CH618986A5 - Process for the preparation of aminoglycoside antibiotics. - Google Patents

Description

The invention relates to a process for the preparation of 5-epi derivatives of 4,6-di-0- (aminoglycosyl) -2-de-oxystreptamines which contain the gentamicins, sisomicin, verdamicin, tobramycin, the kanamycins, antibiotics G- 418,6640B, 6640D, JI-20A, JI-20B and G-52 and their 1-N-alkyl derivatives.

Broad-spectrum antibiotics are known in the literature, which can be classified chemically as 4,6-di-0- (aminoglycosyl) -1,3-diaminocyclitole. Valuable antibiotic active agents are those in which the aminocyclitol is 2-deoxystreptamine. Within the group of 4,6-di-0- (aminoglycosyl) -2-deoxystrepta-mines, those compounds in which the group in position 6 is a garosaminyl group are particularly preferred. The 4-0-aminoglycosyl-6-0-garosaminyl-2-deoxystreptamines include antibiotics such as Gentamicins B, Bi, Ci, Cla, C2, C2a, C2b, and X2; Verdamicin, Sisomicin, antibiotic G418, antibiotic G-52, antibiotic JI-20A and antibiotic JI-20B.

The 1-N-alkyl derivatives of the aforementioned 4,6-di-0- (aminoglycosyl) -1,3-diaminocyclitols are also known in the literature, which generally have a broadband spectrum and are particularly active against those microorganisms which are active are resistant to the 1-N-unsubstituted antibiotic.

The pseudotrisaccarides, produced according to the method of the invention, are derivatives of 4,6-di-0- (aminoglycosyl) -2-deoxystreptamine gentamicin A, gentamicin B, gentamicin Bi, gentamicin Ci, gentamicin C) a, gentamicin C2, gentamicin

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C2a, gentamicin C2b, gentamicin X2, tobramycin, verdamicin, kanamycin A, kanamycin B, 3 ', 4'-dideoxykanamycin B, antibiotic G-52, antibiotic 6640B, antibiotic 6640D, antibiotic G418, antibiotic JI-20A, antibiotic JI-20B and sisomicin, wherein the 2-deoxystreptamine group is replaced by a 1,3-diaminocyclitol of the formula I nh2

is, and in the formula R is hydrogen or the group -CH2Y, where Y is hydrogen, alkyl, alkenyl, cycloalkyl, cycloalkyl-alkyl, hydroxyalkyl, aminoalkyl, N-alkylaminoalkyl, amino-hydroxyalkyl, N-alkylaminohydroxyalkyl, phenyl, benzyl or tolyl means and the aliphatic radicals have up to seven carbon atoms and, if substituted by amino and hydroxy, have these substituents on different carbon atoms and X is hydroxy, and their pharmaceutically acceptable acid addition salts.

Particularly valuable antibacterial compounds are derivatives of 4,6-di-0- (aminoglycosyl> 2-deoxystreptamine, in which the aminoglycoside part in position 6 is garosaminyl.

Typical derivatives of 4-0-aminoglycosyl-6-0-garosaminyl-2-deoxystreptamines are derivatives of Gentamicin B, Gentamicin Bi, Gentamicin Ci, Gentamicin Qa, Gentamicin C2, Gentamicin C2a> Gentamicin C2b, Gentamicin X2, Sisomicin, Verdami-5 a, antibiotic G418, antibiotic JI-20A, antibiotic JI-20B and antibiotic G-52, which compounds are defined by the following formula X:

30th

and wherein X and R are as defined above for formula I and AGi represents one of the following aminoglycosyls:

ch nh 6'j 2 2 5 / o

/ \ (in derivatives of "^ / oH Gentamicin B)

H0

NH

çh.

oh ch3NH ~

"Ti

-O

(Gentamicin C ^)

_.h ch3 2

nh:

h "0,

(Gentamicin C ^)

(Gentamicin B ^)

(Gentamicin C ^)

nh,

ch.

H

-nh,

(Gentamicin C)

nh,

nh,

618986

4th

; h2nhch3

(Gentamicin C ^)

ch2oh

O.

HO

(Gentamicin X ^)

nh ch2nhj

/

~ 0.

(Sisomicin)

H-

ch3 ^ 2

NH "

/

(Verdamicin)

nh. £

ch2nh2

Oh

(Antibiotic ji-20a)

HO

nh.

ch.

H-

nh,

M

Oh

(Antibiotic ji-20b)

ho nh,

ch2nhch3

ch.

h0-

H

(Antibiotic G-52)

Oh

(in derivatives of antibiotic G-418)

ho nh

NH,

2 2

Other derivatives of 4,6-di-0- (aminoglycosyl) -2-deoxy-streptamines are derivatives of tobramycin, kanamycin A, kanamycin B and 3 ', 4'-dideoxykanamycin B of the formula XI:

NH "

AG,

NHR

O.

ch2oh nh,

Ho Oh

XI

wherein X and R are as defined for formula I and AG2 represents one of the following aminoglycosyls:

ch2nh2

CH2NH2

-0

(in derivatives of tobramycin)

HO

OH

(Kanamycin A)

HO

NH,

OH

ch2nh2

HO

(Kanamycin B)

ch2nh2

(in derivatives of

'3', 4 '-di - deoxykanamycin B);

NH,

Derivatives of antibiotic 66-40D of formula XII:

ch2NH2

NH.

XII

OH

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wherein X and R are as defined above for Formula I; and derivatives of gentamicin A and antibiotic 66-40B of the formula XIII:

OH

wherein X and R are as defined above for Formula I and AG3 is an aminoglycosyl having one of the following formulas

CH2OH 9H2m2

(in derivatives // \ (in derivatives of gentamicin \ of antibiotic A), and \ / 66-40b).

The derivatives of 4,6-di-0- (aminoglycosyl) -2-deoxystrepta- and cyclopentylethyl; aromatic groups such as o-, m-, p-Me-mine, which are defined by formula I and by formulas X-XIII thylbenzyl; straight and branched by hydroxy are white, amorphous powders. chain alkyl groups such as ß-hydroxyethyl, e-hydroxypentyl,

The pharmaceutically acceptable acid addition salts of ß-hydroxy-y-methylbutyl, ß-hydroxy-ß-methylpropyl,

the derivatives of 4,6-di-0- (aminoglycosyl> 2-deoxystrepta- 45 ô-hydroxybutyl, ß-hydroxypropyl, y-hydroxypropyl, co-hydro-

mines are produced by known methods, for example xyoctyl, ß-hydroxy-S-pentenyl; substituted by amino by neutralizing the free base with the appropriate straight and branched chain alkyl groups such as e-aminopentyl,

Acids up to a pH of approximately 5. Suitable acids are ß-aminopropyl, γ-aminopropyl, 5-aminobutyl, ß-amino-y-Me-

e.g. B. hydrogen chloride, sulfur, phosphorus and Bromwasser- thylbutyl and ra-aminooctyl and their mono-N-alkylated deri-

substance acid. The acid addition salts are usually white derivatives such as N-methyl, N-ethyl and N-propyl derivatives, for example

Solids that are soluble in water and in most polar and white e-methylammopentyl, ß-methylaminopropyl, ß-ethylami-

non-polar solvents are insoluble. The compounds denopropyl, 5-methylaminobutyl, β-methylamino-y-methylbutyl ser invention, as defined by formulas X-XIII, and insbe- and o-methylaminobutyl; substituted by amino and hydroxy

especially those in which the 6-0-aminoglycosyl is 6-O-garosaminylated straight and branched chain alkyl groups, as is, and the non-toxic, pharmaceutically acceptable acid 55β-hydroxy-e-aminopentyl, y-hydroxy-y-methyl - 8-aminobutyl,

Readition salts generally have antibacterial activity S-β-hydroxy-8-aminobutyl, S-ß-hydroxy-y-aminopropyl and with a broad spectrum and have a more advantageous spectrum ß-hydroxy-ß-methyl-y-aminopropyl; and their mono-N-alky-

as the underivatized antibiotics. lated derivatives such as ß-hydroxy-e-methylaminopentyl,

The substituent -CH2Y in the compounds includes y-hydroxy- y-methyl- 8-methylaminobutyl, ß-hydroxy- s-methyl-

straight and branched chain alkyl groups, such as ethyl, n-propyl, m laminobutyl, ß-hydroxy-y-ethylaminopropyl and ß-hydroxy-ß-

n-butyl, ß-methylpropyl, n-pentyl, ß-methylbutyl, y-methylbutyl methyl-y-methylaminopropyl.

and ß, ß-dimethylpropyl; n-Hexyl, ô-methylpentyl, ß-ethylbutyl, In a preferred group of compounds, R in y-ethylbutyl, n-heptyl, e-methylheptyl, ß-ethylpentyl, y-ethyl- of the formula I is hydrogen or alkyl with up to four coal

pentyl, ö-ethylpentyl, y-propylbutyl, n-octyl, iso-octyl, ß-ethyl atoms (preferably 2-4 carbon atoms), where hexyl, 8-ethylhexyl, e-ethylhexyl, ß-propylpentyl, y-propylpene - b5 hydrogen and ethyl and also propyl particularly preferably tyl; Alkenyl groups such as ß-propenyl, ß-methylpropenyl, ß-butene are.

nyl, ß-methyl-ß-butenyl, ß-ethyl-ß-hexenyl; cyclic groups, the process according to the invention is characterized by

such as cyclopropylmethyl, cyclopentylmethyl, cyclohexylmethyl, that is a derivative of any of the above

618986

4,6-Di-0- {aminoglycosyl> 2-deoxystreptamine, in which the 2-deoxystreptamine part is replaced by 1,3-diaminocyclitol of the formula II

NH.

NHR

, (»)

in which R has the above meaning and X 'represents unsubstituted or substituted hydrocarbon sulfonyloxy, and the hydroxyl and amino groups in the 4,6-di-0- (aminoglycosyl) -2-deoxystreptamine derivative are protected by groups which prevent reductive saponification or basic or weakly acidic hydrolysis, is treated with dimethylformamide at temperatures in the range from 80 to 155 ° C, the protective groups in the product obtained are removed and the derivative is isolated as such or as a pharmaceutically acceptable acid addition salt.

The Per-N-protected-per-O-protected-5-O-hydrocar-bonsulfonyl-4,6-di-0- (aminoglycosyl) -2-deoxystreptamines, which are starting materials for the process of this invention, are derived from known ones unprotected 4,6-di-O- (aminoglycosyl) -2-deoxy-streptamines including 4-O-aminoglycosyl-6-Ò-garosaminyl-2-deoxystreptamine antibiotics, e.g. B. Gentamicin B, Gentamicin Bi, Gentamicin Ci, Gentamicin Cia, Gentamicin C2, Gentamicin C2a, Gentamicin C2b, Gentamicin X2, Sisomicin, Verdamicin, Antibiotic G418, Antibiotic JI-20A, Antibiotic JI-20B and Antibiotic G-52, and 4 , 6-di-0- (aminoglycosyl) -2-deoxystreptamines, e.g. B. Gentamicin A, Tobramycin, Antibiotic 6640B, Antibiotic 6640D, Kanamycin A, Kanamycin B and 3 ', 4'-Dideoxykanamycin B,

from.

The 4,6-di-0- (aminoglycosyl) -2-deoxystrepta-min antibiotics mentioned are known. Of the gentamicins, the starting compound named Gentamicin X2 is also known as Gentamicin X. The starting compound named Gentamicin C2b with the structural formula shown is referred to in some prior publications as Gentamicin C2a.

When the 1-N-CH2Y derivatives of this invention are prepared via the corresponding starting materials, the IN-CH2Y-Per-N-protected-per-0-protected-5-0-hydrocarbonsulfonyl4,6-di-0- ( aminoglycosyl) -2-deoxystrep-tamine precursors also from the 1-N-CH2Y derivatives of the aforementioned 1 -N-unsubstituted4,6-di-0- (aminoglycosyl) -2-deoxystreptamine. These connections are known.

When the Per-N-protected-per-O-protected-5-O-hydrocarbonsulfonyl4,6-di-0- {aminoglycosyl) -2-deoxystreptamine compounds are prepared, the amino groups are first formed by the formation of amides, protected from reductive saponification or basic hydrolysis. For the process of this invention, it is preferred to remove the amino groups by forming N-benzyloxycycarbonyl derivatives (e.g., l, 3,6 ', 3 "tetra-N-benzyloxycarbonylgenamicin B and l, 3,2 ', 6', 3 "-Penta-N-benzyloxycarbonylgentami-cin Ci) to protect.

The per-N-protected aminoglyco-sides thus formed, which have a garosaminyloxy radical on the C-6 atom, are then treated with an alkali metal hydride, usually sodium hydride in dimethylformamide, whereby the

3 "-N-hydrocarbonyloxycarbonyl protecting group with the 4" hydroxy function is cyclized to form an oxazolidinone derivative, i. H. to form a 3 ", 4" -N, 0-carbonyl derivative. In amino glycosides, in which other amino groups are adjacent to a hydro-5 xyl group (e.g. the 6'-amino and 4'-hydroxyl in gentamicin B), other N.O-carbonyl derivatives are formed. In this way, after reaction of 1,3,2 ', 6', 3 "-penta-N-benzyloxycarbonylgentamicin Ci and 1,3,6 ', 3" -tetra-N -benzyloxycarbonylgentamicin B with sodium hydride in 10 dimethylformamide oxazolidinone Derivatives, i.e. H. 1,3,2 ', 6'-tetra-N-benzyloxycarbonyl-3 ", 4" -N.O-carbonylgentamicin Ci and l, 3-di-N-benzyloxycarbonyl-6', 4 '; 3 ", 4" -di-N, 0-carbonylgenta-micin B.

If amino groups are protected by bactericides 15 which cannot form a 3 ", 4" -N, 0-carbonyl derivative (e.g. intermediates which do not have a 6-O-garosaminyl substituent, such as gentamicin A and 3 ', 4'-di-deoxykanamycin B), the amino groups are expediently by lower alkanoyl groups, for. As acetyl and propionyl, protected, 20 the bactericide is treated with the corresponding acid anhydride in ethanol, whereby the corresponding per-N-lower alkanoyl aminoglycoside is produced. In this way, z. B. by treatment of G entamicin A with acetic anhydride in ethanol Per-N-acetylgentamicin A. 25 Although any amino protective group can be used, particularly suitable amino protective groups for the starting compounds of the process according to the invention include lower alkoxycarbonyls (preferably with up to 8 carbon atoms, e.g. methoxycarbonyl, ethoxycarbonyl, n-propyloxycarbonyl, 30-iso-propyloxycarbonyl, n-butyloxycarbonyl, tert-butoxycarbonyl, octyloxycarebonyl and the like), substituted benzyloxycarbonyl (including o-, m- and p-methoxybenzyloxycarbonyl and the like), and preferably benzyloxycarbonyl. Lower alkanoyles with preferably up to 8 carbon atoms (e.g. 35 acetyl, propionyl, valeryl, caprylyl) are also suitable amino protecting groups, in particular for compounds which are derived from bactericides which do not contain 3 ", 4" -N, 0- Can form a carbonyl derivative (e.g. compounds that do not have a 6-0-Garosa-minyl substituent, such as derivatives of Gentamicin A 40 and Kanamycin).

The aforementioned amino protecting groups can be removed by treatment with base (for example with sodium hydroxide) or, in the case of benzyloxycarbonyl, by known reductive saponification methods. Benzyloxycarbonyl is preferred as the amino protecting group for the starting compound in this process, since in amino glycosides having a hydroxyl function adjacent to an amino function, e.g. B. in positions 3 "and 4" in the 6-O-garosaminyl radical of the aminoglycosides of the formula X, the N-benzyloxycarbonyl derivative (z. B. the 3 "-N-50 benzyloxycarbonyl derivatives of the compounds of the formula X) - when subjected to basic conditions (e.g. with sodium hydride in dimethylformamide), an oxazolidinone with the adjacent hydroxyl function (e.g. a 3 ", 4" -N, 0-carbonyl derivative of compounds of formula X) with simultaneous elimination of benzyl alcohol. Similarly, N-alkoxycarbonyl derivatives form oxazolidinones with an adjacent hydroxyl function. If a starting compound has a 1-N-CH2Y substituent which has a hydroxyl group in a- or has a β-position to an amino protective group bo which is benzyloxycarbonyl or alkoxycarbonyl, the hydroxyl group forms together with the protective group an oxazolidinone or tetrahydro-1,3-oxazin-2-one.

Hydroxyl functions in the starting compounds of this process are expediently protected by O-acyl residues 65 of hydrocarboncarboxy acids, preferably having up to 8 C atoms, or by O-hydrocarbonylidene residues of ketones and aldehydes, preferably having up to 8 C atoms , with ketals or acetals including cyclic

618986

Ketals and acetals are formed, although any other hydroxy protecting group can be used to protect the hydroxyl functions.

In general, adjacent hydroxyl groups in the aminoglycoside precursors of the starting compounds of this process are expediently protected by cyclic ketals or acetals. Adjacent hydroxyl groups are understood to be vicinal and non-vicinal hydroxyl groups which are arranged such that they form a cyclic ketal or cyclic acetal function together with ketones or aldehydes or their derivatives. Examples of such adjacent hydroxyl groups are the 2 ', 3'-hydroxyl groups in gentamicins B and Bi and in kanamycin A (which form 2', 3'-0-hydrocarbonylidene derivatives), the ^^ '- hydroxyl groups in gentamicins A and X2 and in antibiotic G418 (which form 4 ', 6'-0-hydrocarbonylidene derivatives), the 3', 4'-hydroxyl groups in antibiotics JI-20A and JI-20B and in kanamycin B (which 3 ', 4'-0- Form hydrocarbonylidene derivatives) and the 4 ", 6" hydroxyl groups in tobramycin, kanamycin A and B and 3 ', 4'-dideoxykanamycin B (which form 4 ", 6" -0-hydrocarbonylidene derivatives).

The cyclic ketal and acetal derivatives of the adjacent hydroxyl groups include O-alkylidene (e.g. O-isopropylidene), O-cycloalkylidene (e.g. O-cyclohexylidene) and O-aryl-alkylidene (e.g. O-benzylidene) derivatives, all of which can be removed by treatment with dilute, aqueous, weak acid (e.g. with 50 to 80% acetic acid). The type of hydrocarbon or substituted hydrocarbon “ylidene” radicals of the cyclic ketals and acetals is immaterial since they only act as blocking groups and do not enter the process according to the invention and are subsequently removed, so that the free hydroxyls are in their original form Shape to be restored.

In the starting compounds of the process of this invention, isolated hydroxyl groups other than the 5-hydroxyl group, e.g. B. the 2 "-hydroxy in all aminoglycoside precursors, the 4'-hydroxy in tobramycin and the 4" -hydroxy in antibiotic 6640B and in gentamicin A as well as other hydroxyl groups which are not protected by cyclic ketal or acetal functions ( e.g. the 3'-hydroxy in gentamicin A and the 2'- and 4'-hydroxyl in kanamycin A) suitably protected by hydrocarboncarbonyl residues,

the hydrocarbon preferably having up to 8 carbon atoms. Hydrocarboncarbonyl residues which can be used are acyl residues which are derived from lower alkanoic acids having up to 8 carbon atoms, such as acetyl, propionyl, n-butyryl, valeryl and caprylyl, and acyl residues which are derived from aralkanoic acids, e.g. As phenylacetyl, and acyl residues derived from arylcar-boxylic acids, for. B. o-, m- and p-toluoyl, mesitoyl and preferably benzoyl.

Usually, after the introduction of the amino protecting groups, adjacent hydroxyl groups are protected which, after treatment with a ketone or aldehyde or a derivative thereof in dimethylformamide in the presence of catalytic amounts of a strong acid, e.g. B. p-toluenesulfonic acid, using known methods form a ketal or acetal group. In this way, from the intermediate gentamicin B mentioned after treatment with 1,1-dimethoxycyclohexane in dimethylformamide in the presence of p-toluenesulfonic acid, a ketal at the 2'- and 3'-hydroxylene, ie. H. 1,3-di-N-benzyloxycarbonyl-2 ', 3'-0-cyclohexylidene-6', 4 '; 3 ", 4" -di-NO-carbonylgentamicin B. Finally, each isolated hydroxyl function (except the 5-hydroxyl group) which remains in the partially protected aminoglycoside derivatives is treated by treatment with an acid chloride of hydrocarboncarboxylic acid in a tertiary amine ( preferably pyridine), the molar amount of the acid halide being based on the number of hydroxyl groups to be esterified, converted to the corresponding hydrocarboncarbonyl derivatives. If only one hydroxyl group remains in the molecule in addition to the 5-hydroxy (e.g. the 2 '' hydroxy), an equivalent amount of the acid halide to the molar amount of the aminoglycoside is used; if two remaining hydroxyl groups are to be protected, two molar equivalents of acid halide are used per mole of aminoglycoside. Acid halides of hydrocarboxyl acids with up to 8 carbon atoms are preferably used, such as, for. B. acid chlorides of lower alkanoic acids, e.g. B. acetic, propionic, valeric and caprylic acid, of aralkanoic acids, e.g. B. phenylacetic acid, and of arylcarboxylic acids, e.g. B. toluic and preferably benzoic acid. In this way, the corresponding 2 ″ -O-benzoyl derivative, that is to say, is obtained from the aforementioned intermediates of gentamicin Ci and B after treatment with equimolar amounts of benzoyl chloride in pyridine. H. 1,3,2 ', 6'-tetra-N-benzyloxycarbonyl-2' '-O-benzoyl-3' ', 4' '-N, 0-carbo-nylgentamicin Ci and 1,3-di-N-benzyloxycarbonyl -2 ', 3'-0-cyclohexylidene-6', 4 '; 3 ", 4" di-N, 0-carbonyl-2 "-O-benzoylgen-tamicin B, both of which give the corresponding 5-O-methanesulphonyl derivatives after treatment with methanesulfonyl chloride in triethylamine.

After protecting the amino group using N-benzyloxycarbonyl derivatives and the hydroxyl groups adjacent to the protected amino groups via N, 0-carbonyl derivatives, all other hydroxyl groups with the exception of the 5-hydroxyl group can alternatively be protected by conversion to a hydrocarbonyl group without neighboring hydroxyl groups are initially protected by acetal or ketal groups. In this way, 1,3-di-N-benzyloxycarbonyl-6 ', 4'; 3 ", 4" -di-N, 0-carbonylgentamicin B after reaction with three molar equivalents of benzoyl chloride in pyridine the corresponding 2 ', 3', 2 "-Tri-0-benzoate derivative, which after reaction with methanesulfonyl chloride in pyridine to a Compound which can be used according to the invention, ie 1,3-Bi-N-benzyloxycarbonyl-5-0-methanesulfonyl-2 ', 3', 2 "-tri-0-ben-zoyl-6 ', 4'; 3 ", 3" -di-N, 0-carbonylgentamicin B, is converted.

The reaction according to the invention is carried out in dimethylformamide alone or preferably also in the presence of a tetraalkylammonium alkanoate. Treatment of a 5-O-hydrocarbonsulfonyl intermediate with dimethylformamide alone is mostly carried out at the reflux temperature (i.e. about 155 ° C), since in this case the reaction rate is faster than when the reaction is carried out at lower temperatures; however, when the reaction is carried out in the presence of a tetraalkylammonium alkanoate, the reaction proceeds well at lower temperatures (e.g. 100-140 ° C) in order to obtain good yields of a purer product. Tetra-n-butylammonium acetate is usually chosen, but other tetraalkylammonium alkanoates, e.g. B. tetraethylammonium acetate, tetramethylammonium acetate, tetraethylammonium formate, tetra-n-butylammonium formate and the like can be used. The molar amount of tetraalkylammonium alkanoate per mole of aminoglycoside is usually between about 1.5 and about 5 moles.

The intermediates prepared after reaction of the N-protected-O-protected-5-O-hydrocarbonsulfonyl (or substituted hydrocarbonylsulfonyl) 4,6-di-0- (aminoglycosyl) -2-deoxystreptamine with dimethylformamide give a 5-epi after hydrolysis -Connection.

When tetra-n-butylammonium acetate is used in conjunction with dimethylformamide, the intermediate produced is the corresponding more N-protected-5-epi-O-acetyl derivative, which gives a 5-epi compound after hydrolysis.

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Protective groups accessible to reductive saponification, benzyloxycarbonyl-5-0-methanesulfonyl-2 "-0-benzoyl-3", 4 ", are most preferred when carrying out the process, NO-carbonylsisomicin, in dimethylformamide, to which they are easily exposed after epimerization added to the 5-position by reduc-tetra-n-butylammonium acetate, 16 h tive methods can be removed at 120 ° C. Other protective groups are heated and the solution is evaporated, but the corresponding pen will remain after the reduction stage, e.g. 5 5-epiacetyl derivative, e.g. 1, 3,2 ', 6'-tetra-N-benzyloxycarbonyl-

N, 0-carbonyl groups, which are removed by treatment with aqueous 5-epi-0-acetyl-2 "-0-benzoyl-3", 4 "-N, 0-carbonylsisomicin and ger base at elevated temperatures 1-N-ethyl derivative is obtained, which furthermore it is necessary to connect acetals or ketals by acid hy after treatment with aqueous potassium hydroxide.

remove drolysis. neutralization, isolation and cleaning by means of

If protective groups which a reductive saponification io matographic methods a 5-epi compound, z. B. 5-Epi accessible, results from the bisisomicin and 1-N-ethyl-5-episisomicin produced in this process.

products are removed, the reduction with water, acetal or ketal protective groups in the intermediate product in the presence of a catalyst is preferred if the products are removed after the N protective groups have been removed by 5-epi-4,6-di-0 - (Aminoglycosyl) -2-deoxystreptamine-N- treatment with dilute acid, e.g. B. dilute mineral acid-protected-O-protected-intermediates not saturated 15 ren, dilute trifluoroacetic acid or usually with, e.g. B. when the intermediates by treating thin alkanoic acids, e.g. B. acetic acid removed. 0- and N-protected derivatives of gentamicins A, B, Bi, Ci, when carbobenzyloxy protecting groups from a Per-N-Cja, C2, C2a, C2b and X2, from tobramycin, Kanamycins A and B, protected-per-O-protected -Amino-glycoside intermediate pro-3 ', 4'-dideoxykanamycin B and antibiotics G418, JI-20A and ducts with a double bond (e.g. the intermediate product JI-20B obtained with dimethylformamide. If protection- 20 differs from l, 3,2 ', 6'-tetra-N-benzyloxycarbonyl-2 "-0-benzoyl groups, which are accessible to reductive saponification, from 3", 4 "-N, 0-carbonylverdamicin after treatment with dimethyl-N-protected -0-protected-5-epi-4,6-di-0- (aminoglycosyl) -2- formamide derivatives) by reaction with an alkali metal (e.g. deoxystreptamine intermediates, in which- potassium, lithium and preferably Sodium) is present in liquid double bonds, e.g. those that are removed from ammonia, the intermediate product sisomicin, verdamicin, antibiotic G-52, antibiotics 6640 B 25 usually in a mixed solvent, e.g. B. tetrahydrofuran and 6640D, the reduction is preferably by means of and liquid ammonia, to which the alkali metal (e.g. N atrium) of an alkali metal in liquid ammonia is added, is added, and the reaction mixture reduces the reduction of the double bond for a few hours prevent. touched. The ammonia is allowed to evaporate, all are removed. If the protective groups with remaining O and N protective groups (for example the 3 ", 4" -N, 0-carbo-hydrogen are removed in the presence of a catalyst, 30 nyl in an intermediate product) - and 2 "-0-benzoyl group) by adding water to the most commonly used catalyst is palladium, preferably a reaction mixture, whereby sodium hydroxide forms, and palladium on carbon. The hydrogenolysis of the protective groups heats up to higher temperatures (eg 100 ° C.) The cleaning is usually carried out at room temperature in lower alkanoic acids, the product obtained is carried out by means of chromatographic, preferably acetic acid, although other methods can be used, an antibacterial 5-epi-aminogly solvent, for example lower alkanols, being used. 35 coside, e.g. 5-epiverdamicin, is obtained.

nen. The hydrogenation is continued until no further. The protective groups can also be determined from the drop which can be seen after treatment in the hydrogen pressure, and an N- and 0-protected-5-0-hydrocarbonsulfonyl4,6-di-0-

the 5-Epi4,6-di-0- (aminoglycosyl) -2-deoxystreptamine is then (aminoglycosyl) -2-deoxystreptamine with dimethylformamide usually by removing the solvent, e.g. B. N- and O-protected intermediate obtained by distillation, and then the N- and 40 reaction thereof with base at elevated temperatures and

0-protected-5-epi-2-deoxystreptamine intermediate with treatment if acetals or ketals are present

Base treated and, if acetals or ketals are present, removed with aqueous acid.

which are, with aqueous acid to remove the remaining protection. The invention also relates to the use of the groups. Compound of the Forge Prepared by the Above Process In a typical embodiment of this process, 45 mei, where R is hydrogen, is used to prepare compounds - a 5-O-hydrocarbonsulfonyl-per-N-protected-per-O- of the formula I, wherein R represents the group -CH2Y, which protected intermediate, e.g. B. l, 3,2 ', 6'-tetra-N-benzyl- characterized in that the starting compound oxycarbonyl-5-0-methanesulfonyl-2 "-0-benzoyl-3", 4 "-N, 0 -car- with an aldehyde of the formula Y-CHO, in which Y the above bonylgentamicin Ci, dissolved in dimethylformamide and 18 h is significant in the presence of a hydrogen donor reduc reflux temperature, then the solution is a 50 and with optional intermediate protection, 1 residue containing a 5-epi-N-protected-O-protected- of amino groups in the compound of the formula I with Aus-4,6-di-0- (aminoglycosyl) -2-deoxystreptamine intermediate, would take the amino group in position 1, and each evaporated existing, which dissolved in acetic acid and at room temperature, the amino or hydroxyl group in the aldehyde of the formula temperature and 4 atm starting hydrogen pressure in the presence of Y-CHO and the compound obtained as such or one 30% palladium-carbon catalyst hydrogenates 55 as pharmaceutically acceptable Acid addition salt is isolated. If no further drop in hydrogen pressure is detected- This method, in which the 1-amino function is in bar, removes the catalyst by filtration, which removes a 1-N-unsubstituted derivative of a 4,6-di-0- {aminoglyco solvent Distillation removed in vacuo, the ent- Syl) -1,3-diaminocyclitol bactericide is selectively condensed with an aldehyde residue with 2N sodium hydroxide at elevated and simultaneously reduced in situ to a temperature (eg 100 ° C) , then neutralized with acetic acid eo 1N-CH2Y derivative of 4,6-di-0- (aminoglycosyl) -l, 3-diaminocyne and then using known clitol bactericide, is usually isolated and purified by room temperature methods , 5-Epigentamicin Ci ratur carried out in the presence of air, although it is obtained advantageously. in an inert atmosphere (e.g. argon or

According to another preferred embodiment, the nitrogen can be carried out.

This process uses a 5-O-hydrocarbonsulfonyl-per-N-65. The hydride-donating reducing agents include dialkyl-

protected-by-O-protected-intermediate, e.g. B. 1,3,2 ', 6' - aminoborane (e.g. dimethylaminoborane, diethylaminoborane

Tetra-N-benzyloxycarbonyl-5-0-methanesulfonyl-2 "-0-benzoyl- and preferably morpholinoborane), tetraalkylammoniumcy-

3 ", 4" -N, 0-carbonyl-sisomicin and l-N-ethyl-l, 3,2 ', 6'-tetra-N-anborohydride (e.g. tetrabutylammonium cyanoborohydride), alka-

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limetal borohydrides (e.g. sodium borohydride) and preferably alkali metal cyanoborohydrides (e.g. lithium cyanoborohydride and sodium cyanoborohydride).

This process is conveniently carried out in an inert solvent. Although anhydrous aprotic solvents can sometimes be used advantageously in these processes (e.g. tetrahydrofuran when using morpholine borane as the hydride donating reducing agent), this process is usually carried out in protic solvents, e.g. B. in a lower alkanol or preferably in water or in an aqueous, lower alkanol (z. B. aqueous methanol, aqueous ethanol), although other water-miscible mixed solvent systems can be used, e.g. B. aqueous dimethylformamide, aqueous hexamethylphosphoramide, aqueous tetrahydrofuran and aqueous ethylene glycol dimethyl ether.

The process is expediently carried out at a pH in the range from 1 to 11, preferably from 2 to 5, in particular from 2.5 to 3.5. The preferred acidic medium can be added by adding an organic acid, e.g. B. acetic acid, trifluoroacetic acid, or p-toluenesulfonic acid, or an inorganic acid, e.g. As hydrochloric acid, sulfuric acid, phosphoric acid or nitric acid to the aminocyclitol derivative can be obtained. This forms acid addition salts and it is usually very convenient to use the addition salts derived from sulfuric acid. Optimal results are obtained when all amino groups in the molecule are completely neutralized. It is usually convenient to add the required acid addition salt starting compound in situ by adding the desired acid (e.g. sulfuric acid) to a solution or suspension of the derivative of 4,6-di-0- (aminoglycosyl) -1,3 di-aminocyclitol in a protic solvent (e.g. water) until the pH of the solution is adjusted to the desired value.

In order to keep side reactions to a minimum when an amino aldehyde is used as a reagent, it is preferred to protect the amino function in the aldehyde, e.g. With an acyl blocking group such as acetamido, phthalimido or the like before carrying out this process and then removing the N-protecting group in the product obtained. It may also be advantageous to protect the hydroxyl group in hydroxyl-containing aldehydes when carrying out this process, but this is not generally necessary.

The following examples illustrate the invention:

Example 1 :

Per-N-benzyloxycarbonylaminoglycosides

A. l, 3,2 ', 6', 3 "-penta-N-benzyloxycarbonylgentamicin Qa

40 g Gentamicin Cia are dissolved in 200 ml methanol and 20 ml saturated sodium bicarbonate solution and the solution is cooled to 0 ° C. While the solution is being stirred, 88 ml of carbobenzyloxychloride are added dropwise over 2 hours and the reaction temperature is kept between 0 ° C and 5 ° C. The mixture is stirred overnight, allowing the reaction temperature to rise to room temperature. 500 ml of chloroform are added to the reaction mixture, which then separates into two layers. The organic phase is washed with 4x 100 ml of water and dried over 100 g of sodium sulfate. The organic phase is evaporated under vacuum at a temperature below 40 ° C. The crude product obtained is dissolved in 100 ml of chloroform and added dropwise to 250 ml of 75% hexane / ether. The precipitate obtained is filtered, washed with 100 ml of hexane and air-dried; 87 g (87%) l, 3.2 ', 6', 3 "-penta-N-benzyloxycarbonylgentamicin C) a are obtained, mp = 185-190 ° C, [a] g6 + 71.2 (CHsOH) , Infrared (IR) (KCl): 3300, 3500 cm -1, PMR (CDCL): 5 1.2 (C-Me), 3.0 (N-Me), 7.25 (aromatic H).

B. 1,3,2 ', 6', 3 '' penta-N-benzyloxycarbonylsisomicin

25 g sisomicin and 13 g sodium carbonate are dissolved in 625 ml 5 water. While the solution is being stirred, 100 ml of carbobenzyloxychloride are added at 25 ° C. The mixture is stirred for 16 h and then the solid is filtered off and washed with water. The solid is dried under vacuum, and then washed with hexane and air-dried; 62 g of 1,3,2 ', 6', 3 '' penta-N-benzyloxycarbonylsi-somicine are obtained. Mp = 165-173 ° C; [aß6 + 96.2 (CH30H), infrared (IR) = v max (CHCb), 3600.1720.15151515155050.695 cm "1; PMR Ô (CDCb) 1.03 (3H, broad singlet, 4 "-C-CH3), 3.02 (3H, broad singlet, 3" -N-CH3), 5.02 (10H, broad singlet -CHîCsHs), 3.28, 15 3.30 ppm. (25H, broad singlet -CHîCsHs).

Example 2:

Per-N-benzyloxycarbonyl-3 ", 4" -N, 0-carbonylaminoglycosides

A. 1,3,2 ', 6' -Tetra-N-benzyloxycarbonyl-3 ", 4" -N, 0-carbo-20 nylgentamicin Cia

A solution of 2 g of the product according to Example 1A in 50 ml of dry dimethylformamide is added to a stirred mixture of 60 mg of sodium hydride in 5 ml of dry dimethylformamide during one hour at room temperature under nitrogen. The reaction mixture is stirred for 2 hours and then the insolubles are filtered off. 100 ml of chloroform are added to the filtrate and the organic phase is washed with 3x50 ml of water. The organic phase is dried over 25 g of sodium sulfate and then evaporated under reduced pressure. The residue obtained is dissolved in 15 ml of chloroform and added dropwise to 75% hexane / ether (15 ml). The precipitate is filtered off and washed with 25 ml of hexane, 1.82 g (> 95%) of l, 3,2 ', 6'-tetra-N-benzyloxy-carbonyl-3 ", 4" -N, 0-carbonylgentamicin Cia can be obtained. 35 mp = 215 ° C (dec.) [A] ß6 + 63.4 infrared (IR) - (KCl) = 3300, 3500.1680.1545, PMR (CDCb) 5 1.28 (C-Me), 2.58 (N-Me), 7.25 (aromatic H).

B. l, 3,2 ', 6'-tetra-N-benzyloxycarbonyl-3 ", 4" -carbonylsisomi-40

250 mg of sodium hydride are added to a stirred solution of 5 g of the product according to Example 1B in 50 ml of dimethylformamide. The reaction mixture is stirred under argon for 2 hours at room temperature. It is filtered off and 2 ml of 45 glacial acetic acid are added to the filtrate. The filtrate is concentrated in vacuo and the residue is extracted with 200 ml of chloroform (purified by passing it through basic aluminum oxide). The chloroform extracts are washed with water and dried over sodium sulfate and evaporated to give 3.5 g of l, 3,2 ', 6'-tetra-N-benzyloxycarbonyl-3 ", 4" -N.O-carbonylsisomicin. Mp = 210-213 ° C; [a] g6 + 68.8 (c 0.22) infrared (IR) v max (Nujol) 3550.1760.1580, enr1 PMR 5 (CDCb) 1.34 (3H, singlet-4 "-CH3), 2 , 68 (3H, singlet-3 "-N-CH3), 5.04 (8H, broad singlet-CT ^ CsHs).

55

Example 3:

Per-N-benzyloxycarbonyl-2 "-0-hydrocarboncarbonyl-3", 4 "-N.O-carbonylaminoglycosides

A. 1,3,2 ', 6' -Tetr a-N-benzyloxycarbonyl-2 "-O-benzoyl-3", 4 "-60 N.O-carbonylgentamicin Cla

To a stirred solution of 10 gl, 3,2 ', 6'-tetra-N-ben-zyloxycarbonyl-3' ', 4' '-N, 0-carbonylgentamicin Ciain 50 ml dry pyridine for 10 to 15 min under a Nitrogen atmosphere 2 ml of benzoyl chloride added dropwise-65. The reaction mixture is stirred for Vi h and then the pyridine is removed on a rotary evaporator, the bath temperature being kept below 30 ° C. The light yellow oily residue is dissolved in 100 ml of chloroform. This organic

11

618 986

Phase is washed with 3 x 50 ml of water and then dried over 25 g of sodium sulfate. The chloroform is evaporated off under vacuum. The remaining yellow foam is triturated with a small volume of ether, 11.0 g (> 95%) 1,3,2 ', 6' -T etra-N-benzyloxycarbonyl-2 '' -O-benzoyl-3 ", 4 "-N, 0-carbonylgentamicin CIa can be obtained. Mp = 120-123 ° C [ctB6 + 73.8.

B. l, 3,2 ', 6'-tetra-N-benzyloxycarbonyl-2' '-O-benzoyl-3 ", 4" -N, 0-carbonylsisomicin

1.7 ml of benzoyl chloride are added to a stirred solution of 3 g of the product according to Example 2B in 20 ml of dry pyridine at 25 ° C. under an argon atmosphere for 10 minutes. The mixture is stirred at room temperature until all of the starting material has reacted (check by thin layer chromatography). The mixture is evaporated at room temperature under high vacuum; the solid residue was extracted with 100 ml of chloroform (which had previously been passed through basic aluminum oxide). The chloroform extracts are washed with 5% aqueous sodium bicarbonate and water and then dried over sodium sulfate. The solvent is evaporated to give 2.8 g of 1,3 ', 2', 6'-tetra-N-benzyloxycarbonyl-2 "-0-benzoyI-3", 4 "-N, 0-carbonyl-sisomicin. Mp = 157-160 ° C, [a] £ 6 + 86, (c 0.2) infrared (IR) v max (Nujol) 3325.1780.1680.1560 cm-1 PMR 8 (CDCb) 1.35 (4 "-C-CH3), 2.74 (3" -N-CH3), 5.03 (CHz-CeHs).

C. l, 3,2 ', 6'-tetra-N-benzyloxycarbonyl-2 "-0-acetyl-3" -N, 0-car-bonylgentamicin Qa

1. l, 3,2 ', 6', 3 "-penta-N-benzyloxycarbonyl-2" -0-acetylgen-tamicin Cla

1.4 ml of acetic anhydride are added dropwise to a stirred solution of 10 g of 1,3,2 ', 6', 3 "-penta-N-benzyloxycarbonylgentamicin Cla in 50 ml of dry pyridine over a period of 10 to 15 minutes under a nitrogen atmosphere. The reaction mixture is stirred for Vi h and then the pyridine is removed on a rotary evaporator, the bath temperature being kept below 30 ° C. The resulting residue is dissolved in 100 ml of acid-free chloroform. The organic solution is washed with 3x50 ml of water, dried over sodium sulfate and The residue formed is purified by trituration with small volumes of ether, 1,3,2 ', 6', 3 "-penta-N-benzyloxycarbonyl-2" -0-acetylgentamicin Cia is obtained.

2. l, 3,2 ', 6'-tetra-N-benzyloxycarbonyl-2 "-0-acetyl-3", 4 "-N, 0-carbonylgentamicin Cia

In a similar manner to that described in Example 2A, 1,3,2 ', 6', 3 "-penta-N-benzyloxycarbonyl-2" -O-acetylgentamicin Cla is treated with sodium hydride in dry dimethylformamide. The resulting product is isolated and purified as described in Example 2A, l, 3,2 ', 6'-tetra-N-benzyloxy-carbonyl-2 "-0-acetyl-3", 4 "-N, 0- carbonylgentamicin Qa is obtained.

D. l, 3,2 ', 6'-tetra-N-benzyloxycarbonyl-2 "-0-acetyl-3", 4 "-N, 0-carbonylsisomicin

1. In a similar manner as in Example 3C1. 1, 3,2 ', 6', 3 "-penta-N-benzyloxycarbonylisomicin is treated with acetic anhydride in pyridine. The resulting product is isolated and purified as described in Example 3C1., 1,3,2 ', 6 '-T etra-N-benzyloxycarbonyl-2 "-O-acetsisomicin is obtained.

2. In a manner similar to that described in Example 2B, 1,3,2 ', 6', 3 "-penta-N-jenzyloxycarbonyl-2" -O-acetylsisomicin is treated with sodium hydride in dimethylformamide. The product is isolated and purified as described in Example 2B to give l, 3,2 ', 6'-tetra-N-benzyloxycarbonyl-2 "-0-acetyl-3", 4 "-N, 0-carbonylsisomicin .

Example 4:

Per-N-benzyloxycarbonyl-2 "-0-hydrocarboncarbonyl-5-0-hydrocarbonsulfonyl-3", 4 "-N, 0-carbonylaminoglycosides

A. 1,3,2 ', 6' -T etra-N-benzyloxycarbonyl-5-0-methanesulfonyl-2 "-O-benzoyl-3", 4 "-N.O-carbonylgentamicin Q a

A solution of 1 gl, 3,2 ', 6'-tetra-N-benzyloxycarbonyl-2 "-0-benzoyl-3", 4 "-N, 0-carbonylgentamicin Cla in 5 ml of triethylamine and 15 ml of tetrahydrofuran cooled to 0 ° C. The solution is stirred and a solution of 1 ml of methanesulfonyl chloride in 5 ml of tetrahydrofuran is added to it over the course of 15 minutes, the reaction mixture is stirred for 2 hours at 0 ° C. The reaction mixture is dissolved in 25 ml of water and 25 ml of chloroform is poured in. The organic phase is washed with 2 × 15 ml of water and then the organic phase is dried over sodium sulfate. The chloroform is evaporated off and the resulting yellow foam is triturated with small amounts of ether, 1.2 g (> 95%). l, 3,2 ', 6'-tetra-N-benzyloxycar-bonyl-5-0-methanesulfonyl-2 "-0-benzoyl-3", 4 "-N, 0-carbonyl-gentamicin Cla can be obtained. Mp = 130 ° C [a] g5 + 53.4 (CHCh) PMR (CHCb) Ô 1.35 (C-Me), 2.74 (N-Me), 2.99 (OSO2CH3), 7.28 (4x Cbz and Benzoyl).

Similarly, 1, 3,2 ', 6'-tetra-N-benzyloxycarbo-nyl-2 "-0-acetyl-3", 4 "-N, 0-carbonylgentamicin, Qa in triethylamine and tetrahydrofuran with methanesulfonyl chloride treated, whereby l, 3,2 ', 6'-tetra-N-benzyloxycarbonyl-5-0-methansulfonyl-2 "-0-acetyl-3", 4 "-N, 0-carbonylgentamicin Qa is obtained.

B. l, 3,2 ', 6'-tetra-N-benzyloxycarbonyl-5-0-methanesulfonyl-2 "-0-benzoyl-3", 4 "-N, 0-carbonylsisomicin

2.5 g of the product according to Example 3B are dissolved in 15 ml of dry pyridine. The solution is cooled to 10 ° C and 4 ml of methanesulfonyl chloride are added over 10 minutes, the reaction mixture is left to stand overnight and then concentrated under vacuum at 25 ° C. The residue is extracted with 150 ml of acid-free chloroform. The chloroform extract is washed with water and dried over sodium sulfate. The chloroform is evaporated off, giving 2.4 gl, 3,2 ', 6'-tetra-N-benzyloxycarbonyl-5-0-methanesulfonyl-2 "-0-benzoyl-3", 4 "-N, 0 -carbonylsisomicin; mp = 84-88 ° C, [a6 + 21.3 (c 0.29) infrared (IR) v max (nujol) 3325.1750.1540 cm "1; PMR a (CDCb) 1.32 (4 "-C-CH3), 2.68 (3" -N-CH3), 3.04 0

(5.0 - ^ - ch3), 5.00 (-chzcéhs).

O

Example 5:

5-epigentamicin Ci

A. 2 g of 1,3,2 ', 6'-tetra-N-benzyloxycarbonyl-5-0-methanesulfonyl-2 "-O-benzoyl-3", 4 "-NO-carbonylgentamicin Ci are added to 15 ml of dimethylformamide, it is heated at reflux temperature for 18 h and then the solution is evaporated to a residue which contains N-protected-O-protected-intermediate product.

B. This residue is dissolved in acetic acid and 500 mg of a 30% palladium-charcoal catalyst are added and hydrogenated at room temperature starting from 4 atm hydrogen pressure. The catalyst is separated off by filtration and the filtrate is evaporated to a residue. The residue is dissolved in 25 ml of 5% sodium hydroxide and heated at 100 ° C for 4 h. The solution is cooled and passed through an IRC-50 (H + form) column, the resin column is washed well with water and then the product is eluted with 200 ml of ammonium hydroxide. The ammonium hydroxide eluate is concentrated to a residue which contains 5-epigentamicin Ci. The product is purified by chromatography on a silica gel column and eluting with the bottom layer of a chloroform / methanol / 15% ammonium hydroxide (2: 1: 1>

5

10th

15

20th

25th

30th

35

40

45

50

55

60

65

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Solvent system cleaned. The similar eluates, which are determined by thin layer chromatography, are collected and lyophilized to a residue, 5-epigentamicin Ci being obtained as the residue as a white solid; Mp = 115-120 ° C, [a6 + 136.5 ° (c 0.32 water); Mass spectrum: (M) m / e477, (M + 1) + m / e 478; Monosaccaride m / e 157 - purple-risamin A-ion, m / e 160,142-garosamine-ion, m / e 191,173,163, 145-5-Epi-2-deoxystreptamin-ion. Disaccharides 350,322,304, 347,319,301.

NMR: 0 (100 MHz, D2O)

5.08, d, J = 3.8 Hz

H'-l 'and H-l

4.99 d, J = 3 Hz

4.39, broad singlet

H-5

3.93 d, J = 12.5 Hz

H-5 "eq

3.77, dd, J = 11, J = ~ 3.6

HzH-2 "

3.30, d, J = 12.5 Hz

H-5 "ax

2.66, d, J = 10.5 Hz

H-3 "

2.53, singlet

N-CH3 (3 ")

2.33, singlet

N-CH3 (6 ')

2.05, m

H-2 eq

1.23, s

C-CHs (4 ")

1.04, d, J = 7 Hz

CH-CH3 (6 ')

30th

35

C. The N and O protecting groups in the intermediates prepared according to Example 5 will be indicated by heating the intermediates with 1 to 2N sodium hydroxide at 100 ° C until thin layer chromatography analyzes of aliquots of the reaction mixture to indicate that the protecting groups have been removed (usually 24 to 48 hours ), removed The product obtained is isolated and purified as described in Example 5B.

D. The N and O protecting groups can be removed from the intermediates prepared according to Example 5A in the following manner. The product according to Example 5A is dissolved in a mixture of 10 ml of tetrahydrofuran and 50 ml of liquid ammonia. 2 g of sodium are slowly added to the stirred mixture and the mixture is continuously stirred for 2 hours. The ammonia is allowed to evaporate by warming to room temperature overnight. The resulting residue is dissolved in 10 ml of 5% sodium hydroxide and heated at 100 ° C for 4 h. The solution is cooled and passed through IRC-50 (H + 1> resin. The resin is washed well with water and the product eluted with 100 ml of 1N ammonium hydroxide. The ammonium hydroxide eluate is concentrated to a residue and this residue becomes as described in Example 5B, 5-epigentamicin Ci is obtained.

Example 6: 50

Other 5-epi-4,6-di-0- (aminoglycosyl) -2-deoxystreptamines

A. 1. In a manner similar to that described in Examples 5A and 5B, each of the following aminoglycoside derivatives is treated with dimethylformamide at reflux temperature,

then each of the O-protected-N-protected-55 intermediates hydrogenated in acetic acid in the presence of palladium on charcoal and finally each of the resulting 2 "-0-benzoyl-3", 4 "-N, 0-carbonyl-5-epiaminoglycosides treated with sodium hydroxide at 100 ° C:

60

l, 3,2 ', 6'-tetra-N-benzyloxycarbonyl-5-0-methanesulfonyl-2 "-0-

benzoyl-3 ", 4" -N.O-carbonylgentamicin C] a,

l, 3,2 ', 6'-tetra-N-benzyloxycarbonyl-5-0-methanesulfonyl-2 "-0-

benzoyl-3 ", 4" -N.O-carbonylgentamicinC2,

1,3,2 ', 6' -Tetra-N-benzyloxycarbonyl-5-0-methanesulfonyl-2 "-O- 65

benzoyl-3 ", 4" -N, 0-carbonylgentamicinC2a,

l, 3,2 ', 6'-tetra-N-benzyloxycarbonyl-5-0-methanesulfonyl-2 "-

0-benzoyl-3 ", 4" -N, 0-carbonylgentamicinC2b.

Each of the products obtained is isolated and purified as described in Example 5B, giving: 5-epigenamicin Qa, 5-epigentamicin C2, 5-epigentamicin C2a or 5-epigentamicin C2b-

2. After treatment of each of the starting materials according to Example 6A with dimethylformamide at reflux temperature, the protective groups in each of the intermediates thus formed can be removed by treatment with sodium hydroxide according to the method of Example 5C or by reduction with sodium in ammonia and subsequent treatment with sodium hydroxide according to Example 5D .

B.l. In a manner similar to that described in Examples 5A and 5D, each of the following aminoglycoside derivatives is treated with dimethylformamide at reflux temperature, then the resulting N-protected-O-protected intermediate with sodium in liquid ammonia and then with sodium hydroxide at 100 ° C treated:

1,3,2 ', 6' -T etra-N-benzyloxycarbonyl-5-0-methanesulfonyl-2 "-O-

benzoyl-3 ", 4" -N, 0-carbonylsisomicin,

l, 3,2 ', 6'-tetra-N-benzyloxycarbonyl-5-0-methanesulfonyl-2 "-0-

benzoyl-3 ", 4" -N, 0-carbonylverdamicin,

1,3,2 ', 6'-tetra-N-benzyloxycarbonyl-5-0-methanesulfonyl-2 "-0-

benzoyl-3 ", 4" -N, 0-carbonyl antibiotic G-52,

l, 3,2 ', 6'-tetra-N-benzyloxycarbonyl-5-0-methanesulfonyl-2 "-0-

benzoyl-3 ", 4" -N, 0-carbonyl antibiotic 66-40D,

l, 3,2 ', 6', 3 "-penta-N-benzyloxycarbonyl-5-0-methanesulfonyl-

2 ", 4" di-0-benzoyl antibiotic 6640B.

Each of the products obtained is isolated, purified and purified as described in Example 5D, giving: 5-episisomicin mp = 135-138 ° C (dec.)

5-epiverdamicin mp = 110-113 ° C [a] g6 + 159.7 (H2O) 5-epi antibiotic G-52,

5-epi antibiotic 6640D or

5-epi antibiotic 6640B.

2. After treatment with dimethylformamide at the reflux temperature, the protective groups in each intermediate product thus formed can be removed by treatment with sodium hydroxide according to Example 5C, the corresponding 5-epiaminoglycoside being obtained.

C. 1. In a manner similar to that described in Examples 5A and 5B, each of the following aminoglycoside derivatives is treated with dimethylformamide at the reflux temperature, then each of the intermediates obtained is hydrogenated in acetic acid over palladium on charcoal and then the product obtained is treated with sodium hydroxide:

l, 3,2 ', 3 "-tetra-N-benzyloxycarbonyl-5-0-methanesulfonyl-3', 2", 4 "-tri-0-benzoyl4 ', 6'-0-benzylidene gentamicin A, 1,3-di -N-benzyloxycarbonyl-5-0-methanesulfonyl-2 ', 3' -O-cycIo-hexylidene-6 ', 4'; 3 ", 4" -di-N, 0-carbonyl-2 "-0-benzoylgentamicin B ,

1,3-di-N-benzyloxycarbonyl-5-0-methanesulfonyl-2 ', 3' -O-cyclo-hexylidene-6 ', 4'; 3 ", 4" -di-N, 0-carbonyl-2 "-0-benzoylgentamicin Bi,

l, 3,2'-tri-N-benzyloxycarbonyl-5-0-methanesulfonyl4 ', 6'-0-cyclohexylidene-2 "-O-benzoyl-3", 4 "-N, 0-carbonylgentamicin X2,

1,3,2 '-Tri-N-benzyloxycarbonyl-5-0-methanesulfonyl4', 6 '-O-cyclohexylidene-2 "-0-benzoyl-3", 4 "-N, 0-carbonyI antibiotic G418,

l, 3,2 ', 6'-tetra-N-benzyloxycarbonyl-5-0-methanesulfonyl-3', 4'-0-benzylidene-2 "-0-benzoyl-3", 4 "-N, 0-carbonyl Antibiotic JI-20A,

1,3,2 ', 6' tetr aN-benzyloxy carbonyl-5-O-methanesulfonyl-3 ', 4' -0-benzylidene-2 "-0-benzoyl-3", 4 "-N, 0-carbonyl Antibiotic JI-20B,

13

618986

1,3,2 ', 6', 3 "-penta-N-benzyloxycarbonyl-5-O-methanesulfony I-3 ', 4'; 4", 6 "-di-0-benzylidene-2" -0-benzoylkanamycin B ,

1,3,2 ', 6', 3 "-penta-N-benzyloxycarbonyl-5-0-methanesulfonyl-4 ', 2" -di-O-benzoyl-4 ", 6" -O-benzylidentobramycin, 1,3 , 6 ', 3 "-Tetra-N-benzyloxycarbonyl-5-0-methanesulfonyl-2', 3 '; 4", 6 "-di-0-benzylidene-4', 2" -di-0-benzoylkanamycin A in Mixture with l, 3,6 ', 3 "tetra-N-benzyloxycarbonyl-5-0-methanesulfonyl-3', 4 '; 4", 6 "-di-0-benzylidene-2', 2" - di-0-benzoylka-namycin A,

l, 3,2 ', 6', 3 "-penta-N-benzyloxycarbonyl-5-0-methanesulfonyl-2" -0-benzoyl4 ", 6" -0-benzylidene-3 ', 4'-di-deoxykanamycin B .

2. Each of the O-ylidene-5-epiamino-glycosides obtained according to Example 6 Cl is dissolved in 50% aqueous acetic acid and the solution is heated on a steam bath for 1 hour. The reaction mixture is evaporated in vacuo to a residue and contains the corresponding 5-epiaminoglycoside. Each of these compounds is purified by chromatographic methods as described in Example 5B, whereby:

5-epigentamicin A,

5-epigentamicin B,

5-epigentamicin Bi,

5-epigentamicin X2,

5-epi antibiotic G418,

5-epi antibiotic JI-20A,

5-epi antibiotic JI-20B,

NMR: (D2O ext. TMS): 5.17 (d. J = 4 Hz, 1 "-M); 5.13 (d. J = 3 Hz, 1'-H); 2.62 (N-Me ); 1.20 (d.J-Hz, Ce -'-CHa); 1.20 5 (Ct, -CH3).

CMR: (D2O, diox, réf.): 102 (Ci '): 96.4 (Ci "); 85.8 and 80.5 ppm

(Ct and Ce).

5-epikanamycin B,

5-epitobramycin,

5-epikanamycin A or

5-epi-3 ', 4'-dideoxykanamycin B.

Example 7:

5-episisomicin and 1-N-alkyl-5-episisomicin A. 5-episisomicin

1. To 10 ml of dimethylformamide 1.2 g of 1,3,2 ', 6'-tetra-N-benzyloxycarbonyl-5-0-methanesulfonyl-2' '-O-benzoyl-

3 ", 4" -N, 0-carbonylsisomicin and 1.0 g of tetra-n-butylammonium acetate are added. The mixture is heated at 120 ° C. for 16 h, evaporated to a residue and the residue is extracted with chloroform. The chloroform solution is extracted with water washed, dried over sodium sulfate and then in vacuo to a residue containing l, 3,2 ', 6'-tetra-n-benzyloxycarbo-nyl-5-epi-0-acetyl-2 "-0-benzoyI-3" , 4 '' -N, 0-carbonylsisomicin, evaporated.

2. The one according to Example 7A1. the residue obtained is dissolved in 10 ml of dimethyl sulfoxide. A solution of 2 g of potassium hydroxide in 4 ml of water is added and the mixture is heated at 100 ° C. for 24 hours. The reaction mixture is cooled, 80 ml of Amberlite IRC-50 (H +) are added, the mixture is stirred for 1 hour, the resin is separated, washed with water and then eluted with 10N ammonium hydroxide. The combined ammonium hydroxide derivatives are concentrated in vacuo and the residue obtained is chromatographed over 25 g of silica gel and eluted with the lower layer of a 2: 1: 1 chloroform / methanol / 10N ammonium hydroxide solvent system. The similar eluates containing 5-episisomicin determined by thin layer chromatography are collected and the combined eluates are evaporated to a residue of 5-episisomicin. Mp = 135-13 ° C (dec.); [a6 + 187.3 (D2O); Mass spectrum (M) + m / e 447, (M + 1) + m / e 448. Monosaccharides m / e 160,127.2-deoxy-5-epistreptamine m / e 191,173,163,145. Disaccharides m / e 317,289,271, m / e 350,322,304.

PMR (a) D20:

5.14

d, J = 2.5 Hz 1'-H

5.07

d, J = 4.0 Hz

1 "-H

4.89

broad singlet

4'-H

4.37

broad singlet

5-H

3.94

d, J = 12.5 Hz

5 "e-H

3.77

q-

2 "-H

3.39

d, J = 12.0 Hz

5 '' a-H

3.21 (2H)

broad singlet

6'-H

2.65

d, J = 11 Hz

3 "-H

2.52

Singlet

3 "-N-CH3

1.23

Singlet

4 "-C-CH3

CMR (D20):

PPM: ct 150.3.102.6.97.1 (2C), 85.8.80.9.73.3.70.3.69.7.68.5.64.0, 48.1.47, 2,47,1,43,2,37,7,36,4,25,6,22,4.

B. 1-N-Ethyl-5-episisomicin

1. The required intermediate, d. H. 1-N-ethyl-l, 3,2 ', 6'-tetra-N-benzyloxycarbonyl-5-0-methanesulfonyl-2 "-0-benzoyl-3", 4 "-NO-carbonylsisomicin is obtained by reaction of 1-N-Ethylsisomicin prepared according to the method of Examples 1 to 4.

In a similar manner to that described in Example 7 A, 1 -N-ethyl-1,3,2 ", 6 '-tetra-N-benzyloxycarbonyl-5-O-methansul-fonyl-2" -O-benzoyl-3 is obtained '', 4 '' -NO-carbonylsisomicin in dimethylformamide treated in the presence of tetra-n-butylammonium acetate at 120 ° C for 16 h. The resulting 5-epi-O-acetate derivative is separated off as described in Example 7A1, then this derivative is treated with aqueous potassium hydroxide in accordance with Example 7A2 and then purified using the chromatographic methods described, and 1-N-ethyl-5-episisomicin is obtained. Mp = 118-122 ° C (dec.); Mass spectrum (M) + m / e 475, (M + 1) + m / e 476. Monosaccharides m / e 160.127. 1-N-ethyl-2-deoxy-5-epistreptamine m / e 219,201,191, 173. Disacccaride m / e 345,317,299, m / e 378,350,322.

PMR (ct) D20:

5.14

d, J = 2.5 Hz 1'-H

5.0

d, J = 4.1 Hz

1 "-H

4.9

broad singlet

4'-H

4.38

broad singlet

5-H

3.93

d, J = 12.5 Hz

5 "e-H

3.78

q-

2 "-H

3.38

d, J = 12.5 Hz

5 "a-H

3.21 (1H)

broad singlet

6'-H

2.65

d, J = 11.0 Hz

3 "-H

2.52

Singlet

3 "-N-CH3

1.22

Singlet

4 "-C-CH3

1.07

Triplet

I-N-CH2-CH3

CMR (D20):

PPM: 149.8, 102.9,

97.4.97.0.83.9.80.5.73.2.70.1.69.6.68.5.63.9.

54,5,47,1,47,0,43,1,40,8,37,5,33,0,25,6,22,4,14,6.

C. Other 1-N-alkyl-5-epiaminoglycosil derivatives

1. In a similar manner to that described in Example 7A, the corresponding 5-epi-O-acetyl derivative gives: 1-N-propyl-5-episisomicin,

1 -N- (n-butyl-5-episisomicin,

lN- (CT-aminobutyl) -5-episisomicin, lN- (Y-aminopropyl) -5-episisomicin, lN- (ß-methylpropyl) -5-episisomicin, lN- {n-pentyl) -5-episisomicin, 1- N <y-methylbutyl) -5-episisomicin, lN- (ß-methylbutyl> 5-episisomicin, lN- (ß, ß-dimethylpropyl) -5-episisomicin, lN <ß-ethylbutyl> 5-episisomicin,

5

10th

15

20th

25th

30th

35

40

45

50

55

60

65

618986

14

l-N- (n-octyl) -5-episisomicin, the treatment of mastitis in cattle and Salmonella,

l-N- (β-propenyl) -5-episisomicin, which can be found in pets, e.g. B. the dog and cat, diarrhea l-N- (ß-ethyl-ß-hexenyl) -5-episisomicin cause.

l-N-Benzyl-5-episisomicin, The improved spectrum of the compounds of this invention

l-N-phenethyl-5-episisomicin, 5 dung consists in the increased effectiveness against many organisms

1-N-CyclohexyhnethyI-5-episisomicin, men, which are resistant to the parent compound. -

l-N- (5-hydroxybutyl) -5-episisomicin, for example the compounds are more active against many organisms.

l-N- (a) -hydroxyoctyl) -5-episisomicin, which the parent antibiotics by acetylation of the l-N- (ß-aminoethyl) -5-episisomicin, 3-amino group and / or by adenylylation of the 2 "-hydro-

Inactivate 1-N-ethyl-5-epigentamicin Qa, 10 xyl group. Some also show activity against 1-N-ethyl-5-epigentamicin Ci, protozoa and amoebas and anthelmintic properties.

1-N-ethyl-5-epigentamicin C2, the 1-N-alkyl derivatives, in particular the 1-N-ethyl-5-epi-4-0-

1-N-Ethyl-5-epigentamicin C2a, aminoglycosyl-6-0-garosaminyl-2-deoxystreptamine, show 1-N-ethyl-5-epigentamicin C2b, also improved activity against Pseudomonas in use

l-N-ethyl-5-epi antibiotic G-52, 15 to their 1-N-unsubstituted precursors with the normal

1 -N-ethyl-5-epiverdamicin or paint configuration on the C-5 atom.

1-N-ethyl-5-epi antibiotic 66-40D. In general, the dose to be administered depends on deri

2. An equivalent amount of the 1-N-ethyl derivative of the 4,6-di-0- (aminoglycosyl) -2-deoxystreptamine of 5-0-methanesulfonyl-2 "-0-benzoyl-0-ylidene-N-benzyloxycarbo The age and weight of the animal species to be treated, the type of nyl intermediates is administered according to Example 7A1 with di-2o administration and the type and severity of the methylformamide to be prevented in the presence of tetra-n-butylammonium or bacterial infection to be attenuated -Acetate treated, then the resulting 5-epi-O-acetylnene is the N-protected-O-protected-intermediate to combat a bacterial infection with an aqueous turning dose of the derivatives of 4,6-di-0- ( aminoglycosyl) -2-potassium hydroxide according to Example 7A2, treated and then deoxystreptamine similar to the required dose of the 0-ylidene-5-epi-4,6-di-0- (aminögly- 25-speaking 4,6-di -0- (aminoglycosyl) -2-deoxystreptamine. Cosyl) -2-deoxystreptamine with 50% aqueous acetic acid acid The derivatives of 4,6-di-0- (aminoglycosyl) -2-deoxystrept-treated according to Example 6C, giving: amines and the pharmaceutically acceptable acid addition

l-N-ethyl-5-epi-3 ', 4'-dideoxykanamycin B, salts can be administered orally. You can also

IN-ethyl-5-epigentamicin B, borrowed in the form of ointments, hydrophilic or hydrophobic, in the form of IN-ethyl-5-epigentamicin Bi, 30 lotions which are aqueous or non-aqueous or as IN-ethyl-5-epi antibiotic JI-20A, emulsion may be present, or applied in the form of creams

1-N-ethyl-5-epi antibiotic JI-20B. Pharmaceutical carriers useful for the preparation of such 1-N-ethyl-5-epikanamycin B formulations are e.g. Water, oils,

1-N-ethyl-5-epitobramycin, fats, polyesters, polyols and the like.

1-N-ethyl-5-epi antibiotic 66-40B, 35 For oral administration, the compounds can be

1-N-ethyl-5-epigentamicin X2, this invention for tablets, capsules, elixirs or the like

1-N-ethyl-5-epi-antibiotic G-418, can be processed, or they can also be mixed with animal feed 1-N-ethyl-5-epikanamycin A or. These are the dosage forms in which 1-N-ethyl-5-epigentamicin A. the bactericides are most effective for the treatment of bacterial

The compounds prepared in accordance with this invention and 40 infections of the gastro-intestinal tract which are infectious, whose non-toxic pharmaceutically acceptable acid addenda cause diarrhea.

tion salts are antibacterial agents with a broad spectrum. In general, preparations to be used externally — which advantageously have activity against many organisms of about 0.1 to about 3.0 g of active ingredient per 100 g of the men, compared to their 5-hydroxy precursors Ointment, cream or lotion. Are resistant to external use. The compounds can therefore be used alone or in 45 preparations, usually gently about 2 to about 5 times in combination with other bactericides, applied to wounds day.

to prevent the growth of bacteria in different media or to reduce their number. You can e.g. The bactericides of this invention can be in liquid form, disinfection of laboratory glassware, dental and e.g. B. as solutions, suspensions and the like, can be used in ear medicine facilities which are used with 50 and ophthalmology and can also be infected with Staphylococcus aureus or other bacteria, administered parenterally by intramuscular injection which is inhibited by the aminoglycosides of this invention. The injectable Solution or suspension will usually be in. The effectiveness of the compounds against gramnega amounts of from about 1 mg to about 10 mg of bactericide per kg of body-weight bacteria makes them effective in combating infections, per weight per day, divided into about 2 to about 4 doses, softened by gram-negative organisms, e.g. B. Species of Pro-55 ranges. The exact dose depends on the stage and the teus and pseudomonas that are evoked. The severity of the infection, the sensitivity of the infected compounds, e.g. 5-Epigentamicin Ci and 5-Epigentamicin organism against the bactericide and the individual Cia, are used in veterinary medicine, especially with characteristics of the animal species to be treated.

Claims (10)

618 986 PATENT CLAIMS 1. Process for the preparation of derivatives of 4,6-di-O- (aminoglycosyl) -2-deoxystreptamine Gentamicin A, Gentamicin B, Gentamicin Bi, Gentamicin Ci, Gentamicin Cla, Gentamicin C2, Gentamicin C2a, Gentamicin C2b, Gentamicin X2 , Tobra-mycin, Verdamicin, Kanamycin A, Kanamycin B, 3 ', 4'-Dide-oxykanamycin B, Antibiotic G-52, Antibiotic 6640B, Antibiotic 6640D, Antibiotic G-418, Antibiotic JI-20A, Antibiotic JI-20B and Sisomicin, in which the 2-deoxy-streptamine group is replaced by a 1,3-diaminocyclitol of the formula nh, nhr x 0 is replaced and in the formula R is hydrogen or the group -CH2Y, where Y is hydrogen, alkyl, alkenyl, cycloalkyl, cycloalkylalkyl, hydroxyalkyl, aminoalkyl, N-alkylaminoalkyl, aminohydroxyalkyl, N-alkylaminohydroxyalkyl, phenyl, benzyl or tolyl and the aliphatic radicals have up to seven carbon atoms and, if substituted by amino and hydroxy, have these substituents on different carbon atoms, and X is hydroxy, and their pharmaceutically acceptable acid addition salts, characterized in that a derivative of one of the above 4, 6-di-0- (aminoglycosyl) -2-deoxystreptamine, in which the 2-deoxystreptamine group is replaced by a 1,3-diaminocyclitol of the formula nh, nhr 0 0 and is in the formula R as defined above and X 'represents unsubstituted or substituted hydrocarbon sulfonyloxy, and the hydroxy and amino groups in the 4,6-di-0- (aminoglycosyl> 2-deoxystreptamine derivative are protected by groups, which can be removed by reductive cleavage or basic or weakly acidic hydrolysis, reacted with dimethylformamide in a temperature range from 80 ° C. to 155 ° C., the protective groups removed and the derivative obtained isolated as such or as a pharmaceutically acceptable acid addition salt. 2. The method according to claim 1, characterized in that the reaction with dimethylformamide is carried out in the presence of a tetraalkylammonium alkanoate. 3. The method according to claim 2, characterized in that tetra-n-butylammonium acetate is used as tetraalkylammonium alkanoate. 4. The method according to any one of claims 1 to 3, characterized in that X 'is hydrocarbon sulfonyloxy having up to eight carbon atoms or a halogen derivative thereof or is nitrobenzenesulfonyloxy. 5. The method according to any one of claims 1 to 4, characterized in that X 'represents methanesulfonyloxy. 6. The method according to any one of claims 1 to 5, characterized in that the removal of the protective groups with aqueous base or, if the protective groups can be removed by reductive cleavage, with hydrogen in the presence of a catalyst or with an alkali metal in liquid ammonia and subsequent treatment with aqueous base takes place and, if acetals or ketals are present, these are then removed with aqueous acid. 7. The method according to any one of claims 1 to 6, characterized in that a derivative of sisomicin is used and R in the 1,3-diaminocyclitol of formula II is hydrogen. 8. The method according to any one of claims 1 to 6, characterized in that a derivative of sisomicin is used and R in the 1,3-diaminocyclitol of the formula II is the group -CH2Y and Y is as defined in claim 1. 9. The method according to claim 8, characterized in that the group -CH2Y represents ethyl. 10. Use of the compounds of the formula I, wherein R is hydrogen, prepared by the process of claim 1 for the preparation of compounds of the formula I, in which R is the group -CH2Y, where Y is the in claim 1 has given meaning, characterized in that the compound with an aldehyde of the formula Y-CHO, wherein Y has the meaning given in claim 1, in the presence of a hydrogen donor reducing agent and, if appropriate, with intermediate protection of amino groups in the compound of Formula I with the exception of the amino group in position 1 and, if appropriate with intermediate protection of any amino or hydroxy group present in the aldehyde of the formula Y-CHO, and isolating the compound obtained as such or as a pharmaceutically acceptable acid addition salt.