9,10-DIHYDROACRIDINE DERIVATIVES POSSESSING ANTIMICROBIAL ACTIVITY

Abstract: The invention concerns new compouds of formula (1) wherein R=H or CH3 and of formula (2) wherein R=H or CH3, HX represents an acid, n is 1 or 2. The compounds possess antimicrobial activity.

Description: 9,10-dihydroacridine derivatives possessing antimicrobial activity The piesent invention concerns new derivatives of 9 10-dιhydroacπdιne 3 , 6-dιamιno- 10-methyl-9, 10-dιhydroacπdιne, 3 , 6-bι s(dιmethylamιno)- 10-methyl-9 10- dihydroacπdine, salts of 3,6-dιammo-10-methyl-9, 10-dιhydroacπdιne and salts of 3,6- / /s(dιmethylamιno)-10-methyl-9,10-dιhydroacπdιne These compounds possess antimicrobial activity Properties and reactivity of 10-methyl-9, 10-dιhydroacπdιne aie known fi om the monograph Heterocyclic Compounds, Vol 9 (Acπdines), ed R M Acheson Intersci Publ New York 1973, and from the Journal of Physical Chemistry 1991, 95 10240 3,6-Dιamιno-l O-methylacπdιnιum and 3 6-6/s(dιmethylamιno)-10-methyl- acndinium salts are known from the monograph Heterocyclic Compounds, Vol 9 (Acπdines), ed R M Acheson, Intersci Publ , New York 1973 The salts possess antibacterial activity According to the present invention there are provided the chemical compounds 3,6-dιamιno-10-methyl-9, 10-dιhydroacπdιne and 3,6-6/s(dιmethylamιno)- 10-methyl- 9,10-dιhydroacπdιne of the formula (1) wherein R = H oi R = CH In polai solvents these compounds undergo autooxidation to 3,6-dιamιno-l O-methylacιιdιnιum or 3 6- Z>/s(dιmethylamιno)-10-methylacπdιnιum salts, respectively The invention also pi ovides salts ot 3,6-dιammo-10-methyl-9 10-dιhydι oacπdιne and ι 6-/j/s(dιmethylamιno)-10-methyl-9, 10-dιhydroacπdιne of the toimula (2) wheiein R = H or R = CH„ respectively, HX represents organic or inorganic acid, wheieas n is 1 oi 2 The salts undergo autooxidation to 3,6-dιamιno-lO-methylacπdιnιιιm oi 3 6- /?/Λ'(dimethylamino)-10-methylacridinium salts, respectively, in water or alcohol solutions The invention provides a process for the manufacture of 3,6-diamino- 10-methyl- 9, 10-dihydroacridine and 3,6-£/s(dimethylamino)-10-methyl-9,10-dihydroacridine of the formula (1), which comprises reacting 3,6-diamino-10-methyl-acridinium or 3,6- t /5(dimethylamino)-10-methylacridinium salt, respectively, preferably chloride, bromide, methylsulphate or nitrate, with a reducing agent, preferably sodium borohydride or sodium ditionite, at the temperature of 0 to 20°C over a period of 1 to 5 hours while stirring intensively the reaction mixture According to the invention the reducing agent as a solid or as a solution in water is added to the solution of 3,6-diamino-10-methylacridinium or 3,6-/ /.v(dimethylamino)- 10-methylacridinium salt in water or methanol-water mixture containing 10 to 60 vol % of methanol. The concentration of the salt is in range of 0 05 to 0 25 mole/1 It is preferred that 1 to 3 moles of the reducing agent per 1 mole of the acridinium salt is added at the temperature 0 to 10°C The resulting precipitate is filtered off, dried in an argon atmosphere and under reduced pressure, and then extracted with chloroform, ethylene chloride or ethyl ether. The extracting solvent is evaporated under reduced pressure and the remaining solid product dried in an argon atmosphere and under reduced pressure using phosphorus pentoxide The invention also provides a process for the manufacture of salts of 3 ,6-diamino- 10-methyl-9, 10-dihydroacridine and 3 ,6-/>Λv(dimethylamino)- 10-methyl- 9, 10-dihydroacridine which consists in a reaction of 3,6-diamino- 10-metylacridinium salt or 3.6-A/s(dimethylamino)-10-methylacridinium salt, respectively, preferably chloride, bromide, methylsulphate or nitrate, with a reducing agent, preferably sodium borohydride or sodium ditionite, in water or methanol-water mixture containing 10 to 60 vol % of methanol, at the temperature 0 to 20°C Preferably 1 to 3 moles of the reducing agent, as a solid or as a water solution, is added at 0 to 10°C to the solution containing 0 05 to 0 25 mole/1 of the acridinium salt The product of the reduction reaction is isolated, purified by extraction with chloroform, ethylene chloride or ethyl ether, then dried in an argon atmosphere and under reduced pressure According to the invention the dry product of the reduction reaction is then allowed to react with water solution of an acid The reaction is carried out at the temperature 10 to 20°C by adding the solid reduction product to water solution of the acid which concentration is in range of 0 05 to 0 40 mole/1 It is preferred that the molar ratio of the acid to the reduction product is in the range of 1 to 4 When the reaction is completed the water is distilled off under reduced pressure, and the remaining solid product is vacuum dried using phosphorus pentoxide Alternatively, according to the invention the dry product of the reduction reaction is dissolved in an alcohol, preferably n-propanol, methanol, or ethanol To the resulting solution, which concentration is in range of 0 01 to 0 05 mole/1, a water solution of the acid is added It is preferred that the concentration of the acid is in range of 0 1 to 2 0 mole/1 and 1 to 4 moles of the acid per 1 mole of the reduction product is used The resulting precipitate is filtered off, washed with an appropriate alcohol and then with ethyl ether, and dried under reduced pressure using phosphorus pentoxide The invention is illustrated but not limited by the following Examples Example 1 Preparation of 3, 6-ώamιno-10-met yl-9, 10-dιhydroacrιώne A solution of 4 g of sodium borohydride in 80 ml of water was added over a period of 20 minutes to a solution of 10 g of 3.6-diamino-10-methylacridinium chloride in 200 ml of methanol-water mixture (1 1 v/v) The reacting mixture was stiπed at 5°C during the addition of sodium borohydride and for an additional half an houi thereafter, then the temperature was raised to 20 - 25°C and the stirring continued foi 1 hour The precipitated product was filtered off, washed twice with cold (5 - 10°C) water and dried using phosphorus pentoxide, first in an argon atmosphere and then under the pressure of 2 - 4 minHg The dry product was extracted with chloroform (4 x 200 ml) at room temperature, then the solvent was distilled off under reduced pressure at the temperature <20°C The remaining powder was dried using phosphorus pentoxide, first in an argon atmosphere and then under the pressure of 2 - 4 mmHg 3.6 g of 3,6-diamino- 10-methyl-9, 10-dihydroacridine of the formula ( 1 ) wherein R = H, were obtained as light-orange needles which melted and decomposed at 127°C The solubility of the product was good - in alcohols, acetonitrile, acetone, fairly good - in chloroform, ethers, benzene and very low - in water Spectroscopic properties of the compound obtained were as follows: 85 UV (methanol): λ max 305 nm (ε max 10 000) Η NMR (CDC1 3 ): δ ppm 3.26 (s, 3H, -CH 3 ), 3.56 (bs, 4H, NH 2 ), 3.69 (s, 2H, CH 2 ), 6.20 (s, 2H), 6.20 - 6.31 (dd, 2H), 6.84 - 6.94 (d, 2H). 3,6-Diamino-10-methyl-9, 10-dihydroacridine obtained as described above 90 undergoes autooxidation to 3,6-diamino-10-methylacridinium cation in polar solvents. 3,6-Diamino-10-methyl-9, 10-dihydroacridine obtained as described above possesses a strong antimicrobial activity which was tested and documented for the following bacteria: Staphylococcits aureus, Escherichia coli, Proteus vulgar is, Pseudomonas aeruginosa, Bacillus cereus and Bacillus subϊi/is, fungi: Candida 95 albicans and Trichophyton mentagrophytes, and viruses: Hepatitis B (HBV) and Herpes simplex typel (HSV-1). Example 2 Preparation of 3, 6-bis(dimethylamino)-10-melhyl-9,10-dihydroacridine 100 A solution of 3 g of sodium borohydride in 60 ml of water was added over a period of 30 minutes to a solution of 8 g of 3,6-/>/s(dimethylamino)-10-methylacridinium methylsulphate in 300 ml of methanol-water mixture (1 : 3 v/v). The reacting mixture was stirred at 5°C during the addition of sodium borohydride and for an additional half an hour thereafter, then the temperature was raised to 20 - 25°C and the stirring 105 continued for 1 hour. The precipitated product was filtered off, washed three times with cold (5 - 10°C) water and dried using phosphorus pentoxide, first in an argon atmosphere and then under the pressure of 2 - 4 mmHg. The dry product was extracted with chloroform (5 x 150 ml) at room temperature, then the solvent was distilled off under reduced pressure at the temperature <20°C. The remaining powder was dried 1 10 using phosphorus pentoxide, first in an argon atmosphere and then under the pressure of 2 - 4 mmHg. 3,2 g of 3,6-i/ ' .v(dimethylamino)-10-methyl-9, 10-dihydiOacridine of the formula (1) wherein R = CH 3 were obtained as orange crystals which melted and decomposed at 103°C. The solubility of the compound obtained was: good - in alcohols, acetonitrile, acetone, chloroform and fairly good - in ethers, benzene. Spectroscopic properties of the compound obtained were as follows: UV(methanol): λ max 308 nm (ε max 11000) Η NMR (CDC1 3 ) δ ppm 2.93(s, 12H, N(CH 3 ) 2 ), 3.39(s, 3H, N-CH 3 ), 3.73(s, 2H, CH 2 ), 6.26 - 6.36(dd, 4H), 6.97 - 7.00(d, 2H). 3,6-/^/ ' .s'(dimethylamino)-10-methyl-9, 10-dihydroacridine obtained as described above undergoes autooxidation to 3,6-J7Λ'(dimethyloamino)-10-methylacridinium cation when dissolved in methanol, acetone, acetonitrile, and also as a water suspension. 3,6- J /3/Λ'(dimethylamino)-10-methyl-9,10-dihydroacridine obtained as described above possesses a strong antimicrobial activity, which was tested and documented for the following bacteria: Staphylococcus aitreus, Escherichia coli, Proteus vulgaris, Pseudomonas aeruginosa, Bacillus cereus and Bacillus subtilis, and fungi: Candida albicans and Trichophyton mentagrophytes. Example 3. Preparation of the tartrate of 3, 6-diamino-10-methyl-9, 10-dihydroacridine A solution of 4 g of sodium borohydride in 80 ml of water was added over a period of 20 minutes to a solution of 10 g of 3,6-diamino-10-methylacridinium chloride in 200 ml of methanol-water mixture (1 : 1 v/v). The reacting mixture was stirred at 5°C during the addition of sodium borohydride and for an additional half an hour thereafter, then the temperature was raised to 20 - 25°C and the stirring continued for 1 hour. The precipitated product was filtered off, washed twice with cold water and dried using phosphorus pentoxide, first in an argon atmosphere and then under the pressure of 2 - 4 mmHg The dry product was extracted with chloroform (4 x 200 ml) at room temperature, then the solvent was distilled off under reduced pressure at the temperature <20°C. The remaining powder was dried using phosphorus pentoxide, first in an argon atmosphere and then under the pressure of 2 - 4 mmHg. Then 2.25 g of the reduction product was added, over a period of 10 minutes, to 50 ml of water solution containing 0.40 mole/1 of tartaric acid while stirring at room temperature. The reacting mixture was stirred for the additional 5 minutes and then watei was distilled off under vacuum at the temperature <30°C The remaining crystalline product was dried using phosphorus pentoxide at 20 - 30°C under the pressure of 0 1 mmHg 5 15 g of the tartrate of 3,6-diamino-10-methyl-9, 10-dihydroacι idιne of the formula (2) wherein R = H, HX = HOOC(CHOH) 2 COOH and n = 2 were obtained as light-orange crystals which melted and decomposed at 156°C The solubility of the compound was good - in water and fairly good - in methanol, ethanol, acetonitrile The spectroscopic properties of the compound obtained were as follows UV (water) λ ™ , 294 nm (ε max 10 800) 1H NMR (D 2 O) δ ppm 3 40 (s, 3H, CH.), 3 93 (s, 2H, CH 2 ), 4 58 (s, 4H, CHOH), 7 31-7 35 (s+d, 4H), 7 44-7 47 (d, 2H) Example 4 Preparation of the sulphate of 3,6-dιamιno-10-methyl-9, 10-dιhydroacrιdιne A solution of 4 g of sodium borohydride in 80 ml of watei was added ovei a period of 20 minutes to a solution of 10 g of 3,6-diamino-10-methylacridinium chloride in 200 ml of methanol-water mixture (1 1 v/v) Then the procedure of the Example 1 was followed 2 25 g of 3,6-diamino-10-methyl-9.10-dihydroacridine obtained was dissolv ed in 300 ml of methanol To this solution stirred at 10°C. 33 3 ml of water solution containing 0 3 mole/1 of sulphuric acid was added ovei a peπod of 10 minutes The reacting mixture was stirred at 10°C for additional 5 minutes, then the pi ecipitated product was filtered off, washed with methanol and vacuum diied using phosphorus pentoxide 2 70 g of the sulphate of 3,6-diaιnino-10-methyl-9, 10-dihydroacπdine of the formula (2) wherein R = H, HX = H 2 SO 4 and n = 1 weie obtained as light-orange crystalls which melted and decomposed at 148°C The solubility of the compound was fairly good - in water and very low - in methanol, ethanol, acetonitrile The spectroscopic properties of the compound obtained wei e as follows UV (water) λ ma 294 nm (ε ma , 1 1 000) Η NMR (D 2 O) δ ppm 3 43 (s, 3H, -CH.), 3 97 (s, 2H, CH 2 ), 7 02-7 06 (s+d, 4H), 7 36-7 39 (d, 2H) Example 5 Preparation of the hydrochlortde of 3, 6-dιamιno-10-methyl-9, 10-dιhydroacrιdme 3,6-diamino-10-methylacridinium chloride was reduced according to the 180 procedure described in Example 1 Then 2 25 g of the reduction product was dissolved in 400 ml of dry n-propanol. To this solution stirred at room temperature, 10 ml of water solution containing 2 0 mole/1 of hydrochloric acid was added over a period of 10 minutes The precipitated product was filtered off, washed with n-propanol and then with anhydrous ethyl ether and vacuum dried using phosphorus pentoxide 185 2 62 g of the hydrochloride of 3,6-diamino-10-methyl-9, 10-dihydroacridine of the formula (2) wherein R = H, HX = HCl and n = 2 were obtained as light-orange crystals which melted at 191-193°C The solubility of the compound was good - in water, methanol, ethanol and fairly good - in propanol The spectroscopic properties of the compound obtained were as follows 190 UV (water) λ ma 294 nm (ε max 11 300) H NMR (D 2 O) 5 ppm 3 41 (s, 3H, CH 3 ), 3 94 (s, 2H, CH 2 ), 7 02-7 06 (s+d, 4H), 7 35-7 39 (d, 2H) Example 6 195 Preparation of the hydrochloride of 3,6-lns(dιmethylamιno)-10-melhyl-9, 10-dιhydro- acridwe A solution of 2 5 g of sodium borohydride in 50 ml of water was added ovei a period of 30 minutes to a solution of 6 g of 3,6-/ /^(dimethylamino)-10-methyl- acridinium methylsulphate in 250 ml of methanol-water mixture (1 3 v/v) Then the 200 procedure of the Example 2 was followed Then 2 80 g of the product of the reduction reaction was added, over a period of 5 minutes, to 105 ml of a water solution containing 0 20 mole/1 of hydrochloric acid while stirring at the temperature of 10 - 15°C The reacting mixture was stirred for the additional 5 minutes at the same temperature and then water was distilled off under 205 vacuum at the temperature <20°C. The remaining crystalline product was dried using phosphorus pentoxide under the pressure of 0 1 mmHg 3 40 g of the hydrochloride of 3,6-bis(dimethylamino)-10-methyl-9, 10- dihydroacridine of the formula (2) wherein R = CH , HX = HCl and n = 2 were obtained as light-orange hygroscopic crystals which melted and decomposed at 174- 176°C The solubility of the compound was good - in water, methanol, ethanol, fairly good - in acetonitrile The spectroscopic properties of the compound obtained were as follows UV (water) λ max 295 nm (ε mj 10 700) Η NMR (D 2 O) δ ppm 3 50 (s, 12H, CH,), 3 61 (s, 3H, CH 3 ), 3 94 (s, 2H, CH 2 ), 7 31-7 35 (s+d, 4H), 7 44-7 47 (d, 2H) Example 7 Preparation of the citrate of 3,6-bιs(dιmethylammo)- 10-methyl-9, J 0-dιhydroacrιdme 3,6-B/.s(dimethylamino)-10-methylacridinium methylsulphate was reduced according to the procedure described in Example 2 Then 2 80 g of the reduction product was added, all at once, to 120 ml of methanol solution containing 0 20 mole/1 of citric acid while stirring at the temperature of 10 - 15°C The reacting mixture was stirred for the additional 5 minutes at the same temperature and then water was distilled off under vacuum at the temperature <20°C The remaining crystalline product was dried using phosphorus pentoxide under the pressure of 0 1 mmHg 6 57 g of the citrate of 3,6-bis(dimethylamino)-10-methyl-9, 10-dihydroacridine of the formula (2) wherein R = CH 3 , HX = H0 2 CCH 2 C(OH)(CO 2 H)CH 2 CO 2 H and n = 2 were obtained as orange, very hygroscopic crystals which melted and decomposed at 78- 80°C The compound was soluble in water, methanol, ethanol, acetonitrile, and insoluble in ether, chloroform, benzene The spectroscopic properties of the compound obtained were as follows UV (w ater) λ„, ax 295 nm (ε ma 10 300) Η NMR (D 2 O) δ ppm 2 71-2 92 (q, 8H, CH 2 ), 3,28 (s, 12H, CH 3 ), 3,43 (s, 3H, CH.), 3,93 (s, 2H, CH 2 ), 7, 10-7, 17 (s+d, 4H), 7,35-7,38 (d, 2H), The salts obtained as described above undergo autooxidation to the corresponding 3,6-diamino-10-methylacridinium salts or 3,6-bis(dimethylamino)-10- methylacridinium salts in water or alcohol solutions. 240 The salts obtained as described above possess a strong antimicrobial activity which was tested and documented for the following bacteria: Slaphylococcus aureus, Escherichia coli, Proteus vulgaris, Pseudomonas aeruginosa, Bacillus cereus and Bacillus sublilis, fungi: Candida albicans and Trichophyton mentagrophytes, and viruses: Hepatitis B (HBV) and Herpes simplex typel (HSV-1). 245

Claims: Claims New 9, 10-dihydroacridine derivatives of the formula (1 ) wherein R = H or CH. New 9.10-dihydroacridine derivative according to claim 1 which is 3,6-diamino- 10-methyl-9, 10-dihydroacridine New 9, 10-dihydroacridine derivative according to claim 1 which is 3,6-Λ/ (dιmethyl- amino)- 10-methyl-9, 10-dihydroacridine New 9, 10-dihydroacridine derivatives of the formula (2) wherein R is H or CH,, HX represents an acid, n is 1 or 2 New 9.10-dihydroacπdine derivatives according to claim 4 which are salts of 3, 6-dιamino-10-methyl-9, l 0-dihydroacridine New 9,10-dihydroacridine derivatives according to claim 4 which are salts of 3,6-/)7s(dimethylamino)-10-methyl-9, l 0-dihydroacridine AMENDED CLAIMS [received by the International Bureau on 19 June 1998 ( 19.06.98) ; original claims 1-3 cancel led ; remaini ng claims unchanged ( 1 page) ] 4. New 9, 10-dihydroacridine derivatives of the formula (2) wherein R is H or CH 3 , HX represents an acid, n is 1 or 2. 5. New 9,10-dihydroacridine derivatives according to claim 4 which are salts of 3 ,6-diamino- 10-methyl-9, 10-dihydroacridine. 6. New 9,10-dihydroacridine derivatives according to claim 4 which are salts of 3 ,6->/5(dimethylamino)- 10-methyl-9, 10-dihydroacridine. Statement under Article 19(1) As a consequence of the International Search Report the claims 1 to 3 have been cancelled. This cancellation may have the following impact on the description: Lines 2 and 3. Names: "3 ,6-diamino- 10-methyl-9, 10-dihydroacridine, 3 ,6-3/s(dimethylamino)- 10-methyl-9,l 0-dihydroacridine" should be removed. Line 9. The following text should be added: "A possibility of the formation of 3, 6-diamino-10-methyl-9,l 0-dihydroacridine in an electroreduction process is mentioned in Journal of Electroanalytical Chemistry, 1992, 338, 133-134." Lines 13 to 17. All the lines should be removed. Line 18. The word "also" should be removed. Lines 24 to 40. All the lines should be removed. Line 41. The word "also" should be removed. Example 1 and Example 2. These examples should be removed. Example 3 - should be renumbered to Example 1. In the line 142 the following text should be added: "3,6 g of 3,6-diamino-10- methyl-9,10-dihydroacridine of the formula (1) wherein R = H, were obtained as light-orange needles which melted and decomposed at 127°C." Example 4 - should be renumbered to Example 2. In the lines 162 and 163 the text: 'Then the procedure of the Example 1 was followed" should be replaced by the text: "Then the procedure described in the first part of the Example 1 was followed to give 3,6 g of 3,6-diamino-10-methyl- 9, 10-dihydroacridine." Example 5 - should be renumbered to Example 3. In the line 181 after the words "described in" the following text should be added: "the first part of the". Example 6 - should be renumbered to Example 4. In the line 200 the text: " Then the procedure of the Example 2 was followed" should be replaced by the text: "The reacting mixture was stirred at 5°C during the addition of sodium borohydride and for an additional half an hour thereafter, then the temperature was raised to 20 - 25°C and the stirring continued for 1 hour. The precipitated product was filtered off, washed three times with cold (5 - 10°C) water and dried using phosphorus pentoxide, first in an argon atmosphere and then under the pressure of 2 - 4 mmHg. The dry product was extracted with chloroform (5 x 150 ml) at room temperature, then the solvent was distilled off under reduced pressure at the temperature <20°C. The remaining powder was dried using phosphorus pentoxide, first in an argon atmosphere and then under the pressure of 2 - 4 mmHg. 3,2 g of 3,6-3/ ' ^dimethylamino)-10-methyl-9,10- dihydroacridine of the formula (1) wherein R = CH 3 were obtained as orange crystals which melted and decomposed at 103°C." Example 7 - should be renumbered to Example 5. In the line 220 the text: "in Example 2." should be replaced by the text: "in the first part of Example 6 to give 3, 6-/3/5(^dimethylamino)-10-methyl-9,l 0-dihydro- acridine."

Inventors: ADAMUS JAN, CIEBIADA IRENEUSZ, DENYS ANDRZEJ, GEBICKI JERZY, ADAMUS, JAN, CIEBIADA, IRENEUSZ, DENYS, ANDRZEJ, GEBICKI, JERZY
Applicants: ADAMUS JAN, CIEBIADA IRENEUSZ, DENYS ANDRZEJ, GEBICKI JERZY, POLITECHNIKA CODZKA, WOJSKOWA AKADEMIA MEDYCZNA, ADAMUS, JAN, CIEBIADA, IRENEUSZ, DENYS, ANDRZEJ, GEBICKI, JERZY
Application year: 1998
Main classification: C07D219
IPC codes: C07D219