NZ196764A

NZ196764A - Hydroxynaphthoic acid ureides

Info

Publication number: NZ196764A
Application number: NZ19676481A
Authority: NZ
Inventors: R B Conrow; S Bernstein
Original assignee: American Cyanamid Co
Priority date: 1981-04-08
Filing date: 1981-04-08
Publication date: 1984-03-16

Description

<div class="application article clearfix" id="description"> New Zealand Paient Spedficaiion for Paient Number 1 96764 1 /7 Priority 8- H- ■ 81 Compters GpscifScetJon Fited: hume WMM; ■ Pyjffilfl; m' : * • )Si5 S Publication Data: P 0. Joumm, wo: t--u—<ri£±* i a i i I • PI '^r ^ ^ f| f 114 < tioj yi i $%} NEW ZEALAND PATENTS ACT, 1953 it- No.: Date: v COMPLETE SPECIFICATION SUBSTITUTED NAPHTHOIC ACIDS X/We, AMERICAN CYANAMID COMPANY, a corporation organized and existing under the laws of the State of Maine, United States of America, and having its executive cff ices at Wayne, New Jersey, United States gf America, hereby declare the invention for which X / we pray that a patent may be granted to flsus/us, and the method by which it is to be performed, to be particularly described in and by the following statement:- - 1 - (followed by la) 1 - la- 5 SUBSTITUTED NAPHTHOIC ACIDS 10 This invention relates to novel C-substituted naph thoic acid ureides which may be represented by Formula I: 15 20 25 30 HCO NHCO 12 ~ (I) wherein A is selected from the group consisting of hydrogen, lower (C]_-C(j) alkyl and a pharmaceutical^ acceptable salt cation; B is selected from the group consisting of hydrogen, lower (C^-Cg) alkanoyl and a pharmaceutically acceptable salt cation; and R is selected from the group consisting of hydrogen and lower (C1-C3) alkyl. These compounds are useful as inhibitors of connective tissue destruction. Abnormal destruction of connective tissue by colla-genase and/or neutral proteases causes tissue damage and/or tissue dysfunction. In these conditions an inhibitor of connective tissue destruction acting directly or indirectly would be useful in preventing, retarding, or reversing tissue damage and/or collagen diseases. The term connective tissue refers to a matrix of at least three protein molecules, collagen, proteoglycan and - 2 - elastin. These molecules play an important role in the structural integrity of normal tissues. Collagen, the most abundant protein in the body occupies a central position in the connective tissue matrix ["Biochemistry of Collagen", Ed. G. N. Ramachandran and A. H. Reddi, Academic Press, New York (1976); P. Bornstein, Ann. Rev. Biochem., 43, 567 (1974); J. Fessler and L. Fessler, Ann. Rev. Biochem., 47, 129 (1978)]. Collagen is, for example, the main structural component of the oral tissue (periodontal, ligament, alveolar bone, gingiva, and cementum) [Fullmer, e_t al_. , J. Dental Research, 48, 646 (1969)]. collagen amounts to 40% of cartilage protein, 90% of bone protein, and over 90% of dry dermis. Articular cartilage is the resilient tissue that covers the articulating extremities in synovial joints. It consists of collagen fibres that are intimately meshed in a hydrated gel of proteoglycan. Proteoglycan, as it exists in cartilage, is a molecule in which sulfated polysaccharide chains are covalently linked to a protein backbone ["Dynamics of Connective Tissue Macromolecules", Ed. P. M. burleigh and A. R. Poole, North Holland, Amsterdam (1975)]. Elastin is a major connective tissue component of pulmonary structure ["Elastin and Elastic Tissue", Ed. L. B. Sandberg, W. R. Gray, and C. Franzblau, Plenum Press, New York (1977)]. The breakdown of elastin of pulmonary connective tissue is considered the primary event in pulmonary emphysema [A. Janoff in "Proteases and Biological Control", Cold Spring Harbor Conference on Cell Proliferation, 2, 603 (1975)]. Degradation of fibrous collagern is initiated by a combination of neutral proteases and tissue collagenase as an integral part of a complex immunopathological process which results in the loss of collagen from normal tissue. Under normal conditions cellular mechanisms maintain a careful balance between the rates of collagen synthesis and degradation. However, in certain pathological conditions, the ensuing i 967 64 - 3 - elevated levels of neutral proteases and collagenase can result in rapid collagen degradation and tissue dysfunction. For example, in periodontal disease, the generated elevated levels of neutral proteases and collagenase in the gingival crevicular fluid rapidly degrade the fibrous collagen suppor-ing the teeth. Periodontal pockets result ultimately from collagen degradation and as these pockets deepen, support of tooth is lost and alveolar bone is resorbed [K. Ohlsson, I. Ohlsson, and G. I. Basthall, Acta Odontol. Scand., 32, 51 (1974); L. M. golub, S. Kenneth, H. McEwan, J. B. Curran, and N. S. Ramamurthy, J. Dental Research, 55, 177 (1976); L. M. Golub, J. E. Stakin and D. L. Singer, J. Dental Research, 53, 1501 (1974); -L. M. Wahl, S. M. Wahl, S. E. Mer-genhagen, and G. R. Martin, Proc. Natl. Acad. Sci. U.S., 71, 3598 (1974); Science, 187, 261 (1975)]. In arthritic conditions such as in rheumatoid arthritis septic arthritis, and osteoarthritis elevated degradation of.collkagen and proteoglycan initiate rapid destruction of articular tissue [J. M. Evanson, J. J. Jefferey, and S. M. Krane, Science, 158, 499 (1967); E. D. Harris, D. R. Dibona and S. M. Krane, J. Clin. Invest., 48, 2104 (1969); E. D. Harris, Rheumatoid Arthritis, Medcom. Press, N. Y. (1974); Z. Werb, C. L. Mainardi, C. A. Vater and E. D. Harris, New Eng. J. Med., 296, 1017 (1977); J. M. Dayer, R. G. Russell and S. M. Krane, Science, 195, 181 (1977); E. D. Harris, C. A. Vater, C. L. Mainardi and Z. Werb, Agents and Actions, 8, 35 (1978); D. E. woolley, E. D. Harris, C. L. Mainardi and C. E. Brinkerhoff, Science, 200, 773 (1978); E. D. Harris, C. S. faulkner, F. E. Brosn, Clin. Orthoped., 110, 303 (1975); M. G. Ehrlich, H. J. Mankin, H. Jones, R. Wright and C. Crispe J. Bone Jt. Surg., 57A, 565 (1975); S. gordon, W. Newman and B. Bloom, Agents and Action, 8_, 19 (1978); "Mechanisms of Tissue Injury With Reference to Rheumatoid Arthritis", Ed. R. J. Perper, Ann. N. Y. Acad. Sci., 256, 1-450 (1975)]. Increasec collagen degradation in bone can result in abnormal bone destruction as in osteoporosis [C. G. Griffith, Nichols, J. D. Asher and B. Flannagan, J. Am. Med. Assoc., 193, 91 (1965); 3. Gardner, H. Gray and G. Hedyati, Curr. Top Surg. Res., 2, 175 (1970); B. Gardner, S. Wallach, H. Gray and R. K. Baker, Surg. Forum, 22, 435 (1971)]. Collagenase activity has also resulted in tissue damage in cholesteatoma [M. Abramson, R. W. S. Schilling, C. C. Huang and R. G. Salome, Ann. Otol. Rhinol. Faryngol., 81, 158 (1975); M. Abramson and C. C. Huang, Laryngoscope, 77, 1 (1976)1. In corneal ulcerations that progress to loss of corneal integrity and function, collagenase has been implicated as a direct factor in corneal destruction [S. I. Brown, C. W. Hook and N. P. Tragakis, Invest. Ophthamol. , 1_1, 149 (1972); M. B. Berman, C. H. Dohlman, P.F. Davison, and M. Ghadringer, Exptl. Eye Res., 11, 225 (1971)]. Elevated levels of collagenase have also been observed in patients with epidermo-lysis bullosa,, and a group of related genetic diseases of the skin [E. A. Bauer, T. G. Dahl, and A. Z. Eisen, J. Invest. Dermatology, 68, 119 (1977) . Increased breakdown of elastin of the lung tissue by neutral proteases (elastase) may contribute to the lesions in pulmonary emphysema [I. Mandel, T. V. Darmle, J. A. Frier-er, S. Keller and G. M. Turino, Elastin and Elastic Tissue, Ed. L. B. Sandberg, W. R. Gray and C. Fransblau, Plenum Press N. Y., p. 221 (1977) ] . A variety of substances, both .naturally occurring and synthetically prepared, have been found to be inhibitors of connective tissue destruction, e.g., inhibitors of collagen degradation, that is, as collagenase inhibitors. Such substances include, for example, ethylenediaminetetraacetate, 1,10-phenanthroline, cysteine, dithiothretol and sodium aurio thiomalate [D. E. Woolley, R. W. glanville, D. R. Roberts and J. M. Evanson, Biochem J., 169 265 (1978); S. Seifter and E. Harper, Chap. 18, The Collagenases" in The Enzymes (3rd Edition), 3_, 649-697, Ed. by P. D. Boyer, Academic Press, N.Y (1971)]. In the eye, a number of studies using collagenase inhibitors directly applied to corneal ulcerations have been reported. Calcium ethylenediaminetetraacetate and acetylcysteine reduce the frequency of ulceration in the alkali 196764 - 5 - burned rabbit [M. Berman and C. Dohlman, Arch. Ophthamol., 35, 95 (1975)]. Both cysteine and acetylcysteine have been effective in the treatment of acute and chronic corneal ulceration in the human, although the latter compound was preferred because of its greater stability [S. I. Brown, N. P. Tragakis and D. B. Pease, Am. J. Ophthamol., 7_4, 316 (1972); M. Berman, Trace Components of Plasma: Isolation and Clinical Significance, 7th Annual Red Cross Symposium, p. 225, Alan R. Liss, Inc., N. Y. (1976)]. Naturally .occurring collagenase inhibitors include the serum components alpha2~macroglobulin and beta^-anti-collagenase [D. E. Woolley, R. W. glanville, D. R. Roberts and J. M. Evanson, Biochem. J., 169, 265 (1978)]. While some compounds may inhibit the destructive effect of collagenase on connective tissue by acting directly on collagenase itself, other compounds may inhibit such destruction by coating, binding or competing with sites on the connective tissue in such a manner as to prevent collagenase from attacking it. The present invention, however, is not to be restricted or limited to any particular mechanism or mode of action. Suffice it to say, that the compounds of this invention have utility as inhibitors of connective tissue destruction- albeit in whatever manner or \ mode. ♦ U. S. Patent No. 2,687,436 discloses substituted 3-(2-naphthyl)-cyclohexanes juseful in the treatment of collagen diseases. British Patent Nos. 856,357 and 1,246,141 disclose 2-aryl-hexahydro-quinolizines and l-hydroxyproline derivatives, respectively, useful for treating diseases affecting connective tissue. The closest known structurally related compound to those of the present invention and disclosed as having collagenase inhibiting activity is found in Thromb. Res., 10(4), 605-11 (1977), wherein the trypanocidal agent trypan blue is reported as inhibiting the activity of collagenase, or a. proteinase contaminant in the collagenase preparation. It is 'interesting, however, that in this same ticle, the ureide Suramin is reported as not inhibiting the action of collagenase. The closest known ureides to those of the present invention, and not disclosed as inhibi tors of connective tissue destruction or as collagenase inh bitors are those ureides found in Journal of the Chemical], Society, 3069 (1927). Of particular interest are the group of compounds encompassed within Formula I and illustrated by Formulas II and III: NHCO (II) (III) 5 10 i96764 - 7 - wherein A, B and R are as defined with reference to Formula I. By pharmaceutical^ acceptable salt cation is meant an alkali metal; and alkaline earth metal; ammonium; primary amine, e.g. ethyl amine; secondary amine, e.g. di-ethylamine or diethanolamine; tertiary amine, e.g. pyridine, triethylamine or 2-dimethylaminomethyldibenzofuran; aliphatic diamine, e.g. decamethylenediairiine; or an aromatic diamine. Representative compounds encompassed within this invention include, for example: 6,6'-[Ureylenebis(m-phenylenecarbonylimino)]bis[4-hydroxy-2--naphthoic acid] diethyl ester diacetate 6,6'-[Ureylenebis( m-phenylenecarbonylimino)]bis[4-hydroxy-2--naphthoic acid 6,6'-[Ureylenebis Cm-phenylenecarbony1imino)]bis[4-hydroxy-2--naphthoic acid] diethyl ester This invention is also concerned with a method of inhibiting connective tissue destruction in a warm-blooded animal which comprises administering to said animal an effective inhibiting amount of a compound encompassed within Formula I. Moreover, this invention is concerned with a method of inhibiting the degradation sequelae of collagenase activity in a body fluid, such as crevicular fluid, synovial fluid and the like, which comprises subjecting body fluid collagenase to the action of an effective collagenase inhibiting amount of a compound encompassed- within the above formula. i?APRl983 j Body fluid can include blood, plasma, serum, synovial fluid crevicular fluid, ocular fluid, etc., containing collagenase. The method of use aspect of this invention is further concerned with a method of inhibiting the action of collagenase in a warm-blooded animal which comprises internally administering to said animal an effective collagenase inhibiting amount of a compound encompassed within the above formula. find utility as inhibitors of connective tissue destruction or as collagenase inhibitors in body fluids, as such they may be useful in ameliorating or preventing those pathological reactions resulting from the functioning of collagenase, and in the therapeutic treatment of warm-blooded animals having connective tissue disorders such as periodontal diseases and diseases of the teeth, osteoporosis, Paget's disease, hyperparathyroidism or renal failure, rheumatoid arthritis, septic arthritis, osteoarthritis, gout, acute synovitis, scleroderma, psoriasis, epidermolysis bullosa, keloids, blisters, cholesteatoma of the ear, and corneal ulceration. The compounds of the present invention may also be useful in those pathological states where excessive activity of neutral proteases causes tissue damage. The compounds of the present invention may be pre pared according to the following Flowchart A. Since the compounds of the present invention 2 7 APR 1983 4 Mr Flowchart A 1 sr\_ y, COCl NO- NO. NHCO -—^ ^ ^ N ri 2 NHCO R 2 CO 1 cr^ ^ It? - - With reference to Flowchart A, a substituted--amino-naphthoic acid 1_ is dissolved in pyridine, cooled and reacted with an excess substituted nitrobenzoylchloride 2, giving a substituted nitrobenzamido-substituted naph-5 thoic acid 2> which is hydrogenated in the presence of a suitable catalyst to give the corresponding amine derivative 4. The amine 4 is dissolved in pyridine and phosgenated to give the final ureide product 5_ which is isolated by conventional procedures. 10 15 20 25 30 196764 - ii - Example 1 6-(m-Aminobenzamido)-4-hydroxy- -2-naphthoic acid, ethyl ester, acetate A solution of 20 g. of 4-hydroxy-6-nitro-2-naph-5 thoic acid [W. F. Beech and N. Legg, J. Chem. Soc., 1887 (1949)], 385 ml. of absolute ethanol and 20 ml. of concentrated sulfuric acid is refluxed for 4 hours, concentra-ated and diluted with water. The solid is collected by filtration, washed with water until neutral and crystallized 10 from 250 ml. of acetonitrile, giving 16.5 g. of 4-hydroxy--6-nitro-2-naphthoic acid ethyl ester as yellow crystals. To a mixture of 22.47 g. of 4-hydroxy-6-nitro-2--naphthoic acid ethyl ester in 150 ml. of pyridine is added 8.5 ml. of acetic anhydride. The mixture is stirred for 5 15 minutes, warmed on a steam bath until solution is complete and then allowed to stand for 10 minutes. The solution is poured into one liter of ice water and then filtered. The solid is dissolved in 500 ml. of methylene chloride, dried over sodium sulfate, filtered and concentrated to about 200 20 ml. A 300 ml. portion of ethanol is added and the produce is allowed to crystallize, giving 25.1 g. of 4-hydroxy-6--nitro-2-naphthoic acid ethyl ester acetate as pale yellow crystals. A mixture of 26.3 g. of 4-hydroxy-6-nitro-2-naph-25 thoic acid ethyl ester acetate, 250 ml. of tetrahydrofuran and 2.5 g. of 10% palladium on carbon is hydrogenated on a Parr shaker at 40-20 psi over 45 minutes. The mixture is filtered through diatomaceous earth and evaporated _in vacuo to an oil. This oil is crystallized from 200 ml. of ether, 30 giving 21.8 g. of 6-amino-4-hydroxy-2-naphthoic acid ethyl ester acetate as beige crystals. To a cooled (ice bath) solution of 9.02 g. of 6--amino-4-hydroxy-2-naphthoic acid ethyl ester acetate in 50 ml. of dry pyridine is added 6.74 g. of m-nitrobenzoyl chloride. After 5 minutes the ice bath is removed and PATEWTo^Oi stirring is continued at room temperature for 30 minutes. The solution is poured into 500 ml. of water and stirred 2 7 APR 1983 fvECBVb •| Q 7 .< A i / O i * - 12 - until the precipitate solidifies. The solid is collected by filtration, washed with water, dried and crystallized from 250 ml. of acetonitrile at 5°C., giving 13.0 g. of 4-hydroxy-6-m-nitrobenzamido-2-naphthoic acid ethyl ester 5 acetate as beige crystals. A mixture of 13.0 g. of 4-hydroxy-6-m-nitrobenza-mido-2-naphthoic acid ethyl ester acetate, 125 ml. of tetrahydrofuran and 1.25 g. of 107o palladium on carbon is hydrogenated in a Parr shaker at 45-37 psi for one hour. 10 The mixture is filtered through diatomaceous earth and the filtrate is evaporated _in vacuo to a pale yellow glass. This glass is crystallized by trituration with ether and the solid is recrystallized from 100 ml. of acetonitrile at 5°C. , giving 10.35 g. of the desired product as colorless 15 crystals, m.p. 185-187°C. Example 2 6 ,6 ' - [ Ureylenebis Tin-phenylenecarbonyl imino) ]bis- [4-hydroxy-2-naphthoic acid]diethyl ester diacetate To a solution of 10.23 g. of 6-(m-aminobenzamido)-20 -4-hydroxy-2-naphthoic acid ethyl ester acetate in 60 ml. of dry pyridine is added a solution of 1.3 g. of phosgene in 5 ml. of dry ethylene glycol dimethyl ether, dropwise, with stirring and cooling, during 2-3 minutes. Stirring is continued at room temperature for 2 hours, then the 25 solution is poured into 800 ml. of water. The gummy precipitate is triturated with water, giving a red solid. This solid is stirred and refluxed in 300 ml. of ethanol, cooled, filtered and the solid is washed with ethanol, then ether. This solid is dissolved in 120 ml. of hot dimethyl-30 formamide, treated with charcoal and filtered through diatomaceous earth. The filtrate is warmed to 80°C. and diluted slowly, with stirring with 60 ml. of water. The mixture is cooled to room temperature and the solid is collected by filtration, washed with 67% aqueous dimethylformamide, ethanol, then ether and dried overnight at 110°C., giving PATENTnpflcy8.45 g. of the desired product as a pale tan powder, m.p. 27APR1983 285"287°c* REi.£'iV*i: i) \ 9 6 7 6 4 - 13 - Example 3 6,6'-[Ureylenebis(m-phenylenecarbony1imino)]-bis[4-hydroxy-2-naphthoic acicQ To a cooled (water bath)( solution of 4.06 g. of 5 6,6' - [ureylenebis (m-phenylenecarbonylimino) ]bis [4-hydroxy-. .J . — -2-naphthoic acidjdiethyl ester diacetate in 60 ml. of di-methylsulfoxide is added 60 ml. of 2N sodium hydroxide, por-tionwise, with stirring, in a nitrogen atmosphere. The mixture is stirred under nitrogen at room temperature for 2 10 hours, then poured into 300 ml. of water and filtered. The filtrate is acidified to pH 2 with the addition of 10 ml. of concentrated hydrochloric acid and 50 g. of sodium acetate trihydrate are added. The gel is filtered and washed with water, then further washed with water in a centrifuge and 15 dried by co-evaporation with 750 ml. of n-propanol, giving a red-brown powder. This powder is dissolved in 25 ml. of hot dimethylformamide, diluted slowly with 15 ml. of water and cooled in a refrigerator. The precipitate is collected by filtration, washed successively with 8 ml. of 50% aqueous 20 dimethylformamide, ethanol:ether (1:1) and finally ether, then dried overnight at 110°C., giving 1.6 g. of the desired product as a pale tan powder, m.p. 297-300°C. (dec.). Example 4 6,6'-[Ureylenebis(m-phenylenecarbonylimino)]-25 bis[4-hydroxy-2-naphthoic acid] diethyl ester To a cooled (water bath) solution of 2.0 g. of 6 , 6 ' -[ureylenebis(m-phenylenecarbonylimino)]bis[4-hydroxy-2--naphthoic acid] diethyl ester diacetate in 60 ml. of dimethylformamide is added 40 ml. of 0.25N sodium hydroxide, drop-30 wise with stirring over 10 minutes. The solution is stirred for an additional 10 minutes, 80 ml. of pyridine is added and the solution is poured with cooling into a mixture of 800 ml. of water and 85 ml. of concentrated hydrochloric acid. The solid is collected by filtration, washed with water and dried at room temperature. This solid is dissolved NX PATENT OFFICE 2 7 APR 1983 RECEIVED c tj t in hot 2-methoxyethanol ac a concentration of 4% (w/v). This solution is then distilled with 1/2 its volume of water and then cooled to room temperature. The precipitate is collected by filtration and washed with 50% aqueous 2-methoxyethanol, ethanol, then ether. This solid is dis-5 solved in a mixture of 2-methoxyethanol .-dimethyl formamide (6:5:1) giving an approximate 4% (w/v) solution, diluted with water and filtered. The solid is washed as described above, then with acetone and dried at 110°C., overnight giving 417 mg. of the desired product as a tan powder, m.p. 10 265-280°C. (dec.). Example 5 Preparation of Compressed Tablet Ingredient mg./Tablet Active Compound 0.5-500 15 Dibasic Calcium Phosphate N.F qs Starch U. S. P 40 Modified Starch 10 Magnesium Stearate U.S.P 1-5 Example 6 20 Preparation of Compressed Tablet - Sustained Action Ingredient mg./Tablet Active Compound as Aluminum 0.5-500 (as acid Lake*, Micronized Dibasic Calcium Phosphate N.F qs 25 Alginic Acid 20 Starch U.S.P 35 Magnesium Stearate U.S.P 1-10 ^Collagenase inhibitor plus aluminum sulfate yields aluminum collagenase inhibitor. Collagenase inhibitor 3q content in aluminum lake ranges from 5-30%. Example 7 Erfiflflrafinn of Hard Shell Capsule Ingredient mg./Capsule Active Compound 0.5-500 Lactose, Spray Dried qs Magnesium Stearate 1-10 10 15 20 25 30 1 /? 4 IT - Example 8 Preparation of Oral Liquid (Syrup) Ingred ient Active Compound Liquid Sugar Methyl Paraben U.S.P Propyl Paraben U.S.P Flavoring Agent Purified Water qs ad Example 9 7o W/V 0.05-5 75.0 0.18 0.02 qs 100.0 Preparation of Oral Liquid (Elixir) Ingredient % W/V Active Compound 0.05-5 Alcohol U.S.P 12.5 Glycerin U.S.P 45.0 Syrup U.S.P 20.0 Flavoring Agent qs Purified Water qs ad 100.0 Example 10 Preparation of Oral Suspension (Syrup) Ingredient % W/V Active Compound as Aluminum 0.05-5 Lake, Micronized (acid equivalent) Polysorbate 80 U.S.P Magnesium Aluminum Silicate, Colloidal 0.1 0.3 Flavoring Agent qs Methyl Paraben U.S.P 0-18 Propyl Paraben U.S.P 0.02 Liquid Sugar 75.0 Purified Water qs ad 100.0 Example 11 Preparation of Injectable Solution Ingredient % W/V Active Compound 0.05-5 Benzyl Alcohol N.F 0.9 Water for Injection 100.0 lb - JrT- - Example 12 Preparation of Injectable Oil Ingred ient % W/V Active Compound 0.05-5 Benzyl Alcohol 1.5 Sesame Oil qs ad 100.0 Example 13 Preparation of Intra-Articular Product Ingredient Amount Active Compound 2-20 mg . NaCl (physiological saline) 0.9% Benzyl Alcohol 0.9% Sodium Carboxymethylcellulose 1.5% pH adjusted to 5.0-7.5 Water for Injection qs ad 100% Example 14 Preparation of Injectable Depo Suspension Ingred ient % W/V Active Compound 0.05-5 (acid equivalent) Polysorbate 80 U'_S».P 0.2 Polyethylene Glycol 400 U.S.P 3.0 Sodium chloride U.S.P 0.8 Benzyl Alcohol N.F 0.9 HC1 to pH 6-8 qs Water for Injection qs ad 100.0 Example 15 Preparation of Dental Paste Ingredient % W/V Active Compound 0.05-5 Zinc Oxide 15 Polyethylene Glycol 4000 U.S.P. .. 50 Distilled Water qs 100 Example 16 Preparation of Dental Ointment Ingredient % W/V Active Compound 0.05-5 -J- Petrolatum, White U.S.P. qs 100 Example 17 Preparation of Dental Cream Ingred ient % W/V Active Compound 0.05-5 Mineral Oil 50 Beeswax 15 Sorbitan Monostearate 2 Polyoxyethylene 20 Sorbitan Monostearate 3 Methyl Paraben U.S.P 0.18 Propyl Paraben U.S.P 0.02 Distilled Water qs 100 Example 18 Preparation of Topical Cream Ingredient % W/V Active Compound 0.05-5 Sodium Lauyl Sulfate 1 Propylene Glycol 12 Stearyl Alcohol 25 Petrolatum, White U.S.P 25 Methyl Paraben U.S.P 0.18 Propyl Paraben U.S.P 0.02 Purified Water qs 100 Example 19 Preparation of Topical Ointment Ingredient % W/W Active Compound 0.05-5 Cholesterol 3 Stearyl Alcohol 3 White Wax 8 Petrolatum, White U.S.P. qs 100 Example 20 Preparation of Spray Lotion (Non-Aerosol) Ingredient % W/W Active Compund 0.05- Isopropyl Myristate 20 Alcohol (Denatured) qs 100 «£ i if - kS - Example 21 Preparation of Buccal Tablet Ingred ient g./Tablet Active Ingredient 0.00325 5 6 x Sugar 0.29060 Acacia 0.01453 Soluble Starch 0.01453 F. D. & C. Yellow No. 6 Dye 0.00049 Magnesium Stearate 0.00160 10 0.325O0 The final tablet will weigh about 325 mg. and may be compressed into buccal tablets in flat faced or any other tooling shape convenient for buccal administration. Example 22 15 Preparation of Lozenge Ingredient g./Lozenge Active Ingredient 0.0140 Kompact® Sugar (Sucrest Co.) 0.7138 6 x Sugar 0.4802 2o Sorbitol (U.S.P. Crystalline) 0.1038 Flavor 0.0840 Magnesium Stearate 0.0021 Dye qs Stearic Acid 0.0021 1.4000 25 The ingredients are compressed into 5/8" flat based lozenge tooling. Other shapes may also be utilized. Example 23 Preparation of Gelled Vehicles Ingredient % W/W Active Compound 9-11 Sodium Chloride 0.9-1.2 Buffer and Flavor qs Purified Water qs ad 100 Ingredient % W/W Active Compound ~ 0.005-9 Sodium Alginate 0.5-2 Buffer and Flavor qs...., 30 1 r V w 4 'i X- Purified Water qs ad 100 Ingredient % W/W Active Compound 0.005-9 Hydroxypropy1 Cellulose 0.5-2 5 Buffer and Flavor qs Purified Water qs ad 100 Ingred ient % W/W Active Compund 0.005-9 Guar Gum 0.5-2 10 Buffer and Flavor qs Purified Water qs ad 100 Example 24 Preparation of Oral MoUth Rinse Ingredient % W/V 15 Active Compound 0.05-20 Alcohol U.S.P 0.20 Sorbitol 1-30 Buffer and Flavor qs Polysorbate 80 0.1-3 20 Cetyl Pyridinium Chloride 0.025-0.20 Purified Water qs ad 100 Example 25 Preparation of Tooth Paste Ingredient % W/W 25 Active Compound 0.05-15 Glycerin 5-15 Sorbitol 5-15 Sodium Carboxymethylcellulose 0.5-2 Magnesium Aluminum Silicate 0.1-1 30 Carrageenin 0.25-2 Preservative qs Sodium Lauryl Sulfate 0.1-3 Calcium Carbonate 1 25-45 Flavor qs Purified Water qs ad 100 Example 26 Preparation of Dental Paste r; s- <10 - - Ingredient % W/W Active Compound 0.05-20 Carboxymethylcellulose 5-20 Pectin 5-20 5 Plastibase® 20-70 Gelatin 5-20 Example 27 Preparation of Dental Ointment Ingredient % W/W 10 Active Compound 0.05-20 Polyethylene Glycol 4000 50-80 Polyethylene Glycol 400 10-40 Example 28 Preparation of Dental Powder for Brushing or 15 for Use in Water Spray (e.g. Water Pik®) Ingredient % W/W Active Compound 0.05-10 Flavor qs Wetting Agents....qs 20 Dextrin qs ad 100 Example 29 Preparation of Stick for Application to Gums Ingredient % W/W Active Compound 0..05-10 25 Glycerin 5-10 Propylene Glycol 40-80 Sodium Stearate 6-10 Flavor qs Water 0-10 30 Example 30 Preparation of Periodontal Packing Paste Paste Part A Ingredient % W/W Active Compound 0.05-20 Caprylic Acid 9.0 Laurie Acid 27.0 Ethylcellulose (100 cps.) 2.0 4 ***•/? '-•■7 /" A 1 ^ j . o 4 aA - 2? - Polypale Resin- 39.0 Gum Elemi 4.0 Brominol** 4.0 Mica (Powdered) 7.5 5 Chlorothymol 1.0 Zinc Acetate 2.0 Bay Oil (Essential Oil) 1.0 Ethanol 1.5 Paste Part B 10 Magnesium Oxide 43.0 Zinc Oxide 21.0 Calcium Hydroxide 3.5 Copper Oxide 2.0 Mineral Oil, Heavy 26.0 15 Rosin Oil 3.0 Chlorothymol 1.4 Cumarin (Flavor) 0.1 ""Partially polymerized rosin (i.e. modified rosin) **Brominated olive oil 20 When equal parts of A and B are mixed together at 25°C. a hard mass is formed in about 3 minutes. Example 31 Preparation of Periodontal Packing Paste Part A (Powder) 25 Ingredient % W/W Active Compound 0.05-20 Canada Balsam, Neutral 8.5 Rosin NF 8.5 Calcium Hydroxide 34.4 30 Zinc Oxide U.S.P 46.6 Part B (Liquid Hardener) Eugenol 85.0 Turpentine Oil, Rectified 15.0 A mixture of three drops of Part B added to 130 mg. of Part A produces a hard mass in about 2-3 minutes at 30°C. The compounds of this invention may be administered internally to a warm-blooded animal to inhibit connective 19676 - 22 - - - J •\ " " "* I tissue-destruction or collagenase, such inhibition'being -useful in the amelioration or prevention of those reactions causing connective tissue damage. A range of doses may be employed depending on the mode of administration, the condition being treated and the particular compound being used. For example, for intravenous or subcutaneous use from about 5 to about 50 mg./kg./day, or every six hours for more rapidly excreted salts, may be used. For intra-articular use for large joints such as the knee, from about 2 to about 20 mg./joint per week may be used, with proportionally smaller doses for smaller joints. The dosage range is to be adjusted to provide optimum therapeutic response in the warm-blooded .animal being.treated. ! In general, the amount of compound administerred can vary over a wide range to provide from about 1.5 mg./kg. to about 100 mg./kg. of body weight of animal per day. The usual daily dosage for a 70 kg. subject may vary from about 100 mg. to about 3.5 g. Unit doses can contain from about 0.5 mg. to about 500 mg. While in general the sodium salts-of the acids of the invention are suitable for parenteral use, other salts may also be prepared, such as those of primary amines, e.g., ethylamine; secondary amines, e.g., diethylamine or diethanolamine; tertiary amines, e.g., pyridine or triethylamine or 2-dimethylaminomethyldibenzofuran; aliphatic diamines, e.g., decamethylenediamine; and aromatic diamines, can be prepared. Some of these are soluble in water, others are soluble in saline solution, and still others are insoluble and can be used for purposes of preparing suspensions for injection. Furthermore, as well as the sodium salt, those of the alkali metals, such as potassium and lithium; of ammonia; and of the alkaline earth metals, such as calcium or magnesium, may be employed. It will-be apparent, therefore, that these salts embrace, in general, derivatives of salt-forming cations. In therapeutic use the compounds of this inven- -tion may be administered in the form of conventional 1 C /• ' r~7 *•' O , ' fb - 2A - pharmaceutical compositions. Such compositions may be formulated so as to be suitable for oral or parenteral administration. The active ingredient may be combined in admixture with a pharmaceutically acceptable carrier, which carrier may take a wide variety of forms depending on the form of preparation desired for administration, i.e., oral or parenteral. The compounds can be used in compositions such as tablets. Here, the principal active ingredient is mixed with conventional tabletting ingredients such as corn starch, lactose, sucrose, sorbitol, talc, stearic acid, magnesium stearate, dicalcium phosphate, gums, or similar materials as non-toxic pharmaceutically acceptable diluents or carriers. The tablets or pills of the novel compositions can be laminated or otherwise compounded to provide a dosage form affording the advantage of prolonged or delayed action or predetermined successive action of the enclosed medication. For example, the tablet or pill can comprise an inner dosage and an outer dosage component, the latter being in the form of an envelope over the former. The two components can be separated by an enteric layer which serves to resist disintegration in the stomach and permits the inner component to pass intact into the duodenum or to be delayed in release. A variety of materials can be used for such enteric layers or coatings, such materials including a number of polymeric acids or mixtures of polymeric acids with such materials as shellac, shellac and cetyl alcohol, cellulose acetate and the like. A particularly advantageous enteric coating comprises a styrene maleic acid copolymer together with known materials contributing to the enteric properties of the coating. The tablet or pill may be colored through the use of an appropriate non-toxic dye, so as to provide a pleasing appearance . The liquid forms in which the novel compositions of the present invetion may be incorporated for administration include suitable flavored emulsions with edible oils, such as, cottonseed oil, sesame oil, coconut oil, a? - 25 - peanut oil, and the like, as well as elixirs and similar pharmaceutical vehicles. Sterile suspensions or solutions can be prepared for parenteral use. Isotonic preparations containing suitable preservatives are also desirable for 5 injection use. The compounds of the present invention may also be administered topically in the form of ointments,- creams, lotions and the like, suitable for the treatment of connective tissue dependent dermatological disorders. 10 Moreover, the compounds of the present invention may be administered in the form of dental pastes, ointments, buccal tablets and other compositions suitable for application periodontally for the treatment of periodontitis and related diseases of the oral cavity. 15 The term dosage form as described herein refers to physically discrete units suitable as unitary dosage for warm-blooded animal subjects, each unit containing a predetermined quantity of active component calculated to produce the desired therapeutic effect in association with 20 the required pharmaceutical diluent, carrier or vehicle. The specification for the novel dosage forms of this invention are indicated by characteristics of the active component and the particular therapeutic effect to be achieved or the limitations inherent in the art of compounding 25 such an active component for therapeutic use in warm-blood-ed animals as disclosed in this specification. Examples of suitable oral dosage forms in accord with this invention are tablets, capsules, pills, powder packets, granules, wafers, cachets, teaspoonfuls, dropperfuls, ampules, 30 vials, segregated multiples of any of the foregoing and other forms as herein described. The inhibiting activity of representative compounds of the invention on the destruction of connective tissue has been demonstrated by one or more of the following identified tests: (i)Collagen Assay, Test Code 006 -This test measures the ability of human skin fibroblast collagenase to degrade radiolabeled native collagen fi- 196764 - 25- - brils. An active inhibitor inhibits the degradation of the collagen fibril; (ii) Crevicular fluid Assay - In an analogous test, collagenase present in the crevicular fluid of inflamed gingival tissue was used to measure its ability to degrade radiolabeled native collagen fibrils. An active inhibitor would inhibit the degradation of the collagen fibril; (iii) Leukocyte Neutral Proteases Inhibitor Assay - This test measures the ability of neutral proteases derived from human leukocytes to degrade radiolabeled proteoglycans entrapped in polyacrylamide beads. An active inhibitor inhibits the degradation of proteoglycans. (i) Collagenase Assay - Test Code 006 Collagenase assays were performed by a modification of the method of Harper, e_t al_. , Biochem. J., 10_, 3035 (1971). In a typical assay (total volume of 0.45 ml.), 100 ul. of the activated enzyme was added to the C-labeled collagen fibrils (250 ul.) followed by 100 ul. of 50 mM cacocylate, pH 7.4, containing 5 mM calcium chloride. After incubation at 37°C. for 16 hours, the tubes were centri-fuged in a Beckman microfuge for five minutes at full speed. An aliquot (200 ul.) of the supernatant, representing collagenase digestion products of the fibril, was assayed for radioactivity. The effect of the test compound on collagen degradation by collagenase was examined as follows: The test compound (in distilled water) was added at a test concentration of 30 ug./ml. to the assay tubes containing active collagenase (total volume 450 ul.) and after 16 hours the amount of radioactivity in the supernatant was determined. Appropriate blanks and trypsin controls were run in parallel. Table I shows that representative compounds of the invention possess collagenase inhibitory activity. The activities are expressed as % inhibition (lowering) of collagenase activity, i.e. based on the 0% value for the enzyme con-rol. </div>

Claims

<div class="application article clearfix printTableText" id="claims"> - jtr - Table I Biological Activities Compound % Inhibition of Collagenase 6 ,6 ' -[Ureylenebis(m-phenylenecarbonyl imino)]bis[4-hydroxy-2-naphthoic acid] diethyl ester d iacetate 7 6,61-[ Ureylenebis(m-phenylenecarbonyl imino)]bis[4-Hydroxy-2-naphthoic acid] 92 6 ,6 1 -[Ureylenebis(m-phenylenecarbonyl imino)]bis[4-Hydroxy-2-naphthoic acid] diethyl ester 57 27 •: o : 7 •: ,i, WHAT WE CLAIM IS:

1. A compound of the formula: HCO CO wherein A is selected from the group consisting of hydrogen, lower (C1-C5) alkyl and a pharmaceutically acceptable salt cation; B is selected from the group consisting of hydrogen, lower (C1-C5) alkanoyl and a pharmaceutically acceptable salt cation; and R is selected from the group consisting of hydrogen and lower (C1-C3)alkyl.

2. A compound according to Claim 1 of. the formula: NHCO NHCO wherein A is selected from the group consisting of hydrogen, lower (C1-C5)alkyl and a pharmaceutically acceptable salt cation; B is selected from the group consisting ofhydrogen, lower (C1-C5)alkanoyl and a pharmaceutically acceptable acceptable salt cation; and R is selected from the group consisting of hydrogen and lower (C1-C3)alkyl.

3. A compound according to Claim 1 of the formula: "7 O i - 28 - wherein A is selected from the group consisting of hydrogen, lower (C-j^-Cg) alkyl and a pharmaceutically acceptable salt cation; B is selected from the group consisting of hydrogen, lower (C^-Cg)alkanoyl and a pharmaceutically acceptable salt cation; and R is selected from the group consisting of hydrogen and lower (C^-C^)alkyl.

4. The compound according to Claim 1, 6,6'-[ureylenebis (m-phenylenecarbonylimino)]bis[4-hydroxy-2-naphthoic acid]diethyl ester diacetate, 6,61 -[ureylenebis(m-phenylenecarbonylimino) ] bis [4-hydroxy-2-naphthoic acid], or 6,6'-[ureylenebis(m-phenylenecarbonylimino)]bis[4-hydroxy--2-naphthoic acid] diethyl ester.

5. A method for the preparation of a compound of the formula: - 29 - I ° ^ < .1 I / u / O '-!• wherein A is selected from the group consisting of hydrogen, lower (C^-Cg)alkyl and a pharmaceutically acceptable salt cation; B is selected from the group consisting of hydrogen, lower (C^-Cg)alkanoyl and a pharmaceutically acceptable salt cation; and R is selected from the group consisting of hydrogen and lower (C^-C^)alkyl; which comprises reacting a sub-stituted-aminonaphthoic acid of the formula: wherein A and B are as previously defined; in a basic solution with an excess of a substituted nitrobenzoylchloride of the formula: wherein R is as previously defined; to give a substituted nitrobenzamido-substituted naphthoic acid of the formula: AO* COCl NO 2 NHCO BO AOOC then hydrogenating said substituted nitrobenzamido-substit-uted naphthoic acid to give the corresponding amine derivative of the formula: - 30 - ' o "7 '■ 1 s O / O ', NH2 NHCO R and phosgenating said amine derivative.

6. A composition of matter in dosage unit form which comprises an effective connective tissue destruction inhibiting amount from substantially 0.5 mg to substantially 500 mg of a compound of the formula: NHCO wherein A is selected from the group consisting of hydrogen, lower (C-^-Cg) alkyl and a pharmaceutically acceptable salt cation; B is selected from the group consisting of hydrogen lower (C^-C^) alkanoyl and a pharmaceutically acceptable salt cation; and R is selected from the group consisting of hydrogen and lower (C-^-C^) alkyl; in association with a pharmaceutically acceptable carrier. p M ? PATEHT 1 -1 NOV 19S3 - 31 - I V O / O -

7. The composition of matter according to Claim 6, wherein the compound is 6,61-[ureylenebis(m-phenylenecarbonylimino) ] bis [ 4-hydroxy-2-naphthoic acid]diethyl ester diacetate; 6,6'-[ureylenebis(m-phenylenecarbonylimino)]bis[4-hydroxy-2-naphthoic acid]; or 6,6'-[ureylenebis(m-phenylenecarbonylimino) ]bis[4-hydroxy-2-naphthoic acid]diethyl ester. DATED THIS /f/t DAY OF 1^? A . J . P A R K & SOW PER v// ^ ' •>£-*-<'* > AGENTS FOR THE APPLICANTS N.Z. PAT' ' </div>