CS227686B2 - A method for producing an anti-tumor active anthracycline glycoside compound - Google Patents

Abstract

The present invention relates to a process for the production of an antitumor active anthracycline glycoside compound of general formula I

Description

The present invention relates to a process for the production of an antitumor active anthracycline glycoside compound of general formula I

where .

R represents a hydrogen atom or a hydroxy group, which consists in the fact that 4-deoxy-4-iodo-N-trifluoroacetyldaunorubicin of the general formula II

dissolved in acetone is subjected to alkaline hydrolysis with 0.1 N aqueous sodium hydroxide solution carried out under mild conditions for 3 hours at 0 °C to obtain the free glycoside base of the general formula I, where R represents hydrogen, which is isolated as hydrochloride by treatment with methanolic hydrochloric acid solution and optionally reacted, dissolved in a mixture of dioxane and methanol, with a solution of bromine in chloroform to form the 14-bromo derivative, from which, after 84 hours, treatment with aqueous sodium formate solution at room temperature yields the glycoside compound of the general formula I, where R represents a hydroxy group, which is then isolated as hydrochloride.

Both new glycosides, i.e. 4-deoxy-4-iododaunorubicin and 4-deoxy-4-iodoxyrubicin, which can be produced by the method according to the invention have excellent anti-cancer activity.

The starting material for their production is - as already mentioned - 4*-deoxy-4-iodo-N-trifluoroacetyldaunorubicin. This compound is described in the owner's published European Patent Application No. 81 10 7618.1 of 24 September 1981,

The novel anthracycline glycosides of the present invention exhibit anticancer activity. In particular, 4*-deoxy-4-iododoxorubicin (a compound of the general formula I, wherein R represents a hydroxy group) has a marked effect in mouse cancer experiments.

Pharmaceutical compositions containing a therapeutically effective amount of an anthracycline glycoside of formula I are administered for therapeutic purposes in admixture with a pharmaceutically acceptable diluent or carrier and are used to treat certain mammalian malignancies by administering therapeutically effective amounts of these compounds to the affected patient.

The invention will be further described in more detail using production examples and biological data.

Example

4'-Deoxy-4-iododaunorubicin (1}R = Η) (XOO-0138)

A solution of 2.0 g of 4-iodo-N-trifluoroacetyldaunorubicin in 10 ml of acetone was treated with 80 ml of 0.1 N aqueous sodium hydroxide solution. After standing for 3 hours at 0°C, the solution was adjusted to pH 4.5 with 0.1 N aqueous hydrochloric acid and extracted with chloroform to remove the aglycones.

The aqueous solution is then adjusted to pH 8.6 and extracted with chloroform.

The combined extracts were dried over anhydrous sodium sulfate, evaporated to a small volume and acidified to pH 4.5 with 0.1 M methanolic hydrochloric acid to crystallize the title compound as its hydrochloride. Yield 1.5 g, mp 157°C (dec), mass spectrum 637 (M ⁺ ).

Chromatography on thin silica gel ^F 254 plates ( ^Merck ) using a mixture of chloroform, methanol, water and acetic acid in a volume ratio of 80:20:7:3 gives Rf = 0.72.

Example 2

4*-deoxy-4iodoxorubicin (I; R = OH) (XOO-0163)

To 1.26 g (1.87 mmol) of 4*-deoxy-4-iodo daunorubicin hydrochloride dissolved in 14 ml of methanol and 36 ml of dioxane was added 1 ml of ethyl orthoformate. The solution was treated at 10°C with 1.96 mmol of bromine in 3 ml of chloroform and 1.17 mmol of hydrogen chloride in methanol.

After one hour, the solution is poured onto a mixture of 100 ml of petroleum ether and 200 ml of diethyl ether. The precipitate formed is filtered off, dissolved in 150 ml of dioxane and 150 ml of 0.25 N hydrobromic acid and kept at 25 °C overnight. 11 g of sodium formate in 110 ml of water are added to the solution. After 24 hours, the solution is evaporated in vacuo and the residue is dissolved in a mixture of 200 ml of chloroform and methanol, both solvents being in a volume ratio of 13:6. The organic phase is washed with 5% aqueous sodium bicarbonate solution and water. After evaporation of the solvent, the residue is treated with an equivalent amount of hydrogen chloride in methanol. 0.65 g of ε^-deoxy-ε^-iododoxorubicin hydrochloride is obtained, which has a melting point of 230 °C (decomposition); mass spectrum 653 (M ⁺ ) and according to thin layer chromatography (silica gel plates ^254 ( ^Merok ) ^using a mixture of chloroform, methanol, water and acetic acid in a volume ratio of 80:20:7:3, R ^. = 0.5.

Biological activity of compounds XOO-0138 and XOO-0163:

The compounds were tested against daunorubicin (DNR) and doxorubicin (DX) in vitro against HeLa cells. The data presented in Table I show that compound XOO-0138 is less cytotoxic than DNR, while compound XOO-0.63 is more cytotoxic than DX. The anti-tumor activity was evaluated in vivo in mice suffering from P 388 ascitic leukemia. The results are shown in Table 2. Both new derivatives are effective against leukemia.P 388. Particularly interesting is the result obtained in the treatment of mice suffering from malignant tumors with compound XOO-0.63: at an optimal non-toxic dose of 15 mg/kg, more than 50% of the mice (6/10) are cured with compound XOO-0163 and at a dose of 10 mg/kg, 3/10 mice are cured and a significantly longer lifespan is achieved compared to the control group (205%). Therefore, the anti-tumor effect of compound XOO-0163 is definitely greater than that of compound DX, which at an optimal non-toxic dose of 10 mg/kg only treats a random malignant tumor in mice.

Compound XOO-0138 was also tested against Gross leukemia (treatment on day 1 after tumor inoculation). The data presented in Table 3 show that at the optimal non-toxic dose of 11.8 mg/kg, compound XOO-0138 exhibits antitumor activity that surpasses that of DNR and is similar to that of compound DX.

Table 1

Effect on HeLa cells ⁸ )

Compound Dose (mg/ml) % Dose (mg/ml) daunorubiein 12.5 7 7 6.25 53 3.1 110 compound XOO-0138 100.0 0 15 (4 ^, -deoxy-4-iododaunorubicin) 25.0 5 6.2 109 doxorubicin 25.0 0 8 ’2.5 16 6.2 70 compound XOO-0163 25.0 0 5.5 (4*-deoxy-4*-iodoxorubiein) 6.2 16 1.5 91 0.39 104

^a ) treatment for 24 hours *

Table 2

Effect against P 388 ascites leukemia ⁸ )

Compound Dose ^b ) (mg/kg) T/C ⁽ %) LTS ^d ) Toxic death·) daunorubiein 2.9 160, 165 0/10,0/10 0/10,0/10 4.4 160, 185 0/10,0/10 1/10,0/10 6.6 175, 135 0/10,0/10 7/10,7/10 compound XOO-0138 4.4 140 0/10 0/10 6.6 155 1/10 0/10 10.0 185.75 0/10,0/10 0/10,6/10 15.0 70 0/10 8/10 22.5 60 0/10 10/10 doxorubicin 4.4 210 0/10 0/10 6.6 260 0/10 0/10 10.0 265 1/10 0/10 compound XOO-OI63 6.6 240 0/10 0/10 10.0 305 3/10 0/10 15.0 >620 6/10 0/10 22.5 390 3/10 1/10

Explanations to Table 2:

^a ) Experiments were performed on BDP 1 or CDF 1 mice, which were inoculated intraperitoneally with 10® leukemia cells.

®) Treated intraperitoneally on day 1 after tumor inoculation.

^c ) Median survival time of treated mice/median survival time of control group x 100.

^(d ) Long-term survival (longer than 60 days).

^e ) Evaluated based on autopsy findings.

Table 3

Effect against Gross leukemia")

Compound Dose®) (mg/kg) T/C ^c ) * LTS ^d ) Toxic death*) daunorubicin 15.0 191 0/8 0/8 22.5 158 0/8 4/8 doxorubicin 7.7 150 0/10 0/10 10.0 175 0/10 0/10- 13.0 200 0/10 0/10 16.9 233 0/10 0/10 22.0 266 0/10 1/10 compound XOO-0138 11.8 208 0/10 0/10 ’5.4 208 0/10 2/10 4iododaunorubicin 20.0 175,117 0/8,0/10 4/8,6/10 30.0 100 0/8 8/8 40.0 92 0/8 8/8

^a ) C3H mice received intravenous injection of 2 x 10® leukemia cells. ®) Treated intravenously on day 1 after tumor inoculation.

^c ) See Table 2.

^d ) See Table 2.

*) See table 2.

Claims (1)

The invention A process for producing an antitumor active anthracycline glycoside compound of formula I wherein R represents a hydrogen atom or a hydroxy group, in the form of the hydrochloride, characterized in that 4'-deoxy-4'-iodo-N-trifluoroacetyldaunorubicin (II) dissolved in acetone is subjected to alkaline hydrolysis with 0.1 N aqueous sodium hydroxide solution under mild conditions for 3 hours at 0 ° C to give the free glycoside base of formula I, wherein R is hydrogen, which is isolated as a hydrochloride by treatment with a methanolic hydrochloric acid solution and optionally reacted dissolved in a mixture of dioxane and methanol, solution of bromine in chloroform to give the 14-bromo derivative, from which treatment with an aqueous solution of sodium formate at room temperature for 24 hours yields a glycoside compound of formula I wherein R is hydroxy, which is then isolated as the hydrochloride.