KR890002889B1 - Process for the preparation of amino glycocide antibiotics derivatives - Google Patents

Abstract

The method of preparing aminoglycocide antibiotics comprises: (a) dissolving 1.46g of 2-hydroxyimino-2-phenylacetonitril in 20ml of tetrahydropyran at room temp; (b) adding 1.4ml of triethylamine to the mixture; (c) mixing 1.86g of diethylchlorophosphate with the above mixture for 4 hrs; (d) filtering the mixture and concentrating the resulting solution.

Description

Method for preparing aminoglycoside antibiotic derivatives

The present invention relates to an improved method for preparing an aminoglycoside antibiotic represented by the following structural formula (I), which is a known antibiotic.

It is an object of the present invention to acylate only one amino group in one of these amino groups in the molecule of kanamycin A having four active amiro groups. In the preparation of the compound of formula (I), there are many problems in selectively preparing the target compound because there is no significant difference in activity of each amino group. Therefore, various improvement methods have been invented to solve this problem, and the present invention is one of such improvement inventions.

The present invention provides a method of selectively acylating only 1-amino groups by reacting and reacting reactive silyl derivatives with kanamycin A, which protects 3,6'-amino groups, wherein the method of activating the acylating agent used herein It is the summary of the invention.

Acylating agents conventionally used include N-hydroxy-5-norbornene-2, as described in US Patent No. 3,781,268, Japanese Patent Application Publication No. 49-102,644, US Patent No. 4,424,343, US Patent No. 4,347,354, and the like. Activated and used as 3-dicarboxyimide, N-hydroxy succinimide or N-hydroxy phthalimide.

However, since the active derivatives of such acylating agents are expensive or difficult to obtain, there are many problems in practical use in the industry, and the overall yield of the acylating agents is only about 1%. come.

The present invention is to enable the acylating agent using 2-phenylacetonitrile-2-aminoyl-diethyl phosphate, which is very inexpensive and easily prepared, and to enable the activated acylating agent as a reactant. It also has advantages such as high yield of the target compound. If this is expressed in detail step by step as follows.

end. 1) The following structural formula (II) 'compound is prepared by a known method, for example, US Pat. No. 4,136,254 or Korean Patent No. 16,106.

2) Polysilylated Structural Formula (II) compounds are prepared by silylating the compounds of Structural Formula (II) 'by US Pat. No. 4,347,354 or Korean Patent No. 16,107.

R is a trialkyl silyl group, such as hydrogen or a trimethylsilyl group, etc. here.

I. 1) 2-hydroxyimino-2-phenylacetonitrile is reacted with disubstituted chlorophosphate to prepare 2-phenylacetonitrile-2-aminoyl-disubstituted phosphate represented by the following structural formula (III) '.

Z represents alkyloxy, phenoxy, benzyloxy, aryloxy and the like.

2) A compound of formula (III) is prepared by reacting a compound of formula (III) 'prepared by L-4-substituted amino-2-hydroxybutyric acid (formula (III)' ').

Here, B is an aryloxycarbonyl group such as benzooxycarbonyl and P-nitro benzyloxycarbonyl, an alkoxycarbonyl group such as butoxycarbonyl group, and a halo alkoxycarbonyl group such as trichloroethoxycarbonyl. n is an integer of 0-3.

All. The polysilylated compound (II) and the structural formula (III) are subjected to a direct reaction to prepare a compound of the following formula (IV), and then a protecting group is removed by a method described in known methods such as US Patent No. 3,781,268. (I) A compound is prepared.

R is a trialkylsilyl group, such as hydrogen or a trimethylsilyl group, etc. here. Compound (III) 'is a novel compound, and its intermediate, 2-hydroxyimino-2-phenyl-acetonitrile, is a known substance. Bulletin of Chemical society of Japan Vol. 50 (3) 718-721 (1977)) and organic synthesis (Org. Syn (II) Vol (II) 363), and sodium nitrite in an aqueous methanol solution in an alkaline alcoholic solvent, By absorbing and treating the resulting methyl-nitrite, the pale yellow oxime can be easily produced in high yield of 85% or more. When this is reacted with dimethyl crorophosphate in the presence of a base, a quantitative target (III) 'compound can be obtained.Tetra hedron Letters Vo. 21.1467-1468 (1980), Journal of Chemical society , chem.chmm 1029-1030P (1980) can be used.

The solvent can be reacted well in all of dimethyl formamide, acetone acetonitrile, tetrahydrofuran, methylene fluoride and the like, and the reaction temperature is about 0-100 ° C, preferably at room temperature. In addition, the (III) 'compound may be used directly in the next reaction without being separated. In preparing the compound of formula (III) ', free acid adsorption reagents include alkyl secondary base such as diethylamine, alkyl tertiary base such as triethylamine, or primary organic base such as n-butylamine. It is also possible to use, preferably triethylamine, and as the organic activating agent, an organic halogen such as dialkoxyhalophosphate such as diethyl chlorophosphate, other aryloxy, phenoxy, etc. may be used. Polophosphate is preferred.

Furthermore, as an active ester agent of (III) ', N-hydroxy succinimide, N-hydroxy-5- norbornene-2,3- dicarboxy-imide, N-hydrociphthalimide, N-hydroxy Benzotriazole etc. are also possible, but compound (III) 'is preferable.

The compound of formula IV can be prepared by a conventional method using compound III, and possible solvents are tetrahydrofuran, acetone, dimethyl sulfoxide, and dicyclohexylcarbodiimide at room temperature. hexyl-carbodiimide).

In preparing the compound of the above formula (IV), the compound of the formula (III) may be used, or the compound (III) '' and (III) 'may be directly acted on the compound (II) to shorten and simplify the process. Both are preferable, that is, the polysilylated structural formula (II) compound is a substance that has increased solubility and lipophilicity as described in the prior patent example, and can be easily applied to the operation we will perform. The compound of formula (III) can be acylated under mild conditions in the presence of a general organic reaction solvent, i.e., tetrahydrofuran, acetone, acetonitrile, dimethylformamide, and the like. Tetrahydrofuran is preferable as the solvent. The misfire reaction is possible from -10 ° C. to 100 ° C., but it is preferable at ambient temperature and the time is more than 2 hours. The exhalation is removed to obtain the compound of formula (I), ie, the silyl group can be removed near pH 2-3 after the addition of water, and the benzyloxycarbonyl group, an amino protecting group, is easily removed under hydrogen pressure at room temperature under a palladium carbon catalyst. If the protecting group is removed, column separation with CG 50 (NH ₄ ⁺ ) produces a pharmaceutically effective formula (I) compound and salts thereof.

It will be described in detail in the following examples.

Example 1

Preparation of 2-phenylacetonitrile-2-iminoyl-diethyl phosphate

1.46 grams of 2-hydroxyimino-2-phenylacetonitrile was added to 20 millimeters of tetrahydrofuran, dissolved at room temperature, and 1.4 milliliters of triethylamine were added. Then, 1.86 grams of diethylchlorophosphate was slowly added dropwise under ice-cooling, followed by vigorous stirring. The reaction was continued at room temperature for 4 hours, and then the crystals formed by cooling were filtered off. The filtrate is concentrated to dryness to give 2.88 grams of the desired product in the form of an oil. It is no longer purified and can be used for the next reaction.

(1) TLC: R / F 0.75 (benzine: tetrahydrofuran: formic acid = 15: 5: 2)

(2) IRνmax cm ^-1 : 2980, 1440, 1370, 1280, 1160, 950

(3) NMR: 1.4 (6H.m.CH ₃ ), 4.3 (4H.m.CH ₂ ), 7.5-7.9 (5H.m.benzyl)

Example 2

Preparation of L-4-benzyloxycarbonylamino-2-hydroxy-butyric acid-2-phenylacetonitrile-2-iminoester

1) After dissolving 1.226 grams of L-4-benzyloxycarbonylamino-2-hydroxy-butyric acid in 15 milliliters of tetrahydrofuran, 0.7 milliliters of triethylamine is added and stirred. Subsequently, 1.48 grams of 2-phenylaceto-nitrile-2-iminoyl-diethylphosphate dried in Example 1 at room temperature was dissolved in 5 milliliters of tetrahydrofuran, and then added to a reaction solution. After stirring for 6 hours at room temperature, the solvent was distilled off, dissolved in ethyl acetate, washed with brine, and then the organic layer was dehydrated with anhydrous forget-mean and crystallized with hexane to obtain 1.68 grams of the target substance.

(1) TLC: R / F 0.8

(2) MP: 92-96 ℃

(3) IRνmaxcm ^-1 : 3300, 1760, 1680, 1530, 1270

(4) NMR 60 MHZ: 2.1 (2H.m); 5.0 (2H.s), 3.4 (2H.m); 7.3 (10H.m), 4.5 (1H.m)

2) Dissolve 2.53 grams of L-4-benzyloxycarbonylamino-2-hydroxy butyric acid in 20 milliliters of tetrahydrofuran at room temperature. After adding 1.46 grams of 2-hydroxyamino-2-phenyl acetonitrile and dissolving, a solution obtained by dissolving 2.06 grams of dicyclohexyldiimide in 10 milliliters of tetrahydrofuran at room temperature is added dropwise from the dropping tank for 1 hour. The reaction is terminated by further stirring for 4 hours. The cake produced by cooling for a while is filtered off, and the filtrate is concentrated and hardened to crystallize in hexane to obtain 3.51 grams (92%) of white crystal powder. The analysis value is the same as in method 1).

Thin layer chromatography: GF254, R / F value: 0.8, benzene (15): tetrahydrofuran (5): formic acid (2)

Example 3

Preparation of 1-N- [L-(-)-4-amino-2-hydroxybutyryl] -ganamycin A

1.505 grams of 3,6'-N-dibenzyloxycarbonyl kanamycin A is suspended in 20 milliliters of acetonitrile, and then warmed to reflux. 2.25 milliliters of hexamethyldisilazane was added dropwise to the distillate, followed by heating under reflux for 7 hours, and removing the solvent to give 2.6 grams of white crystalline powder. After dissolving this silicide in 15 milliliters of acetone, 0.92 grams of the active ester prepared in Example 1 was dissolved in 8 milliliters of acetone at room temperature. At a temperature, 0.92 grams of the active ester prepared in Example 1 was dissolved in 8 milliliters of acetone. The reaction is terminated by stirring at room temperature overnight. Then, 15 ml of water was added, the reaction solution was adjusted to ph 2.0, stirred for about 30 minutes, and 20 ml of methanol was added to the pale yellow residue obtained by concentrating and removing the organic solvent. 10% pd / c 0.4 grams were added and hydrocracked under hydrogen pressure of 45 PSI for 4 hours at room temperature. The pd / c was filtered off, the filtrate was concentrated under reduced pressure to remove the organic solvent, the solution was neutralized, and adsorbed onto 120 ml of weakly prepared acidic cation exchange resin CG 50 (NH ₄ ⁺ ). Subsequently, the mixture was fractionated while eluting with water and 0,3N ammonia 0.5N ammonia water sequentially, followed by thin layer chromatography GF 254 (tetrahydrofuran (1): methanol (1): water (1): concentrated ammonia water (1)). R / F value: When only 0.16 is taken and concentrated under reduced pressure, 0.77.gram (64%) of white crystal powders which are the target objects are obtained.

(1) [α] _υ ²⁴ : + 98 ° (c = 1.0 H ₂ O)

(2) MP: 197-202 ° C (decomposition)

cm^-1: 3350, 2930, 1640, 1580, 1350(3) IRν

cm ^-1 : 3350, 2930, 1640, 1580, 1350

(4) NMR (δ ppm): 1.1-2.3 (4H.m), 2.6-3.9 (19H.m), 4.1 (1H.d), 5.21 (1H.d)

(5) Thin layer chromatography (GF 254 silica gel)

end. Developing solvent: (1) Methanol: Tetrahydrofuran: Water: Concentrated ammonia = 1: 1: 1: 1: 1 1 (2) Acetone: Pyridine: Glacial acetic acid: Water = 7: 3: 3: 35

I. Detection: 0.3% ninhydrin solution

All. R / F value

(6) Activity: 955 μg / mg (cylindrical plate method)

Example 4

Preparation of 1-N- [L (-)-4-amino-2-hydroxy-butyryl] -ganamycin A

1.505 grams of 3,6'-N-dibenzyloxycarbonyl kanamycin A is suspended in 20 milliliters of acetonitrile, and then warmed to reflux. 2.25 milliliters of hexamethyl disilazane was added dropwise while refluxing. Subsequently, after refluxing and cooling for 7 hours, the solvent was concentrated under reduced pressure to remove 2.6 grams of white crystalline powder. After dissolving the whole polysilicide in 20 milliliters of tetrahydrofuran, 0.608 grams of L-4-benzyloxycarbonylamino-2-hydroxybutyl acid and 0.243 grams of triethylamine are dissolved at room temperature. Subsequently, after dissolving 0.711 grams of 2-phenylacetonitrile-2-iminoyl-diethylphosphate prepared in Example 1 in 8 milliliters of tetrahydrofuran, the solution was added dropwise, followed by stirring at room temperature for 8 hours to terminate the reaction. Subsequently, 15 milliliters of water was added followed by treatment in the same manner as in Example 3 to obtain 0.685 grams (57%, 1/4 H ₂ CO ₃ base) of white crystalline powder as a target compound.

(1) [α] _D ^{23 °} : + 99 ° (c = 1.0 H ₂ O)

(2) Thin layer chromatography (GF 254 silica gel): R / F value = 0.16 (developing solvent of Example 3)

(3) Activity: 960 µg / mg (cylindrical plate method)

Example 5

Preparation of 1-N- [L-(-)-4-amino-2-hydroxybutyryl] -ganamycin A

Suspend 1.505 grams of 3,6'-N-dibenzyloxycarbonyl kanamycin A in 20 milliliters of acetonitrile, add 2.25 milliliters of hexamethyldisilazane, heat, complete and cool. After heating under reflux and stirring for 7 hours under the same conditions, the organic solvent was concentrated under reduced pressure and then dried to yield 2.6 grams of white crystals. Separately, 10 milliliters of acetone was added to dissolve 0.608 grams of L-4-benzyloxycarbonylamino-2-hydroxybutyryl acid and 0.243 grams of triethylamine. Next, 0.71 grams of 2-phenylacetonitrile-2-iminoyl-diethylphosphate prepared in Example 1 was dissolved in 8 milliliters of acetone, and then added to the solution at room temperature. After stirring for 5 hours at the same temperature, the reaction solution is used without further purification. The total amount of white crystals prepared above is dissolved in 15 ml of acetone, and 0.31 ml of diethylamine is added, followed by stirring at room temperature. Here, the active ester of oxime prepared separately is added at a time and reacted overnight at the same temperature. Subsequently, 15 milliliters of water was added followed by treatment in the same manner as in Example 3 to give 0.818 grams of white crystals as the target compound.

(1) [α] _υ ²³ : -99 ² (c = 1.0 H ₂ O)

(2) TLC (GF254) R F value: 0.16 (developing solvent of Example 3)

(3) Elemental analysis: as C ₁₂ H ₄₃ N ₅ O ₁₃ 1/4 H ₂ CO ₃

Experimental value: C; 44.01%, H; 7.31%, N; 11.55%

Theoretic value: C; 44.66%, H; 7.29%, N; 11.65%

Example 6

Preparation of 1-N- (L-(-)-4-amino-2-hydroxy butyryl) -ganamycin A-sulfate.

After dissolving 1 gram of 1-N- [L-(-)-4-amino-2-hydroxybutyryl] kanamycin A prepared in Example 4 in 5 milliliters of distilled water, the mixture was diluted with dilute sulfuric acid while gradually stirring at room temperature. Adjust to 6.6. After stirring for a while, 0.3 grams of activated carbon was added to the reaction mixture, decolorized at room temperature, filtered, and sterile filtered. The lyophilized solution obtained 1.2 grams of white crystals.

(1) [α] _ν ²² : + 76 ° (c = 1.0 H ₂ O)

(2) MP: 222-226 ° C (decomposition)

(3) ph: 6.0 (1% H ₂ O)

(4) Titer: 705 µg / mg

(5) Loss on drying: 1.5%

cm^-1: 3350, 2925, 1640, 1580, 1350(6) IRν

cm ^-1 : 3350, 2925, 1640, 1580, 1350

Claims (1)

It is characterized by a polysilylated compound of the following structural formula (II) to the acylating agent represented by the following structural formula (III) to produce the following structural formula (IV) compound, and hydrogenated to the compound represented by the following structural formula (I) A method for preparing aminoglycoside antibiotic derivatives and their esters or pharmaceutically effective nontoxic acid addition salts.

Here, B is an aryloxycarbonyl group such as benzooxycarbonyl and p-nitrobenzyloxycarbonyl, an alkoxycarbonyl group such as butoxycarbonyl group, and a haloalkoxycarbonyl group such as trichloroethoxycarbonyl. R is a trialkylsilyl group such as hydrogen or trimethylsilyl group and the like. n is an integer of 0-3.

R is a trialkylsilyl group, such as hydrogen or a trimethylsilyl group, etc. here.