086_0001
wherein R2 and R3 are each the same or different and each is as defined above, X and Y are the same and substituted at the same position on the aromatic ring and each may be one, two, three, or four substituents selected from the group consisting of hydrogen,
halogen,
alkyl,
-CO2R2, wherein R2 is as defined above, , wherein R and R are as defined
Figure imgf000086_0004
above,
, wherein R2 and R3 are as defined
Figure imgf000086_0003
above, or
nitro or
Figure imgf000086_0002
wherein R, Z, X, and Y are as defined above;
AA2 is
Figure imgf000086_0005
wherein R4 is
hydrogen,
alkyl, alkenyl,
alkynyl,
cycloalkyl,
aryl,
heteroaryl,
,
Figure imgf000087_0005
wherein R2b and R3b are each the same or different and each is
hydrogen,
alkyl,
cycloalkyl,
aryl, or
heteroaryl,
-OR2b, wherein R2b is as defined above,
Figure imgf000087_0004
wherein R2b and R3b are each the same or different and each is as defined above for
R2b and R3b,
Figure imgf000087_0003
, wherein R2b is as defined above,
Figure imgf000087_0002
, wherein R2b is as defined above, or
Figure imgf000087_0001
, wherein R2b is as defined above, and
R1 and n are as defined above, or
AA2 is absent; AA3 is
Figure imgf000088_0001
wherein R5 is
hydrogen,
alkyl,
aryl,
heteroaryl ,
Figure imgf000088_0002
wherein R2b and R3b are each the same or
different and each is as defined above,
Figure imgf000088_0003
R wherein R2b is as
defined above, or , wherein R2b is as
Figure imgf000088_0004
defined above, and
R1 and n are as defined above, or
AA3 is absent;
AA4 and AA5 are each independently absent or each is independently
Figure imgf000088_0005
wherein R6 is hydrogen,
alkyl,
alkenyl, alkynyl,
cycloalkyl,
aryl, or
heteroaryl, and
R1 and n are as defined above;
AA6 is
Figure imgf000089_0001
wherein R7 is
aryl or
heteroaryl,
R8 is , wherein R1 is as defined
Figure imgf000089_0002
above,
-OR1, wherein R1 is as defined above, , wherein R1 is as defined above, or
Figure imgf000089_0003
-CH2-OR1, wherein R1 is as defined above, and
R1 and n are as defined above;
*
stereochemistry at C in AA1, AA2, AA3, AA4, or AA5 is D, L, or DL and
*
stereochemistry at C in AA6 is L; or a
pharmaceutically acceptable salt thereof comprising sequential stepwise coupling of the amino acids selected from AA1, AA2, AA3, AA4, AA5, or AA6 to the preceding amino acid using conventional peptide synthesis methodology and after conventional deprotection to afford a compound of Formula I and, if desired, converting a compound of Formula I to a pharmaceutically acceptable salt of a compound of
Formula I by conventional methodology and, if further desired, converting the obtained pharmaceutically acceptable salt of a compound of Formula I to a compound of Formula I by conventional methodology.
PCT/US1993/003658 1992-04-22 1993-04-16 Endothelin antagonists ii WO1993021219A1 (en)

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EP93912310A EP0647236A1 (en) 1992-04-22 1993-04-16 Endothelin antagonists ii
AU42904/93A AU678357B2 (en) 1992-04-22 1993-04-16 Endothelin antagonists II
JP5518657A JPH07505890A (en) 1992-04-22 1993-04-16 endothelin antagonist
SK1287-94A SK128794A3 (en) 1992-04-22 1993-04-16 Peptides, method of their production and pharmaceutical agents on their base
KR1019940703745A KR950701344A (en) 1992-04-22 1993-04-16 Endothelin Antagonists II
CA002133090A CA2133090A1 (en) 1992-04-22 1993-04-16 Endothelin antagonists ii
RU9494046056A RU2100369C1 (en) 1992-04-22 1993-04-16 Peptide derivatives or their pharmaceutically acceptable salts
FI944905A FI944905A0 (en) 1992-04-22 1994-10-19 Endothelin antagonists II
NO944013A NO944013L (en) 1992-04-22 1994-10-21 Endothelin Antagonists II

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EP0626174A2 (en) 1993-04-21 1994-11-30 Takeda Chemical Industries, Ltd. Methods and compositions for the prophylactic and/or therapeutic treatment of organ hypofunction
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US6030975A (en) * 1997-03-14 2000-02-29 Basf Aktiengesellschaft Carboxylic acid derivatives, their preparation and use in treating cancer
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Cited By (20)

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US5641752A (en) * 1991-05-16 1997-06-24 Warner-Lambert Company Methods of using endothelin antagonists
US5773414A (en) * 1991-05-16 1998-06-30 Warner-Lambert Company Endothelin antagonists
US5382569A (en) * 1991-05-16 1995-01-17 Warner-Lambert Company Endotherlin antagonists
US5550110A (en) * 1992-04-22 1996-08-27 Warner-Lambert Company Endothelin Antagonists II
WO1994014843A1 (en) * 1992-12-21 1994-07-07 Warner-Lambert Company Endothelin antagonists
US6147051A (en) * 1993-04-21 2000-11-14 Takeda Chemical Industries Ltd. Methods and compositions for the prophylactic and/or therapeutic treatment of organ hypofunction
EP0626174A3 (en) * 1993-04-21 1996-01-03 Takeda Chemical Industries Ltd Methods and compositions for the prophylactic and/or therapeutic treatment of organ hypofunction.
EP0626174A2 (en) 1993-04-21 1994-11-30 Takeda Chemical Industries, Ltd. Methods and compositions for the prophylactic and/or therapeutic treatment of organ hypofunction
WO1996000738A1 (en) * 1994-06-30 1996-01-11 Warner-Lambert Company Endothelin antagonists ii
US5573762A (en) * 1995-04-24 1996-11-12 Genentech, Inc. Use of leukemia inhibitory factor specific antibodies and endothelin antagonists for treatment of cardiac hypertrophy
US6653287B1 (en) 1995-04-24 2003-11-25 Genentech, Inc. Use of leukemia inhibitory factor and endothelin antagonists
US6156733A (en) * 1995-04-24 2000-12-05 Genentech, Inc. Use of leukemia inhibitory factor and endothelin antagonists
US5837241A (en) * 1995-04-24 1998-11-17 Genentech, Inc. Method of treating heart failure using leukemia inhibitory factor antagonists optionally with endothelin antagonists
US5866568A (en) * 1995-06-07 1999-02-02 Zeneca Limited Heterocyclic compounds
WO1997033608A3 (en) * 1996-03-13 1997-11-06 Univ Kingston Antagonism of endothelin actions
US6410007B1 (en) 1996-03-13 2002-06-25 Queen's University At Kingston Antagonism of endothelin actions
US6586391B1 (en) 1996-03-13 2003-07-01 Queen's University At Kingston Method of ameliorating erectile dysfunction
WO1997033608A2 (en) * 1996-03-13 1997-09-18 Queen's University At Kingston Antagonism of endothelin actions
US6071880A (en) * 1996-09-16 2000-06-06 Dalhousie University Use of IGF-I for the treatment of renal insufficiencies, steriod toxicity and related indications
US6030975A (en) * 1997-03-14 2000-02-29 Basf Aktiengesellschaft Carboxylic acid derivatives, their preparation and use in treating cancer

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EP0647236A1 (en) 1995-04-12
NZ252855A (en) 1996-11-26
AU678357B2 (en) 1997-05-29
FI944905A0 (en) 1994-10-19
NO944013L (en) 1994-10-21
JPH07505890A (en) 1995-06-29
SK128794A3 (en) 1995-03-08
CZ256994A3 (en) 1995-02-15
AU4290493A (en) 1993-11-18
HU9403017D0 (en) 1994-12-28
KR950701344A (en) 1995-03-23

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