Glaucoma treatment

Abstract: A method and a composition for treating or reducing and/or controlling intraocular pressure comprising administering an effective amount of a renin inhibiting compound of the formula: wherein A is a substituent; W is CO or CHOH and U is CH₂ or NR₂ wherein R₂ is hydrogen or loweralkyl; with the proviso that when W is CHOH then U is CH₂; R₁ is loweralkyl, cycloalkyl, benzyl, (alpha,­ alpha)-dimethylbenzyl, 4-hydroxybenzyl, 4-methoxybenzyl, halobenzyl, (1-naphthyl)methyl, (2-naphthyl)methyl, (4-imidazoyl)methyl, phenethyl, 1-benzyloxyethyl, phenoxy, thiophenoxy or anilino; R₃ is loweralkyl, loweralkenyl, ((alkoxy)alkoxy)alkyl, carboxyalkyl, (thioalkoxy)alkyl, benzyl or heterocyclic ring substituted methyl; and R₄ is a substituted amino group; or pharmaceutically acceptable salts or esters thereof. Also disclosed are compositions, methods and kits for treating glaucoma or reducing and/or controlling intraocular pressure wherein the renin inhibiting compound is administered in combination with a beta-adrenergic antagonist, a steroidal antiinflammatory agent or an angiotensin converting enzyme inhibiting compound.

Description: The present invention relates to the use of a renin inhibiting compound, separately or in combination with beta-adrenergic antagonist agents, steroidal antiinflammatory agents, or angiotensin converting enzyme inhibiting compounds, for the preparation of a topical ophthalmic composition for the treatment of glaucoma or reducing and/or controlling intraocular pressure. Background Art Glaucoma is a condition characterized by an increase in intraocular pressure. Increased intraocular pressure can lead to optic nerve damage and defects in the visual field. Blindness can result if the condition is left untreated. For some time two categories of drugs have been used to treat increased intraocular pressure. The first category is known as miotic agents and comprises cholinergic stimulating agents, such as pilocarpine, and cholinesterase antagonists, such as demacarium bromide and echothiopate iodide. Miotic agents are used to facilitate outflow of the aqueous humor. Pilocarpine has been used topically to treat glaucoma for many years and has no adverse systemic effects. However, localized adverse effects are known. Ciliary spasms may lead to debilitating myopia. Irritation and allergic reactions may require discontinuation of use. Of special concern is the known ability of miotic agents to cause retinal detachment. The cholinesterase antagonists may have systemic adverse effects including salivation, urinary incontinence, diarrhea, profuse sweating, muscle weakness, respiratory difficulties and cardiac irregularities. Localized effects include irritation, induced myopia and lens opacities. The second category of drugs used to treat increased intraocular pressure act by inhibiting the secretion of aqueous humor by the ciliary processes. Epinephrine, beta-adrenergic antagonists such as timolol and carbonic anhydrase inhibitors such as acetazolamine are members of this second category. Topical epinephrine must be used with caution in individuals with hyperthyroidism, hypertension, cardiac disease and bronchial asthma, due to possible systemic effects. Topical beta-adrenergic antagonists must also be used with caution due to a variety of systemic effects. Timolol must be used cautiously in patients with asthma, chronic obstructive pulmonary disease or sinus bradycardia. Carbonic anhydrase inhibitors are administered systemically when intraocular pressure cannot be controlled with known topical agents. Agents such as acetazolamide often cause numbness and tingling, drowsiness and loss of appetite. Recently it has been disclosed (U.S. Patent No. 4587258, E.H. Gold et al., issued May 6, 1986; U.S. Patent No. 4442089, Z.P. Horovitz, issued Apr. 10, 1984; PCT Patent Application No. WO87/02585, R.W. Watkins et al., published May 7, 1987; PCT Patent Application No. WO87/00055, R.W. Watkins et al., published Jan. 15, 1987; European Patent Application No. EP174162, G. Allan et al., published Mar. 12, 1986; European Patent Application No. EP114333, R.W. Watkins, published Aug. 1, 1984) that angiotensin converting enzyme (ACE) inhibitors are useful for treating glaucoma. When renin is released into the blood from the kidney, the renin-angiotensin system is activated, leading to vasoconstriction and conservation of sodium, both of which result in increased blood pressure. The renin acts on a circulating protein, angiotensinogen, to cleave out a fragment called angiotensin I (AI). AI itself has only slight pharmacologic activity but, after additional cleavage by a second enzyme, angiotensin converting enzyme (ACE), forms the potent molecule angiotensin II (AII). The major pharmacological effects of AII are vasoconstriction and stimulation of the adrenal cortex to release aldosterone, a hormone which causes sodium retention. AII is cleaved by an aminopeptidase to form angiotensin III (AIII), which, compared to AII, is a less potent vasoconstrictor but a more potent inducer of aldosterone release. The renin-angiotensin system has been modulated or manipulated, in the past, with ACE inhibitors to reduce blood pressure. However, ACE acts on several substrates other than angiotensin I (AI), most notably the kinins which cause such undesirable side effects as pain, "leaky" capillaries, prostaglandin release and a variety of behavioral and neurologic effects. Further, ACE inhibition leads to the accumulation of AI. Although AI has much less vasoconstrictor activity than AII, its presence may negate some of the hypotensive effects of the blockade of AII synthesis. Most recently it has been disclosed (PCT Patent Application No. WO8702581, R.W. Watkins et al., published May 7, 1987) that renin inhibitors are useful for treating glaucoma. There are no known side effects which result when renin is inhibited from acting on its substrate. While much research effort has been directed to developing renin inhibitors, there is still a need for more potent renin inhibitors. Thus, methods and compositions for treating glaucoma or reducing and/or controlling intraocular pressure which have reduced potential for localized and systemic adverse effects, as well as increased potency, are desirable. Disclosure of the Invention The present invention relates to the use of H-((beta,beta-dimethyl)-beta-Ala)-(4-0CH₃)Phe-His amide of 2(S)-amino-1-cyclohexyl-3(R),4(S)-dihydroxy-6-methylheptane which inhibits renin for the preparation of a topical ophthalmic composition for treating glaucoma or reducing and/or controlling intraocular pressure. The present invention also relates to the use of such compound in combination with a beta-adrenergic antagonist agent, an angiotensin converting enzyme inhibiting compound or a steroidal antiinflammatory agent for the preparation of a topical ophthalmic composition for treating glaucoma or reducing and/or controlling intraocular pressure. Pharmaceutical compositions disclosed herein for treating glaucoma or reducing and/or controlling intraocular pressure comprise the renin inhibiting compound in a topical ophthalmologically acceptable vehicle. The pharmaceutical compositions disclosed herein for treating glaucoma or reducing and/or controlling intraocular pressure may also comprise the above-mentioned renin inhibiting compound in combination with a beta-adrenergic antagonist agent or an angiotensin converting enzyme inhibiting compound in a topical ophthalmologically acceptable vehicle. Pharmaceutical compositions for treating the increase in intraocular pressure associated with the administration of steroidal antiinflammatory agents comprise the above-mentioned renin inhibiting compound in combination with a steroidal antiinflammatory compound in a topical ophthalmologically acceptable vehicle. The compositions may also be in the form of a kit comprising in individual containers in a single package the above-mentioned renin inhibiting compound in a topical ophthalmologically acceptable vehicle and a steroidal antiinflammatory compound in a topical ophthalmologically acceptable vehicle and/or a beta-adrenergic antagonist agent in a topical ophthalmologically acceptable vehicle or an angiotensin converting enzyme inhibiting compound in a topical ophthalmologically acceptable vehicle. The renin inhibiting compound used in the invention and a method for making it have also been disclosed in European Patent Application No. 229667, published July 22, 1987 and PCT Patent Application No. WO88/05050, published July 14, 1988. Example 1 illustrates the synthesis of H-((beta,beta-di-Me)-beta-Ala)-(4-0CH₃)Phe-His Amide of 2(S)-Amino-1-cyclohexyl-3(R),4(S)-dihydroxy-6-methylheptane Diacetic Acid Salt. Groups intended to protect the N-terminus of an amino acid or peptide or to protect an amino group against undesirable reactions during synthetic procedures or to prevent the attack of exopeptidases on the compounds or to increase the Solubility of the compounds include sulfonyl, acyl, acetyl, pivaloyl, t-butyloxycarbonyl (Boc), carbonylbenzyloxy (Cbz), benzoyl or an L- or D- aminoacyl residue, which may itself be N-protected similarly. O-protecting groups which refer to a substituent which protects hydroxyl groups against undesirable reactions during synthetic procedures include substituted methyl ethers, for example methoxymethyl, benzyloxymethyl, 2-methoxyethoxymethyl, 2-(trimethylsilyl)ethoxymethyl, benzyl and triphenylmethyl; tetrahydropyranyl ethers; substituted ethyl ethers, for example, 2,2,2-trichloroethyl and t-butyl; silyl ethers, for example, trimethylsilyl, t-butyldimethylsilyl and t-butyldiphenylsilyl; cyclic acetals and ketals, for example, methylene acetal, acetonide and benzylidene acetal; cyclic ortho esters, for example, methoxymethylene; cyclic carbonates; and cyclic boronates. The terms "Ala", "His", and "Phe", as used herein refer to alanine, histidine and phenylalanine, respectively. In general, the amino acid abbreviations follow the IUPAC-IUB Joint Commission on Biochemical Nomenclature for amino acids and peptides (Eur. J Biochem. 1984, 158 , 9-31). The terms "S" and "R" configuration are as defined by the IUPAC 1974 Recommendations for Section E, Fundamental Stereochemistry, Pure Appl. Chem. (1976) 45 , 13-30. The renin inhibiting compound used in the present invention can be used in the form of salts derived from inorganic or organic acids. These salts include the following: acetate, adipate, alginate, citrate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, camphorate, camphorsulfonate, digluconate, cyclopentanepropionate, dodecylsulfate, ethanesulfate, glucoheptonate, glycerophosphate, hemislufate, heptonate, hexanoate, fumarate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxy-ethanesulfate, lactate, maleate, methanesulfonate, nicotinate, 2-naphthaleneslufonate, oxalate, pamoate, pectinate, persulfate, 3-phenylpropionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, tosylate, and undecanoate. Also, the basic nitrogen-containing groups can be quaternized with such agents as loweralkyl halides, such as methyl, ethyl, propyl, and butyl chloride, bromides, and iodides; dialkyl sulfates like dimethyl, diethyl, dibutyl, and diamyl sulfates, long chain halides such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides, aralkyl halides like benzyl and phenethyl bromides and others. Water or oil-soluble or dispersible products are thereby obtained. Examples of acids which may be employed to form pharmaceutically acceptable acid addition salts include such inorganic acids as hydrochloric acid, sulphuric acid and phosphoric acid and such organic acids as oxalic acid, maleic acid, succinic acid and citric acid. Other salts include salts with alkali metals or alkaline earth metals, such as sodium, potassium, calcium or magnesium or with organic bases. The renin inhibiting compound used in the present invention can also be used in the form of esters. Examples of such esters include a hydroxyl-substituted compound of the invention which has been acylated with a blocked or unblocked amino acid residue, a phosphate function, or a hemisuccinate residue. The amino acid esters of particular interest are glycine and lysine; however, other amino acid residues can also be used, These esters serve as prodrugs of the compounds of the present invention and serve to increase the solubility of these substances in the gastrointestinal tract. The preparation of the pro-drug esters is carried out by reacting a hydroxyl-substituted compound of the invention with an activated amino acyl, phosphoryl or hemisuccinyl derivative. The resulting product is then deprotected to provide the desired pro-drug ester. The compositions disclosed herein are administered as topical pharmaceutical compositions suitable for ophthalmic administration, in a pharmaceutically acceptable vehicle such as pharmaceutically acceptable sterile aqueous or nonaqueous solutions, suspensions, emulsions, ointments and solid inserts. Examples of suitable pharmaceutically acceptable vehicles are water, propylene glycol and other pharmaceutically acceptable alcohols, sesame or peanut oil and other pharmaceutically acceptable vegetable oils, petroleum jelly, water soluble ophthalmologically acceptable non-toxic polymers such as methyl cellulose, carboxymethyl cellulose salts, hydroxyethyl cellulose, hydroxypropyl cellulose; acrylates such as polyacrylic acid salts; ethylacrylates; polyacrylamides; natural products such as gelatin, alginates, pectins, tragacanth, karaya, agar, acacia; starch derivatives such as starch acetate, hydroxyethyl starch ethers, hydroxypropyl starch; as well as other synthetic derivatives such as polyvinyl alcohol, polyvinyl pyrrolidone, polyvinyl methyl ether, polyethylene oxide, carbopol and xantham gum; and mixtures of these polymers. Such compositions may also contain adjuvants such as buffering, preserving, wetting, emulsifying, and dispersing agents. Suitable preserving agents include antibacterial agents such as quaternary ammonium compounds, phenylmercuric salts, benzyl alcohol, phenyl ethanol; and antioxidants such as sodium metabisulfite, butylated hydroxyanisole and butylated hydroxytoluene. Suitable buffering agents include borate, acetate, gluconate and phosphate buffers. The pharmaceutical compositions disclosed herein may also be in the form of a solid insert. A solid water soluble or water swellable polymer such as dextran, hydroxyloweralkyl dextran, carboxymethyl dextran, hydroxyloweralkyl cellulose, loweralkyl cellulose, carboxymethyl cellulose, polyvinyl alcohol, dextrin, starch, polyvinyl pyrrolidone and polyalkylene glycols may be used as the carrier for the drug. Dosage levels of the active compound in the compositions may be varied so as to obtain a desired therapeutic response to a particular composition. Generally, the active compound will be administered as an isotonic aqueous solution of from 0.00001 to 1.0 (w/v) percent concentration. More preferably the active compound will be administered as an isotonic aqueous solution of from 0.00001 to 0.1 (w/v) percent concentration. The total daily dose range for a renin inhibitor in a topical ophthalmological composition is 100 micrograms to 10 milligrams per day. The term "controlling intraocular pressure" as used herein means the regulation, attenuation and modulation of increased intraocular tension. The term also means that the decrease, in the otherwise elevated intraocular pressure, obtained by the methods and compositions described herein is maintained for a significant period of time as, for example, between consecutive doses of the composition of the invention. The renin inhibiting compound described herein may be the only active ingredient for controlling intraocular pressure in the methods and compositions described herein or may be used in combination with other ingredients which control intraocular pressure such as beta-adrenergic antagonist compounds. The term "beta-adrenergic antagonist" as used herein means a compound which by binding to beta-adrenergic plasma membrane receptors reduces or eliminates sympathetic activity or blocks the effects of exogenously adminstered catecholamines or adrenergic drugs. Examples of beta-adrenergic antagonists are atenolol, metopropol, nadolol, propranolol, timolol, labetalol, betaxolol, carteolol and dilevalol and pharmaceutically acceptable salts thereof. Most preferably the beta-adrenergic antagonist is timolol. As was previously stated, timolol is currently used for treating glaucoma or reducing and/or controlling intraocular pressure, but it has a number of adverse side effects. Accordingly, administration of a composition comprising a combination of a beta-adrenergic antagonist and the herein described renin inhibiting compound could produce a reduction in intraocular pressure equivalent to that produced by a beta-adrenergic antagonist alone, but at a reduced dose level of the beta-adrenergic antagonist. This will result in a reduced level of the beta-adrenergic antagonist related adverse side effects. The combination composition is administered as a single dosage form containing both the herein described renin inhibitor and the beta-adrenergic antagonist. The beta adrenergic antagonist may comprise from 5 mg to 125 mg of the composition of the invention. The preferred ranges of the components in the composition of the invention in unit dosage form are: Renin inhibitor: 1 ng to 0.250 mg Beta-adrenergic antagonist: 5 mg to 125 mg When the beta-adrenergic antagonist and the renin inhibitor are administered as separate compositions the composition is in the form of a kit comprising in two separate containers a topical ophthalmologically acceptable beta-adrenergic antagonist composition and a topical ophthalmologically acceptable renin inhibitor composition, in a single package. The renin inhibiting compound may also be administered in combination with an angiotensin converting enzyme inhibiting compound. Examples of angiotensin converting enzyme inhibiting compounds are captopril and enalapril. As was previously mentioned, ACE inhibitors have some undesirable side effects, Accordingly, administration of an ACE inhibitor in combination with a renin inhibitor could produce a reduction in intraocular pressure greater than or equivalent to that of an ACE inhibitor alone, but at a reduced dose level of the ACE inhibitor. This will result in a reduced level of the ACE inhibitor related adverse side effects. The combination composition is administered as a single dose form containing both the renin inhibitor and the angiotensin converting enzyme inhibitor. The ACE inhibitor may comprise from 5 ng to 50 micrograms of the composition. The preferred ranges of the components in the composition in unit dosage form are: Renin inhibitor: 1 ng to 0.250 mg ACE inhibitor: 5 ng to 50 micrograms When the ACE inhibitor and the renin inhibitor are administered as separate compositions the composition is in the form of a kit comprising in two separate containers a topical ophthalmologically acceptable ACE inhibitor composition and a topical ophthalmologically acceptable renin inhibitor composition, in a single package. Dosage levels of the active compounds in the compositions disclosed herein may be varied so as to obtain a desired therapeutic response depending on the route of administration, severity of the disease and the response of the patient. Administration of steroidal antiinflammatory agents can cause an increase in intraocular pressure. The increase in intraocular pressure can be reduced by the administration of the renin inhibiting compound described herein. Steroidal antiinflammatory agents include hydrocortisone, cortisone, prednisone, prednisolone, dexamethasone, methylprednisolone, triamcinolone, betamethasone, alclometasone, flunisolide, beclomethasone, clorocortolone, diflorasone, halcinonide, fluocinonide, fluocinolone, desoximetasone, medrysone, paramethasone, and fluorometholone, and their pharmaceutically acceptable salts and esters. Preferred steroidal antiinflammatory agents are hydrocortisone, prednisolone, dexamethasone, medrysone and fluorometholone and their pharmaceutically acceptable salts and esters. The renin inhibitor is administered after use of a steroidal antiinflammatory agent or at the same time, causing reduction and/or control of intraocular pressure. When the combination of a steroidal antiinflammatory agent and the renin inhibitor is used, a preferred combination is a topical ophthalmic dosage form comprising both a steroidal antiinflammatory and the renin inhibitor described herein. When the steroidal antiinflammatory agent and the renin inhibitor are administered as separate compositions the composition is in the form of a kit comprising in two separate containers a topical ophthalmologically acceptable steroidal antiinflammatory agent composition and a topical ophthalmologically acceptable renin inhibitor composition, in a single package. The combination composition disclosed herein may contain from 0.00001 to 1.0 (w/v) percent of the renin inhibitor for combined or separate topical administration. More preferably the amount of the renin inhibitor is 0.00001 to 0.1 (w/v) percent of the composition. The amount of the renin inhibitor in a unit dosage form for topical administration to the eye is from 5 ng to 0.5 mg, preferably from 5 ng to 250 micrograms. The dose required will depend on the potency of the particular renin inhibitor, the severity of the intraocular pressure increase and the response of the individual patient. The combination composition disclosed herein may contain from 0.05 to 1.5 (w/v) percent of the steroidal antiinflammatory for combined or separate topical administration. The amount of the steroidal antiinflammatory in a unit dosage form for topical administration to the eye is from 20 micrograms to 600 micrograms. The dose required will depend on the potency of the particular steroidal antiinflammatory, the severity of the disease and the response of the individual patient. The compositions disclosed herein may include other therapeutic agents in addition to the renin inhibitor, and other agents which reduce and/or control intraocular pressure. In the test data following Example 1, H-((beta,beta-dimethyl)-beta-Ala)-(4-OCH₃)Phe-His amide of 2(S)-amino-1-cyclohexyl-3(R),4(S)-dihydroxy-6-methylheptane is referred to as Compound V. Example 1 describes procedures useful for the preparation of H-((beta,beta-di-Me)-beta-Ala)-(4-0CH₃)Phe-His Amide of 2(S)-Amino-1-cyclohexyl-3(R),4(S)-dihydroxy-6-methylheptane Diacetic Acid Salt. Example 1 a. Boc-Phe-Ala Amide of (2S)-Amino-1-cyclohexylbut-3-ene 2(S)-t-Butyloxycarbonylamino-1-cyclohexylbut-3-ene (310 mg, 1.22 mmol) was dissolved in 1 M anhydrous HCl in anhydrous methanol (35 mL). After 22 hours, the solvent was evaporated to give 230 mg (99%) of the corresponding amine hydrochloride which was used without further purification. To a stirred -13°C solution of Boc-Phe-Ala (408 mg, 1.21 mmol) in dry THF (8 mL) containing N-methylmorpholine (122 mg, 1.21 mmol) was added isobutyl chloroformate (165 mg, 1.21 mmol) dropwise. After 3 minutes, a -13°C solution of the above amine hydrochloride (230 mg, 1.21 mmol) in 1:1, THF:dimethyl formamide (DMF) (4 mL) containing N-methylmorpholine (122 mg) was added dropwise. The mixture was warmed to room temperature for 2 hours. Evaporation provided a residue which was partitioned between ethyl acetate (30 mL) and 0.1 M H₃PO₄ (10 mL). The organic phase was washed with brine (10 mL), saturated NaHCO₃ (10 mL), and brine (10 mL). Drying, filtering, evaporating, and chromatographing (55 g SiO₂; 95:5, CH₂Cl₂:CH₃OH) gave the desired compound (462 mg, 81%). b. Boc-Phe-His Amides of 3(S)-Amino-4-cyclohexyl-2(R,S)-hydroxy-1-t-butyldimethylsilyloxybutane 3(S)-t-Butyloxycarbonylamino-4-cyclohexyl-1,2(R,S)-dihydroxybutane (1.10 g, 3.82 mmol) was treated with anhydrous 1M HCl/CH₃OH (80 mL) for 16 hours at which time evaporation and drying provided the corresponding amine hydrochloride (0.85 g, 100%). To a suspension of the above hydrochloride salt (344 mg, 1.54 mmol) and imidazole (105 mg) in dichloromethane (15 mL) were added triethylamine (156 mg) and t-butyldimethylsilyl chloride (232 mg). The solvent was evaporated after 31 hours, and the residue was then re-dissolved in anhydrous dimethylformamide (DMF, 15 mL). Boc-Phe-His (619 mg) and 1-hydroxybenzotriazole (HOBT, 312 mg) were then added. After cooling the stirred solution to -23°C, 1,3-dicyclohexylcarbodiimide (DCC, 318 mg) was added. The mixture was warmed to room temperature 3 hours later. After 13 hours the solvent was evaporated in vacuo, and the residue was dissolved in ethyl acetate (40 mL), filtered, washed with saturated NaHCO₃ (2x10 mL) and brine (10 mL), and dried (Na₂SO₄). Filtration and evaporation provided a residue which was chromatographed on silica gel eluting with dichloromethane/methanol mixtures to give 441 mg (42%) of the desired product. Mass spectrum: (M+H)⁺ = 686. Anal. calcd. for C₃₆H₅₉N₅O₆Si: C, 63.0; H, 8.7; N, 10.2. Found: C, 62.8; H, 9.0; N, 9.9. c. 2(S)-t-Butyloxycarbonylamino-1-cyclohexyl-6-methylhept-3-ene To a stirred -78°C solution of Boc-cyclohexylalanine methyl ester (40 g, 140 mmol) in anhydrous toluene (250 mL) was added diisobutylaluminum hydride (130 Mol%, 1.5 M solution in toluene, 121.4 mL) at a rate to keep the internal temperature below -60°C. After stirring for an additional 20 minutes at -78°C, the aldehyde solution is used immediately as described below. To a potassium hydride (35% dispersion in oil, 32.09 g) suspension in a 0°C mixture of anhydrous THF/DMSO (1000 mL/200 mL) under dry N₂ was added 1,1,1,3,3,3-hexamethyldisilazane (209 Mol%, 49.07 g) dropwise. After stirring at 0°C for 1 hour, the resulting solution was added via cannula to a 0°C flask containing isopentyltriphenylphosphonium bromide (209 Mol%, 125.66 g). The mixture was stirred vigorously for 1 hour at which time it was cooled to -78°C. The -78°C aldehyde solution prepared above was then added via cannula. After stirring at -78°C for 15 minutes, the mixture was allowed to slowly warm to room temperature and then heated to 40°C for 12 hours. The mixture was then cooled to room temperature and quenched with methanol (7.65 mL) followed by aqueous Rochelle salts (100 mL saturated solution and 500 mL H₂O). The mixture was then extracted with ethyl acetate (2x). The combined extracts were washed with water and brine. Drying (MgSO₄) and evaporating provided crude alkene which was chromatographed on silica gel (ether/hexane) to give 16.5 g (38%) of the desired compound as an 85:15 mixture of cis:trans isomers. Mp=53-55°C. Mass spectrum: M⁺ = 309. Anal. calcd. for C₁₉H₃₅NO₂: C, 73.7; H, 11.4; N, 4.5. Found: C, 73.8; H, 11.4; N, 4.5. d. 2(S)-t-Butyloxycarbonylamino-1-cyclohexyl-3,4-dihydroxy-6-methylheptane: The 3(R)4(S), 3(S)4(S), 3(R)4(R), and 3(S)4(R) Diastereomers To a solution of the resultant compound of Example 5c (8.50, 27.5 mmol) in dry THF (150 mL) were added OsO₄ (2.8 mL of a 2.5% solution in t-butanol and N-methylmorpholine N-oxide (9.28 g, 68.7 mmol). After 4 days the mixture was partitioned between ether (200 mL) and brine (100 mL). The aqueous layer was back-extracted with ether (2 x 100 mL), and the combined organic phase was washed with 10% Na₂SO₃, 0.1 M H₃PO₄, and brine. Drying (MgSO₄) and evaporating provided a residue (10.81 g) which was chromatographed on silica gel to elute a 60% yield of the 4 diols in the following order. 3(R),4(S) Mass spectrum: (M+H)⁺ = 344. Anal. calcd. for C₁₉H₃₇NO₄: C, 66.4; H, 10.9; N, 4.1. Found: C, 66.4; H, 10.8; N, 3.9. 3(S),4(S) Mass spectrum: (M+H)⁺ = 344. Anal. calcd. for C₁₉H₃₇NO₄: C, 66.4; H, 10.9; N, 4.1. Found: C, 66.4; H, 11.1; N, 4.0. 3(R),4(R) Mass spectrum: (M+H)⁺ = 344. 3(R),4(R) Mass spectrum: (M+H)⁺ = 344. Anal. calcd. for C₁₉H₃₇NO₄: C, 66.4; H, 10.9; N, 4.1. Found: C, 66.0; H, 10.7; N, 4.0. e. Boc-Phe-His Amide of 2(S)-Amino-1-cyclohexyl-3(R),4(S)-dihydroxy-6-methylheptane The 3(R),4(S) diastereomer of Example 5d was deprotected with HCl/methanol, and the resulting product was coupled to Boc-Phe-His using 1-hydroxybenzotriazole and 1,3-dicyclohexylcarbodiimide according to the procedure of Example 5b. The desired product was obtained in 40-60% yield, Mass spectrum: (M+H)⁺ = 628. Anal. calcd. for C₃₄H₅₃N₅O₆ H₂O: C, 63.2; H, 8.6; N, 10.8. Found: C, 63.2; H, 8.4; N, 10.5. f. Boc-His Amide of 2(S)-Amino-1-cyclohexyl-3(R),4(S)-dihydroxy-6-methylheptane The procedure of Example 5e was followed except Boc-Phe-His was replaced with Boc-His. Mass spectrum: (M)⁺ = 480. Anal. calcd. for C₂₅H₄₄N₄O₅ 3/4H₂O: C, 60.8; H, 9.1; N, 11.3. Found: C, 60.9; H, 9.2; N, 11.0. g. TBA-CHA-His Amide of 2(S)-Amino-1-cyclohexyl-3(R),4(S)-dihydroxy-6-methylheptane The resultant compound of Example 5f was deprotected with HCl/methanol, and the resulting product was coupled to t-butylacetyl-cyclohexylalanine (TBA-CHA) using the DCC/HOBT method of Example 5b. High resolution mass spectrum calcd. for C₃₅H₆₁N₅O₅, (M+H) 632.4751. Found: 632.4759. h. Cbz- beta -Ala-Phe-His Amide of 2(S)-Amino-1-cyclohexyl-3(R),4(S)-dihydroxy-6-methylheptane Using the procedure of Example 5g, but replacing TBA-CHA with Cbz-beta-Ala-Phe gave the desired compound. Mass spectrum: (M+H)⁺ = 733. Anal. calcd. for C₄₀H₅₆N₆O₇: C, 65.5; H, 7.7; N, 11.5. Found: C, 65.2; H, 7.7; N, 11.2. i. H- beta -Ala-Phe-His Amide of 2(S)-Amino-1-cyclohexyl-3(R),4(S)-dihydroxy-6-methylheptane Diacetic Acid Salt The resultant compound of Example 5h (1.00 g, 1.36 mmol) in acetic acid (14 mL) was hydrogenated at 1 atmosphere with 10% Pd/C (0.50 g) for 3 h. Filtration, extraction of the catalyst with acetic acid, and evaporation of the combined acetic acid solutions gave a residue which was dissolved in water (25 mL) and lyophilized to provide 891 mg (91%) of the desired product. Mass spectrum: (M+H)⁺ = 599 (free base). Anal. calcd. for C₃₆H₅₈N₆O₉ 1/2 H₂O): C, 59.4; H, 8.1; N, 11.5. Found: C, 59.3; H, 8.0; N, 11.2. j. Cbz- ( (beta,beta-di-Me)-beta-Ala)-Phe-His Amide of 2(S)-Amino-1-cyclohexyl-3(R),4(S)-dihydroxy-6-methylheptane Using the procedure of Example 5g, but replacing TBA-CHA with Cbz-((beta,beta-di-Me)-beta-Ala)-Phe gave the desired compound. Mass spectrum: (M+H)⁺ = 761. Anal. calcd. for C₄₂H₆₀N₆O₇ 1/4H₂O: C, 65.5; H, 8.0; N, 10.9. Found: C, 65.5; H, 7.9; N, 11.0. k. H- ( (beta,beta-di-Me)-beta-Ala)-Phe-His Amide of 2(S)-Amino-1-cyclohexyl-3(R),4(S)-dihydroxy-6-methylheptane Diacetic Acid Salt Using the procedure of Example 5i, but replacing the resultant compound of Example 5h with the resultant compound of Example 5j gave the desired product. Mass spectrum: (M+H)⁺ = 627 (free base). Anal. calcd. for C₃₈H₆₂N₆O₉.H₂O: C, 59.7; H, 8.4; N, 11.0. Found: C, 59.5; H, 8.4; N, 11.3. l. H- ( (beta,beta-di-Me)-beta-Ala)-(4-OCH₃)Phe-His Amide of 2(S)-Amino-1-cyclohexyl-3(R),4(S)-dihydroxy-6-methylheptane Diacetic Acid Salt Using the procedures of Examples 5j and 5k, but replacing Cbz-((beta,beta-di-Me)-beta-Ala)-Phe with Cbz-((beta,beta-di-Me)-beta-Ala)-(OCH₃)Phe gave the desired product. (M+H)⁺ = 657 (free base). Effects of Topically Administered Renin Inhibiting Compound V on Intraocular Pressure of Rabbits Method The antiglaucoma activity of the compounds was tested by measuring the effect on intraocular pressure in rabbits as described by Tinjum, A.M., Acta Ophthalmologica, 50 , 677 (1972). Male albino, New Zealand rabbits were placed in restraining devices and the intraocular pressure was measured with an applamatic tonometer. Exactly 0.1 ml of an isotonic saline solution containing the test compound was instilled into the conjuctival sac and the intraocular pressure was measured at 5, 15, 30, 60, 90, 120 and 150 minutes afterwards. The results obtained show that Compound V is able to reduce intraocular pressure in vivo . Dose Response Study of the Effects of Topically Administered Renin Inhibiting Compound V on Intraocular Pressure of Rabbits a. Method Using the method cited above, various topically administered doses of Compound V and timolol were studied. b. Results Mean Intraocular Pressure in mmHg Minutes after Administration Treatment CONTROL 5 15 30 60 90 120 150 Vehicle 20.1 19.1 21.5 19.2 20.7 20.7 18.8 21.3 Compound V 0.01% 19.2 17.7 16.3 13.7 16.9 15.4 18.2 20.3 Compound V 0.03% 19.0 13.8 13.3 12.8 13.8 13.8 15.2 16.8 Compound V 0.1% 18.7 14.1 13.1 11.9 11.3 11.5 17.5 19.0 Compound V 0.3% 21.2 13.3 12.0 10.9 12.2 12.4 16.0 18.8 Timolol 0.02% 19.1 16.5 16.6 13.9 19.1 18.9 22.4 19.9 Timolol 0.05% 18.2 15.4 12.8 12.8 12.3 15.6 17.8 20.3 Timolol 0.2% 18.6 12.1 12.5 14.7 12.8 15.6 15.9 18.9 Timolol 0.5% 18.4 12.8 12.8 11.0 10.9 16.1 17.2 18.7 Each value cited represents the average of 6 observations. The contralateral eye in each rabbit was untreated. The results indicate that the intraocular pressure lowering effect of Compound V lasts 30 to 60 minutes longer than timolol at a dose one half that of timolol. Effect of Topically Administered Compound V Alone and In Combination With Timolol On Intraocular Pressure of Rabbits a. Method Using the method cited above, topically administered doses of Compound V and timolol, alone and in combination, were studied. b. Results Mean Intraocular Pressure in mmHg Minutes after Administration Treatment CONTROL 5 15 30 60 90 120 150 Vehicle 20.1 19.1 21.5 19.2 20.7 20.7 18.8 21.3 Compound V 0.03% 17.5 10.4 10.3 10.1 10.2 10.4 14.5 17.0 Timolol 0.05% 16.8 10.2 11.0 10.1 9.9 13.9 14.5 17.0 Compound V 0.03% and Timolol 0.05% 18.5 10.9 10.5 9.5 11.6 14.0 14.7 15.3 Each value cited represents the average of 6 observations. The contralateral eye in each rabbit was untreated. The results indicate that the intraocular pressure lowering effect of a combination of Compound V and timolol has a longer duration of action than either compound when administered alone.

Claims: The use of H-((beta,beta-dimethyl)-beta-Ala)-(4-OCH₃)Phe-His amide of 2(S)-amino-1-cyclohexyl-3(R),4(S)-dihydroxy-6-methylheptane or a pharmaceutically acceptable salt or ester thereof for the preparation of a topical ophthalmic composition for treating glaucoma or controlling intraocular pressure. The use of H-((beta,beta-dimethyl)-beta-Ala)-(4-OCH₃)Phe-His amide of 2(S)-amino-1-cyclohexyl-3(R),4(S)-dihydroxy-6-methylheptane or a pharmaceutically acceptable salt or ester thereof, in combination with a beta-adrenergic antagonist compound or pharmaceutically acceptable salt thereof, for the preparation of a topical ophthalmic composition for treating glaucoma or controlling intraocular pressure. The use of H-((beta,beta-dimethyl)-beta-Ala)-(4-OCH₃)Phe-His amide of 2(S)-amino-1-cyclohexyl-3(R),4(S)-dihydroxy-6-methylheptane or a pharmaceutically acceptable salt or ester thereof, in combination with an angiotensin converting enzyme inhibiting compound, for the preparation of a topical ophthalmic composition for treating glaucoma or controlling intraocular pressure. The use of H-((beta,beta-dimethyl)-beta-Ala)-(4-OCH₃)Phe-His amide of 2(S)-amino-1-cyclohexyl-3(R),4(S)-dihydroxy-6-methylheptane or a pharmaceutically acceptable salt or ester thereof, in combination with a steroidal antiinflammatory agent or a pharmaceutically acceptable salt or ester thereof, for the preparation of a topical ophthalmic composition for treating glaucoma or controlling intraocular pressure.

Inventors: CROWLEY STEVEN R, PLATTNER JACOB J, STEIN HERMAN H, CROWLEY, STEVEN R., PLATTNER, JACOB J., STEIN, HERMAN H.
Applicants: ABBOTT LAB, ABBOTT LABORATORIES
Application year: 1988
Main classification: A61K38
IPC codes: C07D295, A61K31, C07D211, C07C317, C07C271, C07C305, A61P43, C07D413, C07C323, C07D233, C07C237, C07D405, C07C235, A61K38, C07D263, C07C309, C07C233, A61P27, C07C307