SU677293A1 - Actonocyl-bis-( -dimethylaminopropyl)-amine displaying antitumoral activity - Google Patents

Abstract

Actinocylbis- (G-dimethylamino-propyl) -amine of the formula: The invention relates to a new compound, an analogue of the natural antibiotic Actinomycin D, a large number of actinocyl bis-amino acids and their esters have been described at this time. Among these compounds, no compounds with practically valuable properties have been found to date. It is known that sarcolysin is active in the group of alkylating antitumor compounds. But along with antitumor activity in relation to certain transplantable tumors, sarcolysin is highly toxic. The purpose of the invention is to expand the range of biologically active compounds. According to the invention, actinocyl bis (y-dimethyl "nopropyl) -amine formula

Description

(CH3) gHCHH2CH2CHNHKN

WITH

which is obtained by the oxidation of potassium ferricinide in phosphate-alkaline buffer (pH 7.2) or PbO in alcohol.

a solution of 2-amino-3 hydroxy- -methylbenzoyl- (0 -dimethylaminopropyl) amine; the latter is obtained by catalytic hydrogenation of 2-nitro-3-benzyloxy-ethylbenzoyl (V-dimethylaminopropyl) amine or 2-nitro-3-hydroxy--methylbenzoyl- ( -dimethylaminopropyl) -amine. Example; 2-nitro; hydrochloride; 3-benzyloxy-4-methylbenzoyl (U-dimethylaminopropyl) amine.

To a solution of 5.6 g of 2-nitro-3-benzyloxy-t-methylbenzoic acid chloride in 100 ml of benzene, a solution of 1.77 g | ((dimethylaminrpropylamine in 20 ml) is added dropwise with stirring under stirring

benzene. Withstand reactionary sucks

Su while stirring at room temperature for 1 hour and then for 7 hours at 5060 ° C. A heavy precipitate is formed. After the end of the reaction,.

- the mass is evaporated to dryness. The dry residue is treated with 50 ml of benzene, heated to 70 ° C, cooled, the precipitate is filtered off, washed with a small amount of benzene, and dried.

Output. 4.2 g (56.3%). M.p. 1b6-1b8 ° C;

(isopropyl alcohol-benzene, 1:10). Chromatographically homogeneous on plates Silufol UV-.5 in the system 1-butanol-acetic acid-water (4: 1: 1). . Found,%: C 58.2; .H 6.5; C1 8.8. Calculated: C 58.6; And 6.4; C1 8,7 2-Nitro-3-benzyloxy-4-methylbenzoyl- (U-dimethylaminopropyl) -amine. . 3.5 g of 2-nitro-3-benzyloxy-H-methylbenzoyl- () -dimethylaminopropyl) -amine hydrochloride are dissolved in 20 ml of water, a saturated solution of sodium bicarbonate is added and extracted with ethyl acetate (3 x 20 ml). The ethyl acetate extracts were combined, washed with water, dried with Mai sulphate and evaporated in vacuo. Obtain 2.8 g of oily product, hroHHN% SNgN. K (CH3) 2

About Shg

matographically homogeneous on plates. Silufol UV-254 in the systems: 1-butanol-acetic acid-water (4: 1: 1),

chloroform-methanol-ethyl acetate (6: 1: 3). Analyze in the form of reinecate, which is obtained by mixing a solution of 2nitro-3-benzyloxy-4-methylbenzoyl (-dimethylaminopropyl) -amine in O, 1 n.

hydrochloric acid and an aqueous solution of ammonium reinecatte. The precipitated 2-nitro-3 benzyloxy-4-methyl-benzoyl- (3-dimethylaminopropyl) -amine reinecate is filtered off, washed with water and dried in a desiccator over. . -: - ... Found: C 41.6; H 4.6; Cr9.0, .N 17.9o. . ....; ..

С2оН25-% 04 C Il7CrN60} SS};

- Calculated,%: C 41.6; K-4.7;

Cr 9.3; N 18, D ...

2-Nitro-3 hydroxy-4-methylbenzoyl (y-dimethylaminopropyl) .- amine ..

. 0.5 g of 2-nitro-3 hydroxy-4-methylbenzoic acid is dissolved in 10 ml

absolute ether, add 20 ml of anhydrous benzene and to the resulting solution at room temperature and stirring add a solution of 0.52 g of dicyclohexylcarbodiimide in

5 ml of benzene The dicyclohexylurea is filtered off for 0.5 h, and a solution of 0.1 ° g of y-dimethylaminopropylamine in 5 ml of benzene is added dropwise to the filtrate while stirring. Stand under stirring for 0.5 h, the precipitate is filtered off, washed with a small amount of benzene and ether, dried, treated with 30 ml of water,

the aqueous solution is filtered.

evaporated to a volume of 1-2 ml and applied to a column containing., g cellulose LEAZ. Chromatograph in water, collecting a fraction corresponding to 35 of those on Silufol UV-254 plates in a methanol-ethyl acetate-triethylamine system (5: 5: 0 2-nitro-3-hydroxy-4-methylbenzoyl- (/ -pimethylaminopopip) -amine 5 The eluate is evaporated to dryness, Crystallized from a mixture of alcohol-benzene and from benzene to give 0.15 g.-Nitro-3-hydroxy-methylbenzoyl- SU-dimethylaminopropyl) -amine, mp, 182.5-1 hS, Found, С55.5; Nb, 9o Calculated: С 55,5; 6,8, Actinocyl-bis- (X-dimethylaminopropyl) -amine,. a) 2, p. g of 2-nitro-3-benzyloxy-methylbenzoyl- (X-dimethylaminopropyl) amine is hydrogenated in 100 ml of methanol at atmospheric pressure in the presence of palladium black to 2-amino-3-hydroxy-4-methylbenzoyl- (U-dimethylaminopropyl) -amine. Control by TLC on plates with Silufol yu-25 in the systems: 1-butanol-acetic acid-water) and chloroform-methanol-ethyl acetate. (6: 1: 3). At the end of the hydrogenation, the reaction mass is filtered, and 3.5 g of lead dioxide in 30 ml of methanol is added to the suspension with stirring and with stirring. The mixture is stirred at room temperature for 7 m, left for 12 hours, evaporated to dryness, extracted with chloroform, the chloroform extract is evaporated in a vacuum to dryness, and the residue is crystallized with benzene CZ. Output 1 g chromatographic; ; ; physically homogeneous Silufol UV-25; the system is chloroform-methanol-ethyl acetate (HR);}. actinocyl bis - (- dime. tylaminopropyl) -amine. M.p. 136-137 ° c. ; ;:. ::.;:;: - -;. //. ..) Filtered from catalysis. torus solution obtained with hydri. Rinse 0.55 g of 2-nitro-3-benzyloxy-methylbenzoyl- (3-dimethylamino-propyl. -amine), mixed with 100 ml of phosphate; alkaline buffer (pH 7.2) is added to a solution of 1.51 g of potassium ferricyanide in 15 ml of the same buffer. Stir for 3 hours at room temperature a saturated solution of sodium bicarbonate is added to the reaction mass and extracted with chloroform. The chloroform form extract is washed with water, dried with sodium sulfate, and evaporated to dryness. The residue is crystallized in benzene. g of substance with mp 135-137 ° C. Does not give depressions of the melting point in the sample sheni with a sample obtained by method a). The chromatographic mobility of samples obtained by methods a) and b) is the same. 3 c) Actiiocyl-bis - (- dimethylaminopropyl) -amine is prepared in a manner similar to METHOD b), starting from 2-NITRO-3-OXY-4-methylbenzoyl- (-dimethylaminopropyl) amine, which hydrogenates to 2-amino-3-hydroxy--methylbenzoyl- u-di-. methylamines) propyl) a-Mina is carried out at atmospheric pressure in ethanol in the presence of nickel P | 5ne. The product obtained by method c) is identical to the compound obtained by methods a) and b). . :.:. :: Found,%: C, 62.8; H 4.3; N 16.9 .. C26 "36% (4 -: -::. Calculated, C 62.9; H 4.3; N 16.9, - l.: .-.: ..... .. UV spectrum (sodium phosphate buffer, PH 6), them (Е) (2, Р, 10.), /, 45 (2, -10) ..:: ... ..; ,,.: .. .- Spectrophotometric -titration of actinocyl bis (y-dimethylaminopropyl) -amine with deoxyribonucleic acid (DNA) in sodium phosphate buffer (| mP, t) showed that the resulting compound complexed with DNA (Cb-U, B0.6) .. V. /;. V -:. The study of antitumor activity was carried out in vitro in the system of primary one-day stationary suspension cultures of tumor cells of mice (Table.) And in the system of primary 1X cultures of human tumor cells (Table 2), as well as in vitro in experiments on mice (Table 3). From the data presented in Tables 1 and 2, it follows that actinocyl-bis- (dimethylaminopropyl) -amine is It has an antitumor activity in these primary suspension cultures of mouse tumor cells and primary human tumor cells in vitro. The data given in Table III, indicate that actinocylbis (U-dimethylaminopropyl) amine exhibits an antitumor activity equal to the activity of actinomycin 1, when studying in. vitro on. solid tumor. (adenocarcinoma AK 755). It causes inhibition of the growth of this tumor. On 7P, whereas Akinomycin D inhibits the growth of this tumor by 63%. At the same time, unlike crinomycin D, actinocyl bis - (- diethylaminopropyl) amine is not effective in leukemia, and with a solid tumor it has a prolonged effect .; The maximum tolerated dose of acinocyl-bis- (y-dimethylaminopropyl) - 7 amine is 25 PO times the dose of actinomycin D, which indicates its lower toxicity. /. -.

Vine, µg / ml, suppressing synthesis amount; nucleic acids on. 677293; 8 Thus, actinocylbis (- "/ dimethylaminopropyl) -amine is a drug. less toxic and more selectively acting than actinomycin D, T 3 b l and c a 1

:: ::. :. - T a b and c a 2 Comparative data 6 cytostatic activity. , . primary tumors of man. . . . . Comparative data on

75.5

AK 755 L 1210 75.5

100.5 125.5

71, B 13.5, 0

no no no no v. ; . T a b l; and C a 3 antitumor activity of AK 755 0.03.5 Actinomycin D L 12.10. 0.12.2 La0.12.2

ten

677293

Continuation of Table 3 t 1 63,017.0 1.0 - no:; (after 9b h) 2bnet (after 96 h) 1 3net