CS227686B2 - Manufacture of anti-tumor active anthracycline glycoside compounds - Google Patents

Abstract

The present invention relates to a method of making antitumor effective anthracycline glycoside a compound of formula (I)

Description

The present invention relates to a process for the production of an anti-tumor anthracycline glycoside compound of formula I

where.

R represents a hydrogen atom or a hydroxy group, characterized in that 4-deoxy-4-iodo-N-trifluoroacetyldaunorubicin of the formula 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 treated with a methanolic solution hydrochloric acid is isolated as the hydrochloride and optionally reacted dissolved in a mixture of dioxane and methanol with a solution of bromine in chloroform to give the 14-bromo derivative, from which treatment with an aqueous solution of sodium formate at room temperature after 84 hours yields the glycoside compound wherein R is hydroxy which is then isolated as the hydrochloride.

Both the novel glycosides, i.e., 4-deoxy-4-iodounorubicin and 4-deoxy-4-iodo-oxirubicin, obtainable by the method of the invention have excellent female anti-cancer cells.

The starting material for their preparation is - as mentioned above - 4'-deoxy-4-iodo-N-trifluoroacetyldaunorubicin. This compound is described in published European Patent Application Publication No. 81 10 7618.1 of September 24, 1981,

The novel anthracycline glycosides of the present invention exhibit anti-cancer activity. In particular, 4'-deoxy-4-iodo-oxorubicin (a compound of formula I wherein R is hydroxy) has a distinct effect in mouse cancer experiments.

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

The invention will now be described in more detail by means of production examples and biological data.

He did

4’-deoxy-4-ioddaunorubicin (1} R = Η) (XOO-0138)

A solution of 2.0 g of 4-iodo- N -trifluoroacetyldaunorubicin in 10 ml of acetone is 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 aglycones.

The aqueous solution was 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, m.p. 57 DEG C. (dec.). spectrum: 637 (M ^& lt ^{; +} & gt ^; ).

^TLC plate from ^F 254 silica gel (Merck ^firm ) using a 80: 20: 7: 3 mixture of chloroform, methanol, water and acetic acid had an Rf = 0.72.

Example 2

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

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

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 was evaporated in vacuo and the residue was dissolved in a mixture of 200 ml of chloroform and methanol, both solvents being in a volume ratio of 13: 6. The organic phase was 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 .alpha.-deoxy-.alpha.-iodo-oxorubicin hydrochloride having a melting point of 230 DEG C. (decomposition) is obtained; mass spectrum 653 (M ⁺⁾ and by thin layer chromatography (silica gel plate F254 ^{(MEROKA CIWS) using} a mixture of chloroform, methanol, water and acetic acid in a volume ratio of 80: 20: 7: 3, the R ^. = 0.5 .

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

Compounds were tested in comparison to daunorubicin (DNR) and doxorubicin (DX) in vitro against HeLa cells. The data in Table I show that XOO-0138 is less cytotoxic than DNR, while XOO-0.63 is more cytotoxic than DX. The anti-cancer effect 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. Of particular interest is the result obtained by treating a mouse suffering from cancer with compound XOO-0.63: at an optimal non-toxic dose of 15 mg / kg with compound XOO -0163 cures more than 50% of the mice (6/10) and at a dose of 10 mg / kg, 3/10 mice are cured and a significantly longer life expectancy 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 the optimal non-toxic dose of 10 mg / kg treats only a randomized cancer of the mice.

Compound XOO-0138 was also tested against Gross Leukemia (treatment on Day 1 after cancer inoculation). The data presented in Table 3 show that at the optimal non-toxic dose of 11.8 mg / kg of XOO-0138, it exhibits an antitumor effect that exceeds the effect of DNR and is similar to that of 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 Dec (4 ^, -deoxy-4-iodounorubicin) 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'-ioddoxorubiein) 6.2 16 1.5 91 0.39 104

^a ) 24-hour treatment *

Table 2

Effect against P 388 ascitiotic leukemia ⁸ )

Compound Dose ^b ) (mg / kg) T / C ^(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

Explanatory notes to Table 2:

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

®) Treatment intrapritoneally on day 1 after cancer inoculation.

^c ) Mean survival of treated mice / mean survival of control group x 100.

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

^e ) Evaluated on the basis of an autopsy finding.

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 4joddaunorubicin 20.0 175,117 0 / 8.0 / 10 4 / 8.6 / 10 30.0 100 ALIGN! 0/8 8/8 40.0 92 0/8 8/8

^a ) C3H mice were injected intravenously with 2 x 10 6 leukemia cells. ®) Treated intravenously on day 1 after cancer 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.