HU193879B - Process for preparing morpholinyl-daunorubicin and morpholinyl-doxurubicin derivatives and pharmaceuticals comprising such compounds as active substance - Google Patents

Abstract

The present invention relates to a compound of formula (I): wherein R is CO-CH3 or CHOH-CII3 in the case of the derivative of danuborubicin or CO-CH2OH or CHOH-CH2OH in doxorubicin derivatives; the preparation. The process of the present invention is carried out by treating a compound of formula (III) wherein R is -CO-CH 3 or -CO-CH 2 OH in water and a polar organic solvent in a mixture of 18 molar equivalents of 2,2'-oxoacetaldehyde of formula (1). are reacted in situ from 1,4-anhydro-erythritol at room temperature, cleaved with sodium periodate, in the presence of two molar equivalents of alkali metal cyanoborohydride to the starting material, and the crude product which is basic morpholine derivatives and 13-dihydro analogues of the starting material and the neutral neutral 3 "cyano-morpholine analogues are separated into basic and neutral fractions. The neutral fractions are chromatographed on a silica gel column eluting with a separate 9: 1 dichloromethane / methanol mixture to give the first eluate. the desired 3-cyano-morphine of formula (I) an olino derivative wherein R is -CO-CH 3 or -CO-CH 2 OH and the following eluant is the corresponding 3 "-cyano-13-dihydro derivative of formula I wherein R is -CHOH-CH 3 or CHOH -CH2OH- and the resulting product is purified if desired. -1-

Description

The present invention relates to morpholinyl-daunorubicin and morpholinyl-doxorubicin derivatives having antitumor activity and their analogs.

The invention described herein was developed by the U.S. Department of Health and Human Services National Institutes of Cancer Grant Nos. CA25711 and CA32250.

The present invention relates to the field of anthracycline chemistry, more particularly to the preparation of anthracycline doxorubicin and daunorubicin analogs, which are antitumor agents.

Doxorubicin (adriamycin) is disclosed in U.S. Patent No. 3,590,028. It is described in F. Arcamone et al., U.S. Pat. This compound is probably the most useful new anticancer drug today. Together with daunorubicin, it is the most important drug for the treatment of a variety of solid tumors and leukemia. Unfortunately, however, many patients with tumors do not respond to this drug and do not respond substantially to patients with certain serious tumor types such as spinal cancer and melanoma. In addition, in some patients, chronic treatment can cause irreversible cardiac damage, which can be fatal if continued. Thus, there is a great need for the preparation of analogues which produce a better response in a broad spectrum or show reduced cardiotoxicity. There is a great demand for more potent and less toxic agents, and it is therefore an object of the present invention to provide such agents. Based on the screening results, two lipophilic derivatives (AD32 and N, N-dibenzyl-daunorubicin) proved to be the most effective novel analogs in a 3-dose treatment line (q4d 5, 9, 13) in a widely used anti-mouse leukemia test (q4d 5, 9, 13). they were not reactive with DNA in vitro, although DNA is a primary biological target in the anthracycline series. Most N-alkyl derivatives were effective in the anti-tumor screening assay against murine leukemia P-388 but not significantly different from doxorubicin or daunorubicin. Some of these derivatives have been shown to be ineffective.

The literature on doxorubicin and anthracycline analogues can be found in the article "Adriamycin" by David W. Henry, ÁCS Symposium Series, No.30, Cancer Chemotherapy, American Chemical Society, 15-57. (1976), and Doxorubicin, by Federico Arcamone, Academic Press, 1981. AD32 is incorporated by reference 4,035,566. U.S. Pat.

No. 4,301,277. U.S. Pat. No. 3,600,123 discloses 3'-desamino-3 '- (4-morpholinyl) -daunorubicin by Edward M. Acton and Carol W. Mosher, an effective dose of eve2 as a quarter of the effective dose of doxorubicin, but essentially azo. gave a T / C value (166%) of 160% P388. The compound and its preparation and properties are described in Enhanced Antitumor Properties of 3 '- (4-Morpholinyl) and 3' - (4-Methoxy-1-piperidinyl) Derivatives of 3'-Deamino-daunorubicin J. Med. Chem. , 25, 18-24. (1982), Carol W. Mosher, Heten Y. Wu, Allan N. Fujiwara, and Edward M. Action.

A general reductive alkylation process for the preparation of novel semisynthetic anthracycline derivatives has been described in Adriamycin Analogs. 3. Synthesis of N-Alkylated Anthracyclines with Enhanced Efficacy and Reduced Cardiotoxycity J. Medicinal Chem., 22, 912-918. page (1979),

G. L. Tong, H.Y. Wu, T.H. Smith and D.W. Henry.

J. Med. Chem. 25, 18-24. pages 1982, by Carol W. Mosher et al., inventors of the present application, were directed to the preparation of daunorubicin analogs by incorporating the 3'-nitrogen of the sugar moiety into a new ring (morpholinyl ring). diacetaldehyde in the presence of NaBH ₃ CN by reductive N, N-alkylation. According to this article, 3'-desamino-3'- (4-morpholinyl) -daunorubicin and its 13-dihydro-derivative have been isolated, which is partially due to the reduction of the 13-keto group of the starting material by the presence of sodium cyanoborohydride. However, it is also apparent from the article that significant amounts of by-product were formed during the reaction which, in the absence of basicity, were not utilized as an acid insoluble product.

The present invention provides novel daunorubicin and doxorubicin derivatives of formula (I).

In the formula I, R is -CO-CH ₃ or -CHOH-CH _{3 for} daunorubicin derivatives or

-CO-CH ₂ OH or -CHOH-CH ₂ OH for doxorubicin derivatives.

Compounds of formula (I) are prepared by reacting a known daunorubicin, doxorubicin of formula (III), and its analogs wherein R is COCH ₃ , -CO-CH ₂ OH-water, and a polar organic solvent (1). and the appropriate precursor thereof, and the reaction is carried out in the presence of a cyanoborohydride salt, such as an alkali metal cyanoborohydride, and the desired compounds are isolated and purified in known manner. These compounds are related to the known anticancer agents daunorubicin and doxorubicin (adriamycin, from which they can be used to synthesize and transform useful anticancer agents. The new compounds combine a high degree of antitumor activity and a low dose of the compounds). 2193876 because of their high potency and the fact that low doses reduce side effects, such as cardiotoxicity, compared to the above-mentioned substances. They included.

Thus, the present invention provides cyanomorpholinyl derivatives of daunorubicin and doxorubicin. These compounds are listed in Table I below.

Compounds of formula (I) prepared according to the invention

R Name of the compound j

-CO-CH3 3-desamino-3- [3 '] -cyano-4-morpholinyl / -dunorubicin

-CHOH-CH3 3 '-Deamino-3 * - [3-cyano-4-morpholinyl] -13-dihydro-daunorubicin

-CO-CH2OH 3'-desamino-3 '- / 3-cyano-4-morpholinyl / ~

doxorubicin

-CHOH-CH2OH 3 '-Deamino-3' - / 3'-cyano-4-morpholinyl / -13-dihydrodoxorubicin

The first of the four materials is the most preferred.

Preparation of some preferred compounds

These compounds can be prepared as follows at ³ °:

First, the commercially available daunorubicin or doxorubicin, in the form of its acid addition salt, is reacted with 2,2'-oxydiacetaldehyde (1) under reductive alkylation conditions. This alkylation results in a product mixture containing four main components. These components for daunorubicin are as follows: ₄ q

3'-desamin'o-3 -'- (4-morpholinyl) -daunorubicin 3'-desamino-3 '- (4-morpholinyl) -13-dihydro-daunorubicin,

3'-desamino-3'- (3-cyano-4-morpholinyl) daunorubicin and

3'-desamino-3'- (3-cyano-4-morpholinyl) -13-dihydro-daunorubicin.

In the case of doxorubicin, the product contains the following components:

3'-desamino-3'- (4-morpholinyl) -doxorubicin, ₅₀ 3'-desamino-3'- (4-morpholinyl) -13-dihydrodoxorubicin,

3'-desamino-3'- (3-cyano-4-morpholinyl) doxorubicin;

3'-desamino-3 '- (3-cyano-4-morpholinyl) -13-gg-dihydro-doxorubicn.

2,2'-Oxydiacetaldehyde can be prepared from 2,2'-oxydiacetaldehyde bis (diethylacetal) by acid hydrolysis Eleid et al., U.S. Patent No. 655,436. or by cleavage of 1,4-anhydroerythritol by Barry et al., Carbohydrate Research, 7,

299 (1968) and Carbohydrate Research, Greenberg et al., 35, 195 (1974).

Reductive alkylation may be carried out in an aqueous polar organic solvent mixture such as an excess of water and acetonitrile, typically at about pH 7, in the presence of a reducing agent such as an alkali metal cyanoborohydride such as sodium or potassium cyanoborohydride. This is a relatively mild reaction that can usually be carried out at room temperature for 1 hour or less. Reductive alkylation is exemplified and described in U.S. Patent No. 4,301,277. U.S. Pat. Chem., 25, 18-24. (1982).

The reaction product mixture can be worked up by any method which results in isolation and isolation. It is effective if the reaction product is extracted with acid so that acid-extractable non-cyano-substituted materials can be isolated from acid-insoluble cyano-substituted materials. The resulting pairs of materials can then be separated by different chromatographic methods, such as preparative layer chromatography, column chromatography or preparative high pressure liquid chromatography.

Further details of the invention are illustrated by the following examples.

Example 1

3'-Desamino-3'- (3-cyano-4-morpholinyl) -daunorubicin and 3'-desamino-3 '- (3-cyano)

Preparation, Isolation and Identification of 4-Morpholinyl) -13-dihydro-daunorubicin A solution of 1,4-anhydroerythritol (31.5 g, 0.30 mol) in 400 mL of water was stirred and cooled in small portions. NaIO ₄ (0 g, 0.25 mol) was added. Stirring was continued at room temperature for another hour, with small amounts

-3193879

NAIO ₃ is released. The mixture was allowed to stand and the pH was adjusted to 7.3 by the addition of a few drops of sodium bicarbonate solution and the dissolution was completed by the addition of 500 ml of water. The 2,2'-oxydiacetaldehyde thus formed is not isolated.

To the dialdehyde solution was added a solution of 9.2 g (16.3 mmol) of daunorubicin · HCl in 50 mL of a 1: 1 mixture of CH ₃ CN and water and stirred at room temperature. The mixture becomes cloudy and a small solid precipitates after 45 min. Adding a further 75 mL of methyl cyanide partially solubilizes the solution with 2.02 g (32.6 mmol) of sodium cyanoborohydride in 6 mL of water. After stirring for 1 hour, dilute with water and extract with chloroform (5 x 200 ml). 0.91 g of solid which was insoluble in each phase was removed by filtration.

After evaporation of the chloroform, the crude reaction product was 3'-desamino-3'- (4-morpholinyl) dau-norubicin, 3'-desamino-3'- (4-morpholinyl) -13-dihydro-daunorubicin, 3 ^z -desamino-3 Contains' - (3-cyano-4-morpholinyl) -daunorubicin and 3'-desamino-3'- (3-cyano-4-morpholine-1) -13-dihydrodunorubicin. The solid crude product (5.41 g) was dissolved in chloroform / methanol (9: 1, 500 mL). The solution was washed with 3 x 100 mL of 0.01 N hydrochloric acid, 1 x 100 mL of water and 1 x 100 mL of dilute sodium bicarbonate. The organic phase is dried, evaporated to dryness and the residue is dried under vacuum (0.1 mm Hg) at room temperature. 5.10 g of a glassy residue are obtained.

The residue (5.09 g) was dissolved in dichloromethane: methanol (4: 1, 50 mL). The solution was stirred until 30 mL of methyl cyanide was added dropwise. The cloudy solution was evaporated to dryness to give a semi-solid residue which was triturated with 200 ml of methyl cyanide in the dark. The insoluble solid was collected and triturated a second time with 100 mL of methyl cyanide. The liquid phase obtained after two rubbing operations contains the desired material. After evaporation, 2.23 g of semi-solid residue were 3'-desamino-3'- (3-cyano-4-morpholinyl) -daunorubicin and 3'-desamino-3'- (3-cyano-4-morpholinyl) -13-dihydro. a crude mixture of daunorubicin is obtained.

The semi-solid residue was then dissolved in 5 mL dichloromethane and applied to a column of 3.1 cm x 59 cm 0.074 mm to 0.044 mm Mallinckrodt Silic AR CC-7 silica gel washed with dichloromethane. The column was eluted with 500 mL dichloromethane followed by dichloromethane: methanol (1500 mL 99: 1, 1000 mL 98: 2, 1500 mL 97: 3 and 500 mL 90:10). Fractions (10 ml) were collected (monitored by TLC) from the initial eluate (2550 ml), and the fraction (190 ml) was evaporated to give 0.48 g of product. The principal component was determined by comparative high performance liquid chromatography (TLC) and thin layer chromatography ( ₇₎ and indicated that the product was pure 3'-desamino-3 '- (3-cyano-4-morpholinyl) -daunorubicin.

Further, 0.35 g of the above sample was further purified in the dark for the first time on five 2 mm x 20 x 20 cm silica gel plates, using dichloromethane: methanol (29: 1) twice as eluent. The middle band containing 65% of the weight of the sample is cut out and eluted, filtered and evaporated to dryness, and the same purified materials are recovered from the chromatographic side adjacent zones and 0.18 g is further purified by a 1.1 cm x 27 cm on a 0.074-0.037 mm silica gel column. The column was eluted with dichloromethane / ethyl acetate (30 mL, 80:20, 30 mL, 60:40, and 30 mL, 80:20) and finally eluted with 175 mL of ethyl acetate. 16 ml of the initial eluate was collected in 2 ml fractions and monitored by TLC, followed by collection of 88 ml fractions and evaporation. 0.15 g of product is obtained.

Elemental analysis showed 3'-desamino-3'- (3-cyano-4-morpholinyl) -daunorubicin as the pure product. The same structure is confirmed by 360 MHz, NMR, IR and mass spectroscopy.

The formation of the product in diastereomeric form is indicated by HPLC and 360 MHz NMR. HPLC analysis on a Waters Radial Shake C-18 column with 0.5 M pH 4 citrate buffer methyl cyanide 60:40 at 2 ml / min shows two close peaks at 9.6 minutes and 10.2 minutes at 53:44. The 360 MHz NMR spectrum of this material showed two resonances at 6-OH, 11-OH, 1-H, 2-H, 3-H, 1'-H, 7-H, 9-OH, 10A-H, -H ₃ and 6-H ₃ protons.

NMR 360MHz CDC1 ₃ he 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, 7-H), 4.47 * , 4.34 * (2s, 9-OH), 4.08 (s, OCH ₃ ), 3.97-4.07 (m, 2BH), 3.92 (t,

J = 12Hz 3-H), 3.74 (m, 2A-H), 6B-H), 3.68 (bs, 4Ή), 3.58 (t, J = 12Hz, 6A-H), 3, 20 (d, J = 19Hz, 10B-H), 2.91, 2.90 (2d,

J = 19Hz, 10A-H) 2.75-2.95 (m, 3'-H), 2.68 (m, _5-H2), 2.43, 2.42 (2s, 14-H ₃ ), 2.35 (m, 8B-).

After eluting the 3'-desamino-3'- (3-cyano-4-morpholinyl) -daunorubicin, the Mallinckrodt Silic ARCC-7 silica gel column was further eluted with 90:10 by volume CHCl3: methanol. After collecting 3460 ml of eluent, a 430 ml fraction consisting essentially of a single substance was collected and evaporated, containing 12.5% of the initially fed material. The compound is not pure 3'-desamino-3'- (3-cyano-4-morpholinyl) -13-dihydro-daunorubicin, which is further purified by the same method as the non-hydrogenated analog and substantially pure 3'-4193879

-deamino-3'- (3-cyano-4-morpholinyl) -13-dihydro-daunorubicin is obtained.

NMR 360MHz CDC1 _3: similar to that of 3'-deamino-3 '- (3-cyano-4-morpholinyl) -daunorubi'cinéhez, except that 14-H ₃ 2.43 resonances, δ 2.46 2s) upwards are offset (1.32, 1.30, 2d, J = 64 Hz). Mass spectrum (such as trimethylsilyl derivatives (TMS), m / e 957 [M / TMS / _5- HCl] 885 [M / TMS / _4- HCl] TLC SiGel CHCl ₃ (19: 1) R f = 0, 28th

Example 2

Preparation and Isolation of 3'-Desamino-3'- (3-cyano-4-morpholinyl) doxorubicin and 3'-desamino-3 '- (3-cyano-4-morpholinyl) -13-dihydro-doxorubicin

Sodium metaperiodate (6.42 g, 30.0 mmol) was added to a solution of 6.25 g (60.0 mmol) of 1,4-anhydro-erythritol in 75 mL of water-cooled water bath at 15-20 ° C. The resulting clear solution was stirred at room temperature for 17 hours. The solution was adjusted to pH 4.0 with sodium bicarbonate and diluted with methyl cyanide (75 mL) with stirring. Precipitation occurs. The mixture containing 2,2'-oxydiacetaldehyde was stirred and 0.126 g (2 mmol) of sodium cyanoborohydride in 5 ml of methyl cyanide / water 1: 1 was added. To the mixture was added 1.1xg (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 and extracted three times with 50 mL portions of chloroform. The crude extract was 3'-desamino-3'- (4-morpholinyl) -doxorubicin, 3'-desamino-3'- (4-morpholinyl) -13-dihydro-doxorubicin, 3'-deamino. Contains 3 '- (3-cyano-4-morpholinyl) doxorubicin and 3'-desamino-3'- (3-cyano-4'-morpholinyl) 13-dihydro-doxorubicin. The combined extracts were extracted with 5 x 25 mL of 0.1 N acetic acid and water, washed with dilute sodium bicarbonate and saturated aqueous sodium chloride. The acidic aqueous phase was separated. The chloroform layer was filtered through sodium sulfate and evaporated. The residue obtained is dissolved in 25 ml of chloroform and the solvent is again evaporated in vacuo at room temperature. 0.518 g (40%) of a dark red glass foam were obtained.

0.424 g of this glass foam was dissolved in 1.5 ml of dichloromethane and applied to a 0.074-0.037 mm Bio-Sil A silica gel column washed with 1.5 x 35.5 cm @ 3 of dichloromethane. The column was eluted with 50 mL dichloromethane followed by dichloromethane / methanol 150 mL 99: 1, 150 mL 98: 2, 300 mL 97: 3, 100 mL 95: 5 and 300 mL 90:10. After collecting the initial eluate (445 ml), the 80 ml fraction was evaporated to give 0.217 g of product. The material is combined with 0.039 g of the purified product obtained previously and the mixture is dissolved in 2 ml of dichloromethane, diluted with 10 ml of methanol and evaporated to dryness. The residue was triturated with 5 mL of methanol 0.218 g

3'-Desamino-3'- (3-cyano-4-morpholinyl) doxorubicin is obtained.

The product was diastereoisomerated by HPLC and 400 MHz NMR. HPLC analysis on a Waters Radial-Pak C-18 column with 0.05 M pH 4 citrate buffer methyl cyanide (65:35) at 2 ml / min shows 2 near peaks at 14.4 minutes and 15, 7 minutes at 58:39. The 400 MHz spectrum exhibits two resonances at 1-H, 2-H, 3-H, 1'-H

7-H, 14-H ₂ , 9-OH, OCH ₃ , 10A-H and 6'-H ₃ .

400 MHz NMR CDC1 ₃ δ 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, 1'-H), 5.34, 5.30 (2m, 7-H), 4.79,

4.78 (2s, 14-H2), 4.54, 4.42 (2s, 9-OH), 4.11,

4.10 (2s, OCH3, 4.05 (m, 5'-H). 3.97 (m,

2B-H, 3-H, 6B-H), 3.71 (m, 4'-H, 6-AH), 3.58 (t, 2A-H), 3.30 (d, J = 19 Hz) , 10B-H), 3.07, 3.06 (2d, 10A-H), 3.03 (m, 3'-H), 2.69 (m, 5-H, 2.38 (m, 8B) -H), 2.22 (m, 8A-H), 1.84 (m, 2'-HJ, 1.61 (s, H ₂ O), 1.40, 1.39 (2d, J = 6). , 5 Hz, 6'-HJ.

UV-Vis _(CH3 OH) max 234 nm), 40.100), 252 (27.700), 289 (9.420), 478 (13,000) 495 (13,000) 495 (12,900), 530 (7.190). Mass spectrum (trimethylsilyl) (TMS) as derivatives), m / e 899 (TMS) _4- HCN. Calcd for C 3 H ₃₄ N ₂ O ₂ 2H ₂ O ₁₂ Calculated:% C 56.97% H 5.68% N 4.15% Found: C 57.07% H 5.37% N 4, 17th

Further elution of the above column yielded 0.041 g of 3'-desamino-3'- (3-cyano-4-morpholinyl) -13-dihydro-doxorubicin.

UV-Vis (CH ₃ OH) λ max 234 nm (ε 37,400), 252 (28,000), 289 (9,390), 475 (12,700), 496 (12,800), 530 (7,410).

Mass spectrum (as trimethylsilyl (TMS) derivatives), m / e 985 M (TMS) ₅ -CH ₃ , 973 M (TMS) ₅ -HCN

Analysis for C ₃₂ H ₃₆ N ₂ O | Calcd O ₂ -1.5 H ₂ O: 57.57% C 5.89% H 4.20% N Found 57.35% C 5.94% H 3.82% N.

The compounds of the present invention may be used as an antitumor agent in mammals. This effect was demonstrated by in vivo and in vitro assays. In one of the in vivo tests performed by Cancer Chemotherapy Reports, National Cancer Institute 3, No. 2,

Part 3, September 1972, healthy mice were inoculated intraperitoneally with Lymphocytic leukemia P-388 ascetic fluid. The vaccinated mice are then exposed to the 5th,

On days 9 and 13, various amounts of the compound of the invention were treated. for comparison, no other mice were treated and additional mice were treated with daunorubicin, doxorubicin, or mice described in U.S. Patent No. 4,301,277. 3'-desamino-3 '- (4-morpholinyl) -daunorubicin or 3'-desamino-3'- (4-morpholinyl) -13-dihydro-dunorubicin or U.S. Patent No. 4,314,054. United States of America

3'-desamino-3 '- (4-methoxy-1-piperidinyl) -daunorubicin or its 13-dihydro equivalent described in U.S. Patent No. 5,193,879.

The average survival time of the mice treated in different ways was determined and compared with the survival time of mice inoculated with the leukemic ascites fluid, which were not treated with the test compounds. Table A below shows the data obtained. Data are expressed as T / C%, which is the survival time of treated mice divided by the survival time of control animals multiplied by 100. In Table A, dose levels of the various substances that contributed most to the increase in survival time are also given.

The table

Leukemia P-388 Leukemia Activity against L-1210 cells in mouse

Compound

Survival time

Optimal dose _{/ q} 4d, 5,9,13 /

Synthesis inhibition ED salt juM

NSC No *% T / C mg / kg DNA

RNA,?

-desammo-3- (3-cyano-4-morpholinyl) -dunoro-

bicin 3 ¹ -deamino-3 ⁷ - [(3-cyano-4-morpholinyl) -13-dihydro] 332 304 197 0.4 0,012 0,002 ro-daunorubicin 3'-desamino-3 ^; - - (3-cyano-4-morpholinyl) -doxoro- 332.305- 143 0.1 0.019 0,002 bicin 3 '-deamino-3 ^; - - (3-cyano-4-morpholinyl) -13-dihydro- 357 704 187 0,075 0,003 0.0005 ro-doxorubicin For comparison: 360 291 150 0.2 0,021 0.0030 daunorubicin 82 151 130 8 0.66 0.33 doxorubicin 3 ⁷ -deamino-3 '- (4-morpholinyl) - 123 127 160 8 1.5 0.58 -daunorubicin 3 ^? -deamino-3 '- (4-morpholinyl) -13-dihydrodauno- 327 451 166 0.2 0.76 0.10 doxorubicin 327 450 132 0.2 2.2 0.67

* NSC No, = US National Service Center National Cancer Institute Registration Number

The results show that the compounds of the present invention exhibit good, even θθ, in some cases excellent in vivo antitumor activity at optimally low doses. NSC 357704. compound ca. It was 1/150 times more potent than the parent compound. Other substances of the present invention are also more potent at levels much lower than daunorubicin and doxorubicin. Therefore, good antitumor agents with significantly lower cardiotoxicity are obtained.

3'-desamino-3'- (3-cyano-4-morpholinyl) -daunorubicin and 3'-desamino-3 '- (3-cyano-4-morpholinyl) -13-dihydrodaunorubicin in

-6193879's in vitro assays also show enhanced activity in this class of compounds. When these substances were tested for inhibition of DNA and RNA synthesis in L-1210 cells by G. Tong,

W. W. Lee, D.R. Black and D.W. Henry Mask, as described in J. Medicinal Chem., 19, 395 (1976), was found to be very effective at doses 600 times lower than those used for daunorubicin, doxorubicin or other prior analogues. They showed a greater inhibitory effect on RNA synthesis than on DNA synthesis (ED 50 DNA / / RNA = 10-11). S.T.Crooke and others on the Mole. Pharmacol., 14, 290 (1978). better anthracycline classB. table y properties. These data are taken from Table A.

The data in Table A, indicating increased anti-tumor activity on the morpholine structure and further enhanced activity on the cyanomorpholino structure, characterize the effect of this group of compounds.

Further tests were performed to confirm the biological activity of the compounds. These were in vivo tests in mice against P-388 and L-1210 leukemia and B-16 melanoma, performed according to the method described above in Cancer Chemotherapy Reports, 1972. At various doses i.p., i.v. and tests were performed using the oral route. The results are shown in Table B.

Anti-tumor effect in mouse at optimal dose 7, T / C / mg / kg /

Compound dauno- doxo-

NSC No. rubicin rubicin 82151 * 123127 * 327451/3 / ^x 327450/1 / * 332304 332305 354646/2 / * 357704 360291 JUICE L1210 d 1 , op 168/5 / 152 / 0.05 / 164 / 0.025 / 152 / 0.025 / q'd 1,5,9, ip > 164 / 0.0125 /> 128 / 0,05 / qd 1-9, ip 173/1 / > 141 / 0.025 / 166 / 0.0063 /> 148 / 0.025 / ip P388 d 1 , iP 252 / 7.5 / 155 / 0.25 / 175 / 0.25 / 182 / 0.05 / 262 / 0.0125 / 183 / 0.05 / d 1 , iv 185/10 / 152 / 0.5 / 157 / 0.5 / > 143 / 0.062 / 152 / 0.05 / Cp Hq 1.5.9 ip 257/4 / 224 / 0.05 / q a 1.5,9, after 175 / 0.05 / > 185 / 0.1 / qd 1-9, ip > 285/2 / 209 / 0.0125 /> 360 / 0.0063 / 267 / 0.025 /

ip B16d d 1, ip> 197/20 / q ^ dS ^ LVp 129,

146/3 / ip B16

38 / 0.0063 / 149 / 0.025 / 123 / 0.0125 / 121 / 0.025 /

195/4 / ^x Known compounds / 1/3 * -deamino-3/2/3 ^? -deamino-3/3/3 -deamino-3

292/4/135 / 0.25 / 121 / 0.25 / 138 / 0.25 / 131 / 0.375 / for comparison purposes, "/ 4" -morpholinyl / -13-dihydrodunorubicin / See claim 4, No. 4,301,277 / / see Claim 6 of No. 4,301,277 /

- / 4 "-morpholinyl / doxorubicin

- (4-morpholinyl) -13-dihydrodoxorubicin / See claim 4, No. 4,301,277 /

US Patent Specification

US Patent Specification US Patent Specification

The compounds of the present invention, including their salts, may be administered by any route, including oral, parenteral (intravenous, intraparitoneal, subcutaneous and intramuscular) routes. The top 50 have traditionally been the parenteral route of administration, especially intravenous administration. The dosage range and amount are sufficient to ameliorate leukemia or other cancers against which the compounds are active. For example, in the treatment of test animals, a dose of 0.0010 mg / kg to 25 mg / kg was sufficient to ameliorate the condition of the leukemic patient daily. The upper dose limit is determined by toxic side effects. This can be determined in animal θθ experiments. Generally, it is 1/20 to 1 200 times lower than the parent compounds. There may be an interval of 2 to 7 days between two doses, but shorter intervals, such as one day or less, may be employed.

For ease of administration, the compounds of the present invention, or salts thereof, are prepared in the form of pharmaceutical compositions, particularly unit dosage forms. Although the compounds may be administered alone, they are usually administered in a pharmaceutically acceptable carrier, which dilutes the compound and facilitates treatment. The term "pharmaceutically acceptable" means that the carrier and the resulting composition are sterile and non-toxic.

For oral administration, the carrier or diluent may be solid, semi-solid or liquid and may serve as a vehicle or excipient for the active ingredient. For parenteral administration, the material is dissolved or suspended in a suitable injectable liquid as is known in the art.

For the preparation of dosage forms, the known technology for the preparation of water-soluble drugs may be used in the case of salts and water

-7193879 For insoluble agents, the technology used is the free base. For example, injectable materials are prepared as follows.

The preparation:

Made in water, injectable sterile suspension mg The compound of Examples 1, 2 can be suspended as a powder 3 IC Sodium citrate Sodium carboxymethyl 5.7 -cellulose (low viscosity grade) 2.0 15 Methyl para-hydroxybenzoate 1.5 Propyl p-hydroxybenzoate Water for injections up to 1 ml 0.2 Preparation A: Injectable in aqueous medium sterile 20 suspension mg Example 2 3'-desamino-3'- (3-cyano-4-morpholinyl) doxorubicin 0.5 25 Sodium citrate Sodium carboxymethylcellulose fibrous 5.7 lulose (low viscosity grade) 2.0 Methyl para-hydroxybenzoate 1.5 30 Propyl p-hydroxybenzoate Water for injections up to 1 ml 0.2 Preparation B: Capsule mg 35 Example 2 10 Lactose 190 Preparation D ': Capsule preparation mg 40 Example 2 3'-desamino-3'- (3-cyano-4-morpholinyl) doxorubicin Lactose 1 199 45 The compounds of Example 2 and form- let the lake pass through a sieve, a poro-

mix well before filling into hard gelatine capsules so that each capsule contains 200 mg of mixed powder.

Claims (2)

A process for the preparation of compounds of the formula I wherein R is CO-CH ₃ or -CHOH-CH _{3 for} daunorubicin derivatives, or -CO-CH ₂ OH or -CHOH-CH ₂ OH for doxor bicin derivatives; characterized in that a compound of formula (III) wherein R is -CO-CH ₃ or -CO-CH ₂ OH- is reacted with 18 molar equivalents of 2,2'-oxoacetaldehyde (1) in a mixture of water and a polar organic solvent. Which is prepared in situ from 1,4-anhydroerythritol at room temperature, cleaved with sodium periodate in the presence of two molar equivalents of the alkali metal cyanoborohydride with respect to the starting material, and the corresponding basic morpholine derivatives of Cyano-morpholine analogs are obtained which are separated into basic and neutral fractions and the neutral fractions are chromatographed on a silica gel column eluting with dichloromethane: methanol (9: 1 by volume) to give the desired eluate in the first eluate. 3-cyano-morpholino - where R is -CO-2H ₃ or -CO-CH ₂ OH- and the following eluate is obtained, the corresponding 3-cyano-13-dihydro derivative of formula (I) - wherein R is -CHOH-CH ₃ or - CHOH-CH ₂ OH and the resulting product is purified if desired. 2. A process for preparing a pharmaceutical composition comprising mixing a compound of claim 1, wherein R is as defined in claim 1, with a pharmaceutically acceptable carrier or other excipient and converting it into a pharmaceutical composition.