COMPOUND HAVING ANTIBRONCHOPNEUMOPATHIC ACTIVITY, PROCESS FOR ITS PREPARATION AND PHARMACEUTICAL COMPOSITIONS CONTAINING IT

Abstract: 4[[(2-(2-Thenoyl)amino-3,5-dibromophenyl)-methyl]-­ amino]cyclohexanol of formula (I): and pharmaceutically acceptable acid addition salt thereof have been found to have valuable therapeutic properties. Compound (I) may be prepared by reacting 4-[[(1-amino-­ 3,5-dibromophenyl)-methyl]amino]cyclohexanol with thenoyl chloride.

Description: Compound having antibronchopneumopathic activity, process for its preparation and pharmaceutical compositions containing it The present invention relates to 4-/ /(2-(2 thenoyl)-amino-3,5-dibromophenyl)-methyl/amino/cy- clohexanol of formula I EMI1.1 to a process for preparing it and to pharmaceutical compositions containing it as the active principle. The invention relates also to pharmaceutically acceptable acid addition salts of compound I. The hydrochloric salt of compound I, which will be hereinafter called PU 163, for sake of shortness, shows valuable pharmacological and pharmaco-kynetic properties and it is thus effective for the treatment both of acute and chronic respiratory diseases in humans, especially in case of qualitative or quan titative impairments in the mucous secretion, such as in bronchitis, bronchopneumonitis, bronchial asth ma, pulmonary emphysema, tracheobronchitis, and the like. The compound I proved to have better effectiveness and pharmacokynetis behaviour in comparison with known mucolytic drugs, namely Ambroxol. Compound I may be prepared by reacting 4-/ /(2amino-3,5-dibromophenyl)-methyl/amino/cyclohexanol (Ambroxol), a well known compound (vide U.S. Patent No. 3,536,713) currently used in therapy, with 2thenoyl chloride in inert solvents, in the presence of acid-binding agents. The nature of the solvent is not particularly critical, provided that it has no adverse effect upon the reaction; suitable solvents include benzene, toluene and pyridine. Alkali carbonates or bicarbonates, tertiary amines etc. may be used as acid-binding agents. The reaction temperature is not critical, and it may range from +IOOC and the reflux temperature of the solvent. The acid addition salts may be obtained by conventional salification methods, for instance the hydrochloric salt may be added by bubbling gaseous hydrochloric acid in an ethanol solution of compound I. The following example further illustrates the process according to the invention, without limiting it. EXAMPLE To a solution of Ambroxol in the free form (39 g; 0.1 mole) in toluene (350 ml) triethylamine (14 ml; 0.1 mole) was added, then 2-thenoyl chloride (14.6 ml; 0.1 mole) was added dropwise at room temperature. The reaction mixture was left to stand overnight, then the toluene solution was washed with wa ter and the solvent was evaporated off. The resulting residue (45 g) was dissolved in ethanol (250 ml) and gaseous hydrochloric acid was added to acid pH. From the solution the hydrochloric salt precipi tated, which was filtered and washed with ethanol, yielding 43 g of the desired compound; m.p. = 282-2860C. Elemental analysis (for C18H20Br2N2O2S.HCl, PM = 524.71) C H N S Calc. % 41.20 3.84 5.34 6.11 Found % 40.97 3.91 5.27 6.10 The NMR data agree with the structure of the com pound. NMR (6) : 1.1-2 (8H, m); 2.8-3.6 (3H, m); 3.8-4.2 (1H, m); 4.3-4.7 (1H, m); 7.2 (1H, t); 7.8-8.1 (3H, m); 8.3 (1H, d); 9.6 (2H, m); 10.3-10.7 (1H, m). The toxico-pharmacological characteristics of compound I (PU 163) are documented by the following tests. Ambroxol, active principle of several pharmaceu tical specialties, which is 4-/ /(2-amino-3,5-dibromophenyl)-methyl/amino/cyclohexanol (Merk Index, tenth edition, 383), was used as reference drug. Acute toxicity The acute toxicity of PU 163 has been determined in the mouse and in the rat by the oral route accor ding to the method of Litchfield and Wilcoxon (J. Pharm Exp. Therap. 1949, 96, 99). The results are reported in the following Table 1. TABLE 1 - Acute toxicity EMI4.1 <tb> Species <SEP> Administration <SEP> route <SEP> LD <SEP> - <SEP> mg/kg <tb> <SEP> Ambroxol <SEP> PU <SEP> 163 <tb> irouse <SEP> os <SEP> > <SEP> 2,500 <SEP> ?2,500 <SEP> <tb> rat <SEP> os <SEP> > 5,000 <SEP> > 5,000 <tb> Pharmacology Action on the ciliary motility in the dog's trachea The mucolytic activity of many products is due in that they induce an increase of motility in the trachea ciliate cells. The method hereinafter described allows to assess "in vitro" if a compound increases, or not, the motility of said cells. The method is based on the measurement of the transit speed of a charcoal bolus in a section of trachea. The method used is that described by J.E. Lightowler and N.H. Lightowler (Arch. Int. Pharmacodyn. 189, 53-58, 1971) and uses an apparatus similar to that described by T. Dalhamn (Acta Physio Scand. Suppl. 123, 1956). PU 163 has been used in comparison with Ambroxol, at the same concentration of 10 4 g/ml and the results deriving from the test are reported in Table 2, from which'it can be shown that PU 163 exhibits an action on the ciliary motility to an higher extent than that of Ambroxol, but of the same order of significance. TABLE 2 - Mucolytic activity - Ciliary motility on the dog's trachea. EMI6.1 TREATMENT <SEP> CONCENTRATION <SEP> N <SEP> SPEED <SEP> OF <SEP> CHARCOAL <SEP> PARTICLES <tb> <SEP> g/ml <SEP> ANIMALS <tb> <SEP> Basal <SEP> 5 <SEP> Minutes <SEP> 10 <SEP> Minutes <tb> <SEP> mm/min. <SEP> mm/min. <SEP> % <SEP> # <SEP> mm/min. <SEP> % <SEP> # <SEP> mm/min. <SEP> % <SEP> # <tb> <SEP> x <SEP> # <SEP> ES <SEP> x <SEP> # <SEP> ES <SEP> vs. <SEP> Bas. <SEP> x <SEP> # <SEP> ES <SEP> vs. <SEP> Bas. <SEP> x <SEP> # <SEP> ES <SEP> vs. <SEP> Bas. <tb> <SEP> x <SEP> # <SEP> ES <SEP> x <SEP> # <SEP> ES <SEP> x <SEP> # <SEP> ES <tb> CONTROLS <SEP> ¯¯ <SEP> 4 <SEP> 5.2 <SEP> 6.3 <SEP> 21.1 <SEP> 6.02 <SEP> 19.2 <SEP> 5.8 <SEP> 11.5 <tb> <SEP> #1.03 <SEP> #1.36 <SEP> #3.38 <SEP> #1.07 <SEP> # <SEP> 6.25 <SEP> #1.04 <SEP> # <SEP> 3.82 <tb> AMBROXOL <SEP> 10-4 <SEP> 4 <SEP> 6.4 <SEP> 8.2 <SEP> 28.12 <SEP> 9.5 <SEP> 57.6* <SEP> 10.2 <SEP> 59.37** <tb> <SEP> #1.30 <SEP> #2.12 <SEP> #3.54 <SEP> #1.62 <SEP> #14.60 <SEP> #2.31 <SEP> # <SEP> 8.12 <tb> PU <SEP> 163 <SEP> 10-4 <SEP> 4 <SEP> 7.00 <SEP> 8.7 <SEP> 24.28 <SEP> 10.2 <SEP> 45.7* <SEP> 11.5 <SEP> 64.28** <tb> <SEP> #1.40 <SEP> #2.82 <SEP> #7.80 <SEP> #2.02 <SEP> # <SEP> 8.40 <SEP> #2.38 <SEP> # <SEP> 9.12 <tb> student's "t" test versus buffer solution* p < 0.05 ** p < 0.01 *** p < 0.001 Bronchosecretogogue activity The bronchosecretogogue activity of PU 163 and, as a comparison, of Ambroxol has been determined in the rat using the method described by Mawatari ("Experimental Studies on the Expectorant Action of Several Drugs", Kagoshima Daigaku Igaku Zasshi, 27, 561, 1976). The compounds under exam have been administered by the oral route at equiponderal doses of 50 mg 30 minutes after the subcutaneous administration of sodium fluorescein. After the animals'sacrifice, the evaluation of the amound of sodium fluorescein present at the bronchial level has been carried out. The results obtained, reported in the Table 3, are expressed as percent increase of the bronchial elimination of sodium fluorescein of the groups treated with Ambroxol and PU 163 in comparison with a control group. As it can be seen, PU 163 showed an high bronchosecretogogue activity, in slightly higher extent than that of Ambroxol in the used experimental conditions. TABLE 3 - Bronchosecretogogue activity in the rat (Mawatari test) EMI8.1 <tb> <SEP> TRATMENT <SEP> DOSE <SEP> N <SEP> OF <SEP> ANIMALS <SEP> SODIUM <SEP> FLUORESCEIN <tb> <SEP> mg/kg <SEP> ANI- <SEP> WEIGHT/ <SEP> g/ml <SEP> % <SEP> vs. <tb> <SEP> i.p. <SEP> MALS <SEP> grams <SEP> control <tb> CONTROLS <SEP> - <SEP> 10 <SEP> 96.0 <SEP> 0.28 <SEP> <tb> <SEP> #5.56 <SEP> <SEP> #0.02 <tb> AMBREXOL <SEP> 50 <SEP> 10 <SEP> 98.0 <SEP> 0.37* <SEP> 32.1 <tb> <SEP> +7.41 <SEP> +0.05 <SEP> <tb> PU <SEP> 163 <SEP> 50 <SEP> 10 <SEP> 102.0 <SEP> 0.39* <SEP> 39.3 <tb> <SEP> #2.91 <SEP> <SEP> #0.06 <tb> Student's "t" test *p < 0.05 Activity on the alveolar surfactant This test allow to verify if a compound can modi fy the surfaces of lung alveoli, as there are pathological conditions in which a superficial firm is produced in the alveoli which, as time goes on, can produce foam in the bronchi, making the respiration difficult. In order to assess this kind of activity, the method described by Curty P.C. (Arzneim. Forsch. Drug. Res., 24, 847, 1974) is used. According to said method the volume and the pressure after insufflation in the rat's lung, are measured. PU 163 and Ambroxol as reference drug have been administered at equiponderant doses of 50 mg/kg in the rat by oral route, three hours before the test. After sacrifice, and after the trachea incan nulation, the respiratory apparatus has been extracted from the thoracic cavity and the static cur ves of volume-pulmonary pressure. From the test results, as it is shown in the fol lowing Table 4, it is found that both the compounds under exam decrease the pulmonary pressure induced by air insufflation in marked way in comparison with the control animals. At the same time, however, the recovery of the basal pressure in the insufflation phase is shorter for PU 163 both in comparison with the controls and with Ambroxol and in the de-insufflation phase the recovery of PU 163 is longer both in comparison with the controls and in comparison with Ambroxol. Therefore, in the used experimental conditions, PU 163 seems to be endowed with an higher activity on the test of the alveolar surfactant. TABLE 4 - Activity of the alveolar surfactant in the rat - Static insufflation and de-insufflation rates. EMI10.1 Treatment <SEP> Dose <SEP> N <SEP> of <SEP> x <SEP> Internal <SEP> pulmonary <SEP> pressure <SEP> (cm. <SEP> H2O) <tb> <SEP> mg/kg <SEP> animals <SEP> Weight <tb> <SEP> p.o. <SEP> g <SEP> 0.5 <SEP> 1 <SEP> 1.5 <SEP> 2 <SEP> 2.5 <SEP> 3 <SEP> 3.5 <SEP> 3 <SEP> 2.5 <SEP> 2 <SEP> 1.5 <SEP> 1 <SEP> 0.5 <SEP> cm <tb> <SEP> 266.0 <tb> Controls <SEP> - <SEP> 5 <SEP> #5.10 <SEP> 3 <SEP> 5 <SEP> 6.65 <SEP> 8.3 <SEP> 10 <SEP> 10.7 <SEP> 11.6 <SEP> 7.8 <SEP> 5.3 <SEP> 3.2 <SEP> 1.4 <SEP> 0.25 <SEP> <SEP> 245.0 <tb> Ambroxol <SEP> 50 <SEP> 5 <SEP> #7.20 <SEP> 2.45 <SEP> 4.1 <SEP> 5.6 <SEP> 6.9 <SEP> 7.9 <SEP> 8.7 <SEP> 9.3 <SEP> 6.1 <SEP> 4.6 <SEP> 3.3 <SEP> 2.3 <SEP> 1.1 <SEP> 0.25 <tb> <SEP> 238.0 <tb> PU <SEP> 163 <SEP> 50 <SEP> 5 <SEP> #5.00 <SEP> 2.2 <SEP> 2.8 <SEP> 5.35 <SEP> 6.5 <SEP> 7.6 <SEP> 8.45 <SEP> 9 <SEP> 6.75 <SEP> 5.2 <SEP> 3.9 <SEP> 2.5 <SEP> 1.4 <SEP> 0.35 <tb> Action on the pulmonary emphysema The ability of PU 163 of antagonizing the pulmonary emphysema induced in the guinea pig by intratracheal injection of 0.1 mg of porcine pancrea tic elastase has been evaluated in comparison with Ambroxol, according to the method described by Stone P.J. et al. (Am. Rev. Resp. Dis. 124/1, 5659; 1981). Both compounds have been administered by the i.p. route at the equiponderant dose of 50 mg/kg 1 hour after the elastase injection. From the results found, PU 163 turned out to be endowed with an high activity of reducing emphysema, in re markably higher extent than that found for Ambroxol. Action on the chronic bronchitis The action of PU 163 in comparison with Ambroxol on the chronic bronchitis induced by inhalation of S02 in the rat has been evaluated according to the method described by Quevauvillier e coll. (Therapie, XXII; 185-193; 1967). The compounds under exam have been administered by gastric tube at the dose of 50 mg/kg, suspended in carboxymethylcellulose. The efficacy of the tre atment has been evaluated in function of the regression degree of the pulmonary lesions induced by S02 inhalation. The results showed that both compounds exert a good activity on the pathological signs induced by the chronic bronchitis, but to a remarkably higher extent in the group of PU 163 treated animals. In fact, according to the evaluation of the lesions degrees, PU 163 showed a percent regression of 62%, compared to 38% for the Ambroxol treated group at equiponderant doses. Action on the inhibition by elastase The inhibiting activity of PU 163 and of Ambroxol on elastase has been measured according to the method described by R.M. Senior (Plenum Press. New York; 249-254; 1977). According to the results, PU 163 showed a potent elastolytic inhibiting activity with an IC50 corresponding to 11 mM/l. In this test, Ambroxol turned out to be practically inactive. Pharmacokinetics and distribution The pharmacokynetics behaviour and the distribu tion in the organs of PU 163, in comparison to Ambroxol, has been determined in the rat after ad ministration by the oral route of equimolar doses, corresponding to 50 mg/kg of Ambroxol base. The assays have been performed by HPLC and the results clearly showed some fundamental advantages of PU 163 in comparison with Ambroxol. - PU 163 can be assayed in the plasma as such in the first detection times, to give then Ambro xol; the concentrations as Ambroxol in the PU 163 treated group can be detected at prolon ged times in comparison with the Ambroxol treat ed group. This pattern of the plasma concentra tions allows a more prolonged presence in the circulation of Ambroxol, so as to allow a reduc tion of the daily administrations'frequence. - From the exam of the concentrations in the organs it was surprisingly found how the structure of PU 163 allows to carry larger amounts of Ambroxol at the pulmonary level, target organ for the thera peutic applications envisaged for the drug's use. The invention also provides pharmaceutical compositions containing, as the active ingredient, prefixed and therapeutically effective amounts of compound I or its pharmaceutically acceptable addition salts as antibronchopneumoniae agents, in admixture with a pharmaceutically acceptable carrier or diluent to be administered orally, rectally, parenteral ly or by inhalatory route. Examples of such formulations are tablets, capsu les, pills, powders, granules, lozenges, syrups, suppositories, phials, ampoules, aerosols, possible sustained release dosage forms, obtained for instan ce by microincapsulation, commonly used in pharmaceu tical technique, together with suitably excipients, disintegrating agents, tabletting aids, flavouring agents, colouring matter and other diluents as required. The preferred individual dosage unit will vary, depending upon the weight of the patient and the severity of the disease; generally it will range from 5 to 50 mg of compounds of the invention, in admixture with suitable carriers. Such dosage units will be administered from 2 to 4 times a day. CLAIMS for the Contracting States: BE, CH, DE, FR, GB, IT, LI, LU, NL, SE 1. 4-[[(2-(2-Theonyl)amino-3,5-dibromphenyl)methyl/amino/cyclohexanol of formula I EMI14.1 and its pharmaceutically acceptable acid addition salts. 2. 4-[[(2-(2-Theonyl)amino-3,5-dibromphenyl)methyl/amino/cyclohexanol hydrochloride. 3. A process for preparing compound I, which com prises reacting 4-/ /(2-amino-3,5-dibromophenyl)methyl/amino/cyclohexanol with 2-thenoyl chloride, in the presence of inert solvent and acid-binding agents. 4. A process according to claim 3, in which the reaction is effected in aromatic hydrocarbons, in the presence of alkali carbonates or bicarbonates, or tertiary amines, acting as acid-binding agents. 5. A process according to claims 3 and 4, in whi ch the reaction is effected at temperatures ranging from 10 C to the reflux temperature of the sol vent. 6. Pharmaceutical compositions having antibroncho pneumopathic activity, comprising as the active in gredient a compound of claims 1-2. 7. Pharmaceutical compositions according to claim 6, in form of capsules, tablets, suppositories, syrups, lozenges, powders, solutions.

Claims: CLAIMS for AT 1. A process for preparing compound I EMI16.1 which comprises reacting 4-/ /(2-amino-3,5-dibro mophenyl) methyl/amino/cyclohexanol with 2-thenoyl chloride, in the presence of inert solvent and acid-binding agents. 2. A process according to claim 1, in which the reaction is effected in aromatic hydrocarbons, in the presence of alkali carbonates or bicarbonates, or tertiary amines, acting as acid-binding agents. 3. A process according to claims 1 and 2, in which the reaction is effected at temperatures ranging from 100C to the reflux temperature of the solvent.

Inventors: QUADRO, GIUSEPPE
Applicants: PULITZER ITALIANA S.P.A.
Application year: 1985
Main classification: C07D333
IPC codes: C07D333