WO1995013817A1 - Endothelin converting enzyme inhibitors - Google Patents

Endothelin converting enzyme inhibitors Download PDF

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Publication number
WO1995013817A1
WO1995013817A1 PCT/US1994/013374 US9413374W WO9513817A1 WO 1995013817 A1 WO1995013817 A1 WO 1995013817A1 US 9413374 W US9413374 W US 9413374W WO 9513817 A1 WO9513817 A1 WO 9513817A1
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alkyl
leucyl
hydrogen
propyl
halogen
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PCT/US1994/013374
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French (fr)
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John Duncan Elliott
Chao-Pin Lee
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Smithkline Beecham Corporation
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06191—Dipeptides containing heteroatoms different from O, S, or N
    • 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 phosphorus derivatives, pharmaceutical compositions containing these compounds and their use as endothelin converting enzyme inhibitors.
  • Endothelin is a highly potent vasoconstrictor pepti.de synthesized and released by the vascular endothelium. Endothelin exists as three isoforms, ET-1, ET-2 and ET-3. [Unless otherwise stated "endothelin” shall mean any or all of the isoforms of endothelin] .
  • endothelin-1 is coded in a single gene copy as a 203 amino acid precursor, preproendothelin-1.
  • Endothelial cells contain very few secretory granules and the peptide is thought to be synthesized de novo, its production being regulated at a level of mRNA transcription.
  • Paired dibasic amino acids, at residues Lys 51 -Arg 52 anc; Arg 92 -Arg 93 ) are recognized by dibasic-pair- specific e ⁇ dopeptidases that excise a 39 amino acid residue precursor peptide, proendothelin- 1 or "big endothelin”.
  • Endothelin has profound effects on the cardiovascular system, and in particular, the coronary, renal and cerebral circulation. Elevated or abnormal release of endothelin is associated with smooth muscle contraction which is involved in the pathogenesis of cardiovascular, cerebrovascular, respiratory and renal pathophysiology. Elevated levels of endothelin have been reported in plasma from patients with essential hypertension, acute myocardial infarction,
  • endothelin has pronounced effects on blood pressure and cardiac output.
  • An intravenous bolus injection of ET (0.1 to 3 nmol/kg) in rats causes a transient, dose-related depressor response (lasting 0.5 to 2 minutes) followed by a sustained, dose-dependent rise in arterial blood pressure which can remain elevated for 2 to 3 hours following dosing.
  • Doses above 3 nmol/kg in a rat often prove fatal.
  • Endothelin appears to produce a preferential effect in the renal vascular bed. It produces a marked, long-lasting decrease in renal blood flow, accompanied by a significant decrease in GFR, urine volume, urinary sodium and potassium excretion.
  • Endothelin produces a sustained antinatriuretic effect, despite significant elevations in atrial natriuretic peptide. Endothelin also stimulates plasma renin activity.
  • ET may be an important mediator of cerebral vasospasm, a frequent and often fatal consequence of subarachnoid hemorrhage.
  • ET also exhibits direct central nervous system effects such as severe apnea and ischemic lesions which suggests that ET may contribute to the development of cerebral infarcts and neuronal death.
  • ET has also been implicated in myocardial ischemia (Nichols et al. Br. J. Pharm. 99: 597-601, 1989 and Clozel and Clozel, Circ. Res., 65: 1193-1200, 1989) coronary vasospasm (Fukuda et al. Eur. J. Pharm. 165: 301-304, 1989 and L ⁇ scher, Circ. 83: 701, 1991) heart failure, proliferation of vascular smooth muscle cells, (Takagi, Biochem & Biophvs. Res. Commun.: 168: 537-543, 1990, Bobek ej al, Am. J. Physiol. 258:408-C415, 1990) and atherosclerosis, (Nakaki et al..
  • endothelin has been found to be a potent constrictor of isolated mammalian airway tissue including human bronchus (Uchida et al.. Eur J. of Pharm. 154: 227-228 1988, LaGente, Clin. Exp. Allergy 20: 343-348, 1990; and Springall et al. Lancet. 337: 697-701, 1991).
  • Endothelin may play a role in the pathogenesis of interstitial pulmonary fibrosis and associated pulmonary hypertension, Glard et al. Third International Conference on Endothelin, 1993, p. 34 and ARDS (Adult Respiratory Distress Syndrome), Sanai et al, Supra, p. 112.
  • Endothelin has been associated with the induction of hemorrhagic and necrotic damage in the gastric mucosa (Whittle et al. Br. J. Pharm. 95: 1011-1013, 1988); Raynaud's phenomenon, Cinniniello et al. Lancet 337: 114-115, 1991); Crohn's Disease and ulcerative colitis, Munch et al.. Lancet, Vol. 339, p. 381; Migraine (Edmeads, Headache, Feb. 1991 p 127); Sepsis (Weitzberg et al., Circ. Shock 33: 222-227. 1991: Pittet et al.. Ann. Surg. 213: 262-264.
  • Endothelin stimulates both bone resorption and anabolism and may have a role in the coupling of bone remodeling. Tatrai et al Endocrinology. Vol 131, p. 603-607.
  • Endothelin has been reported to stimulate the transport of sperm in the uterine cavity, Casey et al, J. Clin. Endo and Metabolism. Vol. 74, No. 1, p. 223-225, therefore endothelin antagonists may be useful as male contraceptives.
  • Endothelin modulates the ovarian/menstrual cycle, Kenegsberg, J. of Clin. Endo. and Met.. Vol. 74, No. 1, p. 12, and may also play a role in the regulation of penile vascular tone in man, Lau et al. Asia Pacific J. of Pharm.. 1991, 6:287-292 and Tejada et al. J. Amer. Physio. Soc. 1991, H1078-H1085.
  • endothelin converting enzyme inhibitors would offer a unique approach toward the pharmacotherapy of hypertension, renal failure,
  • cerebrovascular disease myocardial ischemia, angina, heart failure, asthma, atherosclerosis, Raynaud's phenomenon, ulcers, sepsis, migraine, glaucoma, endotoxin shock, endotoxin induced multiple organ failure or disseminated intravascular coagulation, cyclosporin-induced renal failure and as an adjunct in angioplasty for prevention of restenosis, diabetes, preclampsia of pregnancy, bone remodeling, kidney transplant, male contraceptives, infertility and priaprism.
  • This invention comprises phosphorus derivatives represented by Formula (I) and pharmaceutical compositions containing these compounds, and their use as endothelin converting enzyme inhibitors which are useful in the treatment of a variety of cardiovascular and renal diseases including but not limited to: hypertension, acute and chronic renal failure, cyclosporine induced
  • nephrotoxicity nephrotoxicity, stroke, cerebrovascular vasospasm, myocardial ischemia, angina, heart failure, atherosclerosis, and as an adjunct in angioplasty for prevention of restenosis.
  • This invention further constitutes a method for inhibiting endothelin converting enzyme in an animal, including humans, which comprises administering to an animal in need thereof an effective amount of a compound of Formula (I).
  • R 1 is -(CH 2 ) n -W where n is an integer from 1-10 and W is -CONR 5 R 6 ,
  • R 5 is hydrogen orC 1-6 alkyl and R 6 is hydrogen, C 1-6 alkyl or Ar optionally substituted by OH, C 1-6 alkyl, C 1-6 alkoxy, halogen, -NHCO(C 1-6 )alkyl, -NHCOPh; or the R 5 and R 6 alkyl groups in NR 5 R 6 are joined together to form piperidine, pyrrolidine or morpholine; or W is -S(O) p -R 7 where p is 0, 1 or 2 and R 7 is C 1-6 alkyl or W is SAr;
  • q is an integer from 1 to 3, optionally substituted by one or more OH, N(R 16 ) 2 , CO 2 R 17 , halogen, or XC 1-5 alkyl groups;
  • R 2 is C 3-6 -alkyl, or CH 2 Ar;
  • R 3 is hydrogen, C 1-6 alkyl, -CH 2 -Ar wherein Ar is optionally substituted by one or more OH, N(R 16 ) 2 , CO 2 R 17 , halogen or X C 1-5 alkyl groups; or - (CH 2 ) r R 13 where r is 1 to 6, and R 13 is a piperidyl ring optionally substituted by OH, C 1-6 alkyl, C 1 -6 alkoxy, halogen,
  • R 3 is a group
  • R 14 is hydrogen, C 1-6 alkyl or -CH 2 -Ph where Ph is optionally substituted by
  • R 15 is hydrogen or C ⁇ 6alkyl
  • R 4 is hydrogen, C 1-10 alkyl, C 2-8 alkenyl, C 2-8 alkynyl, or (CH 2 ) p Ar, all of which may be unsubstituted or substituted by one or more OH, N(R 16 ) 2 ⁇ CO 2 R 17 , halogen or X C 1-5 alkyl groups;
  • R 16 is hydrogen or C 1 -4 alkyl
  • R 17 is hydrogen, C 1-6 alkyl, C 2-6 alkenyl or C 2-7 alkynyl;
  • X is (CH 2 ) n , O, N(R 16 ) or S(O) p ;
  • Ax is:
  • B is -CH 2 - or -O-;
  • R 18 is hydrogen, R 20 , OH, C 1-8 alkoxy, S(O) p R 20 , N(R 5 ) 2 , Br, F, I, Cl, CF 3 ,
  • NHCOR 5 R 20 CO 2 R 21 , XR 22 Y or X(CH 2 ) m R 23 wherein each methylene group within -X(CH 2 ) m R 23 may be unsubstituted or substituted by one or more
  • R 19 is hydrogen, R 20 , OH, C 1-5 alkoxy, S(O) p R 20 , N(R 5 ) 2 , X(R 20 ), Br , F, I, Cl or NHCOR 5 wherein the C 1 -5 alkoxy may be unsubstituted or substituted by OH, methoxy or halogen;
  • R 20 is Ar, C 1-8 alkyl, C 2-8 alkenyl, C 2-8 alkynyl all of which may be unsubstituted or substituted by one or more OH, CH 2 OH, N(R 5 ) 2 or halogen;
  • R 21 is hydrogen, C 1 - 10 alkyl, C 2- 10 alkenyl or C 2-8 alkynyl all of which may be unsubstituted or substituted by one or more OH, N(R 5 ) 2 , CO 2 R 5 , halogen or
  • XC 1-5 aIkyl or R 21 is (CH 2 ) m Ar;
  • R 23 is hydrogen, R 20 , CO 2 R 21 , CO 2 C(R 20 ) 2 O(CO)XR 21 , PO 3 (R 2 i)2.
  • R 24 is R 21 or C(R 16 ) 2 OC(O)R 4 ;
  • n is an integer from 0 to six;
  • alkyl, alkenyl, alkynyl and alkoxy groups may be straight or branched.
  • halogen is used to mean iodo, fluoro, chloro or bromo.
  • Alkyl groups may be substituted by one or more halogens up to perhalogenation.
  • the compounds of the present invention may contain one or more asymmetric carbon atoms and may exist in racemic and optically active form. All of these compounds and diastereoisomers are contemplated to be within the scope of the present invention.
  • the present invention provides compounds of Formula (I) above
  • Separation of the diastereoisomers can be achieved by high performance liquid chromatography.
  • the individual diastereoisomer can be treated with a catalyst such as palladium on activated charcoal under an atmosphere of hydrogen gas from ambient pressure to approximately 50 p.s.i., in a suitable solvent such as methanol to afford compounds of formula 1 wherein R 24 is hydrogen.
  • Compounds of Formula (I) and their pharmaceutically acceptable salts may be administered in a standard manner for the treatment of the indicated diseases, for example orally, parenterally, sub-lingually, transdermally, rectally, via inhalation or via buccal administration.
  • a syrup formulation will generally consist of a suspension or solution of the compound or salt in a liquid carrier for example, ethanol, peanut oil, olive oil, glycerine or water with a flavouring or colouring agent.
  • a liquid carrier for example, ethanol, peanut oil, olive oil, glycerine or water with a flavouring or colouring agent.
  • any pharmaceutical carrier routinely used for preparing solid formulations may be used. Examples of such carriers include magnesium stearate, terra alba, talc, gelatin, agar, pectin, acacia, stearic acid, starch, lactose and sucrose.
  • composition is in the form of a capsule
  • any routine encapsulation is suitable, for example using the aforementioned carriers in a hard gelatin capsule shell.
  • composition is in the form of a soft gelatin shell capsule
  • any pharmaceutical carrier routinely used for preparing dispersions or suspensions may be considered, for example aqueous gums, celluloses, silicates or oils and are incorporated in a soft gelatin capsule shell.
  • Typical parenteral compositions consist of a solution or suspension of the compound or salt in a sterile aqueous or non-aqueous carrier optionally containing a parenterally acceptable oil, for example polyethylene glycol, polyvinylpyrrolidone, lecithin, arachis oil, or sesame oil
  • Typical compositions for inhalation are in the form of a solution, suspension or emulsion that may be administered as a dry powder or in the form of an aerosol using a conventional propellant such as dichlorodifluoromethane or trichlorofluoromethane.
  • a typical suppository formulation comprises a compound of
  • Formula (1) or a pharmaceutically acceptable salt thereof which is active when administered in this way, with a binding and/or lubricating agent for example polymeric glycols, gelatins, cocoa-butter or other low melting vegetable waxes or fats or their synthetic analogues.
  • Typical transdermal formulations comprise a conventional aqueous or non-aqueous vehicle, for example a cream, ointment, lotion or paste or are in the form of a medicated plaster, patch or membrane.
  • the composition is in unit dosage form, for example a tablet, capsule or metered aerosol dose, so that the patient may administer to themselves a single dose.
  • Each dosage unit for oral administration contains suitably from 0.1 mg to 500 mg, and preferably from 1 mg to 100 mg, and each dosage unit for parenteral administration contains suitably from 0.1 mg to 100 mg, of a compound of Formula (I) or a pharmaceutically acceptable salt thereof calculated as the free acid.
  • Each dosage unit for intranasal administration contains suitably 1-400 mg and preferably 10 to 200 mg per person.
  • a topical formulation contains suitably 0.01 to 1.0% of a compound of Formula (I).
  • the daily dosage regimen for oral administration is suitably about 0.01 mg/Kg to 40 mg/Kg, of a compound of Formula (I) or a pharmaceutically acceptable salt thereof calculated as the free acid.
  • the daily dosage regimen for parenteral administration is suitably about 0.001 mg/Kg to 40 mg/Kg, of a compound of the Formula (I) or a pharmaceutically acceptable salt thereof calculated as the free acid.
  • the daily dosage regimen for intranasal administration and oral inhalation is suitably about 10 to about 500 mg/person.
  • the active ingredient may be administered from 1 to 6 times a day, sufficient to exhibit the desired activity.
  • Aortas were removed from freshly slaughtered pigs and immediately placed in ice-cold PBS. After rinsing three times with ice cold PBS, endothelial cells were gently scraped from the aortas with one pass of a glass slide into homogenization buffer (25 mM HEPES pH 7.5, 250 mM sucrose, ImM PMSF, 10 ⁇ M Pepstatin A, 1 mM PCMS). Cells were spun at 1000 xg for 10 minutes, resuspended in homogenization buffer and homogenized with a Polytron PT3000 for 20 sec. at 18,000 rpm.
  • homogenization buffer 25 mM HEPES pH 7.5, 250 mM sucrose, ImM PMSF, 10 ⁇ M Pepstatin A, 1 mM PCMS.
  • Peak activity fractions from the Q-Sepharose column were pooled and applied to a RCA-I lectin column equilibrated in 50 mM Tris buffer, pH 7.4, containing 0.5% Lubrol, 150 mM NaCl (Buffer B) at a linear flow rate of 15 c m/hr, at 4°C. Following a wash to baseline with Buffer B, ECE was eluted from the column with a linear gradient (0-200 mM galactose in 20 column volumes) in the same buffer. ECE activity eluted at around 20 mM galactose and the enzyme was routinely stored at -70°C.
  • Radioimmunoassay (RIA) for measuring ECE activity is a Radioimmunoassay (RIA) for measuring ECE activity
  • reaction mixture was warmed up to ambient temperature gradually and stirred at ambient temperature for 3d.
  • the reaction mixture was poured into water and extracted with CH 2 CI 2 , washed with 5% NaHCO 3 and brine, and CH 2 CI 2 solution was dried over Na 2 SO 4 and concentrated in vacuo to give a light yellow oil as a crude mixture which was subjected to reverse phase HPLC (20% H 2 O in MeOH) purification to yield 159 mg (14%) of the less polar N-[N-[1-di(phenylmethoxy)phosphinyl-3-(1,3-dioxo-1H-benz[de]isoquinoline-2-yl)propyl]-L-leucyl]-L-tryptophan phenylmethyl ester and 493 mg ( 44%) of the more polar N-[N-[l-di(phenylmethoxy)phosphinyl-3-(1,3-dioxo-1H-
  • reaction mixture was warmed up to ambient temperature gradually and stirred at ambient temperature for 3d.
  • the reaction mixture was poured into water and extracted with CH 2 CI 2 . After it was washed with 5% NaHCO 3 and brine, the CH 2 CI 2 solution was dried over Na 2 SO 4 and concentrated in vacuo to give a light yellow oil as a crude mixture which was purified by silica gel column chromatography (33% ethyl acetate in hexanes) to yield 670 mg ( 52%) of the title compound (3: 1 mixture of two diastereomers) as an oil.
  • reaction mixture was warmed up to ambient temperature gradually and stirred at ambient temperature for 3d.
  • reaction mixture was poured into water and extracted with CH 2 CI 2 . After it was washed with 5% NaHCO 3 and brine, the CH 2 CI 2 solution was dried over Na 2 SO 4 and concentrated in vacuo to give a light yellow oil as a crude mixture which was purified by silica gel column
  • reaction mixture was gradually warmed up to ambient temperature and stirred at ambient temperature for 60h.
  • the reaction mixture was poured into water and extracted with CH 2 CI 2 . After it was dried over MgSO 4 , the CH 2 CI 2 was concentrated in vacuo to give 339 mg oil as a crude mixture which was purified by silica gel column chromatography (33% hexanes in ethyl acetate) to yield 146 mg ( 36%) of the title compound ( 3: 1 mixture of two diastereomers) as an oil.
  • reaction mixture was gradually warmed up to ambient temperature and stirred at ambient temperature for 60h.
  • the reaction mixture was poured into water and extracted with CH 2 CI 2 . After it was dried over MgSO 4 , the CH 2 CI 2 was concentrated in vacuo to give 339 mg oil as a crude mixture which was purified by silica gel column chromatography ( ethyl acetate) to yield 145 mg ( 43%) of the title compound (2:1 mixture of two diastereomers) as an oil.
  • reaction mixture was warmed up to ambient temperature gradually and stirred at ambient temperature for 3d.
  • the reaction mixture was poured into water and extracted with CH 2 CI 2 . After it was washed with 0.1N HCl, 5% NaHCO 3 and brine, the CH 2 CI 2 solution was dried over Na 2 SO 4 and concentrated in vacuo giving light yellow oil as a crude mixture which was purified by silica gel column chromatography (50% ethyl acetate in hexanes) to yield 115 mg ( 13%) of the title compound as an oil.
  • Formulations for pharmaceutical use incorporating compounds of the present invention can be prepared in various forms and with numerous excipients. Examples of such formulations are given below.
  • a compound of Formula I (1 mg to 100 mg) is aerosolized from a metered dose inhaler to deliver the desired amount of drug per use.
  • Step 1 Blend ingredients No. 1, No. 2, No. 3 and No. 4 in a suitable mixer/blender.
  • Step 2 Add sufficient water portion-wise to the blend from Step 1 with careful mixing after each addition. Such additions of water and mixing until the mass is of a consistency to permit its conversion to wet granules.
  • Step 3 The wet mass is converted to granules by passing it through an oscillating granulator using a No. 8 mesh (2.38 mm) screen.
  • Step 4 The wet granules are then dried in an oven at 140°F (60°C) until dry.
  • Step 5 The dry granules are lubricated with ingredient No. 5.
  • Step 6 The lubricated granules are compressed on asuitable tablet press.
  • a pharmaceutical composition for parenteral administration is prepared by dissolving an appropriate amount of a compound of formula I in polyethylene glycol with heating. This solution is then diluted with water for injections Ph Eur. (to 100 ml). The solution is then steriled by filtration through a

Abstract

Novel phosphorous derivatives are described which are endothelin converting enzyme inhibitors.

Description

ENDOTHELIN CONVERTING ENZYME INHIBITORS
FIELD OF INVENTION
The present invention relates to novel phosphorus derivatives, pharmaceutical compositions containing these compounds and their use as endothelin converting enzyme inhibitors.
Endothelin (ET) is a highly potent vasoconstrictor pepti.de synthesized and released by the vascular endothelium. Endothelin exists as three isoforms, ET-1, ET-2 and ET-3. [Unless otherwise stated "endothelin" shall mean any or all of the isoforms of endothelin] .
In the pig, endothelin-1 is coded in a single gene copy as a 203 amino acid precursor, preproendothelin-1. Endothelial cells contain very few secretory granules and the peptide is thought to be synthesized de novo, its production being regulated at a level of mRNA transcription. Paired dibasic amino acids, at residues Lys51-Arg52 anc; Arg92-Arg93) are recognized by dibasic-pair- specific eπdopeptidases that excise a 39 amino acid residue precursor peptide, proendothelin- 1 or "big endothelin". This peptide is converted to "mature" endothelin-1 by cleavage at residues Trp73-Val74 by a neutral metalloprotease, "endothelin converting enzyme" (ECE). It has been determined that the nucleotide sequence of a partial human endothelin cDNA suggests that human endothelin-1 is derived from a 212 amino acid peptide, human preproendothelin-1, the gene for which is located on chromosome 6. The immediate precursor for "mature" human endothelin-1 is a 38 amino acid peptide.
Endothelin has profound effects on the cardiovascular system, and in particular, the coronary, renal and cerebral circulation. Elevated or abnormal release of endothelin is associated with smooth muscle contraction which is involved in the pathogenesis of cardiovascular, cerebrovascular, respiratory and renal pathophysiology. Elevated levels of endothelin have been reported in plasma from patients with essential hypertension, acute myocardial infarction,
subarachnoid hemorrhage, atherosclerosis, and patients with uraemia undergoing dialysis.
In vivo, endothelin has pronounced effects on blood pressure and cardiac output. An intravenous bolus injection of ET (0.1 to 3 nmol/kg) in rats causes a transient, dose-related depressor response (lasting 0.5 to 2 minutes) followed by a sustained, dose-dependent rise in arterial blood pressure which can remain elevated for 2 to 3 hours following dosing. Doses above 3 nmol/kg in a rat often prove fatal. Endothelin appears to produce a preferential effect in the renal vascular bed. It produces a marked, long-lasting decrease in renal blood flow, accompanied by a significant decrease in GFR, urine volume, urinary sodium and potassium excretion. Endothelin produces a sustained antinatriuretic effect, despite significant elevations in atrial natriuretic peptide. Endothelin also stimulates plasma renin activity. These findings suggest that ET is involved in the regulation of renal function and is involved in a variety of renal disorders including acute renal failure, cyclosporine nephrotoxicity and chronic renal failure.
Studies have shown that in vivo, the cerebral vasculature is highly sensitive to both the vasodilator and vasoconstrictor effects of endothelin.
Therefore, ET may be an important mediator of cerebral vasospasm, a frequent and often fatal consequence of subarachnoid hemorrhage.
ET also exhibits direct central nervous system effects such as severe apnea and ischemic lesions which suggests that ET may contribute to the development of cerebral infarcts and neuronal death.
ET has also been implicated in myocardial ischemia (Nichols et al. Br. J. Pharm. 99: 597-601, 1989 and Clozel and Clozel, Circ. Res., 65: 1193-1200, 1989) coronary vasospasm (Fukuda et al. Eur. J. Pharm. 165: 301-304, 1989 and Lϋscher, Circ. 83: 701, 1991) heart failure, proliferation of vascular smooth muscle cells, (Takagi, Biochem & Biophvs. Res. Commun.: 168: 537-543, 1990, Bobek ej al, Am. J. Physiol. 258:408-C415, 1990) and atherosclerosis, (Nakaki et al..
Biochem. & Biophys. Res. Commun. 158: 880-881, 1989, and Lerman et al.. New Eng. J. of Med. 325: 997-1001, 1991). Increased levels of endothelin have been shown after coronary balloon angioplasty (Kadel et al. No. 2491 Circ. 82: 627, 1990).
Further, endothelin has been found to be a potent constrictor of isolated mammalian airway tissue including human bronchus (Uchida et al.. Eur J. of Pharm. 154: 227-228 1988, LaGente, Clin. Exp. Allergy 20: 343-348, 1990; and Springall et al. Lancet. 337: 697-701, 1991). Endothelin may play a role in the pathogenesis of interstitial pulmonary fibrosis and associated pulmonary hypertension, Glard et al. Third International Conference on Endothelin, 1993, p. 34 and ARDS (Adult Respiratory Distress Syndrome), Sanai et al, Supra, p. 112.
Endothelin has been associated with the induction of hemorrhagic and necrotic damage in the gastric mucosa (Whittle et al. Br. J. Pharm. 95: 1011-1013, 1988); Raynaud's phenomenon, Cinniniello et al. Lancet 337: 114-115, 1991); Crohn's Disease and ulcerative colitis, Munch et al.. Lancet, Vol. 339, p. 381; Migraine (Edmeads, Headache, Feb. 1991 p 127); Sepsis (Weitzberg et al., Circ. Shock 33: 222-227. 1991: Pittet et al.. Ann. Surg. 213: 262-264. 1991), Cyclosporin-induced renal failure or hypertension (Eur. J. Pharmacol. 180: 191-192, 1990. Kidney Int. 37: 1487-1491, 1990) and endotoxin shock and other endotoxin induced diseases (Biochem. Biophys. Res. Commun., 161: 1220-1227, 1989, Acta Phvsiol. Scand. 137: 317-318, 1989) and inflammatory skin diseases. (Clin Res. 41:451 and 484, 1993).
Endothelin has also been implicated in preclampsia of pregnancy. Clark et al.. Am. J. Obstet. Gynecol. March 1992, p. 962-968; Kamor et al.. N, Eng. J. of Med.. Nov 22, 1990, p. 1486-1487; Dekker et al.. Eur J. Ob. and Gyn. and Rep. Bio. 40 (1991) 215-220; Schiff et al. Am. J. Ostet. Gynecol Feb 1992, p. 624-628; diabetes mellitus, Takahashi et_al, Diabetologia (1990) 33:306-310; and acute vascular rejection following kidney transplant, Watschinger et al.
Transplantation Vol. 52, No. 4, pp. 743-746.
Endothelin stimulates both bone resorption and anabolism and may have a role in the coupling of bone remodeling. Tatrai et al Endocrinology. Vol 131, p. 603-607.
Endothelin has been reported to stimulate the transport of sperm in the uterine cavity, Casey et al, J. Clin. Endo and Metabolism. Vol. 74, No. 1, p. 223-225, therefore endothelin antagonists may be useful as male contraceptives. Endothelin modulates the ovarian/menstrual cycle, Kenegsberg, J. of Clin. Endo. and Met.. Vol. 74, No. 1, p. 12, and may also play a role in the regulation of penile vascular tone in man, Lau et al. Asia Pacific J. of Pharm.. 1991, 6:287-292 and Tejada et al. J. Amer. Physio. Soc. 1991, H1078-H1085.
Thus, endothelin converting enzyme inhibitors would offer a unique approach toward the pharmacotherapy of hypertension, renal failure,
cerebrovascular disease, myocardial ischemia, angina, heart failure, asthma, atherosclerosis, Raynaud's phenomenon, ulcers, sepsis, migraine, glaucoma, endotoxin shock, endotoxin induced multiple organ failure or disseminated intravascular coagulation, cyclosporin-induced renal failure and as an adjunct in angioplasty for prevention of restenosis, diabetes, preclampsia of pregnancy, bone remodeling, kidney transplant, male contraceptives, infertility and priaprism.
SUMMARY OF THE INVENTION
This invention comprises phosphorus derivatives represented by Formula (I) and pharmaceutical compositions containing these compounds, and their use as endothelin converting enzyme inhibitors which are useful in the treatment of a variety of cardiovascular and renal diseases including but not limited to: hypertension, acute and chronic renal failure, cyclosporine induced
nephrotoxicity, stroke, cerebrovascular vasospasm, myocardial ischemia, angina, heart failure, atherosclerosis, and as an adjunct in angioplasty for prevention of restenosis.
This invention further constitutes a method for inhibiting endothelin converting enzyme in an animal, including humans, which comprises administering to an animal in need thereof an effective amount of a compound of Formula (I).
DETAILED DESCRIPTION OF THE INVENTION
A compound of formula I
wherein:
Figure imgf000006_0001
R1 is -(CH2)n-W where n is an integer from 1-10 and W is -CONR5R6,
-NR5COR6, -NR5CO2CH2R6 or -NR5CONR5R6 where R5 is hydrogen orC1-6alkyl and R6 is hydrogen, C1-6alkyl or Ar optionally substituted by OH, C1-6 alkyl, C1-6 alkoxy, halogen, -NHCO(C1-6)alkyl, -NHCOPh; or the R5 and R6 alkyl groups in NR5R6 are joined together to form piperidine, pyrrolidine or morpholine; or W is -S(O)p-R7 where p is 0, 1 or 2 and R7 is C1-6alkyl or W is SAr;
or W is
Figure imgf000006_0002
Figure imgf000006_0003
or
Figure imgf000007_0002
where q is an integer from 1 to 3, optionally substituted by one or more OH, N(R16)2, CO2R17, halogen, or XC1-5alkyl groups;
R2 is C3-6-alkyl, or CH2Ar;
R3 is hydrogen, C1-6alkyl, -CH2-Ar wherein Ar is optionally substituted by one or more OH, N(R16)2, CO2R17, halogen or X C1-5alkyl groups; or - (CH2)r R13 where r is 1 to 6, and R13 is a piperidyl ring optionally substituted by OH, C1-6 alkyl, C1 -6 alkoxy, halogen,
-NHCO(C1-6)alkyl, -NHCOPh or -CONR5R6; or R3 is a group
Figure imgf000007_0001
where R14 is hydrogen, C1-6alkyl or -CH2-Ph where Ph is optionally substituted by
OH, C1-6alkyl, C1-6alkoxy, halogen,-NHCO(C1-6)alkyl, -NHCOPh and
-CONR5R6; and R15 is hydrogen or Cμ6alkyl;
R4 is hydrogen, C1-10alkyl, C2-8alkenyl, C2-8alkynyl, or (CH2)pAr, all of which may be unsubstituted or substituted by one or more OH, N(R16)2· CO2R17, halogen or X C1-5alkyl groups;
R16 is hydrogen or C1 -4alkyl;
R17 is hydrogen, C1-6alkyl, C2-6alkenyl or C2-7alkynyl;
X is (CH2)n, O, N(R16) or S(O)p; Ax is:
Figure imgf000008_0001
Figure imgf000008_0002
naphthyl, indolyl, pyridyl, thienyl, oxazolidinyl, oxazolyl, thiazolyl, isothiazolyl, pyrazolyl, triazolyl, tetrazolyl, imidazolyl, imidazolidinyl, thiazolidinyl, isoxazolyl, oxadiazolyl, thiadiazolyl, morpholinyl, piperidinyl, piperazinyl, pyrrolyl, or pyrimidyl;
A is C=O or [C(R6)2]m;
B is -CH2- or -O-;
R18 is hydrogen, R20, OH, C1-8alkoxy, S(O)pR20, N(R5)2, Br, F, I, Cl, CF3,
NHCOR5, R20CO2R21, XR22Y or X(CH2)mR23 wherein each methylene group within -X(CH2)mR23 may be unsubstituted or substituted by one or more
-(CH2)mAr groups;
R19 is hydrogen, R20, OH, C1-5 alkoxy, S(O)pR20, N(R5)2, X(R20), Br, F, I, Cl or NHCOR5 wherein the C1 -5 alkoxy may be unsubstituted or substituted by OH, methoxy or halogen;
R20 is Ar, C1-8 alkyl, C2-8 alkenyl, C2-8 alkynyl all of which may be unsubstituted or substituted by one or more OH, CH2OH, N(R5)2 or halogen;
R21 is hydrogen, C1 - 10alkyl, C2- 10alkenyl or C2-8alkynyl all of which may be unsubstituted or substituted by one or more OH, N(R5)2, CO2R5, halogen or
XC1-5aIkyl or R21 is (CH2)mAr;
R22 is (CH2)m, C1 - 10alkyl C2- 10alkenyl or phenyl, all of which may be unsubstituted or substituted by one or more OH, N(R5)2, COOH, halogen, >C=O or XC1-5alkyl;
R23 is hydrogen, R20, CO2R21, CO2C(R20)2O(CO)XR21, PO3(R2i)2.
SO2NR21R20, NR21SO2R20, CONR21SO2R20, SO3R21, SO2R21,
P(O)(OR21)R21, CN, C(O)N(R5)2, tetrazole or OR5;
R24 is R21 or C(R16)2OC(O)R4;
m is an integer from 0 to six;
or a pharmaceutically acceptable salt thereof.
Also included in the invention are pharmaceutically acceptable salt complexes.
All alkyl, alkenyl, alkynyl and alkoxy groups may be straight or branched. The term "halogen" is used to mean iodo, fluoro, chloro or bromo. Alkyl groups may be substituted by one or more halogens up to perhalogenation.
The compounds of the present invention may contain one or more asymmetric carbon atoms and may exist in racemic and optically active form. All of these compounds and diastereoisomers are contemplated to be within the scope of the present invention.
The following compounds are preferred:
(IRS)-N-[N-[1-Phosphono-3-(1,3-dioxo-1H-benz[de]isoquinoline-2-yl)propyl]-L-leucyl]-L-tryptophan
(IRS)-N-[N-[1-Phosphono-3-(1,3-dioxo-1H-benz[de]isoquinoline-2-yl)propyl]-L-leucyl]-O-methyl-L-tyrosine (IRS)-N-[N-[3- Amino-1-phosphono)propyl]-L-leucyl]-L-tryptophan
(IRS)-N-[N-[1-di(Methoxy)phosphinyl-3-(1,3-dioxo-1H-benz[de]isoquinoline-2-yl)propyl]-L-leucyl]-L-tryρtophan (IRS)-N-[N-[1-Phosphono-3-(1-oxo-1H-benz[de]isoquinoline-2(3H)-yl)propyl]-L-leucyl]-L-tryptophan
(IRS)-N-[N-[1-(Methoxy)phosphono-3-(1-oxo-1H-benz[de]isoquinoline-2(3H)-yl)propyl]-L-leucyl]-L-tryptophan (IRS)-N-[N-[1-Phosphono-3-N-(N-benzyloxycarbonyl-6-aminohexanamido)propyl]-L-leucyl]-L-tryptophan
The present invention provides compounds of Formula (I) above
Figure imgf000010_0002
which can be prepared by a process which comprises: (a) reacting a dipeptidyl ester of formula 2, wherein R4 is benzyl, substituted benzyl (e.g. 4-OMe), or C1- 10 alkyl,
Figure imgf000010_0001
with an aldehyde of formula 3, in a suitable solvent such as benzene to afford an imine of formula 4.
Figure imgf000010_0003
Figure imgf000010_0004
Treatment of an imine of formula 4 with dibenzylphosphite in the presence of chlorotrimethylsilane at around 0°C, in a suitable solvent such as dichloromethane containing a base such as triethylamine, provides an alpha-aminophosphonate of formula 5, wherein Y is benzyl, as a mixture of diastereoisomers.
Figure imgf000010_0005
Separation of the diastereoisomers can be achieved by high performance liquid chromatography. The individual diastereoisomer can be treated with a catalyst such as palladium on activated charcoal under an atmosphere of hydrogen gas from ambient pressure to approximately 50 p.s.i., in a suitable solvent such as methanol to afford compounds of formula 1 wherein R24 is hydrogen.
With appropriate manipulation and protection of any chemical functionalities, synthesis of the remaining compounds of the Formula (I) is accomplished by methods analogous to those above and to those described in the Experimental section.
In order to use a compound of the Formula (I) or a pharmaceutically acceptable salt thereof for the treatment of humans and other mammals it is normally formulated in accordance with standard pharmaceutical practice as a pharmaceutical composition.
Compounds of Formula (I) and their pharmaceutically acceptable salts may be administered in a standard manner for the treatment of the indicated diseases, for example orally, parenterally, sub-lingually, transdermally, rectally, via inhalation or via buccal administration.
Compounds of Formula (I) and their pharmaceutically acceptable salts which are active when given orally can be formulated as syrups, tablets, capsules and lozenges. A syrup formulation will generally consist of a suspension or solution of the compound or salt in a liquid carrier for example, ethanol, peanut oil, olive oil, glycerine or water with a flavouring or colouring agent. Where the composition is in the form of a tablet, any pharmaceutical carrier routinely used for preparing solid formulations may be used. Examples of such carriers include magnesium stearate, terra alba, talc, gelatin, agar, pectin, acacia, stearic acid, starch, lactose and sucrose. Where the composition is in the form of a capsule, any routine encapsulation is suitable, for example using the aforementioned carriers in a hard gelatin capsule shell. Where the composition is in the form of a soft gelatin shell capsule any pharmaceutical carrier routinely used for preparing dispersions or suspensions may be considered, for example aqueous gums, celluloses, silicates or oils and are incorporated in a soft gelatin capsule shell.
Typical parenteral compositions consist of a solution or suspension of the compound or salt in a sterile aqueous or non-aqueous carrier optionally containing a parenterally acceptable oil, for example polyethylene glycol, polyvinylpyrrolidone, lecithin, arachis oil, or sesame oil Typical compositions for inhalation are in the form of a solution, suspension or emulsion that may be administered as a dry powder or in the form of an aerosol using a conventional propellant such as dichlorodifluoromethane or trichlorofluoromethane.
A typical suppository formulation comprises a compound of
Formula (1) or a pharmaceutically acceptable salt thereof which is active when administered in this way, with a binding and/or lubricating agent, for example polymeric glycols, gelatins, cocoa-butter or other low melting vegetable waxes or fats or their synthetic analogues.
Typical transdermal formulations comprise a conventional aqueous or non-aqueous vehicle, for example a cream, ointment, lotion or paste or are in the form of a medicated plaster, patch or membrane.
Preferably the composition is in unit dosage form, for example a tablet, capsule or metered aerosol dose, so that the patient may administer to themselves a single dose.
Each dosage unit for oral administration contains suitably from 0.1 mg to 500 mg, and preferably from 1 mg to 100 mg, and each dosage unit for parenteral administration contains suitably from 0.1 mg to 100 mg, of a compound of Formula (I) or a pharmaceutically acceptable salt thereof calculated as the free acid. Each dosage unit for intranasal administration contains suitably 1-400 mg and preferably 10 to 200 mg per person. A topical formulation contains suitably 0.01 to 1.0% of a compound of Formula (I).
The daily dosage regimen for oral administration is suitably about 0.01 mg/Kg to 40 mg/Kg, of a compound of Formula (I) or a pharmaceutically acceptable salt thereof calculated as the free acid. The daily dosage regimen for parenteral administration is suitably about 0.001 mg/Kg to 40 mg/Kg, of a compound of the Formula (I) or a pharmaceutically acceptable salt thereof calculated as the free acid. The daily dosage regimen for intranasal administration and oral inhalation is suitably about 10 to about 500 mg/person. The active ingredient may be administered from 1 to 6 times a day, sufficient to exhibit the desired activity.
No unacceptable toxicological effects are expected when compounds of the invention are administered in accordance with the present invention.
The biological activity of the compounds of Formula (I) are demonstrated by the following tests: Purification of ECE from Porcine Aorta Endothelial Cells
a) Solubilization of aortic endothelial cell membranes
Aortas were removed from freshly slaughtered pigs and immediately placed in ice-cold PBS. After rinsing three times with ice cold PBS, endothelial cells were gently scraped from the aortas with one pass of a glass slide into homogenization buffer (25 mM HEPES pH 7.5, 250 mM sucrose, ImM PMSF, 10μM Pepstatin A, 1 mM PCMS). Cells were spun at 1000 xg for 10 minutes, resuspended in homogenization buffer and homogenized with a Polytron PT3000 for 20 sec. at 18,000 rpm. Following centrifugation at 5000 xg for 2 min to remove cell debris, the supernatant was collected and spun at 100,000 xg for 1 hr. The pellet from the 100,000 xg spin was then resuspended in 50 mM Tris buffer, pH 7.4 containing 2% Lubrol and rotated gently at 4°C for 1 hr. After a final spin at 100,000 xg for 1 hr the majority of ECE activity (>90%) was found in the supernatant. b) Q-Sepharose Fast Flow Chromatography
The 100,000 xg supernatant from the Lubrol extract was loaded onto a Q-Sepharose Fast Flow column equilibrated in Buffer A (50 mM Tris, pH 7.4, 0.5% Lubrol) at a linear flow rate of 300 crn/hr. Following a wash with 5 column volumes of Buffer A, ECE activity was eluted with a linear salt gradient (0-600 mM NaCl in 20 column volumes) in the same buffer. ECE eluted from the column between 200 and 250 mM NaCl. c) Ricinus Communis Agarose I (RCA-1) Chromatography
Peak activity fractions from the Q-Sepharose column were pooled and applied to a RCA-I lectin column equilibrated in 50 mM Tris buffer, pH 7.4, containing 0.5% Lubrol, 150 mM NaCl (Buffer B) at a linear flow rate of 15 c m/hr, at 4°C. Following a wash to baseline with Buffer B, ECE was eluted from the column with a linear gradient (0-200 mM galactose in 20 column volumes) in the same buffer. ECE activity eluted at around 20 mM galactose and the enzyme was routinely stored at -70°C.
Radioimmunoassay (RIA) for measuring ECE activity:
Samples containing ECE activity (100 ng to 10 μg total protein) and inhibitor (10 nM - 100 μM) were incubated at 37°C for 1 hr in 50 mM
BisTrisPropane buffer, pH 7.0 following the addition of 1μM big ET-1 to initiate the reaction. After stopping the reaction by heating at 100°C for 10 min, MAb 78P, and antiserum raised against the ET-1 (15-21) peptide fragment (100 μl of a 1:25,000 dilution) and [125I]ET-1 (100 μl containing 10,000 to 15,000 cpm) were then added to 100 μl of the rection mixture diluted 20 to 1000-fold in RIA buffer (20 mM boric acid, 1 mg/ml BSA, 0.1% Tween 20, 0.002% NaN3, pH 7.4).
Following incubation for 3 hours at room temperature, 250 μl of Amerlex beads were added and the samples centrifuged at 2000 xg for 20 min. Supematants were aspirated off and the pellets counted in a gamma counter. ET-1 production in the samples was calculated by comparison to known ET-1 standards. Activites for compounds of this invention range from lnm to 50 μm. Example 1
N-[N-[1-Phosphono-3-(1,3-dioxo-1H-benz[de]isoquinoline-2-yl')propyl]-L-leucyl]-L-tryptophane a) 2-(3-Hydroxy-propyl)-2,3-dihydro-benz[de]isoquinoline-1,3-dione
10.98g (55mmol) of 1,8-naphthalic anhydride in 12.50g (170mmol) 3-aminopropanol was kept at 170°C for 30 min and 120°C for another 30 min. After it cooled to ambient temperature, the reaction mixture was dissolved in CHCI3 and washed with 1N HCL, 5% NaHCO3, and brine. The CHCI3 solution was dried over Na2SO4 and concentrated in vacuo to yield 12.14g of the title compound as light yellow solid (86%). b) 1,3-Dioxo-1H-benz[de]isoquinoline-2(3H)-propanal
To a solution of 5.00 g Dess-Martin periodinane (11.8 mmol) in 30 mL CH2CI2 was added 2.616g (10 mmol) of 2-(3-hydroxy-propyl)-2,3-dihydrobenz[de]isoquinoline-1,3-dione in 26 mL CH2CI2 over a period of 2 min at ambient temperature. After the addition was completed, the reaction was stirred at ambient temperature for 2h and 100 mL ethyl ether was added. The solution was poured into a Na2S2O3 (15g) solution in 100 mL saturated NaHCO3. After being stirred at ambient temperature for 15 min, 100 mL ethyl ether was added and the organic layer was separated. The aqueous layer was reextracted with ethyl acetate. The combined organic solutions were washed with H2O, 5% NaHCO3, and brine. After drying over Na2SO4, the solution was concentrated in vacuo to yield 2.121g of the title compound (82%) as light green solid. c) N-[N-[1-di(Phenylmethoxy)phosphinyl-3-(1,3-dioxo-1H-benz[de]isoquinoline-2-yl)propyl]-L-leucyl]-L-tryptophan phenylmethyl ester
To a solution of dibenzylphosphite (356 mg, 1.4mmol) and 138 mg (1.4 mmol) of triethyl amine in 6.2 mL CH2CI2 was added 145 mg (1.3 mmol) chlorotrimethylsilane at 0°C. After 15 min at 0°C a solution of imine, prepared previously from 312 mg (1.2 mmol) of l,3-dioxo-1H-benz[de]isoquinoline-2(3H)-propanal and 502 mg (1.2 mmol) L-leucyl-L-tryptophan phenylmethyl ester,, in 9.9 mL CH2CI2 was added at 0°C. After the addition was completed, the reaction mixture was warmed up to ambient temperature gradually and stirred at ambient temperature for 3d. For work up, the reaction mixture was poured into water and extracted with CH2CI2, washed with 5% NaHCO3 and brine, and CH2CI2 solution was dried over Na2SO4 and concentrated in vacuo to give a light yellow oil as a crude mixture which was subjected to reverse phase HPLC (20% H2O in MeOH) purification to yield 159 mg (14%) of the less polar N-[N-[1-di(phenylmethoxy)phosphinyl-3-(1,3-dioxo-1H-benz[de]isoquinoline-2-yl)propyl]-L-leucyl]-L-tryptophan phenylmethyl ester and 493 mg ( 44%) of the more polar N-[N-[l-di(phenylmethoxy)phosphinyl-3-(1,3-dioxo-1H-benz[de]isoquinoline-2-yl)propyl]-L-leucyl]-L-tryptophan phenylmethyl ester as an oil d) N-[N-[1-Phosphono-3-(1,3-dioxo-1H-benz[de]isoquinoline-2-yl)propyl]-L-leucyl]-L-tryptophane
A 15 mL MeOH solution of the less polar diastereomer of N-[N-[1-di(phenylmethoxy)phosphinyl-3-(1,3-dioxo-1H-benz[de]isoquinoline-2-yl)propyl]-L-leucyl]-L-tryptophan phenylmethyl ester (81 mg, 0.09 mmol) and a catalytic amount of Pd on activated carbon were kept under 1 atm H2 at ambient temperature for 4h. The reaction mixture was filtered through celite and concentrated to yield 39 mg (69%) of the title compound as light yellow solid. 1HNMR (CD3OD) 1.02 (d, J=6.6Hz, 6H), 1.55 (m, 1H), 1.65-2.10 (m, 4H), 3.06-3.54 (m, 4H), 3.82 (dt, J=8.8, 14.3Hz, 1H), 4.28 (dd, J=7.1, 8.6Hz, 1H), 6.77 (t, J=7.7Hz, 1H), 6.85 (t, J=7.7Hz, 1H), 7.14 (s, 1H), 7.23 (d, J=7.7Hz, 1H), 7.49 (d, J-7.7Hz, 1H), 7.82 (dd, J=7.3, 8.2Hz, 2H), 8.37 (d, 8.2Hz, 2H), 8.53 (d, J=7.3Hz, 2H); ESMS m/e 635.2 (M+l). Example 2
N-[N-[1-Phosphono-3-(1,3-dioxo-1H-benz[ de]isoquinoIine-2-yl)propyl]-L-leucyl]-L-tryptophane
Following the procedure as example Id, except substitute the more polar diastereomer of N-[N-[1-di(phenylmethoxy)phosphinyl-3-(1,3-dioxo-1H-benz[de]isoquinoline-2-y-)propyl]-L-leucyl] -L-tryptophan phenylmethyl ester for the less polar diastereomer of N-[N-[l-di(phenylmethoxy)phosphinyl-3-(1,3-dioxo-1H-benz[.ie]isoquinoline-2-yl)propyl]-L-leucyl]-L-tryptophan phenylmethyl ester (129 mg, 0.14 mmol) and the reaction were run for 3h. Seventy nine mg (87%) of the title compound was afforded as light yellow solid. 1H NMR (CD3OD) 0.95 (d, J=6.6Hz 3H), 0.97 (d, J=6.6Hz, 3H), 1.77 (m, 3H), 1.98 (m, 1H), 2.40 (m, 1H), 2.99 (m, 1H), 3.18 (dd, J=9.0, 14.3Hz, 1H), 3.40 (dd, J=5.0, 14.3Hz, 1H), 3.96 (dt, J=6.6, 13.2Hz, 1H), 4.09 (dt, J=6.6, 13.2Hz, 1H), 4.43 (dd, J=7.7, 8.8Hz, 1H), 6.99 (t, J=7.7Hz, 1H), 7.06 (t, J=7.7Hz, 1H), 7.19 (s, 1H), 7.38 (d, J=7.7Hz, 1H), 7.60 (d, J=7.7Hz, 1H), 7.75 (dd, J=7.3, 8.2Hz, 2H), 8.33 (d, J=8.2Hz, 2H), 8.52 (d, J=7.3Hz, 2H); ESMS m/e 635.2 (M+l). Example 3
N-[N-[1-Phosphono-3-(1,3-dioxo-1H-benz[de]isoquinoline-2-yl)propyl]-L-leucyl]-O -methyl-L-tyrosine a) N-[N-[1-di(Phenylmethoxy)phosphinyl-3-(1,3-dioxo-1H-benz[cie]isoquinoline-2-yl)propyl]-L-leucyl]-(9-methyl-L-tyrosine phenylmethyl ester
To a solution of dibenzylphosphite (1.139 g, 4.3mmol) and 443 mg (4.4 mmol) of triethyl amine in 20 mL CH2CI2 was added 471 mg (4.3 mmol) chlorotrimethylsilane at 0°C. After 15 min at 0°C, a solution of imine, prepared previously from 1.006 g (4.0 mmol) of l,3-dioxo-1H-benz[de]isoquinoline-2(3H)-propanal and 1.584 g (4.0 mmol) L-leucyl-(C-methyl)-L-tyrosine phenylmethyl ester in 31 mL CH2CI2 was added at 0°C. After the addition was completed, the reaction mixture was gradually warmed up to ambient temperature and stirred at ambient temperature for 3d. The reaction mixture was poured into water and extracted with CH2CI2. After it was washed with 5% NaHCO3 and brine, the CH2CI2 solution was dried over Na2SO4 and concentrated in vacuo to give a light yellow oil as a crude mixture which was subjected to reverse; phase HPLC (20% H2O in MeOH) purification to yield 520 mg ( 19%) of the less polar N-[N-[1-di(phenylmethoxy)phosphinyl-3-(1,3-dioxo-1H-benz[de]isoquinoline-2-yl)propyl]-L-leucyl]-C-methyl-L-tyrosine phenylmethyl ester and 1.56-g ( 57%) of the more polar N-[N-[1-di(phenylmethoxy)phosphinyl-3-(l, 3-dioxo-1H-benz[de]isoquinoline-2-yl)propyl]-L-leucyl]-(9-methyl-L-tyrosine phenylmethyl ester as an oil. b) N-[N-[1-Phosphono-3-(l,3-dioxo-lH-benz[de]isoquinoline-2-yl)propyl]-L-leucyl]- O-methyl-L-tyrosine
A 7.5 mL MeOH solution of the less polar diastereomer of N-[N-[1-di(phenylmethoxy)phosphinyl-3-(1,3-dioxo-1H-benz[de]isoquinoline-2-yl)propyl]-L-leucyl]- O-methyl-L-tyrosine phenylmethyl ester (39 mg, 0.04 mmol) and catalytic amount of Pd on activated carbon was kept under 1 atm H2 at ambient temperature for 5h. The reaction mixture was filted through celite and concentrated to yield 7 mg (26%) of the title compound as light yellow solid. 1HNMR (CD3OD) 0.86 (d, J=6.2Hz, 3H), 0.87 (d, J=6.2Hz, 3H), 1.32 (m, 2H), 1.67 (m, 1H), 1.92 (m, 1H), 2.20 (m, 1H), 2.72 (m, 1H), 2.84 (dd, J=9.2, 13.8Hz, 1H), 3.18 (dd, J=3.8, 13.8Hz, 1H), 3.49 (m, 1H), 3.66 (s, 3H), 3.69 (m, 1H), 4.23 (t, J=6.4Hz, 1H), 4.49 (dd, J=3.8, 9.2Hz, 1H), 6.72 (d, J=8.4Hz, 2H), 7.13 (d, J=8.4Hz, 2H), 7.78 (dd, J=7.2, 8.5Hz, 2H), 8.31 (d, J=8.5Hz, 2H), 8.52 (J=7.2Hz, 2H).
Example 4 N-[N-[1-Phosphono-3-(1,3-dioxo-1H-benz[de]isoquinoline-2-yl)propyl]-L-leucyl]-O-methyl-L-tyrosine
Following the procedure as example 3b, except substituting the more polar diastereomer of N-[N-[1-di(phenylmethoxy)phosphinyl-3-(l,3-dioxo-1H-benz[de]isoquinoline-2-yl)propyl]-L-leucyl]-6'-methyl-L-tyrosine phenylmethyl ester for the less polar diastereomer of N-[N-[l-di(phenylmethoxy)phosphinyl-3-(1,3-dioxo-1H-benz[de]isoquinoline-2-yl)propyl]-L-leucyl]-(9-methyl-L-tyrosine phenylmethyl ester (77 mg, 0.09 mmol). Thirty mg (56%) of the title compound was afforded as light yellow solid. 1H NMR (CD3OD) 0.99 (d, J=6.0Hz 3H), 1.07 (d, J=6.0Hz, 3H), 1.64 (m, 1H), 1.78 (m, 2H), 1.87 (m, 1H), 2.33 (m, 1H), 2.56 (t, J=10.2Hz, 1H), 2.86 (dd, J=10.7, 13.1Hz, 1H), 3.25 (dd, J=3.1, 13.1Hz, 1H), 3.55 (m, 1H), 3.92 (m, 1H), 4.36 (m, 1H), 4.67 (dd, J=3.1, 10.7Hz, 1H), 6.86 (d, J=8.4Hz, 2H), 7.22 (d, J=8.4Hz, 2H), 7.48 (t, J=7.4Hz, 2H), 8.04 (d, J=7.4Hz, 4H); ESMS m/e 648.2 (M+Na), 626.2 (M+1), 544.2 (M-H2PO3).
Example 5 N-[N-(3-Amino-1-phosphono)propyl]-L-leucyl]-L-tryptophan a) 3-(N-Benzyloxycarbonyl)amino-1-propanal
To a solution of 1.167 g Dess-Martin periodinane (2.8 mmol) in 7.5 mL CH2CI2 was added 518 mg (2.5 mmol) of 3-(N-benzyloxycarbonyl)amino-1-propanol in 6.4 mL CH2CI2 over a period of 1 min at ambient temperature. After the addition was completed, the reaction was stirred at ambient temperature for 30 min and 50 mL ethyl ether was added. The solution was poured into a Na2S2O3 (3.75g) solution in 25 mL saturated NaHCO3. After it was stirred at ambient temperature for 15 min, 25 mL ethyl ether was added and the organic layer was separated. The aqueous layer was reextracted with ethyl acetate. The combinet organic solutions were washed with H2O, 5% NaHCO3, and brine. After it was dried over Na2SO4, the solution was concentrated in vacuo to yield 410 mg of the title compound (80%) as white solid. 1H NMR (CDCI3) 2.73 (t, J=6.2Hz, 2H), 3.48 (dd, J=6.2, 12.4Hz, 2H), 5.08 (s, 2H), 5.15 (bs, 1H, NH), 7.34 (m, 5H), 9.79 (s, 1H). b) N-[N-[1-[di(Phenylmethoxy)phosphinyl]-3-[(phenylmethoxycarbonyl)amino]propyl]-L-leucyl]-L-tryptophan phenylmethyl ester
To a solution of dibenzylphosphite (253 mg, 1.0 mmol) and 100 mg (1.0 mmol) of triethyl amine in 4.4 mL CH2CI2 was added 105 mg (1.0 mmol) chlorotrimethylsilane at 0°C. After 15 min at 0°C a solution of imine, prepared previously from 200 mg (1.0 mmol) of 3-(N-benzyloxycarbonyl)amino-1-propanal and 393 mg (1.0 mmol) of L-leucyl-L-tryptophan phenylmethyl ester, in 8.5 mL CH2CI2 was added at 0°C. After the addition was completed, the reaction mixture was warmed up to ambient temperature gradually and stirred at ambient temperature for 3d. The reaction mixture was poured into water and extracted with CH2CI2. After it was washed with 5% NaHCO3 and brine, the CH2CI2 solution was dried over Na2SO4 and concentrated in vacuo to give a light yellow oil as a crude mixture which was purified by silica gel column chromatography (33% ethyl acetate in hexanes) to yield 670 mg ( 52%) of the title compound (3: 1 mixture of two diastereomers) as an oil. 1H NMR (CDCI3) 0.82 (d, J=6.5Hz, 6H), 1.30 (m, 3H), 1.60 (m, 2H), 2.44 (m, 1H), 2.65 (m, 1H), 2.98 (m, 1H), 3.21 (dd, J=5.5, 12.1Hz, 1H), 3.35 (dd, J=5.5, 12.1Hz, 1H), 3.69 (m, 1H), 4.83-5.15 (m, 8H), 5.38 (t, J=5.5Hz, 1H), 6.75 (bs, 1H, NH), 7.03-7.50 (m, 26H), 8.21 (bs, 1H, NH). c) N-[N-(3-Amino-1-phosphono)propyl]-L-leucyl]-L-tryptophan
A 44 mL MeOH solution of N-[N-[1-[di(phenylmethoxy)phosphinyl]-3-[(phenylmethoxycarbonyl)amino]propyl]-L-leucyl]-L-tryptophan phenylmethyl ester (223 mg, 0.26 mmol) and catalytic amount of Pd on activated carbon were kept under 1 atm H2 at ambient temperature for 30 min. The reaction mixture was filted through celite and concentrated to yield 94 mg (80%) of the title compound as white solid. *H NMR (CD3OD) 0.88 (d, J=6.6Hz 3H), 0.92 (d, J=6.6Hz, 3H), 1.29 (m, 1H), 1.43 (m, 2H), 1.61 (m, 1H), 1.90 (m, 1H), 2.42 (m, 1H), 3.08 (dd, J=11, 13.2Hz, 2H), 3.41 (dd, J=4.4, 12.1Hz, 1H), 3.50 (dd, J=3.3, 12.1Hz, 1H), 3.79 (m, 1H), 7.05 (m, 2H), 7.24 (s, 1H), 7.35 (d, J=8.8Hz, 1H), 7.64 (d, J=8.8Hz, 1H); ESMS m/e 455.2(M+1), 373.2 (M+1-H2PO3).
Example 6
N-[N-[1-di(Methoxy)phosphinyl-3-(1,3-dioxo-1H-benz[de]isoquinoline-2-yl)propyl]-L-leucyl]-L-tryptophan a) N-[N-[1-di(Methoxy)phosphinyl-3-(1,3-dioxo-1H-benz[ de]isoquinoline-2-yl)propyl]-L-leucyl]-L-tryptophan phenylmethyl ester
To a solution of dimethylphosphite (176 mg, 1.6 mmol) and 162 mg (1.6 mmol) of triethylamine in 7.4 mL CH2CI2 was added 174 mg (1.6 mmol) chlorotrimethylsilane at 0°C. After 15 min at 0°C, a solution of imine, prepared previously from 362 mg (1.4 mmol) of 1,3-dioxo-1H-benz[de]isoquinoline-2(3H)-propanal and 647 mg (1.6 mmol) of L-leucyl-L-tryptophan phenylmethyl ester, in 11 mL CH2CI2 was added at 0°C. After the addition was completed, the reaction mixture was warmed up to ambient temperature gradually and stirred at ambient temperature for 3d. For work up, the reaction mixture was poured into water and extracted with CH2CI2. After it was washed with 5% NaHCO3 and brine, the CH2CI2 solution was dried over Na2SO4 and concentrated in vacuo to give a light yellow oil as a crude mixture which was purified by silica gel column
chromatography (33% ethyl acetate in hexanes) to yield 540 mg ( 45.%) of the title compound (2:1 mixture of two diastereomers) as an oil. b) N-[N-[1-di(Methoxy)phosphinyl-3-(1,3-dioxo-1H-benz[de]isoquinoline-2-yl)propyl]-L-leucyl]-L-tryptophan
A 30 mL MeOH solution of N-[N-[1-di(methoxy)phosphinyl-3-(1,3-dioxo-1H-benz[de]isoquinoline-2-yl)propyl]-L-leucyl]-L-tryptophan phenylmethyl ester (286 mg, 0.38 mmol) and catalytic amount of Pd on activated carbon were kept under 1 atm H2 at ambient temperature for 5h. The reaction mixture was filtered through celite and concentrated to yield 222 mg (88%) of the title compound as light yellow solid. 1H NMR (CD3OD) 0.88 (d, J=6.6Hz, 6H), 1.28 (m, 2H), 1.70 (m, 2H), 1.93 (m, 1H), 3.00 (m, 1H), 3.38 (m, 2H), 3.64 (d, J=11.0Hz, 3H), 3.70 d, J-11.0Hz, 3H), 3.78 (m, 1H), 3.95 (m, 1H), 4.10 (t, J-9.4Hz, 1H), 6.77 (dd, J=9.9, 9.9Hz, 1H), 6.88 (dd, J=8.8, 9.9Hz, 1H), 7.09 (d, J=8.8Hz, 1H), 7.15 (s, 1H), 7.57 (d, J=9.9Hz, 1H), 7.77 (dd, J=7.7, 13.2Hz, 2H), 8.29 (d, J=7.7Hz, 2H), 8.46 (d, J= 13.2Hz, 2H). Example 7
N-[N-[1-Phosphono-3- (1-oxo-1H-benz[de]isoquinoline-2(3H)-yl)propyl]-L-leucyl]-L-tryptophane a) 2-(3-Hydroxy-propyl)-1H-benz[de]isoquinoline-1(3H)-one
To a solution of 1.276g (5 mmol) of 2-(3-hydroxy-propyl)-2,3-dihydrobenz[de]isoquinoline-l,3-dione in 200 mL 9% H2O in EtOH was added 1.892g (50 mmol) NaBH4 at ambient temperature and stirred at ambient temperature for 4h.
After treatment with 1N HCl, the reaction mixture was extracted with CH2CI2. The CH2CI2 was dried over Na2SO4 and concentrated in vacuo to yield a crude mixture which was subjected to silica gel column chromatography (ethyl acetate) to yield
369 mg (30%) of the title compound as an oil b) 1-Oxo-1H-benz[de]isoquinoline-2(3H)-propanal
To a solution of 1.059 g Dess-Martin periodinane (2.5 mmol) in 6 mL
CH2CI2 was added 548 mg (2.3 mmol) of 2-(3-hydroxy-propyl)-1H-benz[de]isoquinoline-1(3H)-one in 5 mL CH2CI2 over a period of 1 min at ambient temperature. After the addition was completed, the reaction was stirred at ambient temperature for 1h and 13 mL ethyl ether was added. The solution was poured into a Na2S2O3 (3.28g) solution in 13 mL saturated NaHCO3. After it was stirred at ambient temperature for 15 min, 25 mL ethyl acetate was added and the organic layer was separated. The aqueous layer was reextracted with ethyl acetate. The combined organic solutions were dried over Na2SO4 and concentrated in vacuo to give a crude mixture which was purified by silica gel column chromatography (ethyl acetate) to yield 276 mg of the title compound (51%) as light yellow solid. 1H NMR (CDCl3) 2.80 (dd, J=1.1, 6.2Hz, 2H), 3.74 (t, J=6.4Hz, 2H), 4.77 (s, 2H), 7.09 (dd, J=1.1, 7.1Hz, 1H), 7.27 (dd, J=7.2, 8.1Hz, 1H), 7.36 (dd, J=7.2, 8.1Hz, 1H), 7.54 (dd, J=0.8, 8.3Hz, 2H), 7.72 (dd, J=0.8. 8.3Hz, 2H), 8.11 (d, J=7.0Hz, 1H), 9.72 (t, J=1.1Hz, 1H). c) N-[N-[3-(1-Oxo-1H-benz[de]isoquinoline-2(3H)-yl)propylidene]-L-leucyl]-L-tryptophane phenylmethyl ester
To a solution of trifluoroacetate salt of L-leucyl-L-tryptophan phenylmethyl ester (478 mg, 0.9 mmol) in 6 mL CH2CI2 was added 93 mg (0.9 mmol) of triethylamine, 219 mg of 1-oxo-1H-benz[de]isoquinoline-2(3H)-propanal (0.9 mmol) in 3.3 mL CH2CI2, and MgSO4 at 0°C. After it was warmed to ambient temperature, the solution was kept at ambient temperature for 15 min. After MgSO4 was filtered out, the solution was concentrated in vacuo to yield 576 mg (100%) of the title compound as an oil. d) N-[N-[1-di(Phenylmethoxy)phosphinyl-3-(1-Oxo-1H-benz[de]isoquinoline-2(3H)-yl)propyl]-L-leucyl]-L-tryptophane phenylmethyl ester
To a solution of dibenzylphosphite (132 mg, 0.5 mmol) and 51 mg (0.5 mmol) of triethyl amine in 1.4 mL CH2CI2 was added 55 mg (0.5 mmol) chlorotrimethylsilane at 0°C. After 15 min at 0°C, 288 mg (0.5 mmol) of N-[N-[3-(l-oxo-lH-benz[de]isoquinoline-2(3H)-yl)propylidene]-L-leucyl]-L-tryptophane phenylmethyl ester in 1 mL CH2CI2 was added at 0°C. After the addition was completed, the reaction mixture was gradually warmed up to ambient temperature and stirred at ambient temperature for 60h. The reaction mixture was poured into water and extracted with CH2CI2. After it was dried over MgSO4, the CH2CI2 was concentrated in vacuo to give 339 mg oil as a crude mixture which was purified by silica gel column chromatography (33% hexanes in ethyl acetate) to yield 146 mg ( 36%) of the title compound ( 3: 1 mixture of two diastereomers) as an oil. 1H NMR (CDCI3) of less polar diastereomer 0.78 (d, J=6.6Hz, 3H), 0.80 (d, J=6.6Hz, 3H), 1.20-1.69 (m, 3H), 1.84 (m, 1H), 2.01 (m, 1H), 2.88 (m, 1H), 3.01 (m, 1H, NH), 3.22 (m, 1H), 3.34 (dd, J=3.3, 12.1Hz, 1H), 3.55 (m, 2H), 4.68 and 4.73 (ABq, J=14.3Hz, 2H), 4.80-5.14 (m, 9H), 6.76-7.58 (m, 22H), 7.74 (d, J=8.8Hz, 1H), 7.77 (d, J=6.6Hz, 1H), 7.94 (d, J=8.8Hz, 1H), 8.30 (d, J=6.6Hz, 1H), 8.38 (bs, 1H, NH);1H NMR (CDCI3) of more polar diastereomer 0.83 (d, J=5.5Hz, 3H), 0.85 (d, J=5.5Hz, 3H), 1.30 (m, 1H), 1.45 (m, 1H), 1.60 (m, 3H), 2.88 (m, 2H), 3.19(dd, J=5.5, 14.3Hz, 1H), 3.53 (m, 2H), 3.76 (m, 1H), 4.15 (d, J=16.5Hz, 1H), 4.50 (d, J=16.5Hz, 1H), 4.80-5.14 (m, 9H), 6.78 (t, J=8.8Hz, 1H), 6.87 (d, J=8.8Hz, 1H), 6.93-7.33 (m, 18H), 7.45 (d, J=9.9Hz, 1H), 7.51 (t, J=9.9Hz, 1H), 7.63 (t, J=9.9Hz, 1H), 7.81 (d, J=9.9Hz, 1H), 8.00 (d, J=9.9Hz, 1H), 8.39 (d, J=9.9Hz, 1H), 8.98 (bs, 1H, NH). e) N-[N-[1-Phosphono-3-(1-oxo-1H-benz[de]isoquinoline-2(3H)-yl)propyl]-L-leucyl]-L-tryptophane
A 7 mL MeOH solution of N-[N-[l-di(phenylmethoxy)phosphinyl-3-(1-oxo- 1H-benz[de]isoquinoline-2(3H)-yl)propyl]-L-leucyl]-L-tryptophane phenylmethyl ester (69 mg, 0.08 mmol) and catalytic amount of Pd on activated carbon were kept under 1 atm H2 at ambient temperature for 5.5h. The reaction mixture was filted through celite and concentrated to yield 47 mg (83%) of the title compound as a white forming solid. 1H NMR (CD3OD) 0.84 (d, J=6.6Hz 3H), 0.88 (d, J=6.6Hz, 3H), 1.31 (m, 1H), 1.43 (m, 1H), 1.71 (m, 2H), 1.96 (m, 1H), 2.54 (dt, J=3.3, 11Hz, 1H), 3.22 (dd, J=6.6, 13.2Hz, 1H), 3.36-3.68 (m, 3H), 4.52 (t, J=6.6Hz, 1H), 4.82 (s, 2H), 6.87 (m, 2H), 7.14 (s, 1H), 7.19 (d, J=7.7Hz, 1H), 7.37 (d, J=7.7Hz, 1H), 7.49 (m, 2H), 7.60 (d, J=8.8Hz, 1H), 7.73 (d, J=8.8Hz, 1H), 7.92 (d, J=8.8Hz, 1H), 8.13 (d, J=8.8Hz, 1H); ESMS m/e 621.2 (M+l).
Example 8
N-[N-[1-(Methoxy)phosphono-3-(1-oxo-1H-benz[ de]isoquinoline-2(3H)-yl)propyl]- L-leucyl]-L-tryptophane a) N-[N-[1-di(Methoxy)phosphinyl-3-(1-oxo-1H-benz[ de]isoquinoline-2(3H)-yl)propyl]-L-leucyl]-L-tryptophane phenylmethyl ester
To a solution of dimethylphosphite (55 mg, 0.5 mmol) and 51 mg (0.5 mmol) of rriethyl amine in 1.4 mL CH2CI2 was added 55 mg (0.5 mmol) chlorotrimethylsilane at 0°C. After 15 min at 0°C, 288 mg (0.5 mmol) of N-[N-[3-(1-oxo-1H-benz[de]isoquinoline-2(3H)-yl)propylidene]-L-leucyl]-L-tryptophane phenylmethyl ester in 1 mL CH2CI2 was added at 0°C. After the addition was completed, the reaction mixture was gradually warmed up to ambient temperature and stirred at ambient temperature for 60h. The reaction mixture was poured into water and extracted with CH2CI2. After it was dried over MgSO4, the CH2CI2 was concentrated in vacuo to give 339 mg oil as a crude mixture which was purified by silica gel column chromatography ( ethyl acetate) to yield 145 mg ( 43%) of the title compound (2:1 mixture of two diastereomers) as an oil. 1H NMR (CDCI3) of less polar diastereomer 0.80 (d, J=6.6Hz, 3H), 0.82 (d, J=6.6Hz, 3H), 1.33 (m, 1H), 1.46 (m, 1H), 1.65 (m, 3H), 2.86 (m, 2H), 3.32 (dd, J=8.8, 11.0Hz, 1H), 3.45 (t, J=6.6Hz, 1H), 3.50-3.68 (m, 8H), 4.82 and 4.88 (ABq, J=17.6Hz, 2H), 4.90-5.09 (m, 5H), 6.78-7.74 (m, 14H), 7.91 (d, J=8.8Hz, 1H), 8.28 (d, J=8.8Hz, 1H), 8.92 (bs, 1H, NH); 1H NMR (CDCI3) of more polar diastereomer 0.92 (d, J=8.8Hz, 3H), 0.93 (d, J=8.8Hz, 3H), 1.33 (m, 1H), 1.46 (m, 1H), 1.65 (m, 3H), 2.86 (m, 2H), 3.19(dd, J=5.5, 13.2Hz, 1H), 3.53 (m, 2H), 3.60 (d, J=11Hz, 3H), 3.65 (d, J=11Hz, 3H), 4.23 (d, J=16.5Hz, 1H), 4.62 (d, J=16.5Hz, 1H), 5.01 (s, 2H), 5.04 (m, 3H), 6.79 (t, J=8.8Hz, 1H), 6.90-7.55 (m, 11H), 7.58 (t, J=7.7Hz, 1H), 7.77 (d, J=9.9Hz, 1H), 7.94 (d, J=9.9Hz, 1H), 8.35 (d, J=7.7Hz, 1H), 9.12 (bs, 1H, NH). b) N- [N-[1-di(Methoxy)phosphinyl-3-(1-oxo-1H-benz[de]isoquinoline-2(3H)-yl)propyl]-L-leucyl]-L-tryptophane
A 5.5 mL MeOH solution of N-[N-[1-di(methoxy)phosphinyl-3-(1-oxo-1H-benz[de]isoquinoline-2(3H)-yl)proρyl]-L-leucyl]-L-tryptophane phenylmethyl ester (45 mg, 0.06 mmol) and catalytic amount of Pd on activated carbon were kept under 1 atm H2 at ambient temperature for 30 min. The reaction mixture was filtered through celite and concentrated to yield 34 mg (86%) of the title compound as a white forming solid. 1H NMR (CD3OD) 0.89 (d, J=7.7Hz 3H), 0.91 (d, J=7.7Hz, 3H), 1.33 (m, 3H), 1.71 (m, 2H), 3.03 (m, 1H), 3.26-3.58 (m, 3H), 3.68 (d, J=11.0Hz, 3H), 3.72 (d, J=11.0Hz, 3H), 4.60 (d, 16.5Hz, 2H), 5.00-5.15 (m, 1H), 6.75 (t, J=8.8Hz, 1H), 6.84 (t, 8.8Hz, 1H), 7.05 (d, J=8.8Hz, 1H), 7.10 (s, 1H), 7.39 (d, J=8.8Hz, 1H), 7.50-7.63 (m, 3H), 7.83 (d, J=9.9Hz, 1H), 8.02 (d, J=9.9Hz, 1H), 8.17 (d, J=9.9Hz, 1H). c) N-[N-[1-(Methoxy)phosphono-3-(1-oxo-1H-benz[de]isoquinoline-2(3H)-yl)propyl]-L-leucyl]-L-tryptophane
To a 1 mL MeOH solution of 5 mg (0.01 mmol) of N-[N-[1-di(methoxy)phosphinyl-3-(1-oxo-1H-benz[de]isoquinoline-2(3H)-yl)propyl]-L-leucyl]-L-tryptophane was added 1N KOH at ambient temperature. After it was stirred at ambient for 2h, the solution was concentrated in vacuo to give 3.5 mg (64%) of the title compound as solid. 1H NMR (CD3CD) 0.88 (d, J=8.2Hz, 6H), 1.31 (m, 3H), 1.69 (m, 1H), 1.97 (m, 1H), 2.70 (m, 1H), 3.30-3.50 (m, 2H), 3.55 (d, J=11.0Hz, 3H), 4.58 (t, J=7.1Hz, 1H), 5.05-5.16 (m, 5H), 6.83 (m, 2H), 7.12 (s, 1H), 7.13 (dd, J=8.2, 8.2Hz, 1H), 7.41 (d, J=8.2Hz, 1H), 7.57 (m, 3H), 7.80 (d, J=9.7Hz, 1H), 7.99 (d, J=10.4Hz, 1H), 8.17 (d, J=10.4Hz, 1H). Example 9
N-rN-ri-Phsophono-3-N-(N-benzyloxycarbonyl-6-aminohexanamido)propyl]-L-leucyl]-L-tryptophane a) N-Benzyloxycarbonyl-6-amino-N-(3-hydroxy-proρyl)-hexanamide
To a solution of 482 mg (1.3 mmol) of the N-benzyloxycarbonyl-6-aminohexanoyl N-hydroxysuccimide ester in 2.9 mL DMF was added 91 mg (1.2 mmol) of 3-amino- 1-propanol at ambient temperature and stirred at ambient temperature for 14h. After the DMF was removed in vacuo, the resulting crude mixture was redissolved in ethyl acetate and washed with water, 1N HCl, and 5% NaHCO3. After it was dried over MgSO4, the ethyl acetate was concentrated in vacuo to give 241 mg (56%) of the title compound as white solid. 1H NMR (CDCI3) 1.35 (dt, J=7.2, 14.4Hz, 2H), 1.53 (dt, J=8.2, 16.4Hz, 2H), 1.68 (m, 4H), 2.20 (t, J=8.2Hz, 2H), 3.19 (dd, J=8.8, 13.2Hz, 2H), 3.28 (bs, 1H, OH), 3.42 (dd, J=7.2, 11.0Hz, 2H), 3.64 (dd, J=4.4, 11.0Hz, 2H), 4.84 (bs, 1H, NH), 5.10 (s, 2H), 5.93 (bs, 1H, NH), 7.35 (m, 5H). b) 3-Amino-N-(N-benzyloxycarbonyl-6-aminohexanoyl)propanal
To a solution of 530 mg Dess-Martin periodinane (1.2 mmol) in 3 mL CH2CI2 was added 365 mg (1.1 mmol) of N-benzyloxycarbonyl-6-amino-N-(3-hydroxy-propyl)-hexanamide in 5 mL CH2CI2 over a period of 1 min at ambient temperature. After the addition was completed, the reaction was stirred at ambient temperature for lh and 7.5 mL ethyl ether was added. The solution was poured into a Na2S2O3 (1.64g) solution in 7.5 mL saturated NaHCO3. After it was stirred at ambient temperature for 5 min, the solution was extracted with ethyl acetate. The organic solution was washed with 5% NaHCO3, and dried over MgSO4. The solution was concentrated in vacuo to yield 292 mg of the title compound (83%) as an oil. c) N- [N-[1-di(Phenylmethoxy)phosphinyl-3-N-(N-benzyloxycarbonyl-6-aminohexanamido)propyl]-L-leucyl] -L-tryptophane phenylmethyl ester
To a solution of dibenzylphosphite (263 mg, 1.0 mmol) and 100 mg (1.0 mmol) of triethyl amine in 4.5 mL CH2CI2 was added 110 mg (1.0 mmol) chlorotrimethylsilane at 0°C. After 15 min at 0°C 620 mg (0.9 mmol) of imine, prepared previously from 292 mg (0.9 mmol) of 3-amino-N*(N-benzyloxycarbonyl- 6-aminohexanoyl)propanal and 370 mg (0.9 mmol) of L-leucyl-L-tryptophan phenylmethyl ester, was added at 0°C. After the addition was completed, the reaction mixture was warmed up to ambient temperature gradually and stirred at ambient temperature for 3d. The reaction mixture was poured into water and extracted with CH2CI2. After it was washed with 0.1N HCl, 5% NaHCO3 and brine, the CH2CI2 solution was dried over Na2SO4 and concentrated in vacuo giving light yellow oil as a crude mixture which was purified by silica gel column chromatography (50% ethyl acetate in hexanes) to yield 115 mg ( 13%) of the title compound as an oil. d) N-[N-[l-Phsophono-3-N-(N-benzyloxycarbonyl-6-aminohexanamido)propyl]-L-leucyl]-L-tryptophane
A 15 mL MeOH solution of N-[N-[l-di(phenylmethoxy)phosphinyl-3-N-(N-benzyloxycarbonyl-6-aminohexanamido)propyl]-L-leucyl]-L-tryptophane phenylmethyl ester (115 mg, 0.12 mmol) and catalytic amount of Pd on activated carbon were kept under 1 atm H2 at ambient temperature for 5h. The reaction mixture was filtered through celite and concentrated to yield 60 mg (89%) of the title compound as pink solid.1H NMR (CD3OD) 0.92 (d, J=6.6Hz 3H), 0.98 (d, J=6.6Hz, 3H), 1.31 (dt, J=8.2, 16.5Hz, 2H), 1.64 (m, 9H), 2.10 (t, J=8.8Hz, 2H), 2.39 (m, 1H), 2.70 (m, 1H), 2.89 (t, J=9.9Hz, 2H), 3.00 (m, 1H), 3.15 (dd, J=11.0, 13.2Hz, 1H), 3.46 (dd, J=4.4, 13.2Hz, 1H), 4.48 (m, 1H), 4.83 (dd, J=4.4, 11.0Hz, 1H), 7.03 (t, J=8.8Hz, 1H), 7.10 (t, J=8.8Hz, 1H), 7.24 (s, 1H), 7.39 (d, J=8.8Hz, 1H), 7.61 (d, J=8.8Hz, 1H); ESMS m/e 568.4(M+1). EXAMPLE 10
Formulations for pharmaceutical use incorporating compounds of the present invention can be prepared in various forms and with numerous excipients. Examples of such formulations are given below.
Inhalant Formulation
A compound of Formula I, (1 mg to 100 mg) is aerosolized from a metered dose inhaler to deliver the desired amount of drug per use. Tablets/Ingredients Per Tablet
1. Active ingredient 40 mg
(Cpd of Form. I)
2. Corn Starch 20 mg
3. Alginic acid 20 mg
4. Sodium Alginate 20 mg
5. Mg stearate 1.3 mg
2.3 mg Procedure for tablets:
Step 1 Blend ingredients No. 1, No. 2, No. 3 and No. 4 in a suitable mixer/blender.
Step 2 Add sufficient water portion-wise to the blend from Step 1 with careful mixing after each addition. Such additions of water and mixing until the mass is of a consistency to permit its conversion to wet granules.
Step 3 The wet mass is converted to granules by passing it through an oscillating granulator using a No. 8 mesh (2.38 mm) screen.
Step 4 The wet granules are then dried in an oven at 140°F (60°C) until dry.
Step 5 The dry granules are lubricated with ingredient No. 5.
Step 6 The lubricated granules are compressed on asuitable tablet press.
Parenteral Formulation
A pharmaceutical composition for parenteral administration is prepared by dissolving an appropriate amount of a compound of formula I in polyethylene glycol with heating. This solution is then diluted with water for injections Ph Eur. (to 100 ml). The solution is then steriled by filtration through a
0.22 micron membrane filter and sealed in sterile containers.

Claims

CLAIMS:
1. A compound of formula I
Figure imgf000027_0004
wherein:
R1 is -(CH2)n-W where n is an integer from 1-10 and W is -CONR5R6,
-NR5COR6, -NR5CO2CH2R6 or -NR5CONR5R6 where R5 is hydrogen or C1-5alkyl and R6 is hydrogen, C1-6alkyl or Ar optionally substituted by OH, C1-6 alkyl, C1-6 alkoxy, halogen, -NHCO(C1-6)alkyl, -NHCOPh; or the R5 and R6 alkyl groups in NR5R6 are joined together to form piperdine, pyrrolidine or morpholine; or W is -S(O)p-R7 where p is 0, 1 or 2 and R7 is C1-6alkyl or W is SAr;
or W is
;
Figure imgf000027_0001
Figure imgf000027_0002
or
Figure imgf000027_0003
where q is an integer from 1 to 3, optionally substituted by one or more OH, N(R16)2, CO2R17, halogen, or XC1-5alkyl groups; R2 is C3-6alkyl or -CH2-Ar;
R3 is hydrogen, C1-6alkyl, -CH2-Ar wherein Ar is optionally substituted by one or more OH, N(R16)2, CO2R17, halogen or XC1-5alkyl groups; or - (CH2)r-R13 where r is 1 to 6, R13 is a piperidyl ring optionally substituted by OH, C1-6 alkyl, C1-6 alkoxy, halogen,
-NHCO(C1-6)alkyl, -NHCOPh or -CONR5R6; or R3 is a group
Figure imgf000028_0003
where R1 4 is hydrogen, C1-6alkyl or -CH2-Ph where Ph is optionally substituted by OH, C1-6alkyl, C1-6alkoxy, halogen,-NHCO(C1-6)alkyl, -NHCOPh and
-CONR5R6; and R15 is hydrogen or C1-6alkyl;
R4 is hydrogen, C1-6alkyl, C2-8alkenyl, C2-8alkynyl, or (CH2)pAr, all of which may be unsubstituted or substituted by one or more OH, N(R16)2. CO2R17, halogen orXC1-5alkyl groups;
R16 is hydrogen or C1-4alkyl;
R17 is hydrogen, C1-6alkyl, C2-6alkenyl or C2-7alkynyl;
X is (CH2), O, NR16 or S(O)p:
Ar is:
Figure imgf000028_0002
Figure imgf000028_0001
naphthyl, indolyl, pyridyl, thienyl, oxazolidinyl, oxazolyl, thiazolyl, isothiazolyl, pyrazolyl, triazolyl, tetrazolyl, imidazolyl, imidazolidinyl, thiazolidinyl, isoxazolyl, oxadiazolyl, thiadiazolyl, morpholinyl, piperidinyl, piperazinyl, pyrrolyl, or pyrimidyl;
A is C=O or [C(R6)2]m;
B is -CH2- or -O-;
R18 is hydrogen, R20, OH, C1-8alkoxy, S(O)pR20, N(R5)2, Br, F, I, Cl, CF3,
NHCOR5, R20CO2R21, XR22Y or X(CH2)mR23 wherein each methylene group within -X(CH2)mR23 may be unsubstituted or substituted by one or more
-(CH2)rr-Ar groups;
R19 is hydrogen, R20, OH, C1-5 alkoxy, S(O)pR20, N(R5)2, X(R20), Br, F, I, Cl or NHCOR5 wherein the C1-5 alkoxy may be unsubstituted or substituted by OH, methoxy or halogen;
R20 is Ar, C1-8 alkyl, C2-8 alkenyl, C2-8 alkynyl all of which may be unsubstituted or substituted by one or more OH, CH2OH, N(R5)2 or halogen;
R21 is hydrogen, C1- 10alkyl, C2-10alkenyl or C2-8alkynyl all of which may be unsubstituted or substituted by one or more OH, N(R5)2, CO2R5, halogen or
XC1-5alkyI or R21 is (CH2)mAr;
R22 is (CH2)m, C1- 10alkyl, C2- 10alkenyl or phenyl, all of which may be unsubstituted or substituted by one or more OH, N(R5)2, COOH, halogen, >C=O or XCμsalkyI;
R23 is hydrogen, R20, CO2R21, CO2C(R20)2O(CO)XR21, PO3(R21)2,
SO2NR21R20, NR21SO2R20, CONR21SO2R20, SO3R21, SO2R21,
P(O)(OR21)R21, CN, C(O)N(R5)2, tetrazole or OR5;
R24 is R21 or C(R16)2OC(O)R4;
m is an integer from 0 to six;
or a pharmaceutically acceptable salt thereof.
2. A compound of Claim 1 selected from the group consisting of:
(lRS)-N-[N-[1-Phosphono-3-(1,3-dioxo-1H-benz[de]isoquinoline-2-yl)propyl]-L-leucyl]-L-tryptophan
(IRS)-N-[N-[1-Phosphono-3-(1,3-dioxo-1H-benz[de]isoquinoline-2-yl)propyl]-L-leucyl]-(9-methyl-L-tyrosine
(IRS)-N-[N-[3- Amino-1-phosphono)propyl]-L-leucyl]-L-tryptophan
(IRS)-N-[N-[1-di(Methoxy)phosphinyl-3-(1,3-dioxo-1H-benz[de]isoquinoline-2-yl)propyl]-L-leucyl]-L-tryptophan
(IRS)-N-[N-[1-Phosphono-3-(1-oxo-1H-benz[de]isoquinoline-2(3H)-yl)propyl]-L-leucyl]-L-tryptophan
(IRS)-N-[N-[1-(Methoxy)phosphono-3-(1-oxo-1H-benz[de]isoquinoline-2(3H)-yl)propyl]-L-leucyl]-L-tryptophan (IRS)-N-[N-[1-Phosphono-3-N-(N-benzyloxycarbonyl-6-aminohexanamido)propyl]-L-leucyl]-L-tryptophan
3. A pharmaceutical composition comprising a compound of Claim 1 and a pharmaceutically acceptable carrier.
4. A method of inhibiting endothelin converting enzyme which comprises administering to a subject in need thereof, an effective amount to inhibit endothelin converting enzyme of a compound of Claim 1.
5. A method of treating hypertension, renal failure or cerebrovascular disease which comprises administering to a subject in need thereof, an effective amount of a compound of Claim 1.
PCT/US1994/013374 1993-11-18 1994-11-16 Endothelin converting enzyme inhibitors WO1995013817A1 (en)

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US08/154,233 1993-11-18

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US6166052A (en) * 1998-03-11 2000-12-26 Warner-Lambert Company Heteroaryl alkyl alpha substituted peptidylamine calcium channel blockers
US6251919B1 (en) 1998-02-27 2001-06-26 Warner-Lambert Heterocyclic substituted aniline calcium channel blockers
US20100158798A1 (en) * 2007-10-22 2010-06-24 Emmanuel Skordalakes FP-Pocket-Binding Effectors and Methods for Using the Same to Modulate Telomerase Activity
US9234230B2 (en) 2010-02-08 2016-01-12 The Wistar Institute Screening method and compounds for modulating telomerase activity

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US5155100A (en) * 1991-05-01 1992-10-13 Ciba-Geigy Corporation Phosphono/biaryl substituted dipeptide derivatives
US5380921A (en) * 1991-11-05 1995-01-10 Banyu Pharmaceutical Co., Ltd. Aminophosponic acid derivative
US5389612A (en) * 1989-11-15 1995-02-14 Commissariat A L'energie Atomique Phosphonic peptide inhibitors of bacterial collagenases

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US5389612A (en) * 1989-11-15 1995-02-14 Commissariat A L'energie Atomique Phosphonic peptide inhibitors of bacterial collagenases
US5155100A (en) * 1991-05-01 1992-10-13 Ciba-Geigy Corporation Phosphono/biaryl substituted dipeptide derivatives
US5380921A (en) * 1991-11-05 1995-01-10 Banyu Pharmaceutical Co., Ltd. Aminophosponic acid derivative

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6251919B1 (en) 1998-02-27 2001-06-26 Warner-Lambert Heterocyclic substituted aniline calcium channel blockers
US6166052A (en) * 1998-03-11 2000-12-26 Warner-Lambert Company Heteroaryl alkyl alpha substituted peptidylamine calcium channel blockers
US6469038B1 (en) 1998-03-11 2002-10-22 Warner-Lambert Company Heteroaryl alkyl alpha substituted peptidylamine calcium channel blockers
US6989448B2 (en) 1998-03-11 2006-01-24 Lain-Yen Hu Heteroaryl alkyl alpha substituted peptidylamine calcium channel blockers
US20100158798A1 (en) * 2007-10-22 2010-06-24 Emmanuel Skordalakes FP-Pocket-Binding Effectors and Methods for Using the Same to Modulate Telomerase Activity
US8518940B2 (en) * 2007-10-22 2013-08-27 The Wistar Institute FP-pocket-binding effectors and methods for using the same to modulate telomerase activity
US9234230B2 (en) 2010-02-08 2016-01-12 The Wistar Institute Screening method and compounds for modulating telomerase activity

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