CS268514B2 - Method of daunorubicine's and doxorubicine's nes 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

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

Doxorubicin (adriamycin) described in U.S. Patent No. 3,590,028 (F. Arcamone et al.) Is probably the most potent anticancer agent currently in use. Along with daunorubicin, it is used to treat a variety of tumors as well as leukemia. However, some patients with these tumors, especially some severe cancers such as colon and melanoma, do not respond favorably to this treatment. In addition, some patients undergo untreatable cardiac damage with prolonged treatment, which can itself lead to death. Therefore, it would be desirable to find analogous substances with a broader effect and 8 lower cardiac muscle toxicity. Up to now, some active novel analogs, such as compounds active against murine P388, are two lipophilic derivatives of AD32 and Κ, d-dibenzyldaunorubicin, but high doses are required and do not affect 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 disclosed in Cancer Chemotherapy, Doxorubicin (Federico Arcav US David W. Henry Adriamycin®, ACS Symposium Series, American Chemical Society, 1976, pp. 15-57) and in mone, Academic Prese, 1981). AD32 was disclosed in U.S. Patent No. 4,035,566, issued Jul. 12, 1977

5-Iminodaunorubicin has been described in U.S. Pat

1978 (David V. Henry and George L. Tong) lower side effects, 5-ininodoxorubicin has good efficacy, but higher doses are needed.

A group of new daunorubicin and doxorubicin derivatives has now been found. These compounds can be expressed

δ. 4

5-Iminodaunorubicin has

No. 109,076 of August 22, the same efficacy in Formula I

R

OH

(AND)

CS 268 514 B2 where

H is CO-CH3 or CHOH-CH3 for daunorubicin derivatives a

CO-CH ₂ OH or CHOH-CH ₂ OH in the case of doxorubicin derivatives,

A is cyano or hydrogen.

When A represents a hydrogen atom, the compounds may also exist in the form of addition salts and acids. These eoli also belong to the field of the invention.

The compounds of the formula I can be obtained by reacting a daunorubin derivative of the formula

where

R is -CO-CH 2, -CO-CH 2 -OH, -CHOH-CH 2 or -CHOH-CH ₂ OB, 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 obtained compounds are then treated with an excess of alcoholic ammonia solution to provide compounds of formula I which are effective against tumors. These substances seem to combine both advantages of high anti-inflammatory activity and the possibility of using low doses. For this reason, there are also lower side effects, for example cardiotoxicity compared to known compounds.

The compounds produced by the process of the present invention can be formulated as pharmaceuticals

CS 268 513 B2

means for the treatment of tumors in the usual way:

Morpholinyl derivatives of iminodaunorubicin and iminodoxorubicin and cyanmorpholinyl derivatives, iminodaunorubicin and iminodoxorubicin These are shown in Table I.

Table I

AND fi the name of the compound H CO-CH 3'-Deamino-3 '- (N-morpholinyl) -5-iminodaunorubicin and its pharmaceutically acceptable salts H саон-сн ₃ 3'-Deamino-3'-morpholinyl) -13-dihydro-5-iminodaunor- bicin and its pharmaceutically acceptable salts H co-ch ₂ oh 3'-Deamino-3,4'-morpholinyl) -5-iminodoxorubicin and its pharmaceutically acceptable salts H CHOH-CH ₂ OH 3-Deamino-3- (4 'morpholinyl) -13-dihydro-5-iminodoxorubicin and its pharmaceutically acceptable salts CN CO-CH ₃ 3-Deamino-3 * - (3'-cyan-4 * -morpholinyl) -5-iminodaunorubicin CN -CHOH-CH ₃ 3'-Deamino-3'- (3-cyano-4-morpholinyl) -13-dihydro-5-iminodaunorubicin CN CO-CH ₂ OH 3 deamino-3 '3-cyano-4 ^{M-morpholinyl)} -5-iminodoxorubicin CN CHOH-CH ₂ OH 3'-Deamino-3 ^z - (3'-cyan-4-morpholinyl) -13-dihydro-5-imi

nodoxorubicin

One of these compounds is most preferred for its very good activity against tumors. The substance is:

* -deamino-3 * - ( ^4M- morpholinyl) -5-iminodoxorubicin

The first four compounds listed in Table I may be used in the form of free compounds or in the form of salts with pharmaceutically acceptable bases. The addition salts have the advantage of being soluble in water and water-containing solvents, for example in a mixture of water and an alkanol, or in a mixture of water and an alkanediol. Examples of such mixed solvents are a mixture of water and propylene glycol, water and ethanol, water and ethylene glycol, aqueous sodium chloride solution and various injectable media containing water and the like. Free Substances They are soluble in less polar organic solvents such as chloroform, methylene chloride, chloroform-methanol mixtures and the like. They can also be used in the form of a suspension.

The salts can also be free acid addition products with a pharmaceutically acceptable acid. As used herein, the term is a non-toxic acid and is commonly used in pharmaceutical formulations. Examples of such acids are inorganic acids such as hydrochloric, hydrobromic, sulfuric and phosphoric acids and organic acids such as carboxylic acids such as acetic, glycolic, maleic, malic, hydroxynaleic, tartaric, citric and salicylic acids as well as organosulfonic acids such as methanesulfonic acid; p-toluenesulfonic acid. Mixtures of two or more acids and mixtures of one or more free substances or addition salts may also be used. Hydrochlorides and hydrobromides are most preferred.

CS 268 513 B2

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 3 is reacted under conditions of reductive alkylation with 2,2'-oxodiacetaldehyde of the formula = CH CH = O

CH ₂ - O - CH ₂

This alkylation yields a mixed product containing four major components. In the case of the use of daunorubicin, these components are the following four compounds:

'-deamino-3 * - ( ^4H- morpholinyl) daunorubicin, ^from -deamino-3 * - (4'-morpholinyl)' 13-dihydrodaunorubicin, 3'-deamino-3 * - (3-cyano-4-morpholinyl) daunorubicin and ^z- deamino-3 * - (3-cyano-4 ** -morpholinyl) -13-dihydrodaunorubicin.

In the case of doxorubicin, these components are the following four compounds:

'deamino-3- ⁽⁴ watts morpholinyl) doxorubicin, -.

* -deamino-3 '- ( ^4M- morpholinyl) -13-dihydrodoxorubicin, * -deamino-3- (3-cyano-4'-morpholinyl) doxorubicin and ^z -deamino-3 * - (3-cyano-4- morpholinyl) -13-dihydrodoxorubicin.

2,2'-Oxydiacetaldehyde can be obtained by hydrolyzing 2,2'-oxydiacetaldehyde bis (diethyl acetal) in an acidic medium. The starting material can be represented by the formula (2t-O) ₂ -CH CH- (O-Et) ₂ *!

The CH ₂ · -O-CH ₂ process is carried out according to the method of Belgian patent specification 655 436 (Pield et al.), The same substance can also be obtained by cleavage of 1,4-anhydroerythritol of the formula

HO - CH ------ - CH - OH

CH ₂ - 0 - ₂ according to Wagg et al., Carbohydrate Researoh, 2, 299 (1968) and Greenberg et al. Carbohydrate Research, 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 reaction is very easy to carry out and is usually completed within 1 hour or even shorter at room temperature. Reductive alkylation is explained in more detail in the following examples and is also used in U.S. Patent No. 4,301,277 and described in J. Medicinal Chem., Pp. 18-24 (19-2).

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 cyanoacin free materials from acid-insoluble and cyano-containing materials. The resulting material pairs are possible

CS 268 514 B2 by liquid chromatography.

and directly from the isolated 5-oxo compounds

Chem., 24, 669 (1981). When chromatographed on individual substances, for example by column chromatography, thin layer chromatography or high pressure

The 5-imino compounds can be readily prepared using the method of J. Medicinal By contacting this process, 5-oxo materials are contacted with an excess of alcoholic ammonia solution at a low to moderate elevated temperature, for example at -25 to + 25 ° C at 0 ° C. , 5 to 100 hours. In the case of 3'-deamino-3 * - (4-norfolinyl) -doxorubicin and 3'-deamino-3 * '(3'-cyano-4-morpholinyl) -doxorubicin, it is necessary to protect the hydroxyl group on the carbon atom at position 14 from the action ammonia. Any protecting group that is labile in a mild acidic environment can also be used for this purpose. Because of its wide range of uses in pharmaceutical chemistry, the preferred protecting group is the methoxytrityl group. The trityl function can be introduced by treating 3 * -deamino-3 * - ( ^4H- aorpholinyl) -doxorubicin or 3 * -deamino-3 * - (3-cyano-4-morpholinyl) -doxorubicin with an excess of p -anisylchlorodiphenylmethane at room temperature. After the reaction with ammonia is complete, the hydroxyl group at the 14-position can be regenerated by treatment with an acid such as acetic acid or a cold aqueous solution of trifluoroacetic acid. · '

The invention will be illustrated by the following examples.

Example 1

Preparation of 3'-deamino-3 '- (4'-morpholinyl) -5-amino-oxorubicin

А. To a solution of 0.396 g of 3 * -deamino-3 * - (4-inorpholinyl) doxorubicin, in 5 ml of anhydrous pyridine, 0.990 g of p-anisylchlorodiphenylmethane is added. The mixture is allowed to react at room temperature in the dark for about 2 days. The solution is then cooled with a mixture of water and ice and 0.5 ml of methanol are added. The mixture is stirred for a further 2 hours, then 50 ml of dilute sodium bicarbonate are added and the mixture is extracted with methylene chloride. The extracts are concentrated to a gummy residue which is dissolved in toluene, evaporated, dissolved in methylene chloride and precipitated by the slow addition of petroleum ether, b.p. 35-60 ° C. The precipitate was collected, redissolved in methylene chloride and precipitated with a mixture of petroleum ether and diethyl ether to give 14-O-p-anisyldiphenylmethyl-3 * -deamino-3 * - (4-norfolinyl) doxorubicin (III) as an amorphous solid in 94% yield.

CS 268 514 32 ·

This material was identified by NMR analysis at 90 MHz in CHCl 3

B. A solution of 0.532 g of 14-O-p-anisyldiphenylmethyl-3 ' -deamino-3 '-( 4-morpholinyl) doxorubinating in 10 ml of methylene chloride was added to 30 ml of methanol saturated with ammonia at 0 [deg.] C. The mixture was stirred at 0 ° C for 1 hour and then allowed to stand at 3 ° C for 27 hours. The solvent was evaporated and the residue was dissolved in methylene chloride / methanol (4: 1) and evaporated again. The procedure is repeated twice, then the solid obtained is dissolved in methylene chloride, filtered through a Cell tu layer, evaporated, the residue is dissolved in a 1: 2 mixture of methylene chloride and methanol, and the solution is again evaporated to dryness and dried, 0.52 g of a violet residue is obtained in 97% yield.

C. Dissolve the residue from Step 3 in 2 ml of tethylene chloride and apply the solution to a 1.5 x 40 cm silica gel column, eluting the column with 50 ml of methylene chloride and then with a mixture of methylene chloride and methanol, first 150 ml of a 98: 2 mixture. , 500 ml of 87: 3, 100 ml of 95: 5, 100 ml of 7 and 200 ml of 90: Ю. First, 565 ml of eluate is collected, then a 335 ml fraction is collected, filtered and evaporated to contain 59.9% of the loaded sample as a uniform material. This material was identified as 14-O-p-anisyldiphenylmethyl-3'-deamino-3 '- (4-morpholinyl) -5-iminodoxorubicin by NMR analysis at 90 MHz. ’

D. 0.341 E of the product from Step C was dissolved in 20 cl of 80% acetic acid and stirred in the dark for 7 hours. The solution was diluted with 50 mL of water and extracted three times with chloroform. The aqueous phase containing the desired product is lyophilized in the dark to give 0.294 b of a solid. This solid ae was dissolved in 0.1 N acetic acid. The solution was washed with chloroform, basified with sodium bicarbonate and extracted with chloroform. The desired product is taken up in an organic phase which is washed, dried, filtered and evaporated to a residue. This residue is dissolved in a 1: 10 mixture of chloroform and methanol, the solution is evaporated and the residue is dried, yielding 0.228 g of 3'-deamino- 3 * - (4-morpholinyl) -5-iminodoxorubicin. The identity of this material was verified by NMR analysis at 300 MHz and elemental analysis.

Example 2

Process for the production of acid addition salts

The free base of the product of Example 1 is suspended in 20 ml of water. The mixture was stirred while 3.2 ml of 0.1 N hydrochloric acid was slowly added to pH 4.5. The suspended solid dissolves gradually. The solution is lyophilized in the dark to give 3 deamino-3 '- ⁽⁴ watts morpholinyl) -5-iminodoxorubicinhydrochloridu a purity of 97% with HPLC analysis.

Analysis for .HCl.2 h ₂ o calculated: C: 54.35, H: 6.03, Cl ': 5.17, N: 4.09% found: C: 54.20, H: 5.96, Cl: 4,33, N: 4.03% At clade 3

Preparation and isolation of 3 * -deamino-3 * - (4'-morpholinyl) -5-imino-13-dihydrodoxorubicin.

: .1

CS 268 514 В2

A solution of 0.186 g 3 deamino-3 '- ⁽⁴ watts morpholinyl) -13-dihydrodoxorib Cause, in 6 ml of methylene chloride and methanol in a ratio of 1: 1 was added к 20 ml of methanol saturated with ammonia at 0 ° C. The mixture was stirred for 1 hour and then allowed to stand at 3 ° C for 27 hours. The mixture was evaporated and the residue was dissolved in methylene chloride / methanol (4: 1) and the procedure repeated three times until complete removal of ammonia. The resulting residue was purified by silica gel thin layer chromatography (mm.times.20.times.20 cm) using a 9: 1 mixture of chloroform and methanol as solvent. Bands containing substantially pure 3 * -deamino-3 * - (4 ^M) (morpholinyl) -5-imino-13-dihydrodoxorubicin substances as possible

B. The free material was collected, washed and the eluate was evaporated, yielding 0.139 g free of K 2 M analysis at 300 MHz. .

from Step A was converted to the hydrochloride by the method of Example 2. The purity of the hydrochloride was shown to be 97-98% by HPLC. *

Analysis for θ3ΐ ^ 38 ^ 2θ11 ^ θ ^ * 2H ₂ O

Calculated: C: 54.19, H: 6.31, Cl: found: C: 54.17, H: 5.95, Cl:

5.16,

4,88,

N: 4.08%

N: 3.87%

Example 4

Preparation of 3'-deamino-3 '- ⁽³ watts cyano-4''morpholinyl) -5-iminodaunorubicin

A solution of 0.031 g ^of 3 deamino-3 '- (3-cyano-4 * ^{H-morpholinyl)} daunorubicin in 1.0 mL of methylene chloride was added 5 ml к methanol saturated with ammonia at 0 ° C. The mixture was stirred for 30 minutes and then stored at 3 ° C for 45 hours. The reaction product is evaporated to dryness and the residue is dissolved in 5 ml of a 19: 1 mixture of chloroform and methanol, then the solution is evaporated. This step is repeated once more. Dissolve the residue in a methanol / chloroform mixture and apply to a thin layer of silica gel, the plate having a size of 2 mm x 20 x 20 cm, using a 9: 1 mixture of chloroform as the solvent. Mass spectroscopy can be verified that the resulting substance is a 3 ' -deamino-3 ^of - (3-cyano-4 * ^{H-morpholinyl)} -5-iminodaunorubicin.

Mass spectrum in the form of trimethylsilyl derivative (TMS): m / e 882 / m (TM6) .delta., 810 (M) (IMSI4 -HCN). The mass spectrum had a baseline (HCM) at m / e 27 · Chromatography on thin layer of silica gel using a 19: 1 mixture of chloroform and ethanol 0.32.

Example 5

The procedure of Example 1 was again repeated using deamino-3 ^{of from} 3 - (3-cyano-4 ^M ** - morpholinyl) doxorubicin, instead of 3 ' -deamino-3 '- (4-morpholinyl) doxorubicin. The sequence of deprotection, amination and deprotection of Example 1 results in 3'-deamino-3 ^from - (3'-cyano- ^4H- morpholinyl) -5-iminodoxorubicin as the final product.

The reaction can be described in more detail as follows:

А. К solution of 0.241 g ^of 3 deamino-3 '- (3-cyano-4-morpholinyl) doxorubicin in 4 ml of anhydrous pyridine was added 0.587 g of p-anisylchlordifenylmethanu. The solution is stirred at room temperature in the dark for 44 hours, then the reaction mixture is cooled, diluted with 0.5 ml of methanol, stirred at room temperature for 3 hours, then added to 50 ml of dilute solution.

The mixture was extracted with methylene chloride. The extracts were evaporated and the residue dissolved bs vs 3 ml of methylene chloride and precipitated by slowly adding 40 ml disthylesteru give in 97% yield 0.333 g p-14-0 anisyldifenylmethyl-3'-deamino-3 '- ^(3H - cyan-4 ⁽ morpholinyl) doxorubicin.

./ =

NMB 3 pektrum at 9 < 3 MHz in CDCl3 13, 84 (with, 6 — OH), 12.99 (with, 11-OH), 7.82 (and, 1-H), 6.70 to 7.75 (m, 2-H, 3-H 5.42 (bs , i'-h) 5.08 (bs , 7-H), 4.45 (bs 2, 14-H ₂ ) 4.19 (with, 9-OH), 4.00 (з> осн ₃ ), 3.79 (with, OCH ^), 3.30 to 4.15 (m, 4'-H, 5'-H 1.60 to 3.10 (m, 2 '-H _2, 8-H ₂ 1.13 (and, 6'-h ₃ ).

2-H ₂ , 3-H, 6-H ₂ ), 5 ^w -H ₂ , 10-H ₂ , Z ₁ -N),

B. Kiss 0.369 g of 14-O-p-anisyldiphenylmethyl-3 ' -deamino-3 &apos;-( 3 &apos; -cyano-4-morpholinyl) doxorubicin in 8 ml of methylene chloride was added to 25 ml of methanol saturated with ammonia at 0 ° C. The mixture was stirred at the same temperature for 1 hour and then allowed to stand at 3 ° C for 26 hours. The mixture was then evaporated to remove ammonia to give 0.376 g of a yellow residue. ·

C. The residue from step V in 1.5 ml of methylene chloride is applied to the top of a 1.5 x 20 cm silica gel column with a particle diameter of 200 to 400 mesh and the column is washed with 50 ml of methylene chloride and then with a mixture of methylene chloride and methanol, first 200 ml. 99: 1 mixtures, 300 ml 98: 2 mixtures, 100 ml 97: 3 mixtures, 100 ml 95: 5 mixtures and 200 ml 90: 10 mixtures. Take 360 ml of the starting eluate, then the 125 ml fractions were evaporated to give 0.203 g of 14-O-p-anisyldiphenylmethyl-3'-deamino-3 '- (3'-cyano-4-morpholinyl) -5-iminodoxorubicin.

D. The 0.15 g of the residue from Step C was cooled to 0 ° C and dissolved in 8 ml of ice-cooled 50% trifluoroacetic acid. The solution was stirred at 0 ° C for two minutes and then poured into 100 ml of a mixture of water and ice. The aqueous mixture was extracted with chloroform (4 x 10 mL), the extracts were combined, washed with dilute sodium bicarbonate and water, dried over sodium sulfate, filtered through a pad of Celite and evaporated. The residue is dissolved in 3 ml of a 4: 1 mixture of chloroform and methanol. While stirring, 25 ml of ether are added to the solution and the resulting precipitate is collected to give 0.093 g of 3'-amino-3- (3-cyano-). 4-morpholinyl) -5-iminodoxorubicin.

HPLC analysis at 300 MHz indicated that the material was a mixture of diastereoisomers. High pressure liquid chromatography on a Waters Badial-Pak C-18 column in 0.05 M citrate buffer pH 4 / methanol 40:60 showed peaks at 18.4 minutes and 25.0 minutes at 69:31 The NMB spectrum of this product at 300 MHz has two resonances for the protons 1-H, 2-H, 3-H, fH, 7-H, 14-H ₂ , 9-OH, OCH 2, 10A-H and '- Hj.

NMB spectrum at 300 MHz in CDCl3:

15.61

13.74

9.27 (s, 11-OH), (d, 6-OH), (d, NH),

CS 268 513 B2

8,21, 8.19 (2d, 1H) 7.73, 7.72 (3t, 2-H), 7.33, 7.32 (2d, 3-H), 5.77, 5.72 (2d, 1'-H), 5,41, 5.38 (2a, 7-H), 4,79, 4.77 (2з, 14-H) 4,72, 4.66 (2b, 9-OH), 4,15, 4.14 (2s, OCH 2), 4.04 (in, 5'-H), 3.97 (in, 3-Н, 2 3.75 (in, 6-Н ₂ , 3.59 (а, 2А-Н), 3.23 (d, 10В-Н), 3.03 (а, 10А-Н, 2.72 (in, 5-Н ₂ ) » 2.33 (а, 8В-Н), 2.14 (в, 8А-Е), 1.85 (in, 2'-Н ₂ ), 1,38, 1.37 (2d, 6'-N ₃ )

The ultraviolet (Vis) spectrum in methanol has maxima at 221 mm (Λ 31 000), 252 (32 900), 307 (7 110),

520 hump (9,110), 551 (17,400), 592 (20,700).

DCI-MS m / e 638 (M + H) 611 (M + H-NCH).

Calculate: C: 53.62, H: 5.69, N: 6.41%, found: C: 58.79, E: 5.47, N: 6.30%,

Example 6

The procedure of Example 3 was repeated a total of four times, each time using an equivalent molar amount of the other starting material instead of 3'-deamino-3,4-morpholinyl) 13-dihydrodoxorubicin. In the first case, 3'-deamino-3 '- (3'-cyano-4-morpholinyl) -13-dihydrodoxorubicin is used and 3-deamino-3- ( ^3H- cyano-4-morpholinyl) is obtained as the product. ) -5-Imino-13-dihydrodoxorubicin.

Mass spectrum (in the form of the trimethylsilyl derivative (DIS): m / e 900 (M (TMS) 2 - HCN). On silica gel chromatography using a 9: 1 mixture of chloroform and methanol, R fp 0.24.

The compounds of the present invention can be used in mammals against cancer. This activity has been verified by invivo and in vitro experiments. In one in vivo experiment conducted according to Cancer Chemotherapy jReports, Kational Cancer Institute, 3, δ. 2, Part 3, September 1972, a healthy amount of P-388 lymphocytic leukemia was intraperitoneally inoculated into healthy mice. After inoculation, mice were treated 5, 9, and 13 days with varying amounts of compounds produced by the method of the invention. Controls were untreated mice, other mice were treated for comparison of daunorubicin or doxorubicin 3 deamino-3 - (4 '-morpholinyl) daunorubicin or 3 deamino-3 - ⁽⁴ watts morpholinyl) -13-dihydrodaunorubicin according to U.S. Pat. 4,301,277 or 3 deamino-3 - ⁽⁴ watts methoxy-l-piperidinyl) daunorubicin or the 13-dihydroekvivalent according to U.S. Pat. No. 4,314,054.

CS 268 513 B2

The mean survival time in the various treated mice was determined and compared to 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. improvement.

Table A

Activity against Leukemis P-388 leukemia in mouse L-1210 cells.

Compound · NSC No. ^x

-deamino-3- (4-morpholinyl) -13-dihydro-

-5-imidodoxorubicin 355. »465

For comparison:

daunorubicin 82, 151%

doxorubicin 123, 127

Survival time Ji T / C Optimal dose (q4d 5, 9, 13) mg / kg inhibition of ^ED 50 > DNA synthesis / UM RNA 161 50 > 100 24 130 8 0.66 0.33 130 8 1.5 0.58

^x NSC # = National Sorvice Center of the US National Cancer Institute.

Table A - continued

compound NSC Ž. ^X Survival time £ T / C Optimal dose (q4d 5, 9, 13) mg / kg inhibition of ^ED 50 '/ ^wM DNA RNA synthesis 3 -deanino-3 - (4-morpholinyl) daunorubicin 327, 451 166 0.2 0.76 0.10 3-deamino-3 - (4-morpholinyl) - -13-dihydrodaunorubicin 327, 450 132 0.2 2,2 0,67

CS 268 513 B2

Additional in vivo tests in mice against P-388 and L-121O leukemia and B-16 melanoma were performed according to the above-mentioned Cancer Chemotherapy Reporte 1972. Various routes of administration were followed, for example intraporitoneally, intravenously and orally, the results are shown in the following B.

Table В

Antitumor efficacy in mice at optimal dose% T / C (mg / kg) doxorubicin 355277 (1)

NSC δ. 123127 ^{* х} ip 388

d, 1, ip 252 (7.5)> 193 (2) d, 1, iv 185 (10) 166 (2.5)

Explanatory notes to Table B:

^x known substances for comparison (1) 3 * -deamino-3 * - (4-morpholinyl) -5-iminodoxorubicin

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. Particularly preferred is parenteral administration, especially intravenous administration. The doses and amounts as described above can be used to improve leukemia or any other type of cancer against which they are effective. For example, in the treatment of small test animals, doses of 0.0010 to 25 mg / kg per day may be used to improve leukemia. Typically, the dose of the compounds produced by the method of the invention is about 1/20 to 1/200 of the parent compounds. One dose may be administered every 2 to 7 days, but 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 which resolves the compound and facilitates administration. It is known that this carrier, like the resulting formulation, must be sterile and not toxic.

For oral administration, the carrier used as a diluent may be 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 known manner.

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 aqueous suspension for injection may be obtained from the following mixture:

ingredient · · mg

CS 268 514 B2 the compound of Example 1, 4, 5 in the form of a suspendable powder 3 sodium citrate 5.7 sodium carboxymethylcellulose (low viscosity) 2.0 methyl parahydroxybenzoate. .1,5 propyl parahydroxybenzoate G,

The above mixture is made up to 1.0 ml with water suitable for injection.

Means В. A sterile solution in an aqueous carrier for injection may be obtained from the following mixture:

Component of Example 24 sodium citrate * 5.7 carboxymethylcellulose sodium (low viscosity) 2.0 methyl parahydrobenzoate1.5 propylparahydroxybenzoate 0.2

The resulting mixture was made up to 1.0 ml with water for injection.

The compounds of the present invention may be administered, for example, in the form of tablets or capsules.

Composition C

Tablets can be made from the following mixture:

mg

5.0

51.5

2.5

1.0 compound of Example 2 lactose dried corn starch gelatin magnesium stearate

The compound of Example 2 is comminuted into a powder which is passed through a sieve and mixed with lactose and 30 mg of corn starch, which are also passed through a sieve.

The resulting powder is mixed in a warm gelatin solution, which is prepared by mixing gelatin with water and heating to form a solution having a concentration of 10% by weight. The resulting mass is passed through a sieve and the wet granule is dried at 40 ° C.

The dried granules are passed through a sieve, the rest of the starch and the magnesium stearate are added, and the mixture is thoroughly mixed.

Then the resulting mixture is compressed into 150 mg tablets.

CS 268 513 B2

Composition D

The capsules can be made from the following mixture:

Compound of Example 310 lactose190

The compound of Example 3 and the lactose are passed through a sieve and the powder mixture is mixed well, then filled into hard gelatin capsules of suitable size by inserting 200 g of the obtained powder material into each capsule.

Claims (1)

A process for the preparation of the novel moramoline derivatives of daunorubicin and doxorubicin of the formula I wherein the group is CO-CH 2, CHOH-CH 2, CO-CH 2 OH and CHOH-CH 2 OH and A is CN or hydrogen, characterized in that it is present in the compound of formula (i) i í and CS 268 514 B2 where R and A are as defined above, first when the compound contains a hydroxyl group at the 14-position, it provides a protecting group removable in a slightly acidic environment such as a p-methoxytrityl group, and then reacted with an excess of alcoholic ammonia solution at -25 to + 25 ° C, then optionally removing said protecting group with acetic acid or trifluoroacetic acid at room temperature and isolating and purifying the resulting material.