DE2135191C3 - 3 ', 4'-Dideoxykanamycin B, process for its preparation and medicament containing it - Google Patents

Description

HO

OH

2. Process for the preparation of 3'.4'-dideoxykanamycin B according to claim 1, characterized in that the amino groups of kanamycin B are reacted in a manner known per se with ethyl chloroformate or salicylaldehyde, then the 4 "and 6" hydroxyl groups with acetone or cyclohexanone dimethyl ketal, then the 2 "hydroxyl group is benzoylated in the usual way, from the 4", 6 "-Di-O-isopropylidene or 3 ', 4';4", 6 "-Di- O-cyclohexylidene derivative splits off acetone or cyclohexanone by heating with aqueous acetic acid, converts the 3'- and 4'-position hydroxyl groups in the usual way with methanesulfonic acid chloride into 3 \ 4'-disulfonic acid ester groups, these are converted into the 3 \ 4 'by reaction with sodium iodide and zinc -Didehydroverbindungen transferred, this is catalytically hydrogenated in a manner known per se with hydrogen in the presence of PtO ₂ and finally the remaining protective groups in a manner known per se with dilute acetic acid and then B arium hydroxide removed.

3. Medicaments, characterized by a content of a compound according to claim 1 and customary 1 excipients and / or diluents and / or auxiliaries.

NH,

HO

(Kanamycin: R- OH)
(Kanamycin B: R = NH ₂ )

On the basis of these findings, an attempt was first made to block the 3 'hydroxyl group and to synthesize the 3'-0-methylkanamycin. However, this derivative did not have any antibacterial properties Activity. Then, 3'-deoxykanamycin was synthesized and it was found that this derivative is active against the resistant bacteria described above. Hence became assumed that the replacement of the 3 'permanent hy-

Hydroxyl group inhibits the phosphorylation of kanamycin by resistant bacteria, thereby giving this antibiotic the ability to attack the kanamycin-resistant bacteria.
The highly active 3 ', 4'-dideoxykanamy-

45 cin B synthesized of the following formula:
CH ₂ NH

55

The invention relates to 3 ', 4'-Dideoxykanamy-B, which has antibacterial effects and in particular against infections of kanamycin-1 organisms is effective, a method for his production and medicinal products containing it. It rests on investigations into the mechanism τ have elucidated drug inactivation by bacteria resistant to bacteria from patients and e due to the mechanism to new antibiotic, effective against the resistant bacteria derivatives led.

HO

OH

The process for the preparation of 3 ', 4'-dideoxy-

Le.

Kanamycin B is characterized in that the amino groups of kanamycin B are reacted in a known manner with ethyl chloroformate or salicylaldehyde, then the 4 "and 6" hydroxyl groups are reacted with acetone or cyclohexanone dimethyl ketal, then the 2 "hydroxyl group in Usually benzoylated, acetone or cyclohexanone is split off from the 4 ", 6" -Di-O-isopropy-Hden- or 3 ', 4'; 4 ", 6" -Di-O-cyclohexylidene derivative obtained by heating with aqueous acetic acid , converts the 3 'and 4' hydroxyl groups into 3 ', 4'-disulfonic acid ester groups in the usual way with methanesulfonic acid chloride, converts these into the 3 \ 4'-didehydro compound by reaction with sodium iodide and zinc, this in a manner known per se with hydrogen catalytically hydrogenated in the presence of PtO ₂ and finally the remaining protective groups are removed in a manner known per se with dilute acetic acid and then barium hydroxide.

The synthetic route for the new 3 ', 4'-dideoxykanamycin B is briefly as follows:

Kanamycin B (free base) is used as the starting material. It is treated with the usual means, around the five free amino groups, the 4 ", 6" standing hydroxyl groups and all or part of the 5- and 2 "and the 3'- and 4'-hydroxyl groups of the starting kanamycin B to protection. It. can e.g. B. the following Kanamycin B derivatives are produced:

CH ₂ -NHCOOC ₂ H ₅ NHCOOC ₂ Hs

³¹ NHCOOC ₂ Hs

OH '

OCO

HO

OC

The compound of type I or Γ is then treated with aqueous acetic acid, the 3 'and 4' positions Hydroxyl groups are released. These products are then generalized into derivatives of the following Formula II transferred:

CH ₂ NHCOOC ₂ H ₅ (or N = CH - <^ J />)

NHCOOC ₂ H ₅ (OdCr N = CH- / ~~ *

^ NHCOOC ₂ H ₅ ί eder N = CH ^ f ^N

NHCOOC ₂ Hs
(or N =

H ₁ C

O-CH

(H)

or N = CH

wherein the 3 'and 4' hydroxyl groups have been sulfonylated, ie W means a —SO ₂ CH, group.

If this disulfonylated derivative (II)

(W = SO ₂ CH ₃ )

treated with sodium iodide and zinc dust in a suitable solvent, this is obtained 3,4'-unsaturated derivative, which after reduction with hydrogen in a manner known per se in the presence of platinum oxide as a corresponding hydrogenation product, namely as one protected by amino and hydroxyl groups 3 ', 4' - dideoxykanamycin B is present. When using this derivative according to conventional measurement methods the protective groups are split off, namely with dilute acetic acid and then Barium hydroxide, 3 ', 4'-dideoxykanamines are obtained in B.

The synthetically produced 3 ', 4'-dideoxykanamycin B inhibits the growth of various kinds of bacterial strains in the same manner as kanamycin B, as shown in the table. It also inhibits the growth of kanamycin B-resistant staphylococci, resistant Escherichia coli and resistant Pseudomonas aeruginosa. 3 ', 4'-Dideoxykanamycin B shows a therapeutic effect against infections caused by staphylococci. Klebsieila pneumoniae. Salmonella typhosa and Pseudomonas aeruginosa in mice. 3'.4'-Dideoxykanamycin B has a low toxicity (LD ₅₀ 180 mg / kg mouse, i. Ν.) And shows a high blood level after the injection. It follows that the product of the present invention is a valuable compound in the treatment of various types of infections, including infections caused by gram-negative and gram-positive resistant bacteria. From the above table it can be seen that the product prepared according to the invention shows a strong effect in the inhibition of pathogenic microorganisms.

table

Antibacterial spectra of 3 ', 4'-dideoxykanamycin B and kanamycin B

Test organism ¹ ) ² ) Minimum inhibitory concentration (mcg / ml) 0.78 B Kanamycin B 6 "-deoxy-
kanamyein 3,4-dideoxykanamycin
Sample 1 sample 2 1.56 0J8 1.56 Staphylococcus aureus FDA 209 P 1.25 1.56 1.56 1.56 Escherichia coli NIHJ 2.5 1.56 0.78 1.56 K-12CS-2 2.5 > 50 > 100 K-12 ML 1629 ⁺ ■ 2.5

7 - ⁸

continuation

, Minimum inhibition / cnlration (mcg ml)

csl-Orcanisnuis ¹ η, _η ,., _η

V4-Didcsoxykanamycin B Kanamycin B 6 -deoxy-

'' kanamycin

Sample I Sample 2

K-12 ML 163O ^{+ 2} ' ⁵

K-12 ML 1410 seudomonas aeruginosa A 3 No. 11 ^{+ +} No. 12 ^{+ +} No. 39 ^{+ +}

45 ^{++ U25}

No. 67 ^{+ +}

• rotus rettgeri GN311 ^{+ +} GN466 ^{++ 2} ' ⁵

3.12 > 50 > 100 1.56 0.78 1.56 3.12 50 100 3.12 > 50 > 100 6.25 50 100 6.25 > 50 > 100 0.78 25th 100 6.25 > 50 > 100 12.5 3.12 3.12 3.12 1.56 3.12

') Culture medium agar, 37 ° C, 18 hours. Β, -imnfen with the resistance factor (R-factor) in Escherichia coli K-12

') The strains marked with * were byc »*" »« · * « » ™ ^P g _η ™' _is Ter1e. Compared to many known Anbb.ot.ka res.stente

obtained in vitro; the ^{strains marked with +} * are from patients

Tribes; the rest are natural tribes.

Penta - N - ethoxycarbonyl - 4 ", 6" - O - isopropylidene-

The following examples are preferred _{in the} same way as

Manufacturing process of 3 ', 4'-dideoxykanamycin B ³ ° * £ ™ £ _{eschritten m} the penta-N-ethoxycarbonyl described in detail. y ^,. ^, ₆ ». ^ j - O- isopfopylidenkanamycin B vice

to be changed.

Example! ₂ "-O-Benzoyl-penta-N-ethoxycarbonyl-

. r, 35 ^^ '^ "^". di-O-isopropylidene-canamycin B.

a, Penta-N-ethoxycarbonylkanamyan B ^^ _{Example lb)} produced Kanamy-

To a stirred mixture of 1.0 g of Kanamy · '“T? _erivats were dissolved in 48 ml of pyridine, and a B (free "base) and 0.9 g of anhydrous sodium ^OT" JRR _T .. the _first given 2 g of benzoyl chloride, carbonate in aqueous acetone (1: 1,20 ml) were ^ _a ^ ^ ^g _olution at room temperature for 1 hour 1.05 ml of ethyl chloroformate was added, and ₄ o Man "" ^^^ _was evaporated and it was for 5 hours at room temperature, overall length, a.; , _ch i _oro form solved. This solution was stirred. The precipitate formed was ^Κ ^! °° ™ _{& 5Λε η} and geabfiltriert over sodium sulfate, washed with water and dried. with water 8 it _{was then brought close to} rnd

1.46 g of a solid substance were obtained. The melting ξ ™ ^ _{"A and} after addition of n-hexane punktlagbei305 ° C (Zerset _Z ung), [a] l: 88 + ^o (c = 0.5; 20m ^^ as _a crystalline substance with dimethylformamide)? : «JSf £ Ζ * £ * on 205209 ° C [«] Γ: + 114 °

Dimethylformamide): «S. Melting point of 205-209 ° C, [«] ϊ: + 114 °

b) Penta-N-ethoxycarbonyl-3 ', 4'; 4 ", 6" -di-O-iso- _{(c =} ι _; dimethylformamide).

propylidenkanamycin B "j -'- O-benzoyl-penta-N-athoxycarbonyl-13.7 g of the penta-5 ° ^^. o-isopropylidenkanamycin B N-ethoxycarbonylkanamycins B prepared according to Example la) were in 70 ml diet -. -r , Example 1 c) dissolved Kanamymethylformamid, and the solution was after. ^ fj ^ vats were in aqueous acetic acid addition of 18.8 g of 2,2-dimethoxypropane and 0.35 g of Jf: ¹ ™ « _lös , ^ d the solution was 30 Mmuten anhydrous p-toluenesulfonic acid for 1 hour on γ - ^^ λΖη erVärmt the solution thus prepared 65 ⁰ C heats the solution thus prepared was 55 tong ■ ^a ™ _"ft ^ _the resulting deaatonierte to approximately 50 ml concentrated and after Addition ^ J ^ 'Sg) ¹ J _n 15 ml of dimethylformamide dissolved from 30 g of 24-dimethoxypropane again 1 hour J ^ J ¹¹ "^^ *' _{on 0} , 65 g of 2,2-dimethoxypropane and heated ^{to 65 2 C for a long time Daim} were 6 ml Tnalhyl- After ^z ^^? _a 'xoTuoisuifonsäure allowed to the amine was added and the solution was poured into a stirred »™ b ι hour at room temperature mixing" 500 ml of benzene and 500 ml Wass ^ 6o Lo ^ gJ hour ^ j ^ _fa ^^ ^ poured It was a solid precipitate *? a * nv Precipitates were filtered off, (solid A). After the solid A has been filtered off, the ^ε ° ™ characteristic _Λεη and treated with benzene, the benzene layer is dull. The resulting "JL ^W ^ _t ^g _2j oig a benzene-insoluble solid precipitates were filtered, washed with benzene and water ^ ⁶ St melting at 28S-287 ° C ser washed and dried to give f 3 5g 65 substance ™ * ^m _{= 1 ;} Dimethylformamide); NMR of a solid with a melting point of 236 WiL _n L ¹¹ L _{Dimethylsulfoxid} U ₆ ): τ8.72 and 8.60 to 23TC, Ca] ?: + 87 ° (c = 1; Dimethylformamide ^ spectrum ^ J ^ ^ _{] eXt is} isopropylidene-Pro- The resulting solid A (8.1g) was found to be (each 3 "cn

tones), τ 1.5-2.8 (5-proton-Multiplelt is assigned to benzoyl protons).

e) 2 "-0-Benzoyl-penta-N-ethoxycarbonyl-4", 6 "-O-isopropylidene-3 ', 4'-di-O-mesylkanamycin B.

1.18 g of the kanamycin B derivative prepared according to Example Id) were dissolved in 15 ml of pyridine, and the solution was left to stand for 1.5 hours at room temperature _{after adding 0.66 g of methanesulfonyl chloride (CH 3} SO _{2 Cl).} The solution was evaporated to one third of the original volume and the solution thus prepared was poured into water. The resulting precipitates were filtered off and washed with water. Of a solid was obtained 1.32 g with a melting point of 198 ⁰ C, [a] £: + 107 ° (c = 1.5; dimethylformamide); NMR spectrum (in pyridine d ₅ ): τ 6.60 and 6.45 (each 3 proton singlet can be assigned to mesyl protons).

0 2 "-O-benzoyl-3 \ 4'-dideoxy-3 ', 4'-didehydro-

penta-N-athoxycarbonyW.o ^ O-isopropylidene-

kanamycin B

11 g of sodium iodide and 5 g of zinc dust were added to a solution consisting of 1.0 g of the kanamycin B derivative prepared according to Example 1 e) and 20 ml of dimethylformamide, and the mixture was heated to 95 ^° C. for 1 hour with vigorous stirring heated. The mixture was then poured into a hot mixture of 100 ml of chloroform and 100 ml of water with stirring, and the isolated chloroform layer was washed with water, dried over sodium sulfate. After evaporation, 940 mg of a solid were obtained. The solid was treated with hot ethyl acetate and 310 mg of the insoluble solid was recrystallized from a mixture of methanol-chloroform-ethyl acetate. A substance was obtained with a melting point of 282-284 ° C, [a] S ^? : +36 ^C (c = 0.4; dimethylformamide); NMR spectrum (in pyridine-d ₅ ): τ 4.06 (2 protons in a somewhat broadened singlet, to be assigned to hydrogen in 3- and 4'-position).

Any precipitates were filtered and dried, and 750 mg of a substance with a melting point of 247-248 ° C. were obtained, [«] ?: +89 ' (c = 2.7; dimethylformamide). Found C: 52.41; 5 H: 6.54; N: 7.48%. For C ₄₀ H ₆₁ N ₅ O ₁₀ C: 52.45; H: 6.71; N: 7.65%.

i) 3 ', 4'-Dideoxykanamycin B

150mg of the Kana-ίο produced according to example lh) mycin-B denvate and 2.3 g of barium hydroxydoctanydrate were added to a mixture of 5 ml of water and added 4 ml of p-dioxane. The mixture so prepared was on a boiling point for 8 hours Heated water bath. Then carbon dioxide was

'5 passed, and the resulting precipitates were removed. The supernatant layer was isolated and evaporated. The residue obtained in this way was dissolved in water, and the solution was after H tration on a with "Amberlite IRC-50" (H ⁺ form)

loaded column with 0.1-0.4N ammonium hydroxide chromatographies. The fraction containing 3 ', 4'-dideoxykanamycin B was concentrated. By adding acetone, 47 mg of a crystalline substance were obtained. ["] * _: + 130 ° (c = 0.65; water).

Found: C: 47.71; H: 8.38; N:! 5.31%. Calculated for C ₁₈ H ₃₇ N ₅ O ₈ : C: 47.88; H: 8.26: N: 15.51%.

g) 2 "-O-benzoyl-3 ', 4'-dideoxy-penta-

N-ethoxycarbonyl-4 ", 6" -0-isopropylidene-

kanamycin B

1.15 g of the kanamycin B derivative prepared according to Example IQ. ie des 2 "-O-benzoyl-3 ', 4'-dideoxy - 3', 4 '- didehydro - penta - N - ethoxycarbonyl-4", 6 "-O-isopropylidene-kanamycinsB, which in a mixture of 45 ml p-Dioxane, 25 ml of methanol and ml of water had been dissolved, was hydrogenated with hydrogen at 2 at pressure and in the presence of 0.3 g of platinum oxide, which had previously been activated, at room temperature for 5 hours. The solution thus prepared was filtered and . NMR spectrum (in pyridine-dj) + 87 ° (c = 0.2;; dimethylformamide): evaporated to give 1.10 g of a colorless powder having a melting point of 254-256 ⁰ C, [u] S ' r 7.8-8.5 (4 protons, broad signal, to be assigned to the hydrogen in the 3 ', 4 -position).

h) 2 "-O-benzoyl-3 ', 4'-dideoxy-penta-N-ethoxycarbonyl-kanamycin B.

980 mg of the kanamycin B derivative prepared according to Example 1 g) were kirted in a mixture of ml of acetic acid and 5 ml of water. The solution was heated for 5 minutes at 10O ⁰ C the ent-Example 2 ^a> Penta-N-B salicylidenkanamycin

To a suspension of 400 mg of kanamycin B base in aqueous methanol (1: 8, 20 ml) were added 3 ^ mg of saiicylaldehyde added, and the resulting The solution was poured into water. The precipitates were filtered, dried, and yielded 750 mg penta-N-salicylidene-canamycin B.

b) 3 ', 4'; 4 ", 6" -Di-O-cyclohexylidene-penta-N-salicylidene-kanamycin ^{BN 7 ür m} L ^de ? ^{according to} Example 2a) provided dental salicylidenkanamycins B were dissolved in 12 ml of dry dimethylformamide, and the solution was after the addition of 500 mg of cyclohexanone dimethyl ketal and 30 mg of anhydrous p-toluenesulfonic acid ι hour

teng heated to 5O ⁰ C. Then a small amount of the solvent was removed by evaporation in vacuo. The solution thus prepared was poured into water. The resulting ^ low

710 m ^{were filtered} '^ dries and yielded

c) 2 "-O-Benzoyl-3 ^> , 4 ';4", 6 "-di-O-cyclohexylidene-

penta-N-salicylidene-canamycin B

«™ ⁵⁹ - ^0mg ~ ^oF b ^of Example 2) produced kanamycin-B-Denvats were dissolved in 10 ml of pyridine, and to the solution were added 120 mg of benzoyl chloride *. The mixture was left to stand for 1 hour at low temperature, then the solution was poured into water. The resulting precipitates were filtered off and dried, and a yield of 612 mg was obtained.

d) ² "-O-Benzoy] -4", 6 "-O-cyclohexylidene-penta-

N-ethoxycarbonylkanamycin B

h ^A mixture of 600 mg of the kanamycin B derivative prepared according to Example 2c) and aqueous oleic acid (1: 3, 20 ml) was heated to * ·><- for 30 minutes. The reaction mixture was

evaporated and the residue treated with ether. The ether-insoluble part was dissolved in aqueous acetone (1: 1, 15 ml). After addition of 400 mg of sodium carbonate, 270 mg of ethoxycarbonyl chloride were added to the solution added. It was then stirred for an additional 5 hours at room temperature. The resulting Precipitates were filtered off, washed with water and dried. 515 mg were obtained a solid substance, [«] ?: + 98 ° (c = 1.5: dimethylformamide). This substance could in a manner similar to that described in Examples Ie) to Ii). be converted into 3'.4'-dideoxykanamycin B.

Claims (1)

Patent claims: 1. 3 ', 4'-Dideoxykanamycin B of the formula
H ₂ NH ₂ C NH, It was found that drug-resistant gram-negative bacteria from patients who carry d? n R-factor, resistant staphylococci and Resistant Pseudomonas aeruginosa produce an enzyme that inactivates kanamycin by removing the phosphate residue is transferred from adenosine triphosphate to the 3'-position hydroxyl group of the kanamycin. CH ₂ NH ₂ IO ■ S