JPH0154357B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention is based on the general formula (In the formula, R ₁ and R ₂ are different and represent a hydrogen atom or a methyl group, R ₃ represents a hydrogen atom or a glycyl group,
The present invention relates to a novel amino glycoside represented by (the amino group may be protected). For example, a compound of formula (In the formula, R ₃ ' represents a hydrogen atom or an acyl group, and R ₁
and R ₂ have the above-mentioned meanings, and the amino group may be protected) is treated with an acid to cleave the methyl ether, and if R ₃ is a hydrogen atom, the group - It can be produced by acylating NH- _CH3 and/or removing the protecting group. Among the compounds of the formula, the compound in which R ₁ is a methyl group and R ₂ is a hydrogen atom is one of the KA-6606 substance group (KA -6606 and the same) (see the specification of JP-A-55-111497).
Compounds in which R ₁ is a hydrogen atom and R ₂ is a methyl group are:
It is one of the KA-7038 substance group (KA-7038, KA-7038 and KA-7038) produced as a metabolite of the KC-7038 strain belonging to the genus Streptomyces.
54-141701 and 55-162795). All of the above antibiotics () have excellent antibacterial activity, but the de-O-methyl compound (), which is obtained by cleaving the methyl ether group, shows even more excellent antibacterial activity and is useful as a medicine (Japanese Patent Application Laid-Open No. 55-2010) −
55198 and 53-95943). In producing the compound of the present invention, the compound of the formula or its protected compound is first treated with an acid in the presence or absence of a solvent. This causes cleavage of the methyl ether and elimination of the acyl group such as the glycyl group bonded to the methylamino group, producing a compound of the formula in which R ₃ is a hydrogen atom. Examples of acids include hydrobromic acid, hydrochloric acid, hydroiodic acid,
Mineral acids such as hydrofluoric acid, sulfuric acid, and phosphoric acid, strong acidic organic acids such as p-toluenesulfonic acid and trifluoromethanesulfonic acid, and Lewis acids such as boron trichloride and boron tribromide are used. When a Lewis acid is used, it is preferable to carry out the reaction under anhydrous conditions, and in other cases, it is preferable to carry out the reaction in an aqueous solution. For example, dichloromethane is used as the anhydrous solvent. The reaction takes place at room temperature to 200°C, usually for 1 to 30°C.
Finish in time. The product can be separated and purified by conventional column chromatography using, for example, a cation exchange resin. By acylating the methylamino group at the 4-position of the thus obtained compound of the formula in which R ₃ is a hydrogen atom, a compound of the formula in which R ₃ is an acyl group is obtained. The acyl group is preferably one derived from an amino acid such as a glycyl group, an alanyl group, or a valyl group. These amino groups may be substituted with, for example, a lower alkyl group, a carbamoyl group, a formyl group, or the like. When carrying out acylation, after protecting the three amino groups at the 1-position, 2'-position and 6'-position present in the compound (R ₃ = H), substituted carboxylic acid or It is preferable to allow the reactive derivative to act, and then, if necessary, remove other protecting groups for the amino group to liberate it. As the protecting group for the amino group, those used in ordinary peptide synthesis can be used. For example, substituted phenyl esters of carbonic acid monobenzyl esters, N
When active esters such as -oxysuccinimide ester and N-oxyphthalimide ester are used, only the amino groups at the 1-, 2'- and 6'-positions are protected with benzyloxycarbonyl groups. At this time, it is preferable to include a metal compound such as nickel acetate, cobalt acetate, copper acetate, or the like. Protective groups such as substituted benzyloxycarbonyl group and tertiary butoxycarbonyl group can also be used. If the methylamino group at position 4 is protected at the same time, for example, this product is treated with an alkali to form a cyclic carbamate between the methylamino group at position 4 and the hydroxyl group adjacent to it, and this is then hydrolyzed. By doing so, only the methylamino group at the 4-position can be released. To introduce an acyl group into the methylamino group of the thus obtained compound with the formula in which the amino groups at the 1-, 2'-, and 6'-positions are protected (R ₃ = H), ordinary peptide synthesis methods are applied. For example, the amino group can be acylated using a protected amino acid or other substituted carboxylic acid or a reactive acid derivative thereof. As reactive acid derivatives, acid halides,
Active esters such as phenyl ester, cyanomethyl ester, N-oxysuccinimide ester, N-oxyphthalimide ester, acid azides, acid anhydrides, mixed acid anhydrides and others used in peptide synthesis can be utilized. As the amino protecting group for the amino acid, the same ones as the above-mentioned protecting groups are used, and it is preferable to use the same ones. In order to optionally remove the protecting group for the amino group from the thus obtained compound having the formula in which the amino group is protected, it is preferable to use, for example, a catalytic reduction method. As the catalyst, for example, palladium, platinum, Raney nickel, rhodium, ruthenium, nickel, etc. are used. Furthermore, if desired, a compound in which R ₃ is a substituted alkyl group can also be produced by reducing the acyl group of the compound in which R ₃ is an acyl group obtained by the above method. The reduction reaction is preferably performed before the elimination reaction of the protecting group of the amino group, and a reduction method using a reducing agent such as lithium aluminum hydride, sodium borohydride, diborane, etc. can be used. Isolation and purification of the compound of the formula, which is the target substance, is carried out by conventional methods, but it is preferable to use column chromatography. As an adsorbent, for example,
CM-Sephadex, Amberlight IRC-50,
Amberlight IRC-84, Amberlight CG-
50, it is preferred to use a cation exchange resin such as carboxymethyl cellulose. The development can be carried out by a concentration gradient method or a concentration step method using an alkaline aqueous solution, such as an ammonia aqueous solution or an ammonium diate aqueous solution, as a developing solvent. By collecting the active fraction from the eluate and lyophilizing it, a pure product of the compound can be obtained. Although the target compound can also be obtained in the form of an acid addition salt through purification operations, a free base type compound can be converted into an acid addition salt by a conventional method. Examples of acids for this purpose include inorganic acids such as sulfuric acid, hydrochloric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, carbonic acid, and nitric acid, or acetic acid;
Organic acids such as fumaric acid, malic acid, citric acid, mandelic acid, and succinic acid are used. All of the compounds of the formula obtained by the method of the present invention exhibit excellent antibacterial activity and are useful as antibacterial substances in medicines, veterinary drugs, etc., and are also useful as starting materials for synthesizing various derivatives. As a typical example, 5-de-O-methyl-4
-N-glycyl-KA-6606 (compound A) and 5-de-O-methyl-KA-7038 (compound B)
The following table shows the antibacterial spectrum of

【table】

[Table] Example 1 (a) De-O-methyl-KA-6606: 350 mg of the free base of KA-6606 was dissolved in 3.5 ml of 56% hydriodic acid and heated at 60°C for 4 hours in a sealed tube. .
The reaction solution was concentrated to dryness under reduced pressure, and the residue was dissolved in water and neutralized with concentrated aqueous ammonia. This solution is CM−
It is adsorbed on a column packed with 30 ml of Cephadex C-25 (NH ₄ ⁺ type) and eluted with aqueous ammonia ranging from 0.25N to 0.35N. Fractions containing the target substance are collected and concentrated to dryness, resulting in de-O-methyl-KA-
6606265 mg is obtained. Elemental analysis value: C ₁₄ H ₃₀ N ₄ O ₄・H ₂ O Calculated value (%) 49.98 9.59 16.65 Actual value (%) 49.65 9.44 16.61 Specific optical rotation: [α] ²³ _D +87° (C1, H ₂ O) NMR value: δ _D2O ppm 1.54 (3H, d, J = 6.5Hz, C-C H ₃ ) 2.85 (3H, s, N-C H ₃ ) 5.56 (1H, d, J = 3.3Hz, Anomeric H) (b) 1,2',6'-Tris-N-benzyloxycarbonyl-4-N-benzyloxycarbonylglycyl-de-O-methyl-KA-6606: De-O-methyl-KA-6606163mg was dissolved in 5 ml of methanol, 273 mg of nickel acetate was added, and the mixture was stirred at room temperature for 30 minutes. Then, 454 mg of N-benzyloxycarbonyloxysuccinimide was added.
Stir at the same temperature for 2 hours. Add 2.5 ml of concentrated aqueous ammonia to the reaction mixture and stir for an additional 2 hours. The reaction solution was concentrated to dryness under reduced pressure, and the residue was dissolved in chloroform 20
ml of 3N-ammonia water and shake. Separate the chloroform layer and dilute with 3N aqueous ammonia.
After washing twice with water and drying, the solvent was distilled off. Dissolve the residue in 9 ml of dioxane and add 330 mg of 2,4-dinitrophenyl ester of N-benzyloxycarbonylglycine and 0.3 ml of triethylamine.
Add and heat at 60℃ for 1 hour. After adding 1 ml of concentrated aqueous ammonia to the reaction solution and allowing it to stand for 1 hour, the solvent was distilled off. Dissolve the residue in 20ml of chloroform,
After washing three times with 0.5N sodium hydroxide and twice with water, drying and distilling off the solvent. The residue was subjected to silica gel column chromatography and eluted with chloroform-methanol (50:1). When fractions containing the target substance are collected and concentrated, a colorless solid of 1,
2',6'-Tris-N-benzyloxycarbonyl-4-N-benzyloxycarbonylglycyl-
288 mg of De-O-methyl-KA-660 is obtained. Elemental analysis value: C ₄₈ H ₅₇ N ₅ O ₁₃ Calculated value (%) 63.22 6.30 7.68 Actual value (%) 63.43 6.21 7.44 Specific rotation: [α] ²³ _D +32° (C1, CHCl ₃ ) NMR Value: δ _CDCl3 ppm 2.92 (3H, s, N-C H ₃ ) 1.21 (3H, d, J=6Hz, CH-C H ₃ ) (c) De-O-methyl-4-N-glycyl-KA-
6606: Dissolve 288 mg of 1,2',6'-tris-N-benzyloxycarbonyl-4-N-benzyloxycarbonylglycyl-de-O-methyl-KA-6606 in 4 ml of acetic acid and add 50 mg of palladium black. Catalytic reduction at room temperature. Filter the reaction solution and add 400% water to the solution.
ml, neutralized with aqueous ammonia, and diluted with CM-
It was applied to a column of Sephadex C-25 (NH ₄ ⁺ type) and developed by the concentration gradient method between 0.05N and 0.35N aqueous ammonia. Fractions containing the target substance are collected and lyophilized to form a colorless solid, De-O-methyl-
105 mg of 4-N-glycyl-KA-6606 is obtained. Elemental analysis value: C ₁₆ H ₃₃ N ₅ O ₅・H ₂ O Calculated value (%) 48.84 8.97 17.80 Actual value (%) 48.55 8.83 17.48 Specific rotation: [α] ²³ _D +115° (C1, H ₂ O) NMR value: δ _D2O ppm 1.52 (3H, d, J = 6.5 Hz, C-C H ₃ ) 3.62 (3H, s, N-C H ₃ ) 5.40 (1H, d, J = 3 Hz, anomeric H) Example 2 (a) DeO-methyl-KA-6606: 100 mg of KA-6606 was dissolved in 5 ml of 48% hydrobromic acid and allowed to stand at 37°C for 10 days. The reaction solution was concentrated to dryness at below 37°C, the residue was dissolved in 50 ml of water, and after neutralization with aqueous ammonia, CM-Sephadex C-25
(NH ₄ ⁺ type) column and developed using a concentration gradient method with 0.05N and 0.5N aqueous ammonia. When the part containing the target substance is concentrated, de-O-methyl-
KA-660619mg is obtained. The properties of this product were all consistent with those of de-O-methyl-KA-6606 obtained in Example 1a. (b) 1,2',6'-Tris-N-benzyloxycarbonyl-4-N-benzyloxycarbonylglycyl-de-O-methyl-KA-6606: 19 mg of de-O-methyl-KA-660 in methanol 0.8 ml, add 90 mg of benzyl p-nitrophenyl carbonate, and stir overnight at room temperature.
Add 30% methylamine-ethanol solution to the reaction mixture.
After adding 0.1 ml and stirring for an additional hour, the reaction solution was concentrated to dryness, and the residue was dissolved in chloroform, washed with water, and dried. Dissolve this in 1 ml of dioxane,
Add 0.05 ml of triethylamine and 35 mg of N-hydroxysuccinimidyl-N-benzyloxycarbonylglycine and heat at 80°C for 5 hours. The reaction solution is concentrated to dryness, the residue is dissolved in chloroform to remove insoluble matter, the chloroform layer is washed with water and dried, and the solvent is distilled off. The residue was separated and purified by silica gel preparative chromatography (chloroform-methanol 15:1) to yield 1,2',6'-tris-N-benzyloxycarbonyl-4-N-benzyloxycarbonyl as a colorless solid. 623 mg of glycyl-de-O-methyl-KA-660 is obtained. The properties of this product were consistent with those of the compound obtained in Example 1b. (c) De-O-methyl-4-N-glycyl-KA-
6606: 1,2',6'-Tris-N-benzyloxycarbonyl-4-N-benzyloxycarbonylglycyl-de-O-methyl-KA-660623 mg in acetic acid
Dissolve in 0.5 ml, add 10 mg of palladium black, and perform catalytic reduction at room temperature. Filter the reaction solution and add 50ml of water.
After diluting with CM-
It was applied to a column of Sephadex C-25 (NH ₄ ⁺ type) and developed by the concentration gradient method between 0.05N and 0.4N aqueous ammonia. Fractions containing the target substance are collected and lyophilized to obtain 66067 mg of De-O-methyl-4-N-glycyl-KA-6 as a colorless solid. The properties of this product were consistent with those of the compound obtained in Example 1c. Example 3 De-O-methyl-KA-7038: Example 3 using 302 mg of the free base of KA-7038.
After reaction and purification in the same manner as in 1a, 220 mg of De-O-methyl-KA-7038 is obtained as a colorless powder. Elemental analysis value: C ₁₄ H ₃₀ N ₄ O ₄・H ₂ O Calculated value (%) 49.98 9.59 16.65 Actual value (%) 49.71 9.73 16.33 Specific optical rotation: [α] ²⁵ _D +40° (C0. 5, H ₂ O) NMR value: δ _D2O ppm 2.80 (3H, s, N-C H ₃ ) 2.84 (3H, s, N-C H ₃ ) 5.53 (1H, d, J = 3.3 Hz, anomeric H) Example 4 (a) Tetrakis-N-benzyloxycarbonyl-de-O-methyl-KA-7038: Using 190 mg of de-O-methyl-KA-7038,
When reacted and purified in the same manner as in Example 1b,
Colorless solid 1,2',6'-tris-N-benzyloxycarbonyl-4-N-benzyloxycarbonylglycyl-de-O-methyl-KA-7038,
That is, 350 mg of tetrakis-N-benzyloxycarbonyl-de-O-methyl-KA-7038 is obtained. Elemental analysis value: C ₄₈ H ₅₇ N ₅ O ₁₃ Calculated value (%) 3.22 6.30 7.68 Actual value (%) 63.01 6.49 7.42 Specific rotation: [α] ²⁵ _D +55° (C1, CHCl ₃ ) NMR Value: δ _CDCL3 ppm 2.92 (6H, s, 2×N-C H ₃ ) (b) DeO-methyl-KA-7038: Tetrakis-N-benzyloxycarbonyl-
When 320 mg of De-O-methyl-KA-7038 was treated in the same manner as in Example 2c, a colorless solid De-O-
112 mg of methyl-KA-7038 is obtained. Elemental analysis value: C ₁₆ H ₃₃ N ₅ O ₅・H ₂ O Calculated value (%) 48.84 8.97 17.80 Actual value (%) 48.54 8.69 17.98 Specific rotation: [α] ²⁵ _D +126° (C1, H ₂ O) NMR value: δ _D2O ppm 2.83 (3H, s, 6'-N-C H ₃ ) 3.63 (3H, s, 4-N-C H ₃ ) 5.42 (1H, d, J=3Hz, anomeric H)

Claims (1)

[Claims] 1. General formula (In the formula, R ₁ and R ₂ are different and represent a hydrogen atom or a methyl group, R ₃ represents a hydrogen atom or a glycyl group,
(The amino group may be protected) or its acid addition salt.