JPS6044316B2 - 3″-deoxydihydrostreptomycin and its manufacturing method - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a new substance, 3''-deoxydihydrostreptomycin, and a method for producing the same.

Streptomycin is a prominent antibiotic discovered by Waksmann and is widely used in medicine, along with its aldehyde group reduced form, dihydrostreptomycin.

However, as they have been widely used, bacteria resistant to them have emerged, and this fact has significantly reduced the efficacy of streptomycin and dihydrostreptomycin. However, regarding this historical situation, there is a review by Hamao Umezawa, one of the inventors of the present invention and the first discoverer of the resistance mechanism described below. , In Carbohydrate Chemistry &
Biochemistry, Vol. 30, p. 183, Academic Press 1974jV-). In the case of streptomycins, the hydroxyl group at the 3″ position is adenylylated by streptomycin and adenylyl transferase-resistant bacteria, resulting in the formation of 3″-0-adenylylstreptomycin, which inactivates its drug efficacy. is [in the formula, Y
is a benzyloxycarbonyl group as a protecting group, Z is an acetyl group as a protecting group, and A is a cyclohexylidene group] is condensed, and the streptidine protected derivative represented by the following formula (■) [in the formula, R ．． A condensation product represented by R'', Y. A method for producing 3''-deoxydihydrostreptomycin and its acid addition salt represented by the following formula (1), which comprises reacting with a common acid to form an acid addition salt.

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a new substance, 3''-deoxydihydrostreptomycin, and a method for producing the same.

Streptomycin is a prominent antibiotic discovered by Waksmann and is widely used in medicine, along with its aldehyde group reduced form, dihydrostreptomycin.

However, as they have been widely used, bacteria resistant to them have emerged, and this fact has significantly reduced the efficacy of streptomycin and dihydrostreptomycin. It commonly occurs against mycins and other pharmaceuticals, but this historical situation can be found in a review by Hamao Umezawa, one of the inventors of the present invention and the first discoverer of the resistance mechanism described below. Advances●In●Carbohydrate Chemistry●and・
Biochemistry, Vol. 30, p. 183, Academic Press 1974jV-. ) in detail. In the case of streptomycins, the hydroxyl group at the 3'' position is adenylylated by streptomycin-resistant bacteria with adenylyl transferase, resulting in the formation of 31-0-adenylylstreptomycin, which inactivates its drug efficacy. (Umezawa et al., Journal of Antibiotics, 2)
Volume 1, page 81, 196 details). Therefore, the present inventors
By removing the hydroxyl group at the `` position, we have begun research to eliminate the possibility of inactivation due to adenylylation and obtain a streptomycin derivative that is effective against the streptomycin-resistant bacteria. We have succeeded in synthesizing dihydrostreptomycin and found that this substance is effective against resistant bacteria.We have previously found that this substance is effective against resistant bacteria. The present inventors have reported that removing the hydroxyl group that is phosphorylated or adenylylated by resistant bacteria can convert it into a derivative that is effective against resistant bacteria. It is also known that the antibacterial activity of antibiotics itself may be lost.However, the 3''-deoxydihydrostreptomycin of the present invention does not cause this decrease in antibacterial activity and is effective against resistant bacteria compared to streptomycin. This was the first demonstration of matter. The first gist of the present invention is 3''-deoxydihydrostreptomycin and its acid addition salt represented by the following formula.

The 3''''-deoxydihydrostreptomycin (free base) of the present invention is a colorless solid material and is relatively unstable.

This trihydrochloride is a stable colorless solid substance, and its specific optical rotation [α] is 193 (C1, water). 3
Table 1 shows the antibacterial activity of ``''-deoxydihydrostreptomycin in comparison with that of dihydrostreptomycin.

The gist of the second invention is as follows:
3''-deoxydihydrostreptobiosaminyl represented by [wherein X is a halogen atom, R is a known monovalent amino protecting group, and R'' is a known monovalent hydroxyl protecting group]
A streptidine protected derivative represented by the following formula (■) [wherein Y represents a benzyloxycarbonyl group as a protecting group, Z represents an acetyl group as a protective group, and A represents a cyclohexylidene group] is condensed with a halide derivative. Then, the following formula (
■)ノ [In the formula, R. ．． A condensation product represented by R'', Y.. A method for producing 3''-deoxydihydrostreptomycin of formula (1) and its acid addition salt, which is characterized by reacting with a common acid to form an acid addition salt by a conventional method.

To describe in detail the method of the present invention, a 3''-deoxydihydrostreptobiosaminyl halide derivative represented by the formula (■), which has already been deoxylated at the ``3'' position, is used as a starting material.

The compound of formula (■) can be prepared as described in Reference Examples 1 to 13 below.

When acetic anhydride is used as in Reference Example 6, a product in which R is an acetyl group is obtained, and in Reference Example 6, when an acylic anhydride is generally used in place of acetic anhydride, a product in which R is the corresponding acyl group is obtained. It will be done. When an alkyl chloroformate or a corresponding alkyl chloroformate is used in place of anhydrous sodium acetic acid monoacetate in Reference Example 6, a product in which R is the corresponding alkyloxycarbonyl or aralkyloxycarbonyl group is obtained. Furthermore, when acetic anhydride is used as in Reference Example 10, a product in which R'' is an acetyl group can be obtained, and Reference Example 1
When an acylating agent that generally contains an acyl group R'' is used in place of acetic anhydride of 0, a compound (■) containing the corresponding acyl group as R'' is obtained.

Known substance di-N-acetyl-di-N-benzyloxycarbonyl-5 represented by formula (■) for compound (■)
・6-0-Cyclohexyristreptidine (racemate) (see Umezawa et al., Pretans of the Chemical Society of Japan, Kitamaki, p. 560, 197 tsubo) was condensed using a conventional method for oligosaccharide synthesis. The product of formula (■) is obtained as a condensate of both. After purifying this condensate (■), its protective group Y. Z. ．． R and R″ and 3・C
When the CO group bonded to the hydroxyl group is removed, the desired 3''''-deoxydihydrostreptomycin of formula (1) is produced. A chlorine atom or a bromine atom is usually used as X in formula (■). Also, R. ．． R'' is a lower alkanoyl group such as an acetyl group or a pteroyl group, an acyl group such as an aroyl group such as a benzoyl group, or a lower alkoxycarbonyl group such as an ethoxycarbonyl group or an aralkyloxycarbonyl group such as a benzyloxycarbonyl group. etc. The condensation reaction between compounds (■) and (■) is preferably carried out in a solvent, and the solvent does not wait for active hydrogen, and the condensation reaction between compounds (■) and (■) is preferably carried out in a solvent.
Any solvent that at least partially dissolves (2) or (2) can be used. Examples include dioxane, chloroform, dichloromethane, and benzene. Pyridine, mercury cyanide, silver carbonate, etc. are used as the condensation catalyst. Reaction temperature: O'C~80'C1 Reaction time: 1 hour~7
The day is appropriate. Since the reactant (■) is a racemate, products condensed at the 4-position (target product) and 6-position of the streptidine moiety are produced, but these can be easily separated by column chromatography. The condensation product (■) is then removed from the protective group, and its O-acyl group (R″)
, N-acyl group (Z) bonded to the guanidine group and 3.
The carbonyl group bonded to the C-position hydroxyl group is easily removed by the action of ammonia, alkali hydroxide, calcium hydroxide, barium hydroxide, etc. In addition, a benzyloxycarbonyl group (
Y) is removed by alkaline decomposition, which is more severe than the above, or by catalytic hydrolytic decomposition such as palladium black. Further, the cyclohexylidene group (A) is removed by applying a dilute acid such as aqueous acetic acid or dilute acetic acid. Further, the N-acetyl group (R) of the dihydrostreptobiosamine moiety is removed by the action of a basic reagent such as barium hydroxide or sodium amide. The crude 3''-deoxydihydrostreptomycin thus obtained is purified by resin treatment, etc., and if desired, ordinary acids such as hydrochloric acid, phosphoric acid, sulfuric acid, and citric acid can be used in a conventional manner, making it pharmaceutically acceptable. It can also be used in the form of a suitable acid addition salt.Example 1 (a) 4-0-(N-acetyl-4''.6''-di0-
Production of acetyl-3●C-0-carbonyl-3''-deoxydihydrostreptobiosaminyl)-di-N-acetyl-di-N-benzyloxycarbonyl-5,6-0-cyclohexylidene streptomycin.

N-acetyl 4″・6″-G0-acetyl 3・C
121 mg of -0-carbonyl-3''-deoxydihydrostreptobiosamine (see Reference Example 13 below) was dissolved in 3 mt of thionyl chloride and left in the dark at room temperature for a specific time.

The reaction solution was concentrated under reduced pressure at room temperature to quantitatively obtain the corresponding 1-chloride derivative as a glassy substance. Di-N-acetyl-di-N-benzyloxycarbonyl-5●6 (and 4,5)-0-cyclohexylidene streptidine (racemic form, 154 mg) was added to this 1-chloride, and 1.9 mL of anhydrous dichloromethane was added. 1 was added and dissolved, and further 110 m9 of anhydrous calcium sulfate, 110 m9 Molecular Sieve 4A, and 140 m9 mercuric cyanide were added, and the mixture was shaken at room temperature for 4 days to carry out a condensation reaction. 19 ml of chloroform was added to the reaction solution, and after filtering the solution, the solution (1) was washed with a saturated aqueous solution of sodium bicarbonate and then with water. After drying, the solution was concentrated to obtain a glassy substance. The product mixture was separated by silica gel column chromatography using benzene-methyl ethyl ketone (2:1) as the developing system. The fraction containing the first substance in the elution order was collected and concentrated to yield 30 mg of the title substance as a glassy solid. (b) 3″″-deoxydihydrostreptomycin, where Ac = acetyl group 18 m9 of the substance obtained in (a) above was mixed with 0.6 ml of ammonia-saturated methanol.
The acetyl group bonded to the guanidine group, the 0-acetyl group, and the carbonyl group bonded to 3◆() were removed by dissolving it in and leaving it at room temperature for 5 hours.

The glassy substance obtained by concentration was dissolved in dioxane, and hydrogenolyzed under a hydrogen pressure of 3 atmospheres using palladium black as a catalyst to remove the benzyloxycarbonyl group. Reaction liquid? After filtering and concentrating, the resulting glassy solid was dissolved in 50% acetic acid and heated to 55°C for 3 hours to remove the cyclohexylidene group. Next, the reactivity was concentrated under reduced pressure, and the obtained solid was dissolved in a 5.5% barium hydroxide solution at 60°C. 2.
The reaction was carried out for a period of time to remove the N-acetyl group.

After introducing carbon dioxide gas, the reaction solution was filtered, the bottom liquid was concentrated, the obtained glassy solid was dissolved in water, and the solution was placed in a strong basic resin (Dowex 1×2 (0H type)) column. , extract a fraction containing the target substance by diluting with water, concentrate this fraction, neutralize it with hydrochloric acid, and further concentrate.
3.1 mg of 3″-deoxydihydrostreptomycin was obtained as trihydrochloride. [α]D-93 salt (C1,
water). Reference Example 1 Production of pentyl N-benzyloxycarbonyl dihydrostreptobiosaminide 6.81 y of benzyl dihydrostreptobiosaminide hydrochloride was dissolved in 90 ml of water, and 90 ml of acetyl and 3.2 q of sodium carbonate were added. , 4.6 ml of benzyloxycarbonyl chloride under ice cooling
added.

The precipitated solid was recrystallized from chloroform-ether to obtain the title compound as granular crystals (5.56 g). [α] Basa 4
-1140. (C1, chloroform). Reference example 2 Pentyl N-benzyloxycarbonyl 3.C; 4
Production of ``6''-di-0-isopropylidene dihydrostreptobiosaminide Substance obtained in Reference Example 1 6.72y
and 960 ml of p-toluenesulfonic acid were dissolved in 120 ml of anhydrous dimethylformamide, 19 ml of 2,2-dimethoxypropane was added, and the mixture was reacted at 50°C for 3 hours.

Pour the reaction solution into a mixture of 5% sodium hydrogen carbonate solution and chloroform, remove the chloroform layer, wash with water,
After drying, the solvent was distilled off to obtain the title compound as an oily substance 7.3y. [α] Basa 4-1173 (C1, chloroform). Reference example 3 Pentyl N-benzyloxycarbonyl-3.C; 4″
・Production of 6″-di0-isopropylidene-3″-0-mesyldihydrostreptobiosaminide Substance 7.28y obtained in Reference Example 2 was dissolved in pyridine and treated with methanesulfonyl chloride by a conventional method, The title compound was obtained as a glassy material 8.14y.

Reference Example 4 Pentyl 2-0-(2.3-N.0-carbonyl-2-deoxy-4●6-0-isopropylidene-2
-methylamino-α-allopyranosyl)-3●C-0
- Substance 8.14y obtained in Reference Example 3 for the production of isopropylidene dihydrostreptoside and sodium acetate, 3
8.14q of aqueous salt was dissolved in 160ml of 2-methoxyethanol and heated under reflux for B hours.

The reaction solution was concentrated under reduced pressure, the resulting solid was dissolved in chloroform, and this solution was washed with water. After drying the solution, the solvent was distilled off, and the obtained solid was recrystallized from ethanol to obtain the title compound. Yield 5.17ymp. 171-172℃, [
α〕? -171 amount (C1, chloroform). Reference Example 5 Production of pentyl 2-0-(2-deoxy-4●6-0-isopropylidene-2-methylamino-α-L-allopyranosyl)-3●C-0-isopropylidene dihydrostreptoside Obtained in Reference Example 4 1.04y of material was refluxed in 1N ethanolic potassium hydroxide for 4 hours to give the title compound as 640m9 of decarbonylated product.

Mp. llO~. 1116C1 [α] Kun-131 go (C
1, chloroform). Reference Example 6 Pentyl 2-0-(N-
Acetyl-2-deoxy-4,6-0-isopropylidene-2-methylamino-alpha-allopyranosyl)-3●
Production of C-0-isopropylidene dihydrostreptoside 652 mg of the substance obtained in Reference Example 5 was dissolved in methanol and treated with anhydrous sodium acetic acid monoacetate in a conventional manner to obtain the above-titled N-acetyl compound as an oily substance. .

Yield 665m9. [α] Basa 3-137 (C1, chloroform). Reference Example 7 Production of pentyl N-acetyl-3″-chloro-3″-deoxy-3·C;4″·6″-0-isopropylidene dihydrostreptobiosaminide Substance 4 obtained in Reference Example 6
87Tng was dissolved in 5.3TrL1 of anhydrous dichloromethane, 0.9mL of pyridine was added, the solution was cooled to -5°C, 0.35mt of sulfuryl chloride was added, and the reaction solution was cooled to -5°C.
The mixture was left at 5° C. for 1 hour and then at 5° C. for 2 hours.

The reaction solution was poured into a mixture of saturated aqueous sodium hydrogen carbonate and chloroform, stirred, and the chloroform layer was removed.
After drying, the solvent was distilled off. The obtained oily substance was purified by silica gel column chromatography using benzene-methyl ethyl ketone as the developing system. Recrystallization from ether gave the title compound as granular crystals (284 mg, 56.4%). Mp. l7O~177C1 [α] Bas4-12
9 items (C1, chloroform). Reference example 8 Pentyl N-acetyl-3″-deoxy-3・C;4″
●Production of 6″-di0 isopropylidene dihydrostreptobiosaminide 1.01y of the substance obtained in Reference Example 7 was dissolved in 21Tn1 of anhydrous toluene, and the
-butyltin hydride 1.0mt1 then α・α2
- 10 mg of azobisisobutyronitrile was added, and the mixture was heated and stirred at 80°C for 2 hours.

The oily substance obtained after concentrating the reaction solution was purified by silica gel column chromatography using benzene-methyl ethyl ketone 4:1 as the developing system to obtain 935 mg of the title compound as a glassy substance. [α]? -1000(C1,
chloroform). Reference Example 9 Pentyl N-acetyl 3″
-Manufacture of deoxy-3.C-0-isopropylidene dihydrostreptobiosaminide Substance 93 obtained in Reference Example 8
2m9 was mixed with methanolic 25% acetic acid and heated under reflux to obtain 821m9 of the title compound as a glassy substance.

[α]? -1611 (C1, chloroform). Reference example 1
Production of 0-pentyl N-acetyl-4'', 6''-di0-acetyl-3''-deoxy-3, C-0-isopropylidene dihydrostreptobiosaminide The material obtained in Reference Example 9 was dissolved in acetic anhydride in pyridine by a conventional method. The title compound was obtained in 95% yield.

[α]? -1400 (C1, chloroform). Reference example 1
Production of 1-pentyl N-acetyl-4''●6''-di-0-acetyl-3''-deoxydihydrostreptobiosaminide 1.73y of the substance obtained in Reference Example 10 was dissolved in 70% acetic acid 4
After dissolving in 3Tn1 and heating at 80'C for 3 hours, the reaction solution was concentrated, and the resulting oily substance was dissolved in benzene-ethanol (9:
1) was purified by silica gel column chromatography as a developing system, and ether was added to the obtained oily substance.
Crystallization gave the title compound.

Yield: 811 mg, Mp. l57~159 gain C1 [α]?
5-169 chloroform). Reference Example 12 Production of pentyl N-acetyl-4''/6''-di-0-acetyl-3/C-0-carbonyl-3'-deoxydihydrostreptobiosaminide Substance 2 obtained in Reference Example 11
51m9 was dissolved in 7.5ml of pyridine, 115mg of p-nitrophenoxycarbonyl chloride was added, and further 0.16ml of triethylamine was added.

After 1 hour, 21 hours, and 2 hours, 1 g of p-nitrophenoxycarbonyl chloride and 0.15 ml of triethylamine were added, respectively.

After 4 hours, the reaction solution was diluted with chloroform, washed with a saturated aqueous solution of sodium bicarbonate and then with water, dried, and concentrated under reduced pressure.

The obtained oily substance was purified by silica gel column chromatography using benzene-methyl ethyl ketone (3:1) as a developing system to obtain the title compound as a glassy substance 225m9. Reference example 13 N-acetyl 4″・6″-di0-acetyl 3・C
Production of -0-carbonyl-3''-deoxydihydrostreptobiosamine 103 mg of the substance obtained in Reference Example 12 was dissolved in 3.5 mt of dioxane, palladium black was added,
Reduction was carried out under atmospheric hydrogen pressure at room temperature.

Claims (1)

[Claims] Primary formula (I) ▲ Numerical formulas, chemical formulas, tables, etc. ▼ 3″-deoxydihydrostreptoycin and its acid addition salt represented by (I). Secondary general formula (II) ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (II) [In the formula, X represents a halogen atom, R represents a known monovalent amino-protecting group, and R' represents a known monovalent hydroxyl-protecting group] 3' - Deoxydihydrostreptobiosaminyl halide derivative has the following formula (III) ▲ Numerical formula, chemical formula, table, etc. ▼
(III) [In the formula, Y represents a benzyloxycarbonyl group as a protecting group, Z represents an acetyl group as a protective group, and A represents a cyclohexylidene group] is condensed to form the following formula: (IV) ▲Mathematical formula, chemical formula,
There are tables, etc. ▼ (IV) A condensation product represented by [in the formula, R, R', Y, and Z have the meanings described above] is prepared, and the above protecting group is removed from it by a known method to obtain 3'' - Deoxydihydrostreptomycin is produced and, if desired, is further reacted with a common acid to form an acid addition salt in a conventional manner. ) A method for producing 3''-deoxydihydrostreptomycin and its acid addition salt.