Synthesis of phosphoramide mustard analogues of daunomycin and carminomycin.

New phosphoramide mustards (6-8) have been prepared from the antibiotics 2 and 3, and from 5. The mixture of cyclophosphamides could be separated by preparative layer and column chromatography. The assignments of configuration to the isomeric phosphoramidates was based on the magnetic anisotropy of the P = O bond. The synthesized compounds 6a,b-8a,b (mixture of isomers) were tested for inhibitory activity on the [3H]-thymidine incorporation into the DNA of tumor cells, using ovarian carcinoma cell line.

[Synthesis and mass-spectrometric analysis of N-cycloalkyl derivatives of carminomycin, daunorubicin and their analogs]. / Poluchenie i mass-spektrometricheskoe izuchenie N-tsikloalkil'nykh proizvodnykh karminomitsina, daunorubitsina i ikh analogov.

Condensation of carminomycin or daunorubicin with glutaric dialdehyde in the presence of NaBH3CN yielded 3'-deamino-3'-piperidinocarminomycin or 3'-deamino-3'-piperidinodaunorubicin and corresponding (13-R, S)-dihydroderivatives. To prepare similar derivatives of 14-hydroxycarminomycin or doxorubicin, 13-dimethylketals of 14-bromocarminomycin or 13-bromodaunorubicin were used in the reaction of reductive alkylation with glutaric or glycolic dialdehyde to give 3'-deamino-3'-piperidino- or 3'-deamino-3'-morpholino derivatives of 13-dimethylketals of 14-bromocarminomycin or daunorubicin, respectively. After deblocking and subsequent hydrolysis of these compounds 3'-deamino-3'-piperidino- and 3'-deamino-3'-morpholino derivatives of 13-hydroxycarminomycin or doxorubicin were prepared. Reduction of the antibiotic derivatives under mass spectrometry conditions was demonstrated.

[New derivatives of carminomycin and rubomycin]. / Novye proizvodnye karminomitsina i rubomitsina.

For preparing new semisynthetic analogs of anthracycline antibiotics, hydrolysis of 13-dimethylketals of 14-bromrubomycin and 14-brom-arminomycin in solution of diluted hydrochloric acid was studied. It was shown that such hydrolysis yielded 14-chlorrubomycin and 14-chlorcarminomycin. Conditions for separating the mixture of 14-chlor- and 14-bromrubomycins and the mixture of 14-chlor- and 14-bromcarminomycins by HPLC were developed. Interaction of 14-chlorine derivatives of rubomycin and carminomycin with potassium formate in the presence of the crown ether yielded 14-formyloxy derivatives of rubomycin and carminomycin. Interaction of rubomycin and carminomycin with formic acid in the presence of N-oxysuccinimide and dicyclohexylcarbodiimide resulted in formation of N-formyl derivatives of rubomycin and carminomycin.

[Synthesis and antitumor action of 14-N-substituted derivatives of rubomycin and karminomycin]. / Sintez i protivoopukholevoe deistvie nekotorykh 14-N-zameshchennykh proizvodnykh rubomitsina i karminomitsina.

The reaction of nucleophilic substitution of 14-bromine derivatives of carminomycin and rubomycin with respective nitrogen-containing heterocycles yielded six novel derivatives of carminomycin and rubomycin: 14-N-imidazolyl-carminomycin, 14-carminomycyl-N-pyridinium bromide, 14-carminomycyl-N-(3-aminocarbonyl)-pyridinium chloride, 14-rubomycyl-N-(3-amino-carbonyl)-pyridinium chloride, 14-N-succinimidocarminomycin and 14-N-succinimidorubomycin. In vitro and in vivo antitumor activity of the above derivatives and three other derivatives described earlier: 14-rubomycyl-N-pyridinium bromide, 14-N-imidazolylrubomycin and 14-N-phthalimidorubomycin was studied. It was shown that in vitro all the 9 semisynthetic derivatives had a lower (by 1.5-6 times) cytostatic action on murine lymphadenosis cells NK/LI as compared to the initial antibiotics. The in vivo experiments on mice revealed that by acute toxicity the rubomycin derivatives administered intravenously were close to rubomycin, whereas the toxicity of the analogous derivatives of carminomycin was 5-17 times lower. The in vivo experiments also showed that seven out of the nine 14-N-substituted derivatives of carminomycin and rubomycin were practically deprived of antitumor activity (strain LIO-1), while 14-carminomycyl-N-pyridinium bromide and 14-N-succinimidocarminomycin inhibited the tumor growth by 40-60 per cent.

The absolute structures of rubeomycins A and A1 (carminomycins II and III) and rubeomycins B and B1 (4-hydroxybaumycinols A1 and A2).

The absolute configurations of rubeomycins A and A1 (corresponding to carminomycins II and III) and rubeomycins B and B1 (corresponding to 4-hydroxybaumycinols A1 and A2), except at the C-1" position, were determined by comparison of the optical rotations and other spectral data of rubeomycin derivatives with those of daunomycin and L-(+)-lactic acid.

[Synthesis and antitumor properties of carminomycin 13-cyclohexylidenhydrazone]. / Sintez i protivoopukholevye svoistva 13-tsiklogeksilidengidrazona karminomitsina.

Carminomycin 13-cyclohexylidenhydrazone (CCH) was prepared by interaction of carminomycin 13-hydrazone with cyclohexane. The antiblastomic properties of CCH were studied on mice with transplantable tumors. The preparation was administered intravenously or orally. The studies showed a high antitumor activity of CCH. When CCH was administered intravenously to mice with lymphosarcoma LIO-1, the antitumor effect selectivity of it was practically equal to that of carminomycin. When used in doses equivalent by their toxicity to those of carminomycin, CCH had practically the same inhibitory effect on sarcoma 180 as carminomycin. When used orally in doses equivalent by their toxicity to those of carminomycin, CCH was more effective than carminomycin in treatment of mice with lymphosarcoma LIO-1, sarcoma 180 and lymphadenosis NK/Ly.

[Production of 14-substituted derivatives of carminomycin and rubomycin]. / Poluchenie 14-zameshchennykh proizvodnykh karminomitsina i rubomitsina.

14-Bromocarminomycin and 14-bromorubomycin were treated with alkali metal salts and nitrogen heterocycles to obtain 14-acetoxycarminomycin, 14-octamoylhydroxycarminomycin, 14-salicyloylhydroxycarminomycin, 14-salicyloylhydroxyrubomycin, 14-chinaldinoylhydroxyrubomycin and rubomycin 14-N-phthalimide, rubomycin 14-N-pyridinium bromide and 14-N-imidazolylrubomycin. It was shown that the reaction rate of the nucleophilic substitution in the acetone medium could be increased with the use of crown ethers of sodium iodide. Under such conditions 14-iodinecarminomycin and 14-iodinerubomycin, two intermediate products, partially reduced to the initial antibiotics, i.e. carminomycin and rubomycin. 14-Acetoxycarminomycin had the highest activity against Bac. mycoides, used as a test microbe. It amounted to 50 per cent, while the activity of the other derivatives did not exceed 25 per cent of the activity of the initial antibiotics.

[Synthesis and study of the antitumor properties of 13-hydrazone-substituted carminomycin]. / Sintez i izuchenie protivoopukholevykh svoistv zameshchennykh 13-gidrazonov karminomitsina.

13-Tret-butoxycarbonyl hydrazone (BOC hydrazone) of carminomycin was prepared by interaction of carminomycin with tret-butoxycarbonyl hydrazine. Interaction of carminomycin with N-amino-N'-methyl piperazine resulted in 13-(4-methyl piperazinyl) imine (MP imine) of carminomycin. BOC hydrazone and MP imine of carminomycin had a significant antiblastomic activity against lymphosarcoma LIO-1. Still, they had no advantages over carminomycin.

[Cytostatic action of semisynthetic derivatives of carminomycin]. / Izuchenie tsitostaticheskogo deistviia nekotorykh polusinteticheskikh proizvodnykh karminomitsina.

The cytostatic effect of some semi-synthetic derivatives of carminomycin on the tumor cells of mouse lymphadenosis (NK/Ly was studied in primary suspended cultures. A rapid method developed by the authors was used for determination of the drug cytostatic effect. The method is based on measuring the intensity of 3H-thimidine incorporation into the tumor cells. Simultaneously a method based on estimation of the increase of the total amount of the nucleic acids was used. No derivative superior to the initial drug by the level of the cytostatic effect on the tumor cells was found.

[Acute toxicity of the N-acyl derivatives of carminomycin and rubomycin]. / Izuchenie ostroi toksichnosti nekotorykh N-atsil'nykh proizvodnykh karminomitsina i rubomitsina.

Synthesis of 2 new N-acyl derivatives of carminomycin and rubomycin (N-L-leucylcarminomycin and N-sarcolysylrubomycin) is described. Acute toxicity of the new and 4 known N-acyl derivatives: N-acetylcarminomycin, N,L-alanylcarminomycin, N-D-phenylalanylcarminomycin and N-D-phenylalanylrubomycin was studied on albino mice. It was shown that the N-acyl derivatives of carminomycin and rubomycin had lower acute toxicity than the initial drugs. When added to blood serum in vitro N-D-phenylalanylcarminomycin and N-D-phenylalanylrubomycin induced precipitation. The carminomycin derivatives containing the residues of L-leucine and L-alanine were less toxic than the initial antibiotic, still they had a markedly pronounced retarded toxicity.

[Synthesis and properties of the N-ethyl derivatives of carminomycin and rubomycin]. / Sintez i svoistva N-étilproizvodnykh karminomitsina i rubomitsina.

N-Monoethyl derivatives of carminomycin, rubomycin, 13-dihydrocarminomycin and 13-dihydrorubomycin were synthesized by condensation of their amino groups with acetic aldehyde in the presence of sodium boron hydride. The respective N,N-diethyl derivatives of the antibiotics were formed as by-products of the reaction. New compounds such as N-ethylcarminomycin, N,N-diethylcarminomycin, N-ethyl-13-dihydrocarminomycin, N,N-diethyl-13-dihydrocarminomycin, N-ethylrubomycin and N-ethyl-13-dihydrorubomycin were synthesized. Antibacterial activity of N-ethyl- and N,N-diethyl derivatives of carminomycin and rubomycin determined with the use of Bac. mycoides as the test microbe was 40-50 per cent and that of N-ethyl- and N,N-diethyl-13-dihydro-derivatives was 15-30 per cent of the activity of the respective antibiotics, carminomycin and rubomycin.

[Synthesis, toxicity and antitumor action of carminomycin azine (carminazine)]. / Sintez, toksichnost' i protivoopukholevoe deistvie azina karminomitsina (karminazina).

Carminomycin azine designated as carminazine was prepared by condensation of carminomycin with hydrazine hydrate. It was shown in the experiments on mice that the toxicity of carminazine was 2 and 7 times lower than that of carminomycin on its intravenous and oral administration respectively. The effect of both drugs on hemopoiesis of the mice was similar. As regards the selectivity of the antitumor effect on lymphosarcoma, strain L10-1, carminazine was inferior to carminomycin.

Synthesis of 2'-deoxy-L-fucopyranosylcarminomycinone and -epsilon-pyrromycinone as well as 2'-deoxy-D-erythro-pentopyranosyldaunomycinone, -carminomycinone, and -epsilon-pyrromycinone.

Treatment of di-O-acetyl-2-deoxy-L-fucopyranosyl bromide with carminomycinone and epsilon-pyrromycinone in the presence of mercuric bromide and mercuric cyanide afforded 3',4'-diO-acetyl-2'-deoxy-L-fucopyranosylcarminomycinone and -epsilon-pyrromycinone. Similarly, when di-O-acetyl-2-deoxy-D-erythrho-pentopyranosyl chloride was treated with daunomycinone, carminomycinone and epsilon-pyrromycinone, the di-O-acetyl derivatives of the anthracyclinone glycosides were obtained. Deacetylation of the previous acetates with sodium methoxide afforded 2'-deoxy-L-fucopyranosylcarminomycinone and -epsilon-pyrromycinone, as well as 2'-deoxy-D-erythro-pentopyranosyldaunomycinone, -carminomycinone, and -epsilon-pyrromycinone. 2'-Deoxy-L-fucopyranosylcarminomycinone was found to be more active than carminomycin at higher dosages on L1210.

[Synthesis and properties of the N-acyl derivatives of carminomycin and rubomycin]. / Sintez i svoistva nekotorykh N-atsil'nykh proizvodnykh karminomitsina i rubomitsina.

Synthesis of N-acetylcarminomycin (III) is performed for the first time and the method of selective aminoacylation of 3'-NH2 group of carminomycin (I) and rubomycin (II) is elaborated. The method is based on interaction of N alpha-protected amino acid activated with respect to the carboxyl group with one of the antibiotics followed by removal of the protective group under mild conditions. O-Nitrophenylsulphenyl group (NPS) is used as the protective agent. In case of rubomycin (II), N alpha-NPS-amino acid is attached with the carbodiimide method. In case of carminomycin (I) the similar reaction proceeds inconsistently and carminomycin is acylated with the method of activated ethers and the use of N-hydroxysuccinimide ether of N alpha-NPS-protected amino acid. The following substances were prepared: N-(N alpha-NPS-D-phenylalanyl)-carminomycin (IVa), n-(n alpha-NPIa) and N-(N alpha-NPS-L-alanyl)-rubomycin (VIIa). Removal of NPS-group is performed under mild conditions (2 equiv. HCl in acetone, 20 degrees C, 2 min) without significant destruction of the antibiotic glucoside bond and results in formation of respective water-soluble hydrochlorides of N-aminoacyl derivatives of carminomycin and rubomycin (IV-VII). Teh structures of the new compounds are confirmed by the analytical and spectral data (Rf, [alpha] D, IR-, UV- and VO-, PMR-spectra, elementary analysis) and their chemical transformations. Antimicrobial activity of N-acetylcarminomycin (III) and water-soluble hydrochlorides of carminomycin and rubomycin derivatives (IV-VII) against Bas. mycoides is 5-10% of that of the respective initial antibiotics (I) and (II).

Synthesis and antitumor properties of 7-deoxy-7-[(cis- and trans-3-aminocyclohexane)thio]carminomycinone.

The synthesis of analogues of carminomycin in which the daunosamine group has been replaced by (cis- and trans-3-aminocyclohexane)thio moieties is described. The new compounds were found to exhibit none of the antitumor or antibiotic activity associated with carminomycin.

The synthesis of epsilon-rhodomycinone- and carminomycin-11-methyl ethers.

The conversion of epsilon-rhodomycinone to its 11-methyl ether via selective hydrolysis of the 4,6,7,11-tetraacetate is described. This series of reactions was used as a model for the conversion of carminomycin to its 11-methyl ether. The anti-tumor activity of the latter compound was less than that of both carminomycin and its 4-methyl ether (daunomycin).

[Synthesis and properties of the 14-amino derivatives of daunorubicin and carminomycin]. / Sintez i svoistva 14-aminoproizvodnykh daunorubitsina i karminomitsina.

Interaction of 14-bromine derivatives of daunorubicin and carminomycin, as well as their aglycones with secondary amines, such as piperidine, N-methylpiperazine and morpholine was studied with a purpose of preparing new potentially antitumor antibiotics. It was found that reaction in acetone or dioxan at a temperature of 50--60 degrees C resulted in formation of 14-amino derivatives of the respective bromides. 14-Piperidinyl- and 14-(N-methylpiperazinyl)-daunorubicin, 14-piperidinyl-daunorubicinon and 14-piperidinyl-carminomycinon were prepared. The structure of the new substances was confirmed by the IR, UV and NMR spectra. The antimicrobial activity of 14-amino derivatives amounted to 10--35 per cent of the activity of the initial antibiotics, i.e. daunorubicin and carminomycin.

[Production and antitumor properties of carminazone]. / Poluchenie i protivoopukholevye svoistva karminazona.

Karminazon (13-benzoylhydrazon) was prepared by condensation of karminomycin with benzoylhydrazine. In its intravenous use in the treatment of mice with lymphosarcoma L10-1 karminazon was less toxic and had lower antitumor activity than karminomycin. Karminazon had a lower selective antitumor activity with respect to lymphosarcoma than karminomycin.

Preparation and biological evaluation of 4-O-demethyldaunorubicin (carminomycin I) and of its 13-dihydro derivative.

A mutant strain of Streptomyces peucetius produced an anthracycline antibiotic whose structure has been established to be 4-O-demethyl-13-dihydrodaunorubicin (4), by application of spectroscopic methods and chemical degradation. A new synthesis of 4-O-demethyl-daunorubicin (carminomycin I, 2) starting from daunomycinone, together with the comparison of the antitumor activity of the anthracycline glycosides 2 and 4 are also reported.