METAL COMPLEXES OF N-METHYL-11-AZA-10-DEOXO-10-DIHYDRO-ERYTHROMYCIN A OR 11-AZA-10-DEOXO-10-DIHYDROERYTHROMYCIN A, METHOD FOR THE MANUFACTURE THEREOF AND THEIR USE IN THE MANUFACTURE OF PHARMACEUTICALS

Abstract: New 2:1 complexes of N-methyl-11-aza-10-deoxo-10-dihydro­ erythromycin A or 11-aza-10-deoxo-10-dihydroerythromycin A with bivalent metals chosen from the group comprising Cu⁺², Zn⁺², Co⁺², Ni⁺² and Ca⁺², a process for their manufacture and their use in the manufacture of antibiotically active pharmaceuticals. New 2:1 complexes of N-methyl-11-aza-10-deoxo-10-dihydro­ erythromycin A or 11-aza-10-deoxo-10-dihydroerythromycin A with bivalent metals chosen from the group comprising Cu⁺², Zn⁺², Co⁺², Ni⁺² and Ca⁺² are made by reacting N-methyl-11-aza-­ 10-deoxo-10-dihydroerythromycin A or 11-aza-10-deoxo-10-dihydro­ erythromycin A in its free base form or in its water-soluble salt form with salts of said bivalent metals. The compounds possess antibiotic activity.

Description: The present invention relates to new, biologically active compounds of semisynthetic 15-membered macrolide antibiotics N-methyl-11-aza-10-deoxo-10-dihydroerythromycin A or 11-aza-10-deoxo-10-dihydroerythromycin A with bivalent metals, to methods for the manufacture thereof and to their use in the manufacture of pharmaceuticals. By means of chemical transformations of the C-9 keto group of erythromycin A there were synthetized 15-membered macrolides possessing a nitrogen atom in the aglycone ring, namely 11-aza-10-deoxo-10-dihydroerythromycin A (U.S. patent 4,328,334) and N-methyl-11-aza-10-deoxo-10-dihydroerythomycin A (Belgian patent 892,357 and GB patent 2,094,293 resp.), which is an effective antibacterial agent in the treatment of gram-positive and gram-negative microorganisms and exhibited an important activity in preliminary in vivo tests. It has been known that the presence of metals and the formation of metal complexes resp. might substantially influence the stability, distribution, biotransformation, elimination and other characteristics of pharmaceuticals, especially of antibiotics. According to the literature(J. Pharm. Pharmac. 18 (1966) 729) erythromycin A may theoretically yield a weak complex with Co⁺², but it does not yield any complexes with Cu⁺², Ca⁺², Mg⁺², Ni⁺² and Zn⁺² ions. The metal complexes of N-methyl-11-aza-10-deoxo-10-dihydroerythromycin A and 11-aza-10-deoxo-10-dihydroerythromycin A have, according to the Applicant's own search of prior art, not been described as yet. It has now been found that new useful complexes of N-methyl-11-aza-10-deoxo-10-dihydroerythromycin A (Ia) and 11-aza-10-deoxo-10-dihydroerythromycin A (Ib) with bivalent metals chosen from the group comprising Cu⁺², Zn⁺², Co⁺², Ni⁺² and Ca⁺² in the ratio of 2:1 may be obtained in a simple manner and in high yields by means of the claimed process. One object of the present invention are new 2:1 complexes of N-methyl-11-aza-10-deoxo-10-dihydroerythromycin A and 11-aza-10-deoxo-10-dihydroerythromycin A of the above cited formulae Ia and Ib with Cu⁺², Zn⁺², Co⁺², Ni⁺² and Ca⁺², which exhibit a superior pharmacological, specifically a superior antibiotic activity. Another object of the present invention is a new process for the manufacture of said new metal complexes, which is performed in such a manner that N-methyl-11-aza-10-deoxo-10-dihydroerythromycin A of the formula Ia or 11-aza-10-deoxo-10-dihydroerythromycin A of the formula Ib or salts thereof are reacted with salts of bivalent metals chosen from the group comprising Cu⁺², Zn⁺², Co⁺², Ni⁺² and Ca⁺², in a molar ratio of 2:1, at ambient temperature. When the reaction is performed with the compounds Ia or Ib in their salt form, said compounds Ia or Ib are first converted into their water-soluble salts, e.g. a hydrochloride, by means of suitable acids, whereupon the reaction is performed in aqueous solution with a salt of said bivalent metals. The pH of the reaction solution is adjusted to 8-11 by adding alkali lyes (pH-stating) and the product is isolated by means of filtration (method A). When the reactants Ia and Ib are used in their free base form, the reaction with the metal salt is performed in an alcohol-water mixture while adjusting the pH value by means of alkali lyes and evaporating the alcohol under reduced pressure. The product is isolated by filtration (method B). The antibiotical activity was assessed on the test organism Sarcina lutea ATCC 9341 as evident from the following Examples. The new complexes exhibited a potent antibiotic activity. Therefore a further object of the present invention is a pharmaceutical composition comprising an effective amount of the new metal complexes of N-methyl-11-aza-10-deoxo-10-dihydroerythromycin A or 11-aza-10-deoxo-10-dihydroerythromycin A, a method of treating human and animal infections, and a method for the manufacture of pharmaceuticals comprising the metal complexes of the compounds of the formula Ia or Ib. The present invention is illustrated but in no way limited by the following Examples. Example 1 (Method A) 0.749 g N-methyl-11-aza-10-deoxo-10-dihydroerythromycin A were charged ino a 100 ml beaker and dissolved under stirring and the addition of 1 N HCl in 50 ml of water (0.02 M solution) (pH about 6). Then there were added 0.086 g CuCl₂ x 2H₂O (0.01 M solution with respect to Cu⁺²) and the stirring was continued under a stepwise addition of 0.1 N NaOH until the pH value of 8.5 was achieved. Subsequently, the reaction mixture was stirred for 2 hours at a constant pH value (pH-stating), then the violet-coloured precipitate was aspirated, washed three times with 10 ml of water and dried; and yield was 0.64 g of the product (81.8 %). Cu-analysis: (polarographic method, 0.1 N HCl, E 1/2 = -0.25 V; SCE - saturated calomel electrode) Calc.: 4.07 % Found: 4.1 % Activity: 834 U/mg Sarcina lutea ATCC 9341. Example 2 (Method B) In 50 ml of a 0.02 M solution of N-methyl-11-aza-10-deoxo-10-dihydroerythromycin A in 60 % w/w methanol there were dissolved 0.086 g CuCl₂ x 2H₂O (0.01 M solution with respect to Cu⁺²). After the adjustment of the pH value to 8.5 with 0.1 N NaOH it was kept stirring for two hours at ambient temperature. The reaction mixture was evaporated at reduced pressure to about half its volume, the violet-coloured precipitate was aspirated, washed threee times with 10 ml of water, dried and afforded 0.62 g of the product (79.3 %). The analysis of the product was identical as in Example 1. Example 3 The process was performed as described in Example 1 with the sole exception that 0.068 g ZnCl₂ were added instead of CuCl₂ and the pH value was kept adjusted at 8.6. There were obtained 0.61 g of a white-coloured product (77.9 %). Zn-analysis: (atom absorption spectrophotometric method) Calc.: 4.18 % Found: 4.5 % Activity: 852 U/mg Sarcina lutea ATCC 9341. Example 4 The process was performed as described in Example 1 with the sole exception that 0.118 g CoCl₂ x 6H₂O were added instead of CuCl₂ and the pH value was kept adjusted at 8.6. There were obtained 0.63 g of a light green-coloured product (80.7 %). Co-analysis: (polarographic method, 0.1 N HCl - 0.1 N KCl (1:25), E 1/2 = -1.60 V; SCE) Calc.: 3.79 % Found: 4.1 % Activity: 849 U/mg Sarcina lutea ATCC 9341. Example 5 The process was performed as described in Example 1 with the sole exception that 0.119 g NiCl₂ x 6H₂O were added instead of CuCl₂ and the pH value was kept adjusted at 8.6. There were obtained 0.62 g (79.6 %) of a light green-coloured product. Ni-analysis: (polarographic method, 0.1 N HCl - 0.1 N KCl (1:25), E 1/2 = -1,55 V; SCE) Calc.: 3.77 % Found: 4.3 % Activity: 852 U/mg Sarcina lutea ATCC 9341. Example 6 The process was performed as described in Example 1 with the sole exception that 0.074 g CaCl₂ x 2H₂O were added instead of CuCl₂ and the pH value was kept adjusted at 8.6. There were obtained 0.60 g (77.9 %) of a white-coloured product. Ca-analysis: (atom absorption spectrophotometric method) Calc.: 2.60 % Found: 3.3 % Activity: 856 U/mg Sarcina lutea ATCC 9341. Example 7 The process was performed as described in Example 1 with the sole exception that 0.735 g 11-aza-10-deoxo-10-dihydroerythromycin A were charged instead of N-methyl-11-aza-10-deoxo-10-dihydroerythromycin A and the pH value was kept adjusted at 9.0. There were isolated 0.62 g of a violet-coloured product (80.8 %). Cu-analysis (polarographic method: 0.1 N HCl) Calc.: 4.14 % Found: 4.4 % Activity: 945 U/mg Sarcina lutea; ATCC 9341. Example 8 The process was performed as described in Example 2 except that there were charged 0.735 g 11-aza-10-deoxo-10-dihydroerythromycin A instead of N-methyl-11-aza-10-deoxo-10-dihydroerythromycin A and 0.068 g ZnCl₂ instead of CuCl₂, while keeping the pH value adjusted at 10.0 for three hours. There were isolated 0.53 g (69.0 %) of a white-coloured product. Zn-analysis: (atom absorption spectrophotometric method) Calc.: 4.10 % Found: 4.6 % Activity: 530 U/mg Sarcina lutea ATCC 9341. Example 9 The process was performed as described in Example 4 except that 0.735 g 11-aza-10-deoxo-10-dihydroerythromycin A were charged instead of N-methyl-11-aza-10-deoxo-10-dihydroerythromycin A and the pH value was kept adjusted at 10. There were isolated 0.54 g (70.6 %) of a light green-coloured product. Co-Analysis: (polarographic method, 0.1 N HCl - 0.1 N KCl) Calc.: 3.72 % Found: 4.1 % Activity: 435 U/mg Sarcina lutea ATCC 9341. Example 10 The process was performed as described in Example 9 with the sole exception that 0.118 g NiCl₂ x 6H₂O were charged instead of CoCl₂. There were isolated 0.56 g (73.2 %) of a light green-coloured product. Ni-Analysis: (polarographic method, 0.1 N HCl - 0.1 N KCl) Calc.: 3.70 % Found: 4.1 % Activity: 500 U/mg Sarcina lutea ATCC 9341. Example 11 The process was performed as described in Example 9 with the sole exception that 0.074 g CaCl₂ x 2H₂O were added instead of CoCl₂. There were isolated 0.52 g (68.9 %) of a white-coloured product. Ca-analysis: (atom absorption spectrophotometric method) Calc.: 2.55 % Found: 3.0 % Activity: 517 U/mg Sarcina lutea ATCC 9341.

Claims: A complex of N-methyl-11-aza-10-deoxo-10-dihydroerythromycin A of the formula (Ia) or 11-aza-10-deoxo-10-dihydroerythromycin A of the formula (Ib) with bivalent metals chosen from the group comprising Cu⁺², Zn⁺², Co⁺², Ni⁺², Ca⁺² in the ratio of 2:1. A process for preparing a complex of N-methyl-11-aza-10-deoxo-10-dihydroerythromycin A (Ia) or 11-aza-10-deoxo-10-dihydroerythromycin A (Ib) with bivalent metals chosen from the group comprising Cu⁺², Zn⁺², Co⁺², Ni⁺² and Ca⁺² in the ratio of 2:1, which comprises: A) the reaction of a compound of the formula (Ia) or (Ib) in the form of its water-soluble salt with a salt of a bivalent metal chosen from the group comprising Cu⁺², Zn⁺², Co⁺², Ni⁺² and Ca⁺², in a molar ratio of 2:1, in an aqueous solution and at ambient temperature while keeping the pH value adjusted at 8-11, and the isolation of the product by means of filtration, or B) the reaction of a compound of the formula (Ia) or (Ib) with a salt of a bivalent metal chosen form the group comprising Cu⁺², Zn⁺², Co⁺², Ni⁺² and Ca⁺², in a molar ratio of 2:1, at ambient temperature, in an alcohol-aqueous solution while keeping the pH value adjusted at 8-11 by the addition of alkali lyes, the evaporation of the alcohol under reduced pressure after the performed reaction and the isolation of the product by filtration. A process as claimed in claim 2, wherein the water-soluble salt of the compound (Ia) or (Ib) is a hydrochloride. A process as claimed in claims 2 or 3, wherein the metal salt is a chloride. The use of 2:1 complexes of N-methyl-11-aza-10-deoxo-10-dihydroerythromycin A or 11-aza-10-deoxo-10-dihydroerythromycin A with bivalent metals chosen from the group comprising Cu⁺², Zn⁺², Co⁺², Ni⁺² and Ca⁺² in the manufacture of antibiotically active pharmaceuticals.

Inventors: DJOKIC, SLOBODAN, DR., LOPOTAR, NEVENKA, VAJTNER, ZLATKO, DR.
Applicants: SOUR PLIVA FARMACEUTSKA, KEMIJSKA PREHRAMBENA I KOZMETICKA INDUSTRIJA, N.SOL.O.
Application year: 1987
Main classification: C07H17
IPC codes: A61K31, C07H17, A61P31, C07F1