CS268512B2 - Method of daunorubicine's and doxorubicine's new morpholine derivatives production - Google Patents

Abstract

Anthracycline derivs. of formula (I) are new. R is COMe or CO.CH2OH; B is CN or OH, or (with the H atom attached to the same C) is oxo; R'' is H and R' is OH, or B plus R' are together an oxygen bridge. Pref. B is CN or completes oxo or O bridge.

Description

(57) A process for the preparation of the novel ω-glutolins of daunorubicin and doxorubicin of formula (I) wherein R is -CO-CHo, -CHOH-CH 3, -CO-CH ₂ -OH and -СНОН-СН ₂ ОН, reacting the compound of formula II wherein R represents -CO-CH 3, -C 0 CH ₂ 0H, -CHOH-CH 3 or _-CH0H-CH2 0H, in a mixture of water and a polar organic media!

with 2.2 -oxydiacetaldehydem formula / CH ₂ -CH0 \

⁴ CH ₂ -CHO prepared in situ by cleavage of 1,4-anhydroerythritol in aqueous medium with sodium periodate at room temperature, the reaction is carried out in the presence of an alkali metal cyanoborohydride such as sodium or potassium cyanoborohydride and the crude reaction product obtained after morpholine ring closure neutral with alkaline fraction and neutral fractions are chromatographically purified to give pure product of formula I.

(I) (M o

T— <

VD 00 LO M

CO O

The invention relates to a process for the production of novel derivatives of daunorubicin and doxorubicin. These derivatives can be used as antitumor agents.

Doxorubicin (adrianycin) disclosed in U.S. Patent No. 3,590,028 to F. Arcaaone et al. Is probably the most effective anticancer agent currently in use. It is used concomitantly with daunorubicin to treat a variety of tumors as well as leukenia. However, some patients with these tumors, especially some serious types of tumors, such as colon and aelanoma tumors, do not respond favorably to this treatment. In addition, some patients with prolonged treatment suffer from incurable heart damage, which can lead to sarti. For this reason, it would be desirable to find analogous substances with a broad effect and lower toxicity to the heart muscle. Up to now some effective new analogs have been obtained, for example compounds active against murine leukemia Р388 / These are two lipophilic derivatives of AD32 and N, N-dibenzyl-idunorubicin, but they need to be used at high doses and do not affect the DNA in vitro, although DNA is probably the primary biological target of potent anthracycline derivatives. Most N-alkyl derivatives were also active against murine P388 leukemia, but this activity was not significantly different from that of doxorubicin or daunorubicin. Some of these derivatives were ineffective.

Known derivatives of doxorubicin and its anthracycline analogs are described in David w. Henry, "Adriamycin", ACS Symposium Serie, No. 30, Cancer Chemotherapy, American Chemical Society, pp. 15-57 (1976), and in Doxorubicin (Federico Arcamone, Academic Press, 1981.) Compound A032 has been described in US Patent Specification No. 4,035,566 of July 12, 1977.

3 ^< '3-deamino ^from - (4-morpholinyl) daunorubicin, disclosed in U.S. Pat.

301 277 from.17. Nov. 1981 (Edward M. Acton and Carol W. Mosher) is effective at 1/40 dose of doxorubicin but has essentially the same T / C, i.e. 166% compared to 160 for P388. This substance and its manufacture is also disclosed in Carol w. Mosher, Helen Y. Wu, Allan N. Fujiwara and Edward M. Acton "Enhanced Antitumor Properties of 3 '- (4-Morpholines1) and 3 ^Z - (4-Methoxy1-1-piperidines 1) Derivatives of з'-Oeaminodaunorubicin" , Э. Medicinal Chem., 25, pp. 18-24 (1982).

A general procedure for reductive alkylation to obtain novel semisynthetic anthracycline derivatives has been described in G. L. Tong, Η. Y. Wu, T.H. Smith and O. W. Hen ry “Adriamycines with Enhanced Efficacy and Reduced Cardiotoxicas”, □. Medicinal Chem., 22, 912-918 (1979).

A group of new daunorubicin and doxorubicin derivatives has now been found. These compounds can be represented by formula (I)

O-

where

R is -CO-CH3 or -CHOH-CH3 for daunorubicin derivatives or -COCH4 OH or -CHOH-CH3 OH for doxorubicin derivatives.

The compounds of the formula I can be obtained by reacting a daunorubicin or doxorubicin derivative of the formula

where

R is -CO-CH ₃ , -CO-CH ₂ OH, -CHOH-CH ₃ or -CHO-CH ₂ OH, in a mixture of water and a polar organic medium with a compound of formula

or with a suitable precursor thereof, wherein

Z is an oxygen atom, the reaction is carried out in the presence of a cyanoborohydride, for example an alkali metal cyanoborohydride, the resulting compounds are isolated and purified in a known manner. The compounds obtained are related to known antitumor agents, daunorubicin and doxorubicin (adriamycin), and are prepared from these agents in a chemical manner to form suitable derivatives of said agents which are also effective against tumors. These substances seem to combine both advantages, high antitumor efficacy and the possibility of using low doses. For this reason, side effects such as cardiotoxicity are also lower compared to known compounds.

The compounds of the present invention can be formulated into pharmaceutical compositions for the treatment of tumors by conventional methods:

The cyanmorpholinyl derivatives of daunorubicin and doxorubicin are shown in the following Table I.

Table I

Name of the compound co-ch ₃ choh-ch ₃ co-ch ₂ oh choh-ch ₂ oh

3 ^Z- deamino-3 ^z - (3'-cyano-4'-morpholinyl) daunorubicin

3 ^z -Deamino-3 ^z - (3-cyano-4-morpholinyl) -13-dihydrodaunorubicin

3 ^z -deamino-3 ^z - (3'-cyano-4'-morpholinyl) doxorubicin

3 ^Z -Deamino-3 ^z - (3 ** cyan-4-morpholinyl) -13-dihydrodoxorubicin

Four of these compounds are most preferred for their very good anti-tumor activity. For the following substances:

3 ^Z- deamino-з '- (3 * -cyan-4-morpholinyl) doxorubic in

3'-deamino-3 ^z - (3'-cyano-4-morpholinyl) -13-dihydrodoxorubicin,

3 ^z- deamino-3 ^z - (3-cyano-4-morpholinyl) daunorubicin and

3 ^of -deamino-3 '- (3-cyano-4'-sulfonolinyl) -13-dihydrodaunorubicin.

The first of these is the most preferred.

Process for making some preferred compounds. .

These compounds can be obtained by the following general method:

First, the commercially available daunorubicin or doxorubicin in the form of its acid addition salt is reacted under conditions of reductive alkylation with 2,2 ^z -oxodiacetaldehyde of the formula

O »CH

CH-O

CH ₂ - O - CH ₂

This alkylation yields a mixed product containing four major components.

For daunorubicin use those components are the following four compounds: 3 ^{out of} deamino-3 - (4'-raorfolinyl) daunorubicin, 3 ^{out of} deamino-3 - (4'-morfoliny1) -13-dihydrodaunorubicin, 3 ^T - deamino-3 ^from - (3-cyano-4'-morpholinyl) daunorubicin and

3 ^Z- Deamino-3 ^z - (3'-cyan-4'-morpholinyl) -13-dihydrodaunorubicin.

In the case of doxorubicin, these components are the following four compounds: 3 ^z -deamino-3 ^z - (4-morpholinyl) doxorubicin,

3 ^Z- deamino-3 ^z - (4-morpholinyl) -13-dihydrodoxorubin,

3 ^z -deamino-3 ^z - (3-cyano-4-morpholinyl) doxorubicin and

3 ^Z- Deamino-3 ^z - (3-cyano-4'-morpholinyl) -13-dihydrodoxorubicin.

-Oxodiacetaldehyd ^of 2.2 can be obtained by hydrolysis of 2,2-bis oxydiacetaldehyd (diethy1 acetal) in acidic medium. The starting material can be represented by the formula (Et-O) ₂ -CH

CH- (O-Et) ₂

O - СН _£

The process is carried out according to the method of Belgian Patent No. 655,436 (Field et al.), The same substance can also be obtained by cleavage of 1,4-anhydroerythritol of the formula

HO - CH ---- - CH - OH

CH ₂ - O -CH _£ according to literature procedures, Barry et al Carbohydrate Research, 7, 299 (1968) and Greenberg et al Carbohydrate Reeearch, 35, 195 (1974).

Reductive alkylation can be carried out using an excess of dialdehyde in a mixture of water and a polar organic medium, for example a mixture of water and acetonitrile, usually at pH 7 in the presence of a reducing agent such as an alkali metal cyanoborohydride such as sodium or potassium cyanoborohydride. This is a very easy to carry out reaction, which is usually completed within 1 hour or even less at room temperature. Reductive alkylation is explained in more detail in the following examples and is also used in U.S. Pat. No. 4,301,277 and described in □. Medicinal chem., 25, pp. 18-24 (1982). .

The reaction mixture may be worked up in any manner that provides the desired isolation and good separation of the reaction product. Extraction of the reaction product with acid is effective in extracting cyano-free materials from acid-insoluble and cyano-containing materials. The resulting material pairs can be separated into individual substances by chromatography, for example by column chromatography, thin layer chromatography or high pressure liquid chromatography.

The invention will be illustrated by the following examples.

Example 1 preparation, isolation and identification of 3 ^from -deamino-3 ^from - (3'-cyano-4'-morpholinyl) daunorubicin and 3 ^from -deamino-3- (3'-cyano-4'-morpholinyl) -13-dihydrodaunorubicin .

A solution of 31.5 g (0.30 mol) of 1,4-anhydroerythritol in 400 ml of water is stirred and cooled in a water / ice bath, and 53.0 g (0.25 mol) of periodate are added in small portions. sodium. The mixture was stirred at room temperature for 1 hour while a small amount of sodium iodate precipitated. The mixture was allowed to stand overnight and then adjusted to pH 7.3 with a few drops of saturated sodium bicarbonate solution and 50 mL of water was added to give a clear solution.

A solution of 9.2 g (16.3 mmol) of daunorubicin hydrochloride in 50 ml of a 1: 1 mixture of methyl cyanide and water was added to the dialdehyde solution and the mixture was stirred at room temperature. After stirring for 45 minutes, the mixture became cloudy and a small amount of solid formed. After adding 75 ml of methyl cyanide, only a portion of this solid dissolved. A solution of 2.02 g (32.6 mmol) of sodium cyanoborohydride in 6 ml of water was then added. After stirring for 1 hour, the mixture was diluted with water and extracted with 5 x 200 ml of chloroform, the solids 0.91 g which did not dissolve in any phase were collected by filtration.

After evaporation of chloroform the crude reaction product contains ^from 3 ^from deamino-3 - (4-morpholinyl) -daunorubic in 3 ^{out of} deamino-3 - (4'-morfoliny1) -i3 dihydrodaunorubic-ins ^from 3 -deamino -3 ^Z - (3-cyano-4'-morpholinyl) -daunorubicin and 3 ^z -deamino-3 ^z - (3'-cyano-4'-morpholinyl) -13-dihydrodaunorubicin. 5.4 g of the solid crude product were dissolved in 500 ml of a 9: 1 mixture of chloroform and methanol. The solution was washed with 0.01 N hydrochloric acid (3 x 100 ml), followed by 100 ml of water and 100 ml of dilute sodium bicarbonate. The organic phase is dried, evaporated to dryness and the residue is dried under vacuum at 1 mmHg at room temperature to give 5.10 g of a glassy product.

5.09 g of this glassy product is dissolved in 50 ml of 4: 1 methylene dichloride: methanol. To this solution is added dropwise methyl cyanide dropwise with stirring. A cloudy solution was obtained which was evaporated to dryness to give a semi-solid residue which was triturated in the dark with 200 ml of methyl cyanide. The insoluble material is separated and triturated a second time with 100 ml of methyl cyanide. The liquid phase of both processes contains the desired product. The liquid phases were combined and evaporated to give 2.23 g of semi-solid residue which is a crude mixture ^of 3-deamino ^from 3 - (3-cyano-4 ^E * morpholinyl) daunorublcinu and 3 ^from deamino-3 ^from - (3'-cyano-4'-morpholinyl) -13-dihydrodaunorubicin.

The semi-solid residue was then dissolved in 5 ml of methyldichloride and naness es onto a 3/1 x 59 cm column packed with silica gel (Mallinckrodt Silic AR CC-7) having an average particle size of 200 to 325 mesh in methylene chloride. The column is eluted with 500 µl of methylene chloride and then with methylene chloride / methanol first in a ratio of 1,500 ml and 99: 1, then 1,000 ml of the 98: 2 mixture, then 1,500 ml of the 97: 3 mixture and finally 500 10 ml fractions are collected, elution is monitored by thin layer chromatography. After collecting 2550 ml of the starting eluate, a 190 ml fraction was collected and evaporated to give 0.48 g of product. The primary component is determined by high pressure liquid chromatography and thin layer chromatography, indicating that it is a poorly pure 3'-deamino-3 ^from - (3'-cyano-4'-morpholinyl) daunorubinin.

• A sample of 0.35 g of this material was further purified first in the dark on five silica gel? 2 mm x 20 x 20 cm plates are washed twice with a 29: 1 mixture of methyl dichloride and methanol. A center band containing 65% by weight of the sample is cut out, the material is washed, the solution is filtered and evaporated. to dryness, together with other similarly purified materials, which are obtained in an amount of 0.18 g, after which the sample is purified on a column of 1.1 x 10 cm silica gel having an average particle size of 200-400 mesh. The column is eluted with methylene chloride / ethyl acetate, first with 30 ml of 80:20 mixture, 30 ml of 60:40 mixture, 30 ml of 40:60 mixture and 30 ml of 20:80 mixture and finally used. 175 ml of ethyl acetate. After collecting 162 ml of the starting eluate (2 ml fractions were monitored by thin-layer chromatography), 88 ml fractions were collected and evaporated to give 0.15 g of pure product.

Elemental analysis of this pure material as well as the NMR spectrum at 1360 MHz, ultraviolet, infrared and mass spectra showed that it was 3-deamino-3 '- (3'-cyano-4'-morpholinyl). daunorubif ci n.

The presence of this product as a di-isomeric mixture was shown by high pressure liquid chromatography and a NMR spectrum at 360 MHz. Both HPLC and Waters Radial-Pak C-18 α, δ M citrate buffer pH 4 and 60: 40 methyl cyanide at 2 ml / min showed two peaks at 9.6 and 10 respectively. The NMR spectrum at 360 MHz showed a double resonance for 6-OH, 11, OH, 1-H, 2-H, 3-H, 1'-H, 7-H, ОН 9, 10A-H, 14-H ₃ and H-6 ₃ - protons.

NMR spectrum at 360 MHz in CDC1 ₃ Vmax cT 13.99, 13.98 (2s, 6-OH);

13.25, 13.24 (2s, 11-OH), 8.02, 8.00 (2d, 1-H), 7.79, 7.77 (2t, 2-H), 7.40, 7 , 38 (2d, 3-H), 5.59, 5.56 (2d, 1'-H), 5.29, 5.26 (2bs, 6-H), 4.47, 4.34 (2s) 9-OH)

4.08 (S, OCH ₃ ), 3.97-4.07 (m, 2'BH), 3.92 (t, □-12Hz 3'-H), 3.74 (m, 2Ά-Η,

6'BH), 3.68 (bs, ^Z 4 H), 3.58 (t, □ ~ 12Hz, 6'AH), 3.20 (d, Э = 19Hz, H-10B), 2.91,

2.90 (2d, 0. 19HZ, 10A-H), 2.75-2.95 (m, 3'-H), 2.68 (m, 5'-H ₂ ), 2.43, 2, 42 (2s, 14-H ₃ ), 2.35 (m, 8B).

CS 268512 02

The Mallinckrodt Silic ARCC-7 silica column further eluted with a mixture of methylene dichloride and methanol after elution of 3 ' -deamino-3 &apos;-( 3 &apos; -cyano-4 &apos; -golpholinyl) daunorubicin. at a volume ratio of 90:10. After collecting 3,460 ml of the eluate, a 430 ml fraction is collected which contains 12.5% of the original material as a single compound. This fraction was evaporated and the compound was identified as impure 3 ^from -deamino-3 * - (3'-cyano-4'-morpholinyl) -13-dihydrodaunorubicin, which was further purified as described for its non-hydrogenated analogue to give yields substantially pure end product.

The spectrum at 360 MHz in CDCl ₃ is similar to the NMR spectrum for the first product of Example 1 except that the 14-H ₃ resonance (2.43, 2.42 2s) is shifted upward (1.32, 1.30 < -1 &gt;). J ”, 2d, □ 6.4 Hz). The mass spectrum in the form of the trimethylsilyl derivative (TMS) has m / e 957, [M (TMS) ₃ -HCN], 885 [m (TMS) ₄ -HCN4 | . In a thin layer chromatography on silica gel using a 19: 1 mixture of chloroform and methanol, _{Rf is} 0.8. '

Example 2

Preparation and isolation of 3'-deamino-3 '- (3'-cyano-4‘-morpholinyl1) doxorubicin and з'-deamino-3 * - (3‘-cyano-4'-morpholinyl) -13-dihydrodoxorubicin.

To a solution of 6.25 g (60.0 mmol) of 1,4-anhydroerhtritol OH OH in 75 ml of water cooled in a water bath at 15-20 ° C _9Θ, with stirring, add 6.42 g (30.0 mmol). sodium metaiodate, the resulting clear solution was stirred at room temperature for 17 hours. The pH of the solution was adjusted to 4.0-7.3 by the addition of sodium bicarbonate, then diluted with 75 mL of methyl cyanide with stirring. A precipitate formed. 0.126 g (2 mmol) of sodium cyanoborohydride in 5 ml of a 1: 1 mixture of methyl cyanide and water are then added with stirring; 1. To the resulting mixture was then added 1.16 g (2.0 mmol) of doxorubicin hydrochloride in 30 ml of a 1: 1 mixture of methyl cyanide and water. After 10 minutes, the reaction mixture was diluted with 50 ml of dilute sodium bicarbonate solution and then extracted three times. 50 ml of chloroform each. This crude extract contains 3 * -deamino-3 * - (4'-morpholinyl) doxorubicin, 3 * -deamino-3 * - (4'-morpholinyl) -13-dihydrodoxorubicin, 3 * -deamino-3 * - ( 3'-cyano-4'-morpholinyl) doxorubicin and 3'-deanino-3 '- (3'-cyano-4'-morpholinyl) -13-dihydrodoxorubicin. The extracts are combined and extracted 5 times with 25 ml each of 0.1 N acetic acid and then with water, washed with dilute sodium bicarbonate and saturated aqueous sodium chloride solution. The acidic aqueous phase is discarded. The chloroform phase is dried over sodium sulfate, filtered through Celite diatomaceous earth and evaporated, the residue is dissolved in 25 ml of chloroform and the solvent is again evaporated under vacuum at room temperature. 0.518 g of a dark red, glassy product is obtained in a yield of 40%.

0.424 g of this product was dissolved in 1.5 ml of methylene dichloride and applied to a 1.5 x 35.5 cm column packed with Bio-Sil A silica gel having a particle diameter of 200-400 mesh in methylene dichloride. The column is washed with 50 ml of methylene dichloride and then eluted with a mixture of methylene dichloride and methanol, first with 150 ml of a 99: 1 mixture, 150 ml of a 98: 2 mixture, 300 ml of a 97: 3 mixture, 100 ml of a mixture of 95: 5 and 300 mL of a 90: 100 mixture. First, 445 mL of the initial eluate was collected, then the 80 mL fraction was evaporated to give 0.217 g of product. This material was combined with 0.039 g of purified product from another preparation and the mixture was dissolved in 2 mL of methylene chloride, diluted with 10 mL of methanol and evaporated to dryness. This residue is triturated with 5 ml of methanol to give 0.218 g of 3-deamino-3- (3'-cyano-4'-morpholinyl) doxorubicin.

í i

CS 258512 B2

High pressure liquid chromatography and NMR analysis at 400 MHz indicated that the product was a diastereoisomeric compound. by high pressure liquid chromatography on a Waters Radial-pak C-18 column using 0.05 M citrate buffer pH 4 and methyl cyanide at 65:35 at a rate of 2 Bl / min, two close peaks at 14.4 and 15 can be detected The NMR spectrum of this material at 400 MHz has dual resonance for 1-H, 2-H, 3-H, γ-N, 7-H, 14-H ₂ , 9-OH, OCH 3. , 10A-H and б'-Н ^ -ргоtony. '

The NMR spectrum at 400 MHz in CDCl ₃ has maxima at 14.02 (s, 6-OH), 13.26 (s, 11-OH), 8.05, 8.04 (2d, 1-H), 7, 80, 7.79 (2t, 2-H), 7.41, 7.40 (2d, 3-H), 5.61, 5.57 (2d, G-N), 5.34, 5.30 , (2m, 7-H), 4.79, 4.78 (2s, 14-Hp, 4.54, 4.42 (2s, 9-OH), 4.11, 4.10, (2s, 0CH) ₃ ), 4.05 (m, 5 * -H), 3.97 (m, 2 * BH, 3 * -H, 6'BH), 3.71 (m, 4'-H, 6'-AH) ), 3.58 (t, 2Ά-Η), 3.30 (d, □ 19 Hz, 10B-H), 3.07, 3.06 (2d, 10A-H), 3.03. (» , З'-Н), 2.69 (m, 5'-H _2), 2.38 (m, H-8B), 2.22 (m, 8a-H), 1.84 (m, 2 ' -H ₂ ), 1.61 (s, H ₂ O), 1.40, 1.39 (2d, δ = 5.5 Hz, λ'-N ₃ ).

The ultraviolet absorption spectrum in methanol has maxima at 234 nm (4040,100), 252 (27,700), 289 (9,420), 478 (13,000), 495 (13,000), 495 (12,900), 530 (7,190).

Mass spectrum in the form of a trimethylsilyl (TMS) derivative, m / e 899 M (TMS) ₄ -HCN. Analysis for ^C 3 ^H 34 ^N ₂ 2 ° 12 ° * ^2H 2 Calculated: C, 56.97; H, 5.68; N, 4.15% Found: C, 57.07; H, 5.37; N, 4.17%.

Further elution of the column gave 0.041 g deamino-3 ^{y 3>} - ^{(3'-cyano-4-morpholinyl)} -13-yne dihydrodoxorub ic.

The ultraviolet spectrum in methanol has peaks at 234 nm (£ 37,400), 252 (28,000), 289 (9,390), 475 (12,700), 496 (12,800), 530 (7,410). Mass spectrum as trimethylsilyl derivative (TMS), m / e 985 M (TMS) ₅ -CH ₃ , 973 M (TMS) ₅ -HCN. Analysis for ^C ₃ ^H 36 ^N 2 2 * ¹ ° 12 ^'5 ° ^H 2 Calculated: C 57.57, H 5.89, N 4.20% Found: C 57.35, H 5.94, N 3, 82%.

The compounds according to the invention can be used in mammals against cancers. Their efficacy has been verified by in vivo and in vitro experiments. In one in vivo experiment conducted by the Cancer Chemotherapy Reporte, National Cancer Institute, 3, No. 2, Part 3, September 1972, a certain amount of P-388 lymphocytic leukemia was intraperitoneally inoculated into healthy mice.

After inoculation, mice were treated on days 5, 9 and 13 with varying amounts of the compounds produced by the method of the invention. Controls were untreated mice, other mice were treated for comparison with daunorubicin or doxorubicin: 3 ^from deamino-3 '- (4'-яогfolinyl) daunorubicin or 3-deamino-3 ^* - (4 "-morpholinyl) -13-dihydrodaunorubicin by US No. 4,301,277 or 3'-deamino-3'-4'-methoxy-1-piperidinyl) daunorubicin or its 13-dihydro equivalent to U.S. Patent No. 4,314,054.

The average survival time in the various treated mice was determined and compared to the 8 untreated control animals. The following Table A shows the data obtained as% T / C, which is the survival time of the treated mice divided by the survival time of the control mice x 100. As shown in Table A, various doses of the individual compounds were monitored to achieve the greatest improvement.

Table A ·

Efficacy against Leukeeia p-388 in higher L-1210 cells ·

compound NSC δ. ^Χ Survival time X T / C Optional dose (44d, 5, 9, 13) (ag / kg) Inhibition of ^ED 50 DNA synthesis / UM RNA 3 ^x -deanino-3 ^/ - (3'-cyano-4 “** - -юогfolinyl) daunorubicin 332,304 197 0.4 0.12 0,002 3 ^z -deanino-3 * - (3'-cyano-4-morpholinyl) -3-dihydrodaunorubin 332,305 143 ’ 0.1 0.019 0,002 3 * -deaDino-3 * - (3 * -cyan-4‘- -raor folinyl) doxorubic in 357.704 187 0,075 0.003 0.0005 3 * -Deamino-3 (3-cyano-4-morpholinyl) 13-dihydrodoxorubicin 360,291 150 ' 0.2 0,021 0,0030 For comparison: daunorubic in 82,151 130 8 0.66 0.33 doxorub ic in 123,127 130 8 1.5 0,5S! 1

^X NSC No. «National Sevice Center registration number of the US National Cancer Institute

g 3 ^x deamino-3 '- (4 • -morfoliny1) - /

-daunorubicln • 327,451 165 0.2 0.76 0.10 · _; 3 ^x -deamino-3 * - (4'-morpho- t i. Lnyl) -1,3-dihydrodauno- rubicin 327,450 132 0.2. 2.2 0.67

CS 268512 В2

The results show that the compounds produced by the process of the invention have good to very good antitumor activity in vivo at low optimal doses, compound No. NSC 357 704 has an optimal dose of about 1/150 of the optimal dose of the parent compounds. Other compounds produced by the process of the invention have an optimum dose far lower than daunorubicin and doxorubicin, which is promising in the search for anti-cancer agents with reduced cardiac muscle toxicity.

In vitro assays using 3 ^of -deamino-3 '- (3-cyano-4'-morpholinyl) daunorubulin and 3 ^of -deamino-3 ^of - (3'-cyano-4'-morpholinyl) -13 These materials have been investigated as inhibitors of DNA and RNA synthesis in L-1210 leukemia, as described by G. Tong, W.W. Lee, D.W. Black, and DW Henry. Medicinal chem., 19, 395 (1976). The compounds were effective at doses up to 600 times lower than the required doses of daunorubicin, doxorubicin or their previously known analogs. It was also observed that it inhibited RNA synthesis more than DNA ED 50 for a DNA / RNA ratio of 10 to 11. Oak was reported in ST Crooke et al., Mol. pharmacol., 14, 290 (1978), such a ratio demonstrates α class II anthracycline of good therapeutic properties, inhibition data are also shown in Table A.

The data in Table A show the increased antitumor activity of derivatives with a morpholine structure and a cyancmorpholine structure, which is obviously typical of this class of substances.

Additional in vivo tests of mice against P-388 and L-1210 leukemia and B-16 melanoma were performed according to cancer chemotherapy Reporte 1972, above. B.

Table В

Antitumor efficacy of wells at the optimal dose of% T / C (eg / kg)

NSC compound δ. daunorubicin 82151 * doxorubicin 123127 * 327451 (3) * 327450 * 332304 332305. ip L1210 d 1> ip 168 (4) ‘ q3d 1,5,9 / ip qd 1-9, Ip 173 mm (1) ip P388 d. 1, ip 252 (7.5) 155 (0.25) 175 (0.25) d. 1, ip 185 (9) 152 (0.5) ' 157 (0.5) q4d 1,5,9, ip 257 (3) q4d 1,5,9, AFTER qd 1-9 > 285 (1) ip B16d d. 1, ip > 197 (19) q4d 5,9,13, ip 129,146 (2) ip B16 d. 1, ip 195 mm (4) 292 (3) 135 (0.25) 121 (0.25) 138 (0.15) 131 (0.0375)

compound

NSC no. 354647 (2) * 355278 (3) 357704 360291 ip L1210 d. 1, ip 152 (0.05) 164 (0.025) 152 (0.025) q4d, 1.5.9, ip > 164 (0,0125) 128 (0.05) Qd 1-9 > 141 (0,026) 166 (0.0063) / 148 (0.025) ip Ρ3Θ8 d. 1, ip 182 (0.05) > 193 (1) 262 (0.0125) 183 (0.05) dl, i v > 143 (0.062) 166 (2.5) 152 (0.05) q4d 1.5.9; 224 (0.05) q4d 1,5,9, po 175 (0.05) > 158 (0.1) qd 1-9 209 (0.0125) • ^ 360 (0.0063) 267 (0.025) ip B16d d. 1, ip > 138 (0,0064) 149 (0.025) q4d 5,9,13, ip 123 (0.0125) 121 (0.025) 1 lp 816 d. 1, ip

Explanatory notes to Table В:

^x known compounds for comparison (1) ^of 3 -dea ^"of amino-3 - (4'-morpholinyl) -13-dihydrodaunorubicin (point 4 of U.S. Patent No. 4 031 277) (2) 3--deaaino ^{of from} 3 - (4 (3 ') 3'-deamino-3 ^from - (4'-morpholinyl) -13-dihydrodoxorubicin (item 5 of U.S. Patent 4,301,277).

The compounds produced by the method of the invention, including their salts, can be administered by any route, for example, orally and parenterally, i.e., intravenously, intraperitoneally, subcutaneously, or intramuscularly. Parenteral administration, particularly intravenous administration, is particularly preferred. Dosages and amounts as described above can be used to improve leukemia or other type of cancer against which they are effective. For example, in the treatment of small experimental animals, doses of 0.0010 to 25 mg / kg per day may be used to improve leukemia. The upper limit of dose is the dose which may already produce toxic side effects and is determined by trial and error in test animals. Typically, the dose of the compounds produced by the method of the invention is about 1/20 to 1/200 of the parent compounds. While a single dose may be administered every 2 to 7 days, shorter intervals between doses may be used.

The compounds of the present invention and their salts can be formulated into pharmaceutical compositions, particularly in unit doses. The compounds may be administered as such, more commonly administered together with a pharmaceutical carrier that dilutes the compound with ease of administration. It will be appreciated that this carrier, like the resulting formulation, must be sterile and non-toxic. .

! · '

Л

For oral administration, the carrier can be used as a diluent, solid, semi-solid or liquid and is the environment in which the active ingredient is used for pharmacological experiments. For parenteral administration, the active ingredient is dissolved or suspended in a suitable injection liquid in a manner known per se.

Conventional methods of treating water-soluble pharmaceuticals for salts and water-insoluble substances for free bases can be used in the manufacture of the above dosage forms. Injectable compositions may be obtained, for example, as follows.

Composition A

A sterile suspension in an aqueous medium catalyzes from the following mixture:

component compound of example 1, 2, 3, 4, 5, 6, 9 or 10 in suspendable powder sodium citrate sodium carboxymethylcellulose (low viscosity) methylparahydroxybenzoate propylperahydroxybenzoate

The above mixture is made up with water which is suitable for injection by injection

5.7

2,0

1/5

0.2 · suitable for injection to a volume of 1.0 ml.

Composition A ^of

Sterile suapency in an aqueous environment suitable for injection from the following mixture.

Component cg can be obtained

3 ^of -deamino-3 ^of - (3'-cyan-4'-morpholine l) doxorubicin of Example 4

0.5 sodium citrate

5.7 Carboxymethylcellulose sodium (low viscosity)

2,0 methyl parahydroxybenzoate

1,5 propylparahydroxybenzoate

0.2

The above mixture is supplemented with water suitable for injection to a volume of 1.0.

Means В

The capsules can also be obtained from the following mixture:

s bed mg

3 ^of -deamino-3 ^of - (3'-cyano-4'-morpholinyl) doxorublinIn of example 4 (C) lactose

199

The compound of Example 4 (C) and lactose are passed through a sieve and the powder mixture is mixed well, then filled into hard gelatin capsules of suitable size by placing 200 g of the powdered material obtained in each capsule.

Claims (1)

SUBJECT YOU FINISH 1. A process for the preparation of novel morpholino derivatives of the formula I of daunorublcin and doxorubicin of general formula (I). CS 268512 82 where R. is -CO-CH ₃ , -CHOH-CH ₃ , -CO-CH ₂ OH and -CHOH-CH ₂ OH, characterized in that a compound of the formula MÍ kde R represents a group -CO-CH ₃ , -CO-CH ₂ OH, CHOH-CH ₃ or -CHOH-CH ₂ OH in a mixture of water and a polar organic medium with 2,2'-oxydiacetaldehyde of the formula prepared in situ by cleavage of 1,4 The reaction is carried out in the presence of an alkali metal cyanoborohydride, for example sodium or potassium cyanoborohydride, and the crude reaction product obtained after closing the morpholine ring is separated into neutral and alkaline fractions and the neutral fractions are chromatographically purified, thereby purifying the neutral fractions. the pure product of formula (I) is obtained.