KR830001613B1 - The production of a selectively protected n-acylated derivative of an amino glycoside - Google Patents

Abstract

Prodn. of a selectively acylated N-protected deriv. of an aminoglycosidic antibiotic (I) comprises (a) reacting an acylating reagent having an acyl group to be introduced as the amine protecting group with an aminoglycosidic antibioticzinc cation complex to form a complex of Zn cations with the selectively N-acylated deriv. of the aminoglycosidic antibiotic having the initially non-complexed amino groups acylated; and (b) reacting the complex with a reagent which removes Zn cations from the complex to produce I. I consists of a 6-0 (3-amine- or 3-alkylamino-, 3-deoxyglycosyl) 2-deoxystreptamine and the selectively acylated N-protected deriv.

Description

Method for preparing aminoglycoside antibiotics selectively protecting amino groups

The present invention relates to a novel process for the preparation of protective derivatives of aminoglycoside antibiotics which selectively protects amino or alkylamino groups at specific positions of the aminoglycoside material with acyl groups. The present invention also relates to a method for selectively protecting amino or alkylamino groups at specific positions of aminoglycoside antibiotics.

The present invention mainly applies to the preparation of N-protective derivatives of aminoglycoside antibiotics of deoxystrepamines having 3 " -aminoglycosyl groups on hydroxyl groups at the 6-position. Aminoglycosides usable in the present invention Antibiotics are especially 6-C- (3 "-amino or 3" -alkylamino-3 "-deoxyglycosyl) -2-deoxystrep with 4-C- (6'-aminoclikyosil) substituents Aminoglycoside antibiotics composed of triamines, representative examples being kanamycin, gentamicin, sisomicin, netylmycin and verdamicin. The present invention also relates to the preparation of 1-N- (α-hydroxy-W-aminoalkanoyl) -amino-glycoside antibiotics known as useful semi-synthetic antimicrobials having activity against drug-resistant bacteria. Involves using new methods.

Conventional aminoglycoside antibiotics, such as kanamycin, are those that contain several relatively reactive amino and hydroxyl groups. Many kinds of semi-synthetic aminoglycoside antibiotics derived from parent aminoglycoside antibiotics have been prepared. In the semi-synthesis of such derivatives, it is preferable to selectively protect the amino groups or hydroxyl groups at specific positions of the raw aminoglycoside antibiotics or both with a suitable protecting group in advance.

For the selective protection of amino groups and hydroxyl groups of aminoglycoside antibiotics, several successful methods have been developed and the selective protection of relevant hydroxyl groups is used.

However, among the many amino groups present in aminoglycoside antibiotics, the selective protection of amino groups at specific positions has no significant difference in the reactivity of the amino groups, and therefore the methods currently used are difficult to implement or require some complicated manipulation.

For example, the 6'-amino group of kanamycin A is bound to only one carbon atom in an aminoglycoside molecule or to an amino group or a methyl amino group bound to a carbon atom to two or more carbon atoms in an aminoglycoside molecule. Since the reactivity is relatively higher than the amino or methylamino group bound or bonded to the carbon atom, the former amino group or methylamino group preferentially reacts with the protecting group introducing agent than the latter, and thus the former amino group or methylamino group. N-beam selectively protected

Recently, Nagabusan and its co-investigators have inactivated salts of divalent transition metals (M ⁺⁺ ) selected from copper (II), nickel (II), cobalt (II), and cadmium (II). The organic solvent is reacted with aminoglycoside antibiotics belonging to 4-0 (aminoglycosyl) -6-0- (aminoglycosyl) -2-deoxystrepamines represented by kanamycin, gentamicin and sisomycin. When the divalent transition metal cation is present in the aminoglycoside molecule, the amount of the divalent metal between the hydroxyl group and the pair of amino groups present at a specific position in the "adjacent" relationship

As such, the amino group is closed by a divalent transition metal cation.

By acting an acylating agent having an acyl group on such a complex, selective protection can be achieved with an acyl group since only amino groups which are not closed by complex bonding of divalent metal cations are mainly acylated.

This is explained below with reference to, for example, kanamycin A.

When a divalent transition metal cation selected from copper (II), nickel (II), cobalt (II), and cadmium (II) is reacted with kanamycin, the complex bonding of the divalent metal cation (M ⁺⁺ ) is as follows. As represented by formula (I), it occurs between 1-amino group and 2 "-hydroxy group and between 3" -amino group and 4 "-hydroxy group.

At least 2 moles of transition metal salt are required for 1 mole of kanamycin A in the complex. In the resulting metal complexes, the 1-amino and 3 ″ -amino groups are closed simultaneously, and then the complexes of formula (I) act an acylating agent having an acyl group which can be used as a known amino-protecting group in the general synthesis of the polypeptide. In this case, only 3-amino and 6'-amino groups which are not closed by the metal cation are mainly acylated to obtain 3,6'-di-N-acylated derivatives (American Journal 100, 5253-5254 (see 1978)). .

The inventors have found the above reported, but have further studied the interaction between various other metal cations and aminoglycoside antibiotics such as kanamycin A and kanamycin B and their derivatives. As a result, the divalent zinc cation exhibits a different function from the above-mentioned cadmium cations such as divalent nickel, cobalt and copper, and consequently has a 3'-aminoglycosyl group or a 3 "-alkylaminoglycosyl group at position 6 Regarding aminoglycoside compounds (eg kanamycin A, B, C) having oxystrept amine as a partial remedy

This expectation can be seen by the report of Nagabusan and its co-investigators in the American Journal of Chemistry, whereby kanamycin B is between 1- and 2 "-positions, between 2'- and 3'-positions and It contains three pairs of adjacent amino-hydroxyl groups between the “-and 3” -positions, but when kanamycin B reacts with a divalent metal cation, ie a zinc cation, the kanamycin B-zinc salt complex actually formed is zinc. Although complexing reactions with free 2'-amino and 3'-hydroxyl groups occur by cations, the complexing force is very low, so the 2'-amino

As another example, when kanamycin A is reacted with a zinc cation and then acylated with a benzyloxycarbonyl group (see general formula (I) above), when the zinc cation is used in an amount of 1 mole or more with respect to 1 mole of kanamycin A As the main acylation product, 3,6'-di-N-benzyloxy carbonyl kanamycin A is formed. In this case, 1,3,6 ', 3 "-tetra-N-benzyloxycarbonyl derivative of kanamycin A, unreacted raw material, and kanamycin A are simultaneously formed to some extent by this acylation reaction. It is a fact that the tri-N-benzyloxycarbonyl derivatives expected to be formed from are formed in extremely small amounts (see Example 7 below) The copper (II), nickle (N) in the specification of U.S. Pat. Salts of divalent transition metals such as (II), cobalt (II) and the like are used in an amount of at least 2 moles per mole of kanamycin A to form kanamycin A-transition metal salt complexes as indicated in the general formula (I) above. However, according to the experiments of the present invention, in the case of zinc cations, the 1-amino group of kanamycin A is 3 when used in an amount of at least 1 mole per mole of kanamycin A, unlike the four transition metal cations. "- It can be seen that an unexposed sikilsu effectively closed. According to the experiments of the present invention, the kanamycin A-nickel salt complex formed by reacting the nickel salt with kanamycin A in an amount slightly higher than 1 mole is acylated with benzyloxycarbonyl group to be 3,6'-di-N. -Benzyloxy carbonylganamycin A was obtained in very low yields but in zinc salts in good yields (see Example 7 below).

From the above fact, zinc (II) cations exhibit complex formation structures or complex stable strengths different from the cations of nickel (II), cobalt (II), copper (II) and cadmium (II). Zinc cations are also supplied in the form of zinc salts which are inexpensive and do not contaminate the surroundings.

According to the present invention, when an aminoglycoside antibiotic having a 3-aminoglycosyl group or a 2-alkylaminoglycosyl group at 6-position (which may have an aminoglycosyl group at 4-position) is reacted with a zinc cation in a suitable solvent, The amino group and the hydroxyl group at a specific position determined according to the type of aminoglycoside antibiotic are closed by complex bonding with the zinc cation at the same time, and the amino group protecting acyl used in the synthesis of the polypeptide for the aminoglycoside antibiotic and the zinc cation complex thus formed. To introduce a group, an acylating agent

According to the present invention, a zinc complex formed by reacting an aminoglycide antibiotic of deoxystreptoamine having a 3-aminoglycosyl group or a 3-alkylaminoglycosyl group at position 6 with a zinc cation in an inert organic solvent Is reacted with an acylating agent that introduces an acyl protecting group for an amino group, thereby preparing a zinc complex of an N-acylated derivative of the aminoglycoside antibiotic, which is complexed with a zinc cation, and then Provided is a method for preparing an N-protecting derivative of an aminoglycoside antibiotic selectively protected with an amino group, characterized by removing a zinc cation from a zinc complex of an N-acylated derivative with an appropriate reagent.

The method of the present invention provides that 1-N-amino acylated derivatives of kanamycins, including amikacin ("Journal of Antibiotics" 52 695-708, 1972), which have been proven to be effective antimicrobial agents. It is useful for preparing a protector in which the amino group other than the 1- and 3 "-amino groups of the raw aminoglycoside antibiotics is protected by an acyl group in the chemical synthesis of these compounds. 1-N-amino of the aminoglycoside antibiotics Acylated derivatives include kanamycin, A, kanamycin B, kanamycin C, gentamicin, sisomicin, and the like.

The above method of the present invention is described in detail below.

According to the present invention, aminoglycoside antibiotics which act on zinc cations to produce zinc complexes (called zinc complex salts) have 3-aminoglycosyl groups or 3-alkylaminoglycosyl groups as substituents on the 6-position OH group. It consists of deoxy-strep amine, which may have an aminoglycosyl group as a substituent on the hydroxyl group at the 4-position. In particular, aminoglycoside antibiotics which can be used in the present invention to form zinc cationic complexes are 6-0- (1 "-amino- or 3" -alkylamino, optionally having a 4-0- (amino-glycosyl) group. -3 "-deoxyglycosyl) -2-deoxystrept amine. Furthermore, the aminoglycoside antibiotics are 1-N-alkylaminoglycosides such as netylmycin. Examples of aminoglycoside antibiotics include kanamycin A, 6'-N-methyl-3'-deoxyganamycin A, 4'-deoxyganamycin 6'-N-methyl-4'-de Oxyganamycin A, 3 ', 4'-dideoxyganamycin A (see Japanese Patent Specification 11402/79) and 6 "-deoxy or 4", 6 "-dideoxy kanamycin A (Japanese Patent Specification No. 54733 / Kanamycin group A, kanamycin B, 3'-deoxyganamycin B (e.g. tobramycin), 4'-deoxyganamycin B, 3 ', 4'- Deoxyganamycin B (e.g. dibecacin), 3 ', 4'-dideoxy-3'-eno-ganamycin B, 6'-N-methyl-3', 4'-dideoxyganamycin B Kanamycin group B including; Kanamycin C, including kanamycin C, 3'-deoxyganamycin C, 3 ', 4'-dideoxyganamycin C; Gentamicin A, B and C; Bettamycin; There are known aminoglycosides such as sisomycin and netylmycin (eg 1-N-ethylcisomycin). Of course, the method of the present invention is known as well as unknown aminoglycoxide antibiotics

Representative examples of the aminoglycoside antibiotics to which the present invention can be applied include kanamycin A, kanamycin B, kanamycin C, and deoxy derivatives of these kanamycins represented by the following general formula (II), or 6'- N-alkyl derivatives.

(Wherein R ¹ is a hydroxyl group or an amino group, R ² and R ³ are each a hydrogen atom or a hydroxyl group, and R ⁴ is an alkylamino group containing alkyl having a hydroxyl group, an amino group or 1-4 carbon atoms, in particular a methylamino group) Indicates)

In accordance with the present invention, the free base or acid addition salt of the aminoglycoside antibiotic is dissolved or suspended in a suitable organic solvent or a hydrous organic solvent in order to react the aminoglycoside antibiotic with a zinc cation to form a complex. To this is added a suitable zinc salt in an amount of at least 1 mole per mole of aminoglycoside antibiotic. As an organic solvent that can be used, the zinc complex formed after the zinc salt is added can be at least partially dissolved in the solvent, and all conventional organic solvents can be used. However, it is desirable to avoid the use of large amounts of polar organic solvents and in particular large amounts of water, since the presence of polar solvents and water reduces the stability of the resulting aminoglycoside antibiotic-zinc cation complex, thus introducing an amino protecting group. This is because it gives a bad result for the acylation reaction.

Therefore, the solvent used for complex formation is preferably dimethyl sulfoxide having high solubility, and also hydrous dimethyl sulfoxide, dimethylformamide, hydrous dimethylformamide, mixture of dimethyl sulfoxide and dimethylformamide, tetrahydrofuran, Lower alkanols such as hydrous tetrahydrofuran, methanol, ethanol and hydrous methanol can be used.

Zinc cations are supplied in the case of zinc complex formation in the form of zinc salts. Zinc salts formed by the reaction of zinc cations with conventional inorganic or organic acids can be used in the present invention. In general, however, it is preferable to use zinc salts of weak acids such as zinc acetate because complexes containing amino groups are more stable than free salts of the free amino group and the metal complex. If a zinc salt of a strong acid is used, for example zinc chloride, the desired zinc complex is formed, but it is preferable to add a weakly alkaline salt such as sodium acetate to neutralize the medium together with the zinc salt. For the same reason, when using acid addition salts of aminoglycoside antibiotics, for example acid addition salts with strong acids such as hydrochloric acid, it is preferable to add a certain amount of base such as sodium acetate or sodium hydroxide as a neutralizing agent. In this case, however, use of unnecessary large amounts of neutralizing agents should be avoided because zinc hydroxide may precipitate, which interferes with the formation of the complex. For example, when using the tetrasulfate of the aminoglycoside antibiotic, it is preferred to add 4 moles of sodium hydroxide.

If the molar ratio of zinc salt and aminoglycoside antibiotics is equal to or greater than the same amount of zinc salt, the complex formation reaction proceeds, and it is preferable to use more moles than the complex formation so that an equilibrium reaction can occur. When using about 2,3-6 moles of zinc salt per mole of aminoglycoside, the desired yield of zinc complex can be obtained, but in practice 4-5 moles of zinc salt per mole of aminoglycoside are most preferred. Do. The time required for complex formation after addition of the zinc salt depends on the nature of the organic solvent used, which is "momentary" (function oil

An acylating agent is added to a solution or suspension containing a zinc complex of aminoglycoside antibiotics.

The acylating agent used according to the present invention is an alkanoyl group, alloyl group, alkoxycarbonyl group, alalkyloxycarbonyl group, aryloxycarbonyl group, alkylsulfonyl group It is a conventional protective reagent used for bonding an alalkyl sulfonyl group or an arylsulfonyl group. The acylating agent which can be used for this purpose is a carboxylic acid of the following general formula (IVa), an acid halide, an acid anhydride, or an active ester of this carboxylic acid (IVa); Chloroformate of the following general formula (IVb); General

(Wherein R ₅ is hydrogen, an alkyl group, in particular an alkyl group having 1-6 carbon atoms, an aryl group in particular a phenyl or alalkyl group in particular benzyl, these groups may optionally be further substituted. R ⁶ is hydrogen, an alkyl group, in particular 1 Alkyl groups, aryl groups, in particular phenyl or alalkyl, in particular phenyl alkyl, such as benzyl, having -6 carbon atoms, these groups may optionally be further substituted.)

Thus the acylation reactions for the protection of amino groups according to the invention are for example formylated, acetylated, propionylated, trifluoroacetylated, benzyloxycarbonylated, p-methoxybenzyloxycarbonylated, t -Acylated in the broad sense, including butoxycarbonylation, phenoxycarbonylation, tosylation, mesylation, and the like.

Examples of acylating agents that can be used are, for example, acetoxyformyl, p-nitrophenyl formate, acetic anhydride, acetyl chloride, propionic anhydride, p-nitrophenol ester of trifluoroacetic acid, trifluoroacetic acid ester, N-benzyloxycarbonyloxysuccinimide (representative active ester), N-benzyloxycarbonyloxyphthalimide, benzyloxycarbonyl chloride, p-methoxybenzyloxycarbonyloxy-p-nitrophenyl, t- Butoxycarbonylazide, phenoxycarbonyl chloride, tosyl chloride, mesyl chloride.

The acylating agent may be added by itself or dissolved in a solvent such as tetrahydrofuran and dimethyl sulfoxide or a mixture thereof. The amount of acylating agent added is usually used in the same mole number or slightly larger amount as the acylating agent to be reacted, but up to about three times the mole number of amino groups is optionally used. Acylating agents are added slowly over a period of 2-3 hours at a time or in small amounts, but typically over 30 minutes-1 hour. Acylation is carried out at a temperature of -20 ° C.-100 ° C., but is usually performed at a temperature from 0 ° C. to room temperature. In some cases, the temperature of acylating agent is kept low and then gradually increases when acylation is progressed. Let's do it. Usually, this acylation reaction is performed in the organic solvent used when forming an aminoglycoside antibiotic-zinc cation complex. This acylation of the zinc complex produces an N-acylated zinc complex, ie a complex of zinc cations and optionally N-acylated aminoglycoside antibiotic derivatives.

According to the above method of the present invention, after the reaction acylation described above, the zinc cation is removed from the complex of the resulting aminoglycoxide antibiotic N-acylated derivative and the zinc cation (decomposition of the zinc complex), whereby amino N-acylated protective derivatives of glycoside antibiotics are obtained.

In order to remove the zinc cation from the N-acylated zinc complex, the complex is treated with a suitable reagent to remove the zinc cation. This first method converts zinc cations into zinc sulfides, insoluble zinc compounds such as zinc hydroxide or zinc carbonate, and zinc N-acylated compounds in acylation reaction mixtures where the aminoglycoside antibiotic-zinc cation complex is acylated. The complex is dissolved or left to react with a zinc-precipitant which can then be converted from the acylation reaction mixture into a new solution in a new volume of organic solvent.

The zinc-additives that can be used in the first method include alkali metal sulfides such as hydrogen sulfide, sodium sulfide, alkaline earth metal sulfides such as ammonium sulfide and calcium sulfide, and alkali metal carbonates such as sodium carbonate or ammonium hydroxide N-acylated zinc complexes The removal of zinc cations from can be simplified by the addition of water.

According to this first method, when a zinc precipitant is added to a solution of an N-acylated zinc complex, the insoluble zinc compound formed from the zinc cation precipitates relatively quickly, so that the precipitate is filtered out. The N-acylated derivatives of the aminoglycoside antibiotics remaining in the filtrate are recovered by concentration or extraction of the solution and then purified as necessary. This tablet is useful for example Silicabell Top Chromatography.

According to the second method, the acylation reaction mixture (or other organic solvent solution) containing the N-acylated zinc complex is concentrated or concentrated to dryness to obtain an oily or solid precipitate or residue first. When a nonvolatile organic solvent such as diethyl sulfoxide is used as the N-acylation solvent, the acyl crystal reaction solution is mixed with a dilute organic solvent such as ethyl ether so that the nonvolatile organic solvent is dissolved (diluted) in the diluent. Solids or oils of N-acylated zinc complexes precipitate out. Such oils or solid precipitates or residues are usually free of (i) N-acylated derivatives of aminoglycoside antibiotics and complexes of zinc cations, and (ii) removal of organic solvents, resulting in the breakdown of some of the complexes in the complex. -Acylated derivatives, (iii) a certain amount of inorganic zinc salts formed by the breakdown of the bonds of the N-acylated zinc complexes, and (iv) a mixture of the zinc salts and the residual amount of the organic solvent used from the beginning, in excess.

This oily or solid precipitate or residue is treated by any of the following steps (a), (b) and (c).

(a) oil or solid precipitates or residues (the mixtures) are those organic liquids which have the effect of breaking the complex bonds of zinc cations in the N-acylated zinc complexes present and are obtained by dissolving free zinc salts; Organic liquids which do not dissolve the N-acylated derivatives of aminoglycoside antibiotics, mixtures of such organic liquids or mixed with hydrous organic liquids or organic liquid mixtures, decompose the N-acylated zinc complexes, zinc cations The N-acylated derivative of the desired aminoglycoside antibiotic, which is an insoluble residue, is recovered by dissolving it and dissolving it in water or a (functional) organic solvent to extract the zinc salt form. This residue is optionally dissolved by re-dissolving in an organic solvent. Polar organic solvents that can be used in this process (a) are, for example, methanol, ethanol, liquid ammonia, ethylcimine and triethylamine.

(b) oil or solid precipitates or residues (the above mixtures) are mixed with anhydrous or hydrous polar organic solvents which have the effect of breaking the complex bonds of zinc cations in the N-acylated zinc complexes, wherein zinc salts N-acylated derivatives of insoluble but desired aminoglycoside antibiotics are soluble, so polar organic solvents are added and mixed to decompose the N-acylated zinc complex to liberate N-acylated derivatives, which are added to the polar organic solvent. Dissolve, extract and separate from insoluble or residual zinc salts. In this method, a solution of N-acylated derivatives of aminoglycoside antibiotics dissolved in polar organic solvents is recovered and purified, if necessary, for example by chromatography and then concentrated to obtain the desired N-acylated derivatives.

(c) the oil or solid mixture obtained by concentrating or evaporating to dryness the extract obtained by extracting the acylation reaction solution or the acylation product derived therefrom from an organic solvent or adding a diluent liquid, when the mixture is water-soluble or partially water-soluble. The mixture is redissolved in a suitable hydrous organic solvent, and then the resulting solution is chromatographed to obtain N-acylated derivatives of zinc salts and the desired aminoglycoside antibiotics from this solution. In this chromatography process, a high polymer insoluble in a mole containing various kinds of cation exchange resins, anion exchange resins, chelate exchange resins, and functional groups capable of bonding with metals such as chitin and chitosan can be used. Types of cation-exchange resins that can be used for this purpose include those containing a carboxyl group (-COOH) and sulfonyl groups (-SO ₃ H) as exchange groups.

When using a cation exchange resin having a carboxyl functional group in the chromatography described above, the oil or solid mixture may contain a mixture of water and methanol containing 10-90 vol% water or water containing 10-90 vol% water. It is dissolved in a suitable hydrous organic solvent such as a mixture of and dioxane, and the resulting solution is charged to the column of the cation-exchange resin. The column is washed well with a higher amount of the above-mentioned organic solvent, and then developed using the above-mentioned organic solvent containing an amount of an acid or a base as an eluent. The acid is a weak organic acid like acetic acid

The concentration of acid or base in the developing solvent (eluate) is preferably 0.01 to 5% by weight. N-acylated derivatives of the desired N-acylated aminoglycoside antibiotics can be separated from the zinc cations during development because of the N-acylated derivatives for the cation-exchange resins used and the resins of the zinc cations. This is because there is a difference in adsorption force because of different binding forces. Therefore, by recovering the fraction containing the eluted N-acylated aminoglycoside separated from the zinc salt, and then concentrated to obtain the N-acylated derivative of the desired aminoglycoside antibiotics.

When the cation-exchange resin having a sulfoic acid group is used as the exchanger in the chromatography, the same structure as described above is also treated. In addition, when using weakly or strongly basic anion exchange resins in chromatography, since the N-acylated derivative has an amino group which is not acylated, the weakly basic and strongly basic anion exchange resins are ionically repulsed and are not generally adsorbed. N-acylated derivatives elute as they evolve in a suitable aqueous solvent. At this time, zinc cations are often bonded to functional groups and remain.

Since the chelate exchange resin can bind with zinc cations by its metal-chelating action, the aqueous organic solvent solution of the oil or solid precipitate or residue is first filled in a column of the chelate exchange resin, and then developed with a suitable developing solvent. Only the desired N-acylated product is eluted preferentially. Insoluble high molecular materials, such as chitin and chitosan, which are capable of binding to metals, can be used in the same manner as when chelate-exchange resins are used.

(d) The third method is to adsorb the acylation reaction solution subjected to the acylation protection reaction directly to a cation or anion-exchange resin, a chelate-exchange resin, or a column of insoluble polymer material having a metal-binding function as described above. Then, if necessary, the column is washed with a water-containing organic solvent, and the zinc cation is N-acyl by developing in the same manner as the process (c) with a water-containing organic solvent containing an acid or a base as in step (c). In addition to removal from the arc zinc complex, N-acylated derivatives of the desired aminoglycoside antibiotics can be recovered.

(e) The fourth method is an acylation containing zinc complex of N-acylated derivative and zinc cation produced in the acylation reaction when the N-acylated derivative of the desired aminoglycoside antibiotic itself is insoluble in water. The reaction solution is immediately treated with water to recover the N-acylated derivative of the desired aminoglycoside antibiotic. Examples of the case where the N-acylated derivative of this aminoglycoside antibiotic is substantially insoluble in water include 3,2 ', 6'-tri-N-benzyloxycarbonyldibecacin.

At this time, when water is added and mixed directly with the acylation reaction solution, the complex bond of the zinc complex is broken, and the N-acylated aminoglycoside derivative insoluble in water precipitates as a solid, whereas the zinc salt produced from the free zinc cation Pure N-acylated derivatives are obtained by separating from zinc salts while remaining in a silver solution. According to the method of the present invention, the acylation or amino group protection reaction is carried out on the zinc complex of the aminoglycoside antibiotic, and the complex of the mono-, di-, trinapoly-N-acylated derivative thus formed is used. Coupling with Zinc Cations

The various methods described above and the process of treating N-acylated zinc complexes with zinc cation-removing agents can be summarized as follows:

(i) The complex of the aminoglycoside antibiotic N-acylated derivative and zinc cation is separated from the acylation reaction solution before reacting with the zinc cation removal reagent.

(Ii) before reacting the complex of N-acylated derivatives and zinc cations with a zinc cation remover, extracting them with an organic solvent, evaporating the organic solvent from an acylation reaction solution, or diluting the acylation reaction solution with a diluted organic solvent. Separate from acylation reaction solution.

(Iii) The complex of zinc cation and N-acylated derivative of aminoglycoside antibiotic, once separated, is mixed with water or polar organic solvent which acts as zinc cation-removing agent. These polar free solvents should be one in which the zinc salt is dissolved but the N-acylated derivative is insoluble, or the zinc salt is insoluble but the N-acylated derivative is soluble.

(Iii) The complex of separated zinc cations and N-acylated derivatives is totally redissolved in an organic solvent containing a certain proportion of water, and then the solution is a cation-exchange resin or anion-exchange resin that acts as a zinc cation remover. Chromatography is carried out using polymers insoluble in water containing chelate-exchange resins or functional groups capable of binding to metals.

(Iii) Columns of polymers insoluble in water having a cation-exchange resin, anion-exchange resin, chelate-exchange resin, or metal-bonding function for adsorption of the acylation reaction solution to the complex of zinc cations and N-acylation derivatives. The mixture was passed directly, and then the column was developed with a hydrous organic solvent containing a certain amount of acid or base, and the eluate was collected as an oil, followed by the zinc cation containing N-acylated derivatives of the desired aminoglycoside antibiotics. Recover oil-free oil.

(Iii) When the N-acylated derivative of the desired aminoglycoside antibiotic is insoluble in water, the acylation reaction solution is treated with water directly, and the derivative is dissolved in water and precipitated separately from the remaining zinc salt.

(Iii) The acylation reaction solution is treated with hydrogen sulfide, which precipitates zinc cations as zinc sulfide, directly with alkali metal sulfides or alkaline earth metal sulfides, or with hydroxide hydroxide which precipitates zinc cations with zinc hydroxide.

According to the present invention, there is provided a method for preparing an aminoglycoside antibiotic-selective N-acylated protecting derivative, wherein the aminoglycoside antibiotic has a 6-0- (with or without 4-0- (aminoglycosyl) group. N-acylated derivatives as described above having 3 "amino- or 3" -alkylamino-3 "-deoxyglycosyl) -2-deoxy-streptamine and some amine groups optionally protected with amino-protected acyl groups Consisting of

(a) A zinc cation complex and an inert organic solvent of an aminoglycoside antibiotic formed by the reaction of an aminoglycoside antibiotic and a zinc salt with an acylating agent having an acyl group introduced into the aminoglycoside antibiotic N- while reacting at a temperature of -20 ° C to 100 ° C, preferably at room temperature, in the presence of to acylate unbonded amino groups present in the aminoglycoside antibiotic-zincionic complex Preparing a complex of aminoglycoside antibiotic-zinc cations.

(b) the complex of N-aminoglycoxide antibiotic-zinc cation in water or in anhydrous organic solvent, liquid ammonia, ethylamine and triethylamine, or in hydrogen sulfide, alkali metal sulfide or alkaline earth metal sulfide or water Ammonium hydroxide, or cation-exchange resins containing carboxylic acid or sulfonic acid functional groups, or anion-exchange resins containing ammonium groups, or chelate-exchange resins containing metal-chelate functional groups, or metal-linked groups It comprises a step of preparing a selective N-acylation protection derivative of aminoglycoside antibiotics by removing the zinc cation from the complex by heating or cooling with chitin or chitosan and reacting at room temperature with an insoluble polymer insoluble containing.

As described above in the method of the present invention, an N-acylated derivative of an aminoglycoside antibiotic which mainly protects other amino or alkylamino groups other than 1-amino and 3 "-amino groups or alkylamino groups by acyl groups is obtained.

N-acylated derivatives of the aminoglycoside antibiotics thus obtained as the method of the present invention are known as α-hydroxy-W-aminoalkanoic acid by known methods such as those described in, for example, US Pat. Nos. 3,781,268 and 3,939,143. 1-N-acylation, followed by semi-synthetic 1-N-acylated aminoglycoside antibiotics known as useful antimicrobial agents by removing residual amino-protecting groups from the resulting 1-N-acylated products.

The use of kanamycin A as a starting material in the synthesis of 1-N-acylated aminoglycoside antibiotics is described. Since kanamycin A is closed by complexation with zinc cations with the 1-amino group and the 3 "-amino group, kanamycin A-zinc complex acts according to the present invention by acting a suitable acylating agent or other type of amino group protecting agent. The amino group and the 6'-amino group may be protected by the acyl group of the acylating agent or other amino protecting group, followed by removal of the zinc cation from the N-acylated kanamycin A-zincionic complex, followed by When reacted with an acylation having an acylation to be introduced into the 1-amino group, this acylation group reacts only with free 1-amino and 3 ″ -amino groups. Since the 1-amino group is usually higher in reactivity than the 3 ″ -amino group, the desired 1-N-acylated kanamycin derivative is obtained in higher yield than the 3 ″ -N-acylated kanamycin A derivative. This will result in higher yield of acylating agent to the desired 1-N-acylkanamycin A in higher yield compared to when the kanamycin A or 6'-amino group was reacted directly with the previously protected kanamycin A. In comparison, the desired yield of 1-N-acylganamycin A is obtained. If kanamycin is reacted with an acylating agent without N-protection, a mixed N-acylation product is formed that contains a very small proportion (usually 1% -s%) of the desired 1-N-acylation product.

When the method of the present invention is applied to the preparation of kanamycin of the general formula (III), N or a part of or all of the amino groups other than the 1- and 3 ″ -amino groups of the used kanamycin is represented by the following general formula (V). Obtain an acylated derivative.

기, -NHCO.OR⁵기,

기, -NHSO₂R⁶기나

기이며; R²와 R³는 각각 일반식(Ⅲ)에서 정의된 의미와 같고,R⁷은 -COR⁵기, -CO.OR⁵또는 -SO₂R⁶기이고, R⁵와 R⁶는 각각 일반식(Ⅳa)-(Ⅳf)에서와 같으며, R⁸는 1-4개의 탄소원자를 갖는 알킬기이다).[In the formula R ¹ _a is a hydroxy group, an amino group (NH _2), - NHCOR ⁵ group, a ⁵ -NHCO.OR group or -NHSO ₂ R ⁶ group, R ^4a is a hydroxy group, -NHCOR ⁵ group,

-NHCO.OR ⁵ groups,

Group, -NHSO ₂ R ⁶ group

Group; R ² and R ³ are the same as defined in general formula (III), R ⁷ is a -COR ⁵ group, -CO.OR ⁵ or -SO ₂ R ⁶ group, and R ⁵ and R ⁶ are each general formula As in (IVa)-(IVf), R ⁸ is an alkyl group having 1-4 carbon atoms).

When the method of the present invention is applied to kanamycins, n-protected kanamycin derivatives of general formula (V) in which both amino groups and alkylamino groups other than the 1- and 3 "-positions of the kanamycin molecule are protected by the above-mentioned group. Reducing the number of moles of acylating agent used or shortening the reaction time if the protective amino group to be introduced is large in size (e.g. t-butoxycarbonyl group) or if the protective acyl group to be introduced is of ordinary size In this case, the N-protected kanamycin derivative is less protected by the number of acyl groups introduced, since the 6'-amino group or 6'-alkylamino group is more reactive than other amino groups. Only alkylamino groups result in protected substances N-acylated kanamycin derivatives of general formula (V) are useful intermediates for the semi-synthesis of various kanamycin derivatives. C. The 1-amino group of Compound (V) is acylated with α-hydroxyl-W-aminoalkanoic acid, and then semi-synthetic 1-N effective for resistant bacteria by removing protecting groups of amino or alkylamino groups other than this 1-acyl group. It is of great value as an intermediate of compound (V) in chemical aptitude, including the manufacturing process of acylated aminoglycoside antibiotics.

For example, when linking an acyl group to compound (V), that is, (S) -4-benzyloxycarbonylamino-2-hydroxybutyl group, compound (V) can be reacted in a suitable organic solvent such as hydrous tetrahydrofuran. The 1-N-acylated product is formed by reacting with substituted butyric acid or an active ester such as N-hydroxy succinimide ester, N-hydroxyphthalimide ester or p-nitrophenol ester. Next, the benzyloxycarbonyl group and the protecting group (R) of the general formula (V) from the 1-N-acylated product are subjected to conventional methods, for example hydrolysis by acid or base, metal

(In the formula, R ¹ , R ² , R ³ and R ⁴ have the same meaning as in general formula (III).

Example 1

Preparation of 3,6'-di-N-benzyloxycarbonyl kanamycin A

의 냉각된 용액(0℃)을 약1시간 동안 서서히 첨가한다. 반응용액을 실온에서 4시간 방치시킨다. 이때 가나마이신 A의 아연착체는 벤질옥시카르보닐화(본 발명의 첫번재 형태에 따른 아실화)가 진행된다.(i) 2.0 g (4.13 mmol) kanamycin A (freebase) is suspended in a mixture of dimethyl sulfoxide (50 ml) and tetrahydrofuran (20 ml) and 4 g (18.1 ml) zinc acetate (II) in suspension. Dihydrate is added and then stirred at room temperature until the reaction mixture is homogeneous. Zinc complexes of kanamycin A are formed and dissolved for 4-5 hours. 2.37 g (9.5 mmol) N-benzyloxycarbonyloxy dissolved in a mixture (40 ml) of tetrahydrofuran-dimethylsulfoxide (1: 1 by volume) in the resulting solution was cooled to 0 ° C. Succinimide

Of cooled solution (0 ° C.) is added slowly for about 1 hour. The reaction solution is left at room temperature for 4 hours. At this time, the zinc complex of kanamycin A undergoes benzyloxycarbonylation (acylation according to the first form of the present invention).

The sample taken from the reaction solution thus obtained was subjected to silica gel thin layer chromatography using the bottom liquid phase of the mixture of chloroform-methanol-28% aqueous ammonia (1: 1: 1 by volume) as a developing solvent. ) Appears at Rf = 0.23, and two or three small spots appear as upper by-products.

(ii) The reaction solution was poured into 50 ml of ethyl ether, and the precipitated oil was washed several times with a higher volume of ethyl ether to obtain 8.8 g of thick syrup.

(iii) The zinc cation is removed from the syrup material (which consists of zinc complexes) using the following methods (A)-(J): (A) A weakly acidic cation-exchange resin having a carboxyl group (-COOH) as a functional group (US Industrial "Amberite" GG50 resin (H ⁺ form) commercially available from Hohm & Haas Company.

60 ml of Amberlite CG50 resin (H ⁺ form) was first treated sufficiently with a mixture of water-dioxane (2: 1), then charged into a column and dissolved in 20 ml of water-dioxane (1: 1). 1 g of the syrup material solution is charged to the column and then developed with water-dioxane (2: 1) containing 1% acetic acid. The eluate is recovered as an oil, and the desired 3,6'-di-N-benzyloxycarbonyl kanamycin A, which is positive for the ninhydrin reaction, is eluted first, followed by elution of zinc acetate which is specialized for color development by diphenylcarbazide. Let's do it. The fractions containing the desired product are combined together and concentrated to dryness. Washing the concentrate with ethyl ether gave 340 mg (81%) of 3,6'-di-N-benzyloxycarbonyl kanamycin A as a colorless solid.

+76°(c1, 물-디메틸포름아미드, 1 : 2)

+ 76 ° (c1, water-dimethylformamide, 1: 2)

Elemental Analysis C ₃₄ H ₄₈ N ₄ O ₁₅ .2CH ₃ CO ₂ HH ₂ O

Theoretic value: C, 51.23; H, 6.56; N, 6.29%

Anal: C, 51.02; H, 6.71; N, 6.22%

형태)로서 시판되는)를 사용한 방법.(B) Weak cation-exchange resins having carboxylate groups as functional groups ("Amberite" CG50 resins from Rohm & Haas Company (

Commercially available).

형태)의 컬럼에 채우고, 0-0.1N 암모니아를 함유하는 물-디옥산(1 : 1)로 경사 전개한다. 아연 양이온은 용출되지 않으나, 원하는 생성물인 3,6'-디-N-벤질옥시카르보닐 가나마이신 A가 용출된다. 원하는 벤질옥시카르보닐화 생성물을 함유하는 용출액의 유분을 농축건조하면 무색고체인 328mg(89%)의 원하는 생성물을 얻는다.1 g of the syrup-like material obtained in Example 1 (ii) was dissolved in 20 ml of water-dioxane (1: 1) and the solution was dissolved in 60 ml of Amberlite GC50 resin (

Column) and gradient development with water-dioxane (1: 1) containing 0-0.1 N ammonia. The zinc cation does not elute, but the desired product 3,6'-di-N-benzyloxycarbonyl kanamycin A elutes. Concentration of the fraction of the eluate containing the desired benzyloxycarbonylated product yields 328 mg (89%) of the desired product as a colorless solid.

+86°(c1, 물-디메틸포름아마이드 = 1 : 2)

+ 86 ° (c1, water-dimethylformamide = 1: 2)

H₂CO₃ Elemental Analysis C ₃₄ H ₄₈ N ₄ O ₁₅ ·

H ₂ CO ₃

Theoretic value: C, 52.87; H, 6.30; N, 7.15%

Anal: C, 52.50; H, 6.30; N, 7.00%

(C) A method using a cation-exchange resin having a strongly acidic functional group (-SO ₃ H) (commercially available from Dow Chemicals as "Doughx" 50 W x 2 resin).

Example 1 (ii), filled with 30 ml of Dow's 50W × 2 resin (H ⁺ form) treated with water-dioxane (2: 1), in 20 ml of water-dioxane (2: 1) Fill a solution of 1 g of syrup-like material obtained from. The column is washed with water-dioxane (2: 1) until the eluate is neutral and then decanted with water-dioxane (2: 1) containing 0-1-N-ammonia. The eluate fractions containing the desired 3,6'-di-N-belzyloxycarbonyl kanamycin A were concentrated to dryness under reduced pressure to give 311 mg (84%) of a white solid as obtained in Example 1 (iii).

(D) Another method using Dow's 50W × 2.

30 ml of Dow's 50W × 2 resin, which was previously treated with water-methanol (3: 1), with a solution of 1 g of the serum substance obtained in Example 1 (ii) dissolved in 20 ml of water-methanol (3: 1) H ⁺ form) is placed in a column, washed well with water-ethanol (3: 1) and then decanted with water-ethanol (3: 1) containing 0-6N hydrochloric acid. The active base containing the desired 3,6'-di-N-benzyloxycarbonyl kanamycin A is collected in a strong base ion-exchange resin, e.g. Dow's 1x2, in an amount sufficient to make the mixture weakly acidic. It is mixed with the resin (OH form).

The mixture is filtered and the filtrate is concentrated to dryness to give 285 mg (72%) of the desired product as this hydrochloride.

+79°(c1, 물-디메틸포름 아마이드 = 1 : 2)

+ 79 ° (c1, water-dimethylformamide = 1: 2)

(E) A method using an anion-exchange resin having a strong basic group and a tetravalent ammonium group (a commercially available Dowks 1 × 2 resin available from Dow Chemical Company) as a functional group.

30 ml of Dow's 1 × 2 resin (OH) pretreated with a water-dioxane (1: 1) solution of 1 g of syrup-like material obtained in Example 1 (ii) dissolved in water-dioxane (1: 1) Form) and then develop relatively rapidly as water-dioxane (1: 1). Eluent fractions containing the desired product were collected and concentrated to dryness to yield 305 mg (84%) of a colorless solid as in Example 1 (iii) (B).

(F) A method using an anion-exchange resin (commercially available from Dow Chemicals as WGR resin) having weak basicity as a functional group.

1 g of syrup-like material obtained in Example 1 (ii) was dissolved in 20 ml of water-dioxane (1: 1) and 50 ml of Dow's WGR resin (prepared with water-dioxane (2: 1)) Base) column and then developed with water-dioxane (2: 1). The desired 3,6'-di-N-benzyloxycarbonyl kanamycin A is eluted with trace amounts of zinc cations.

These eluates are mixed together and concentrated to dryness to yield 450 mg of a colorless solid. This solid was used as starting material for the preparation of 1-N-((S) -4-amino-2-hydroxybutyryl) ganamycin A according to the 1-N-acylation method of Example 31 described below. Used directly, where traces of zinc cations remaining in the solid starting material do not adversely affect the acylation reactions included in Example 31.

(G) A method using chelate exchange resin (commercially available as Dow's A ₁ resin from Dow Chemical Company) having a weakly acidic group as a functional group.

A solution of 1 g of syrup obtained in Example 1 (ii) dissolved in water-dioxane (1: 1) was pretreated with 50 ml of Dox A, pretreated with water-dioxane (1: 1) containing 1% ammonia. It is introduced into a column of resin and then decanted with a mixture of water-dioxane (1: 1) containing 0-1-N-ammonia. Only the portion containing the desired 3,6'-N-benzyloxycarbonyl kanamycin A is eluted. Concentration to dryness gives 272 mg (74%) of the desired product as a white solid.

(H) A method using chitosan (a polymer that is commercially available from Toko Kasai Co., Ltd., Japan, and insoluble in water containing a functional group capable of bonding with a metal).

형태)의 컬럼에 넣는다. 칼럼을 물-디옥산(1 : 1)로 잘 세척한 다음 0-0.1N암모니아를 함유하는 물-디옥산(1 : 1)으로 경사 전개시킨후, 닌히드린 반응에 민감한 이들 유분을 함께 결합시키고, 농축 건조하면 실시예 1(iii)(B)에서 얻은 것과동일한 30mg(82%)의 무색 고체를 얻는다.A solution of 1 g of syrup-like material obtained in Example 1 (ii), in which 100 ml of chitosan was sufficiently pretreated with water-methanol (3: 1), filled into a column and dissolved in water-methanol (3: 1), was prepared. When developed with methanol (3: 1), the desired 3,6'-di-N-benzyloxycarbonyl kanamycin A elutes first and zinc acetate elutes later. The former is concentrated to dryness, leaving a residue that is aberite CG50 resin pretreated with water-dioxane (1: 1) (

Form). The column was washed well with water-dioxane (1: 1) and then decanted with water-dioxane (1: 1) containing 0-0.1 N ammonia, then these oils sensitive to the ninhydrin reaction were bound together. Concentration and drying give 30 mg (82%) of a colorless solid identical to that obtained in Example 1 (iii) (B).

(I) using a polymer having a carboxyl functional group (commercially available as "CM- Sephadex" C-25, an ion-exchange gel-filter composed of a carboxymethyl-substituted test gel made by Parmasica Fine Chemicals, Sweden) Way.

40 ml of CM-Sephadex C-25, sufficiently treated with water-dioxane (1: 1), of a solution of 1 g of syrup-like material obtained in Example 1 (ii) dissolved in water-dioxane (1: 1) Packed into a column of (T8-6), washed the column with 200 ml of water-dioxane (1: 1) and then decanted with water-dioxane (1: 1) containing 0-0.1N ammonia Deploy it.

The zinc cation does not elute from the column but only the desired 3,6'-di-N-belzyloxycarbonyl kanamycin A elutes. Concentration and drying of the eluate gave 303 mg (82%) of a colorless solid identical to that of Example 1 (iii) (B).

(J) A method using hydrogen sulfide as a zinc precipitant.

1 g of the granule obtained in Example 1 (ii) is dissolved in 20 ml of water-methanol (1: 1), to which ammonia water is added, and then a sufficient amount of hydrogen sulfide is introduced. The reaction mussels containing zinc sulfide precipitates are filtered through a glass filter filled with a "cerite" filter, and the filtrate is concentrated under reduced pressure to leave syrup material, which is washed with ethyl ether to leave a solid residue. This residue was dissolved in a constant volume of water-dioxane (1: 1) and the solution was dissolved in water-dioxane (1 :) as a developing solvent, and 30 ml of amberanite IRA900 (OH form, strong base resin, Rohm) was used. Chromatography on a column of & Haas). The fractions containing 3,6'-di-N-benzyloxycarbonyl kanamycin A are mixed together and concentrated to dryness to yield 235 mg (64%) of a colorless solid as in Example 1 (iii) (B). .

Example 2

Preparation of 3,6'-di-N-benzyloxycarbonyl kanamycin A

500 mg (1.03 mmol) of kanamycin A (freebase) are suspended in 15 ml of dimethylsulfoxide followed by addition of 420 mg (3.09 mmol) of zinc chloride and 840 mg (6.18 mmol) of sodium acetate trihydrate. .

)용액을 약 한시간 이상 첨가한다. 생성된 혼합물을 실온에서 4시간 방치한다. 다음에, 반응 혼합물을 실시예 1(ⅱ)과 (ⅲ)(Ⅰ)에 기술된 바와 동일한 방법으로 처리하면, 무색고체의 형태로서 598mg(74%)의 3,6'-디-N-벤질옥시카르보닐 가나마이신 AAfter stirring the mixture for 10 hours, 675 mg (2.27 mmol) N-benzyloxycarbonyl oxyphthalimide (dissolved in 10 ml of dimethyl sulfoxide in a mixture containing the formed kanamycin A-zinc complex)

Add the solution for about an hour or more. The resulting mixture is left for 4 hours at room temperature. Next, the reaction mixture was treated in the same manner as described in Examples 1 (ii) and (iii) (I) to give 598 mg (74%) of 3,6'-di-N-benzyl as a colorless solid. Oxycarbonyl kanamycin A

Example 3

Preparation of 3,6'-di-N-benzyloxycarbonyl kanamycin A

In 15 ml of dimethyl sulfoxide, 600 mg (0.95 mol) of kanamycin A4 hydrochloride and 150 mg (3.8 mmol) of sodium hydroxide were stirred for 1 hour, followed by 1 g (4.55 mol) of zinc acetate dihydrate. After the addition, stirring was continued for another 5 hours. To a mixture containing the formed kanamycin A-zinc complex, a solution of 545 mg (2.2 mmol) of N-belzyloxycarbonyloxysuccinimide dissolved in 5 ml of dimethylsulfoxide-tetrahydrofuran (1: 1) was added. Add over minutes. The resulting mixture was stirred at room temperature overnight, and then ethyl ether was added thereto to give N-

Example 4

Preparation of 3,6'-di-N-benzyloxycarbonylganamycin A

(i) 500 mg (1.03 m mole) of kanamycin A (free base) is dissolved in 20 ml of water-dimethylsulfoxide (1: 9) mixture and 1 g (4.55 m) of zinc acetate dihydrate is added to this solution. Then 590 mg (2.4 mmol) of N-benzyloxycarbonyloxysuccinamide is added. After the mixture was left at room temperature for one night, a large amount of ethyl ether was added to the mixture, the precipitated water layer was separated and washed several times with ethyl ether to obtain a concentrated syrup layer.

형태)의 컬럼에 넣고, 컬럼을 물-디옥산(1 : 1)의 혼합물로 잘 세척한 다음, 0-0.1N 암모니아를 함유하는 물-디옥산(1 : 1)로 경사 전개시킨다. 원하는 생성물을 함유하는 부분을 함께 혼합하고 농축 건조하면 실시예 1(iii)(B)에서 얻는 것과 동일한 494mg(51%)의 무색고체를 얻는다.(ii) The syrup material thus obtained is dissolved in water-methanol (3: 1) and the solution is placed in a column of 200 ml chitosan. The column is developed with water-methanol (3: 1), the eluate is collected as an oil, the oils positive for the ninhydrin reaction are combined together and concentrated. Concentrate the Amberlite CG50 Resin (

Form), and the column is washed well with a mixture of water-dioxane (1: 1) and then decanted with water-dioxane (1: 1) containing 0-0.1N ammonia. The portions containing the desired product are mixed together and concentrated to dryness to give 494 mg (51%) of a colorless solid as obtained in Example 1 (iii) (B).

Example 5

Preparation of 3,6'-di-N-benzyloxycarbonyl kanamycin A

500 mg (1.03 mmol) of kanamycin A (freebase) are dissolved in 15 ml of water-tetrahydroputane (1: 3) mixture, to which 1 g (4.55 mmol) zinc acetate dihydrate is added, followed by 590 mg (2.4 mmol) N-benzyloxycarbonyl oxysuccinimide is added. The mixture was left at room temperature overnight, the reaction mixture was concentrated under reduced pressure, and the residue was placed in a column of 200 ml of chitosan, and the effluent from the column was treated in the same manner as in Example 4 (ii) to give 414 mg (51). The compound of%) is obtained as a colorless solid.

Example 6

Preparation of 3,6'-di-N-benzyloxycarbonyl kanamycin A

(i) 500 mg (1.03 m mole) of kanamycin A (freebase) is dissolved in 15 ml of water-methanol (1: 7) mixture, to which 1.5 g (6.8 m) zinc acetate dihydrate is added, After addition of 590 mg (2.4 mmol) of N-benzyloxycarbonyloxysuccinimide dissolved in 7 ml of tetrahydrobutane, the mixture was left at room temperature for one night, and the reaction mixture was concentrated under reduced pressure. 200 ml of chitosan was added to the column and the effluent from the column was treated in the same manner as in Example 4 (ii) to give 442 mg (55%) of the compound as a colorless solid.

Example 7

Preparation of 3,6'-di-N-benzyloxycarbonyl kanamycin A

500 mg (1.03 mmol) kanamycin A (freebase) are suspended in 20 ml of dimethylsulfoxide and 272 mg (1.24 mmol) zinc acetate dihydrate are added to the suspension. The mixture was stirred at room temperature for 10 hours to give a clear solution, in which 540 mg (2.17 mmol) of N-benzyloxycarbonyloxysuccinimide was added dropwise over about 2 hours.

Next, the resultant mixture was left at room temperature for one night, and then a large amount of ethyl ether was added to the solution, and the precipitated oily substance was collected and washed several times with ethyl ether to obtain a syrupy substance.

A sample taken from the syrup-like material is called silica gel thin layer chromatography using chloroform-methanol-28% ammonia water (1: 1: 1 in the lower layer) as a developing solvent.

Weak spot of 1,3,6 ', 3 "-tetra-N-benzyloxycarbonyl kanamycin A at -Rf 0.4 (color developed by spraying sulfuric acid on a laminate followed by heating);

Spot weak at Rf 0.28;

Strong spot of the desired product at -Rf 0.23, namely 3,6'-di-N-benzyloxycarbonyl kanamycin A;

Weak spot of 6'-N-benzyloxycarbonyl kanamycin A at -Rf 0.12;

Extremely weak spot of unreacted kanamycin A at -Rf 0.

However, no spots corresponding to tri-N-benzyloxycarbonyl kanamycin A appearing in Rf 0.28-0.4 were observed.

100 ml of CM- Sephadex C-25 resin (NH ₄ ⁺ form), in which the above syrup-like substance was dissolved in water-dioxane (1: 1) and the solution was pretreated with water-dioxane (1: 1). The column was then subjected to the same process as described in Example 1 (iii), the zinc cation was removed and the desired product separated from the other product was separated to give a colorless solid of 412 mg (51%). To the present compound.

For comparison, the above reaction was repeated but the zinc acetate dihydrate was replaced with 308 mg (1.24 mmol) of nickel acetate (II) acetate tetrahydrate, resulting in the desired 3,6'-di-N-benzyloxy as a colorless conductor. Carbonyl kanamycin A is obtained in a low yield of 59 mg (7.3%).

Example 8

Preparation of 3,6'-di-N- (p-methoxybenzyloxycarbonyl) kanamycin A

500 mg (1.03 mmol) kanamycin A (freebase) are suspended in 12 ml dimethylsulfoxide and 1 g (4.55 mmol) zinc acetate dihydrate is added to this suspension. The mixture was stirred at room temperature until a homogeneous solution, then 789 mg (2.6 mmol) of p-methoxycarbonyl benzoxy p-nitrophenyl ester (p-CH ₃ OC ₆ ) dissolved in 10 ml of dimethylsulfoxide H ₄ CH ₂ OCOOC ₆ H ₄ p-NO ₂ ) solution was added over about 30 minutes, left at room temperature for 1 night, and then as in Example 1 (ii) and (iii) (B).

+87°(c1, 물-디메틸설폭사이드=1:2)

+ 87 ° (c1, water-dimethylsulfoxide = 1: 2)

H₂CO₃ Elemental Analysis C ₃₆ H ₅₂ N ₄ O ₁₇ ·

H ₂ CO ₃

Theoretical: C, 51.95; H, 6.33; N, 6.64%

Anal: C, 51.56; H, 6. 41; N, 6.53%

Example 9

Preparation of 6'-N- (t-butoxycarbonyl) ganamycin A

Except for replacing p-methoxycarbenzoxy p-nitrophenyl ester with 220 mg (1.54 mmol) of t-butoxycarbonylazide, the compound was obtained as a colorless solid according to the same procedure as described in Example 8. Get

Yield = 627 mg

+96°(c1, 물-디메틸포름 아미드=1 : 2)

+ 96 ° (c1, water-dimethylformamide = 1: 2)

Example 10

Preparation of 3,6 (-di-N-trifluoroacetyl kanamycin A

500 mg (1.03 mol) of kanamycin A (freebase) are suspended in 12 ml of dimethylsulfoxide and 1 g (4.55 mmol) zinc acetate dihydrate is added to this suspension.

The mixture was stirred at room temperature until a homogeneous solution was added thereto, followed by addition of 1.2 g (5.1 mmol) of p-nitrophenol ester solution of trifluoroacetic acid dissolved in 10 ml of dimethylsulfoxide, followed by one night at room temperature. After standing, treatment with ethyl ether as described in Example 1 (ii) followed by treatment of the ether-insoluble syrup material in the same manner as in Example 1 (iii) (A) yielded 590 mg (70%) of a colorless solid. To the present compound.

+81°(c1, 물-디메틸포름아미드=1 : 2)

+ 81 ° (c1, water-dimethylformamide = 1: 2)

Elemental Analysis C ₂₂ H ₃₄ N ₄ O ₁₃ · 2CH ₃ CO ₂ HH ₂ O

Theoretical: C, 38.33; H, 5. 44; N, 6.88%, F, 13.99%

Anal: C, 38.03; H, 5. 48; N, 6.54%

Example 11

Preparation of 3,6'-di-N-phenoxycarbonyl kanamycin A

500 mg (1.03 mmol) kanamycin A (freebase) was suspended in a mixture of dimethyl sulfoxide (15 ml) and tetrahydrofuran (5 ml), and 1 g (4.55 mmol) zinc acetate dihydrate was added to the suspension. Then, the reaction mixture is stirred at room temperature until uniform. The resulting solution is cooled to 10 ° C., and then slowly added 400 ml (2.5 mmol) of phenoxycarbonyl chloride (C ₆ H ₅ OCOC) in a cooled solution (0 ° C.) dissolved in 3 ml of tetrahydrofuran. . The reaction solution was allowed to reach room temperature over 1 hour, and then left at this temperature for 3 hours, followed by reaction mixture.

+73°(cl, 물-디메틸설폭사이드=1:2)

+ 73 ° (cl, water-dimethylsulfoxide = 1: 2)

Elemental analysis: C ₃₂ H ₄₄ N ₄ O ₁₅ · 2CH ₃ CO ₂ HH ₂ O

Theoretical: C, 50.11; H, 6. 31; N, 6.49%

Anal: C, 49.77; H, 6. 60; N, 6.11%

Example 12

Preparation of 3,6'-di-N-acetyl kanamycin A

Example 1 (iii) (A) A reaction mixture obtained in the same manner as in Example 1 except for using 260 mg (2.6 mmol of acetic anhydride) instead of p-methoxycarbenzoxy p-nitrophenyl ester was used. Treatment in the same manner as described above yields 525 mg (72%) of the present compound as a colorless solid.

+93°(c1, 물-디메틸포름아미드=1 : 2)

+ 93 ° (c1, water-dimethylformamide = 1: 2)

Elemental Analysis C ₂₂ H ₄₀ N ₄ O ₁₃ · 2CH ₃ CO ₂ HH ₂ O

Theoretic value: C, 44.19; H, 7.13; N, 7.93%

Anal: C, 44.20; H, 7.07; N, 7.85%

Example 13

Preparation of 3,6'-di-N-formylganamycin A

500 g (1.03 mmol) of kanamycin A (freebase) were suspended in 12 ml of dimethylsulfoxide, 1 g (4.55 m) of zinc acetate dihydrate was added to the suspension, followed by stirring at room temperature until a uniform solution was obtained. Then 690 mg (4.12 mmol) of p-nitrophenylformate (OHCOC ₆ H ₄ -P-NO ₂ ) are added. The resulting mixture is left at room temperature for one night and then treated in the same manner as in Example 1 (iii) (B). The positive components of the ninhydrin reaction are mixed together, carbon dioxide gas is introduced, and concentrated to dryness to obtain 430 mg (67%) of the present compound as a colorless solid.

+101°(c1, 물)

+ 101 ° (c1, water)

Elemental analysis _{C 20 H 36 N 4 O 13} · H 2 CO 3 HH 2 O

Theoretical: C, 40.64; H, 6. 50; N, 9.03%

Anal: C, 40.43; H, 6. 47; N, 8.83%

Example 14

Preparation of 3,6'-di-N-tosylganamycin A

500 mg (1.03 mmol) of kanamycin A (freebase) are suspended in 15 ml of dimethylsulfoxide and 1 g (4.55 mmol) zinc acetate dihydrate is added to the suspension. Stir at room temperature until the reaction mixture is uniform, and slowly add 400 mg (2.1 mmol) of tosyl chloride solution dissolved in 7 ml of tetrahydrofuran. The resulting mixture was left at room temperature for 1 hour, and then 200 mg of tosyl chloride dissolved in 3.5 ml of tetrahydrofuran was added again, and the reaction mixture was allowed to stand for another 2 hours. Examples 1 (ii) and (iii) Treatment in the same manner as described in (A) yields 270 mg (28%) of the present compound as a colorless solid.

+68°(c1, 물-디메틸포름아미드=1 : 2)

+ 68 ° (c1, water-dimethylformamide = 1: 2)

Elemental analysis _{C 32 H 48 N 4 O 15} S 2 · 2H 3 CO 2 HH 2 O

Theoretic value: C, 46.44; H, 6. 28; N, 6.02; S, 6.89%

Anal: C, 46.31; H, 5.98; N, 6.31; S; 6.55%

When the reaction was repeated without using zinc acetate, no colorless solid product was recovered.

Example 15

Preparation of 3,6'-di-N-benzyloxycarbonyl-6'-N-methylganamycin A

500 mg (1.0 mmol) of 6'-N-methylganamycin A (freebase) are suspended in 12 ml of dimethylsulfoxide, and 1 g (4.55 m5 mlmol) of zinc acetate dianhydride is added to the suspension, followed by uniformity. After stirring at room temperature until a mixture, 550 mg (2.2 mmol) of N-benzyloxycarbonyloxysuccinimide solution dissolved in 5 ml of dimethyl sulfoxide-tetrahydrofuran (1: 1) was mixed for 30 minutes. Add over. The resulting mixture was left at room temperature for one night and then treated in the same manner as in Example 1 (ii) alc (iii) (A) to afford 720 mg (79%) of the present compound as a colorless solid.

+74°(c1, 물-디메틸포름아미드=1 : 2)

+ 74 ° (c1, water-dimethylformamide = 1: 2)

Treatment of this compound by a method analogous to that described in Example 31 below yields 1-N-((S) -4-amino-2-hydroxybutyryl) -6'-N-methylganamycin A .

Example 16

Preparation of 3,6'-N-benzyloxycarbonyl-3'-deoxyganamycin A

The same procedure as in Example 15 was repeated but 500 mg (1.07 mmol) of 3'-deoxyganamycin A (free base) was used as a starting material and 610 mg (2.45 mmol) of N-benzyloxycarbonyloxysuccinate. By using mead, 765 mg (82%) of the present compound in colorless solid form are obtained.

+76°(c1, 물-디메틸포름아미드=1 : 2)

+ 76 ° (c1, water-dimethylformamide = 1: 2)

Elemental Analysis C ₃₄ H ₄₈ N ₄ O ₁₄ · 2CH. ₃ CO ₂ HH ₂ O

Theoretical: C, 52.16; H, 6.68; N, 6.40%

Anal: C, 51.99; H, 6.75; N, 6.20%

The compound is treated by a method analogous to that described in Example 31 to obtain 1-N-((S) -4-amino-2-hydroxybutyryl) -3'-deoxyganamycin A.

Example 17

Preparation of 3,6'-di-N-benzyloxycarbonyl-3'-deoxy-6'-N-methyl kanamycin A

Repeat the process of Example 15, starting with 500 mg (1.04 mmol) of 3'-deoxy-6'-N-methyl kanamycin A (freebase) and 595 mg (2.4 mmol) of N-benzyl Using oxycarbonyloxysuccinamide yields 737 mg (80%) of this compound.

+73°(c1, 물-디메틸포름아미드=1 : 2)

+ 73 ° (c1, water-dimethylformamide = 1: 2)

Treatment of this compound in a manner similar to that described in Example 31 resulted in 1-N-((S) -4-amino-2-hydroxybutyryl) -3'-deoxy-6'-N-methyl kanamycin Get A

Example 18

Preparation of 3,6'-di-N-benzyloxycarbonyl-4'-deoxyganamycin A

500 mg (1.07 mmol) of 4'-deoxy, except slowly adding 580 mg (2.3 mmol) of N-benzyloxycarbonyloxysuccinamide dissolved in 4 ml of dimethyl sulfoxide to a uniform solution over 1 hour Example 15 using kanamycin (free base) (see "Journl of Antibiotics vol. 27, pp 838 849 (1974)" Bulletin of Chemical Society of Japan "vol. 50, pp 2362 2368 (1977)) as a starting material In the same manner as in the following to obtain 666 mg (71%) of the present compound as a colorless solid.

+77°(c1, 물-디메틸포름아미드=1 : 2)

+ 77 ° (c1, water-dimethylformamide = 1: 2)

Elemental Analysis C ₃₄ H ₄₈ N ₄ O ₁₈ · 2CH ₃ CO ₂ HH ₂ O

Theoretical: C, 52.16; H, 6.68; N, 6.40%

Anal: C, 51.77; H, 6.79; N, 6.31%

Example 19

Preparation of 3,2'6'-tri-N-benzyloxycarbonyl kanamycin B

500 mg (1.03 mmol) of kanamycin B (freebase) are suspended in a mixture of 12 ml of dimethylsulfoxide and 4 ml of tetrahydrofuran and 1 g (4.55 mmol) of zinc acetate dihydrate is added to the suspension and homogeneous. Stir at room temperature until solution, then cool to 0 ° C. and 825 mg 3.3 mmol of N-benzyloxycarbonyloxysuccinate dissolved in 10 ml of tetrahydrofuran-dimethylsulfoxide (1: 1) in the cooled solution. A cooling solution such as imitate is added over 1 hour. After the resulting mixture was left for 2 hours at 0 ° C. and 1 night at room temperature, the mixture was treated in the same manner as described in Examples 1 (ii) and (iii) (A) to give 740 mg (70%) of a colorless solid. This compound is obtained.

+63°(c1, 물-디메틸포름아미드=1 : 2)

+ 63 ° (c1, water-dimethylformamide = 1: 2)

Elemental Analysis C ₄₂ H ₅₅ N ₅ O ₁₆ · 2CH ₃ CO ₂ HH ₂ O

Theoretical: C, 53.95; H, 6. 40; N, 6.84%

Anal: C, 53.66; H, 6.67; N, 6.63%

Treatment of this compound in a manner similar to that described in Example 31 yields 1-N-((S) -4-amino-2-hydroxybutyryl) ganamycin.

Example 20

Preparation of 3,2 ', 6'-tri-N-benzyloxycarbonyltobramycin

480 mg (1.03 mmol) of tobramycin (free base) is suspended in 12 ml of dimethylsulfoxide, 1 g (4.55 mmol) of zinc acetate dihydrate is added to the suspension, and the mixture is stirred at room temperature for 1 hour. After making a uniform solution, 850 mg (3.4 mmol) of N-benzyloxycarbonyloxysuccinimide solution dissolved in 10 ml of tetrahydrofuran-dimethylsulfoxide (1: 1) was added over 1 hour. When left at room temperature for one night, the reaction solution was treated with a large amount of ethyl ether as described in Example 1 (ii) to obtain a concentrated syrupy substance.

The syrup-like material was further treated in the same manner as in Example 1 (iii) (A) except that water-dioxane (1: 2 instead of 2: 1) was used to give 810 mg (78%) of a colorless solid. Obtain the compound.

+65°(c1, 물-디메틸포름아미드=1 : 2)

+ 65 ° (c1, water-dimethylformamide = 1: 2)

Elemental Analysis C ₄₂ H ₅₅ N ₅ O ₁₆ · 2CH ₃ CO ₂ HH ₂ O

Theoretic value: C, 54.81; H, 6. 50; N, 6.95%

Anal: C, 54.77; H, 6.71; N, 6.88%

Further treatment of this compound in a similar manner as described in Example 31 yields 1-N- (S) -4-amino-2-hydroxybutyryl) -tobramycin.

Example 21

Preparation of 3,2 ', 6'-tri-N-benzyloxycarbonyl-6'-methyltobramycin

The same procedure as in Example 20 was repeated, but using 500 mg (1.04 mmol) of 6'-N-methyltobramycin (free base) as a starting material, the compound was obtained in a yield of 890 mg (84%) as a colorless solid. Get

+63°(c1, 물-디메틸포름아미드=1 : 2)

+ 63 ° (c1, water-dimethylformamide = 1: 2)

Example 22

Preparation of 3,2 ', 6'-tri-N-benzyloxycarbonyl-4'-deoxyganamycin B

Example 20 using 480 mg (1.03 mmol) of 4'-deoxyganamycin B (freebase) (see "Bulletin of the Chemical Society of Japan Vol. 50, pp2362-2368 (1977)) as a starting material. In the same manner as in, the present compound in colorless solid form is obtained in a yield of 815 mg (79%).

+63°(c1, 물-디메틸포름아미드=1 : 2)

+ 63 ° (c1, water-dimethylformamide = 1: 2)

Example 23

Preparation of 3,2'-6'-triN-benzyloxycarbonyldibecasin

600 mg (1.33 mmol) dibecasin (3,4'-dideoxyganamycin B) (free base) are suspended in 15 ml of dimethylsulfoxide, the suspension is stirred and then 1.4 g (6.4 mmol). Acetic acid dihydrate was added, followed by further stirring. To the resulting solution was slowly added 1.1 g (4.4 mmol) of N-benzyloxycarbonyloxysuccinimide solution dissolved in 12 ml of dimethyl sulfoxide over about 1 hour, and the mixture was left at room temperature overnight. A large amount of ethyl ether is slowly added to the imide solution over about 1 hour, and then the mixture is left alone at room temperature. An oily precipitate obtained by mixing a large amount of ethyl ether with the reaction solution (a desired product and an amount of dimethyl sulfoxide consisting of N-benzyloxycarbonylated dibecacin-zinc complex) was separated and washed with ethyl ether. Get syrup substance.

The syrup-like material is washed repeatedly with water to decompose the N-acylated zinc complex with water to remove the free zinc cations with excess zinc acetate. 1.1 g of water-insoluble solid consisting of N-acylated dibecacin are obtained.

This solid was used as chloroform-methanol-18% ammonia water (1: 1: 1, lower layer) as a developing solvent to give a single point at Rf0.3 by silica gel thin layer chromatography. ', 6'-tri-N-benzyloxycarbonyldibecacin is obtained.

For further inspection, the crude product obtained as the title compound is washed with 3N ammonia solution to obtain a product free of zinc ion contamination.

+71°(c1, 물-디메틸포름아미드=1 : 2)

+ 71 ° (c1, water-dimethylformamide = 1: 2)

Example 24

Preparation of 3,2'-6'-tri-N-benzyloxycarbonyl-6'-N-methyldibecasin

500 mg (1.07 mmol) of 6'-N-methyldibecasin (free base) and 1.2 g (5.4 mmol) of zinc acetate dihydrate were dissolved in 20 ml of dimethyl sulfoxide, and 910 mg (3.6 m) in this solution. Mole) N-benzyloxycarbonyloxysuccinimide is added over 30 minutes. The reaction solution was left at room temperature for 1 night and then treated in the same manner as described in Example 23 to obtain pure 910 mg of the present compound.

Treatment of this compound in a manner analogous to that described in Example 31 yields 1-N-((S) -4-amino-2-hydroxybutyryl) -6'-N-methyldibecacin.

Example 25

Preparation of 3,2'-di-N-benzyloxycarbonylganamycin C

730 mg of the present compound, which is colored as a starting material, using the same method as described in Examples 1 (i), (ii) and (iii) (A), but using 500 mg (1.03 mmol) of kanamycin (free base) as a starting material. Obtained in a yield of (79%).

+75°(c1, 물-디메틸포름아미드=1 : 2)

+ 75 ° (c1, water-dimethylformamide = 1: 2)

Example 26

Preparation of 6'-N-benzyloxycarbonyl kanamycin A

500 mg (1.03 mmol) kanamycin A (freebase) are suspended in 20 ml dimethylsulfoxide, 0.5 g (2.3 mmol) zinc acetate dihydrate is added to the suspension and stirred at room temperature until a homogeneous solution is obtained. Then, 283 mg (1.13 mmol) N-benzyloxycarbonyloxysuccinimide is added. The resulting mixture was left at room temperature for one night, and then treated in the same manner as in Examples 1 (ii) and (iii) (I) to give 556 mg of the present compound as a colorless solid.

+92°(c1, 물)

+ 92 ° (c1, water)

Example 27

Preparation of 6'-N-benzyloxycarbonyldibecasin

In the same manner as described in Example 26, 500 mg of dibecasin (freebase) was dissolved in 12 ml of dimethylsulfoxide, and 0.7 g of zinc acetate dihydrate and 305 mg of N-benzyloxycarbonyloxysuccinimide Add 382 mg of the present compound.

+105°(c0.5, 물 )

+ 105 ° (c0.5, water)

Example 28

Preparation of 3,2'-6'-tri-N-benzyloxycarbonyl-3 ', 4'-dideoxy-3'-enoganamycin B

500 mg (1.11 mmol) of 3 ', 4'-dideoxy-3'-enoganamycin (free base) (see "Bulletin of the Chemical Society of Japan" Vol. 50, pp 1580-1583 (1977)) After dissolving in 12 ml of dimethyl sulfoxide, 1 g (4.55 mmol) of zinc acetate dihydrate was added thereto and stirred for 1 hour. 870 mg (3.40 mmol) of N-benzyloxycarbonyloxysuccinimide was slowly added to the resulting mixture over 30 minutes, and the reaction solution obtained after stirring at room temperature for 1 night was then described in Example 1 (ii). Treatment with large amounts of ethyl ether yields a syrupy material.

Treatment of the syrup-like material in the same manner as in Example 1 (iii) (B) afforded 784 mg of the present compound as a colorless solid using 1: 2 instead of water-dioxane 2: 1 (c1,) -Dimethylformamide = 1: 2).

Example 29

Preparation of 3,2 ', 6'-tri-N-benzyloxycarbonylsisomicin

Treatment with the method of Example 28 using 500 mg (1.12 mmol) of sisomycin (free base) as a starting material yields 780 mg of the present compound as a colorless solid.

+110°(c1, 물-디메틸포름아미드=1 : 2)

+ 110 ° (c1, water-dimethylformamide = 1: 2)

Example 30

Preparation of 3,2 ', 6'-tri-N-benzyloxycarbonylbentamycin

Treatment with the method of Example 28 using 500 me of mixed gentamicin (C, C ₁ a, C _2, etc.) as starting material yields 787 mg of this compound in the form of a colorless solid.

Example 31 (Comparative)

Preparation of 1-N-((S) -4-amino-2-hydroxybutyryl) kanamycin A (amicasin)

55 mg (0.062 mmol) of 3,6'-di-N-benzyloxycarbonyl kanamycin A acetate salt prepared as described in Example 1 were dissolved in 1.5 ml of water-tetrahydrofuran (2: 5). To this solution was added 13 mg (0.12 mmol) of anhydrous sodium carbonate, and then 23 mg (0.066 mmol) of (S) -4-benzyloxycarbonylamino-2-hydroxybutyric acid N-hydroxysuccinimide Ester is added. The mixture was allowed to stand at room temperature for 10 hours, the reaction solution was concentrated, the concentrate was dissolved in 4 ml of water-dioxane (1: 1), and a small amount of acetic acid was added to make the solution weakly acidic. And reduced to atmospheric hydrogen for 1 hour. (To remove benzyloxycarbonyl). The resulting reaction solution was concentrated by filtration, and the concentrate was added to a column of CM-Sepadex C-25 (NH ₄₄ form) (product of Pharmacia Fine Chemicals, Sweden), and the column was inclined to 0-0.5N ammonia water. Deploy it. Eluent fractions containing the desired product are collected together and concentrated to dryness to yield 24 mg (60% yield) of this compound as its monocarbonate, its physical properties and antibiotic potency being the same as for the certified sample.

Example 32 (comparative)

Preparation of 1-N- (DL-isoserine) dibecasin

58 mg (0.06 mmol) of 3,2 ', 6'-tri-N-benzyloxycarbonyldibecasin as prepared in Example 23 were dissolved in 1.5 m of water-tetrahydrofuran (2: 5) To this solution, 13 mg (0.12 mmol) of anhydrous sodium carbonate and 21 mg (0.063 mmol) of N-zylbenoxycarbonyl-DL-isoserine N-hydroxysuccinimide ester are added. The mixture was left at room temperature and treated in the same manner as described in Example 31 to obtain 21 mg of their monocarbonate (yield 59%) of the present compound, the physical properties and the antibiotic efficacy of which were the same as those of the certified sample. .

Claims (1)

The zinc complex of the aminoglycoside antibiotic produced by acting a zinc cation on the aminoglycoside antibiotic consisting of deoxystereptamine having a 3-aminoglycosyl group or a 3-alkylaminoglycosyl group at the 6-position is an amino-protecting group. By reacting with an acylating agent having an acyl group introduced into the compound, a zinc complex of an N-acylated derivative of an aminoglycoside antibiotic that is complexed with a zinc cation is produced, and zinc is derived from the zinc complex of the N-acylated derivative. A method for producing an N-protecting derivative of an ordinary American acemiglycoside antibiotic protected by an acyl group, characterized in that it is prepared by removing a cation.