JPS5910360B2 - Improved production method of 1-N-[L-(1)-γ-amino-α-hydroxybutyryl]-kanamycin derivative - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an improved process for the production of 1-N-[L-(-)-γ-amino-α-hydroxybutyryl]monokanamycin A with substantially increased efficiency.

The present invention relates to the production of 1-N-[L-(-)-γ-amino-α-hydroxybutyryl]-kanamycin A (■) (6'-N-(benzyloxycarbonyl)kanamycin A () in a solvent). 1 mole of I) is treated with at least 3 moles of aromatic or aliphatic aldehyde to produce 6'-N-(benzyloxycarbonyl)kanamycin A Schiff base (■) (0-5 in a solvent).
At a temperature in the range of 0°C, 1 mol of compound (■)
In the presence of a catalytic amount of 1-hydroxybenztriazole,
0.3 to 1.5 mol of synchhexylcarpodiimide or 1-ethyl-3-(N-N-dimethylaminopropyl)-carbodiimide can be added to 0.3 to 1.5 mol of L-(1)-γ-(benzyl oxycarbonyl)amino-α-hydroxybutyric acid and then treated with acid to give 1-N-[L-(-)--(benzyloxycarbonyl)amino-α-hydroxybutyryl]-6'-N
-(benzyloxycarbonyl)kanamycin A (■)
(1-N-[L-(1)-γ) characterized by a continuous process of producing compound () by hydrogenolyzing compound () with hydrogen in a solvent in the presence of a metal catalyst. -Amino-
This invention relates to an improved method for producing α-hydroxybutyryl]-kanamycin A.

The Schiff base referred to in the present invention is a triarylidene Schiff base.

Amikacin (AlllalLCin), which is one of the 1-N-[L-(-)-γ-amino-α-hydroxybutyryl] monokanamycin derivatives, is a semisynthetic derivative of kanamycin A, and is particularly effective against kanamycin-resistant bacteria. It is important as a drug that shows efficacy.

Kanamycin A has molecules 1, 3, 6' and ? It has four amino groups in the positions. When these amino groups are reacted with an acylating agent, it is known that the 6'-amino group has the highest reactivity, and the 1-amino group has the second highest reactivity.
It has been confirmed that both 3- and 3″-amino groups are less reactive than either 1- or 6′-amino groups (Literature; Kawaguchi, Naito, Nakagawa, Fujisawa; Journal of Antibiotics, Vol. Volume 25, pages 695-708,
(1972). Therefore, when amikacin is synthesized using kanamycin A as a starting material, the 6' of kanamycin A is
-Protect the amino group with a suitable protecting group, followed by L-(-)
Amikacin is chemically synthesized by reacting an active derivative of the carboxyl group of -r-monoprotected amino-α-hydroxybutyric acid. 6'-N-acyl kanamycin derivatives have been prepared by a method already pending by the present inventors (Japanese Patent Application No. 1973).
-127133).

Since the active derivative of L-(-)-r monoprotected amino-1α-hydroxybutyric acid with respect to the carboxyl group has strong reactivity with amino groups, it was allowed to act on the 65-N-acyl kanamycin derivative. In this case, the position where L-(-)-r monoprotected amino monoα-hydroxybutyric acid is introduced is
Although the 1-amino group is primary, the 3- and ? - also reacts with amino groups to produce undesirable acylation products.

The formation of these undesirable acylated products significantly reduces the efficiency of amikacin production and is extremely disadvantageous for the industrial production of amikacin. Therefore, the present inventors and co-researchers have developed a method that aims to selectively acylate the 1-amino group over other amino groups in the molecule (Japanese Patent Application No. 1971-69188, JP-A-48-34856). Alternatively, a method aiming at improving efficiency by converting the amino group in the molecule into a Schiff base (Japanese Patent Application No. 1986-1)
No. 1524, Japanese Unexamined Patent Publication No. 49-102644) was proposed, but it cannot be said that the method is necessarily satisfactory.

The present inventor conducted various studies and examinations with the aim of improving the production efficiency of 1-N-[L-(-)-γ-amino-α-hydroxybutyryl] monokanamycin derivatives, and found that ( A) Treating 1 mol of compound (1) with at least 3 mol of aromatic aldehyde in a solvent to form compound ()
(B) 0.3 to 1.5 mol of synchlorohexylcarbodiimide in the presence of a catalytic amount of 1-hydroxybenztriazole to 1 mol of compound () in a solvent at a temperature in the range of 0 to 50°C. or 1-ethyl-3-(N
-N-dimethylaminopropyl)-carbodiimide with 0.3 to 1.5 mol of L-(-)-r-(benzyloxycarbonyl)amino-d-hydroxybutyric acid, and then treated with acid. 1-N-[L-(- The present inventors have discovered that the process for producing γ-amino-α-hydroxybutyryl]-kanamycin A () achieves the object of the present invention, and has completed the present invention.

The method of the present invention includes water, acetone, tetrahydrofuran,
in a solvent selected from the group consisting of dimethylformamide, dimethylacetamide, acetonitrile, dioxane, pyridine, propylene glycol dimethyl ether, ethylene glycol dimethyl ether or a mixed solvent thereof at a temperature in the range of 0 to 50 °C, preferably 15 to 25 °C.
0.3 to 1 mole of compound () in the presence of a catalytic amount of 1-hydroxybenztriazole, preferably from 0.01 to 1 mole, most preferably from 0.05 to 0.2 mole, at °C.
~1.5 mol, preferably 0.5 to 1 mol of synchhexylcarbodiimide or 1-ethyl-3-(N-N
-dimethylaminopropyl)-carbodiimide,
0.3 to 1.5 mol, preferably 0.5 to 1 mol of L-(
-)-r-Acylamino-d-hydroxybutyric acid is reacted and then hydrolyzed with a mineral acid.

Furthermore, the method of the present invention comprises (A) a solvent selected from the group consisting of water, methanol, ethanol, propanol, acetone, methyl ethyl ketone, methyl isobutyl ketone, ethyl acetate, butyl acetate, benzene, diethyl ether, methylene chloride, chloroform, or a mixed solvent thereof; in the selected solvent, preferably a mixed solvent of water and methanol or a mixed solvent of water and methyl isobutyl ketone, at a temperature in the range of 0 to 100 °C,
Preferably at 20-50°C, for 1-10 hours, 1 mol of compound (1) is added to 3-6 mol, preferably 3-5 mol of an aromatic aldehyde, preferably benzaldehyde, hydroxybenzaldehyde, methoxybenzaldehyde or Compound () treated with halogenobenzaldehyde
(8) water, acetone, tetrahydrofuran,
in a solvent selected from the group consisting of dimethylformamide, dimethylformamide, acetonitrile, dioxane, pyridine, propylene glycol dimethyl ether, ethylene glycol dimethyl ether or a mixed solvent thereof at a temperature in the range of 0 to 50 °C, preferably 15 to 25 °C.
0.3 to 1 mole of compound () in the presence of a catalytic amount of 1-hydroxybenztriazole, preferably from 0.01 to 1 mole, most preferably from 0.05 to 0.2 mole, at °C.
~1.5 mol, preferably 0.5 to 1 mol of synchhexylcarbodiimide or 1-ethyl-3-(N-N
-dimethylaminopropyl)-carbodiimide,
0.3 to 1.5 mol, preferably 0.5 to 1 mol of L-(
-)-r-(benzyloxycarbonyl)amino-α-
Hydroxybutyric acid is reacted and then hydrolyzed with mineral acid to produce the compound () (Q water and tetrahydrofuran, methanol, ethanol, propanol, dimethylformamide, dimethylacetamide, dioxane, propylene glycol dimethyl ether, and ethylene glycol). Hydrogen in the presence of a metal catalyst selected from the group consisting of palladium, platinum, rhodium, Raney nickel, ruthenium and nickel, preferably palladium on carbon, in a water-water miscible solvent system selected from the group consisting of dimethyl ether. This is an improved method for producing 1-N-[L-(-)-r-amino-d-hydroxybutyryl]-kanamycin A, which is characterized by a continuous process of producing compound () by hydrogenolysis.

The 6'-N-acyl kanamycin derivatives used in the method of the present invention include 6'-N-acyl kanamycin A and 6
'-N-acylkanamycin B may be mentioned.

A reactant obtained according to the process of the invention, for example 6'-N-acylkanamycin Schiff base () in aqueous tetrahydrofuran, is added to L-(-)-1 in the presence of a catalytic amount of 1-hydroxybenztriazole. r-acylamino-α-
The reaction product obtained by treatment with hydroxybutyric acid and synchlohexylcarbodiimide followed by hydrolysis with mineral acid is a mixture of compound (), positional isomer of compound (), polyacylated product, and unreacted compound (1). However, these mixtures can be prepared using a method that the present inventors have already applied for (Japanese Unexamined Patent Publication No. 127951/1971, December 7, 1972), that is, synthetic adsorption resins (for example, Mitsubishi High Porous Polymer HP-
208) can be used to separate them from each other.

Another method for separating the reaction mixture is to use an ion exchange resin, but this method is not particularly advantageous. The separated unreacted compound (1) is 1-N-[L-(1)-r
-amino-α-hydroxybutyryl] can be recycled and used in the next reaction as an intermediate raw material for the synthesis of kanamycin derivatives. The separated isomers and polyacylated products were obtained using a method already applied by the present inventors (Japanese Patent Application No. 4).
No. 9-022369 (Japanese Unexamined Patent Publication No. 112334/1989))
The acyl group can be removed by alkaline hydrolysis and returned to biologically active kanamycin A or kanamycin B.
It can be used as a raw material for the synthesis of hydroxybutyryl]-kanamycin derivatives. The method of the present invention is 1-N-[
L-(-)-γ-amino-α-hydroxybutyryl] is included in the reactant obtained by the method of the present invention when producing amikacin, for the purpose of demonstrating the fact that it improves the efficiency of the production of kanamycin derivatives. Each component is separated using Mitsubishi High Porous Polymer HP-208, and each separated component is then treated with amikacin (base) unreacted compound ().
The recovered kanamycin monosulfate A was collected, and the yields were as shown in Table 1.

As is clear from the results in Table 1, the method of the present invention applies L-(
-)-γ-Amino-α-hydroxybutyric acid was introduced in a significantly large amount, resulting in 1-N-[L-(-)-r-amino-
It has been demonstrated that the efficiency of the production of α-hydroxybutyryl]-kanamycin derivatives has been significantly improved.

Furthermore, according to the method of the present invention, a considerable amount of unreacted compound ( ) and kanamycin are recovered, so that 1-N-[L-
(-)-r-amino-d-hydroxybutyryl] monokanamycin derivatives has also been demonstrated to be useful in improving the efficiency of production. Next, the method of the present invention will be explained in detail with reference to Examples, but the present invention is not limited thereto.

Reference Example 6'-N-penzyloxycarbonyllucanamycin A
Production of kanamycin monosulfate A253f (kanamycin A
O. Kanamycin A was dissolved as the free base by suspending 4l3 mol) in water 11 and adding triethylamine 120a to it.

Para(benzyloxycarbonyloxy)benzoic acid 84
y (0.31 mol) was slowly added to the kanamycin solution over a period of 30 minutes, during which time 60 ml of triethylamine
C-PH was maintained at 9.8 to 10.0 and the temperature was maintained at 20 to 25°C while stirring. After further stirring at room temperature for 2 hours, the reaction solution was left at room temperature overnight and the pH was adjusted to 9 with dilute sulfuric acid, and methanol 21 and seed crystals were added to adjust the pH to 30-35.
By gently stirring the mixture at ℃ for 30 minutes, crystals of kanamycin A monosulfate were precipitated.

Subsequently, 0.651 g of methanol was added, and the mixture was slowly stirred at 20 to 24° C. for 5 hours to effect sufficient crystallization, and the mixture was further left at room temperature overnight.

The precipitated crystals of kanamycin monosulfate A were separated from each other, and the cake was washed with 75% methanol water.

Combine the water-containing methanol F solution and the washing solution, and add 65-N-
3.81 g of a hydrous methanol solution of benzyloxycarbonyl-kanamycin A was obtained.

Example 16'-benzyloxycarbonyl-1●3.
Production of 3I-trisalicylidene-kanamycin A 1.91 of the hydrous methanol solution of 6'-N-benzyloxycarbonyl-kanamycin A obtained in Reference Example was corrected to pH 8.2 with dilute sulfuric acid, and then at room temperature. Add 60ml of salicylaldehyde, add seed crystals,
Stirring was carried out for an hour.

It was left at room temperature overnight. The precipitated pale yellow crystals were collected, and the cake was washed with 75% methanol water and dried with warm air at 50 to 55°C, resulting in 6'-benzyloxycarbonyl-1,3,3''-trisalicylidene-kanamycin. Al2O7 was obtained.

Melting point: 177-180°C, specific rotation: [d] Blue°C +25
．． 87 (Cl, dioxane) Example 2 6'-benzyloxycarbonyl-1.3.? Production of Tribenzylidene-Kanamycin A 1.91 g of the water-containing methanol solution of 6'-N-benzyloxycarbonyl-kanamycin A obtained in Reference Example was evaporated under reduced pressure to remove methanol.

The residue was diluted with water to a total volume of 2.51 ml, 0.51 ml of methyl isobutyl ketone was added, and 60 ml of benzaldehyde was added.
1 was added and stirred vigorously at room temperature for 3 hours. The precipitated white precipitate was collected, and the cake was washed with methyl isobutyl ketone and water, and then dried with warm air at 50 to 55°C. As a result, 6'-benzyloxycarbonyl-1.3.? -tribenzylidene-kanamycin All
I got the OV. Decomposition point: 194-198°C, Specific rotation: [α] Sweet°C + 35: (Cll methanol), Total nitrogen:
6.1% Example 3 Production of amikacin 6'-benzyloxycarbonyl-1,3,j-trisalicylidene-kanamycin AlOOf7 (0.1074
1.140 ml of tetrahydrofuran and 80 ml of water
It was dissolved in ml of mixed solvent.

Subsequently, 27.2 f (0.1074 mol) of L-(-)-γ-benzyloxycarbonylamino-α-hydroxybutyric acid and 1.35 t of 1-hydroxybenztriazole were added and dissolved to form a homogeneous solution. A solution of 24.3 t (0.118 mol) of synchhexylcarbodiimide dissolved in 300 ml of tetrahydrofuran was added to this at 20°C.
The mixture was added slowly over 6.5 hours with vigorous stirring. Stirring was continued for an additional 16 hours at 20°C. 500 ml of water was added to the reaction solution, and the PHL. 5 and stirring for 1 hour to perform acid hydrolysis.
After separating the precipitated crystals (synchrohexyl urea),
Tetrahydrofuran in the slurry was distilled off by evaporation under reduced pressure. The crystals (synchrohexyl urea) precipitated again in the residue were separated, and the crystal cake was washed with water and methyl isobutyl ketone. Combine the wash liquor, water washing and methyl isobutyl ketone washing, and add 30 ml of methyl isobutyl ketone.
After adding 0 ml of the mixture, the mixture was stirred at room temperature for 30 minutes. The mixture was separated into an aqueous layer and an organic solvent layer by liquid separation, methyl isobutyl ketone dissolved in the aqueous layer was removed by evaporation under reduced pressure, and the residue was diluted with water to give a total volume of 2000 d. After removing the precipitate (a small amount), the slurry was lowered to a tower filled with Mitsubishi High Porous Polymer HP-2082.51 at pH 2.
The liquid was passed through at a flow rate of Sv=1, and the column was subsequently washed with 6000 ml of water. Unreacted 6''-benzyloxycarbonyl-kanamycin A was found in the column flow and washing liquid, so this fraction was collected and recovered as Schiff's base.
-benzyloxycarbonyl-1.3.? Monotrisalicylidene-kanamycin Al 8.27 (18.2%) was obtained. The reaction product adsorbed in the column is charged with 0.02N hydrochloric acid 37.5% methanol aqueous solution 7.51.0.
02 Normal hydrochloric acidic 60% methanol aqueous solution 51 and 0
．． It is eluted by sequentially flowing 3.5 liters of 2N hydrochloric acid acidic methanol solution. Compound () was observed in the section eluted with 37.5% methanol aqueous solution.

The positional isomer of compound () was found in the fraction eluted with 60% methanol aqueous solution, and the polyacylated product was found in the fraction eluted with methanol, so these fractions were combined, and after neutralization with aqueous ammonia, Methanol was removed by evaporation under reduced pressure, and the residue was further concentrated in vacuo, and then sodium hydroxide was added to make a 4N sodium hydroxide solution, followed by alkaline hydrolysis by continuing heating at 80 to 95° C. for 3 hours.

By this treatment, the positional isomer and polyacylated product of compound () were returned to kanamycin A, so after neutralizing the aqueous solution, the conventional kanamycin A collection process was carried out to obtain kanamycin monosulfate (crystal) 17r (26.4 %) were recovered. The 37.5% methanol-water eluted fraction containing compound () was adjusted to pH 3.5 with aqueous ammonia, methanol was removed by evaporation under reduced pressure, and further concentrated in vacuo to obtain 250 WLI of a concentrated solution.

To this was added 250 TfLe of methanol, 5 tons of 10% palladium on carbon, and the mixture was stirred for 16 hours under a hydrogen stream to perform hydrolysis. After removing methanol from the hydrolysis solution by evaporation under reduced pressure, water was added to the residue and 3
An aqueous solution (PH6.8) of 00d was prepared. Add this solution to Amberlite CG-508 (ammonium type) 300
Amikacin was adsorbed onto the resin by flowing it into the column d.

After washing the column with water (900 TIL1) and 0.3 N ammonia aqueous solution (1200 m0), amikacin is eluted by flowing a 0.5 N ammonia aqueous solution (3000 d) in a descending manner at SV = 0.5. The amikacin aqueous solution was concentrated in vacuo to form a syrup. About 4 times the volume of methanol was added to this to crystallize amikacin (base). The crystal cake was removed by filtration, washed with 80% methanol water, and concentrated in vacuo. When dried, amikacin 22.7f was obtained as a white crystalline powder.Bioassay 880 μ7 titer/T!9 (yield 31.8%) Moisture measurement (Karl Fischer method) 6.0% Example 4 Preparation of amikacin 6'-benzyloxycarbonyl-1,3-?-trisalicylidene-kanamycin AlOOV (0.1074 mol) was mixed with 1140 d of tetrahydrofuran and 80 WLt of water.
L-(-)-γ-benzyloxycarbonylamino-α-hydroxybutyric acid 19f (
0.075 mol) and 1 V of 1-hydroxybenztriazole were added and dissolved to form a homogeneous solution.

Synchhexylcarbodiimide 17f7 (0.082
A solution prepared by dissolving 5 moles of 5 moles in 300 ml of tetrahydrofuran was slowly added to the homogeneous solution at 20° C. over 3 hours with vigorous stirring. Another 20-3
Stirring was continued for 16 hours at 0°C. The reaction solution was treated and separated and purified in the same manner as in the middle and lower rows of Example 3 to obtain 18.75 t of amikacin as a white crystalline powder from the compound ().

Bioassay 897μr titer/η (yield 26.7%) Moisture measurement (Karl Fischer method) From the 6.0% unreacted fraction, recovered as -Schiff base, 6'-benzyloxycarbonyl-1.3. 3''-trisalicylidene-kanamycin A35.2f7 (yield 35.2%) was obtained.

Kanamycin monosulfate AlOf7 (15%) was recovered from the positional isomer and polyacylated fractions of compound ().

Example 5 Production of amikacin 6'-benzyloxycarbonyl-1,3,3''-tribenzylidene-kanamycin A88.2f7 (0.1 mol) was dissolved in a mixed solvent of 900 ml of tetrahydrofuran and 45 d of water, and then L-(1)-r-benzyloxycarbonylamino-d-hydroxybutyric acid 25.3y(
0.1 mol) and 1-hydroxybenztriazole 1.
5t was added and dissolved to form a homogeneous solution.

A solution of 20.6f (0.1 mol) of synchhexylcarbodiimide dissolved in 200d of tetrahydrofuran was slowly added to the above homogeneous solution over 5 hours at 10-15°C with vigorous stirring.

Stirring was continued for further 16 hours at 20-30°C. The reaction solution was treated, separated and purified in the same manner as in the middle and lower rows of Example 3 to obtain amikacin 20.3r as a white crystalline powder from the compound ().

Bioassay 886μt titer/7V (yield 30.8%) Moisture measurement (Karl Fischer method) 6.6% Recovered as Schiff's base from the unreacted fraction Tribenzylidene-kanamycin A22t (25%) was obtained.

Kanamycin monosulfate Al3.7f7 (kanamycin A
lO. 75f; recovery rate 22.2%) was recovered.

Example 6 Preparation of amikacin 6'-benzyloxycarbonyl-1,3,3''-tribenzylidene-kanamycin A88.27 (0.1 mol) was mixed with 925 ml of tetrahydrofum and 507.12 ml of water.
and then L-(-)-1γ-benzyloxycarbonylamino-α-hydroxybutyric acid 22.8
t (0.09 mol) and 1-hydroxybenztriazole 1f were added and dissolved to form a homogeneous solution.

Synchhexylcarbodiimide 18.5t (0.09
200 ml of a solution of mol) in tetrahydrofuran was slowly added to the homogeneous solution over 4 hours at 15-20° C. with vigorous stirring.

Stirring was continued for further 24 hours at 25-30°C.

The reaction solution was treated, separated and purified in the same manner as in the middle and lower rows of Example 3 to obtain Amikacin 18t as a white crystalline powder from the compound (). Bioassay 9
04μ7 titer/η (yield 28%) Moisture measurement (Karl Fischer method) 6'-benzyloxycarbonyl-1.
3.? Tribenzylidene-kanamycin A267 (28
%) was obtained.

Kanamycin monosulfate Al4. l5f7 (23%) was recovered.

Claims (1)

[Scope of Claims] 1 In the production of 1-N-[L-(-)-γ-amino-α-hydroxybutyryl]-kanamycin A (IV) (
A) 6'-N-(benzyloxycarbonyl) in solvent
One mole of kanamycin A (I) is treated with at least 3 moles of aromatic aldehyde to form 6'-N-(benzyloxycarbonyl)-kanamycin A Schiff base (II).
(Sits base means -1,3,3''-tri-benzylidene Schiff base and -1,3,3''-tri-salicylidene Schiff base), (B) in a solvent, 0 to 5
To 1 mole of compound (II) at a temperature in the range of 0° C., in the presence of a catalytic amount of 1-hydroxybenztriazole, 0° C.
．． 0.3 to 1.5 mol of L-(-)-γ-(benzyloxycarbonyl ) amino-α-hydroxybutyryl and then treated with acid to give 1-N-[L-(-)-γ-(benzyloxycarbonyl)amino-α-hydroxybutyryl]-6′-N
-(benzyloxycarbonyl)kanamycin A (III
), then (C) in a solvent in the presence of a metal catalyst,
Compound (III) is hydrolyzed with hydrogen to produce compound (IV
) characterized by a continuous process for producing 1-N-[L-(
An improved method for producing -)-γ-amino-α-hydroxybutyryl]-kanamycin A.