DE1918316C3 - Mitomycindenvates and process for their preparation - Google Patents

Description

in which X is a methoxy or amine group and either R is a hydrogen atom and Z is a hydrogen atom or a methyl group or R is Acetyl group and Z denotes a methyl or acetyl group.

2. Process for the preparation of the mitomycin derivatives of the general formula I according to claim 1, characterized in that either

a) a compound of the general formula

OCH ₃

CH ₂ OCONH ₂

(II)

The invention relates to mitomycin derivatives in general formula

CH ₂ OR

wherein Z is a hydrogen atom or a methyl group. with a 0.2 to 1 abnormal aqueous or aqueous alcoholic alkali metal hydroxide solution for 16 to 2 hours and the obtained compound of the general formula

O
HO II CH ₂ OH

(III)

can react with diazomethane or

b) a compound of the general formula II in which Z is a hydrogen atom or a methyl group means, reacts in an organic solvent with an alkali metal alcoholate or

c) a compound of the general formula II, in which Z is a hydrogen atom, in an organic solvent is reacted with lithium aluminum hydride and the product obtained with potassium nitrosodisulfonate, Air or ferric chloride is oxidized

and optionally then a compound of the general formula I obtained, wherein X is a methoxy group, converted into the amino compound with ammonia and /

3 ° in which X is a methoxy or amino group and either R is a hydrogen atom and Z is a hydrogen atom or a methyl group or R an acetyl group and Z a methyl or acetyl group means and process for their preparation.

The new compounds are referred to below as decarbamylmitosans.

The Decarbamylmitosane of the general formula I are prepared by either

a) a compound of the general formula

CH ₂ OCONH ₂

in which Z is a hydrogen atom or a methyl group, with a 0.2 to 1 onormal aqueous or aqueous alcoholic alkali metal hydroxide solution for 16 to 2 hours and the resulting Compound of the general formula

HO

(III)

NZ

can react with diazomethane or

b) a compound of the general formula H, in which Z is a hydrogen atom or a methyl group means, reacts in an organic solvent with an alkali metal alcoholate or

c) a compound of the general formula II. wherein Z represents a hydrogen atom in an organic solvent with lithium aluminum hydride reacts and the product obtained with potassium nitrosodisulfonate, air or ferric chloride oxidized

and optionally then a compound of the general formula I obtained in which X is a methoxy group, converted into the amino compound with ammonia and / or one obtained A compound of the general formula I in which R is a hydrogen atom and Z is a hydrogen atom or is a methyl group, with acetic anhydride or an acetyl halide in the presence an organic base or with acetic acid in the presence of dicyclohexylcarbodiimide.

In well-known Japanese patent application 22 380/67 there is a method of manufacturing a Milomycins of the formula

HO

CH ₂ OCONH ₂
OCH ₃

NZ

wherein Z is a hydrogen atom or a methyl group, by reacting a mitomycin of the general Formula II described with alkali hydroxide. If you work with alkali hydroxide in higher concentration or using a longer reaction time than in the aforementioned Japanese patent application according to procedure a), the carbamyl group is also split off.

In the methylation of the compounds of the general Formula III with diazomethane is expedient of dissolving these compounds first Dry ice was added in excess of the amount required to neutralize the solution. Then it will be the solution is concentrated in vacuo, and the pH of the residue is increased to 3.0 by the addition of acid 4.0 set. The mixture thus obtained is extracted with ethyl acetate and the extract is extracted with a solution of diazomethane in ether. Since after the methylation there is still some mitomycin of the general formula II, in which X is the methoxy group, is present, the reaction mixture must according to the Methylation, for example by silica gel? Chromatography.

The preparation of the compounds of general formula I by reacting a mitomycin of general formula II in an organic solvent with an alkali metal alcoholate according to the with b) designated procedure has the advantage that the product is in high yield in a single stage Implementation, d. H. without the amino or methoxy group in the 7-position of the mitomycin of the general Formula II is converted into a hydroxyl group is obtained. It can be an alcoholate of a primary, secondary or tertiary alcohol in an organic solvent, for example an alcohol, Tetrahydrofuran, dioxane, dimethylformamide or benzene can be used in solution. The implementation takes place preferably at room temperature. When using a solution, the reaction time is Sodium methoxide in a mixture of methanol and benzene preferably about 8 hours and at Use of a 2N sodium isopropoxide solution in isopropanol, preferably 3 levels. To neutralization

tion of excess alcoholate is added dry ice in excess, and the reaction mixture is then evaporated in vacuo. From the return

• the product is extracted with acetone. The extract is concentrated and, for example, by

Purified silica gel chromatography to give the crystalline product.

Procedure c) is expediently carried out in such a way that the mitomycin of the general Formula II in a suitable solvent, for example tetrahydrofuran, ether, dioxane or Dimethylformamide, dissolved and the solution then lithium aluminum hydride in excess over the for Reduction of the carbamyl groups required amount is added. The reaction can take place at one temperature between that achieved by cooling with ice and the boiling temperature and is expediently carried out at room temperature. The reaction time can be between a few minutes and vary a few hours. After the completion of the reaction, residual lithium aluminum hydride becomes and the resulting lithium aluminum complex compound is decomposed with aqueous ethyl acetate. Then the mixture is oxidized with potassium nitrosodisulfonate, air or ferric chloride. By extraction the reaction mixture with an organic solvent and purification of the product Customary measures, for example silica gel chromatography, give the desired product. Decarbamylmitosans thus obtained, wherein X is a methoxy group, can then if desired can still be reacted with ammonia to convert the methoxy group in the 7-position into an amino group to convict.

The compounds of general formula I are antibiotics.

The decarbamylmitomycins of the general formula I, in which R is a hydrogen atom and Z is a hydrogen atom or a methyl group, generally have a lower antibiotic effect than the other mitomycins of the general formula I, but are also still effective. They are also valuable intermediate compounds for the production of other antibiotics, for example the other mitomycins of the formula 1. The antibiotic effect and the toxicity of some decarbamylmitosans according to the invention and the known mitomycin C are shown in Table I. In this table, the minimum concentration of decarbamylmitosans in y / ml of the solution required to inhibit the growth of pathogenic bacteria, determined by the agar plate dilution method, and the toxicity of the compound given as LD ₅₀ when administered intravenously to mice.

H 5 χ Z H 1 918 316 Sarcina lulea S. 6th Toxicity, CO -CH ₃ CO -CHj ■ J intravenous,
in mice H CH ₃ ATCC 10011 LD ₅₀ CO-CH ₃ CH ₃ O CH ₃ Table I. 1.58 Pseudomonas
aeruginosa mg / kg R. H CHjO H - Escherichia coli 10 CO-CH ₃ CHjO CO -CHj Bacillus
subtilis 3.125 ATCC 15246 20th H CHjO CH ₃ - ATCC 14948 3.125 > 100 CO-CH ₃ NH ₂ CH ₃ ATCC 6633 3.125 0.195 - 40 NH ₂ C. 0.78 - 0.048 6.25 > 33.8 NH ₂ 0.043 0.195 6.25 - 75 NH ₂ 3.125 - 0.78 1.56 > 100 Mitomycin 0.04 0.098 0.781 - > 100 0.195 1.562 > 25 9 0.05 > 25 - 3.125 0.78 1.56 0.048 1.56 0.049

The following examples illustrate the invention.

example 1

500 mg of the mitomycin of the general formula II, in which X is an amino group and Z is a hydrogen atom, were dissolved in 100 ecm of 0.2 n sodium hydroxide solution in a mixture of 50% methanol and 50% water. The solution was allowed to stand at 30 ° C. for 16 hours, whereupon it turned bluish purple in color. Then the pH of the solution was adjusted to 8.0 with dry ice. The solution was evaporated in vacuo at a temperature above 4O ⁰ C to a powder. 5 ml of water was added to the powder and the pH of the mixture was adjusted to 4.0 with hydrochloric acid. Then the solution was extracted three times with ethyl acetate. After dehydration with sodium sulfate, an excess solution of diazomethane in ether was added to the extract, and the mixture was allowed to stand for about 30 minutes and then dried in vacuo. The reddish purple residue was dissolved in chloroform and subjected to adsorption chromatography on silica gel. By developing with a mixture of acetone and chloroform in a ratio of 1: 1, adsorption bands formed. The fractions of the first eluate were concentrated to give an amorphous reddish purple powder. This powder was identified by infrared analysis (Fig. 1) as the decarbamylmitosan of the general formula I in which X is a methoxy group and R and Z are hydrogen atoms. The for the

C = O proportion of the group

Il

- CH ₂ OCNH ₂

characteristic absorption band, ie at 1700 cm " ¹ , had disappeared; yield 2%; no clear melting point. The starting mitomycin appeared in the second eluate.

Example 2

Following the procedure of Example 1 were from the mitomycin of the general formula II in which X

is a methoxy group and Z is a methyl group, reddish purple needle-like crystals were obtained, which were identified as the decarbamylmitosan of the general formula I, in which X is a methoxy group, R is a hydrogen atom and Z is a methyl group. The infrared spectrum is shown in Figure 2; Yield 1.5%; F. 180 ⁰ C.

Example 3

Following the procedure of Example 1 were from the mitomycin of the general formula II in which X is an amino group and Z is a methyl radical, reddish purple needle-like crystals obtained as the decarbamylmitosan of the general formula I, in which X is a methoxy group and R is a hydrogen atom and Z is a methyl group. The infrared spectrum is shown in FIG. 2 shown; Yield 20%; F. 180 ° C.

Example 4

The decarbamylmitosan of the general formula I obtained in Example 1, in which X is a methoxy group and R and Z are hydrogen atoms, was dissolved in methanol, and excess ammonia was added. The mixture was left to stand at room temperature for 1 hour. The bluish purple one obtained Solution was evaporated in vacuo to give bluish purple needle-like crystals, as the decarbamylmitomycin of the general formula I, in which X is an amino group and R and Z Are hydrogen atoms have been identified. The infrared spectrum is shown in FIG. 3 shown; Yield 85%: F. over 300 ° C.

Example 5

Following the procedure of Example 4, the decarbamylmitosan obtained in Example 2 became the general Formally, in which X is a methoxy group. R is hydrogen and Z is methyl. bluish purple crystals obtained as the decarbamylmitosan of the general formula I in the X is an amino group, R is a hydrogen atom, and Z is a methyl group. The Infrared spectrum is shown in Figure 4; Yield 90%; F. 165 to 167 ° C.

Example 6

100 mg of the mitomycin of the general formula II, in which X is a methoxy group and Z is a hydrogen atom, were dissolved in 10 ml of anhydrous tetrahydrofuran, and 80 mg of lithium aluminum hydride were added to the solution. The reaction was allowed to proceed with stirring at room temperature for 1 hour. Aqueous ethyl acetate was then added to the reaction mixture in order to decompose excess lithium aluminum hydride and the complex lithium aluminum compound. Immediately thereafter, the solution was poured into a solution of 500 mg of potassium nitrosodisulfonate in a mixture of 27 ml water and 14 ml of V6 ^{norma 's} primary potassium phosphate cast. The color of the latter solution immediately turned bluish purple in color. The bluish purple solution was extracted with chloroform, and the extracted solution was dried with anhydrous sodium sulfate. The purification was carried out by means of a chromatographic column on silica gel and using a mixture of chloroform and acetone in a ratio of 7: 3 as the eluent. First, reddish purple fractions containing the decarbamylmitosan of the general formula I, in which X is a methoxy group and R and Z are hydrogen atoms, were eluted. The subsequently eluted reddish purple fractions contained the starting mitomycin. The infrared spectrum is shown in FIG. 1 shown; Yield 5%; no clear melting point.

Example 7

The procedure of Example 6 was repeated using 100 mg of mitomycin of the general type Formula II, in which X is an amino group and Z is a hydrogen atom, as a starting material. First, purple-colored fractions containing the decarbamylmitosan of the general formula I in the X is an amino group and R and Z are hydrogen atoms, eluted. The subsequently eluted purple fractions contained the starting mitomycin. The infrared spectrum is in FIG. 3 shown; Yield 7%; F. over 300 ° C.

Example 8

500 mg of mitomycin of the general formula II, in which X is an amino group and Z is a hydrogen atom is, were in 170 ml of a 2 normal methanolic Solution of sodium methoxide dissolved, and the solution was left with stirring at room temperature for 24 hours. The greenish-blue solution obtained was mixed with dry ice until the greenish-blue color turned purple. The solution was at evaporated to dryness in vacuo at a temperature above 40 ° C., a bluish-green solid being obtained would. This material was extracted five times with 60 ml of acetone. The extracts were combined and evaporated to dryness, a blackish-purple paste-like residue being obtained would. This residue was dissolved in a 1: 2 mixture of chloroform and acetone and adsorbed on a silica gel column. By developing with a mixture of acetone and 1: 1 chloroform became a purple color Adsorption band obtained. The eluaf of this adsorption band was evaporated to dryness, dissolved in a small amount of methanol and left to stand at room temperature, turning purple colored needle-like crystals formed. This product was known as the decarbamylmitosan of the general Formula I, in which X is an amino group and R un <Z are hydrogen atoms, identified. In the infrared spectrum was for the

C = O proportion of the group

- CH ₂ OCNH ₂

characteristic absorption band,. ie at 1700 cm " ¹ , disappeared. The infrared spectrum is shown in FIG. 3; yield 55%; F. over 300 ^° C.

Example 9

^r

Following the procedure of Examples, purple needle-like crystals were obtained from mitomycin of the general formula II, in which X is an amino group and Z is a methyl radical. This product has been identified as the decarbamylmitosan of general formula I in which X is amino, R is hydrogen and Z is methyl; M.p. 165 to 167 ° C. The infrared spectrum is shown in Figure A ; Yield 70%.

. Example 10

500 mg of the mitomycin of the general formula II. In which X is an amino group and Z is a hydrogen atom, were added to a mixture of 50 ml of 2N sodium isopropoxide and 85 ml of isopropanol. The resulting mixture was stirred at room temperature for 3 hours. The resulting greenish-blue solution turned purple on standing. This solution was evaporated in vacuo at a temperature above 4O ⁰ C to dryness, the residue obtained as a blue-gray solid material was extracted five times with 60 ml acetone. The extracts were evaporated to give a blackish purple paste-like residue. This residue was dissolved in a mixture of chloroform and acetone in a ratio of 1: 2 and adsorbed on a silica gel column. A purple adsorption band was obtained by developing with a 1: 1 mixture of acetone and chloroform.

The main eluate was evaporated to dryness, dissolved in a small amount of methanol and allowed to stand at room temperature. Purple needle-like crystals formed. This product was identified as the decarbamylmitosan of general formula I in which X is an amino group, R is a hydrogen atom and Z is a hydrogen atom. The infrared spectrum is shown in Fig. 3. Yield 65%; F. over 300 ⁰ C. '

Example 11

500 mg of the mitomycin used as a starting material in Example 10 was dissolved in a mixture of 85 ml of 4 normal methanolic sodium methoxide and 85 ml of benzene, and the solution was stirred at room temperature for 8 hours. The greenish-blue solution formed was mixed with dry ice and then turned purple. It was at a temperature above 40 ⁰ C in a vacuum

300 648/282

7 C

■ u

Evaporated to dryness, a bluish gray solid material being obtained as residue. This material was extracted five times with 60 ml of acetone. The extracts were evaporated to dryness to give a blackish purple pasty residue. The residue was dissolved in a mixture of chloroform and acetone in a ratio of 1: 2 and adsorbed on a silica gel column. A purple adsorption band was obtained by developing with a 1: 1 mixture of acetone and chloroform. The main eluate was evaporated to dryness and the residue was dissolved in a small amount of methanol and allowed to stand at room temperature. Purple needle-like crystals formed. These crystals were identified as the decarbamylmitosan obtained in Example 10. The infrared spectrum is shown in Figure 3; Yield 45%; F. over 300 ⁰ C.

Example 12

50 mg of the decarbamylmitosan of the general formula I, in which X is a methoxy group and R and Z Hydrogen atoms were dissolved in 0.5 ml of pyridine, and the solution was added 0.5 ml of acetic anhydride added. The mixture was allowed to stand at room temperature for about 1 hour and then poured into ice water added to decompose the acetic anhydride and extracted with ethyl acetate. The extract was shaken with a 10% aqueous sodium bicarbonate solution to separate the acetic acid. The ethyl acetate solution was dried with anhydrous sodium sulfate and evaporated to give 40 mg an amorphous powder were obtained, which as the decarbamylmitosan of the general formula I in the X is methoxy group and R and Z are acetyl groups. The infrared spectrum is in Fig. 5 is shown; Yield 63%; no clear melting point.

Example 13

50 mg of the decarbamylmitosan of the general formula I, in which X is an amino group and R and Z Are hydrogen atoms, were dissolved in 0.5 ml of pyridine and treated with 0.5 ml of acetic anhydride. the The solution was left to stand for about 1 hour and then poured into ice water and further allowed to stand to decompose the acetic anhydride. Then it was extracted with ethyl acetate. The extract was made with shaken a 10% aqueous sodium bicarbonate solution to separate the acetic acid. the Ethyl acetate solution was dried with anhydrous sodium sulfate and evaporated. You got that Decarbamylmitosan of general formula I, in which X is an amino group and R and Z are acetyl groups are. The infrared spectrum is shown in Figure 6; Yield 75%; oily substance.

Example 14

50 mg of the decarbamylmitosan of the general formula I, in which X is an amino group, R is a hydrogen atom and Z is a methyl group, were dissolved in 5 ml of dried tetrahydrofuran, and 1 ml of triethylamine and 0.5 ml of acetyl chloride were added to the solution and overnight left standing in a cold room. The triethylamine hydrochloride formed was filtered off and the filtrate was evaporated in vacuo. 40 mg of purple crystals were obtained, which were identified as the decarbamylmitosan of the general formula I, in which X is an amino group, R is an acetyl group and Z is a methyl group. The infrared spectrum is shown in FIG. 7 shown; Yield 70%; F. 172 to 174 ⁰ C.

Example 15

50 mg of the decarbamylmitosan of the general formula I in which X is a methoxy group and R is a water atom and Z is a methyl group was dissolved in 5 ml of tetrahydrofuran, and the solution became with 50 mg of dicyclohexylcarbodiimide, 0.5 ml of acetic acid and 0.5 ml of pyridine and 12 hours at Left to stand at room temperature. After the precipitated dicyclohexyl carbamide had been filtered off, the The filtrate was evaporated in vacuo and the residue was mixed with a small amount of a mixture added petroleum ether and ether to separate unreacted dicyclohexylcarbodiimide. the The residue was dissolved in ethyl acetate and shaken with a 10% sodium bicarbonate solution to Separate excess acetic acid, and then evaporated in vacuo. Reddish purple ones were obtained Crystals that are known as the decarbamylmitosan of the general formula I, in which X is a methoxy group, R is acetyl group and Z is methyl group. The infrared spectrum is in Fig. 8 shown; Yield 55%; M.p. 184 to 186 ° C

1 sheet of drawings

Claims (1)

Patent claims: 1. Mitomycin derivatives of the general formula O CH ₁ OR
OCH ₃ (D or a compound of the general formula I obtained in which R is a hydrogen atom and Z is a hydrogen atom or a methyl group, with acetic anhydride or one Acetyl halide in the presence of an organic base or with acetic acid in the presence of dicyclohexylcarbodiimide implemented.