HU195836B - Process for production of derivatives of 1-n/l-/-/-4-amin-2-hydroxi-butyril/-canamicin - Google Patents

Abstract

The present invention relates to a process for the preparation of a compound of formula (I) wherein R is hydroxyl or amino, kanamycin A and canal B, and a pharmaceutically acceptable acid addition salt thereof. In the process, the kanamycin derivative of formula (II) is reacted with N-picolinoyloxy succinimide to give the 6-picolinoyl-kanamycin L-(-) - 4-picolinoylamino-2-hydroxy-butyric acid N-hydroxy-succinimide. the compound obtained is reacted with copper (II) acetate and, after separation, the resulting compound of formula (I) is recovered. J 9583G -1-

Description

The present invention relates to 1-N- (L - (-) - 4-amino-Z-hydroxybutyryl) kanamycin A and kanamycin B derivatives of formula (I): wherein, for the kanamycin A derivative, II is a hydroxyl group and kanamycin In the case of derivative B, R is an amino group and is a novel process for the preparation of acid addition salts thereof with pharmaceutically acceptable acids.

The kanamycin A derivative of formula (I), R is hydroxy, is known in the literature as nmikacin and is described in U.S. Patent 3,781,268 as having a potent antibacterial activity.

In the process of the above patent, amikacin and the corresponding kanamycin B derivative are prepared by acylating canalicin Λ or kanamycin B with benzyloxycarbonyloxysuccinimide as the acylating agent and thus at the corresponding 6-position amino group. protected kanamycin, namely 6-benzyloxycarbonyl-kanamycin.

The protected compound is then acylated with the N-hydroxysuccinimide ester of L - (-) - 4-benzyloxycarbonylamine-2-hydroxybutyric acid, and finally the reduction groups are removed by catalytic hydrogenation in the presence of a catalyst.

Italian Patent Application No. 1369,136 / 78 discloses and claims another process for aniline and its derivatives. In the process, polysilylated kanamycin A or kanamycin B is acylated to form a protecting group different from the silyl group.

In the 6-position with an acid of formula IX wherein B is a ninino protecting group and n is 0,1 or 2.

All of the protecting groups are then removed in a manner known per se, and for the carbobenzyloxy protecting group in the 6-position it is preferably carried out by catalytic hydrogenation.

Hydrogenation, which requires expensive catalysts and a suitable industrial plant, is an important operation of both known processes described above.

In addition to the above, kanamycin A and kanamycin B are used as starting materials, the preparation of the protected form requires a lengthy process and the final separation is a complex operation.

Surprisingly, a novel process for the preparation of 1-N- [L - (-) - 4-amino-2-hydroxybutyryl] -canamycin A and -canninicin B has been found which has the advantage that it does not use expensive catalysts for catalytic hydrogenation. In the process of the present invention, as described below, an easily removable protecting group is employed which results in a very good yield.

An advantage of the process of the present invention is the use of a pyridine alpha-carbonyl group (also known as a picolinoyl group) to protect the 6-position amino group and acylation with the parent canainicin1 acid. The advantage of this protecting group,. picolinic acid (pyridine-alpha-carboxylic acid) forms a stable complex with divalent copper and the resulting protected kanamycin derivative is hydrolyzed under mild reaction conditions.

The starting materials of the process of the invention, i.e., kanamycin. Antibiotics A and Kanamycin B are known in the art (Merck Index, X Ed., Pp. 758-759), which are represented by Formula II and can be prepared according to Scheme A to prepare the kanamycin derivative of Formula I.

The canainicin is dissolved in a suitable solvent, such as dimethylformamide, and then reacted with a protecting group of formula (III), namely N-picolinoyloxysuccinimide. The N-picolinoyloxy-succinimide is prepared by reacting picolinic acid or its chloride in a protic solvent preferably with hydroxysuccinimide in the presence of a dehydrating agent at room temperature or under gentle heating. The reaction is carried out cold, preferably at -10 ° C to 0 ° C, for 20-30 hours.

The 6-picolinoyl-kanamycin of the general formula (IV) thus obtained is then acylated at the 1-position. The acylation is carried out on the amino group of the N-1: hydroxy-succinimide ester of formula (VI) protected by picolinoyl-protected b - (-) - 4-amino-2-hydroxybutyric acid.

Preferably, the compound of formula (VI) is prepared so that under basic conditions, L - (-) - 4-amino--2-hydroxybutyric acid is first cold, preferably 0-5 ° C, and slowly at 30 ° C. is reacted with picolinoyl chloride. Thus, calpoll L-(-) - 4- (picolinoylamino) -2-hydroxybutyric acid (V) is prepared in an aprotic solvent such as ethyl acetal in the presence of a dehydrating agent such as cyclohexylcarbodiimide. reaction with succinimide to give the compound of formula VI.

The N-hydroxysuccinimide ester of L (-) -4- (picolinoylamino) -2-hydroxybutyric acid (VI) thus obtained is reacted with 6-picolinoyl kanamycin (IV) in a solvent at room temperature. Thus, 1-N- [L - (-) - 4-picolinoylamino-2-hydroxybutyryl] -6-picolinoyl-kanamycin (VII) is obtained directly under copper conditions under acidic conditions such as hydrochloric acid at room temperature. Treatment with (II) acetate. The copper (II) complex of formula (VIII) is obtained by protecting the amino groups with the picolinoyl group and separating the complex from the reaction mixture, thereby obtaining the 1-N- [L-2195836 (-) - 4-amino-2-hydroxy -butyryl] -canamycin, which is isolated as such or as an acid addition salt with a pharmaceutically acceptable acid.

The invention is illustrated by the following examples, without limiting the scope thereof.

Example 1 10

Preparation of N- (picoloyloxy) succininide (III) To a solution of sodium hydroxide (g, 0.22 mol) in cold water (200 mL) was added N-hydroxysuccinimide (23 g, 0.2 mol).

To the cold solution, 25.5 g (0.2 mol) of picolinoyl chloride is added dropwise.

At the end of the addition, the temperature is adjusted to θ 28-30 ° C and the solution is stirred for 12 hours.

The precipitate formed is filtered off, washed with water and recrystallized from benzene / n-hexane (10: 1) to give 37.40 g of the title compound (III) in 85% yield.

Λ The structure of the obtained compound was confirmed by analytical and spectrophotometric data.

Example 2

Preparation of N- (picoloyloxy) succininide (III)

32.4 g (0.2 mole) of N, N'-carbonyldiimidazole in solution with 200 ml of anhydrous tetrahydrofuran was treated with 24.6 g (0.2 mol) of picoline SA- ₁₀ acid.

The solution is stirred at room temperature for 1 hour or until carbon dioxide ceases.

N-hydroxysuccinimide (23 g, 0.2 mol) was then added and stirring continued for 4 hours.

The precipitated imidazole is filtered off and the tetrahydrofuran is removed under reduced pressure. The residue was crystallized from benzene / n-hexane 50 (10: 1) to give 38.5 g (87.5%) of the title compound.

the mixture was stirred for 1 hour and the temperature was slowly raised to 25-30 ° C. Ethyl acetate (100 mL) was added and the pH adjusted to about 2 with stirring.

The reaction mixture is stirred and the organic phase is separated off. The acidic aqueous solution was partitioned between ethyl acetate (50 mL). The organic phases are collected and evaporated to dryness under reduced pressure. The residue was crystallized from benzene to give the title compound (V) (16.85 g, 75%).

The structure of the resulting compound was confirmed by analytical and spectrophotometric data. ANALYSIS:

C calculated 53.35% found 54.10%

H

5:14%

5:12%

N

12:49%

12:50%

Example 4

Preparation of N-hydroxy-succinimide ester (VI) of L - (-) - 4-picolionylamino) -2-hydroxybutyric acid

11.21 g (0.2 mol) of the compound of formula V prepared in Example 3 and 5.7 g (0.5 mol) of N-hydroxysuccinimide were charged into a 500 ml flask containing 250 ml of anhydrous ethyl acetate. The reaction mixture was cooled to 0 ° C and cyclohexylcarbodimide (10.23 g, 0.5 mol) was added. Stirring is continued for 8 hours, the reaction mixture is filtered and the filtrate is concentrated to 50 ml under reduced pressure.

The title compound (VI) is filtered off in crystalline form and the crystals are washed with 25 ml of benzene and dried. Yield: 93 g (93%).

The structure of the resulting compound was confirmed by analytical and spectrophotometric data.

ANALYSIS:

C calculated 52.34% found 53%

Η N

4.71% 13.08%

4.80% 13.20%

Example 5 Preparation of d-picolinoyl-kanamycin A (IV)

Example 3

Preparation of L - (-) - 4- (picolinoylamino) -2-hydroxybutyric acid (V)

A solution of 8.387 g (0.21 mol) of sodium hydroxide in 80 ml of water was dissolved in 11.935 g (0.1 mol) of L - (-) - 4-amino-2-hydroxybutyric acid. The solution was cooled to 0-5 ° C and 12.90 g (1.01 mol) of picolinoyl chloride was added slowly with stirring over 50 minutes. THE

47 g (0.1 mol) of canainicin A (II, R = OH) are dissolved in 1 liter of a mixture of 74.4% water and 25.6% dimethylformamide.

To the solution cooled to 0 ° C, slowly, with stirring, 22 g of the

A solution of GO (0.1 mol) in III (500 ml) was added.

The reaction mixture was cooled to -10 ° C to -8 ° C and stirring was continued for 24 hours. The solution is max. Evaporate to dryness at 50 ° C.

The oily residue was taken up in water (550 mL) and n-butanol (550 mL) and the reaction mixture was stirred for 1 hour and then filtered.

Two phases are formed. The butanol solution was extracted with 2-azer 2-azer (550 mL) saturated with water and the aqueous phase was extracted 2 times with 550 mL n-butanol saturated water. The remaining butanol phase is evaporated to dryness under reduced pressure.

Some crystals form an oily residue. The crystals and oil were dissolved in methanol (100 mL), ethanol (240 mL) was added and the reaction was allowed to stand for 12 hours; the crystals thus obtained are filtered to give a mixture containing about 50 g of kanamycin A.

The crude product was dissolved in about 280 µl of cold water and the solution was adsorbed on an acidic ion exchange resin.

The column was eluted with 0.1 N ammonium hydroxide solution and the various eluted fractions obtained were subjected to chromatography.

A solution containing kanamycin A and a compound of formula IV is obtained. The fractions were evaporated to dryness under reduced pressure to give 28.1 g (about 50% yield) of compound IV and about 18 g kanamycin Λ.

The resulting compound of formula IV is used directly in the following reaction without further purification.

Example 6

Preparation of G-picolinoyl-kanamycin Β (IV)

Example 5 was repeated except that 74 g kanamycin B (II, R = NHz) was used instead of kanamycin A.

Example 7 Preliminary list of 1-N- [L - (-) - 4-picolinoylamino-2-hydroxybutyryl] -6-picolinoyl-kanamycin A (VII)

A solution of the product VI (3.21 g, 0.01 mol) in 80 ml of ethylene glycol dimethyl ether prepared in Example 4 was treated dropwise with 80 ml (5.27 g, 0.0091 mol) of the product (IV). ethylene glycol dimethyl ether and 50% water.

The reaction mixture was stirred for 12 hours at room temperature. The resulting solution was evaporated under reduced pressure to give the title compound (VII) as a dark colored material, which was used in the following procedure.

Example 8 Preparation of 1- tert- [L - (-) - 4-picolinoylamino-2-hydroxybutyryl] -6-picolionyl kanamycin Β (VII)

The procedure described in Example 7 was followed except that the derivative of the formula (IV) prepared from kanamycin B was used.

Example 9 Preparation of 1- [f - [L - (-) - 4-amino-2-hydroxybutyryl] -cannamycin A (I)

The crude product (VII) obtained in Example 7 was dissolved in 50 ml of water. To the solution was added copper (II) acetate (2.648 g, 0.012 mol) and 5% hydrochloric acid (4 ml). The reaction mixture was kept at room temperature for 2 hours and stirred occasionally. The volume was then reduced to half by evaporation under reduced pressure and the residue was filtered.

Hydrogen sulfide was bubbled through the filtrate to remove excess Cu * 'ions.

The solution was filtered again and passed through a CG-50 grade acidic resin. The resin column was then eluted with various concentrations of ammonia and the contents of the various fractions were analyzed by thin layer chromatography.

Solutions containing the product of formula (I) are collected and evaporated to dryness under reduced pressure.

Crystallization from methanol / propanol afforded the product of formula (I) in 30% yield, m.p. 203-204 ° C (hexahydrate). Yield: 22%.

[α] u ^a = 100 ° (c = 1.0 in water,

HPLC showed 95% purity.

Example 10 Preparation of 1-N- (L - (-) - 4-amino-2-hydroxybutyryl) kanamycin Β (I)

The procedure described in Example 9 was followed except that the hyamuric compound (VII) prepared from kanamycin B was used. M.p. 190-192 ° C dec. Yield: 25%

Example 11

Preparation of -N- [b - (-) - 4-amino-2-hydroxybutyryl] kanamycin A-bisulfate g amikacin base was dissolved in 20 mL water, pU was adjusted to 3 with 10% sulfuric acid and 100 mL acetone or ethanol was added to give the title compound (12.68 g).

Ik amikacin bisulfate C22H13N5 x 2H2SO4 5

High Pressure Liquid

chromatographic titer 2 72% (theoretical: 74.9%) kanamycin £ 1% Sulfate 2 23% (theoretical: 25%) Water f. 10% Loss on drying S. 10% [oc] o ”(C 1: 1120) 2 + 72 °

PATENT CLAIMS

Claims (3)

PATENT CLAIMS A process for the preparation of 1-N- [L (-) - 4-amino-2-hydroxybutyryl] canamycin A and canalaline B derivatives of formula I wherein R is hydroxy or amino, and pharmaceutically acceptable acid addition salts thereof. for the preparation of their salts by protecting the amino group at the 6-position of the kanamycin derivative (II), acylating the resulting compound, then deprotecting and optionally converting the resulting compound to an acid addition salt, characterized in that a kanamycin (II) wherein R is reacted with N-picoloyloxy-succinimide at -10 ° C to 0 ° C for 20-30 hours to give 6-picolinoyl kanamycin at room temperature, L - (-) - 4 ₃₅ picolinoyl-amino-2-hydroxybutyric acid N-hydroxysuccinimide ester is acylated, the (VII) lN- formula [L - (-) - 4-picolinoyl-amino-2-hydroxybutyryl] - 6-Picolinoyl-kanamycin is obtained in ac wherein R is as defined above, which is reacted with copper (II) acetate in an acidic medium at room temperature, then separating the divalent copper complex containing the compound of formula (I) and the protecting group of formula (VIII) and optionally compound is converted to the acid addition salt in a known manner. Process for the preparation of the N - (L - (-) - 4-amino-2-hydroxybutyryl) kanamycin derivative of the general formula (I) according to claim 1, characterized in that N- picolionyl oxysuccinimide is prepared by reacting the picolinic acid or its chloride with hydroxyazuccinimide, preferably in the presence of a dehydrating agent in a protic solvent, at room temperature or with slight heating. 3. A process according to claim 1 for the preparation of the N - (L - (-) - 4-amino-2-hydroxybutyryl) kanamycin derivative of the formula (I), wherein -picolinoylamino-2-hydroxybutyric acid N-hydroxysuccinimide ester, used to treat L - (-) - 4-amino-2-hydroxybutyric acid with piconoyl chloride in basic medium at 0 to 30 ° C. and reacting the resulting L - (-) - 4-picolionylamino-2-hydroxybutyric acid with N-hydroxysuccinimide in an aprotic solvent in the presence of a dehydrating agent.