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Section 5. Chemistry
https://doi.org/10.29013/AJT-21-3.4-43-49
Mamedov Elnur Shamkhal oglu, senior research assistant, Institute of Catalysis and Inorganic Chemistry named after acad. M. Nagiyev of ANAS, Baku, Azerbaijan
E-mail: chem@kqki.science.az Veliyeva Dilara Safar kizi, senior research assistant, Institute of Catalysis and Inorganic Chemistry named after acad. M. Nagiyev of ANAS, Baku, Azerbaijan
Kulibekova Tamilla Nasraddin kizi, senior research assistant, Institute of Catalysis and Inorganic Chemistry named after acad. M. Nagiyev of ANAS, Baku, Azerbaijan
Rushinaz Irada ramiz kizi, research associate, Institute of Catalysis and Inorganic Chemistry named after acad. M. Nagiyev of ANAS, Azerbaijan Safaraliyeva Zubeyda Sabir kizi, research associate, Institute of Catalysis and Inorganic Chemistry named after acad. M. Nagiyev of ANAS, Baku, Azerbaijan
Huseynova Sara Enver kizi, research associate, Institute of Catalysis and Inorganic Chemistry named after acad. M. Nagiyev of ANAS, Baku, Azerbaijan
E-mail: chem@kqki.science.az
SYNTHESIS OF 6-METHYLURACIL DERIVATIVES AND THEIR PHARMACOLOGICAL ACTIVITY
Abstract. Selenium-containing derivatives of 6-methyluracil were synthesized, their reactivity was identified. Biological activity of uracils was studied, a simplified one-stage synthesis method was found and optimum conditions for obtaining these compounds were determined. It was established that synthesized compounds exhibit high antimicrobial activity.
Keywords: synthesis, selenium, 6-methyluracil, alkyl derivatives, diallyl, diallyl derivatives, biological activity.
One of the developing sections in synthetic organ- the numerous class of which uracils have a special ic chemistry is the chemistry of nucleosides, among place, that is due to the wide range of their properties.
At present, derivatives of the most studied representative of this class of 5-fluorouracil with various substituent have been synthesized, their reactivity and addition order of ligands were found out and their biological activity was studied [1-5].
Study of biological activity of uracils and their derivatives led to further development of potential medicinal products on their basis. Data from the authors [1-4,5], on the basis of which biological activity of uracils was determined by adding different functional groups into the position of "6" uracil ring, also is of great interest.
It has been proven by many developments that one of the active uracil derivatives is 6-methylura-
R=H,
The reaction for producing alkyl methyluracils was performed in one step by reacting 6-methylu-racil with alkyl halide and potassium carbonate in dimethylformamide solution. Triethylbenzeneam-monium chloride (TEBACH) and 18-crown-6-ether (18-C-6) were used as catalysts to synthesize all compounds. In the first case, all with other things being equal, yield of target compounds was less than in the second. Therefore, the experimental part describes the procedures for the synthesis of these compounds only with the participation of 18-crown-6-ether. Theoretically, alkylation of 6-methyluracil may proceed in two directions: in N1 and N3 positions. In our studies, alkylation proceeds according to the
cil and its derivatives which by accelerating the cell regeneration and having wide range of anabolic and anticatabolic activity and anti-inflammatory properties, form part of different medicals and are used as intermediate compounds in the synthesis of new medicinal products [5-13].
As a continuation of the works on the synthesis and development of substances with supposed biological activity, we obtained new alkyl derivatives of 6-methyluracil (I) (MeUr) [14]: 2- (6-methylura-cyl-1) -ethyl bromide (II); 1,3-diallyl-6-methylura-cil (III); 1-(2,3-epoxypropyl)-6-methyluracil (IV); 1-(2-oxy-3-chloropropyl) -6-methyluracil (V) by reaction of 6-methyluracil with alkyl halides.
(iii);
position N1 (i.e., the substituent is in position 1), by the data of NMR'H of synthesized compounds, proton signal of NH3 observed in the region of 10.2, that is consistent with literature data [15].
In view of the fact that uracils and their derivatives are of interest in synthesis of biologically active compounds on their basis, and also, considering highly interesting studies on the synthesis of selenor-ganic compounds with the same important properties, it seemed interesting to perform works on their combination and production of new compounds, also with potential bioactive properties.
For this purpose, some selenium-containing derivatives of 6-methyl-uracil were obtained. Thus,
R'=-CHCHBr(H);
R= R'=(CH2CHCH2)2
bis- (6-methyluracylyl-l) selenoxomethane (VI) tetrachloride and sodium selenide in the presence was synthesized by reacting the latter with carbon of copper monochloride as a catalyst.
Bis-[2-(6-methyluracylyl-l) ethyl] selenide (VII) enide (VIII) were obtained by reaction ofalkyl methy-and bis [3-(6-methyl- uracylyl-l) -2-oxypropyl] sel- luracils II and V with sodium selenide:
Bis-[(6-methyluracil-l)-2-bromopronyl]sel- alkylmethyluracil (IX) with selenium tetrabromide enedibromide (X) was synthesized by reaction of obtained by us earlier [10]:
MeUiCHiC H=CH2 + SeBn-KMeUiCI f2CI 1(Bi)C Fb]2SeBr2
Reaction mechanism [16] proceeds through: - the formation of transition complex (iXa) with its further conversion to [(6-methyluracil- l)-2-bro-mopropyl] selenium tribromide, which by covalency ofbond Se-Br it is ionized in solvent with formation of dibromo[(60methyluracilyl- l)]-2-bromopropyl selenonium bromide (iXb). The latter, being the source of the bromonium ion and the cation ionic intermediate [(6-methyluracilyl- l)-2-bromopropyl] selenium dibromide, joins the second molecule (IX)
to form an adduct (X). The reaction is carried out at a molar ratio (IX) to SeBr4 2:1.
Synthesized compounds (II-X) are light crystals, the structure of which has been proven by NMR and IR spectroscopy.
Experimental part
2-(6-methyluracilyl-1) ethylbromide (II).
3.8 g(20 mmol) of dibromoethane in 20 ml of dimethyl formamide (DMFA), 0.15 g (0.57 mmol) 18-C-6 is added into three-necked flask and 2.5 g
(20 mmol) of 6- methyluracil dissolved in 30 ml of DMFA is was poured dropwise at 300C. The temperature is raised up to 40 0C and the reaction mixture is stirred for 12 hours. After completion, the filtrate is dispersed. Precipitated crystals are washed with iso-propyl alcohol, sulfuric ether and dried. Ivory crystals are separated: Yield 1.48 g(37%). Tmelt _ 255-260 0C.
IR-spectrum v, cm1:1470 (CH2, CH3), 1690 (C = =O), 3195 (NH), 2865 (CH). NMR'H-spectrum ct, ppm: 10.2 m (NH), 5.45 d (1H, C(5) H), 3.65 t (2H, CH2Br).
1,3-diaIlyI-6-methyIuraciI (III). 2.6 g (20 mmol) of 6-methyluracil 1.38 g (10 mmol) of K2CO3, 0.15 g (0.57 mmol) of 18-C-6, 4.8g (40 mmol) of bromal-lylene, 30 ml of DMFA is poured into reaction flask and stirred for 6 hours at 500C. As the reaction proceeded, precipitation occurred and is decanted. It is
NMR'H ¡spectrum
dried, washed with hot heptane. White powder is separated: T > 350 0C (does not melt). Yield - 3.15 g (77.3%). IRspectrum v, cm-1: 1635-1650 (C=C al-lyl). 1695 (C=O) NMR'H spectrum ct, ppm: 7.74 d (1H, C(5) H), 5.60 m (2H, CH=C-), 5.22 m (4H, C-CH2), 44.40 d (2H, N1-CH2), 4.70 d (2H, NH3-CH2).
1-(2,3-epoxypropyl) -6-methyluracil (IV). 3.9 g(30 mmol) of MeUr is added into the flask, dissolved, 30 ml of dimethylformamide, 1.38 (10 mmol) of K2CO3, 0.15 g (0.57 mmol) of 18-C-6 and 2.8 g (30 mmol) of epichlorohydrine is added dropwise at 50 0C. After 6 hour stirring, the solution is distilled, crystalline yellowish oily precipitate remained at the bottom of the flask, is washed with hot isopropyl alcohol, and dried: yield - 0.7 g (41%). Tmelt.-270-272 0 C. IR-spectrum, v, cm-1:1720 (C=O), m100. 1250,
V o
ppm: 10.1 m (III, NH), 5.12 m (2H,
NC^),
OH
3.58 m (2H, C-CH-C), 2.15 m (2H, C - CH2)
V
1-(2-oxy-3-chloropropyl) -6-methylura-cil (V). A solution of 1.2 g of NaOH in 10 ml of water is added dropwise into a mixture of 3.78 g (30 mmol) of 6MeUr in 30 ml of DMFA at 500C, and is stirred for 2-3 hours. Without removing sodium salt of 6-methyluracil, 0.15 g (0.57 mmol) of 18-C-6 is added to the reaction flask, and 3.4 g (30 mmol) of dichloropropanol is poured dropwise. Stirring is continued for 6 hours at 70 0C. As the reaction proceeded, a white curd precipitate is deposited on the bottom of the flask. It is filtered in glass filter, washed with heptane and dried. Yield - 2.8 g (43%). Tmelt 283-285 0C. IR-spectrum, v, cm-1: 1730 (C=O), 1100, 3400 (OH), 3100 (N-H). NMR'H spectrum ct, ppm: 4.92 c (1H, Oh), 4.08 d (1H, C-CH-C), 3.62 d (2H, NCH2), 2.84 d (2H, CH2Cl).
Bis-(6-methyluracil)-1-selenoxomethane (VI). Mixture of2.56 g(20 mmol) of 6-methyl-uracil, 15.4 g(10 mmol), 0.1 g(1 mmol) cuprous chloride in
mixture of 20 ml of dimethylformamide and 20 ml of ethyl alcohol is heated at 60-650C for 6 hours. Upon completion of the reaction a precipitation occurs at the bottom ofa flask. The precipitate is filtered, washed with hot ethanol and dried. Yield - 1.7 g (51%). T 240-245 0C. IR-spectrum, v, cm1: 790, 1430, 1485 (C=Se), 1720 (C=O), 3145 (NH). NMR'H spectrum ct, ppm: 10.08 m (1H, NH), 7.5 (III, C(5), H).
Bis- [ (6-methyluracilyl-1) ethyl] selenide (VII). 4.7 g (20 mmol) of compound (II), 1.2 g (10 mmol) of Na2Se, 0.4 g (1 mmol) of 18-K-6 and 50 ml of absolute alcohol is poured into three-necked flask with nitrogen supply tube and mixed at 60 0C for 6 hours, filtered, the filtrate is vaporized. The precipitate is washed with chloroform and hot alcohol: yield 1.3 g(34%): Tmelt 298-300 0C. IR-spectrum, v, cm1: 730(C-Se), m480 (CH2CH2), 3190 (NH). NMR'H spectrum ct, ppm: 99.8 m (1H, NH3). 4.83 t (4H, CHSe), 4.36 t (4H, NCH2).
Bis-[3-(6-methyluracilyl-l)-2-oxypropyl] selenide (VIII). It is synthesized using the method of producing compound VII from 4.3 g (20 mmol) compound (V), 1.25 g (10 mmol) of Na2Se, 0.4 g (1 mmol) of 18-K-6 and 50 mol of absolute alcohol. The precipitate is washed with hot alcohol and chloroform: yield is 1.9 g (42%). Tmelt 181182 0C. IR-spectrum, v, cm-1: 735 (C-Se), 1720 (C=O), 1100, 3480 (OH). NMR'H spectrum a, ppm: 4.86 c (2H, CH). 3.96 m (2H, C-CH-C), 3.60 d (4H, CH2Se).
Bis-[(6-methyluracilyl-1) -2-bromopropyl] selendibromide (IX). 4 g (10 mmol) of SeBr4 in 10 ml of chloroform is dropped into the solution of 3.4 g (20 mmol) (IX) in 30 ml of chloroform for an hour under nitrogen. After mixing the mass for 3 hours at 30 0C, the precipitate is filtered, washed with hot chloroform: yield is 3.2 g (44%). Tmelt- 117-118 0C. IR-spectrum, v, cm-1: 740 (C-SeX 1680 (C=C), 1730 (C=O) NMR'H spectrum a, ppm: 4.38 t (2H, CHBr). 3.85 d (4H, CH2Se), 10.28 (2H, NH), 3.23 t (4H, NCH2).
Antimicrobial properties of (II-VI) Considering that molecules of synthesized compounds contain fragments that are part of many medicinal preparations, it seemed obvious to study their bioactive properties.
Tablel. - Antimicrobial actisvity
The table shows antimicrobial influence data of compounds (II-VI) on testing cultures: Staphylococcus aureus (Si-aureus), Escherichia coli (E. coli), Pseudomonas aeruginosa (Ps. aeroq.), and Candida albicans.
Antimicrobial activity testing of synthesized al-kyl-6-methyluracil, including its selenium derivative: (6-methyluracilyl-l)-ethylbromide (II), 1,3-dial-kyl-6-methyluracil (III), 1 - (2,3-epoxypropyl) -6-meth-yluracil (IV), 1-(2-oxy-3-chloropropyl)-6-methylu-racil (V), bis-(6-methyluracilyl-l) selenoxomethane (VI) was performed using the method of serial dilution. Tested substances were taken in the form of1% of alcohol solution. Their activities were studied in the following dilutions: 1:200, 1:400, 1:800, 1:1600.
Meat infusion agar was used as a nutrient medium with pH 7,2-7,4, but for Candida - Sabouraud's medium.
In all tests microbial load was 1 billion of microbial bodies per 1 ml, from which one drop was added to each tube. Sowing was performed every 10, 20, 40, 60 minutes of exposure. For comparative study of the activity of studied medicinal products known preparations were taken as a control (ethyl alcohol, furacin, nitro- fungine). It should be noted that all compounds in dilution of 1.1600 were inactive to the testing-cultures.
of 6-methyluracil derivatives
Expo- Dilution deg ree
№ Compound sure St.aureus E.coli Ps.acroq. Candida
time of sowing, min. 1:200 1:400 1:800 1:600 1:200 1:400 1:800 1:600 1:200 1:400 1:800 1:600 1:200 1:400 1:800 1:600
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19
10 + + + + — — + + — + + + — + + +
II 6MeUrCH2CH2Br 20 40 + + + + + + + + — — + + + + — — + + + + — + + + + + +
60 + + + + — — + + — — + + — + + +
10 - - + + + + + + — — + + — — + +
III 6MeUr(CH2CHCH2)2 20 40 — — — — + + + + + + + + — — + + + — — + + + +
60 - - — — + + + + — — — + — — + +
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19
6McUrCHiCTCHj 10 - — + + + + + + — — + + — — + +
IV 20 40 — — — + + + + + + + + + + — — + + + — — + + + +
60 - — — + + + + + — — — + — — + +
10 — — + + + + + + — — + + — — + +
V 6MeUrCH2CH(OH)CH2Cl 20 40 — — + + + + + + + + + + + + — — + + + + — — + + + +
60 — — + + + + + + — — + + — — + +
10 — — + + + + + + + + + + — — — +
VI [6MeUr] 2—C=Se 20 40 — — + + + — + + + + + + + + + + + + + + — — — + +
60 — — — + — + + + + + + + — — — +
In relation to S. aureus, tested compounds III, IV, V exhibit a similar bactericidal effect at 1:200, 1:400 dilutions after 10 min. of exposure. Compound III has the strongest antimicrobial effect at 1:800 dilution after 20 min.
Only compounds II and III have an effect on E. coli. at 1:200 and 1:400 dilutions after 10 minutes of exposure. The rest compounds did not exhibit bactericidal effect.
As to P. aeruginosa, all compounds exhibit similar effect at 1:200, 1:400 dilutions after 10 minutes
of exposure. Compounds III and IV has an effect at a greater dilution of 1:800, but only after 40 min. but compound. V has no bactericidal effect at all.
As to Candida, compound II was inactive - it exhibited only antimicrobial effect at 1:200 dilution after 10 minutes. The rest compounds (III-V) also have a destructive effect at 1:400 dilution. None of these compounds (II-V) has any effect on this testing culture at 1:800 dilution.
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