DE2135191A1 - - Google Patents

Description

MW Heme, MOB Munich 25,

rrälllgrathstrasse 1? ! »ΓαΙΙ ··« »* !! *» »* · Elsenacher Strasse 17

Posnach 140 Dipl.-1 ng. κ. ti. ua »ir m ***. ***,

P »t-Anw. HenmMtn-TmttepoM DIP I. - PHV S. E d U α Γ α BetZler F »m» _P recher: Ϊ9 8011

Telephone: 51013 ~ 3 »8012

διό «DIpl.-Ing. W. Herrmann-Trentepohl »» »

Tetegrammansdirlfl: fHTCHT «uuixiTc Telegremmanschrift:

Bahr patents Mr. PATENTANWÄLTE Babetzpal Munich

Telex 08229853 Telex 05 245 42 I I bank accounts:

Bayrische Vereinsbank Munich 952 287 Dresdner Bank AG Herne 202 456 Postal check account Dortmund 558 48

L J

Ref .: _M0 2987

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Munich

ZAIDAN HOJIN BISEIBUTSU KAGAKU KENKYU KAI No. 14-25, 3-chome, Kami-Osaki, Shinagawa-ku,

Tokyo / Japan

Process for the preparation of ^ '»V-dideoxykanamycin B

The invention relates to the production or synthesis of ^ ', V-Dideoxykananiycin B, the antibacterial effect and is particularly active against infection of kanamycin-resistant organisms. The present invention is based on studies that have elucidated the mechanism of drugs in activation by resistant bacteria and the active derivatives due to the mechanism of new antibiotic bacteria resistant to the drug Looking for.

It was found that drug and drug resistant gram-negative bacteria caused by the

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R patent-based pactors carry resistant Staphylococci and resistant Pseudomonas aeruginosa, producing enzymes that inactivate kanamycin by transporting phosphate from the ATP to the 3'-hydroxyl group of the kanamycin.
6 '

2 "

y OH
Kanamycin R-OH
Kanamycin BR = NH2

Based on these findings, an attempt was first made ^{to block the 3 f} -hydroxyl group and the 3'-0-methylkanamycin was synthesized. However, this derivative would have no antibacterial activity. 3 ^f -deoxykanamycin was then synthesized and it was found that this derivative is active against the resistant bacteria described above. Therefore, it was believed that substitution of the hydroxyl groups, which are phosphorylated by the resistant bacteria, with oxygen would enable the antibiotic to attack the resistant bacteria. This principle was used with Kanamycin B and the very active one

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^ ', V-dideoxykanainyein B ,. has the following formula, was synthesized.

3 "OH

The synthetic route is as follows: Kanamycin B is used as the starting material, which is converted into the kanamycin B derivative, as shown in the general formulas (I) or (I ¹ ), in which the five amino groups 4 ", 6 ^M Hydroxyl groups and all or part of the 5.3 *, V and 2 "hydroxyl groups are protected.

CH ₂ KHCOR

NHCOR 3 j. NHCOR

(D

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N = CHE '

A.D

(I ¹ )

R = H, alkyl-> aralkyl-, aryl-, alkyl oxy-, aralkyloxy- or Aryloxy groups.

R ^f = H, alkyl, aralkyl or aryl groups.

A.
X = H, H or represented \ _c ^ where A and B

B ^
H, alkyl or aryl groups or

(CH ₂ ) _n
Y 1st

C:

(n = 4.5 or 6).

(where A and B, H, alkyl or aryl

Groups or (CH2) _n C (n = 4.5 or 6).

Z = H, acyl, aralkyl, aryl groups or alkylsulfonyl, aralkylsulfonyl or arylsulfonyl groups.

The derivative (I) or (I ¹ ) is then converted into the derivative shown in the general formula (II) (W = H), in that the j5 ^f and 4'-hydroxyl groups have been freed. This derivative is then converted to derivative (II) represented in the following formula:

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CH ₂ NHCO R (or N = CHR ') 0

NHCOR

(or N = CHR ¹ )

NHCOR (-or N = CHR ¹ )

NHCOR (or N = CHR ¹ ).

(H)

OH ₂ C

OZ

W = H, alkylsulfonyl, aralkylsulfonyl or arysulfonyl groups. .

R, R ^! , Y and Z are the same substituents as shown in the general formula (I) or (I ^f ).

From the general formula II (W = SO ₂ Qi Qi alkyl, aralkyl or aryl groups) that the j3 ^f and 4 ^ hydroxyl groups are sulfonated (sulfonylated). The derivative (II) (W = SO ₂ Q) is then treated with iodine ions and a metal powder which has reducing properties. Such a metal powder is zinc dust, for example. The reaction is carried out in a suitable solvent and gives a derivative which has a 3 '* 4' unsaturated group. The protective groups of the derivative prepared in this way are removed in a conventional manner after hydrogenation of the unsaturated groups and give the end product 3 ¹ , 4 ^I -dideoxykanamycin B.

The synthetically produced 5 ^f , 4'-dideoxykanamycin B inhibits the growth of various kinds of bacterial strains in the same way as the kanamycin B.

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as can be seen from Table 1 and also inhibits the growth of the kanamycin and kanamycin B resistant Staphylocöcei, the resistant Escherichia coli and the

€ 3

resistant Psudomonas aeruginosa. 3 ^f , 4 ^f -Dideoxykanamycin B shows a therapeutic effect against infections * caused by Staphylocöcei, Klebsiella pneumoniaej Salmonella typhdsa and Pseudomonas aeruginosa in mice. 3 ^s * 4'-dideoxykanamycin B has a low toxicity (LDc l8o ₀ mg / kg mouse, iv.) And shows a high blood level after injection. As a result, the derivative of the present invention is a valuable and beneficial chemotherapeutic product for the treatment of various types of infections, including infections produced by gram-negative and gram-positive resistant bacteria. From the above table it can be seen that the derivative prepared according to the invention shows a strong effect in the inhibition of microorganisms, including pathogenic organisms.

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Antibacterial spectra of 3 ', 4 ^t -dideoxykanamycin B and kanamycin B

Minimal inhibiting Sample 2 concentration (mcg / ml) Test organism ^x Dideoxykanamycin B 0.78 Kanamycin B Sample 1 1.56 Staphylococcus 1.25 1.56 0.78 aureus PDA 2o9 P 1.56 Escherichia coli , 2.5 3.12 1.56 NIHJ ₍ '2.5 1.56 0.78 K-12 CS-2 2.5 35o K-12 ML 1629 2.5 3.12 > 5o K-12 ML I630 3.12 0.78 K-12 ML I4lo 6.25 Pseudomonas 2.5 6.25 5o aeruginosa A3 0.78 > 5o No. 11 6.25 5o No. 12 > 5o No. 39 1.25 12.5 25th No. 45 3.12 > 5o No. 67 Proteus rettgeri . · 3.12 GN 3II 2.5 1.56 GN 466

χ Culture medium agar, 37 ° C, 18 hours

^{Preferred types of preparation of 3 1} , 4 * -dideoxykanamycin B are described in detail in the following examples.

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example 1

Synthesis of penta-N-ethoxycarbonkanamycin B.

To a stirred mixture of 1.0 g of Kanamycin B (free base) and 0.9 g of anhydrous sodium carbonate in aqueous Acetone (1: 1.20 ml) was 1.05 ml of ethoxycarbonyl chloride added and stirred for five hours at room temperature. The resulting precipitate was filtered off, washed with water and dried. 1.46 g of a solid substance were obtained. The melting point was (Decomposition.), ΒQ 15 + 88 ° (c 0.5 * dimethylformamide).

Example 2 - -

Synthesis of penta-N-ethoxycarbonyl-J '- ^ ¹ ;

13 * 7 S of the penta-N-Äthoxycarbonylkanamycin B prepared according to Example 1 were dissolved in Jo ml of dimethylformamide and the solution was after the addition of 18.8 g of 2,2-DlmethQXypropan and 0.35 g of anhydrous p-toluenesulfonic acid for one hour 65 ⁰ C heated. The solution prepared in this way was concentrated to approximately 50 ml and, after the addition of 150 g of 2,2-dimethoxypropane, was again heated at 65 ° C. for one hour. Then 6 ml of triethylamine were added and the solution was poured into a stirred mixture of 500 ml of benzene and 500 ml of water. A solid precipitate formed (solid A). After the contaminant A was removed by filtration, the benzene layer was allowed to stand. The resulting precipitates were filtered, washed with benzene and water and dried. 3.5 g of a solid were obtained with a melting point of 256 to 237 ° C. QCj Q ° + 87 ⁰ (c 1, dimethylformamide.

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It was found that the obtained pesticide A (8.1 g) penta-N-ethoxycarbonyl ~ 4 ^tl , 6 "-0-isopropylidene-kanamycin B and the compound could be converted into the penta-N- in the same manner as described above. Ethoxycarbonyl-J ', 4 ¹ J 6 "-di-O-isopropylidene-kanamycin B can be converted.

example

Synthesis of 2 "-0-benzoyl-penta-N-ethoxycarbonyl-3 ^l , 4 ^l ; 4", 6 "-di-0-isopropylidene-kanamycin B {[" of the general formula (I), R = OC ₃ H ₅ , X = Y = CH (CH ^) ₂ , Z at C-2 ". Is COCgH ₅ and Z at C-5 is HJ

5.24 g of the kanamycin B derivative prepared according to Example 2 were dissolved in 48 ml of pyridine, and 2 g of benzoyl chloride were added to the solution. The solution was allowed to stand at room temperature for one hour, the solution was evaporated and the residue was dissolved in chloroform. This solution was washed with water and dried over sodium sulfate. The solution was then concentrated to approximately 20 ml and, after addition of n-hexane, 3 * 35 g of a crystalline substance with a melting point of 2o5-2o9 ° C ζοϋ) ^ ¹ + 114 ° (c!, Dimethylformamide).

example

Synthesis of 2 "-0-benzoyl-penta-N-ethoxycarbonyl ~ 4", 6 "-0-isopropylidene-kanamycin B.

2.82 g of the kanamycin B prepared according to Example 3 Derivative were dissolved in aqueous acetic acid (1: 3 * 40 ml).

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dissolves and the solution was heated J5o minutes at 95 ⁰ C. The solution so prepared was evaporated and the deacetonated product obtained (2.58 g) was dissolved in 15 ml of dimethylformamide. After adding 0.65 of 2,2-dimethoxypropane and 55% anhydrous p-toluenesulfonic acid, the solution was left to stand for one hour at room temperature. The solution was then poured into water and the resulting precipitates were filtered off, washed with water and treated with benzene. This gave 2 g of an oil-benzene-insoluble solid having a melting point of 285-287 ⁰ C. ΰύ I) ^{1 + lo} ^ ° ^{(c l 'Dimetn} yl ^{formamide); NMR} spectrum (in dimethylsilfoxide dg): £, 8.72 and 8.60 (each 5-proton singlet can be transferred to isopropylidene protons, * £ - 1.5-2.8 (5-proton multiplet can be transferred to benzoyl protons ).

Example 5

Synthesis of 2 ⁿ -0 ~ benzoyl-penta-N-ethoxycarbonyl-4 ", 6" -0-isopropylidene-3 ', 4 ^f -di-0-mesylkanamycin B (in the general formula II, R _- OC ₂ H ₅ , Y = CH (CH- ^) ₂ , W = SO ₂ CH-, Z at C-2 "is COCgH ₅ and Z at C-5 is HJj.

1.18 g of the kanamycin B derivative prepared according to Example 4 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 ^ SO 2 Cl).} The solution was evaporated to a third of the original volume and the solution thus prepared was poured into water. The precipitates formed were filtered off and washed with water. 1.32 g of a pesticide with a melting point were obtained. point of 198 ⁰ C. jut) _D + Io7 ° (c 1.5 dimethyl formamide j ^! NMR spectrum (in pyridine d ₅ ): Z- 6.60 and 6.45 (each 3-proton singlet can be transferred to mesyl- protons).

1 098JSf / 18 50

Example 6

Synthesis of 2 ⁿ -0-benzoyl-3 ^l , 4 ^l -dideoxy-3 ^l , 4 ^l -didehydro ~ penta-N-ethoxycarbonyl-4 ¹¹ , 6 "-O-isopropylidene-kanamycin B.

11 g of sodium iodide and 5 g of zinc dust were added to a solution consisting of l, og of the kanamycin B derivative prepared according to Example 5 and 20 ml of dimethylformamide, and the mixture was heated to 95 ° C. for one hour with vigorous stirring. 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 and dried over sodium sulfate. After evaporation, 9Λ0 g of a solid were obtained. The pesticide was treated with hot ethyl acetate and 31ο 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., (et) ^ ² + 36 ° (c 0.4, dimethylformamide); NMR spectrum (in pyridine-dc): -7 ^ 4, o6 (2-proton in a somewhat broadened singlet, H-2 ¹ and 4 ^f ).

Example 7

Synthesis of 2 ^w -0-benzoyl-3 ^l , 4 ^l -dideoxy-penta-N-ethoxycarbonyl-4 ", 6" -O-isopropylidinkanemycin B.

1.15 g of the Kanamycin B derivative prepared according to Example 6, which in a mixture of 45 ml of p-dioxane 25 ml Methanol and 25 ml of water had been dissolved, was with hydrogen under 2 atmospheric pressure and in the presence of 0.3 g of platinum oxide that had previously been activated, filtered at room temperature for 5 hours. The so-

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The solution provided was filtered and evaporated. This gave l, lo g of a colorless powder having a melting point 254-256 ⁰ C, (POR) ^{2 "1" 87} ° ^(c ° ^{'2 Dirnetn} y ^lformamid i NMR "spectrum (in pyridine-d ^): ~ [ r 7.8 -8.5 (4 proton wide signal, H-2 ^f , 4 ^f ).

Example 8

Synthesis of 2 "-0-benzoyl-3 ^f , 4'-dideoxy-penta-N-ethoxycarbonylkanamycin B.

980 mg of the kanamycin B derivative prepared according to Example 7 were dissolved in a mixture of 8 ml of acetic acid and 5 ml of water. The solution was heated at 100 ° C for 5 minutes. The resulting precipitates were filtered and dried, and 75 ° g of a substance with a melting point of 247-248 ° C. were obtained. C &) d 3 89 ° ( _c 2.7, dimethylformamide). Found: C, 52.41; H, 6.54; N, 7.48 #. Calculated for C ₄₀ H ₆₁ N ₅ O ₁₀ : C, 52.45; H, 6.71; N,

Example 9

Synthesis of 3 ^f »4 ^f -dideoxykanamycin B.

150 g of the kanamycin B derivative prepared according to Example 8 and 2.3 g of barium hydroxide octahydrate were added to a mixture of 5 ml of water and 4 ml of p-dioxane. The so The resulting mixture was heated in a boiling water bath for 8 hours. Then carbon dioxide was introduced and the resulting precipitates were removed by centrifugation. The supernatant layer was isolated and evaporated. The residue obtained was dissolved in water and the solution was prepared according to Pil-

1 0 98 86} ² 1 8 5 0

tration on a loaded with Amberlite IRC-50 (H ⁺ form) with O ₅ I - 0.4 N ammonium hydroxide. The fraction containing 3 ', 4 ¹ - dideoxykanamycin B was concentrated. By adding acetone, 47 mg of a crystalline substance were obtained. (cC) ° + 130 ° (c 0.65, water. Found: C, 47.71; H, 8.38; N, $ 15.31. Calculated for C ₁₈ H ₅₇ N ₅ Og: C, 47, 88; H, 8.26; N, 15,

Example Io

Synthesis of penta-N-salicylidene-kanamycin B.

52o mg of salicylaldehyde were added to a suspension of 400 mg of Kanamycin B base in aqueous methanol (1: 8, 20 ml) added and the resulting solution was poured into water. The precipitates were filtered, dried and yielded 750 mg.

Example 11

Synthesis of 3 ^f , 4 '; 4 ^lt , 6 "-Di-0-cyclohexylidene-penta-N-salicyl-idenkanamycin B (in the general formula I ^!, R = C ₆ H ₄ (OH), X = Y = (CH ₂ ) ₅ C, Z = H).

7oo mg of the penta-N-salicylidene-kanamycin prepared according to Example Io B were dissolved in 12 ml of dry dimethylformamide and the solution was after the addition of 500 mg cyclohexanol dimethyl ketal and 3o mg anhydrous p-Tluenesulfonic acid heated at 50 ° C for one hour. Then a small amount of the solvent was removed by evaporation in vacuo. The solution made in this way was poured into water. The resulting precipitates were filtered, dried, and yielded 7I0 mg.

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Example 12

Synthesis of 2 "-0-Benzoyi-3 ^l , 4 ^f ; 4", 6 "-di-0-cyclohexylidenepenta-N-salicylidene-kanamycin B (in the general formula I, R = C / -Hji | (OH), X = Y = (CHo) CC * 12 = COC / rH, -)

59o mg of the kanamycin B derivative prepared according to Example 1 were dissolved in 10 ml of pyridine, and 120 mg of benzoyl chloride was added to the solution. One left an hour stand at room temperature for a long time and then the solution was poured into water. The resulting precipitation were filtered off, dried and a yield of 612 mg was obtained.

example

Synthesis of 2 "-O-Benzoyl-4", 6 "-O-cyclohexylidene-penta-N-ethoxycarbonylkanamycin B.

A mixture of 600 mg of the kanamycin B derivative prepared according to Example 12 and aqueous acetic acid ( 1: 3 » 20 ml) was heated at 95 ° C. for 30 minutes. The reaction mixture was evaporated and the residue treated with ether. The ether-insoluble part was dissolved in aqueous acetone (ItI, 15 ml). After adding 400 mg of sodium carbonate, 27o mg of ftthoxycarbonyl chloride were added to the solution. 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 of a solid substance were obtained. (oQ _D ° + 98 ⁰ (c 1.5 dimethylformamide.) This substance could be converted into 3 ^X , V-dideoxykanamycin B by means of a number of similar processes which are described in Examples 5 to 9.

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Claims (1)

Claim Process for the preparation of 5 ^f , 4 • -Dideoxykanamycin B with kanamycin B as starting material, "thereby geke η η records that the amino groups and all or part of the hydroxyl groups that are not J5 ^f - and 4 ^ hydroxyl groups of kanamycin B are protected in a known manner, the 3 ¹ - and 4 ¹ -hydroxyl groups are sulfonated in order to obtain a derivative with j disulfonic ester groups, the 3 ', 4 ^f -disulfonic ester are removed in a conventional manner to give a J5 ¹ , ¹ * ¹ -unsaturated To obtain compound, the 3 *, 4 * -unsaturated compound is reduced and the remaining protective groups are removed. - 15 109886/1850