ANTHRACYCLINE COMPOUNDS AND USE THEREOF

Abstract: An anthracycline compound selected from the group consisting of anthracycline compounds represented by the following general formula [A], acid addition salts thereof, anthracycline compounds represented by the following general formula [B], and acid addition salts thereof: wherein R¹ to R⁴ have the following meanings: (i) R¹ and R² each represent a hydrogen atom or a hydroxyl group; (ii) R³ represents a hydrogen atom or a group (iii) if R³ is a hydrogen atom, R⁴ represents either one of the following groups (a) to (i): wherein R⁵ represents a hydrogen atom, an alkyl group, an alkenyl group, an alkynyl group, a fluoroalkyl group, an aryl group, an aralkyl group, or a group -O-(CH₂) n -X-(CH₂) n COOH, wherein X represents a hetero atom and n denotes an integer of 1 to 20; wherein R⁶ and R⁷ respectively represent a hydrogen atom, an alkyl group, an alkenyl group, an alkynyl group, an aryl group, an aralkyl group or an alkyl group which may have a hydroxyl group, a carboxyl group, an amino group or an ether, sulfide or amide bonding, provided that R⁶ and R⁷ will not represent a hydrogen atom at the same time; wherein R⁸ represents an alkylene group which may have a hydroxyl group, a carboxyl group or an amino group or an ether, sulfide or amide bonding; wherein R⁹ and R¹⁰ respectively represent an alkyl group, provided that R⁹ and R¹⁰ will not represent a methyl group at the same time; wherein R¹¹ represents an alkylene group except that R¹¹ is -(CH₂)₅-; wherein R¹² represents the part except the aldehyde group of a 16 membered macrolide having the aldehyde group at the side chain at the 6-­ position of an aglycone; wherein R¹³ represents an alkylene group which may include a hydroxyl group, a carboxyl group, an amino group or an ether, sulfide or amide bonding, provided that R⁴ will not represent a group (iv) if R³ is a group R⁴ represents a group wherein R¹⁴ represents a hydrogen atom or a hydroxyl group, and R¹⁵ represents a group

Description: BACKGROUND OF THE INVENTION The present invention relates to novel anthracycline compounds and further to the use of the novel anthracycline compounds, that is, the use thereof as antitumor agents. Daunomycin (US Patent No. 3,616,242) and adriamycin (US Patent No. 3,590,028) are known as anthracycline type compounds. These compounds are extensively used in clinical stages. However, they are known to have serious side effects such as cardiotoxicity, impairment of bone marrow and the like, and thus their uses remain only unsatisfactory. On the other hand, preparation of novel and miscellaneous analogous compounds by methods including fermentation, semi-synthesis, microbial transformation and the like have been attempted, and there have already been reported several compounds such as 4'-epiadriamycin (Japan Science Society Press, Tokyo/University Park Press, Baltimore, 292 - 312, 1978), 4-demethoxyadriamycin (Cancer Treat. Rep., 60: 829, 1976), and THP-adriamycin (J. Antibiotics 32: 1082, 1979). For the purpose of providing a useful anthracycline derivative, we have also proposed morpholino derivatives of 13-deoxocarminomycin, 13-deoxo-10-hydroxycarminomycin and 13-deoxo-11-deoxycarminomycin (referred to hereinafter as R20 substances) represented by the formula (I) (Japanese Patent Application Laid-Open Nos. 167696/86 and 16495/87): (1) 13-deoxocarminomycin (referred to hereinafter as R20X), R a : OH, R b : H, (2) 13-deoxo-10-hydroxycarminomycin (referred to hereinafter as R20X2), R a : OH, R b : OH, and (3) 13-deoxo-11-deoxycarminomycin (referred to hereinafter as R20Y5), R a : H, R b : H. A similar compound as mentioned unter (2), i.e. R20X2, in which the amino group has been dimethylated is known from Chem. Abstracts, 101 (1984), page 483, Ref. No. 5554x, corresponding to DD-A-205 696, as having antileukemic activity in mice. SUMMARY OF THE INVENTION The present invention relates to novel compounds corresponding to the derivatives of R20 substances represented by the above formula (I). In other words, the anthracycline compounds according to the present invention are selected from the group consisting of an anthracycline compound represented by the following general formula [A] or an acid addition salt thereof. The present invention also relates to the use of the novel compounds. The antitumor agents according to the present invention contain as an active ingredient the anthracycline compound represented by the following general formula [A] or an acid addition salt thereof: wherein: (i) R¹ represents a hydroxyl group; (ii) R² represents a hydrogen atom or a hydroxyl group; (iii) R³ represents a hydrogen atom; (iv) R⁴ represents one of the following groups (a) to (d); (a) (b) (c) (d) , wherein when R² is a hydroxyl group, R⁴ is not (d). DETAILLED DISCRIPTION OF THE INVENTION Production of the compounds of the present invention Preparation of the parent compound The compounds of the present invention are produced from the R20X and/or R20X2, which will be herinafter referred to as "the R20 substances" as primary raw materials. The R20 substances are well-known substances, and the method of preparation and the physicochemical properties thereof are as described in Japanese Patent Application Laid-Open Nos. 167696/86 and 16495/87. Further, the microorganism to be used for the preparation of the R20 substances has been deposited with Fermentation Research Institute, of the Agency of Industrial Sciences and Technology, Japan with "FERM BP945". Production of the compounds of the present invention The process for producing derivatives of the R20 substances as the leading compounds is described below. The compounds described in the paragraph (iv)-(a) can be produced by reacting the R20 substance represented by the formula (I) with N-trifluoroacethyl-4-piperidone. The solvents used for the reaction include acetonitrile, methanol, ethanol, water, chloroform, dichloromethane, carbon tetracloride or the like alone or as a mixture thereof, preferably a mixed solvent of chloroform and methanol. The reaction is generally conducted preferably in the presence of a reducing agent such as sodium borohydride or sodium cyanoborohydride. The reducing agent is used in an amount of at least 1 mole, preferably 1 - 3 moles to 1 mole of the R20 substance. N-trifluoroacetyl-4-piperidone is conveniently used in an amount of 1.5 moles or more, preferably 5 moles or more to 1 mole of the R20 substance. The reaction temperature is generally within the range from freezing point of a solvent used to 50°C, preferably around room temperature. Alternately, the compounds described in the paragraph (iv)-(a) can be produced by reacting the R20 substances with N-trifluoroacetyl-4-halopiperidine . The reaction is generally carried out in a solvent. The solvents which can be used include aprotic solvents such as N, N-dimethylformamide, tetrahydrofuran and the like. The reaction is ordinarily conducted in the presence of a base such as triethylamine or diisopropylethylamine. The base is used in an amount of at least 1.5 moles or more, preferably 2 to 5 moles per 1 mole of the R20 substance. N-trifluoroacetyl-4-halopiperidine is preferably used in an amount of 2 to 4 moles to 1 mole of the R20 substance. The reaction is preferably conducted at a temperature of 0 to 50°C, particularly around room temperature. The compound described in the paragraphs (iv)-(b), (c) and (d) can be produced by reacting the R20 substance with a compound having a dialdehyde at the terminal or a compound having a removing group such as halogen or the like at the terminal. The reaction is conducted under the same reaction conditions as those for producing the compounds described in the paragraph (iv)-(a). The compounds represented by the formula [A] can be isolated from the reaction mixture by a purification method well-known in the art of preparing gylcoside derivatives of anthracyclines. For example, the desired substance can be isolated in a pure form by extracting the reaction mixture with an organic solvent immiscible with water, preferably chloroform, methylene chloride or the like, concentrating the solvent layer, and subjecting the residue to a separation and purification procedure such as silica gel column chromatography or preparative thin-layer chromatography on a silica gel or by gel filtration chromatography on Sephadex LH20 or the like. The group of the compounds represented by the formula [A] can be converted to their acid addition salts according to a method well-known per se by treating with an inorganic acid such as hydrochloric acid, sulfuric acid, or phosphoric acid or an organic acid such as acetic acid, propionic acid, maleic acid, oleic acid, palmitic acid, citric acid, succinic acid, tartaric acid, fumaric acid, glutamic acid, pantothenic acid, or laurylsulfonic acid. Specific Examples of the Compounds Specific examples of the compounds obtained according to the present invention are as follows. 1. N-(1''-trifluoroacetyl-4''-piperidinyl)-13-deoxocarminomycin, 2. N-(1''-trifluoroacetyl-4''-piperidinyl)-13-deoxo-10-hydroxycarminomycin, 3. 3'-deamino-3'-(1''-acetyl-4''-piperazinyl)-13-deoxocarminomycin, 4. 3'-deamino-3'-(1''-acetyl-4''-piperazinyl)-13-deoxo-10-hydroxycarminomycin, 5. 3'-deamino-3'-[(2''R)-2''-acetamido-4''morpholino]-13-deoxocarminomycin, 6. 3'-deamino-3'-[(2''R)-2''-acetamido-4''-morpholino]-13-deoxo-10-hydroxycarminomycin, 7. A, B 3'-deamino-3'-(6''hydroxy-2''-methoxy-4''-perhydroxazepinyl)-13-deoxocarminomycin, wherein A and B are stereoisomers. Structure The compounds 1 to 7 have the following structures for R¹, R², R³ and R⁴ in the forumla [A]: Use of the Derivatives The derivatives of the present invention have carcinostatic activity and thus are useful as a medicament. (1) Antitumor agent The derivatives of the present invention exhibited antitumor activities with respect to leukemia of experimental animals as shown in Table 1. The data shown in Table 1 are the survival rate(%) of CDF₁ mice, to which a suspension of P388 leukemia cells have been inoculated intraperitoneally in an amount of 1 x 10⁶/mouse, and the derivative has been administered intraperitoneally (ip) or intravenously (iv) on day 1 and day 5 after inoculation, based on the survival days of the control group to which physiological saline has been administered. (2) Antitumor agent As shown above, the derivatives of the anthracycline compounds of the present invention have been confirmed to have antitumor activity relative to animal tumors, particularly malignant tumors. Accordingly, the derivatives of the present invention can be used as an antitumor agent or a tumor therapeutic agent. The derivatives of the present invention as the antitumor agent can be administered in an appropriate optional administration route or in a dosage form which will be determined by the adopted administration route. The derivatives as a medicament has generally a form which has been diluted with a pharmaceutically acceptable carrier or diluent. When the derivative of the present invention as the antitumor agent is practically administered, it may typically be injected as a solution in distilled water for injection or physiological saline. Specifically for veterinary application, the injectional methods of intraperitoneal injection, subcutaneous injection, intravascular injections such as intravenous or intra-arterial injection and local application may be used, and for the application to human subjects injectional methods including intra-vascular injections such as intravenous or intra-arterial injection or local application may be used. The dose of the derivatives of the present invention is determined to ensure that the total dose of continuous or intermittent dosage will not exceed a certain level in consideration of the results of animal tests and various situations. It is needless to say that the dose of the derivative specifically varies depending on application methods, situations of patients or subject animals such as age, body, weight, sex, sensitivity, feed, application time, drugs to be used together, patients or the conditions of the patients. Appropriate dose and administering times under a certain condition must be determined by an optimal dosage determining test by a specialist medical doctor based on the above instruction. Experimental Examples The present invention will be illustrated more fully and specifically by the following experimental examples. The present invention is not intended to be limited to these examples. Referential Example The percentage (%) means hereinafter "w/v %". Referential Example 1 (preparation of the R20 substance) (1) Preparation of mother seed The fermentation broth used is one in which the ingredients having the following contents have been dissolved in 1 liter of water and then adjusted to pH 7.2. Polypeptone 1% Molasses 1% Meat extract 1% A 100 ml portion of the above fermentation broth was placed and sterilized in a 500-ml Erlenmeyer flask, and one platinum loopfull of Actinomadura roseviolacea R20 taken from a slant was inoculated into the flask and cultured in a rotary shaker (200 rpm) at 27°C for 5 days to obtain a mother seed. (2) Cultivation The fermentation broth used was the one in which the ingredients having the following contents had been dissolved in 1 liter of water and adjusted to pH 7.4. Glucose 2.5% Soy bean powder 1.5% Dry yeast 0.2% (Precipitated) calcium carbonate 0.4% Twenty-five liters of the above-mentioned fermentation broth was placed and sterilized in a 50-liter jar-fermenter. Three vials of the above-mentioned mother seeds were inoculated into the fermentation broth, and cultured with aeration (1 vvm) and agitation (200 rpm) at 27°C for 7 days. (3) Collection of R20X After cultivation, the fermentation broth was filtered to separate the bacterial cells and the filtrate. The filtrate was adjusted to pH 2 with 1N HCl and adsorbed on a column (10 x 40 cm) of " Diaion HP20" (manufactured by Mitsubishi Chemical Industries Ltd.). After washing with distilled water and 60% methanol, elution was conducted with methanol. The eluate was concentrated. The concentrate was adjusted to pH 8.5 and extracted three times with a mixture of chloroform-methanol (9:1). The extract was concentrated, and hexane was added in an amount of 6 times that of the concentrate. After the precipitate thus produced was dried, 250 mg of the red powder was obtained. The powder was dissolved in chloroform, placed on a 4 x 40 cm column which had been equilibrated with chloroform, rinsed well with chloroform and then fractionated with chloroform-methanol (10:1). The active fraction thus obtained was concentrated under reduced pressure, developed on a TLC plate ("Silica gel 60" manufactured by Merck Co.) with a solvent system of chloroform-methanol-acetic acid-water (40:8:1:1) to collect a reddish orange fraction at an Rf value of about 0.43 by scraping. The fraction thus obtained was eluted, concentrated and then recrystallized from chloroform to obtain 110 mg of R20X. (4) Preparation of R20X2 After cultivation, the fermentation broth was filtered to separate the bacteria cells and the filtrate. The filtrate was adjusted to pH 2 with 1N HCl and adsorbed on a 10 x 40 cm column of " Diaion HP20" (manufactured by Mitsubishi Chemical Industries Ltd.). After washing with distilled water and 50% methanol, elution was conducted with methanol. The eluate was concentrated. The concentrate was adjusted to pH 8.5 and extracted three times with a mixture of chloroform-methanol (9:1). The extract was concentrated, and hexane was added in an amount of 6 times that of the concentrate. The precipitate thus produced was dried to obtain 250 mg of a red powder (R20X2 crude product). The crude product of the R20X2 (250 mg) was dissolved in chloroform, placed on a 4 x 40 cm silica gel column (250 g) which had been equilibrated with chloroform, rinsed well with chloroform and then eluted with chloroform-methanol (10:1). The fraction thus obtained was concentrated under reduced pressure, developed on a TLC plate ("Silica gel 60" manufactured by Merck Co.) with a solvent system of chloroform-methanol-aqueous ammonia (8:2:0.05) to collect an orange fraction at an Rf value of about 0.44 by scraping. The fraction thus obtained was eluted, concentrated and then recrystallized from chloroform to obtain 10 mg of R20X2. Example 1 In 30 ml of pyridine was dissolved 1 g of 4-piperidone hydrochloride, and 10 ml of trifluoroacetic anhydride was added on an ice bath. After addition, the mixture was stirred at room temperature for 15 hours, diluted with water and extracted with chloroform. The chloroform layer was washed with 0.01N aqueous NaOH, 0.01N aqueous HCl and water, dried and then concentrated to dryness to obtain 0.93 g of N-trifluoroacetyl-4-piperidone (m.p. 68°C, IR: 1730, 1720, 1690 cm⁻¹). In methanol was dissolved 50.2 mg of R20X, and 36.3 mg of N-trifluoroacetyl-4-piperidone and 12.6 mg of sodium cyanoborohydride were added to the solution. The mixture was stirred at room temperature for 4 hours, concentrated and subjected to silica gel thin-layer chromatography with a solvent system of chloroform-methanol (9:1). The colored portion was collected by scraping, eluted with a mixture of chloroform-methanol, concentrated and subjected to gel filtration on Sephadex LH20 with a solvent system of chloroform-methanol (1:1) to obtain 36.5 mg of N-(1''-trifluoroacetyl-4''-piperidinyl)-13-deoxocarminomycin. In the same manner, 153 mg of R20X2 was dissolved in methanol, and 111 mg of N-trifluoroacetyl-4-piperidone and 37.4 mg of sodium cyanoborohydride were added to the solution. After the reaction, 95.0 mg of N-(1''-trifluoroacetyl-4''-piperidinyl)-13-deoxo-10-hydroxycarminomycin was obtained by the same purification procedure as described above. Physicochemical properties of N-(1''-trifluoroacetyl-4''-piperidinyl)-13-deoxocarminomycin: Molecular formula: C₃₃H₃₇O₁₀N₂F₃ Molecular weight: 678 (FD-MS) Elementary analysis: Calculated Found C 58.40 58.32 H 5.50 5.73 N 4.13 4.01 Specific rotation: [α] 23 / D = +366° (c = 0.28, in 0.01N HCl-MeOH), Melting point: 136 - 139°C UV-Visible spectrum 235(903) 240(913) 255(699) 299(183) 293(210) 551(330) 492(378) (in alkaline methanol) 527(256) (in acidic methanol) IR spectrum: 3400, 1700, 1600 cm⁻¹ (KBr) Proton NMR spectrum δ H : 1.10(3H, t, J=7.8Hz, H-14), 1.15 - 1.40(2H, m, H-2'), 1.38(3H, d, J=6.5Hz, H-6'), 1.60 - 1.98(7H, m, H-8a, H-13, H-3'', H-5''), 2.42(1H, d, J=15Hz, H-8b), 2.53(1H, dd, J=19Hz, 2.5Hz, H-10a), 2.78 - 3.15(4H, m, H-3', H-2''a, H-6''a, H-4''), 3.23(1H, d, J=19Hz, H-10b), 3.55(1H, brs, H-4'), 3.8 - 4.35(3H, m, H-5', H-2''b, H-6''b), 5.15(1H, brs, H-7), 5.45(1H, brs, H-1'), 7.23(1H, d, J=8.0Hz, H-3), 7.65(1H, dd, J=8.0, 8.0Hz, H-2), 7.80(1H, d, J=8.0Hz, H-1) Physicochemical properties of N-(1''-trifluoroacetyl-4''-piperidinyl)-13-deoxo-10-hydroxycarminomycin: Molecular formula: C₃₃H₃₇O₁₀N₂F₃ Molecular weight: 694 (FD-MS) Elementary analysis: Calculated Found C 57.06 56.82 H 5.37 5.49 N 4.03 3.95 Specific rotation: [α] 24 / D = +466° (c = 0.5, in 0.01N HCl-MeOH), Melting point: 128 - 131°C UV-Visible spectrum 235(660) 239(654) 254(399) 297(129) 292(139) 552(225) 494(249) (in alkaline methanol) 528(164) (in acidic methanol) IR spectrum: 3500, 1700, 1600 cm⁻¹ (KBr) Proton NMR spectrum δ H : 1.12(3H, t, J=7.3Hz, H-14), 1.36(3H, d, J=6.2Hz, H-6'), 1.20 - 1.95(6H, H-13, H-2', H-3'', H-5''), 2.15(1H, dd, J=3.5, 14.8Hz, H-8a), 2.24(1H, d, J=14.8Hz, H-8b), 2.8 - 3.2(6H, H-3', H-4'', H-2''a, H-6''a), 3.56(1H, brs, H-4'), 3.80 - 4.32(6H, H-2''b, H-6''b, H-5'), 4.85(1H, s, H-10), 5.12(1H, brs, H-7), 5.45(1H, brs, H-1'), 7.30(1H, d, J=8.0Hz, H-3), 7.74(1H, dd, J=8.0, 8.0Hz, H-2), 7.82(1H, d, J=8.0Hz, H-1) Example 2 In 10 ml of pyridine was dissolved 0.5 g of diallylamine hydrochloride, and 1.0 g of acetic anhydride was added to the solution. The mixture was stirred at room temperature for 10 hours. Water was addd to the mixture and the mixture was extracted with chloroform. The chloroform layer was washed with 0.01N aqueous HCl solution, 1% aqueous sodium bicarbonate solution and finally with water, and concentrated to obtain 0.47 g of N-acetyldiallylamine [IR: 1640 cm⁻¹, ¹H-NMR δ H 2.08 (3H), 3.94(4H), 5.16(4H), 5.76(2H)]. In 150 ml of methylene chloride was dissolved N-acetyldiallylamine, and the solution was treated with ozone at -65°C for 10 hours. After 10 hours, 20 ml of dimethylsulfide was added to the mixture, and the mixture was stirred at room temperature for further 10 hours. The solvent was removed under reduced pressure, and a solution of 40 mg of R20X in 5 ml of methanol was added to the residue. Then, 10.1 mg of sodium cyanoborohydride was further added to the mixture and the mixture was stirred at room temperature for 1 hour. Water was added to the reaction mixture, and the mixture thus formed was extracted with chloroform. The chloroform layer was dried, concentrated and subjected to silica gel thin-layer chromatography with a solvent system of chloroform-methanol (10:1). The colored band at an Rf value of ca. 0.4 was scraped off, eluted with a mixed solvent of chloroform-methanol and concentrated. The concentrate was subjected to gel filtration on Sephadex LH20 with a solvent system of chloroform-methanol (1:1) and the fraction thus obtained was concentrated. The concentrate was dissolved in a small amount of chloroform, and 20 ml of hexane was added to the solution. The precipitate thus formed was collected by centrifugation and dried to obtain 12.4 mg of 3'-deamino-3'-(1''-acetyl-4''-piperazinyl)-13-deoxocarminomycin. In the same manner, 25.4 mg of 3'-deamino-3'-(1''-acetyl-4''-piperazinyl)-13-deoxo-10-hydroxycarminomycin was obtained from 110 mg of R20X2. Physicochemical properties of 3'-deamino-3'-(1''-acetyl-4''-piperazinyl)-13-deoxocarminomycin: Molecular formula: C₃₂H₃₈O₁₀N₂ Molecular weight: 610 (FD-MS) Elementary analysis: Calculated Found C 62.94 61.70 H 6.27 6.15 N 4.59 4.44 Specific rotation: [α] 22 / D = +313° (c = 0.1, in 0.01N HCl-MeOH), Melting point: 146 - 151°C (dec.) UV-Visible spectrum 235(673) 240(698) 254(390) 297(152) 292(146) 552(260) 495(244) (in alkaline methanol) 528(165) (in acidic methanol) IR spectrum: 3400, 1730, 1620 cm⁻¹ (KBr) Proton NMR spectrum: 1.08(3H, t, J=7.8Hz, H-14), 1.38(3H, d, J=6.3Hz, H-6'), 1.5 - 2.0(5H, H-8a, H-2', H-13), 2.00(3H, s, CoCh₃), 2.2 - 2.6(6H, H-3'', H-5'', H-8b, H-10a), 3.22(1H, d, J=19.0Hz, H-10b), 3.20 - 3.60(4H, H-2'', H-6''), 3.70(1H, brs, H-4'), 4.14(1H, q, J=6.3Hz, H-5'), 5.14(1H, brs, H-7), 5.48(1H, brs, H-1'), 7.24(1H, d, J=8.0Hz, H-3), 7.64(1H, dd, J=8.0, 8.0Hz, H-2), 7.80(1H, d, J=8.0, H-1). Physicochemical properties of 3'-deamino-3'-(1''-acetyl-4''-piperazynil)-13-deoxo-10-hydroxycarminomycin: Molecular formula: C₃₂H₃₈O₁₁N₂ Molecular weight: 626 (FD-MS) Elementary analysis: Calculated Found C 61.33 61.15 H 6.11 6.28 N 4.47 4.32 Specific rotation: [α] 24 / D = +440° (c = 0.13, in 0.01N HCl-MeOH), Melting point: 143 - 147°C UV-Visible spectrum 235(641) 238(598) 254(371) 297(118) 292(132) 552(210) 494(234) (in alkaline methanol) 528(154) (in acidic methanol) IR spectrum: 3400, 1730, 1600 cm⁻¹ (KBr) Proton NMR spectrum: 1.12(3H, t, J=7.2Hz, H-14), 1.40(3H, d, J=6.5Hz, H-6'), 1.73 - 1.91(4H, H-13, H-2'), 2.13(1H, dd, J=3.8, 14.2Hz, H-8a), 2.25(1H, d, J=14.2Hz, H-8b), 2.38 - 2.62(4H, H-2'', 4''), 3.3 - 3.73(4H, H-1'', H-5''), 3.92(1H, brs, H-4'), 4.10(1H, q, J=6.5Hz, H-5'), 4.90(1H, s, H-10), 5.14(1H, dd, J=1Hz, 3.8Hz, H-7), 5.51(1H, brs, H-1'), 7.32(1H, d, J=8.2Hz, H-3), 7.72(1H, dd, J=8.2, 8.2Hz, H-2), 7.87(1H, dd, J=8.2, H-1) Example 3 In 100 ml of pyridine was dissolved 5.08 g of D-(+)-xylose, and 20 ml of acetic anhydride was added to the solution on an ice bath. The mixture was stirred at room temperature for 5 hours. Water was added to the mixture and the mixture was extracted with chloroform. The chloroform layer was washed with 0.1N HC1, 1% sodium bicarbonate solution and finally with water, dehydrated and concentrated to obtain 11.32 g of the tetra-o-acetyl derivative of xylose. The product was dissolved in 100 ml of methylene chloride, and 17.28 g of 25% hydrobromide/acetic acid solution was added and stirred. After 4 hours, water was added to the mixture, and the mixture was extracted with methylene chloride. The methylene chloride layer was washed twice with 1% sodium bicarbonate solution and further with water. The organic layer was concentrated and crystallized from a solvent system of diethyl ether-hexane to obtain 8.47 g of 2,3,4-tri-O-acetyl-α-D-xylopyranosyl bromide. [α] 24 / D = +181° (c = 0.5, in chloroform), Melting point: 92°C, Proton NMR spectrum δ H 2.05 (6H, s, OAc), 2.10 (3H, s, OAc), 3.89 (1H, dd, J=11.0, 11.0Hz, H-5a), 4.05(1H, dd, J=5.0, 11.0Hz, H-5b), 4.78(1H, dd, J=1.8, 11.0Hz, H-2), 5.05(1H, ddd, J=5.0, 11.0, 11.0Hz, H-4), 5.58(1H, dd, J=11.0, 11.0Hz, H-3), 6.58(1H, d, J=1.8Hz, H-1). In acetonitrile was dissolved 3 g of 2,3,4-tri-O-acetyl-α-D-xylopyranosyl bromide, and 2.88 g of dry sodium azide was added to the solution. The mixture was stirred at 60°C for 4 hours. Water was added to the reaction and the mixture was extracted with chloroform. The chloroform layer was washed with water, dehydrated and subjected to silica gel column chromatography with a solvent system of chloroform-acetone (100:1) to obtain 1.36 g of 2,3,4-tri-O-acetyl-1-azido-1-deoxy-β-D-xylopyranose. Specific rotation: [α] D = -123° Melting point: 76°C IR spectrum: 2130, 1750 cm⁻¹ Proton NMR spectrum δ H 2.04(3H, s, OAc), 2.05(3H, s, OAc), 2.08(3H, s, OAc), 3.44(1H, dd, J=10.0, 13.2Hz, H-5ax), 4.22(1H, dd, J=5.0, 13.2Hz, H-5eq), 4.64(1H, d, J=9.0Hz, H-1), 4.88(1H, dd, J=9.0, 9.0Hz, H-2), 4.98(1H, ddd, J=5.0, 9.0, 10.0Hz, H-4), 5.20(1H, dd, J=9.0, 9.0Hz, H-3) In 20 ml of ethyl acetate was dissolved 0.73 g of 2,3,4-tri-O-acetyl-1-azido-1-deoxy-β-D-xylopyranose, and 65 mg of 10% Pd/C was added to the solution. Then the solution was stirred in a hydrogen atmosphere for 2 hours. The Pd/C was removed by filtration, and the filtrate was concentrated. To the concentrate was added 20 ml of pyridine, 3 ml of acetic anhydride and then 10 mg of 4- dimethylaminopyridine, and the mixture was stirred for 16 hours. Water was added to the reaction, and the mixture was extracted with chloroform. The chloroform layer was washed with 0.1N HCl solution, 1% sodium bicarbonate solution and finally with water, dehydrated and concentrated. The residue was subjected to silica gel column chromatography with a system of chloroform-acetone (100:1) to obtain 0.4 mg of 1-acetamido-2,3,4-tri-O-acetyl-1-deoxy-β-D-xylopyranose. Specific rotation: [α] 21 / D = +21° (c = 0.9, in methanol) Melting point: 170°C IR spectrum: 1760, 1740, 1660 cm⁻¹ (KBr) Proton NMR spectrum: 2.00(3H, s, OAc), 2.05(3H, s, OAc), 2.06(3H, s, OAc), 2.10(3H,s, NAc), 3.35(1H, dd, J=11.0, 11.0Hz, H-5ax), 4.06(1H, dd, J=5.5, 11.0Hz, H-5eq), 4.87(1H, dd, J=10.0, 10.0Hz, H-2), 4.98(1H, ddd, J=5.5, 10.0, 11.0Hz, H-4), 5.15(1H, dd, J=10.0, 10.0Hz, H-3), 5.30(1H, dd, J=10.0, 10.0Hz, H-2), 6.39(1H, d, J=10.0, NH). In 10 ml of methanol was dissolved 380 mg of 1-acetamide-2,3,4-tri-O-acetyl-1-deoxy-β-D-xylopyranose, and 3 mg of sodium methoxide was added to the solution. The mixture was stirred at room temperature for 14 hours, then neutralized with 1N HCl, and 10 ml of water was added to the mixture. Further, 780 mg of sodium periodate was added, and the mixture was stirred for 3 hours. The reaction mixture was filtered, and the filtrate was concentrated. Methanol was added to the residue, and the resulting precipitate was separated by filtration. The filtrate was concentrated to afford 290 mg of 3-acetamido-2,6-dihydroxydioxane. In methanol was dissolved 50.2 mg of R20X, and 145 mg of 3-acetamido-2,6-dihydroxydioxane and 13.8 mg of sodium cyanoborohydride were sequentially added. The mixture was stirred at room temperature for 1 hour, then concentrated and subjected to silica gel thin-layer chromatography. The colored portion was scraped off, eluted with a mixed solvent of chloroform-methanol, concentrated and subjected to gel filtration on Sephadex LH20 with a solvent system of chloroform-methanol (1:1) to obtain 27.9 mg of 3'-deamino-3'-[(2''R)-2'-acetamido-4''-morpholino]-13-deoxocarminomycin. In the same manner, 38.7 mg of 3'-deamino-3'-[(2''R)-2''-acetamido-4''-morpholino]-13-deoxo-10-hydroxycarminomycin was obtained from 110 mg of R20X2 and 145 mg of 3-acetamido-2,6-dihydroxydioxane. Physicochemical properties of 3'-deamino-3'-[(2''R)-2''-acetamido-4''-morpholino]-13-deoxocarminomycin: Molecular formula: C₃₂H₃₈O₁₁N₂ Molecular weight: 626 (FD-MS) Elementary analysis: Calculated Found C 61.33 61.09 H 6.11 6.23 N 4.47 4.38 Specific rotation: [α] 24 / D = +326° (c = 0.2, in 0.01N HCl-MeOH), Melting point: 158 - 161°C (dec.) UV-Visible spectrum 235(626) 240(680) 255(487) 297(130) 293(140) 554(232) 492(253) (in alkaline methanol) 528(170) (in acidic methanol) IR spectrum: 3450, 1670, 1600 cm⁻¹ (KBr) Proton NMR spectrum: (in deuterochloroform) 1.09(3H, t, J=7.4Hz, H-14), 1.38(3H, d, J=6.2Hz, H-6'), 1.65 - 1.90(4H, H-13, H-2'), 2.00(3H, s, NAc), 2.1 - 2.90(7H, H-8a, 8b, 10a, H-3'', 5''), 3.28(1H, d, J=19.0Hz, H-10b), 3.68(1H, m, H-6''a), 3.70(1H, brs, H-4'), 3.83(1H, m, H-6''b), 4.15(1H, q, J=6.2Hz, H-5'), 5.22(1H, brs, H-7), 5.32(1H, ddd, J=2.5, 8.2, 8.2Hz, H-2''), 5.53(1H, brs, H-1'), 6.23(1H, d, J=8.2Hz, -NH-), 7.31(1H, d, J=8.3Hz, H-3), 7.71(1H, dd, J=8.3, 8.3Hz, H-2), 7.88(1H, d, J=8.3Hz, H-1). Physicochemical properties of 3'-deamino-3'-[(2''R)-2''-acetamido-4''-morpholino]-13-deoxo-10-hydroxycarminomycin: Molecular formula: C₃₂H₃₈O₁₂N₂ Molecular weight: 642 (FD-MS) Elementary analysis: Calculated Found C 59.81 59.63 H 5.96 6.08 N 4.36 4.24 Specific rotation: [α] 24 / D = +347° (c = 0.2, in 0.01N HCl-MeOH), Melting point: 170 - 174°C (dec.) UV-Visible spectrum 235(605) 240(562) 255(352) 297(113) 292(123) 554(184) 494(224) (in alkaline methanol) 528(148) (in acidic methanol) IR spectrum: 3450, 1660, 1600 cm⁻¹ (KBr) Proton NMR spectrum: (in deuterochloroform) 1.12(3H, t, J=7.3Hz, H-14), 1.37(3H, d, J=6.2Hz, H-6'), 1.70 - 1.90(4H, H-2', H-13), 1.98(3H, s, NAc), 2.15(1H, dd, J=4.5, 14.8Hz, H-8a), 2.22(1H, d, J=14.8Hz, H-8b), 2.25 - 2.70(4H, H-3'', H-5''), 3.65(1H, m, H-6''a), 3.73(1H, brs, H-4'), 3.80(1H, m, H-6'b), 4.09(1H, q, J=6.2Hz, H-5'), 4.85(1H, s, H-10), 5.12(1H, brs, H-7), 5.30(1H, m, H-2'), 5.49(1H, brs, H-1'), 7.29(1H, d, J=8.0Hz, H-3), 7.70(1H, dd, J=8.0, 8.0Hz), 7.81(1H, d, J=8.0Hz). Example 4 In 20 ml of methanol was dissolved 2.65 g of D-xylose, and a catalytic amount of p-toluenesulfonic acid was added. The mixture was stirred at 40°C for 3 hours, neutralized with a cation exchange resin IRA-410 and concentrated to obtain 3.09 g of methyl-D-xyloside. In 50 ml of water was dissolved 3.09 g of methyl-D-xyloside, 10.0 of sodium periodate was added, and the mixture was stirred at room temperature for 4 hours. The aqueous layer was concentrated, and a large amount of methanol was added to the concentrate. The precipitate thus formed was filtered and the filtrate was concentrated and subjected to silica gel column chromatography with a solvent system of chloroform-methanol (1:1) to obtain a compound represented by the formula (III): In methanol was dissolved 101.1 mg of the compound, and 67.3 mg of R20X and then 84.7 mg of sodium cyanoborohydride were added to the solution. The mixture was stirred at room temperature for 40 minutes, concentrated and subjected to silica gel thin-layer chromatography with a solvent system of chloroform-methanol (10:1). A scraped portion was eluted with a mixed solvent of chloroform-methanol and concentrated. The concentrate was further subjected to silica gel thin-layer chromatography with a solvent system of chloroform-methanol (10:1). A scraped colored fraction at an Rf value of 0.6 produced 22.7 mg of 3'-deamino-3'-(6''-hydroxy-2''-methoxy-4''-perhydroxazepinyl)-13-deoxocarminomycin A. Colored fraction at the Rf value of 0.55 produced 24.7 mg of 3'-deamino-3'-(6''-hydroxy-2''-methoxy-4''-perhydroxazepinyl)-13-deoxocarminomycin B. A and B are isomers at the 2''-position. Physicochemical properties of 3'-deamino-3'-(6''-hydroxy-2''-methoxy-4''-perhydrooxazepinyl)-13-deoxocarminomycin A, B: Molecular formula: A, B C₃₂H₃₉O₁₂N Molecular weight: A: 629 (FD-MS) B: 629 (FD-MS) Specific rotation: A: [α] 22 / D = +247° (c = 0.12, in 0.01N HCl-MeOH), Specific rotation: B: [α] 22 / D = +355° (c = 0.1, in 0.01N HCl-MeOH), Melting point: A: 137 - 140°C B: 139 - 143°C IR spectrum: A: 3400, 1600 cm⁻¹ (KBr) B: 3400, 1600 cm⁻¹ (KBr) Proton NMR spectrum: (in deuterochloroform) δ H A: 1.08(3H, t, 3=7.9, H-14), 1.39(3H, d, J=6.5Hz, H-6'), 1.63 - 1.86(5H, H-2', H-8a, H-13), 1.92(1H, dd, J=7.4, 10.9Hz, H-3''a), 2.08(1H, dd, J=11.0, 11.0Hz, H-5''a), 2.40(1H, d, J=15.2Hz, H-8b), 2.43(1H, m, H-3'), 2.55(1H, d, J=18.9Hz, H-10a), 2.66(1H, d, J=11.0Hz, H-5''b), 3.02(1H, d, J=7.4Hz, H-3''b), 3.24(1H, d, J=18.9Hz, H-10b), 3.49(3H, s, OMe), 3.60(1H, m, H-6''), 3.65 - 3.72(3H, H-7'', H-4'), 4.14(1H, q, H-5'), 4.57(1H, d, J=7.1Hz, H-2''), 5.19(1H, brs, H-7), 5.50(1H, brs, H-1'), 7.28(1H, d, J=8.0Hz, H-3), 7.70(1H, dd, J=8.0, 8.0Hz, H-2), 7.84(1H, d, J=8.0Hz, H-1). B: 1.08(3H, t, J=7.3Hz, H-14), 1.39(3H, d, J=6.7Hz, H-6'), 1.60 - 1.90(5H, H-13, H-8, H-2'), 2.06(1H, dd, J=11.0, 11.0Hz, H-5''a), 2.13(1H, dd, J=1.8, 11.6Hz, H-3''a), 2.48(2H, H-8a, H-3'), 2.56(1H, d, J=18.9Hz, H-10a), 2.77(1H, d, J=10.9Hz, H-5''b), 3.05(1H, d, J=11.6Hz, H-3''b), 3.25(1H, d, J=18.9Hz, H-10b), 3.35(3H, s, OMe), 3.52(1H, dd, J=4.9, 11.6Hz, H-7a), 3.61(1H, dd, J=3.6, 11.6Hz, H-7b), 3.68(1H, brs, H-4'), 3.98(1H, m, H-6''), 4.12(1H, q, J=6.7Hz, H-5'), 4.66(1H, s, H-2''), 5.09(1H, brs, H-7), 5.50(1H, brs, H-1'), 7.30(1H, d, J=7.9Hz, H-3), 7.71(1H, dd, J=7.9, 7.9Hz, H-2), 7.85(1H, d, J=7.9Hz, H-1)

Claims: An anthracycline compound selected from the group consisting of anthracycline compounds represented by the following general formula [A] and acid addition salts thereof: wherein R¹ to R⁴ have the following meanings: (i) R¹ represents a hydroxyl group; (ii) R² represents a hydrogen atom or a hydroxyl group; (iii) R³ represents a hydrogen atom; (iv) R⁴ represents one of the following groups (a) to (d); (a) (b) (c) (d) , wherein when R² is a hydroxyl group, R⁴ is not (d). N-(1''-trifluoroacetyl-4''-piperidinyl)-13-deoxocarminomycin. N-(1''-trifluoroacetyl-4''-piperidinyl)-13-deoxo-10-hydroxycarminomycin. 3'-deamino-3'-(1''-acetyl-4''-piperazinyl)-13-deoxocarminomycin. 3'-deamino-3'-(1''-acetyl-4''-piperazinyl)-13-deoxo-10-hydroxycarminomycin. 3'-deamino-3'-[(2''R)-2''-acetamido-4''-morpholino]-13-deoxo-carminomycin. 3'-deamino-3'-[(2''R)-2''-acetamido-4''-morpholino]-13-deoxo-10-hydroxycarminomycin. A, B 3'-deamino-3'-(6''-hydroxy-2''-methoxy-4''-perhydroxazepinyl)-13-deoxocarminomycin, wherein A and B are stereo isomers to each other. An antitumor agent containing as an active ingredient an anthracycline compound represented by the general formula [A] of claim 1 or an acid addition salt thereof. An antitumor agent according to Claim 9, wherein the anthracycline compound is N-(1''-trifluoroacetyl-4''-piperidinyl)-13-deoxo-carminomycin. An antitumor agent according to Claim 9, wherein the anthracycline compound is N-(1''-trifluoroacetyl-4''-piperidinyl)-13-deoxo-10-hydroxy-carminomycin. An antitumor agent according to Claim 9, wherein the anthracycline compound is 3'-deamino-3'-(1''-acetyl-4''-piperazinyl)-13-deoxocarminomycin. An antitumor agent according to Claim 9, wherein the anthracycline compound is 3'-deamino-3'-(1''-acetyl-4''-piperazinyl)-13-deoxo-10-hydroxycarminomycin. An antitumor agent according to Claim 9, wherein the anthracycline compound is 3'-deamino-3'-[(2''R)-2''-acetamido-4''-morpholino]-13-deoxocarminomycin. An antitumor agent according to Claim 9, wherein the anthracycline compound is 3'-deamino-3'-[(2''R)-2''-acetamido-4''-morpholino]-13-deoxo-10-hydroxycarminomycin. An antitumor agent according to Claim 9, wherein the anthracycline compound is 3'-deamino-3'-(6''-hydroxy-2''-methoxy-4''-perhydroxazepinyl)-13-deoxocarmino-mycin, A or B, wherein A and B are stereo isomers to each other.

Inventors: KAWAI, HIROYUKI KIRIN BEER KABUSHIKI KAISHA IYAKU, NAKAJIMA, SHOHACHI KIRIN BEER KABUSHIKI K. IYAKU, TAKEUCHI, TOMIO
Applicants: KIRIN BEER KABUSHIKI KAISHA
Application year: 1988
Main classification: C07H15
IPC codes: A61K31, C07H15, A61P35