JP2579678B2 - Anthracycline derivative and method for producing the same - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to novel anthracycline derivatives, more specifically And a salt thereof, a method for producing the same, and a method for producing adriamycin or a salt thereof from the compound of the above formula (I).

Adriamycin is a substance useful as an antibacterial agent and an antitumor agent having excellent antibacterial and antitumor effects,
Various synthetic methods are known. For example, Japanese Patent Publication No. 47-4659
No. 7 (U.S. Pat. No. 3,803,124) discloses that 14-halodaunonomycin obtained by reacting daunomycin with a halogen is reacted with an alkali metal acetate in a polar solvent, and the resulting 14-acetoxydaunomycin is alkalized. A method for producing adriamycin by hydrolysis under conditions is disclosed.

However, the method of hydrolyzing 14-acetoxydaunomycin under alkaline conditions has a serious drawback that adriamycin is decomposed and its yield is reduced because adriamycin produced is unstable to alkali.

Thus, the present inventors have conducted intensive studies on a method for producing adriamycin that does not have such disadvantages.
Novel 14-trifluoroacetyloxydaunomycin of the above formula (I) obtained by reacting halodaunonomycin with a metal salt of trifluoroacetic acid can be hydrolyzed under acidic conditions, and adriamycin can be produced in high yield. It has been found that the present invention can be manufactured at a high rate, and the present invention has been completed.

Thus, the present invention provides 14-trifluoroacetyloxydaunomycin represented by the formula (I) and a salt thereof as a novel substance.

Examples of the salt of the compound of the formula (I) include inorganic acid salts such as hydrochloride, hydrobromide and hydrogen iodide, and organic acids such as acetate, trifluoromethanesulfonate and trifluoroacetate. Salts.

The compound of the formula (I) has the following formula: In the formula, X is a halogen atom, and it can be produced by reacting 14-halodaunomycin or a salt thereof represented by the following formula with a metal trifluoroacetate.

The reaction of a compound of formula (II) or a salt thereof (eg, hydrochloride, hydrobromide, etc.) with a metal trifluoroacetate comprises
Usually, in a polar organic solvent, for example, dimethylformamide, dimethylsulfoxide, acetone, acetonitrile, methanol or the like or an aqueous mixed solvent thereof with water, a temperature in the range of about 0 ° C to about 70 ° C, preferably It can be performed at a temperature in the range of about room temperature to about 60 ° C.

The amount of the trifluoroacetate to be used with respect to the compound of the formula (II) or a salt thereof is not particularly limited.
It is appropriate to use it in an amount within the range of 1 to 15 mol, preferably 5 to 10 mol, per 1 mol of the compound of I) or a salt thereof.

The compound of the formula (II) used as a starting material in the above reaction is a compound known per se, and various methods known per se, for example, JP-B-47-46597 (US Pat. No. 3,803,124) ), JP-B-54-26529 (U.S. Pat. No. 4,225,589), and JP-B-57-135.
No. 58, Japanese Patent Publication No. 62-59719 (U.S. Pat.
No. 64) can easily be produced.

The metal trifluoroacetate to be reacted with the compound of the above formula (II) is most preferably a sodium salt or potassium salt of trifluoroacetic acid. In addition, silver salts, copper salts, tin salts, lithium salts and the like can be used. Metal salts can also be used.

The compound of formula (I) or a salt thereof obtained can be obtained by a method known per se, for example, by chromatography, crystallization,
It can be separated and purified from the reaction mixture by means such as precipitation, or can be directly subjected to the hydrolysis step described below without separation and purification.

The thus-obtained compound of the formula (I) or a salt thereof can be converted into adriamycin or a salt thereof in a high yield by hydrolysis under acidic conditions.

Hydrolysis of the compound of formula (I) or a salt thereof can be accomplished by isolating the compound of formula (I) in the isolated or reaction mixture from an aqueous solution such as water, an aqueous buffer solution or a mixture of water and a water-miscible organic solvent. It can be performed by maintaining the temperature in the ume body at a temperature of about 10 ° C. to about 70 ° C., more preferably at a temperature of about room temperature to about 60 ° C.

It is desirable to maintain the pH of the aqueous medicament during hydrolysis generally at acidic conditions in the range of 2 to 6, preferably 3 to 5.
Since the aqueous solution of the compound of the formula (I) or a salt thereof generally exhibits an acidity within the range of pH 3 to 5 depending on the kind of the salt, it is not necessary to adjust the pH during hydrolysis. If necessary, a pH adjuster such as sodium hydroxide, potassium hydroxide, sodium hydrogen carbonate, sodium carbonate, sodium phosphate or the like may be added, or a buffered aqueous solution may be used to adjust the pH of the aqueous solution to the above-mentioned range. It can also be adjusted to a range.

The hydrolysis can be usually completed in about 0.5 to 2 hours under such conditions, and the separation and purification of adriamycin or a salt thereof from the reaction mixture after the hydrolysis can be carried out by a method known per se, for example, acid transduction. , Crystallization, column chromatography and the like.

The resulting adriamycin can be converted to a salt by treatment with a suitable pharmaceutically acceptable acid, such as hydrochloric acid, hydrobromic acid, phosphoric acid, citric acid, and the like.

 Next, the present invention will be described more specifically with reference to examples.

Example 1 Japanese Patent Publication No. 56-156300 in a cutlet container (U.S. Pat.
14-bromodaunomycin hydrochloride (0.5 g, 0.778 m) prepared by the method described in Example 2 of U.S. Pat. No. 4,360,664).
mol) and sodium trifluoroacetate (1.0 g, 7.35 mmo)
l) into an eggplant flask, add 50 ml of acetone and add
For 15 hours. The formation of the reaction product (14-trifluoroacetyloxydaunomycin hydrochloride) was determined by HPLC and
After confirming by NMR spectrum, 50 m of water is contained in the reaction mixture.
l was added and the acetone was concentrated under reduced pressure. 50 ml of chloroform was added to the concentrated solution to extract the remaining acetone.

The aqueous layer was neutralized to pH 8.0 with a 5% aqueous sodium hydrogen carbonate solution, and extracted with 100 ml of chloroform-methanol (4: 1) and 50 ml of (3: 2). The chloroform extract was washed with 50 ml of a saturated saline solution, concentrated under reduced pressure, dissolved in 5 ml of methanol, added with a calculated amount of 1N HCl / methanol, and further added with isopropyl ether to crystallize adriamycin hydrochloride. The crystals are filtered, washed with isopropyl ether, and dried in a vacuum at room temperature overnight to give adriamycin hydrochloride.
0.3 g (0.517 mmol, HPLC relative purity: 91.1%, yield from 14-bromodaunomycin hydrochloride: 66.5%) was obtained.

The physicochemical properties of the intermediate 14-trifluoroacetyloxydaunomycin hydrochloride and the above adriamycin hydrochloride are as follows.

14-trifluoroacetyloxydaunomycin hydrochloride HPLC analysis (Conditions) HPLC: HPLC LC-4A manufactured by Shimadzu Corporation Column: YMC A-312 (ODS type) Mobile phase: CH ₃ CN / 0.05M NH ₄ formate buffer (pH 4. 0) Flow rate: 1.0 ml / min. Wavelength: 254 nm (Result) Elution time 14-bromodaunomycin 8.46 minutes Adriamycin 3.41 minutes 14-trifluoroacetyloxydaunomycin 22.00 minutes NMR spectrum (DMSO d ₆ ) δppm: 4.0 (3H, _{s, OCH 3) 4.9 (1H} , bs, H-7) 5.3 (1H, bs, H-1 ') 5.6 (2H, s, H-14) adriamycin hydrochloride mp: 204 - 206 ° C. (decomposition) ▲ [ α] ²⁴ _D ▼: +312.3 ゜ (c = 0.0988, CH ₃ OH) 234 nm (823.9), 252.3nm ( 542.5) 289.3nm (183.2), 476.7nm (266.2) 494.7nm (262.7), 529.4nm (137.1) IR ^{spectrum: νcm -1 (KBr) 3370 (} -OH, NH 2) , 1725 (C = O, ketone), 1610, 15
80 (C = O, quinone) NMR spectrum: δ ppm (DMSO-d ₆ / TMS) 1.16 (3H, d, J = 6 Hz, H-6 ′) 1.67 to 1.99 (2H, m, H-2 ′) 1.99 − 2.21 (2H, m, H- 8) 2.80~3.00 (2H, m, H-10) 3.55~3.70 (1H, m, H-4 ') 3.99 (3H, s, -OCH 3) 4.08~4.17 (1H , m, H-5 ') 4.60 (2H, s, H-14) 4.89 (1H, bs, H-7) 5.28 (1H, m, H-1') 5.50 (1H, s, 9-OH) 7.52 -7.70 (2H, m, H- 2, H-3) 7.80 (1H, d, J = 3Hz, H-1) 7.92-8.12 (2H, m, NH 2) 13.25 (1H, bs, 6-OH) 14.08 (1H, s, 11-OH) Example 2 14-bromodaunomycin hydrochloride (0.5 g, 0.778 mmol)
And sodium trifluoroacetate (1.0 g, 7.35 mmol) were placed in an eggplant-shaped flask, and acetone-water (1: 1) (20 ml) was added thereto.
Stirred at C for 48 hours. 30 ml of chloroform in the reaction mixture
Was added thereto, and acetone was extracted. The aqueous layer was neutralized to pH 8.0 with a 5% aqueous sodium hydrogen carbonate solution, and then extracted with 100 ml of chloroform-methanol (4: 1) and 50 ml of (3: 2).

The chloroform extract was washed with 50 ml of saturated saline and dried over anhydrous sodium sulfate. After removing sodium sulfate, concentrate under reduced pressure and dissolve in 5 ml of methanol.
N HCl / methanol was added and further isopropyl ether was added to crystallize adlimycin hydrochloride. The product was filtered, washed with isopropyl ether and dried under vacuum at room temperature overnight, and 0.3 g of adlimycin hydrochloride (0.517 mmol, H
PLC relative purity: 94.5%, yield from 14-bromodaunomycin hydrochloride: 66.5%).

Example 3 14-bromodaunomycin hydrochloride (0.5 g, 0.778 mmol)
And sodium trifluoroacetate (1.0 g, 7.35 mmol) were placed in an eggplant flask, 10 ml of DMF was added, and the mixture was stirred at 30 ° C. for 18 hours. 5 ml of water was added to the reaction solution, and the mixture was further stirred for 1 hour to produce adrimycin.

30 ml of water was added to the reaction mixture, and DMF was extracted with 80 ml and 40 ml of chloroform: methanol (3: 1).
After neutralization to pH 8.0 with a 10% aqueous sodium bicarbonate solution, adlimycin hydrochloride was extracted with 100 ml of chloroform: methanol (4: 1) and 40 ml of (3: 1). The chloroform extract was washed with 50 ml of saturated saline and dried over anhydrous sodium sulfate. After removing sodium sulfate, the solution was concentrated under reduced pressure, dissolved in 5 ml of methanol, added with a calculated amount of 1N HCl / methanol, and further added with isopropyl ether to crystallize adriamycin hydrochloride. The product was filtered, washed with isopropyl ether, and dried in vacuo at room temperature overnight. 0.31 g (0.534 mmol) of adriamycin hydrochloride, HPLC relative purity: 95.
3%, yield from 14-bromodaunomycin hydrochloride: 68.7
%).

Comparative Example 1 Production of adriamycin hydrochloride was attempted according to Example 1 of US Pat. No. 3,803,124.

14-bromodaunomycin was prepared as described in Example 1 of U.S. Pat. No. 3,803,124, but the yield was 18.3%.
Therefore, 70 ml of distilled water was added to a solution of 14-bromodaunomycin hydrochloride (1.0 g, 1.555 mmol) in 150 ml of methanol prepared by the method described in Example 2 of U.S. Pat. No. 4,360,664. 0.1N-NaOH at 20 ° C under nitrogen atmosphere
The mixture was added to pH 10.3 and hydrolyzed at the same pH for 20 minutes. 130 ml of distilled water was added to the reaction solution, and extracted five times with chloroform. The organic layer was washed with saturated saline and dried over anhydrous sodium sulfate. The solution is concentrated, and 0.6N-HCl (CH ₃
In OH, f = 0.967) 1.86 ml was added, and ether 70 ml was further added.
Was added to precipitate adriamycin hydrochloride. The precipitate was filtered off, and the filtrate was washed with 60 ml of ether and dried to give 0.6766 g of adriamycin hydrochloride (HPLC relative purity: 58
%) Obtained.

The resulting adriamycin hydrochloride was converted to a propanol-methanol system, a chloroform-methanol system or methylene chloride-as described in Example 1 of the above-mentioned U.S. Patent.
Crystallization was attempted from a methanol-based solvent, but due to low purity, crystallization was not possible, and the purity of the obtained adriamycin hydrochloride could not be increased.

Comparative Example 2 As in Comparative Example 1, when Example 1 of U.S. Pat. No. 3,803,124 was replicated as it was, high-purity adriamycin hydrochloride could not be obtained.
Production of adriamycin hydrochloride was attempted by slightly changing the operating conditions as follows.

14-bromodaunomycin was prepared as described in Example 1 of U.S. Pat. No. 3,803,124, but the yield was 18.3%.
Therefore, 14-bromodaunomycin (1.0 g, 1.555 mmol) prepared according to the method described in Example 2 of U.S. Pat. No. 4,360,664 was dissolved in 200 ml of distilled water. Under a nitrogen atmosphere, 0.1N-NaOH was added at 20 ° C. until the pH reached 10.3, and the mixture was hydrolyzed at the same pH for 20 minutes. 150m of methanol in the reaction solution
l was added and extracted five times with chloroform. The organic layer was washed with saturated saline and dried over anhydrous sodium sulfate. After removing the solvent, the residue was dissolved in chloroform: methanol (10: 1), and 0.6N-HCl (in CH ₃ OH, f = 0.96)
7) 3.01 ml was added, and the mixture was concentrated and allowed to stand at low temperature for precipitation.
The precipitate was filtered off, washed with chloroform, and dried to give 0.2152 g of adriamycin hydrochloride (HPLC relative purity: 94.1%,
Yield: 23.9%).

Comparative Example 3 According to the method described in Example 2 of U.S. Pat. No. 3,803,124, production of adriamycin hydrochloride via 14-acetoxydaunomycin was attempted.

14-bromodaunomycin was prepared as described in Example 1 of U.S. Pat. No. 3,803,124, but the yield was 18.3%.
Therefore, 0.6 g (0.933 mmol) of 14-bromodaunomycin hydrochloride prepared according to the method described in Example 2 of U.S. Pat. No. 4,360,664 was suspended in 200 mmol of anhydrous acetone, and 1.8 g of molten potassium acetate was added. And refluxed for 45 minutes. After filtering off insolubles, the filtrate was concentrated to dryness under reduced pressure. After the residue was dissolved in chloroform, ether was added, and the precipitated precipitate was filtered to obtain 0.429 g of a solid. This was purified by a silica gel column (Merck Art7734, methylene chloride: methanol: water = 100: 20: 2) to obtain 0.247 g of 14-acetoxydaunomycin.

0.1 g of 14-acetoxydaunomycin was dissolved in 30 ml of acetone: methanol (2: 1), 10 ml of a 5% aqueous sodium hydrogen carbonate solution was added, and the mixture was stirred for 3 hours. The reaction solution was diluted with water and extracted with chloroform. The extract was washed with a saturated aqueous solution of NaCl, dried over anhydrous Na ₂ SO ₄ , concentrated under reduced pressure, added with 0.6N methanolic hydrochloric acid, and further added with 3 times the amount of ether.
The precipitated precipitate was filtered and adriamycin hydrochloride 57.4 mg
(HPLC relative purity: 72.4%, total yield from 14-bromodaunomycin hydrochloride: 18.9%).

 ──────────────────────────────────────────────────続 き Continued on the front page (72) Inventor Rokuro Okamoto 5437 Fujisawa, Fujisawa City, Kanagawa Prefecture

Claims (3)

(57) [Claims] (1) Expression 14-trifluoroacetyloxydaunomycin and salts thereof represented by the formula: (2) Wherein X is a halogen atom, and 14-halodaunonomycin represented by the following formula or a salt thereof is reacted with a metal salt of trifluoroacetic acid. A method for producing 14-trifluoroacetyloxydaunomycin or a salt thereof represented by the formula: 3. The expression 14. A method for producing adriamycin or a salt thereof, comprising hydrolyzing 14-trifluoroacetyloxydaunomycin or a salt thereof under acidic conditions and, if necessary, converting the product to a salt.