Synthesis and cytostatic properties of daunorubicin derivatives, containing N-phenylthiourea or N-ethylthiourea moieties in the 3'-position.

A series of phenylthiourea and ethylthiourea derivatives of daunorubicin and its congeners was prepared by reaction of the 3'-amino group of the antibiotic with phenylisothiocyanate or ethylisothiocyanate. S-Methylation yielded S-methylisothiouromium salts which when reacted with amines resulted in an intramolecular cyclization with the participation of the neighboring 4'-OH group. The structures and predominant conformations of the thiourea derivatives and daunorubicino(3'-N,4'-O-d)oxazolines were determined by 1H and 13C NMR. Cytostatic activities of the thiourea and oxazoline derivatives were compared with the cytostatic activities of N-methylurea and N-methyl-N-nitrosourea containing daunorubicin and its congeners. Carminomycin derivatives were endowed with the highest cytostatic activity.

[Establishing the structure of anthracyclinones produced by Streptomyces griseoruber]. / Ustanovlenie stroeniia antratsiklinonov, produtsiruemykh Streptomyces griseoruber.

Streptomyces grisoruber strain 1618-306 produces three types of anthracycline antibiotics, derivatives of epsilon-pyrromycinone (methyl (7S, 9R, 10R)-9-ethyl-5,7,8,9,10,12-hexahydro-1,4,6,7,9-pentahydroxy-5,12-di oxo-10- naphthacenecarboxylate), epsilon-1-hydroxyauramycinone and epsilon-1-hydroxysulfurmycinone, differing in C-9 substituent in D ring of anthracyclines (Et, Met or CH2COCH3, respectively). Besides 7-O-glycosides of these aglycones, complex of antibiotics contains corresponding 7-deoxy- and 7,8,9,10-bisanhydroanthracyclinones.

[14-O-hemiesters and 13-hydrazones of anthracyline antibiotics of the daunorubicin series. Synthesis and cytostatic activity with respect to tumor cells sensitive or resistant to doxorubicin]. / 14-O-gemiéfiry i 13-gidrazony antratsiklinovykh antibiotikov riada daunorubitsina. Sintez i tsitostaticheskaia aktivnost' v otnoshenii opukholevykh kletok, chuvstvitel'nykh ili ustoichivykh k doksorubitsinu.

Doxorubicin and 14-hydroxycarminomycin 14-O-hemiadipates and 14-O-hemipimelates, synthesized from 14-bromo derivatives of daunorubicin and carminomycin and monosodium adipate and pimelate, were converted to the corresponding N-trifluoroacetylated compounds. 13-(4-Methylpiperazine-1-yl)imino derivatives of the anthracycline antibiotics were also obtained. The cytostatic activity of the compounds synthesized was studied using a panel of human and animal tumor cell lines sensitive or resistant to doxorubicin. N-Trifluoroacetylation of the antibiotics resulted in a decrease in the cytostatic activity. The activity of the water-soluble 13-(4-methylpiperazine-l-yl)imino derivatives is close to that of the corresponding parent antibiotics.

[Derivatives of antineoplastic antibiotics of the daunorubicin series containing methylurea or nitrosomethylurea residues]. / Proizvodnye protivoopukholevykh antibiotikov riada daunorubitsina, soderzhashchie ostatok metilmocheviny ili nitrozometilmocheviny.

Derivatives of antitumour anthracycline antibiotics containing N-methylurea moiety in the carbohydrate ring were obtained by the interaction of methyl isocyanate with daunorubicin, doxorubicin, carminomycin and daunorubicin derivatives, substituted at C-13 or C-14 positions. N-Nitrosation of these compounds yielded modified anthracycline antibiotics containing the N-methyl-N-nitrosourea substituent at C-3' position. Alkaline degradation of these derivatives produced, through corresponding isocyanates cyclic 3'-N,4'-carbonylderivatives. In these anthracycline derivatives with sugar cycles conjugated with oxazoline-2-ones the predominant conformations of sugar ring has changed from 1C4 to 4C1, 2,5B, or B0,3 (shown by 1H NMR spectroscopy). It was demonstrated, both in vitro and in vivo, that introduction of methylurea or cytotoxic methylnitrosourea moieties does not potentiate antimicrobial, cytotoxic or antitumour properties of these compounds.

[Synthesis and antitumor properties of 3'-desamino-3'-dimethylformamidinorubomycin]. / Sintez i protivoopukholevye svoistva 3'-dezamino-3'-dimetilformamidinorubomitsina.

Interaction of rubomycin (daunorubicin) chlorhydrate with dimethylformamidine diethyl acetal yielded 3'-desamino-3'dimethylformamidinorubomycin chlorhydrate (DFR). Comparative antitumor activity of DFR and rubomycin was studied on mice with respect to ascitic lymphadenosis NK/Ly and Ehrlich carcinoma, hemocytoblastosis La, leukemia P-388 and two solid tumors i. e. lymphosarcoma LIO-I and sarcoma 180. The highest antitumor effect of DFR was observed in the mice with Ehrlich carcinoma and lymphadenosis NK/Ly after the drug intravenous administration for 4 times. By selectivity of the antitumor effect DFR was inferior to rubomycin with respect to lymphosarcoma LIO-I and sarcoma 180. It was shown that the antileukemic activity of DFR and rubomycin with respect to hemocytoblastosis La was practically the same. In the experiments with leukemia P-388 DFR was inferior to rubomycin.

[Synthesis and antitumor properties of 3'-desamino- dimethylformamidine doxorubicin]. / Sintez i protivoopukholevye svoistva 3'-dezamino- 3'-dimethylformamidinodoksorubitsina.

Interaction of doxorubicin hydrochloride with dimethylformamide diethylacetal yielded hydrochloride of 3'-desamino-3'-dimethylformamidine doxorubicin (DFD). It was shown that with single intravenous administration to tumor-free mice DFD was 2.5 times less toxic than the initial doxorubicin. Antitumor activity of DFD was studied with respect to 6 transplantable murine tumors: lymphosarcoma LIO-1, sarcoma 180, lymphadenosis NK-Ly, Ehrlich carcinoma, hemocytoblastosis La and leukemia P-388. Selectivity of the DFD activity against all the above tumors was shown to be high and practically equal to that of doxorubicin. DFD had the highest inhibitory effect on development of Ehrlich carcinoma and lymphosarcoma LIO-1.

[Study of toxicity and antineoplastic activity of 14-substituted derivatives of rubomycin and carminomycin]. / Izuchenie toksichnosti i protivoopukholevoi aktivnosti 14-zameshchennykh proizvodnykh rubomitsina i karminomitsina.

Toxicity and antitumor activity of five derivatives of rubomycin and carminomycin were studied in animals. The derivatives were prepared by modification of the methyl C-14 group. These were the following: 14-chlorrubomycin, 14-chlorcarminomycin, 14-salicyloyloxyrubomycin, 14-salicyloyloxycarminomycin and 14-quinaldinoyloxyrubomycin. The chemotherapeutic study revealed that, in their activity, all the compounds were inferior to the starting antibiotics. Unlike the other derivatives, 14-chlorcarminomycin induced a significant inhibition of leukemia P-388 development (the average lifespan of the mice amounted to 165 per cent as compared to the control). However, in the magnitude of its effect, the derivative was inferior to carminomycin.

[Production and antineoplastic activity of new asymmetric azines on the rubomycin base]. / Poluchenie i protivoopukholevaia aktivnost' novykh nesimmetrichnykh azinov na osnove rubomitsina.

New alkulidene hydrazones of rubomycin (daunorubicin) with the linear or branched chain of the carbon atoms were studied: rubomycin 13-(hexylidene-2")-hydrazone, rubomycin 13-(heptylidene-3")-hydrazone and rubomycin 13-(4"-methylpentylidene-2")-hydrazone. Alkylidene hydrazones of the formamidine derivatives were also studied: 13:cyclohexylidene hydrazone of 3'-desamino-3'-dimethylformamidine rubomycin and 13-(5"-oxypentyliden-2") hudrazone of 3'-desamino-3'-dimethylformamidine rubomycin. The latter two alkylidene hydrazones were modified twice. It was found that after a single intravenous administration to tumor-free mice the new substance had the same or lower toxicity as compared to that of rubomycin. Antitumor activity of the substances against lymphosarcoma LIO-I was studied comparatively with that of the initial rubomycin. It was shown that the molecule modification at C-13, as well as simultaneous modification at C-13 and the sugar amino group resulted in lowering of the antitumor activity in comparison to that of the starting rubomycin.

[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.

[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).

[Obtaining 14-oxycarminomycin and a study of its antitumor activity]. / Poluchenie 14-oksikarminomitsina i izuchenie ego protivoopukholevoi aktivnosti.

The authors obtained 14-oxykarminomycin by alkaline hydrolysis of 14-bromokarminomycin. On two-fold intravenous administration to mice with lymphosarcoma, strain L10-1, 14-oxycarminomycin showed the same toxicity as karminomycin. The preparation had the same selective antitumor activity as karminomycin.

[Synthesis and properties of carminomycinone derivatives]. / Sintez i svoistva proizvodnykh karminomitsinona.

The possibility of chemical modification of carminomycinone-aglycone (II) of carminomicin, a side product in the antibiotic production was studied. The methyl group C-14 was functionilized by introducing the bromine atom and performing a number of exchange reactions with the bromine atom. It was found that under definite conditions (1. 1 equiv. Br2in dioxane, 20 degrees, 24 hours) carminomycinone (II) was subjected to selective bromination into the side acetyl group with formation of 14-bromcarminomycinone (III). On interaction with anhydrous potassium acetate 14-bromcarminomycinone (III) yield 14-acetoxycarminomycinone (IV). In its turn the latter compound (IV) easily hydrolized to 14-oxycarminomycinone (V) in treatment with aqueous alkali or acid. 14-oxycarminomycinone (V) was also prepared with a high yield (80 per cent) by direct alkaline hydrolysis of 14-bromcarminomycinone (III) in treatment with 0.1N solution of sodium carbonate in a mixture of dioxane and water. The structure of 14-substituted derivatives of carminomycinone was proved by analytical and spectral data and confirmed by their transformation. Thus, according to the data of mass-spectrometry 14-oxycarminomycinone (V) had a molecular weight of 400 c. u. In treatment with an excess of acetic anhydride in pyridine it formed a hexa-acetyl derivative, i.e. 4, 6, 7, 9, 11, 14-hexa-acetyl-14-oxycarminomycinone (VI). The aglycones (III-V) prepared by us may serve a starting material in chemical synthesis, as well as biosynthesis of semi-synthetic preparations of the carminomycin series.

[Synthesis and antitumor properties of rubomycin 13-cyclohexylidene hydrazone]. / Sintez i protivoopukholevye svoistva 13-tsiklogeksildengidrazona rubomitsina.

Rubomycin 13-cyclohexylidene hydrazone (RCH) was synthesized by interaction of rubomycin 13-hydrazone with cyclohexane. Antitumor activity of RCH was studied in comparison to that of the initial rubomycin on mice with ascitic lymphadenosis NK/Ly, Ehrlich carcinoma, hemocytoblastosis La, leukemia P-388, solid lymphosarcoma LIO-1 and sarcoma 180. It was shown that RCH was actually equal to rubomycin by its selective activity against lymphadenosis NK/Ly and sarcoma 180 on its 4- and 2-fold intravenous administration, respectively. RCH was superior to rubomycin by its selective activity against lymphosarcoma LIO-1 and Ehrlich carcinoma and was more efficient in treatment of hemocytoblastosis La. RCH was inferior to rubomycin by its efficacy in treatment of lymphadenosis NK/Ly on its 2-fold oral administration and against leukemia P-388 on its 2-fold intravenous administration. Comparison of RCH with an analogous derivative of carminomycin showed that analogous changes in the chemical structure of rubomycin and carminomycin induced different changes in the chemotherapeutic properties of the initial substances.

[Synthesis of spin-labeled rubomycin and a study of its interaction with DNA]. / Sintez spin-mechenogo rubomitsina i izuchenie vzaimodeistviia ego s DNK.

Hydrochloride of 14-(I-oxyl-2,2,6,6-tetramethylpiperidyl-4)-acetoxyrubomycin (spin-labeled rubomycin or SL-rubomycin) was prepared by interaction of hydrochloride of 14-bromrubomycin with potassium salt of I-oxyl-2,2,6,6-tetramethylpiperidyl-4-acetic acid. Its interaction with DNA and synthetic poly A and poly U polyribonucleotides was studied. The character of the EPR spectra was indicative of DNA binding with SL-rubomycin and forming a system with a high level of regularity similar to that of liquid crystals. The results of the study of the EPR spectra correlated with the model of rubomycin intercalation between the pairs of DNA bases and were indicative of water surrounding of the nitroxyl group of SL-rubomycin bound with DNA.

[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.

[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.

[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.

[Preparation of dihydrocarminomycin and a comparison of its antitumor activity with the activity of carminomycin]. / Poluchenie digidrokarminomitsina i sravnenie ego protivoopukholevoi aktivnosti s aktivnost'iu karminomitsina

A dihydro derivative of karminomycin was prepared using chemical reduction with potassium boron hydride. When dihydrokarminomycin was administered intravenously to healthy albino mice in a single dose it practically showed the same toxicity as karminomycin. However, unlike the latter dihydrokarminomycin induced the death of the animals at later periods of time. Studies on mice with transplantable tumours showed high antitumor activity of dihydrokarminomycin against lymphosarcoma L10-1, sarcoma 180, Garding-Passy melanoma, lymphoid leukosis L-1210 and lymphocytal leukosis P-388. In treatment of the mice with leukosis L-1210 and Garding-Passy melanoma dihydrokapminomycin was much inferior by its efficiency than karminomycin.

[Antiviral action of carminomycin and some of its derivatives]. / Protivovirusnoe deistvie karminomitsina i nekotorykh ego proizvodnykh.

Carminomycin was shown to inhibit the development of both the DNA-containing variolovaccine virus and the RNA-containing grippe virus in chick embryos. Comparison of the effects of rubomycin, carminomycin, 14-oxy-carminomycin and carminomycin complex with bovine serum albumin in experiments with chick embryos showed that the inhibitory effect of carminomycin and its derivatives on the development of the grippe virus was much higher than that of rubomycin. The carminomycin derivatives proved to be much more active in this respect than the initial antibiotic. Carminomycin and its derivatives had a therapeutic effect on mice with experimental grippe pneumonia also on their oral use.