CS207607B2 - Method of making the 1-n-/omega-amino-alphe-hydroxyalcanoylcanamycine a and b - Google Patents

Abstract

1-N-[ omega -amino- alpha -hydroxyalkanoyl] kanamycins A and B of the formula I <IMAGE> in which R represents OH or NH2, are prepared by acylation of polysilylated kanamycin A or B, or polysilylated kanamycin A or B which has a protective group other than silyl on the 6'-amino group, with an acylating derivative of an acid of the formula II <IMAGE> in which n is an integer from 0 to 2 and B is an amino protective group, in an essentially anhydrous organic solvent and by subsequent removal of all blocking groups. The compounds have therapeutic properties.

Description

(54) A method for producing 1-N- (n-amino-α-hydroxyalkanoyl) kanamycin A and B

The present invention relates to a process for the preparation of 1-způsobu- (ω-amino-α-hydroxyalkanoyl) kanamycin A B of the formula I

where

R is hydroxy or amino and n is an integer from 0 to 2 as well as non-toxic pharmaceutically acceptable acid addition salts thereof, characterized in that the polysilylated kanamycin A or B or the polysilylated kanamycin A or B is acylated with different from the silyl group on the 6'-amino group, provided that the polysilylated kanamycin A or B used as the starting material contains an average of 4 to 8 silyl groups per molecule, and the polysilylated kanamycin A or B used as the starting material and containing a 6-amino group protecting group other than a silyl group containing an average of 207607 the number of silyl groups in the 3 to 7 molecule, using an acid derivative of the formula II as acylating agent

В \

_B-N-CH2 - _{(CH2) n} -CH-COOH

OH (П) where n is an integer from 0 to 2, and

V is an amino protecting group or a hydrogen atom, provided that at least one of these symbols is an amino protecting group and then removes all protecting groups.

Kanamycins are well known antibiotics and have been described, for example, in the Merck Index, 8th Edition, pp. 597-598. Numerous kanamycin derivatives are also known. The structural formulas of kanamycin A and V are listed below along with the conventional numbering method for these formulas. Hereinafter, various kanamycin A or V derivatives will be referred to by name only and not by structural formula.

Kanamycins А and В can be represented by formula III

Kanamycin A: R = OH

Kanamycin B: R = NH 2

U.S. Pat. No. 3,781,268 describes a process for the preparation of 1- [L- (-) -? -Amino-.alpha.-hydroxybutyryl] kanamycin A (amikacin) and their protected derivatives using mono- and di-benzoyloxycarbonyl groups. Lower and higher homologs are described in U.S. Patent Nos. 3,886,139 and 3,904,597. The compounds are prepared by acylating kanamycin A or B protected at the 6'-amino group with an acylated derivative of N-protected L- (-) -y-amino-α- hydroxybutyric acid in aqueous medium followed by removal of one or both protecting groups.

U.S. Patent No. 3,974,137 discloses a process for the preparation of 1- [L- (-) - γ-amino-α-hydroxy-. butyryl] kanamycin A, which comprises reacting 6'-benzyloxycarbonylkanamycin A with at least 3 moles of benzaldehyde, substituted benzaldehyde or pivalaldehyde to give 6'-N-benzyloxycarbonylkanamycin A which contains the residue of the Schiff base at the 1,3-positions and 3 ', this kanamycin A derivative is acylated with L- (-) - γ-benzyloxycarbonylamino-α-hydroxybutyric acid N-hydroxysuccinimide ester and deprotected.

Belgian Patent No. 828,192 discloses a process for the preparation of 1- [L - (- Jy-amino-α-hydroxybutyryl) kanamycin A by preparing the same protected kanamycin A derivative as in US Patent No. 3,947,137 and acylating the derivative L- (-) -. Beta.-benzyloxycarbonylamino-.alpha.-hydroxybutyric acid N-hydroxy-5-norbornene-2,3-dicarboxyimide ester followed by deprotection.

The invention relates to an improved process for the preparation of compounds of the formula I. The use of polysilylated kanamycins A or V as starting materials ensures high solubility in the organic solvent and thus the reaction at high concentrations. Although the reaction is usually carried out in solutions containing 10 to 20% polysilylated kanamycin, very good results can be obtained at concentrations up to 50 g per 100 ml of solvent. *

As with known processes, a mixture of acylated products is obtained in the process of the invention. The desired 1-N-acylated product is separated from the other products by chromatography and, if desired, the by-products can be hydrolyzed to the starting kanamycin. In the known processes, it has been found that any amount of 3'-N-acylated material as the final product corresponds to the loss of approximately the same amount of the desired 1-N-acylated product, since the two products are very difficult to separate. A particularly advantageous feature of the process according to the invention is the formation of very low amounts of undesired 3 &apos; -N5

207807

usually not detectable at all.

The production of 1- [L- (-) -. Alpha.-amino-.alpha.-hydroxybutyryl] -canamycin A, which is also called amikacin, always results in a 3 &apos; -N-acylated product (BB-K11), a 3-N-acylated product (BB). -K29), 6'-N-acylated product (BB-K6), polyacylated material and, in addition, unreacted kanamycin A remains. This means that in the conventional manufacture of amikacin by acylation of 6'-N-benzyloxycarbonylcanamycin

And in an aqueous environment with subsequent deprotection, usually 10% of this substance, typically 2.5 kg per 25 kg, is lost due to the presence of BB-K11 as a by-product. When prepared according to the invention, the above is not possible. by-product in the reaction mixture at all.

Accordingly, the present invention provides a process for the preparation of 1-N- [ω-amino-α-hydroxyalkanoyl] kanamycin A and B 'of formula I

where

R is -OH or amino, and n is an integer from 0 to 2 as well as nontoxic. pharmaceutically acceptable addition salts of these. acidic substances, characterized in that:. The method according to claim 1, wherein the polysilylated kanamycin A or B or the polysilylated kanamycin A or B containing a different protecting group from the silyl group on the 6'-amino group is acylated, provided that the polysilylated kanamycin A or B used as starting material contains an average of 4 to 8 and the polysilylated kanamycin A or B used as starting material and containing a 6'-amino protecting group other than the silyl group contains an average number of silyl groups of 3 to 7, using an acid derivative of the general acid as the acylating agent. of formula II

B \

B - N - CH, - (CH 2) n - CH - COOH, I

OH (Π) where n represents an integer from 0 to 2,

B is an amino protecting group. or a hydrogen atom, provided that at least one of these symbols is an amino protecting group, namely o-nitrophenylthlos, a group of formulas

o II.

CM ОС - CH-C-OCHt I 3

AND

O Q

II II c- / YJCCC-,.

or the symbols V together form a group

where

R ¹ and R ² , the same or different, represent a hydrogen, fluorine, chlorine, bromine atom, a -NO 2, -OH, alkyl or alkoxy group having from 1 to 6 carbon atoms,

X represents a chlorine, bromine, fluorine or iodine atom, and

Y represents a hydrogen, chlorine, bromine, fluorine or iodine atom in a substantially anhydrous organic solvent followed by removal of all protecting groups, whereupon the resulting product is optionally converted to its non-toxic, pharmaceutically acceptable acid addition salt.

The acid of formula (II) may be used in its (+) or (-) form, or a mixture of both isomers, to give the corresponding compounds of formula (I) wherein the 1'-N-amino-α-hydroxyalkanoyl] group is also present in the in the form of (+) or (R) or in the form of (-) or (S), or a mixture of both. The individual forms and mixtures thereof are within the scope of the invention. According to a preferred embodiment, the acid of the formula II is used in the form of (-). In another preferred embodiment, the acid of formula II is used in the form of (+).

According to another embodiment of the process of the invention, a polysilylated kanamycin A or B, preferably polysilylated kanamycin A is used as the starting material. In another embodiment, a polysilylated kanamycin A or B, preferably polysilylated kanamycin A is used which contains a 6'-amino protecting group, other than the silyl group.

The most preferred protecting group is benzyloxycarbonyl.

In a preferred embodiment of the process according to the invention, the acylating agent is an acid of the formula II in the form of an active ester, preferably the active ester is N-hydroxysuccinimide ester, N-hydroxy-5-norfornene-2,3-dicarboximide ester and N-hydroxyphthalimide ester. According to a further preferred embodiment of the process according to the invention, the acid derivative of the formula II is a mixed anhydride, preferably with pivalic acid, benzoic acid, isobutylcarboxylic acid or benzylcarboxylic acid.

According to a highly preferred embodiment of the process of the present invention, N- [L- (-) - χ-amino-α-hydroxybutyryl] kanamycin A or a non-toxic pharmaceutically acceptable acid addition salt thereof can be obtained by acylating the polysilylated kanamycin A with mixed L- (- N -benzyloxycarbonylamino-α-hydroxybutyric acid anhydride, preferably a mixed anhydride with pivalic, benzoic, isobutylcarboxylic or benzylcarboxylic acid in a substantially anhydrous organic solvent, followed by removal of all protecting groups.

According to another highly preferred embodiment of the process of the invention, 1-N- [L- (-) -ylamino-α-hydroxybutyryl] cyanamycin A or a non-toxic pharmaceutically acceptable acid addition salt thereof can be obtained by acylating the polysilylated kanamycin A which contains a benzyloxycarbonyl group on the 6'-amino group with mixed L- (-) - γ-benzyloxycarbonylamino-α-hydroxybutyric anhydride, preferably a mixed anhydride with pivalic, benzoic, isobutylcarboxylic or benzylcarboxylic acid in a substantially anhydrous organic solvent followed by removal of all protective groups.

According to another highly preferred embodiment of the process of the invention, 1-N- [L- (-) -ylamino-3-hydroxybutyryl] kanamycin A or a non-toxic pharmaceutically acceptable acid addition salt thereof can be obtained by acylating the polysilylated kanamycin A active ester of L-

- (-) -? -Benzyloxycarbonylamino -? - hydroxybutyric, preferably N-hydroxysuccinimide ester, N-hydroxy-5-norbornene-2,3-dicarboximide ester or N-hydroxyphthalimide ester in a substantially anhydrous organic solvent followed by removal of all protecting groups .

In another highly preferred embodiment of the process of the invention, 1-N- can be obtained.

- [L- (-) - 4-Amino-α-hydroxybutyryl] kanamycin A or a non-toxic pharmaceutically acceptable acid addition salt thereof by acylating a polysilylated kanamycin A containing a benzyloxycarbonyl group at the 6'-amino group active an L- (N-4-benzyloxycarbonylamino-α-hydroxybutyrate) ester, preferably N-hydroxysuccinimide ester, N-hydroxy-5-norbornene-2,3-dicarboximide ester or N-hydroxyphthalimide ester in a substantially anhydrous organic solvent with

0 760'7 by sequential removal of all protecting groups.

Polysilylated kanamycin A or B containing different protecting groups from silyl groups on the 6‘-amino group is used as starting material in the process of the invention. According to a preferred embodiment of the process according to the invention, the material is polysilylated kanamycin A or B, preferably polysilylated kanamycin A, containing on average 4 to 8 silyl groups, preferably trimethylsilyl groups per molecule. According to another preferred embodiment of the process according to the invention, the material is polysilylated kanamycin A or B, preferably polysilylated kanamycin A, which contains a 6'-amino protecting group different from the silyl group and otherwise contains 3 to 7 silyl groups, preferably trimethylsilyl groups groups in the molecule.

The term "nontoxic, pharmaceutically acceptable acid addition salt" of a compound of formula (I) refers to a quadruple salt of a compound of formula (I), obtained by reaction with 1 to 4 equivalents of a non-toxic, pharmaceutically acceptable acid. These include, for example, acetic, hydrochloric, sulfuric, maleic, phosphoric, nitric, hydrobromic, ascorbic, malic and citric acids and other acids commonly used for this purpose.

The acylation of the starting polystilylated kanamycin A or B, which optionally contains a 6‘-amino protecting group other than the silyl group, is usually carried out in an organic solvent in which the starting material is sufficiently soluble. The starting materials are highly soluble in most organic solvents. Suitable solvents are, for example, acetone, diethyl ketone, methyl-n-propyl ketone, methyl isobutyl ketone, methyl ethyl ketone, acetonitrile, glyme, diglyme, dioxane, toluene, tetrahydrofuran, cyclohexanone, methylene chloride, chloroform, carbon tetrachloride and mixtures of acetone with butanol or diethyl ketone with butanol. The choice of solvent depends on the starting materials used. Ketones are the most suitable solvents. The most preferred solvent for a particular mixture of reactants is readily determined by simple preliminary steps.

Suitable silylating agents for the examples of starting polysilylated kanamycins are, for example, compounds of formula IV and V

H (IV)

R ⁵

R6 — Si — X,

R7

(V) where

R ⁵ , R ^6, and R 7 represent a hydrogen atom, a halogen atom, a lower alkyl group, a lower haloalkyl group or a phenyl group, at least one of which has a different meaning from a halogen atom or a hydrogen atom,

R4 represents lower alkyl, m represents 1 to 2 and

X represents a halogen atom or a group

R ⁹ where

R 8 is hydrogen or lower alkyl, and

R ⁹ represents a hydrogen atom, a lower alkyl or a group

R ⁵

R6 —SIL—,. R7 where

R ⁵ , R ® and R 7 are as defined above.

Specific silylating agents of formulas IV and V are the following compounds:

trimethylchlorosilane, hexamethyldisilazane, triethylchlorosilane, methyltrichlorosilane, dimethyldichlorosilane, triethylbromosilane, tri-n-propylchlorosilane, methyldiethylchlorosilane, dimethylethylchlorosilane, dimethyl t-butylchlorosilane, phenyldimethylbromosilane,

7 6.0 7 benzylmethylethylchlorosilane, phenylethylmethylchlorosilane, triphenylchlorosilane, triphenylfluorosilane, tri-o-tolylchlorosilane, tri-p-dimethylaminophenylchlorosilane, N-ethyltriethylsilylamine, hexaetbyldisllazane, triphenylsilylisilyl, tri- n -silylisilane, tri- n -silylisilane, tri-n-ethylsilane;

Other useful materials are hexaalkylcyclotrisilazanes and octaalkylcyclotetrasilazanes. Useful silylating agents are also silylamides, such as trialkylsilylacetamides and bis-trialkylsilylacetamides, silylureas, such as trimethylsilylurea and silylureides. Trimethylsilylimidazole may also be used.

A preferred silyl group is a trimethylsilyl group, and a preferred silylating agent for the introduction of this group is hexamethyldisilazane, bis (trimethylsilyl) acetamide and trimethylsilylacetamide. Most preferred is hexamethyldisilazane.

When polysilylated kanamycin is used as the starting material. A or B having a protecting group on the 6'-amino group different from the silyl group can be obtained either by introducing several silyl groups into the kanamycin A or B molecule already protected at the 6'-amino group or the desired protecting group is introduced into the 6 'position in polysilylated kanamycin A or B.

Methods for introducing silyl groups into organic compounds including some. aminoglycosides are known. Polysilylated kanamycins, even those containing a 6‘-amino protecting group other than a silyl group, can be made by known methods or by methods described during the application.

The term "polysilylated kanamycin A or B" refers to kanamycin A or B containing 2 to 10 silyl groups per molecule. That is, the term does not include persilylated kanamycin A or B containing 11 silyl groups. in the molecule.

The exact number of silyl groups and their position in the starting polysilylated kanamycins which optionally contain a 6‘-amino protecting group different from the silyl group is not known. Too little or too many silyl groups have been found to reduce the yield of the desired product and increase the yield of by-products. If the number of silyl groups is really small or indeed too large, it may happen that only a very small amount of the desired group is formed. product or this product does not arise at all. The degree of silylation at which the highest yield of the desired product can be obtained depends on the specific reactants in the acylation stages. The most preferred degree of silylation can be determined by simple preliminary experiments for each reactant.

In the case where 1-N- [L- (-) -? -Amino-.alpha.-hydroxybutyryl] kanamycin A is prepared by acylation of polysilylated kanamycin A with N-hydroxysuccinimide ester of L- (-) -? -Benzyloxycarbonylamino-.alpha.-hydroxybutyric acid in acetone. · Solution, it has been found that it is possible to achieve good yields of the desired product when using polysilylated · kanamycin · A, prepared by reaction 4 to 4

5.5 moles of hexamethyldisilazane per mole of kanamycin A. Although higher or lower amounts of hexamethyldisilazane may be used, the yield of the desired product in the subsequent acylation step is substantially reduced. Most preferred is 4.5. up to 5.0 moles of hexamethyldisilazane per 1 mol of kanamycin because of this. in this case, the maximum yield of the product can be achieved in the acylation step.

It is understood that each mole of hexamethyldisilazane can be introduced. to · kanamycin A or B · 2 equivalents of the trimethylsilyl group. Kanamycin A and B have a total of 11 positions (NHa and OH groups) which can be silylated, while kanamycin A and B which contain a 6'-amino protecting group different from the silyl group This means that, when 5.5 mole of hexamethyldisilazane is used per mole of kanamycin A or B, it is theoretically possible to silylate all of the hydroxy and amino groups of kanamycin, while at 5.0. moles of hexamethyldisilazane can be completely subjected to silylation of 1 mol of kanamycin A. or B containing 6‘-amino. a protective group, other than a silyl group. However, it appears that using the said molar amounts. there is no complete silylation if the reaction time is not too long, although it must be recognized that a high degree of silylation can be achieved in the shorter time.

Silylation catalysts greatly increase its velocity. Suitable catalysts. silylations that are generally well known are, for example, amine sulfates such as kanamycin sulfate, sulfonic acid, imidazole and trimethylchlorosilane. Silylation catalysts usually produce a higher degree of silylation than. Is required during the process of the invention. Kanamycin A or B having a large number of silylated groups can be used as starting materials in the process of the invention when subjected to a desilylating agent to reduce the degree of silylation prior to the acylation reaction.

Good yields of the resulting product can be obtained when acylating the polysilylated kanamycin A prepared using a ratio of hexamethyldisilazane to kanamycin A of 5.5: 1. When acy207607 is used for kanamycin A silylated at a 7: 1 hexamethyldisilazane ratio, or · At a ratio of 5.5: 1 when using a silylation catalyst in an acetone solution with N-hydroxysuccinimide ester of L- (-) -? -Benzyloxycarbonylamino-α-hydroxybutyric acid, less than 1% of the desired product is obtained . When the same kanamycin A is acylated with the same acylating agent in an acetone solution to which water was added at a rate of 21 moles of water per 1 mol of kanamycin, which means 2.5% water (w / v) 1 hour before acylation as about 40% of the desired end product. The same results were obtained when methanol or other substances capable of desilylating such as ethanol, propanol, butanediol, methylmercaptan, ethylmercaptan, phenylmercaptan and the like were used instead of water.

Although it is customary to use an anhydrous solvent to work with silylated materials, it has now unexpectedly been found that even when the material used does not contain too many silyl groups, the addition of water to the reaction solvent prior to acylation yields equally good and sometimes better results than using an anhydrous solvent. In acylation reactions carried out in acetone at a conventional concentration of 10 to 20% (w / v ·%) of polysilylated kanamycin A, it was found that very good yields of 1-N- [L- (-) -? -? [alpha] -amino-.alpha.-hydroxybutyryl] kanamycin A when up to 28 moles are added per mole of polysilylated kanamycin A. At a concentration of 20%, 28 moles of water is approximately 8% water. If another mixture of reactants is used, it is sometimes possible to add even more water. The acylation reaction can be carried out in solvents containing up to 40% water, although at such a high water concentration the acylation time must be shortened in order to avoid too much desilylation of the polysilylated kanamycin A or B. It means that the term · "Essentially anhydrous organic solvent" is intended to include solvents containing up to 25% water. The preferred range is up to 20% water, even more preferably up to 8% water and most preferably up to 4% water.

As mentioned above, the most preferred degree of silylation for individual reactant mixtures during acylation can be readily determined by conventional experiments. The preferred average amount of silyl groups in the starting material is usually in the range of 4 to 8 for kanamycin A or B and 3 to 7 for kanamycin A or B, which contains a 6'-amino protecting group different from the silyl group but only theoretical considerations that do not form a substantial part of the invention.

The temperature and acylation time are not critical values. The temperature may range from -30 to 100 ° C and the reaction time may range from 1 hour to 1 or more days. It has been found that the reaction usually proceeds well at room temperature and was therefore carried out at this temperature. When maximum yields or selective acylation are desired, the acylation must be carried out at 0-5 ° C.

Acylation of 1-amino polysilylated kanamycin A or B, optionally containing ⁴ to 6? -Amino protecting group, different from the silyl group can be effected by any acylating derivative of an acid of formula II, which can be performed by acylation of the primary amino groups. Examples of suitable acylating derivatives of the free acid are anhydrides, mixed anhydrides, for example mixed anhydride with alkoxy formic acid, acid halides, its azides, active esters and active thioesters. The free acid may be reacted with the starting polysilylated kanamycin by first reacting the free acid with N, N ⁴ -dimethylchlorforminium chloride according to British Patent No. 1,008,170 or by Novák and Weichet, Experientia XXI, 6 , 360 (1965) or using N, N'-carbonyldiimidazole or N, N'-carbonylditriazole according to South African Application No. 62/2684 or a carbodiimide type reagent, especially N, N'-dicyclohexylcarbodiimide, N, N diisopropylcarbodiimide or N-cyclohexy-N '- [2-morpholinethyl) carbodiimide according to Sheehan and Hess, JACS, 77, 1967 (1955), alkynylamine [R. · Buijle and HG Viege, Angew. Chem. International Edition, 3, 582 (1.96-4 ·)] or isoxazolium salts [RB Woodward, RA Olofson and H. Mayer, J. Amer. Chem. Soc., 83, 1010 (1961)] or ketenimine [CL Stevens et al. E. Munk, J. Amer. Giem. Soc., 80, 4065 (1958)] or hexachlorocyclotrifosphatriazine or hexabromocyclotrifosphatriazine according to U.S. Patent No. 3,610,505, diphenylphosphoryl azide [DDPA, J. Amer. Chem. Soc., 94, 6203-6205 (1972)], diethylphosphoryl cyanide [DEPC, Tetrahedron Letters No. 18, pp. 1595-1593 (1973)] or diphenylphosphite [Tetrahedron Letters No. 49, pp. 5047-5050 19 * ^ 22)]. Another equivalent of the acid is the corresponding azolid, ie the amide of the corresponding acid in which the nitrogen atom of the amide group is simultaneously a member of a five-membered ring containing at least 2 nitrogen atoms such as imidazole, pyrazole, triazoles, benzimidazole, benzotriazole and substituted derivatives of these substances. It will be appreciated that sometimes it may be desirable or necessary to protect the hydroxyl group of the acylating derivative of the acid of formula II, for example, when an acid halide is used as the acylating derivative. Protection of the hydroxyl group can be ensured in a known manner, for example using benzyloxycarbonyl, acetylation, silylation and the like.

Upon completion of the acylation reaction, all of the protecting groups are removed in a known manner to give the desired product of formula (I). For example, silyl groups can be readily removed by hydrolysis in water, preferably at a lower level. pH. The protecting group V of the acid acylation derivative of the formula II and the protecting group on the 6‘-amino group of the polysilylated starting kanamycin, if present, can also be removed by known methods. This means that the tert-butoxycarbonyl group can be removed using formic acid, the benzyloxycarbonyl group by catalytic hydrogenation, the 2-hydroxy-1-naphthylcarbonyl group can be removed by acid hydrolysis, the trichloroethoxycarbonyl group by treatment with zinc powder in glacial acetic acid, simultaneous heating, etc.

The yields of the resulting products can be determined in various ways. After removal of all protecting groups and chromatography on a column of CG-50 in NHt ⁺ form, the yield of amikacin can be determined by isolating the crystalline solid from the appropriate fractions or by microbiological detection of the presence of this substance by turbidimetry or on plates using appropriate fractions. Other methods may be used, for example, liquid chromatography of an unreduced acylation mixture, i.e., an aqueous solution obtained after hydrolysis of the silyl groups and removal of the organic solvent but prior to hydrogenolysis to remove the remaining protecting groups. This reaction is not a direct reaction to amikacin or BB-K29, it is a reaction to identify the corresponding mono- or di-N-protected compounds.

Waters Associates ALC / GPG 244 and Waters Associates Model 440 absorbance detector using a 30 cm x 3.9 mm µ-Bondapak C-18 column were used in these tests. under following conditions:

Mobile phase:

25% 2-propanol,

75% 0.01 M sodium acetate at pH 4.0. · »

Flow rate:

ml / min. Detector:

spectrometer for the determination of ultraviolet light at 254 nm.

Sensitivity: _ 0.04 AUFS.

Thinner:

dimethylsulfoxide.

Amount used:

μΐ.

Concentration:

mg / ml.

The recording speed was varied, but typically a rate of about 2.5 cm per 2 minutes was used. Under the above conditions, quantitative measurable peaks were obtained in ultraviolet light. The results of these analyzes will hereinafter be referred to as HLPC assays.

The following abbreviations will be used throughout the application to avoid the recurrence of complex chemical names:

AHBA - L- (-) -y-amino-N-hydroxybutyric acid

BHBA - N-benzyloxycarbonyl derivative AHBA HONB - N-hydroxy-5-norbornene-2,3-dicarboximide

NAe (or BHBA- “ONB” J - activated N-hydroxy-5-norbornene-2,3-dicarboximide ester BHBA

HONS-N-hydroxysuccinimide

SAE (or BHBA- "ONS") - activated N-hydroxysuccinimide ester BHBA DCC - dicyclohexylcarbodiimide DCU - dicyclohexylurea HMDS - hexamethyldisilazane BSA - bis (trimethylsilyl) acetamide MS A - trimethylsilylacetamide TFA - trifluoroacetyl t-BOC-tert-butylacetyl t-BOCx.

"Dicalit" is a diatomaceous earth product supplied by Great Lakes Carbon Corporation.

"Amberlit CG-50" is a weakly acidic cation exchange resin of the carboxylic acid and polymethacrylic acid type, supplied for chromatography by Rohm and Haas Co.

“μ-Bondapak” is a column packing supplied for high pressure liquid chromatography by Waters Associates.

The terms "lower alkyl" and "lower alkoxy" mean alkyl and alkoxy groups containing 1 to 6 carbon atoms.

The invention will be illustrated by the following examples in which all temperature data are given in degrees Celsius.

Example 1

Method for the preparation of N - [.alpha .- (. Beta.-amino-.alpha.-hydroxybutyryl) kanamycin A (amikacinine by selective acylation of poly (trimethylsilyl) -6'-N-benzyloxycarbonylkanamycin A in anhydrous diethyl ketone g (24.24 mol) 6'-N The benzyloxycarbonylcanamycin A is suspended in 90 ml of anhydrous acetonitrile and the suspension is heated to reflux under a nitrogen atmosphere, then 17.5 g (108.48 mol) of hexamethyldisilazane are slowly added over 30 minutes and the resulting solution The solvent was removed in vacuo at 40 ^° C and the resulting solid was dried at 50 psi to give 27.9 g of a white amorphous solid in a yield of 90.71 ° C. calculated for 6'-N-benzyloxycarbonylkanamycin A with 9 silyl groups.

The solid obtained is. Dissolve in 150 ml of anhydrous diethyl ketone at 23 ° C. Then, 11.05 g (26.67 mmol) of N-hydroxy-5-norbornene-2,3-dicarboximide ester of L- (γ-benzyloxycarbonylamino-α-hydroxybutyric acid) in solution in L-hydroxybutyric acid are slowly added over 30 minutes with vigorous stirring. 100 ml of anhydrous diethyl ketone at 23 DEG C. The solution is stirred at 23 DEG C. for 78 hours, and the yellow clear pH 7.0 is diluted with 100 ml of water and the pH of the mixture is adjusted to 2.8 by adding 3 N. hydrochloric acid is added, and the mixture is stirred vigorously for 15 minutes at 23 DEG C. The aqueous phase is separated and the organic phase is extracted with 50 ml of water at pH 2.8 and the aqueous phases are combined, washed with 50 ml of ethyl acetate. Parr bottle of 500 ml containing 5 g of 5% palladium on charcoal, then reduced for 2 hours at 23 ° C at a pressure of 50 psi (50 psi) and filtered through a pad of Dicalit, which was then filtered. rinse with 30 ml of water, concentrate the colorless filtrate · The solution was then applied to a 5 x 100 cm column containing CG-50 in NH4 + form. The column is washed with 100 ml of water and then unreacted kanamycin A, 3- [L- (-] -. Alpha.-amino-.alpha.-oxybutyryl] kanamycin A (BB-K29) and the amikacin are eluted with 0.5 M ammonium hydroxide. It is isolated by the addition of 3 N ammonium hydroxide Biological assays, thin layer chromatography and optical rotation are used to demonstrate elution of the active material, the volume of each fraction, its rotation and the amount of the substance obtained, together with the yield after evaporation of each fraction to dryness. in the following table:

Material Volume (ml) kanamycin A1000

BB-K291750 amikacin2000 polyacylated derivatives900

^and 578 Weight (g) Yield (%) + 0.115 0,989 9.15 + 0.24 4.37 32.0 + 0.31 6.20 47.4 + 0.032 0.288 2,0

HPLC showed that the deposited layer of ethyl ketone still contained 3-5% amikacin.

Dissolve 6.20 g of crude amikacin in 20 ml of water and dilute the solution with 20 ml of methanol. 20 ml of isopropanol are added to start crystallization. 6.0 g of crystalline amikacin are obtained in a yield of 45.8%.

Example 2

A process for the preparation of 1-N- (L- (-) -y-amino-α-hydroxybutyryl] kanamycin A (amikacin) by selective acylation of (trimethylsilyl) -6s-N-benzyloxycarbonylkanamycin A in anhydrous acetone

103 g (0.081 mol) of poly (trimethylsilyl) -6'-N-benzyloxycarbonylkanamycin A prepared as described in Example 1, calculated on 6'-N-benzyloxycarbonylcanamycin A (silyl) g, are dissolved in 100 ml of anhydrous acetone at 23 ° C. . Then, 35.24 g (0.085 mol) of L- (-) -χ-benzyloxycarbonylamino-α-hydroxybutyric acid N-hydroxy-5-norbornene-2,3-dicarboxyimide ester in solution in 180 ml of anhydrous acetone is added at 23 ^{° C.} C slowly with vigorous stirring to a solution of (trimethylsilyl) -6'-N-benzyloxycarbonylcanamycin A poles over 15 minutes. The solution was stirred at 23 ° C for 20 hours under a nitrogen atmosphere. A pale yellow, clear solution of pH 7.2 is obtained, which is diluted with 100 ml of water. The mixture was adjusted to pH 2.5 by addition of 3 N hydrochloric acid and then stirred for an additional 15 minutes at 23 ° C. Acetone was removed at 35 ° C by distillation with water vapor under vacuum. The solution was then placed in a 500 ml Parr bottle along with 10 g of 5% palladium on activated carbon catalyst and the mixture was reduced at room temperature for 2 hours. at 23 ° C and at a hydrogen pressure of 0.28 MPa. The mixture is filtered through a pad of diatomaceous earth, which is then washed with a further 50 ml of water. After concentration to approximately one third of the initial volume, the solution is at pH. 6.9 to 7.2 are applied to the top of a 6 x 110 cm column containing CG-50 in NH4 ⁺ form, and the column is eluted with water with an &quot; increasing amount of ammonium hydroxide &quot; concentration · 0.6 N for amikacin elution. In order to monitor the elution of this substance,. use an automatic polarimeter. At the same time, evaluation by thin layer chromatography is performed. The fractions containing amikacin are pooled and concentrated to a concentration of 25-30%. The solution is diluted with an equal amount of methanol and then 2 volumes of isopropanol are added to induce crystallization. 18.2 g of crystalline amikacin are obtained in a yield of 40%.

12% kanamycin A, 40% BB-K29, and 5% polyacylated kanamycin were also isolated, yielding a total of 97% of the product.

Example 3

Process for the preparation of 1-N- [L- (-) -? -Amino-.alpha.-hydroxybutryl] kanamycin A (amikacin) by selective acylation of poly (trimethylsilyl) kanamycin A in introducing protecting groups directly in the reaction mixture

A. Kanamycin A (trimethylsilyl) poles (37.15 mmol) in free form are suspended in 200 ml of anhydrous acetonitrile and the suspension is heated to reflux. cooler. Hexamethyldisilazane (29.8 g, 184.6 mmol) was then added over 30 minutes and the resulting mixture was heated at reflux for 78 hours to give a clear pale yellow solution. The solvent was evaporated in vacuo to give 43 g of an amorphous solid, 94% yield, calculated as kanamycin A (silyl).

B. 1-N- [L- (-) -y-amino-.alpha.-hydroxybutyryl] kanamycin A

5.56 g (20.43 mmol) of p- (benzyloxycarbonyloxy) benzoic acid are mentioned in m.p. suspension in 50 ml of anhydrous acetonitrile at 23 ° C. 8.4 g [41.37 mmol] of N, O-bis-trimethylsilylacetamide are then added with vigorous stirring. The solution is maintained at 23 for 30 minutes. ° C and then added over 3 hours with vigorous stirring to a solution containing 21.5 g (17.83 mmol) of poly (trimethylsilyl) k-anamycin A, calculated on the compound with 10 silyl groups in 75 ml of anhydrous acetonitrile. at 23 ° C. The mixture was stirred for 4 hours, then the solvent was removed in vacuo. . vacuum at 40 ° C and the oily residue was dissolved in 50 ml of anhydrous acetone at temperature. Deň: 18 ° C.

8.55 g. (20.63 mmol) of N-hydroxy-5-norbornene-2,3-dicarboimidester. L- (-) - γ-benzyloxycarbonylamino-α-hydro-. xbuttered (NAE) ve. 30 · ml. acetone. is added to the above solution over a period of 5 minutes. The mixture was held for 78 hours. at 23 ° C. Then take a solution. Dilute with 100 ml. water and pH was reduced from 7.0 to 2.5 by addition of 6 N acid. hydrochloric acid. The mixture obtained. is placed in a Parr container. content. 500 ml simultaneously with 3 g of 5% palladium on activated carbon and then reduced by 2. hours at 23 cq. pressure. of hydrogen 0.28 MPa. . The mixture was then filtered through a pad of diatomaceous earth, which was further washed with 20 ml of water. The filtrate. and the wash water in a total amount of 168 ml is microbiological. tests using E. coli, which makes it possible. showed approximately 11 400 µg / ml amikacin, which means a yield of 19

Example 4

Process for the preparation of 1-N- [L- (-) -y-amino-.alpha.-hydroxybutyryl] kanamycin A (amikacin) by selective acylation of (trimethylsilyl) kanamycin A poles

Polymers (trimethylsilyl) kanamycin

A suspension of 10 g (20.6 mmol) of kanamycin

And in 100 ml of anhydrous. acetonitrile. and 25 ml (119 mmol) of 1,1,1,3,3,3-hexamethyldisilazane was heated at reflux for 72 hours. A pale yellow, clear solution is obtained which is distilled under vacuum to dryness. at 30 ^Q C. 40 .When this manner. 21.3 g of poly (trimethylsilyl) kanamycin A are obtained in the form of an off-white amorphous powder in a yield of 85% calculated as kanamycin A containing 10 silyl groups.

B. 1-N- [L- (-) -y-amino-.alpha.-hydroxybutyryl] kanamycin A

To a solution of 2.4 g (2.0 mmol) of poly (trimethylsilyl) cyanamycin A in 30 ml of anhydrous acetone was slowly added 2.0 mmol of L- (-) - N-hydroxy-5-norbornene-2,3-dicarboximidoester. The .alpha.-benzyloxycarbonylamino-.alpha.-hydroxybutyric acid (NaE) in 10 ml of anhydrous acetone at 0-5 [deg.] C. The reaction mixture was stirred at 23.5-0 for a week, then evaporated in vacuo. 60 ml of water and then 70 ml of methanol were added to the residue to obtain a solution, and the resulting solution was acidified to pH 2.0 with 3 N acid. hydrochloric acid and then reduced for 2 hours at a hydrogen pressure of 50 psi using 500 mg of 5% palladium on activated carbon as the catalyst. The material obtained is filtered, filtrates and a washing liquid. They were combined and subjected to microbiological tests using E. coli to demonstrate amikacin in a yield of 29.4%.

Exampleb

A process for the preparation of amikacin by selective N-acylation of polytrimethylsilyl-6‘-N-benzyloxycarbonylcanamycin A in anhydrous form. acetone

I. General description. . ... '... ·

Silylation. N, N-benzocarbonylcanamycin A in acetonitrile using hexamethyldisilazane (HMDS) affords 6'-N-benzoxycarbonylcanamycin A with 9 silyl groups such as. Intermediate I. This material is well soluble in most organic. solvents. Acylation by action. NAE in anhydrous acetone at 23 cq and a 5 molar excess of NAE relative to the starting kanamycin A derivative yielded a mixture containing only the benzoxycarbonyl (Cbz) amikacin derivative and BB-K29, unreacted kanamycin A and a small amount of polyacylated material. BB-K11 could not be detected. Elution of the acylation mixture after reduction and column treatment. CG-50 in NH4 ⁺ form * and using ascending. The amount of ammonium hydroxide is pure amikacin in 40% yield.

II.

AND.

+ (CH 3) 3 Si-NH-S 1 (CH 3) 3

HMDS (161.4)

B.

DCC (206) acetone

BHBA (253.4) HONB (179.2)

OH o

I II

DCU + (CU ^U £ CH ~ C ~ o

®NAE (414.6) (224.3)

С.

diCbzamtkacln (853)

acetone ------> <

^C C diCbzBB K29 +

/ ----------------- amikacin BB-K29 4 * '№

5% Pd / C / 4.

6‘-Cbz-1,3-diBHBA-kanamycin A +

6 ‘~ Cbz: kanamycln A к

1,3-diAlBa-kanamycin A + kanamycin A (585.62) (484.5)

arnik acin

III. Materials

Weight (g) Volume (ml) Mol

6‘-N-Cbz *) kanamycin A 50 0,081 HMDS 58.9 76.5 0.365 acetonitrile 300 BHBA 21.5 0,085 HONB 15.2 0,085 DCC 17.48 0,085 acetone 260 CG-50 (NH ⁺ ) 3000 methanol as required IPA as required

*) Cbz = benzoxycarbonyl

IV. Security precautions .

6'-N-Cbz kanamycin A - no direct information available, avoid contact with powdered acetonitrile - same procedure as cyanide, avoid inhalation of vapors, may cause skin irritation hexamethyldisilazane (HMDS) - all-round irritant , the need for careful treatment

6‘-N-Cbz kanamycin A (silyl) 9 - no information available, contact with powdered material should be avoided,

BHBA - toxicity not determined, contact with solid material should be avoided

HONB - toxicity is not known, common safety measures are sufficient

DCC - a substance that strongly irritates the skin and eyes. It is necessary to avoid inhalation of vapors, the substance It is very toxic acetone - it is flammable, after inhalation it shows headache, fatigue, later arousal, bronchial irritation and anesthesia in large quantities

NAE - information is not available, but methanol is always used in acetone solution - it is flammable, poisoning can be caused not only by ingestion but also by inhalation or absorption of the skin.

isopropanol - is flammable, after ingestion or inhalation of large amounts of vapors, headache, clogging, vomiting and anesthesia ammonium hydroxide - vapors are toxic, possible. work in a mask and avoid. in contact with liquid

CG-50 (NHd ⁺ j - toxicity data not available, normal precautions are sufficient.

In the embodiment

A. Method for the preparation of 6‘-N-eenzyloxycarbonylkanamycin A (silyl β 6-N-Cbz kanamycin A (silyl) 9j

1. 50 g of 6‘-N-benzyloxycarbonylcanamycin A (KF <4%) are suspended in 300 ml of acetonitrile (KF <0.01%). The mixture was brought to 74 ° C under reflux while an anhydrous nitrogen stream was passed through the slurry.

2. Add slowly over 30 minutes

75.8 ml hexkmethyldisilkzane (HMDS). This creates a clear one. solution while producing ammonia gas.

3. The mixture is refluxed for 18 to 20 hours under nitrogen. atmosphere.

4. Clear, light. the yellow solution is concentrated under reduced pressure at bath temperature 4. up to 50 ° C to a foamed solid. Gets. 89-92 g of a compound having 9 silyl groups, i.e. a yield of 90-94% of the theoretical.

Note: When other solvents are used, this solid is usually not isolated but used. directly to acylation.

B. Method for the preparation of L- (-) -?

1. 21.5 g of L- (-) - t -benzyloxycarbonyl-amino-1-hydroxybutyric acid (BHBA). Dissolve in 100 ml of anhydrous acetone at 23 ° C and then add 15,2 N-hydroxy-5-norbornene-2,3-di-crboximide (HONB).

A solution is formed.

2. Add a solution of 17.48 g of dicyto-oxoyl-carbodiimide (DCC) in 50 ml of acetone with stirring over 30 minutes. The temperature rises to approximately 40 ° C during the addition while precipitating dicyclohexylurea (DCU).

3. Stir the suspension for 3 to 4 hours while maintaining the temperature. on . 23 to 25 ° C.

4. The urea derivative is removed by filtration, the filter cake is washed with 30 ml of acetone. The filtrate and washings are discarded for the acylation step.

C. Acylation of 6‘-N-Cbz kanamycin A (silyl) g

1. Dissolve 6‘-N-Cbz kanamycin A (silyl 19 isolated in Part A, Step 4) in 100 ml of anhydrous acetone at a temperature of 23 to 24 ° C.

2. With vigorous stirring, slowly add the NAE solution prepared in Part B over 15 minutes. The temperature gradually rises to approximately 40 ° C. The temperature of the solution was then allowed to stabilize at 23 ° C and the mixture was further stirred for 18-20 hours under a nitrogen atmosphere.

3. Add 100 ml of water and lower the pH, which initially has a value of 6.9 to 7.2, to a value of 2.2 to 2.5 by adding 6 N hydrochloric acid. The mixture was stirred at 23 ° C for 15 minutes.

Note: A second layer may be formed, but this does not affect the further processing of the mixture.

4. Remove the acetone under reduced pressure at a bath temperature of 30 to 35 ° C. The concentrate is transferred to a suitable hydrogenation vessel which is pre-flushed with nitrogen. 10 g of 5% palladium-on-carbon catalyst are then added and the hydrogenation is carried out for 2 to 3 hours at 0.28 MPa.

5. Filter the mixture through a Dicalit layer, wash the hydrogenation vessel and wash the filter cake with another 50 mL of water.

6. Concentrate the filtrate and wash water to one third of the volume (50 mL) under vacuum at 40 to 45 ° C.

7. It is necessary to control the pH, which should be in the range 6.9 to 7.2. If the pH is lower, it is necessary to adjust by adding 1 N ammonium hydroxide. The mixture was then applied to a 6 x 110 cm column containing CG-50 (NH4 ⁺ ).

8. Wash the column with 1000 ml deionized water and then elute with 0.5 to 0.6 N hydroxide using an automatic polarimeter to monitor elution. The components are eluted in the following order:

the remaining kanamycin A,

BB-K29 and amikacin.

BB-K11 could not be detected in any acylation reaction mixture. The polyacylated material, i.e. the 1,3-diAHBA analog of kanamycin A, can be isolated by washing the column with 3 N ammonium hydroxide.

9. The amikacin fractions are combined and concentrated to contain 25-30% solids. The concentrate was then diluted with an equal volume of methanol and crystallization was initiated by the addition of ¹ small amikacin crystals.

10. Add 2 volumes of isopropanol slowly over 2 hours with vigorous stirring, then allow the mixture to crystallize for 6 to 8 hours at 23 ° C.

11. The solid is collected by filtration, washed with 50 ml of a 1: 1: 2 mixture of water, methanol and isopropanol and finally 25 ml of isopropanol.

12. The crystals are dried under vacuum at 40 ° C for 12-16 hours. This gives 17.3 to 19.0 g of amikacin in a yield of 38 to 42%. Amikacin has the following properties:

Thin Layer Chromatography • Using a 1: 4: 2: 1 mixture of chloroform, methanol, ammonium hydroxide and water and 5x20 cm silica gel plates (Quantum Industries), a single zone is formed as can be shown ninhydrin (R _f = 0.4).

Specific rotation

H 2 O 0.1 Μ NH 4 OH 0.1 M H 2 SO 4 (α) 589 ²³ + 101.6 + 101.9 + 103.5

C = 1.0 0/0.

13. The yield of BB-K29 in this system was also 39-42%, the mixture contained 10-14% kanamycin A and approximately 5% 1,3-di-AHBA kanamycin A, thus recovering approximately 95% of the original materials.

Example 1 a d 6

A process for the production of amikacin by selective acylation of polytrimethylsilyl-kanamycin A in anhydrous acetone

I. General description

Silylation of kanamycin A in free form in acetonitrile using hexamethylsilazane (HMDS) results in the formation of polytrimethylsilyl kanamycin A. The degree of silylation is not yet known, but it is believed that kanamycin A contains 10 silyl groups. Polysilylated kanamycin A contains 10 silyl groups. Polysilylated kanamycin A is well soluble in most organic solvents. Acylation of SAE in anhydrous acetone at 23 ° C and using a 1: 1 molar ratio of SAE to kanamycin produces a mixture containing Cbz-amikacin derivatives and BB-K29, usually in a ratio of 2 to 3: 1. In addition, the composition comprises about 5 to 8% BB-K6, 15 to 20% unreacted kanamycin A, and about 5 to 10% polyacylated material. As in the previous cases, acylation of polytrimethylsilyl 6'-N-benzoxycarbonylcanamycin A could not be detected in any BB-K11 sample. The acylation mixture in acetone was subjected to reduction and conventional work-up followed by CG-50 (N'H4 ⁺ ) column chromatography using 0.5 N ammonium hydroxide to give the isolated crystalline amikacin in 3 to 39% yield.

он O №

kanamycin A (silyl) 10

CisHneOiiNlSiio (1206,35)

В.

он

ChzNH (СN ^ -СН-СООН + HO-N

ВНВА (253.4)

Н — HOS (206.3) (115.9)

EtOAc

DCU + CbzNH (CH3) (2) SAE (350,33) (224,3)

II

C.

Cbz Ammonia (720) © ® ^acetone <

Noc: 2 ° C

Cbz BB-K29

Cbz BB-K6 +

H2 kanamycin A

5% Pd / C polyacylated 1,3-dlBHBA-kanarnycin A aminequinate + BB-K29 + BB-K6 + 1,3-diAHBA-kanamycin A j kanamycin A (585,62) (722,76)

CG-50 (NHf amikacin

III. Materials

Weight (g) Volume (ml) Mol

kanamycin A in free form 50 0,103 HMDS (specific gravity) 0.774) 86.68 112 0.537 acetonitrile 600 UAE 35,03 0.10 acetone 850 CG-50 (NH1 +) 3000 methanol as required IPA as required

IV. Security precautions

Kanamycin A in free form - common safety precautions kanamycin A (silyl) io - no direct information available, common safety precautions other materials - data are given in Example 5.

In the embodiment.

A. A process for producing kanamycin A (silyl) 10

1. 50 g of free kanamycin A (KF · 2.5 to

3.5%) was suspended in 500 ml acetonitrile (KF < 0.01%). The mixture was heated to 74 ° C under reflux while an anhydrous nitrogen stream was passed through the slurry.

2. 112 ml of hexamethyldisilazane (HMDS) are slowly added over 30 minutes. Within 4 to 5 hours, an effective solution was formed with ammonia gas evolving.

3. Heat the mixture at reflux for 22-25 hours under a nitrogen atmosphere.

4. Concentrate the clear pale yellow solution in vacuo at 40 ° C to a syrupy residue.

An additional 100 ml of acetonitrile was then added, the solvent was evaporated and the residue was dried under high vacuum for 3-6 hours. In a yield of 90 to 95% of the theoretical amount, calculated on kanamycin A (silyl) 10, 109 to 115 g of an off-white amorphous powder are obtained.

B. Method for the preparation of L- (-) -. Alpha.-benzyloxycarbonylamino-.alpha.-hydroxybutyric acid N-hydroxysuccinimide ester (SAE)

1. Dissolve 100 g of L- (-) - α -benzyloxycarbonylamino-α-hydroxybutyric acid (BHBA) and 45,38 g of N-hydroxysuccinimide (N-HOS) in 1300 ml of ethyl acetate (KF <0,05 percent) to stirring at 23 ° C.

Second 81.29 g of dicyclohexylcarbodiimide (DCC) were dissolved in 400 ml ethyl acetate (KF <<0.05 ·%) at 23 ^Q C. With vigorous stirring, this solution · added over 30 minutes to a solution of the first step. The temperature rises to 40-42 ° C while precipitating dicyclohexylurea (DCU).

The suspension is stirred for 3-4 hours during which time the temperature has stabilized at 23 ° C.

3. The dicyclohexylurea is separated by filtration, the filter cake is washed with 250 ml of ethyl acetate (KF <0.5%). The filter cake is discarded and the filtrate and washings are processed to the next stage.

4. Concentrate the filtrate and wash to 500 ml in vacuo at 30-35 ° C. A certain amount of solid crystallized out.

5. Transfer the concentrate to a suitable container and add 100 mL of heptane with vigorous stirring. If necessary, small SAE crystals are also added. Crystallization occurs almost immediately. The suspension was stirred for another 30 minutes at 23 ° C.

6. 400 ml of heptane are added over 30 minutes and the suspension is stirred for an additional 4-5 hours at 23 ° C.

7. Collect the solid by filtration and wash the filter cake with 200 mL of heptane / ethyl acetate 3: 1 and then 100 mL of heptane.

8. Dry the mixture under vacuum at 30-35 ^° C for 18-20 hours. 110.1 - 131.4 g of solid are obtained in a yield of 80 to 95%.

Melting point: 119-120 ° C, softening at 114 ° C.

Thin layer chromatography using a 4: 12: 1 mixture of acetone, benzene and acetic acid determined using a 1% aqueous potassium permanganate solution.

Rf is 0.7 pro SAE and 0.2 for BHBA on a 2 x 10 cm silica gel plate (Analtech Inc).

C. Acylation of kanamycline A (silyl) 10

1. Dissolve Kanamycin A (silyl) 10, isolated in Part A, Step 4, in 500 ml of anhydrous acetone at 23 ° C.

2. With vigorous stirring, add 35.03 g of SAE, prepared according to Part B, as a /θ / ο solution in anhydrous acetone over a period of 5 to 10 minutes. The temperature rises to about -5 ° C. The solution was allowed to equilibrate at 23 ° C and further stirred for 18-20. hours.

3. Dilute the light-orange clear solution with 400 ml of water and reduce the pH to 7,0 to 7,5 to 2,2 to 7

2.5 by adding. 3 N hydrochloric acid. The clear solution was stirred at 23 ° C for 15 to 30 minutes.

4. Acetone is removed under vacuum at a bath temperature of 30-35 ° C, sometimes a small amount of material precipitates at this stage, which does not cause further processing. The concentrate is transferred to a suitable dehydrogenation vessel and 10 g of 5% palladium-on-carbon catalyst is added, and the mixture is subjected to hydrogenation at 0.35 MPa for 2-3 hours.

5. Filter the mixture through a Dicalit pad, rinse the hydrogenation vessel and the filter cake with 2 x 50 ml water.

6. Concentrate the filtrate and washings under vacuum at -40 ° ^C to 45 ^° C to approximately one-third of the original volume (150-165 mL).

7. In this step, the mixture has a pH of 6.0 to 7.0. The mixture was applied to a 6 x x 110-cm column containing CG-50 (NH4 ⁺ ).

8. The column is washed with '-1000 ml' of deionized water and then washed with 0.5 N ammonium hydroxide using an automatic polarimeter to monitor elution. The individual components of the mixture are eluted in the following order:

the remaining kanamycin A, BB-K6, BB-K29 and amikacin. BB-K11 was not detectable in any experiment.

9. The fractions containing amikacin are combined and concentrated to contain 25 to 30% solids. The mixture was diluted with an equal volume of methanol and small crystals of amikacin were added to induce crystallization.

10. 2 volumes of IPA are slowly added over 2 hours with vigorous stirring, and crystallization is then carried out for 6 to 8 hours at 23 ° C.

11. The solid is collected by filtration, washed with 35 ml of a 1: 1: 2 mixture of water, methanol and isopropanol and then -35 ml of IPA.

12. The crystals are dried under vacuum at 40 ° C for 16 to 24 hours. This gives 19.91 to 22.84 g of the title compound in a yield of 34 to 39%; MS - infrared, RMR a- ⁿ C NMR spectra and specific -otáčivost are -souhlasu with the structure of the title compound.

Thin layer chromatography

A 1: 4: 2: 1 mixture of chloroform, methanol, ammonium hydroxide and water and 5 x 20 cm silica gel plates (Quantum Industries) was used. One zone of amikacin of Rf 0.4 was detected when detected with ninhydrin.

Example 7

Process for the preparation of amikacin by acylation of poly (trimethylsilyl) -6‘-N-Cbz-kanamycin A in tetrahydrofuran with mixed pivalic anhydride and BHBA

A. Preparation of mixed anhydride

5.06-6 g (20.0 mmol) of BHBA, 4.068 g (20.0 mmol) of triethylamine are dissolved in 200 ml of dehydrated tetrahydrofuran using a molecular sieve. The solution was refluxed for 2 hours 15 minutes, then cooled to -10 ° C. Pivaloyl chloride (2.412 g, 20.0 mmol) was added over 2 to 3 minutes with stirring, and the mixture was stirred at -10 ° C for a further 2 hours. The temperature of the reaction mixture was then allowed to rise to 23 ° C.

B. Acylation of (trimethylsilyl) -6‘-N-Cbz-kanamycin A poles

5.454 g (4.97 mmol) of poly (trimethylsilyl) -6'-N-Cbz-kanamycin A, prepared as described in Example 1, calculated as 6'-Cbz-kanamycin A (silyl) s, are dissolved in 50 ml anhydrous tetrahydrofuran, dried by molecular sieve, at a temperature of 23 ° C. Then half (10.0 mmol) of the mixed anhydride solution prepared is added over 20 minutes. . · According to the above. of said step · A · with vigorous stirring, and then the mixture is further stirred for 7 days.

The reaction mixture was then treated with 100 ml of water and the pH of the original. value was 5.4, adjusted to 2.0 by addition of 3 M · sulfuric acid. The mixture was stirred for an additional hour and then extracted with ethyl acetate. ·. The polyacylated material is highly crystallized so that the reaction mixture must be filtered. After drying over phosphorus pentoxide, 0.702 g of solid is obtained. The reaction mixture was further extracted with a total of 4 x 75 mL of ethyl acetate, then excess ethyl acetate was removed from the aqueous layer by distillation. A sample of the aqueous solution was subjected to high pressure liquid chromatography analysis. The resulting curve shows that the yield of di-Cbz amikacin is 26.4%.

The aqueous layer is then subjected to hydrogenation in a Parr apparatus at a hydrogen pressure of 50 psi using 0.5 g of 10% palladium on charcoal catalyst. The resulting material was collected by filtration and the filtrate and washings were assayed using E. coli to demonstrate that the yield of amikacin was 31.2%. The ratio of amikacin to BB-K29 is approximately 9 to 10: 1, and traces of polyacylated material and unreacted ¹ · kanamycin A were also detected.

Example 8

Effect of water on the preparation of amikacin by acylation of poly (trimethylsilyl) -canamycin A in acetone solution at 23 ° C

A. Anhydrous solvent

2.40 g (2.0 mmol) of poly (trimethylsilyl) kanamycin A, prepared by the method of Example 3, recalculated to kanamycin

A, (silyl) 10 was dissolved in 20 ml of acetone, which had been previously dewatered with a molecular sieve. The solution is stirred at 23 ° C and a solution of 0.701 g (2.0 mmol) of SAE in 10 ml of anhydrous acetone, also dried by molecular sieve, is added. The addition is carried out within 10 seconds. The mixture was then stirred at 23 ° C for 22 hours. 50 ml of water are added and the initial pH of 7.5 is adjusted to 2.5. The acetone is removed by distillation under vacuum at 40 ° C and the aqueous solution is then reduced under a hydrogen pressure of 0.35 MPa at 23 ° C for 2 hours as catalyst. 1 g of 10% palladium on activated carbon is used. A microbiological experiment showed 31.24% of amikacin.

B. Add water to solvent

Step A was repeated except that 1.0 ml (56 mmol) of water was added to the poly (trimethylsilyl) kanamycin A solution and the mixture was stirred for 15 minutes before the SAE acylation. · Microbiological. tests showed that the yield of amykacin in this case was 33.80%.

Example 1 and d · 9. .

Process for the production of amikacin. acylation of · poly (trimethylsilyl) -6‘-N-Cbz-kanamycin A in acetone mixed anhydride · BHBA · and isobutylcarboxylic acid

A. Preparation of mixed anhydride

1.267 g (5.0 mmol) of BHBA and 1.333 g (10.0 mmol) of N-trimethylsilylacetamide (MSA) in 20 ml of molecular sieve acetone are stirred at 23 ° C with the addition of 0.70 g. ml (5.0 mmol) of trimethylamine (TEA). The mixture was heated to reflux under a nitrogen atmosphere for 2.5 hours. The mixture was cooled to -20 ° C and isobutyl chloroformate (0.751 g, 0.713 mL, 5.50 mmol) was added. Triethylamine hydrochloride precipitated immediately. The mixture was then stirred at -20 ° C for 1 hour.

В. Acylation

6.215 g (4.9 mmol) of poly (trimethylsilyl) -6'-N / Cbz-kanamycin A, prepared as described in Example 1, recalculated to a compound with 9 forces of 1-group, are dissolved in 20 ml of acetone, previously dehydrated with molecular sieve with stirring at 23 ° C. The solution was cooled to -20 ° C and then the cooled mixed anhydride solution from Step A was slowly added over 30 minutes. The reaction mixture was then stirred at -20 ° C for 1.5 hours and then at 23 ° C for 17 hours. C. The reaction mixture was poured into 150 ml of water at 23 ° C under stirring, and the initial pH of 7.75 was adjusted to 2.5 with 3 N _t -alkyl hydrochloric, and the mixture stirred for a further 15 minutes. The acetone was then removed by distillation under vacuum at 40 ° C. A sample of the resulting solid solution was subjected to high pressure liquid chromatography. The curve shows that the yield of di-Cbz-amikacin was 34.33%.

A major portion of the aqueous solution obtained above is subjected to a reduction under a hydrogen pressure of 50 psi at 23 DEG C. for 3 hours and 15 minutes, using 2.0 g of palladium on charcoal as the catalyst. The catalyst was separated by filtration, the filtrate and washings were combined and the activity determined microbiologically against E. coli, indicating that the yield of amikacin was 35.0 percent.

Example 10

Process for the preparation of amikacin by acylation of (trimethylsilyl) -6'-N-Cbz-kanamycin A poles in 3-pentanone g (23.65 mmol) with (trimethylsilyl) -6'-N-Cbz-kanamycin A poles prepared according to Example 1 recalculated to 6'-N-Cbz-kanamycin A (silyl) 9 in solution in 100 ml of 3-pentanone, previously dehydrated by means of a molecular sieve, is stirred at 23 ° C with 26.02 added over 40 minutes mmol, ie ΙΟθ / o excess

NAE. The mixture was stirred at 23 ° C for 113 hours and added to 250 ml of water with vigorous stirring. The original pH of 7.3 was adjusted to 2.5 by the addition of 3 N hydrochloric acid as the mixture was stirred for an additional 30 minutes and the 3-pentanone was removed by distillation under vacuum at 40 ^° C. The aqueous solution was extracted with 4 x 100 ml. ethyl acetate. A sample of the aqueous solution is then subjected to high pressure liquid chromatography. The resulting curve showed that the yield of di-Cbz-amikacin was 46.12%.

The bulk of the aqueous reaction mixture was subjected to a reduction under a hydrogen pressure of 50 psi at 23 ° C for 2.5 hours using 3 g of 10% palladium on charcoal as the catalyst. The filtrate and washings are combined and a sample of this material is subjected to microbiological tests to demonstrate that the yield of amikacin is 40.24%. The bulk of the reduced aqueous reaction mixture was then concentrated in vacuo at 40 ° C to a volume of approximately 100 mL and the material was applied to a 130 x 10 cm column containing approximately 10 liters of CG-50 (NH 4 ⁺ ). After the aqueous solution has been applied to the top of the column, the column is washed with 5 liters of water and the active substance is then eluted first with 0.5 N ammonium hydroxide, then the polyacylated products are washed off with 3 N ammonium hydroxide. Polarimetry of the individual fractions showed a yield of 42.7% amikacin, 12.0% unreacted kanamycin A, 12.4% polyacylated material and 23.2% BB-K29.

Example 11

Process for the preparation of amikacin by acylation of poly (trimethylsilyl) -6‘-N-Cbz-kanamycin A in anhydrous cyclohexanone at different times

A. 2.537 g (2 mmol) of poly (trimethylsilyl) -6'-N-Cbz-kanamycin A prepared as described in Example 1, calculated on 6'-N-Cbz-kanamycin A (silyl) 9 in 300 ml of anhydrous cyclohexanone, acylated at 23 ° C for 20 hours with a solution of 10.8 ml (0.1944 mmol / ml, 2.10 mmol) of NAE in anhydrous cyclohexanone. The reaction mixture was then added to 150 ml of water with stirring and the initial pH of 5.6 was adjusted to 2.5 by addition of 3 N hydrochloric acid. The cyclohexanone was removed by distillation under vacuum at 40 ° C and a sample of the remaining aqueous phase was taken for high pressure liquid chromatography testing.

The major portion of the aqueous phase was reduced at a hydrogen pressure of 50 psi for 3 hours at 23 ° C using 1 g of 10% palladium on charcoal as catalyst. The catalyst is then removed by filtration, the filtrate and washings are combined and subjected to a microbiological test for amikacin.

B. The above reaction A was repeated except that the acylation was carried out for 115 hours instead of 20 hours.

Yields

Liquid high pressure chromatography (di-Cbz amikacin) test

Microbiological test for amikacin turbidimetrically on reaction plate A 49,18% reaction B 56,17%

Example 12

Process for the preparation of amikacin by acylation of (trimethylsilyl) -6'-N-Cbz-kanamycin A in anhydrous tetrahydrofuran at different times

A. Example 11 is repeated except that

Yield Liquid HPLC test (di-Cbz amikacin) Reaction A 2-9.27% Reaction B 233.99%

Example 13

A process for the production of amikacin by acylation of poly (trimethylsilyl) -6 N-N-Cbz-kanamy tin A in anhydrous dioxane at different times

42.87% 39.16 O / o

55.39% 38.45% that anhydrous tetrahydrofuran is used as the solvent instead of anhydrous cyclohexanone.

B. Example 11B is repeated except that anhydrous tetrahydrofuran is used as the solvent instead of anhydrous cyclohexanone.

€

Microbiological test for amikacin turbidimetrically on plates

28,34% 28,18%

21,52% 28,63%

A. Example 11A was repeated except that the acylation was carried out for 44 hours using anhydrous dioxane as the solvent.

B. Example 11B was repeated except that the acylation was carried out for 18.5 hours using anhydrous dioxane as solvent.

Yield Liquid high pressure chromatography (di-Cbz amikacin) reaction A reaction B

39,18%

42,82%

Example 14

A process for the production of amikacin by acylation of (trimethylsilyl) -6'-N-Cbz-kanamycin A in anhydrous diethyl ketone at 75 ° C

To 2.537 g (2.0 mmol) of poly (trimethylsilyl) -6'-N-Cbz-kanamycin A, prepared as described in Example 1, converted to 6'-N-Cbz-kanamycin A (silyl) 9 in 32 ml · of diethyl ketone, previously dehydrated by molecular sieve, is added, with stirring at 75 ° C, 10.8 ml (0.1944 mmol / ml, 2.10 mmol) of the NAE solution in diethyl ketone over 15 minutes. The mixture was further stirred for 3 hours at 75 ^DEG C., then poured into 150 ml of water ·. The pH was adjusted to pH

2.8 N hydrochloric acid and diethyl ketone were removed by distillation under vacuum at 40 ° C. High pressure chromatography of the aqueous phase sample indicated a yield of di-Cbz of amikacin of 39.18%.

The major portion of the aqueous phase was reduced at a hydrogen pressure of 50 psi for 3 hours 15 min at 23 ° C using 1.0 g of palladium on charcoal as catalyst. The catalyst was removed by filtration, the filtrate

Microbiological test for amikacin turbidimetrically on plates

43.27 o / o 33.36%

22.55 O / o 33.37 ·% and the wash material is then subjected to microbiological tests for amikacin. In the turbidimetric test it was possible to show a result of 27.84% in the plate test a yield of 28.6%.

Example 1 - ad 15

A process for the preparation of amikacin by acylation of (trimethylsilyl) kanamycin A poles.

by treatment with · NAE at 0 to 5 ° C after back hydrolysis with water

A. Silylation of kanamycin A using HDMS together with TMCS as a catalyst g (20.14 mmol) of kanamycin A having a purity of 97.6% in 100 ml of acetonitrile previously dewatered by molecular sieving was heated to reflux in nitrogen. atmosphere. A mixture of 22.76 g (141 mmol, i.e. 7 moles per 1 mol mol of kanamycin A) HMDS and 1 ml (0.856 g, 7.88 mmol) of TMCS is added to the reaction mixture over 10 minutes while boiling. . The mixture is then further refluxed for 4 hours and 45 minutes, then cooled, concentrated in vacuo to a viscous yellow syrupy liquid and then dried under high vacuum for 2 hours. The product was obtained in a yield of 97.9 percent in an amount of 23.8 grams, calculated as kanamycin A (strength 10%).

B. Acylation

23.8 g (20.14 mmol) of poly (trimethylsilyl) -canamycin A prepared in step A are dissolved in 250 ml of acetone, previously dehydrated by molecular sieving at 23 ° C, after which the solution is cooled to 0 to 5 ° C, vigorously. 3.63 ml (201.4 mmol) of water, i.e. 10 moles of water per 1 mol of polysilylated kanamycin A, are added with stirring and the mixture is allowed to stand at a slight vacuum for 30 minutes, then 19.133 mmol are added in less than 1 minute, i.e. 0.95 mole per mole of polysilylated kanamycin A NAE in 108.3 mL of acetone, stirred at 0-5 ° C for one hour, then diluted with water, adjusted to pH 2.5 and the acetone was removed in The aqueous solution was then reduced under a hydrogen pressure of 0.35 MPa at 23 ° C and 2.5 hours using 2.0 g of 10% palladium on charcoal as a catalyst.The filtered reaction mixture was filtered through a pad of Dicalit, concentrated. in vacuo to a volume of approximately 100 mL at 40 ^° C and then applied to a 5 x 100 cm column containing 5 liters of CG-50 (NH4 ⁺ ). The column was washed with water and then eluted sequentially with 0.6 N, 1.0 N, and 3 N ammonium hydroxide. 60.25% of amikacin, 4.37% of BB-K6, 4.35% of BB-K29, 26.47% of kanamycin A and 2.18% of polyacylated products are obtained.

Example 16

Process for the production of amikacin by acylation of (trimethylsilyl) -6‘-N-Cbz-kanamycin A with SAE at 0-5 ° C after reverse methanolysis

A. Silylation of 6‘-N-Cbz-kanamycin A

20.0 g (32.4 mmol) of 6‘-N-Cbz-kanamycin A in 200 ml of acetonitrile, previously dehydrated by molecular sieving, are heated to reflux under a nitrogen atmosphere. Then 47.3 ml (226.8 mmol) of HDMS, i.e. 7 moles of this substance per 1 mol of 6'-N-Cbz-kanamycin A are added over 10 minutes and the mixture is then heated to under reflux for 20 hours. The reaction mixture was cooled, concentrated in vacuo and dried under high vacuum for 2 hours to give 39.1 g of a white amorphous solid. The yield is 95.4%, calculated as 6‘-N-Cbz-kanamycin A (silyl) 9.

B. Acylation

39.1 g (32.5 mmol) of poly (trimethylsilyl) -6'-N-Cbz-kanamycin A, prepared according to Step A, are dissolved in 400 ml of anhydrous acetone with stirring at 23 ° C. Then, 6.6 mL (162 mmol) of methanol, i.e. 5 moles of methanol per mole of polysilylated 6'-N-Cbz-kanamycin A, was added and the mixture was stirred at 23 ° C for one hour in a strong stream of nitrogen. The mixture was cooled to 0-5 ° C and a solution of 11.35 g, 32.4 mmol, SAE in 120 mL of pre-cooled anhydrous acetone was added. The reaction mixture was stirred for an additional 3 hours at 0-5 ° C and then stored in a cold room at 4 ° C for 1 week. 300 ml of water are then added, the pH is adjusted to 2.0, the mixture is stirred for an additional hour and then the acetone is removed by distillation in vacuo. The resulting aqueous solution was reduced under a hydrogen pressure of 50 psi for 17 hours at 23 ° C using 3.0 g of 10% palladium on charcoal as catalyst. The resulting reaction mixture was filtered through a Dicalit pad, then concentrated in vacuo to a volume of 75-100 ml and applied to a CG-50 (NH4 ⁺ ) column, then eluted with water and 0.6 N ammonium hydroxide. · Tíipto is obtained 52.52% amikacin, 14.5% BB-K29, 19.6% of kanamycin A and 1.71 v _/ polyacylovaných materials.

Example 17

A process for the production of amikacin by acylation of the (trimethylsilyl) kanamycin A poles with SAE at 0-5 ^° C after back hydrolysis with water

A. Kanamycin A silylation by TMCS in acetonitrile using tetramethylguanidine as acid binding agent

4.88 g (10.07 mmol) of kanamycin A are suspended in 100 ml of acetonitrile previously dewatered by molecular sieving with stirring at 23 ° C. 16.234 g (140.98 mmol) of tetramethylguanidine (TMG), i.e. 14 moles of this substance per mole of kanamycin A, are added to the suspension by stirring. The mixture is heated to reflux and 15.32 are added over 15 minutes. g, (140.98 mmol) of TMCS, i.e. 14 moles of this substance per mole of kanamycin A. A white TMG hydrochloride precipitate formed when half of the TMCS was added. The mixture was cooled to room temperature, concentrated to a sticky residue and dried under high vacuum for 2 hours. The solid obtained is triturated with 100 ml of anhydrous tetrahydrofuran and the insoluble TMG hydrochloride is collected by filtration and washed with 5 x 20 ml of tetrahydrofuran. The filtrate and washings were combined and concentrated in vacuo at 40 ° C to a sticky residue, then dried under high vacuum for 2 hours. This gives 10.64 g of the cream world le sticky residue, yield 87.6 o / _o, based on kanamycin A (silyl) io.

B. Acylation

10.64 g (10.07 mmol) of the (trimethylsilyl) kanamycin A poles, introduced in step A, are dissolved in 110 ml of acetone, previously dewatered by molecular sieves, with stirring at 23 ° C and the solution is cooled to 0 to 5 ° С. Then 1.81 ml (100.7 mmol) of water, i.e. 10 moles of water per 1 mol of polysilylated kanamycin A, are added, and the solution is stirred under gentle vacuum for 30 minutes. Then add in less than 1 minute

3.70 g (10.57 mmol), i.e. a 5% excess SAE in 40 ml of pre-cooled anhydrous acetone, and the mixture was further stirred for an hour. The mixture is then further processed as in Example 16B to give about 50% amikacin, 10% BB-K29, 5-8% BB-K6, 20% kanamycin A, and 5-8% polyacylated products. .

Example 18

Process for the preparation of kanamycin A (trimethylsilyl) poles in pyridine using HMDS

10.0 g (20.64 mmol) of kanamycin A are suspended in 100 ml of dehydrated pyridine by molecular sieving at 23 ° C. Nitrogen is then passed through the mixture and the suspension is heated to reflux. Then, 17.33 g (107.32 mmol) of HMDS, ie.

5.2 moles of this substance per mole of kanamycin A and the mixture was further heated at reflux for 19 hours. The mixture was cooled to room temperature, concentrated in vacuo to a pale golden yellow syrup liquid, which was then dried under high vacuum to a white amorphous solid. 22.1 g of product, calculated on kanamycin A (silyl), are obtained in a yield of 92.6%.

Example 19

Process for the preparation of kanamycin A poly (trimethylsilyl) using triethylchlorosilane and triethylamine as acid binding agent

5.0 g (10.07 mmol) of kanamycin are suspended in 100 ml of acetonitrile previously dewatered with molecular sieves at 23 DEG C. to a purity of 9.6%. Then, 33.8 ml, 24.5 g, (241.7 mmol) of triethylamine (TEA) are added and the suspension is heated to reflux. A solution of 23.7 ml, 21.3 g (111.98 mmol) of trichloroethylsilane in 25 ml of anhydrous acetonitrile is then added over 20 minutes. The mixture was heated at reflux for an additional 7 hours, then cooled to room temperature to precipitate TEA hydrochloride as long fine needles. The mixture was allowed to stand at room temperature for approximately 16 hours, then concentrated in vacuo at 40 ° C to a sticky solid which was dried under high vacuum for 2 hours to a deep orange sticky solid. This material was triturated with 100 mL of anhydrous tetrahydrofuran at 23 ° C and the insoluble TEA hydrochloride was collected by filtration, and dried to give 16.0 g of TEA hydrochloride. The filtrate and the wash liquid were dried under vacuum to a solid which was then dried under high vacuum for 2 hours. In this way it is obtained

19.3 g (triethylsilyl) of kanamycin A in the form of a strong orange viscous syrupy liquid.

Example 20

Process for the preparation of kanamycin A poles (trimethylsilyl) using bis-trimethylsilyl urea

10.0 g (20.58 mmol) of kanamycin A having a purity of 99.7% are suspended in 200 ml of acetonitrile previously dewatered by molecular sieving with stirring at 23 ° C. 29.45 g (144.01 mmol) of bis-trimethylsilylurea (BSU), i.e. 7 moles of this substance per mole of kanamycin A, are added to the suspension, then the mixture is refluxed under a nitrogen atmosphere. Heating is continued for 17 hours, then the reaction mixture is cooled to room temperature. A small portion of the insoluble material was collected by filtration. It is washed with 3 x 10 ml of acetonitrile and dried to give 1.1381 g of insoluble solid. Infrared light shows that it is BSU and a small amount of unreacted kanamycin A. The filtrate and washings are combined and stored at 4 ° C for 16 hours. In this way, an additional solid is collected which is isolated as above, yielding 7.8 grams of this solid, consisting of BSU and urea, as shown by infrared light. The light yellow filtrate and washings were concentrated in vacuo at 40 ° C and dried under high vacuum to give 27.0 g of a white solid which was partly in the form of white fine needle crystals and partly in the form of a sticky solid. This material was treated by adding 150 mL of heptane at 23 ° C, insoluble matter was collected by filtration, washed with 2 x 50 mL of heptane and then dried to give 6.0 g of white needle crystals. It can be shown in the infrared light that it is a mixture of BSU and urea. The filtrate and washings were combined, concentrated in vacuo at 40 ° C and dried under high vacuum for 2 hours to give 20.4 g of white needle crystals. It can be shown in infrared light that the spectrum is typical of polysilylated kanamycin A. It has been shown by count that the product content is on average 7.22 trimethylsilyl groups.

Example 21

Process for the production of amikacin by acylation per (trimethylsilyl) kanamycin A after partial desilylation by treatment

1,3-butanediol

Preparation of per (trimethylsilyl) kanamycin

10.0 g (20.639 mmol) of kanamycin A are suspended in 100 ml of dewatered acetonitrile by molecular sieving with stirring at 23 ° C. The suspension is heated to reflux under a stream of nitrogen, whereupon the 23.322 g (144.5 mmol) of HMDS, which is 7 moles of this substance per mole of kanamycin A, are added over 10 minutes. It is cooled to room temperature, concentrated in vacuo and dried under high vacuum for 2 hours. 24.3 g of a white sticky substance are obtained in a yield of 92.1% based on kanamycin A (silyl.

B. Acylation

24.3 g of per (trimethylsilyl) kanamycin A prepared according to step A are dissolved with stirring in 240 ml of dehydrated water by molecular sieving at 23 ° C. To this solution is added 9.25 ml (103.2 mmol) of 1,3-butanediol, i.e. 5 moles of this substance per mole of per (trimethylsilyl) kanamycin A. The mixture is then stirred at 23 ° C for 2 hours. a stream of nitrogen and then cooled to 0-5 [deg.] C. A solution of 7.23 g (20.64 mmol) of SAE in 70 ml of pre-cooled acetone is added over 1 minute and the mixture is stirred at room temperature for 3 hours. The mixture is allowed to stand for about 16 hours in a cold room at 4 DEG C. Then 200 ml of water are added, the pH is adjusted to 2.5 and the clear yellow solution is stirred at 23 DEG C. for 30 minutes. Acetone was removed by filtration under vacuum and the aqueous solution was reduced at 0.39 MPa of hydrogen at 23 ° C for 2 hours, using 3.0 g of 10% palladium on charcoal as the catalyst. Dicalite and chromatographed as in Example 16B to yield ·

47.50 o / o amikacin, 5.87% BB-K29, 7.32% BB-K6, 24.26% kanamycin A, and 7.41% polyacylated products.

Example 22

Process for making amikacin by acylating poly (trimethylsilyl) kanamycin A prepared in tetrahydrofuran using SAE and sulfamic acid as catalyst

To a mixture of 5.0 g (10.32 mmol) of kanamycin A and 50 ml of tetrahydrofuran (THF) previously dehydrated by molecular sieving, 100 mg of sulfonic acid and 12.32 g (76.33 mmol) of ethanol are added at reflux. The mixture was heated to reflux for 18 hours, resulting in a clear solution after 6 hours, cooled to 23 ° C, added with 0.1 mL of water and held for a further 30 minutes. at 23 DEG C. A solution of 3.61 g (10.3 mmol) of SAE in 36 ml of tetrahydrofuran is then added over 30 minutes, the mixture is stirred for a further 3 hours and then diluted with 100 ml of water and adjusted to pH 10%. The mixture was stirred at 23 ° C for 30 minutes, then the tetrahydrofuran was removed in vacuo, and the resulting aqueous solution was reduced at 50 psi of hydrogen for 2 hours at 23 ° C. ° C, using 10% palladium on activated carbon as the catalyst The filtrate and washings in a total amount of 150 ml are subjected to microbiological tests against E. coli, yielding 1225 m. of amikacin programs in a yield of 31.5%. ml of material.

Example 23

Process for the preparation of amikacin by acylation of poly (trimethylsilyl) kanamycin A with dibenzyloxycarbonyl N-hydroxysuccinimide ester AHBA

A. Preparation of dibenzyloxycarbonyl-L- (-) -. Alpha.-amino-.alpha.-hydroxybutyric acid in the form of N-hydroxysuccinimide ester g (20.65 mmol) of dibenzyloxycarbonyl-L- [-] -. Alpha.-amino-.alpha.-hydroxybutyric acid .alpha. 37 g (20.65 mmol) of N-hydroxysuccinimide are dissolved in 50 ml of anhydrous acetone at 23 DEG C. Then 4.25 g (20.65 mmol) of dicyclohexylcarbodiimide in solution in 20 ml of anhydrous acetone are added and the resulting mixture The mixture is stirred for 2 hours at 23 DEG C. The dicyclohexylurea is separated by filtration, the filter cake is washed with 10 ml of anhydrous acetone and the filtrate and washings are combined.

B. Acylation

Poly (trimethylsilyl) kanamycin A prepared as described in Example 21 from 10.0 g (20.639 mmol) of kanamycin A was dissolved in 100 mL of anhydrous acetone, cooled to 0-5 ° C, 3.7 was added. ml of deionized water and the solution is stirred at 0-5 ° C for 30 minutes under slight vacuum.

To the solution is added a solution of the di-Cbz-protected acylating agent prepared in Step A, and the mixture is stirred for 30 minutes at a temperature of 0 to 5 ° C. The mixture was diluted with water, adjusted to pH 2.2 and the acetone removed in vacuo. The aqueous solution was reduced as in Example 22 and filtered with a Dicalit layer. Chromatography showed 40-45% amikacin, approximately 10% BB-K29, traces of BB-K6, approximately 30% kanamycin A. and a small amount of polyacylated materials.

Example 24

Process for the preparation of kanamycin A poly (trimethylsilyl) using HMDS and imidazole as catalyst g (22.7 mmol) of kanamycin A and 100 ml of imidazole are heated to reflux in 100 ml of acetonitrile, dehydrated by molecular sieving in a stream of nitrogen. . Then 18.48 g (114.5 mmol) of MHDS, which is 5 moles per 1 mol of kanamycin A, are added over 30 minutes and the mixture is further refluxed for 20 hours. A clear solution is formed in about 2.5 hours. The solution was cooled to 23 ° C and the solvent was evaporated in vacuo to give 21.8 g of poly (trimethylsilyl) kanamycin A as a foamy residue. The yield is 93.1%, based on kanamycin A (silyl) n.

Example 25

Process for the preparation of 1-N- (L- (-) -? -Amino-.alpha.-hydroxybutyryl) cyanamycin B (BB-K26) by acylation of poly (trimethylsilyl) cyanamycin B by treatment with

A. Preparation of poly (trimethylsilyl) kanamycin B by treatment. HMDS using TMCS as catalyst g (51.7 mmol) of kanamycin B in 250 ml of dewatered acetonitrile by molecular sieving is heated to reflux. in a stream of nitrogen. Then. 62.3 g (358.81 mmol) of HMDS, i.e. 7.5 moles of this substance per mole of kanamycin B, are added over 30 minutes, after which 1 ml of TMCS catalyst is added. The mixture was heated at reflux for 21 hours, resulting in a clear solution after 1 hour. The solvent was evaporated in vacuo at 60 ° C and the oily residue was kept at 60 ° C under high vacuum for 3 hours. In this way, 53.0 g of poly (trimethylsilyl) kanamycin B are obtained. The yield is 85.2% based on kanamycin B (silyl) 10.

B. Acylation

The poly (trimethylsilyl) -canamycin B, prepared in Step A at 53.0 g, is dissolved in 500 ml of anhydrous acetone at 0-5 ° C, 20.9 ml of methanol are added and the mixture is stirred for 30 minutes · under vacuum at 0-5 ° C. A solution of 18.1 g (51.67 mmol) of SAE in 200 ml of pre-cooled anhydrous acetone is then added over a period of less than 1 minute, and the mixture is stirred for a further 30 minutes at 0-5 ° C. The mixture was then treated as in Example 22 and applied to a 6 x 120 cm column containing CG-50 (NH4 ⁺ ). The column is eluted with ammonium hydroxide at an increasing concentration of 0.6 N to 3 N. This gives 38% BB-K26, 5% corresponding to 6'-N-acylated kanamycin B (BB-K22), 10% corresponding to 3-N-acylated kanamycin B (BB-K46) of 14.63 / ₀ kanamycin B and kanamycin polyacylovaného small amount B.

Example 26

Process for producing poly (trimethylsilyl) kanamycin A using HMDS and kanamycin A in the form of sulfate as a catalyst

19.5 g · (40.246 mmol) of kanamycin A and 0.5 g (0.858 mmol) of kanamycin A-sulphate, total 20.0 g (41.0 mmol) in 200 mL of dehydrated molecular sieve are heated to boiling under reflux. Then, 60.3 ml (287.7 mmol) of HMDS, i.e. Ί moles of this substance per 1 mol of kanamycin A, are added slowly and the mixture is stirred. at reflux temperature for 28 hours. The mixture was evaporated to dryness on a rotary evaporator and dried. 47.5 g of poly (trimethylsilyl) kanamycin A are obtained in the form of a pale yellow oil. The yield is 95.82% based on kanamycin A (silyl.

Example '27

Process for the preparation of amikacin by acylation. (N-hydroxysuccinimide ester AHBA, with N-trifluoroacetyl group as protecting group)

A. Preparation of N-trifluoroacetyl AHBA and conversion of this compound to N-hydroxysuccinimide ester

To a suspension of 5.0 g (42 mmol) of AHBA in 100 ml of tetrahydrofuran is added 40 g (191 mmol) of trifluoroacetic anhydride with stirring for 10 minutes. The solution was stirred at 23 ° C for 18 hours and then evaporated to dryness under vacuum at 50 ° C. The residue is dissolved in 100 ml of a 1: 1 mixture of water and methanol and stirred for 1 hour. The mixture is evaporated to dryness in vacuo and redissolved in 50 ml of water. The aqueous solution was extracted with 3 x 50 mL of MIBK and, after drying over sodium sulfate, the extract was concentrated to an oil. The traces of solvent were removed by adding 4 ml of water followed by distillation. On standing, the liquid was changed to 2.5 g of a waxy crystalline solid. Yield 28%.

2.4 g (11.3 mmol) of N-trifluoroacetyl AHBA are dissolved in 50 ml of anhydrous acetone and 1.30 g (11.31 mmol) of N-hydroxysuccinimide are added. A solution of 2.33 g of dicyclohexylcarbodiimide in 20 ml of anhydrous acetone is then slowly added. The reaction mixture was stirred at 23 ° C for 2 hours and the precipitated dicyclohexylurea was removed by filtration and washed with a small amount of acetone. Combine the filtrate and wash. This is a solution of N-hydroxysuccinimide ester N-trifluoroacetyl -AHBA which is used without isolation in the next step.

B. Acylation

To 11.31 mmol of a solution of poly (trimethylsilyl) kanamycin A prepared as described in Example 26 in 54 ml of acetone was added 2.0 ml (113.4 mmol) of water and the mixture was stirred under vacuum at 0-5 ° C. C for 30 minutes. 11.31 mmol of N-trifluoroacetyl AHBA, N-trifluoroacetyl AHBA, prepared in Step A, are then added to the mixture and the mixture is maintained at 5 ° C for 1 hour. The pH is then adjusted. to 2.0 by adding 20%. The mixture was stirred for a further 30 minutes and then adjusted to pH 6.0 by addition of ammonium hydroxide. The mixture was then evaporated to dryness in a rotary evaporator to give 14.4 g of a sticky off-white solid. This material is dissolved in 100 ml of water, the pH is adjusted from 5.5 to 11.0 by adding 10 N ammonium hydroxide and the solution is heated in an oil bath at 70 ° C for 1 hour. The solution was then adjusted from pH 9.5 to pH 7.0 by addition of hydrochloric acid, filtered to remove a small insoluble matter, and the filter was washed with water. The filtrate and washings in a total volume of 188 ml are applied to a 8 × 90 cm column containing CG-50 (NH 4 +), washed with 2 liters of water and eluted with ammonium hydroxide at an increasing concentration of 0.6 N to 1.0 N. In this way, 28.9% amikacin, 5.0% BB-K6, 5.7% BB-K29, 43.8% kanamycin A, 3.25% polyacylated material are obtained. 3% of unknown substance present in the first fraction.

Example 28

A process for the preparation of amikacin by acylation of (poly (trimethylsilyl) kanamycin A with N-hydroxysuccinimide ester of tert-butyloxycarbonyl AHBA

A. Preparation of tert-butyloxycarbonyl AHBA and conversion of this substance to N-hydroxysuccinimidester

5.0 g (42 mmol) of AHBA in 100 ml of water and 20 ml of acetone were adjusted to pH 10 by addition of 10 N sodium hydroxide. . 11.4 g (53 mmol) of di-tert-butyl dicarbonate are added over 3-4 minutes, the solution is stirred for 35 minutes and the pH is maintained at 10 by addition of 10N sodium hydroxide. The acetone is removed in vacuo and the aqueous phase is washed with 40 ml of ethyl acetate. The aqueous solution was adjusted to pH 2.0 by addition of 3 N hydrochloric acid and then extracted with 3.times.30 ml of MIBK. The extracts are combined, dried over sodium sulfate and concentrated to a clear oily liquid. Yield 8.2 g (89%) of the liquid.

4.25 g (19.4 mmol) of t-butyloxycarbonyl AHBA are dissolved in 50 ml of acetone and 2.23 g (19.4 mmol) of N-hydroxysuccinimide are added. A solution of dicyclohexylcarbodiimide (4.00 g, 19.4 mmol) in acetone (20 ml) was added slowly and the mixture stirred at 23 ° C for 2 hours. The precipitated dicyclohexylurea is filtered off and washed with a small amount of acetone. The filtrate and the washing liquid forming the t-butyloxycarbonyl AHBA N-hydroxysuccinimide ester solution were used without isolation in the next step.

B. Acylation

To a solution of 41.28 mmol of poly (trimethylsilyl) kanamycin A prepared as described in Example 26 in 94 ml of acetone is added

3.5 ml (194 mmol) of water and the mixture was stirred under vacuum at 0-5 ° C for 30 minutes. 19.4 mmol of tert-butyloxycarbonyl AHBA prepared in Step A (19.4 mmol) are then added and the mixture is left to stand for one hour at 5 ° C. Then 200 ml of water are added and the initial pH of 7.0 is added. reduced to 2.0 by adding 20% sulfuric acid. ·. After stirring for 30 minutes, the pH was readjusted to 6.0 with ammonium hydroxide and the mixture was evaporated to dryness in vacuo to give 36.3 g of a golden yellow oil. This liquid was dissolved in 200 mL of trifluoroacetic acid, allowed to stand for 15 minutes and then evaporated to dryness in a rotary evaporator. The oily material was washed with water and the water was evaporated. Concentrated ammonium hydroxide is added to pH 6.0 and also evaporated rapidly. The resulting solid was dissolved in water, filtered and the filter washed with water. The filtrate and washings in a total of 259. ml are applied to a 8 x 92 cm column packed with CG-50 (NH4 ⁺ ), washed with 4 liters of water, then eluted with ammonium hydroxide at an increasing concentration of 0 This gives 40.32% amikacin, 4.58% BB-K6,

8.32% BB-K29, 30.50% kanamycin A and

7.43% polyacylated materials.

Example 29

The procedure of Example 1 was repeated except that an equimolar amount of 6'-N-benzyloxycarbonylkanamycin B was used in place of 6'-N-benzyloxycarbonylanamycin A to give 1 N- [L- (- Jy-amino-α-hydroxybutyryl) ] kanamycin B.

Example 30

The method of Example 1 was repeated except that instead of. L- (-) - γ-Benzyloxycarbonylamino-α-hydroxybutyric acid N-hydroxy-5-norbornene-2,3-dicarboximide ester

L - (-) - β-Benzyloxycarbonylamino-α-hydroxypropionic acid N-hydroxy-5-norbenzene-2,3-dicarboximide ester and

L-(-) -? - benzyloxycarbonylamino-α-hydroxyvaleric acid N-hydroxy-5-norbornene-2,3-dicarboximide ester to give

1-N- [L- (-) - N-amino-α-hydroxypropionyl] kanamycin A a

1-N- [L- (-) -. Alpha.-amino-.alpha.-hydroxy-valeryl] kanamycin A.

Example 31

Repeats. The process according to Example 25, with the difference that the N-hydroxy ester of L- (-) -? -benzyloxycarbonylamino-α-hydroxybutyric acid used. .substitutes

L-(-) -? - benzyloxycarbonylamino-α-hydroxypropionic acid N-hydroxysuccinimide ester; and

L-(-) -? - benzyloxycarbonylamino-α-hydroxyvaleric acid N-hydroxysuccinimide ester, thereby. yielding 1-N- [L- (-) - N - (R) - O - (-) - β-β-β-β-β-propionyl} -canamycin. B · a

1-N- [L- (-) -. Alpha.-amino-.alpha.-hydroxy-valeryl] kanamycin B.

Example 1-a d 3 2 .................... ....... ........

Production of BB-K8 by acylation of poly (trimethylsilyl) -3,6‘-di-N-carbobenzyloxycaranamycin A in anhydrous diethyl ketone

A. 3,6‘-di-N-benzyloxycarbonylkanamycin A

Suspension 7.26 g. (15 mmol) of kanamycin A in free form and 18.6 g (75 mmol) of nickel tetraacetate tetrahydrate in 300 ml of dimethylsulfoxide were heated at 100 ° C for 30 minutes with stirring to give a clear, green solution. After cooling, 11.8 g (37.6 mmol) of N-benzyloxycarbonyl-15-n-orborneri-2,3-dicarboximide in 50 ml of dimethylsulfoyide are added to this solution. The mixture was stirred overnight at room temperature, 100 ml of concentrated aqueous ammonia and 1 liter of water were added, the mixture was further stirred at room temperature for an hour, then applied to the top of a column containing 300 ml of Diaion HP-10 resin. . This column is then eluted successively with 7 N ammonium hydroxide, then with a 1: 1 mixture of methanol and water and finally with a 10: 1 mixture of methanol and water, collecting fractions of - 20 ml and monitoring these fractions. thin layer chromatography on silica gel (Merck Silica Gel 60 F-254) using a 1: 2: 1 mixture of chloroform, ethanol and 28% ammonium hydroxide as solvent. A portion of the resulting product which crystallizes as fine needles from the highly concentrated fraction is collected by filtration to give an analogous sample. The filtrate and other fractions containing the resulting product (Rf 0.42) were combined and the solution was evaporated in vacuo. The residue was triturated with diethyl ether to give a total yield of 9.7 g (86%), m.p. &gt; 300 ° C.

IR (KBr) Vmax 1690 cm ^1st

NMR Spectrum · (DMSO-d 6 + DC1, pD = 3):

4.76 to 5.26 (6H, m, Hi‘, Hi “and CO — OCHa — C & H5 x 2),

7.26 (10H, s, CO-OCHa-C 1 H 5 x 2). '

For Ό34Η4 »Ν4θΐ1. HaO calculated:

% H, 6.54;% N, 7.27;

found ^.

% C, 53.20;% H, 6.42;% N, 7.04.

B. Polymers (trimethylsilyl) - 3,6‘sdi-N-benzyls oxyaarbonylkanamycin · A

A mixture of 1.5 g (2 mmol) of 3,6‘-di-N-carbobenzocycloamycin A from Step A and 1.29 g (8 mmol) of hexamethyldisilazane in 15 ml of anhydrous acetonitrile was heated at reflux for 16 hours. The clear solution was then evaporated to dryness in vacuo and the residue was dissolved in 20 ml of anhydrous diethyl ketone. The solution was used directly in the next step.

C. Acylation of poles (trimethklsilyl) -3,6‘-di-N-

-benzocloxycarbborankanamycir A using an eaimolar amount of acylating agent

700 mg (2 mmol) of L- (-) -. Alpha.-benzocyclocarbonylamino-.alpha.-hydroxybutyric acid N-hydroxy-succinimidate ester was added to the solution of step B with stirring. The mixture is stirred at room temperature for 19 hours, then 8 ml of water and 35 ml of tetrahydrofuran are added, the pH is adjusted to 3 'by addition of aqueous hydrochloric acid solution, then the mixture is stirred for another 30 hours and then evaporated to dryness. vacuum. The residue is dissolved in a mixture of 30 ml. water, 40 ml of methanol, 10 ml of n-butanol and 40 ml of tetrahydrofuran and then hydrogenated overnight in the presence of 500 mg of 10% palladium on activated carbon catalyst. The catalyst is removed

The filtrate was evaporated in vacuo and lyophilized to give 1.7 g of crude BB-K8. The amorphous powder is dissolved in water, a solution. The solution was adjusted to pH 4 by addition of aqueous hydrochloric acid solution and then chromatographed on a resin column (Amberlite CG-50) in NH4 ⁺ -cycle. The column is washed sequentially with water, 0.1 N ammonium hydroxide, 0.3 N ammonium hydroxide, 0.5 N ammonium hydroxide and 2 N ammonium hydroxide, collecting fractions of. 10 ml and these fractions were monitored by thin-layer chromatography on silica (Merek Silica Gel 60 F-254) using chloroform / methanol / 28% ammonium hydroxide / water 1: 4: 2: 1 as the solvent. They are combined, evaporated and lyophilized again. The BB-K8 fraction and the kanamycin A fraction were tested using K. pneumoniae A20680 and B. subtilis PCI 129.

NHdOH fraction (N) Weight Yield number (mg) (%)

Product

Rf

32 to 39 0.1, 0.3 102 partially deblocked 40 to 46 0.3 Γ74 ^η 7 ^C kanamycin A 0.42 47 to 59 0.3, 0.5 106 unidentified 0.33 60 to 78 0.5 816 ^p 67 ^c BB-K8 0.18 89 to 95 2 70 5 diacylkanamycin AA 0.05 (a) 408 (g / mig (b) 956 fuZm-g

(c) according to the biological test

Hydrogenolysis of the partially deprotected product with palladium on the active. charcoal followed by isolation on a resin column (Amberlite CG-50) gave an additional 30 mg of BB-K8. (Yield 2%). The total yield of BB-K8 was as follows. increase to 69% (846 mg).

D. Acylation of poly (trimethylsilyl) -3,6‘-d-N-benzyloxycarbonylcanamycin A using 1.2 equivalents of acylating agent

Step C was repeated except that a 20% excess of acylating agent was used. The results are shown in the following table.

Fractions NH? OH Mass Yield Product Value number (N) (mg) (%) Rf

28 to 29 0.2 107 partially deblocked 30 to 41 0.2 157 unidentified 0.35 42 to 52 0.3 120 unidentified 0.30 53 to 81 0.3, 0.5 750 ³ 60 ^p BB-K8 0.14 94 to 116 1, 2 147 11 diacylkanamycin A 0.05 a) 933 .mu.gmg

(b) according to the biological test

Hydrogenolysis of the partially deprotected product with palladium on charcoal followed by isolation on a resin column (Amberlite CG-50) afforded an additional 21 mg of BB-K8 (2% yield). The total yield of BB-K8 will thus increase to 62 why. (771 mg).

E. Poly (trimethylsilyl) 3,6‘-di-N-banzyloxycarbonylkanamycin A acylation using 1.5 equivalents of acylating agent

Step C was repeated except that an excess of acylating reagent was used. The results are shown in the following table.

Fraction number NH4OH (N) Mass (mg) Yield (%) Product Rf value (dual development) 29 to 39 0.2 293 partially deblotted 40 to 47 0.3 95 ^a 6 ^C kanamycin A 0.67 65 to 80 0.5 582 ^p 29 ^c BB-K8 0.33 100 to 130 1, 2 543 26 diacylkanamycin A 0.09

(a) 860 (g / ^m

(b) 880. (zggmg

(c) according to the biological test

OBJECT OF THE INVENTION

Claims (8)

A process for the preparation of 1-N- (ω-amino-α-hydroxyalkanoyl) kanamycin A and B of formula I wherein: R is OH or amino, and n is an integer from 0 to 2 as well as non-toxic, pharmaceutically acceptable acid addition salts thereof, characterized by. by acylating the polysilylated kanamycin A or B or the polysilylated kanamycin A or B, containing a different protecting group from the silyl group on the 6'-amino group, provided that the polysilylated kanamycin A or B used as the starting material. material contains. an average of 4 to 8 silyl groups per molecule, and a polysilylated kanamycin A or B used as starting material and containing a 6'-amino protecting group different from the silyl group contains an average number of silyl groups in the 3 to 7 molecule, as acylation an acid derivative of the formula II is used B \ _B-N-CH2 - (b-CH 2 CH-COOH ·, OH (II), wherein n is an integer from 0 to 2, B is an amino protecting group or a hydrogen atom, provided that at least one of these symbols is an amino protecting group, namely o-nitrophenylthio, O CH. O o ^ 11 ¹ 3 II no CHgC-CH-C-O-C-YJCC-o II X C-CH 2 O-C-A or the symbols B together form a group O II II where R ¹ and R ^{2 are} identical or different and denote hydrogen, fluorine, chlorine, bromine, N02 »OH, alkyl or alkoxy of 1 to 6 carbon atoms, X represents a chlorine, bromine, fluorine or iodine atom; Y represents a hydrogen, chlorine, bromine, fluorine or iodine atom in a substantially anhydrous organic solvent followed by removal of all protecting groups, whereupon the resulting product is optionally converted to its non-toxic, pharmaceutically acceptable acid addition salt. 2. A process according to claim 1, wherein the acylating derivative of the acid of formula (II) is an active ester or mixed anhydride. 3. The process according to claim 1, wherein the acylating derivative of the acid of the formula II is an active N-hydroxysuccinimide ester, N-hydroxy-5-norbornene-2,3-dicarboximide ester or N-hydroxyphthalimide ester. 4. A process according to claim 3, wherein the acid acylation derivative of the formula (II) in which V is a benzyloxycarbonyl group. 5. The process of claim 1, wherein the acid acylation derivative is a mixed anhydride with pivalic acid, benzoic acid, isobutylcarboxylic acid or benzylcarboxylic acid. 6. The process according to claim 1, wherein the starting material is a polysilylated kanamycin A or B which contains a benzyloxycarbonyl group on the 6 &apos; -amino group as a protective group. 7. The process of claims 1 to 6, wherein the starting material is polysilylated kanamycin A or B, wherein the silyl groups are trimethylsilyl groups. 8. A process for the preparation of 1-N- [L- (-) -. Alpha.-amino-.alpha.-hydroxyalkanoyl] kanamycin A according to claims 1-7, wherein the acid acylation derivative is a compound of the formula: