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CHEMICAL PROBLEMS 2020 no. 2 (18) ISSN 2221-8688
UDC 54.057;54.061;615.074;615.076.7
SYNTHESIS AND BIOLOGICAL ACTIVITY OF NEW DITHIOCARBAMATE
DERIVATIVES
C.Y. Shukurov
Azerbaijan Medical University, Department of Pharmaceutical Chemistry 167, S. Vurgun str., Baku, AZ 1022, Azerbaijan; e-mail: china.86@mail.ru
Received 15.01.2020 Accepted 13.05.2020
Abstract: Nowadays there are various antifungal agents. These included many substances, including derivatives of dithiocarbamates. The article aims to synthesize new derivatives of dithiocarbamate and study their antifungal activity. To analyze and identify synthesized substances and determine their structural formulas, the following spectral methods were used: IR (Shimadzu 8400 FTlR), 1H NMR (Bruker 500 MHz UltraShield) in DMSO-d6 (internal standard - tetramethylsilane), Mass spectroscopy (Agilent 1100 Series LC /MSD Trap VL & SL). Elemental analysis (C, H vs N) was performed using a Perkin Elmer analyzer. The biological activity of synthesized derivatives of dithiocarbamates was explored and 100^g/ml dilution of compound J2 (2-((4-morpholinophenyl)amino)-2-oxoethyl 4-benzylpiperazine-1-carbodithioate) showed identical effect against Candida albicans, Candida crusei, Candida parapsilosis as fluconazole. Compound A4 (2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-2-oxoethyl-4-methylpiperazine-1-carbodithioate) showed the same activity against Candida parapsilosis and Candida glabrata as a standard drug. Keywords: dithiocarbamate derivatives, antifungal activity, antibacterial activity DOI: 10.32737/2221-8688-2020-2-174-180
Introduction
Today there are many diseases caused by microorganisms. Some of these diseases are caused by the spread of pathogenic fungal infections. There are approximately 1.5 million different species of fungi on Earth, but just 300 of them are known to make people sick. Fungal diseases are often caused by fungi that are common in the environment [1].
A series of N-mono- and N, N-disubstituted dithiocarbamates were investigated as inhibitors of three b-carbonic anhydrases (CAs, EC 4.2.1.1) from fungal pathogens Cryptococcus neoformans, Candida albicans and Candida glabrata, that is, Can2, CaNce103 and CgNce103, respectively. The new class of b-CA inhibitors may have potential for developing antifungal agents with a diverse mechanism of effect as compared to clinically used drugs for which drug resistance was reported [2].
Вrug resistance for establishing
antifungals, such as azole derivatives (fluconazole and ketoconazole), is a driving force in global mortality due to fungal infection. Identification of new molecular entities structurally unrelated to the above may represent a valid strategy to overcome resistance to currently available drugs. In an effort to develop highly potent antifungal agents, we report herein a series of 27 compounds of dithiocarbamate and rhodanine molecules containing pyridine moiety and their antifungal activity. Among synthesized compounds, several analogues showed potent antifungal activity [3].
A new series of Schiff bases of 4-(4-aminophenyl)-morpholine were synthesized and described by IR, 1H-NMR, 13C-NMR, mass spectral and elemental analyses. The compounds were screened for antibacterial (Staphylococcus aureus (ATCC 9144), Staphylococcus epidermidis (ATCC 155), Bacillus cereus (ATCC 11778), Micrococcus luteus (ATCC 4678),
CHEMICAL PROBLEMS 2020 no. 2 (18)
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Escherichia coli (ATCC 25922)), antifungal (Candida albicans (ATCC 2091) and Aspergillus niger (ATCC 9029)) activities [4-7].
Antibacterial activity was tested against Staphylococcus aureus (MTCCB 737), Pseudomonas aeruginosa (MTCCB 741), Streptomyces epidermidis (MTCCB1824) and Escherichia coli (MTCCB1652), as well as antifungal activity against Aspergillus fumigatus nigra Aspergillus.
A series of substituted piperazine derivatives were synthesized and tested for antimicrobial activity. Also, the antibacterial activity was tested against Staphylococcus aureus (MTCCB 737), Pseudomonas aeruginosa
(MTCCB 741), Streptomyces epidermidis (MTCCB1824) and Escherichia coli (MTCCB1652), and antifungal activity against Aspergillus fumigatus, Aspergillus flavus and Aspergillus niger. All synthesized compounds showed significant activity against bacterial strains but were found to be less active against tested fungi. In vitro toxicity tests revealed that some compounds showed lesser toxicity against human erythrocytes [5].
The purpose of our work is to take into the consideration of the main directions of research in the creation of new active, antifungal substances -drugs of the future - including derivatives of dithiocarbamates.
Materials and methods
The substances cited were used as starting materials for synthesis of A4, A5, J1 and J2 compounds: N-methyl piperazine aniline (Great Britain Maybridge), 4-methyl piperazine, 4-ethyl piperazine, carbon disulfide, chloroacetyl chloride (Sigma Aldrich), sodium hydroxide, ethanol, 4-morpholinoaniline (Great Britain Acros organics), 4-methyl 4-benzyl piperazine and 4-benzyl piperazine, tetrahydrofuran (THF) as solvent and triethylamine (TEA) as catalyst.
To identify and analyze synthesized compounds and their structural formulas, spectral research methods were used: IR (Shimadzu 8400 FTiR), 1H NMR (Bruker 500MHz UltraShield) in DMSO-d6 (internal standart - tetramethylsilane) Mass Spectroscopy (Agilent 1100 Series LC / MSD Trap VL & SL). Elemental analysis (C, H va N) was performed using Perkin Elmer analyzer.
Results and discussion
Synthesis of new derivatives of dithiocarbamates with morpholine moiety was performed in three steps: acetylation of 4-morpholineaniline with chloroacetyl chloride, synthesys of 4-methyl 4-piperazine-1-dithiocarbamate and 4-benzylpiperazine-1-dithiocarbamate sodium, final
synthesis is reaction of acetylated 4-morpholineaniline with the synthesized 4-methyl 4-piperazine-1-dithiocarbamate and 4-benzylpiperazine-1-dithiocarbamate sodium salts through obtaining basic substances (Scheme 1).
R=Ri=
- substance Jl; R=Ri=
substance J2.
Scheme 1. Final step of the synthesis of dithiocarbamate dervatives (J1 and J2 compounds).
Synthesis of new derivatives of dithiocarbamate salts. Final synthesis was the
dithiocarbamates with piperazine moiety was reaction of acetylated 4-piperazineaniline with the
performed in three steps: acetylation of N- synthesized 4-methylpiperazine-1-
methylpiperazineaniline with chloroacetyl dithiocarbamate and 4 ethylpiperazine-1-
chloride, synthesys of 4-methylpiperazine-1- dithiocarbamate salts by the obtaining of basic
dithiocarbamate and 4 ethylpiperazine-1- substances (Scheme 2).
R=Ri=CH3- substance A4 ; R=Ri=C2H5- substance A5 Scheme 2. Final step of the synthesis of dithiocarbamate derivatives (A4 and A5)
Substance J1:
2-((4-morpholinophenyl)amino)-2-oxoethyl-4-(4-methylbenzyl)piperazine-1-carbodithioate:
Chemical formula: C25H32N4O2S2. Yield: 79 %. Melting point 145.5°C. IR (KBr vmax cm-1): 3292 (N-H), 2852-2767 (Aliphatic C-H), 1666.5 (c=O), 1514-1473 (C=N va C=C), 813 (1,4-disubstituted benzene cycle). 1H NMR (500 MHz, DMSO-ûfe): S 2,3-4.63 (24H, m, Aliphatic protons), 6.89 (2H, d, Aromatic protons, J=9.04 Hz), 7.14 (2H, d, Aromatic protons, J=7.88 Hz), 7.20 (2H, d, Aromatic protons, J=7.92 Hz), 7.43 (2H, d, Aromatic protons, J=9.03 Hz), 10,04 (H, s, NH). MS(ES): M+1:422. Element analysis: Calculated (%); C, 56.97; H, 7.41; N, 16.61. Found (%); C, 56.78; H, 7.32; N, 16.54.
Substance J2:
2-((4-morpholinophenyl)amino)-2-oxoethyl-4-benzylpiperazine-1-carbodithioate: Chemical
formula: C24H30N4O2S2. Yield: 82 %. Melting point 148°C. IR (KBr vmax cm-1): 3292 (N-H), 2852-2767 (Aliphatic C-H), 1666.5 (C=O), 15141474 (C=N va C=C), 814 (1,4-disubstituted benzene cycle). 1H NMR (500 MHz, DMSO-^6): S 2,26-4.37 (24H, m, Aliphatic protons), 6.89 (2H, d, Aromatic protons, J=9.02 Hz), 7.15 (2H, d, Aromatic protons, J=7.88 Hz), 7.21 (2H, d, Aromatic protons, J=7.90 Hz), 7.43 (2H, d, Aromatic protons, J=9.07 Hz), 10,04 (H, s, NH). MS(ES): M+1:471. Element analysis: Calculated (%); C, 61.25; H, 6.42; N, 11.90. Found (%); C, 61.06; H, 6.35; N, 11.79.
Substance A4:
2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-2-oxoethyl-4-methylpiperazine-1-carbodithioate:
Chemical formula: C19H29N5OS2. Yield: 81 %. Melting point 196 °C. IR (KBr vmax cm-1): 3254 (N-H), 2918-2848 (Aliphatic C-H), 1657 (C=O),
1514-1415 (C=N va C=C), 820 (1,4-disubstituted benzene cycle). 1H NMR (500 MHz, DMSO-d6): ô 2,23-4.21 (24H, m, Aliphatic protons), 6.88 (2H, d, Aromatic protons, J=8.99 Hz), 7.41 (2H, d, Aromatic protons, J=8.99 Hz), 10,03 (H, s, NH). MS(ES): M+1:408. Element analysis: calculated (%); C, 55.99; H, 7.17; N, 17.18. Found (%); C, 54.83; H, 7.02; N, 17.04. Substance A5:
2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-2-oxoethyl-4-ethylpiperazine-1-carbodithioate:
Chemical formula: C20H31N5OS2. Yield: 84 %. Melting point 184°C. IR (KBr vmax cm-1): 3264 (N-H), 2974-2933 (Aliphatic C-H), 1658 (C=O), 1508-1429 (C=N va C=C), 825 (1,4-disubstituted benzene cycle). 1H NMR (500 MHz, DMSO-d6): ô 2,29-4.21 (24H, m, Aliphatic protons), 6.88 (2H, d, Aromatic protons, J=9.05 Hz), 7.42 (2H, d, Aromatic protons, J=9.02 Hz), 10,03 (H, s, NH). MS(ES): M+1:422. Element analysis: Calculated (%); C, 56.97; H, 7.41; N, 16.61. Found (%); C, 56.78; H, 7.32; N, 16.54.
Biological activity screening Anticandidal assay
It should be noted that synthesized products were tested for their in vitro growth inhibitory activity against human pathogenic,
such as Candida albicans, Candida crusei, Candida parapsilosis and Candida glabrata. Fluconazole was used as a positive control factor. Anticandidal activity test was performed according to CLSI reference M27-A3 broth microdilution method [8]. Compounds were dissolved in DMSO. Further dilutions of the compounds and standard drug in test medium were prepared in the required quantities of 1600, 800, 400, 200, 100, 50, 25, 12.5, 6.25, 3.125 and 1.5625 (ig/mL concentrations with Mueller-Hinton broth and Sabouroud dextrose broth. In order to ensure that the solvent percent had no effect on yeast growth, a control test was also performed in calculated broth supplemented with only DMSO in the same dilutions used in our experiments and found inactive in culture medium. MIC50 and MIC parameters of synthesized compounds were calculated (Table 1.). Also, inhibition of new derivatives against Candida species was illustrated (Diagrams 1 and 2). Compound J2 showed the same effect against Candida albicans, Candida crusei, Candida parapsilosis as fluconazole. Compound A4 revealed the same effect against Candida parapsilosis and Candida glabrata as standard drug.
Table 1. MIC50 and MIC parameters of synthesized compounds.
Compound Fungi species Minimum inhibitory concentration 50 (MIC50) Results of minimum inhibitory concentration
2-((4-morpholinophenyl)amino)-2-oxoethyl-4-benzylpiperazine-1 -carbodithioate (J2) Candida albicans 49.6 ^q/ml 100 ^q/ml
Candida crusei 36.8 ^q/ml 100 ^q/ml
Candida parapsilosis 37.57 ^q/ml 100 ^q/ml
Candida glabrata 78.09 ^q/ml 200 ^q/ml
2-((4-morpholinophenyl)amino)-2-oxoethyl-4-(4-methylbenzyl)piperazine-1-carbodithioate (J1) Candida albicans 78.78 ^q/ml 200 ^q/ml
Candida crusei 77.15 ^q/ml 200 ^q/ml
Candida parapsilosis 74.08 ^q/ml 200 ^q/ml
Candida glabrata 77.16 ^q/ml 200 ^q/ml
2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-2-oxoethyl 4-methylpiperazine- 1-carbodithioate (A4) Candida albicans 76.71 ^q/ml 200 ^q/ml
Candida crusei 75.58 ^q/ml 200 ^q/ml
Candida parapsilosis 34.85 ^q/ml 100 ^q/ml
Candida glabrata 37.48 ^q/ml 100 ^q/ml
2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-2-oxoethyl 4-ethylpiperazine-1-carbodithioate (A5) Candida albicans 71.24 ^q/ml 200 ^q/ml
Candida crusei 76.93 ^q/ml 200 ^q/ml
Candida parapsilosis 35.44 ^q/ml 100 ^q/ml
Candida glabrata 78.57 ^q/ml 200 ^q/ml
Diagram 1. Inhibition of Candida species by the compound 2-((4-morpholinophenyl) amino)-2-oxoethyl 4-benzylpiperazine-1-carbodithioate (J2)
120
1,5625 3,125 6,25 12,5 25 50 100 200 400 BOO 1600
DilutionSjlg/ml
Against C. albicans — Against C.crusei AgainstC.parapsilosis - Against C.glabrata
Diagram 2. Inhibition of Candida species by the compound 2-((4-(4-methylpiperazin-1-yl) phenyl) amino)-2-oxoethyl 4-methylpiperazine-1-carbodithioate (A4)
Conclusion
New derivatives of piperazine-1-dithiocarbamate with morpholineaniline (J1 and J2) and with N-methyl piperazineaniline (A4 and A5) were synthesized and their structures identified. The biological activity of synthesized
derivatives of dithiocarbamates were examined, MIC50 and MIC parameters of synthesized compounds were calculated and reveal antifungal activity of the compounds J2 and A4.
References
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YENiDiTiOKARBAMAT TÖROMdLdRIMN SiNTEZi Vd BiOLOJi FOALLIGI
QY. §ükürov
Azsrbaycan Tibb Universiteti, sczagiliq kimyast kafedrasi AZ 1022, Baki, S.Vurgun küg., 167, e-mail: china.86@mail.ru
Hal-hazirda bir gox müxtslif göbslsk sleyhins vasitslsr vardir. Bunlarin arasinda ditiokarbamatlar törsmslsri ds var. Bu i§in msqssdi yeni ditiokarbamat törsmslsrinin sintezi vs antifunqal fsalliginin öyrsnilmssidir. Sintez olunmu§ maddslsrin tsdqiqi vs strukturlarinin müsyysnls§dirmsk ügün a§agidaki spektral metodlardan istifads edilmi§dir: DMSO-d6-da IR (Shimadzu 8400 FTiR), 1H NMR (Bruker
500 MHz UltraShield) (daxili standart - tetrametilsilan), Mass spektroskopiya (Agilent 1100 Series LC) /MSD Trap VL & SL). Element analizi (C, H vs N) Perkin Elmer analizatoru vasitssi ils aparilmiçdir. Sintez olunmuç ditiokarbamat torsmslsrinin bioloji fsalligi tsdqiq edilmiç, J2 birlsçmssinin 100 mkq / ml konsentrasiyada durulaçmada Candida albicans, Candida crusei, Candida parapsilosis çtamlarina qarçi flukonazol ils muqayissds eyni fsalliq numayiç etdirmiçdir. A4 maddssi, standart numuns ils (flukonazol) muqayissds Candida parapsilozu vs Candida glabrata çtamlarina eyni fsalligi gostsrmiçdir.
Açar sozjidr: ditiokarbamatlar torsmslsri, antifunqalfsalligi
СИНТЕЗ И БИОЛОГИЧЕСКАЯ АКТИВНОСТЬ НОВЫХ ПРОИЗВОДНЫХ
ДИТИОКАРБАМА ТОВ
Ч.Я. Шукюров
Азербайджанский медицинский университет, Мфедра фармацевтической химии AZ 1022, Баку, ул. С.Вургуна, 167, e-mail: china.86@mail.ru
В настоящее время существует множество различных противогрибковых средств, среди которых есть и производные дитиокарбаматов. Целью данной работы является синтез новых производных дитиокарбаматов и изучение их противогрибковой активности. Для анализа и идентификации синтезированных веществ и определения их структурных формул использовались следующие спектральные методы: ИК (Shimadzu 8400 FTiR), 1H ЯМР (Bruker 500 MHz UltraShield) в flMCO-d6 (внутренний стандарт - тетраметилсилан), Масс-спектроскопия (Agilent 1100 Series LC / MSD Trap VL & SL). Элементный анализ (C, H и N) проводили с использованием анализатора Perkin-Elmer. Была исследована биологическая активность синтезированных производных дитиокарбаматов и соединение J2 (2 - ((4-морфолинофенил) амино) -2-оксоэтил 4-бензилпиперазин-1-карбодитиоат) при концентрации 100 мкг / мл показало такую же активность в отношении Candida albicans, Candida crusei, Candida parapsilosis, что и флуконазол. Соединение A4 (2 - ((4-(4-метилпиперазин-1-ил) фенил) амино) -2-оксоэтил 4-метилпиперазин-1-карбодитиоат) проявило активность в отношении Candida parapsilosis и Candida glabrata, что и стандартное лекарственное средство. Ключевые слова: противогрибковые средства, производные дитиокарбаматов
