SU1258327A3 - Method of producing 7-substituted compounds of 9a-methoxy-mitosane - Google Patents

Description

sn

 one

The invention relates to uchyuk) new derivatives of mitosan, namely, 7-valued 9o-methoxylimitosan of the general formula

СН20С- КНг ОШз

where A is a group of the formula

1 2N3-s, where R, is hydrogen or -NCCH); Rj is hydrogen, CH, or groups of general formulas

.N-СНз

. f .m

 SNS

1 GH,

R is methyl or Rj, together with R and the nitrogen atom to which they are attached, form pyrrolidine or piperidine, possessing antitumor activity.

The aim of the invention is to search for new compounds of the mitomycin series with high pro-tumor activity, as well as used as intermediate products in

synthesis of other compounds possessing this activity.

The invention is illustrated in the following examples.

PRI me R t and

A. To a solution of 1 t of isopropyl formamidate hydrochloride in 2 ml of dimethylfrmamide (DO)), 2.1 mMdiisopropyl ethylamine are slowly added at 0 ° C in an az. Atmosphere. To the half-diluted solution is added dropwise / -trimethylsilyltnp chloroformate at

0

583271

The resulting clean solution is designated with solution Ai

B. A solution of 1 mmol of i-otomycin C in 5 ml of DMF is added to the suspension.

5 1.5 mmol of sodium hydride in 3 ml of DMF. The solution is stirred at room temperature for 20 minutes and cooled to (-40) - (- 50) ° C before adding the above prepared solution A. The solution is kept at 40 ° C for 1 hour, then allowed to warm to room temperature, after which a the v6-l8 h is diluted with methylene chloride and filtered. The solid residue obtained after evaporation of the filtrate is chromatographed on silica gel to give the amidino protected compound indicated in the title of the example.

C.A | the idio-protecting group of the previous intermediate is removed to give the unsubstituted amidine compound indicated in the title of the example.

PRI me R 2

five

0

five

OCONH2

thirty

OSSN

NH

A. To a solution of 1 mM isopropylformamidate hydrochloride in 2 ml of DMF, 21 mM diisopropyl methyl is slowly added under nitrogen at 0 ° C. Methyl iodide is added to the solution obtained. Polzgchenny solution is designated as solution B.

B. Using solution B instead of solution A, Example 1B is repeated.

to obtain the compound indicated in the title of the example.

PRI me R 3.

ShNz) 2 about

) I

НII 1

OCONH2 OSN

kn

A 0.5 m solution of M, N-dimethylchloromethylnilvinium chloride is obtained by 55 dropwise addition of 1.57 g oxalyl chloride (12.5 mmol) at 0 ° C to a solution of 915 mg (12.5 mmol) DMF ml of chloroform, and stirred for 30 minutes

1fi room temperature. Separately, a solution of 334 mg (1 kmol) mitomycin C in 5 ml of Daph is added to a suspension of 36 mg (1.5 mmol) of sodium hydride in 3 ml of DMF. The solution is stirred at room temperature for 20 minutes and cooled to (-40) - (-50) C after which the described solution of N, H-dimethylchloromethyleneimine chloride (3 ml, 1.5 mmol) is added. After 10 minutes of stirring at -40 ° C, 18 mg (0.75 mmol) of sodium hydride is added to the solution. The solution was held at -40 ° C for 1 hour and then diluted with methylene chloride and filtered. The residue obtained after evaporation of the filtrate was subjected to flash chromatography on silica gel (10% as eluant).

Extraction of the main green band gives 78 mg (43% relative to the returned mtomycin C) amorphous solid. Extraction of the 4n-1 band gives 150 mg of mitomycin C.

Example 4. 7 (1-Methyl-2- (H) - -pyridinylidene) -amino-9a-methoxy-mitosan

OCONH2

DOS

Bn

Sh

To a mixture of 242 mg (0.725 mmol) of mi-TOwnptHa C and 43.5 mg (1.81 mmol) of sodium hydride was added 4 ml of DMF. After 15 minutes of stirring at the natural temperature, 370 mg (1.45 mmol) of 2-chloro-1-methylpyridinium iodide are added. The solution is stirred for 1.5 hours, diluted with ethyl acetate (t OAc) and filtered. The residue obtained after evaporation of the filtrate is chromatographed on silica gel using 5% methanol in methylene chloride as the eluent. The minor product (12 mg) is Compound XIX. The main product (75 mg) purified by silica gel TLC using 10% methanol in methylene chloride to give 6 mg (2%) of the title compound.

12583274

PRI me R 5. 7- (Metstaminrmethylene) -amino1-9q-methoxymitosan

, r-OCONH2 OCH-j

H

12 mg (0.5 mmol) of sodium hydride in a nitrogen atmosphere are added to a solution of 167 mg (0.5 mmol) of Mitokin C in 2 ml of hexamethylphosphoramide. 19 mg (0.25 mol) of N-methyl formimidoyl chloride are added to the solution, stirred at room temperature for 10 minutes, and then 6 mg (0.25 mmol) of hydride and 9.5 mg (0.13 mmol) are added. N-ketilformamidoyl chloride. After stirring for 6-12 hours, the solution is diluted with ethyl acetate and filtered. Evaporation of the solvent followed by chromatographic purification of the residue yields the compound indicated in the title of the example.

PRI me R 6. 7- (1-Pyrrolone-inylnetylene) -amnno-9a-methoxymitozan

O G-OsoYANG

OSSN

A 0.5 m solution of pyrrolidinyl chloromethyleneimine chloride is obtained by adding dropwise m 3.17 g (25 mmol) of oxalyl chloride with to a solution of 2.48 g (25 mmol) of 1-formyl pyrrolide in 50 ml of chloroform, followed by stirring at room temperature. round for 30 min. Separately, 24 mg (1 mmol) of sodium hydride was added to a solution of 334 mg (1 mmol) mitomycin C in 3 ml of 1-foropylpiperidol under a nitrogen atmosphere. After 20 minutes of stirring at room temperature, the solution is cooled to (-40) - (- 50) ° C and 1 ml (0.5 mmol) of the above prepared solution of imimium salt is added. At this mixture, 12 mg (0.5 mmol) of sodium hydride, 0.5 ml (0.25 mmol) of a solution of iminium salt, 6 mg (0.25 mmol) of sodium hydride, 0 , 25 ml

(0.125 mmol) of a solution of imineic co-gly, 3 mg (0.125 mmol b) of sodium hydride and O, t25 MP (0.063 mmol) of a solution of iminium salt. After 30 minutes of stirring, the mixture is warmed to room temperature. It is then diluted with ethyl acetate and the inorganic salt is filtered off. The residue obtained after evaporation of the solvent was chromatographed on silica gel by thin layer chromatography using 10% methanol in methylene chloride. Extraction of the green band gives 120 Ib (t5% by weight) of the compound indicated in the title of the example.

Example 7. 7 - {, L-Methyl-H (methylamino) -methyl-amino-9l-methoxy-mitosan

with aluminum. The column is eluted first with methylene chloride, and then with 2% -Hbw (v / v) methanol in stocky methylene. A green fraction is obtained, consisting of the desired product, which is further purified by chromatography on aluminum using methylene chloride containing 10% (v / v) methanol, yield 20 mg (5%).

Found,% C 52.68; H 6.21; N 18.15

Calculated

N 8.55

1-1 / 4 K.jO%: C 53.03; H 6.34;

PRI me R 9. 7- (1,3-Dimethyl-tetrahydropyrimidinylidene) -amino-9c (-methoxy-timitosan

 9%

i i

ABOUT

c

Ш2СЮЪ1Н2

mass

5-NH

Example 5 is repeated, replacing mito-mit C 9a-methoxy-7- (H-methylamino) mitosan in the same molecular amount.

PRI me R 8. 7- (1,3-Dimethyl 2-imidazoline) -9 (1,-methoxyimitosan

ABOUT

/ iHaOCNH

OSSN

sh

0.34 g (1 mmol) of mitomycin c is dissolved in 5 ml of 1,3-dimethyl-2-α-imdzazolidone and O is added to the solution, i g of sodium hydride (50% in oil 2.08 mmol) at room temperature. The mixture was held at room temperature for 20 minutes and then cooled in an ice-cold salt bath to -15 ° C. At this temperature, the mixture was held for 10 minutes, and then 0.65 g (2 mmol) 2-chlorine was added to it-1 , 3-dimethyl-4,5-dihydro- (3N) -imidazolimium chloride. After that, the mixture was again kept at -15 s for 1 h and then diluted with ethyl acetate. and chromatographic on a column with

Sodium hydride (50% dispersion in oil) in an amount of 200 mg (4.2 mmol) was added under a nitrogen atmosphere to a solution of 680 mg (2 mmol) mito in dyne in 8 ml of 1,3-dimethyl-3,4 , 5,6-tetragndro (1H, 3N) -2-pyrimidinone. The mixture is dried at room temperature for 20 minutes and cooled to -25 ° C. Then, 0.73 g (4 mmol) of 2-chloro-1,3-dimethyl -2,3,4,5-t of tetrahydropyrivium chloride was added to it, after which the mixture was added for 3 hours. Then it was diluted with ethyl acetate and 2 ml of methanol and, without further processing, applied to a chromatographic column with dry aluminum and zoned with first methylene chloride and then with a 2% (v / v) mixture of methanol / chlorine methylene to give the desired product 0.35 g (39.5%), m.p.

C, 54.78; H 6.18;

NgO

138- | 40 C.

Found, Z; N 18.21

,

&) 1 number,%: C 54.65; H 6.33; N 18,2t

Example 10. 7- (Tetramethyl-dyagnetillek) -amino-9a-metoksimitotan

N

.r-oeo-NH2

OCH H

425 MG (1.42 mmol)} "1 tomycin C is mixed with 50% sodium hydride dispersion in oil (85.3 mg) and 4 ml of dimethylformamide is added to the mixture, stirred at room temperature under argon atmosphere to and then cooled to -35 ° C. 289 mg (2.13 mmol) of tetramethyl chloroformamidium chloride are added and the mixture is allowed to warm to 5 ° C for 2 hours. Then dry ice is added to the mixture to stop the reaction and the solvent is distilled off under reduced pressure. The residue is chromatographed on a column of aluminum (100 g) using 3% (v / v) methanol in methylene chloride as eluent. This substance further purifies those on aluminum (5% (v / v) meta-iol in sulfuric methylene) to obtain two fractions of 17 mg and 76 mg in weight. The second fraction is crystallized from acetone-ether from the target product, t, sh1, yield 12%.

Found,%: C 54.92; H 6.53; N 19.29

With "ONG8. MBO

Calculated,%: C 55,54; H 6.53; N 19.43

Example 11, 7- (1-Piperidinsh1-methylene) -amino-9a-methoxymitosan

by creeping 193-195 С,

ABOUT

about

CH OCKHi OCH-i

A 0.5 M solution of piperidinyl chloromethylenimine chloride is obtained by adding dropwise 380 mg (3 mmol) of oxalyl chloride and 6 ml of chloroform containing 0.34 g (3 mmol) of 1-formylpiperidine. Separately, sodium hydride (30% dispersion in oil) in an amount of 96 mg (2 mmol) is added to a solution of 334 mg (1 mmol) mitomycin C in 3 ml of 1-formylpiperidine under a nitrogen atmosphere. After 15 minutes

eight

ten

15

20

25

thirty

five

0

five

After stirring at room temperature, the solution is cooled to and 4 ml (2 mmol) of the solution of the iiiiea salt prepared as described above are added. The reaction mixture is kept at -25 ° C for 1 hour and then cooled by the addition of dry ice. After methanol (ml) is added, the mixture is absorbed with neutral aluminum, which is then applied to an aluminum column (30 g). The column is eluted first with methylene chloride and then 3% (v / v) methanol in methylene chloride with obtaining 360 mg (84%) of the compound indicated in the title of the example, mp. 68-70 С,

Found: C 55.57; H 6.21; N 15.91

 1-1 / 4

Calculated,%: C 55.80; H 6.58; N 15.49

Antitumor activity on. . Relationship to P-388 Murine Leukemia.

The table presents the results of laboratory research on CqP | Female mice that were intraperitoneally implanted with a tumor inoculum containing 10 ° ascites P-388 iBurine leukemia cells and administered various doses of a test compound of the indicated formula I or mitomycin C, Group of. She received a dose of dinane on the 17th day after infection. In each series of experiments, a group of 10 mice were inoculated, which were injected with saline. The groups treated with mit (C) were a half control. In each group of bathes at 30 days, the survival time was noted at the bottom and the number of SURVIVAL individuals was recorded. The smaller one was weighed before the administration of the preparation and again on the 6th Weight change was taken as a measure of toxicity of the substance. Mice weighing 20 g each were taken in the experiment, and weight loss up to -2 g was not considered excessive. The results were expressed in percentages of T / C load (treated / ctmtrol) ., which is the ratio of the survival time of the group treated with prepar atom, by the time of the survival of the control group treated with saline, which was taken as 100%, the animals that were injected with saline usually died no later than 99125832710

: th days Thus, the dose causing the OTHocHTenbHiaH vaccinating activity to a minimum effect (125% T / C) is likely to be effective. The comparison of the invention according to the invention in comparison is not given in the table, the average is given with a heart rate of C. The minimum effect is the weight value per mouse at maximally considered 12% T / C. Minimally effective and minimally effective doses.

Suppression of P-388 Murine Leukemia

Note: Numbers in parentheses represent

values obtained by administering momitin C as a positive control.

The proposed compounds were tested for antitumor activity infected with B 16 melanoma. The experiments were carried out on DFt microscopes, which underwent tumor implant intraocularly for 60 days. For each group of 10 minutes, the shaving time was determined. T control animals inoculated in the same way as the experimental animals and injected: with a solvent without a drug, the survival time was 21 days. The T / C ratio (%) was used to determine the maximum and minimum effective doses of each compound. Such a dose was taken as the minimum effective, which caused a 125% T / C effect. For each dose, the tested Y compound was intraperitoneally injected into the test animals on days 1.5 and 9. The average weight change per day, noted at the maximum and minimum effective doses, was used as a measure of toxicity. A weight loss of 2 g for a 20 g mouse was not considered excessive.

but also the minimum effective doses.

The compounds of Examples 10 and 9 were tested for anticancer activity with respect to B 16 Chgpe nbpa oi after subcutaneous administration of tumor tumor material and intra drug administration. The drug administration schedule and the withdrawal time (the study was carried out for 40 days) were determined by the known method. The change in weight was measured on day 12-6. The maximum effective dose of compound 1 mg / kg caused an effect of 156% (T / C) and a weight gain of 1.5 g. In groups of 6 animals per day, 3 mice survived for 40 days . The minimum effective dose was 0.25 mg / kg, at which on the 12th day the weight change was 1.0 g. For the compound of Example 9, the maximum effective dose was 8 mg / kg with an effect of 177% T / C and a weight change of 0 , 6, the minimum effective dose is 4 mg / kg With a weight change of 0.8. In the same experiment, the maximum effective dose of mitomycin C is 3 mg / kg with an effect of 195% T / C and a weight change of 0.5, the minimum effect is 123832712

The dose of mitomycin C is not determined by the antitumor activity, you divided it in animal experiments,

Thus, the data confirms, as well as reduced toxicity, for the compounds obtained are highly compared with mitomycin C,

Compiled by I. Bocharova Editor M. Kelemesh Tehred M. Khodanich Proofreader-A. Obruchar

Order 5043/60 Circulation 379Subscription

VNIIPI USSR State Committee

for inventions and discoveries 113035, Moscow, Zh-35, Raushsk nab., 4/5

Production and printing company, Uzhgorod, Projecto st., 4

Claims (1)

1. METHOD FOR PRODUCING 7-SUBSTITUTED 9a-METHOXYMITOSANE of the general formula BUT CH ₃ ABOUT I) / CH ₂ OS-No. ₂ 0CH L> NH where A is a group of the general formula where R _t is hydrogen or —N (CH ₃ ) ₁ ; - hydrogen, CH * or groups of general formulas R e - methyl or R _g together with R _e and the nitrogen atom · to which they are attached form pyrrolidine or piperidine, characterized in that 1.0-1.5 act on a solution of methomycin C in an organic solvent such as dimethylformamide with a molar amount of sodium hydride and subsequent processing to form the anionic form of metomycin With an electrophilic reagent selected from iminoether, halomethylene imine halide or iminohalide salt * SU <-> 1258327 AZ 1258327 2