Synthesis and SAR studies of pyrazole-3

ACCEPTED MANUSCRIPT

This is an early electronic version of an as-received manuscript that has been accepted for publication in the Journal of the Serbian Chemical Society but has not yet been subjected to the editing process and publishing procedure applied by the JSCS Editorial Office. Please cite this article as I. Bildirici, A. Cetin, N. Menges, Y. Alan, J. Serb. Chem. Soc. (2018), https://doi.org/10.2298/JSC170313029B This “raw” version of the manuscript is being provided to the authors and readers for their technical service. It must be stressed that the manuscript still has to be subjected to copyediting, typesetting, English grammar and syntax corrections, professional editing and authors’ review of the galley proof before it is published in its final form. Please note that during these publishing processes, many errors may emerge which could affect the final content of the manuscript and all legal disclaimers applied according to the policies of the Journal.

J. Serb. Chem. Soc. 83 (0) 1–13 (2018) JSCS–4809

UDC Original scientific paper

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Synthesis and SAR studies of pyrazole-3-carboxamides and -thioureides including chiral moiety: Novel candidates as antibacterial agents

ISHAK BILDIRICI1, ADNAN CETIN2*, NURETTIN MENGES1 and YUSUF ALAN3 1

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Van Yuzuncu Yil University, Faculty of Pharmacy, Department of Pharmaceutical Chemistry, Van, Turkey, 2Muş Alparslan University, Faculty of Education, Department of Science, 49250, Muş, Turkey and 3Muş Alparslan University, Faculty of Science and Arts, Department of Biology, 49250, Muş, Turkey (Received 13 March, revised 6 December 2017, accepted 14 February 2018)

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Abstract: A series of tetra substitute pyrazole-3-carboxamides (3a-c) and pyrazole-3-carboxthioureides (6a-c) were synthesized and their structures were characterized by IR, NMR, and Elemental analysis. The antibacterial potential against specific Gram-positive and Gram-negative strains and antifungal activities of all novel compounds were investigated. The structure-activity relationships (SAR) studies and some theoretical parameters (CLogP, CMR, PSA, ESP) of the compounds were carried out on these two pyrazole derivatives. Pyrazole-3-carboxilate (2) was used for the synthesis of carboxamide derivatives. The reactions of pyrazole-3-isothiocyanate (5) with appropriate chiral amino alcohols were utilized for synthesizing of thioureide derivatives. Both of these pyrazole derivatives including chiral moiety were exhibited pronounced antibacterial activities. According to the present in vitro study, some of our promising compounds might be new candidates as new generation antibacterial drug.

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Keywords: biological activity; chiral amino alcohols; pyrazole; heterocyclic compounds. INTRODUCTION

Chiral structures are important target molecules in chemistry since they have significant properties such as catalyst,1 biological,2 and pharmaceutical,3 agricultural4 and industrial implications.5 Chiral property has brought a third dimension to the all of this field of science. On the other hand, the carboxamide derivatives bearing a chiral stereocentre have also extensive workspace. For example, chiral carboxamides which have amide bond were used as a transporter for the drug in pharmaceutical.6 Furthermore, compounds having thiourea were * Corresponding author. E-mail: [email protected] https://doi.org/10.2298/JSC170313029B

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known to exhibit various biological properties such as antibacterial,7 antifungal,8 antitubercular,9 antithyroid,10 antihelmintic,11 rodenticidal,12 insecticidal,13 herbicidal14 and plant growth regulator15 activities. However, some thiourea derivatives including chiral moiety not only can serve as catalysts 16 for the synthesis of optically active compounds, but also can be employed as medicines.17-19 Studies on biological activities of chiral thioureas have relatively rare and reported studies showed that they have broad spectrum of biological activities such as anti-HIV,20 anticancer21 and antiviral22 agents. Some of the biologically active chiral thiourea and amide derivatives are showed in Figure 1. Since the beginning of this century, pyrazoles which are an important scaffold of heterocyclic compounds have increasingly taken attention in the literature because of their wide-spread of study field, particularly for their biological activities. These compounds have been used in the development of agricultural products and in drug research since they have diverse biological activities.23, 24 Some known activities such as pharmaceutical, agricultural and biological activities of these compounds containing pyrazole ring system in the structure can be listed as high antihyperglycemic,25 analgesic,26 inflammatory,27 antipyretic,28, 29 anti-bacterial,30 and antidepressant.31 H 3C

Cl

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H 3C O

O O S H 2N

F F F Celecoxib

Cl Rimonabant

O N N Carbimazole

CH 3

H3C

N H

CH 3

N H

N

Anti-allergic

O

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Cl

CH 3 S

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N NH N N

N N

H

O H OH Glycocholic acid

OH

NH

HO OH O

NH

NO2

H

HO

Cl Cl Chloramphenicol

Fig. 1. Some of the important examples of pyrazole, chiral carboxamides and thioureas.

They were provided protection against plant pests,32 as insecticides33 and against fungal organisms.34 They were also been detected as hypotensive35 and anticancer.36, 37 Samples including synthetic pyrazole such as anti-rheumatic Celecoxib has been used in the treatment of inflammation38 and pain.39 Rimonabant has been used for the treatment of obesity,40 Difenzoquat has been exerted lethal effects against plant pests41 and Tartrazine lemon yellow dye is

SYNTHESIS OF NOVEL PYRAZOLE-BASED ANTIBACTERIAL AGENTS

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widely used as colouring agent in food in the United Kingdom and United States42 (Fig. 1). Studies on the implications of stereochemistry of antibacterial agents have high significance in medicinal chemistry.43 Additionally, antimicrobial and antifungal activities of pyrazole-3-carboxamides and -thioureides including chiral stereocentre have not been reported in the literature until now. We recently disclosed the design, synthesis, and antimicrobial activity of a series of novel carboxamide and thioureide derivatives including pyrazole scaffold.44 Encouraged by these successful efforts, we aimed both to expand this study by synthesis new pyrazole derivatives containing chiral moiety and to evaluate the antimicrobial potential against various Gram-positive and Gram-negative strains and antifungal activities of these novel compounds. Moreover, we aimed to investigate the structure-activity relationships (SAR) studies and some theoretical parameters of these newly synthesized derivatives. EXPERIMENTAL

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Materials and equipment 1 H (400 MHz) and 13C (100 MHz) NMR spectra were recorded on a Bruker DRX-400 high performance FT-NMR spectrometer, NMR spectra was obtained in solutions of DMSOd6 and CDCl3. Analytical TLC of all reactions was performed on Merck prepared plates. Infrared spectra were recorded on a Shimadzu IR-470 spectrophotometer. The elemental analyses were obtained with Carlo-Erba Model 1108 apparatus. Optical rotations were taken on a Perkin Elmer 341 Model polarimeter. Refraction indices were measured by using an Atago Abbe refractometer. The mass spectrum was measured on Thermo Scientific TSQQuantum Access LC/MS spectrometers. Compound 1 (mp 202-204 oC) and 2 (mp 180 oC) was obtained according to our previous studies.44 Analytical and spectral data of the synthesized compounds are given in Supplementary material to this paper.

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General procedure for synthesis chiral pyrazole-3-carboxamides (3a-c) Compound 2 (0.404 g, 1 mmole) dissolved in methanol (5 ml) was put to reaction pot and then the solution of chiral amino alcohols (1 mmole) (2-amino-2-phenylethanol, 2-amino3-methyl-1-butanol, 2-amino-1-butanol respectively) in methanol (5 ml) was added drop-wise to solution at room temperature for 2 hours. After addition, solid product was precipitated, filtered and it was washed diethyl ether. The obtained product was purified by silica gel column chromatography (n-hexane/ethylacetate, 7:1). General procedure for synthesis chiral pyrazole-3-thioureides (6a-c) Compound 1 (0.397 g, 1 mmole) was refluxed with excessive SOCl2 at 80 oC for about 7 h. Excessive SOCl2 was evaporated. Remaining oily product was purified in dry ether/cyclohexane mixture. As result, 4-benzoyl-1-(2,5-dimethylphenyl)-5-phenyl-1Hpyrazole-3-carbonyl chloride (4) (m.p. 145 oC) was obtained according to the literature. 44 The resulting acyl-chloride compound (4) (1 mmole) dissolved in anhydrous acetone (15 ml) and a solution of (1 mmole) ammonium thiocyanate in acetone (5 ml) were added the reaction pot. The reaction mixture was refluxed in a round-bottom flask equipped with condenser for 4 hours. The solvent was evaporated and the residue was washed with ether. The solid product

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was filtered. Then the crude product was crystallized from n-hexane/ether. At last, 4-benzoyl1-(2,5-dimethylphenyl)-5-phenyl-1H-pyrazole-3-carbonyl-isothiocyanate (5) (m.p. 159-160.5 o C) was obtained. 44 Compound 5 (0.219 g 0.5 mmol) dissolved in anhydrous acetone (5 ml) was added dropwise to the appropriate chiral amino alcohol (1 mmol) (2-amino-2-phenylethanol, 2amino-3-methyl-1-butanol, 2-amino-1-butanol respectively) in acetone (5 ml). This mixture was room temperature for 4-6 hours. The reaction mixture was poured onto ice-cold water. The solid product was precipitated and filtered, dried and the product was purified by crystallization in ether/hexane.

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Microorganisms and antimicrobial assays Although antimicrobial activities of the starting compounds 1 and 2 were previously investigated,44 we examined their antimicrobial activities again in order to compare all compounds (including starting and newly synthesized compounds) in the same media together. All samples (1, 2, 3a, 3b, 3c, 5, 6a, 6b, 6c) were separately tested against Enterobacter aerogenes ATCC 27859, Bacillus subtilis, Staphylococcus aureus 6538, Bacillus megaterium DSM 32, Pseudomonas aeruginosa 9027, Klebsiella pneumoniae RSKK 574, Escherichia coli, Candida albicans ATCC 10231, Yarrovia lipolytica and Saccaromyces cerevisiae 6538 fungi. All bacterial and fungal strains examined in the present study were supplied by Muş Alparslan University (Turkey), Microbiology Laboratory. Penicillin (10 mg), Amikacin (30 mg), Erythromycin (15 mg), Rifampicin (5 mg) and Ampicillin (10 mg) antibiotics were used as reference drugs. The antimicrobial activities of the samples were determined by well diffusion method.45, 46 For this purpose, bacterial and fungal strains were cultured overnight at 37 ◦C in nutrient agar and 25 ◦C in Sabouraud dextrose agar medium, respectively. 100 µL of suspension of test microorganisms, containing 1 × 108 colonyforming units CFU/mL of bacteria cells and 1 × 104 CFU/mL spores of fungal strains were spread on Nutrient agar and Sabouraud dextrose agar medium, respectively. Subsequently, the medium was poured into a Petri dish on a horizontally levelled surface. After the medium was solidified, 9 mm diameter wells per dish were made in the agar medium. Then 0.005, 0.01 and 0.02 g/mL doses of 1, 2, 3a, 3b, 3c, 5, 6a, 6b and 6c suspensions dispersed in DMSO as 50 mg/mL were loaded into the wells separately. The Petri dishes were incubated at 37 ◦C for 24 h for bacteria and at 25 ◦C for 48 h for fungal strains. The average zone diameters were measured by repeating the experiment at least for three times. RESULTS AND DISCUSSION

Chemistry

In the present study, we prepared two series of new chiral derivatives 3a–c and 6a–c, which contained a 1,4,5-trisubstitute-pyrazole-3-carbonyl moiety attached to the chirality centre. Initially, 4-benzoyl-1-(2,5-dimethylphenyl)-5phenyl-1H-pyrazole-3-carboxylic acid 1 was synthesized according to our newly published manuscript and the corresponding methyl 4-benzoyl-1-(2,5dimethylphenyl)-5-phenyl-1H-pyrazole-3-carboxylate 2 was obtained by heating the pyrazole-3-carboxylic acid 1 and methyl alcohol with catalytic sulfuric acide.44 Then, these compounds were derived from amino alcohol such as (R)-2amino-2-phenylethanol, (R)-2-amino-3-methyl-1-butanol, (R)-2-amino-1butanol, respectively. The chiral pyrazole-3-carboxamides compounds were


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synthesized by means of the reaction between pyrazole-3-carboxylate 2 and chiral amino alcohols in the room temperature (Scheme 1). As a result of this reaction, different new chiral pyrazole-3-carboxamides 3a-c obtained in overall yield 72-78 % (Scheme 1).

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Scheme 1. Synthesis of pyrazole-3-carboxamides functionalized chiral amines

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All new synthesized compounds were confirmed by analytical and spectral data (see experimental). In the case of compounds 3a-c, the correct structures were established by IR and 1H-NMR spectroscopies in which characteristic -NH absorption bands were observed 3325-3210 cm-1 in the IR spectroscopy. Protons of hydroxyl signals and NH protons of amide groups signals were observed at  3.53-2.50 ppm and at  8.42-8.34 ppm in 1H-NMR spectroscopy, respectively. In order to incorporate potent active pharmacophore to a single molecule, such as pyrazole, thiourea, chiral amino alcohol, we prepared a series of chiral pyrazolo-thiourea derivatives (6a-c) by the reactions of 4-benzoyl-1-(2,5dimethylphenyl)-5-phenyl-1H-pyrazole-3-carbonyl-isothiocyanate 5 with the above mentioned chiral amino alcohols (Scheme 2). Therefore, firstly the corresponding 4-benzoyl-1-(2,5-dimethylphenyl)-5-phenyl-1H-pyrazole-3carbonyl chloride 4 was obtained by heating the pyrazole-3-carboxylic acid 1 with excess SOCl2 and then compound 5 was synthesized the pyrazole-3carbonyl chloride with ammonium thiocyanate in acetone about 5 hours.44 The chiral thioureide compounds 6a-c were prepared by basic chemical procedure. 6a-c were synthesized via reaction heating 4-6 hours pyrazole-3-carbonylisothiocyanate 5 and corresponding chiral amino alcohol derivatives and as outlined in scheme 2 (overall yield = 60-80 %). These types of heterocyclic compounds including chiral structures are not found in the literature so far. Structure elucidations of 6a-c compounds were based on 13C-NMR spectroscopy.

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Benzoyl carbonyl (C=O) signals were observed at  196.8-194.3 ppm, thioamide (C=S) signals were observed at  185.6-180.3 ppm and amide carbonyl (C=O) signals were observed at  167.7-165.9 ppm.

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Scheme 2. Synthesis of chiral substituted thiourea derivatives 6 via pyrazole-isothiocyanate 5.

Biology

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Antibacterial activity. Table I presents the antimicrobial activities of 1, 2, 3a, 3b, 3c, 5, 6a, 6b and 6c compounds against gram positive bacteria (Bacillus subtilis, Staphylococcus aureus 6538, Bacillus megaterium DSM 32) and gram negative bacteria (Enterobacter aerogenes ATCC 27859, , Pseudomonas aeruginosa 9027, Klebsiella pneumoniae RSKK 574 and Escherichia coli ATCC 25922) as expressed Minimal Inhibitory Concentration (MIC). Representative inhibition zone images were showed in Fig. 2. No antifungal activity of the compounds were detected against fungal strains (Candida albicans ATCC 10231, Saccharomyces cerevisiae, Yarrovia lipolytica) (data not shown). TABLE I. The zones of inhibition (mm) of the materials with antibiotics against bacterial and fungi strains (MICa in µg/mL) a MIC: Minimal inhibitory concentration values with SEM= 0.02, b ( - ): Totally inactive (no inhibition)

Cmpd. 1

2

Dose µg/mL 5 10 20 5 10 20

Gram-positive bacteria B. subS. B. megatilis aureus terium a b 12 13 14 13 14 12 12 10 12 10

Bacteria Gram-negative bacteria E. aero- E. P. aeruK. pneugenes coli ginosa monia 15 12 14 15 13 16 17 16 12 14 14 12 12 12 12 12 14 16 16 16

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TABLE I. Continued

3c

5

6a

6b

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3b

5 10 20 5 10 20 5 10 20 5 10 20 5 10 20 5 10 20 5 10 20

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3a

Bacteria Gram-positive bacteria Gram-negative bacteria B. subS. B. megaE. aero- E. P. aeruK. pneutilis aureus terium genes coli ginosa monia 10 10 14 14 15 12 16 12 15 19 20 16 19 20 21 19 11 12 11 10 12 10 10 13 12 15 14 15 12 14 16 22 18 18 16 20 18 12 12 12 15 11 11 16 15 12 11 16 17 17 18 18 19 15 11 13 12 12 12 14 13 15 16 17 16 16 17 18 16 14 13 13 13 12 16 15 16 12 15 18 16 18 18 19 19 18 20 18 12 12 12 12 15 14 15 15 17 13 14 18 18 18 20 18 17 19 13 12 14 13 14 13 13 13 14 13 14 15 15 15 15 15 14 16 Positive Controls 20 21 25 27 19 19 19

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Cmpd.

Dose µg/mL

15 30

14

10

-

10

13

-

16

30

21

18

16

16

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8

19

10

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-

10

-

-

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5

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Erythromycin Amikacin Penicillin Ampicillin Rifampicin

The in vitro antimicrobial and antifungal activities of the compounds and positive controls were tested in a dose manner (5, 10 and 20 µg/mL) against ten human pathogenic microorganisms (Table I). The current results indicated that the synthesized of these compounds showed a broad spectrum of the antibacterial activities producing 10-22 mm inhibition zones. Among the tested compounds 3b and 6a exhibited the highest antimicrobial activities. All tested compounds showed higher antimicrobial activity compared to Amikacin and Rifamicin

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except compound 2. Compound 3b and 6a had similar or higher antimicrobial activity compare to Penicillin. B

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Fig. 2. Inhibition zone images of the 6a against A: Staphylococcus aureus 6538, B: Enterobacter aerogenes ATCC 27859, C: Bacillus megaterium DSM 32.

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Moreover, obtained data indicated strong antibacterial activity of pyrazole-3carbonyl derivatives, such as; 3a-c and 6a-c exhibited pronounced activity against B. subtilis. 3a, 3b exhibited pronounced activity against S. aureus. 3b, 3c, 6a and 6b exhibited pronounced activity against B. megaterium. 3a-c, 6a and 6b exhibited pronounced activity against E. aerogenes. 3a exhibited strong activity against E. coli. 3a, 3b and 6a exhibited pronounced activity against P. aeruginosa. Compound 3b exhibited the highest antimicrobial activity against Grampositive bacteria, followed by 3a and 3b. With regards to Gram-negative bacteria, compound 3a showed highest antibacterial activity followed by 3b and 3c. Compound 6a-c showed similar antibacterial activities against Gram-positive and Gram-negative bacteria. The antimicrobial activity potential pattern of compound 6a > 6b > 6c for both of Gram-positive bacteria and Gram-negative bacteria was found. In general all compounds had pronounced antimicrobial activities against bacteria strains. However, S.aureus and B.megaterium seems to be more resistant to compounds and positive controls tested among bacteria strains. There is a positive correlation between antimicrobial activity and concentrations tested as shown in Table I. All tested compounds were pyrazole derivatives with certain modifications. Among the tested compound groups of 3 and 6 had the highest antimicrobial activities. The pronounced antimicrobial activities of compound 3 and 6 might be explained by the presence of chiral amino alcohol moiety. Relatively higher antimicrobial activities of 3b and 6a might be due to the presence of isopropyl and phenyl groups. The structure-activity relationships (SAR) studies Synthesized compounds were tried against many different types of bacteria and fungi. Three Gram-positive and four Gram-negative bacteria were applied


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and some important activities on bacteria were found. Unfortunately, for all compounds, no inhibition was observed against all fungi. The main skeleton of synthesized molecule was N-(2,5-dimethyl-phenyl)-4-benzoyl-5-phenylpyrazole. Changeable units were selected as ester (compound 2), carboxamide (compounds 3a-c), thiocyanate (compound 5) and carboxthioureide (compound 6a-c). All synthesized molecules were prepared at the concentration of 10 mM in DMSO. The molecules with chiral unit showed the best activity against all bacteria. Chiral molecules which have stereogenic centre are important structures because bioactive molecules usually observe for the activities of enantiomers. Chiral molecules should play a dominant role in their interactions with bioactive substances. Moreover, they should be applied only to molecules that contain the stereogenic centre in close proximity to the bioactive centre of the molecule as our synthesised compounds were. Synthetic routes to chiral molecules can be properly turned into from achiral structures. Having chiral structure molecule were synthesized by chiral transferred, whereby chiral starting material such as chiral amino alcohol. The ester group should be derivative with some more potent and suitable groups. For this reason, compound 2 was substituted by (R)-amino-alcohol derivatives to obtain compounds 3a-c. The most active derivative of them was found to be 3a which bears a phenyl group on it, against for all bacteria except for S. Aureus and B. Megaterium. It was seen that bulky group on the (R)-aminoalcohol affected the activity against bacteria. To clear this idea, some physicochemical parameters were calculated.47 So, hydrophobic (CLogP: Calculated partition coefficient), steric (CMR: Calculated molecular refractivity) and electronic (PSA: Polar surface area) parameters for all synthesized compounds were calculated. Theoretical lipophilicity of 3a-c decreases from 3a to 3c and was calculated as 6.67, 6.14 and 5.77, respectively. Calculation of CMR and CLogP for 3a-c revealed that CMR and CLogP affect the antibacterial activity and increasing of them has good impact on the biological activity. The worst activity was detected for 3c which bears an ethyl group in which CMR and CLogP are 138.43 and 5.77, respectively (Table II). Polar surface areas for 3a-c were calculated and it was seen that there is no changing for compound 3a-c. Thiocyanate group attached to the molecule 5 shown lower activities than 3a-c. On the other hand, activities of 5 on bacteria were moderate. Compound 6a-c which have thiourea and (R)-amino-alcohol chains were tested against same bacteria and it was calculated that reactivity against bacteria decreased and the most potent molecule of compounds 6a-c was 6a which bears phenyl ring. Theoretical parameters of compound 6a-c were also shown that biological activity could be affected by increasing of CMR and CLogP (Table II). Moreover, although CMR and ClogP of 6c was very close to 3a which has the best potent molecule, biological activity of it is the lower than 3a. Decreasing of

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reactivity on bacteria have shown that (R)-amino-alcohol unit should be adjacent to the carbonyl group and there should be no any other chain between carbonyl and (R)-amino-alcohol groups. After SAR study, it was worthy to say that (R)amino-alcohol unit increase the biological activity against some Gram (-) and Gram (+) bacteria. TABLE II. Some theoretical parameters for synthesized molecules CMR (cm3/mol) 116.47 120.33 153.7 142.90 138.43 129.83 170 159 154

CLogP 5.03 6.31 6.67 6.14 5.77 6.90 6.83 6.38 6.02

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PSA (Ao2) 69.97 49.74 82 82 82 62 94 94 94

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Compound 1 2 3a 3b 3c 5 6a 6b 6c

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Electron surface potential (ESP) of 3a was investigated to understand of its H-donor and -acceptor units and its total electronic surface. According to ESP map, oxygen atoms of two carbonyl groups and of alcohol group are H-acceptor and proton of alcohol group is an H-donor group (Fig. 3).

Fig. 3. Electron surface potential (ESP) of 3a.

Proton of amide group is lesser acidic than the others and due to this situation, this proton might not be a good candidate for proton donation. The rest of molecule has shown expected electronic potential and it is not worth to discuss those surfaces. CONCLUSION

Within this study, a series of tetra substituted pyrazole-3-carboxamides (3ac) and pyrazole-3-carboxthioureides (6a-c) binding chiral amino alcohol were


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synthesized and their antibacterial and antifungal properties were investigated. The structure-activity relationships (SAR) studies and some theoretical parameters (CLogP, CMR, PSA, ESP) of these pyrazole-carboxamide and thioureide derivatives were investigated. Compound series of 3 and 6 exhibited superior antimicrobial activities among compounds tested. The pronounced antimicrobial activities of compound series of 3 and 6 can be explained by the presence of chiral amino alcohol moiety. Relatively higher antimicrobial activities of 3b and 6a might be due to the presence of isopropyl and phenyl groups. The present results indicated that the synthesized compounds were active in a broad spectrum against important human pathogenic microorganisms. Therefore these compounds might be new candidates of efficient antibacterial agents. SUPPLEMENTARY MATERIAL

Analytical and spectral data of the synthesized compounds are available electronically at the pages of journal website: http://www.shd.org.rs/JSCS/, or from the corresponding author on request.

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Acknowledgements. The authors thank to the Management Unit of Scientific Research Projects of Mus Alparslan University (MSÜBAP) for financial support under Project MSÜ14EMF-G05. Authors thank to Dr. Abdullah Dalar for his valuable comments and corrections in antimicrobial section. ИЗВОД

СИНТЕЗА И SAR ИСПИТИВАЊЕ ПИРАЗОЛ-3-КАРБОКСАМИДА И ТИОУРЕИДА И ХИРАЛНИХ СТРУКТУРА: НОВИ КАНДИДАТИ КАО АНТИБАКТЕРИЈКСИ АГЕНСИ ISHAK BILDIRICI1, ADNAN CETIN2, NURETTIN MENGES1 и YUSUF ALAN3

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Van Yuzuncu Yil University, Faculty of Pharmacy, Department of Pharmaceutical Chemistry, Van, Turkey, 2 Muş Alparslan University, Faculty of Education, Department of Science, 49250, Muş, Turkey и 3Muş Alparslan University, Faculty of Science and Arts, Department of Biology, 49250, Muş, Turkey

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Синтетисана је серија пиразол—карбоксамида (3a-c) и пиразол—карбокстиоуреида (6a-c) и добијеним дериватима структура је одређена ИЦ, НМР и елементалном анализом. Испитана је антибактеријска активност према специфичним грампозитивним и грам-негативним сојевима бактерија и антифунгална активност свих нових синтетисаних једињења. Урађена је анализа утицаја структуре на активност (structure-activity relationships, SAR) и неких теоријских параметара (CLogP, CMR, PSA, ESP). За синтезу карбоксамидних деривата коришћен је пиразол-3-карбоксилат (2). Реакцијом пиразол-3-изотиоцијаната (5) и одговарајућих хиралних алкохола добијени су деривати тиоурее. Обе групе добијених једињења показују запажену антибактеријску активност. Према приказаном in vitro испитивању, неки од деривата могу бити кандидати за даља испитивања антибактеријске активности. (Примљено 13. марта, ревидирано 6. децембра 2017, прихваћено 14. фебруара 2018)

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6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17. 18. 19. 20. 21.

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Synthesis and SAR studies of pyrazole-3-carboxamides and -thioureides including chiral moiety: Novel candidates as antibacterial agents

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ISHAK BILDIRICI1, ADNAN CETIN2*, NURETTIN MENGES1 and YUSUF ALAN3

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Van Yuzuncu Yil University, Faculty of Pharmacy, Department of Pharmaceutical Chemistry, Van, Turkey, 2Muş Alparslan University, Faculty of Education, Department of Science, 49250, Muş, Turkey and 3Muş Alparslan University, Faculty of Science and Arts, Department of Biology, 49250, Muş, Turkey J. Serb. Chem. Soc. 83 (0) (2018) 000–000

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(R)-4-benzoyl-1-(2,5-dimethylphenyl)-N-(2-hydroxy-1-phenylethyl)-5phenyl-1H-pyrazole-3-carboxamide (3a). Yield: 88 %. [α]D23 +142.0 (c, 0,001, EtOH), Rf = 0.38, m.p.: 210-212 oC; Anal. Calcd. for C33H29N3O3 (Mw=515.22) C, 76.87; H, 5.67; N, 8.15 %; Found: C, 76.71; H, 5.59; N, 8.30 %; IR (ν, cm-1): 3275, 3154, 3056, 2865, 1692, 1664, 1603, 1548, 1476, 1434, 1274; 1H-NMR (400 MHz, DMSO-d6,  / ppm): 8.42 (d, 1H, J= 9.4 Hz, NH), 7.91-7.82 (m, 3H, Ar-H), 7.61-7.52 (m, 5H, Ar-H), 7.24-7.15 (m, 6H, Ar-H), 6.63-6.56 (m, 4H, ArH), 4.55 (m, 1H, CH), 3.53-3.50 (m, 3H, OH and CH2), 2.17 (s, 3H, CH3), 1.76 (s, 3H, CH3); 13C-NMR (100 MHz, DMSO-d6,  / ppm): 192.4, 166.2, 145.3, 143.6, 141.3, 139.5, 137.6, 136.2, 134.5, 133.6, 132.3, 121.2, 129.7, 128.1, 123.6, 121.4, 116.5, 111.4, 63.2, 54.7, 21.6, 17.4; (+)ESI-HRMS m/z: calculated for [C33H29N3O3 + H+]: 516.2287; observed 516.2271. (R)-4-benzoyl-1-(2,5-dimethylphenyl)-N-(1-hydroxy-3-methylbutan-2-yl)-5phenyl-1H-pyrazole-3-carboxamide (3b). Yield: 85 %. [α]D23 -204.0 (c, 0,001, EtOH), Rf = 0.38; m.p.: 191-193 oC; Anal. Calcd. For C30H31N3O3 (Mw=481.24): C, 74.82; H, 6.49; N, 8.73 %. Found: C, 74.69; H, 6.61; N, 8.92 %; IR (ν, cm-1): 3312, 3168, 3040, 2818, 1696, 1668, 1576, 1482, 1401, 1370, 1315. 1H-NMR (400 MHz, DMSO-d6,  / ppm): 8.35 (d, 1H, J= 9.2 Hz, NH), 7.90-7.88 (m, 2H, Ar-H), 7.78-7.77 (m, 2H, Ar-H), 7.26-7.16 (m, 5H, Ar-H), 6.76-6.52 (m, 4H, ArH), 3.71-3.58 (m, 3H, CH and CH2), 2.50 (m, 2H, CH and OH), 2.17 (s, 3H, CH3), 1.61 (s, 3H, CH3), 1.42 (d, J= 7.2 Hz, 6H); 13C-NMR (100 MHz, DMSOd6,  / ppm: 194.2, 162.5, 144.1, 142.7, 141.5, 140.1, 138.6, 136.4, 135.3, 133.1, 132.0, 130.5, 129.1, 128.4, 127.6 122.3, 121.1, 109.2, 63.3, 55.3, 30.1, 21.0,

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* Corresponding author. E-mail: [email protected]

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18.4, 12.2; (+)ESI-HRMS m/z: calculated for [C30H31N3O3 + H+]: 482.2444; observed 482.2441. (R)-4-benzoyl-1-(2,5-dimethylphenyl)-N-(1-hydroxy-butan-2-yl)-5-phenyl1H-pyrazole-3-carboxamide (3c). Yield: 85 %. [α]D23 +186.3 (c, 0,001, EtOH), Rf = 0.39, m.p.: 199-201 oC; Anal. Calcd. For C29H29N3O3 (Mw=467.22): C, 74.50; H, 6.25; N, 8.99 %. Found: C, 74.71; H, 6.42; N, 8.73 %; IR (ν, cm-1): 3362, 3156, 3066, 1576, 1489, 1385, 1310; 1H-NMR (400 MHz, CDCl3,  / ppm): 8.34 (d, 1H, J= 9.2 Hz, NH), 7.88-7.74 (m, 2H, Ar-H), 7.52-7.45 (m, 5H, Ar-H), 7.257.14 (m, 5H, Ar-H), 6.77 (m, 1H, Ar-H), 3.82 (m, 2H, CH2-OH), 3.51 (m, 1H, CH-NH), 2.68 (s, 1H, OH), 2.13 (s, 3H, CH3), 1.87 (s, 3H, CH3), 1.62 (m, 2H, CH2), 1.35 (t, 3H, J= 7.1 Hz, CH3); 13C-NMR (100 MHz, CDCl3,  / ppm): 198.3, 164.7, 146.3, 143.2, 141.5, 134.1, 133.9, 132.9, 131.6, 130.8, 130.0, 129.5, 129.1, 128.1, 126.3,125.4, 121.1, 108.2, 58.6, 55.3, 30.1, 22.1, 17.3, 14.6; (+)ESI-HRMS m/z: calculated for [C29H29N3O3 + H+]: 468.2287; observed 468.2287. (R)-1-(4-Benzoyl-1-(2,5-dimethylphenyl)-5-phenyl-1H-pyrazole-3-carbonyl)3-(2-hydroxy-1-phenylethyl)-thiourea (6a). Compound 6a was prepared according to general procedure. Yield: 61 %. [α]D23 +192.3 (c, 0,005, EtOH), Rf = 0.37, m.p.: 231-233 oC; Anal. Calcd. For C34H30N4O3S (Mw=574.20): C, 71.06; H, 5.26; N, 9.75 %. Found: C, 71.23; H, 5.34; N, 9.79 %; IR (ν, cm-1): 3355, 3212, 3054, 2925, 1712, 1686, 1674, 1572, 1494, 1442, 1323. 1H-NMR (400 MHz, DMSO-d6,  / ppm): 8.29 (m, 1H, NH), 8.21-7.04 (m, 16H, Ar-H), 6.57 (m, 2H, Ar-H), 3.95 (m, 2H, CH2-OH), 3.13 (m, 1H, CH-NH), 2.30 (m, 1H), 2.17 (s, 3H, CH3), 1.18 (s, 3H, CH3); 13C-NMR (100 MHz, DMSO-d6,  / ppm): 198.3, 188.6, 166.7, 143.1, 141.2, 136.7, 135.4, 132.6, 132.2, 131.5, 131.1, 130.4, 130.2, 130.0, 129.9, 129.8, 129.6, 129.3, 129.2, 129.1, 128.7, 128.5, 128.1, 127.2, 122.5, 109.1, 64.2, 50.3, 21.4, 16.8; (+)ESI-HRMS m/z: calculated for [C34H30N4O3S + H+]: 575.2117; observed 575.2111 (R)-1-(4-Benzoyl-1-(2,5-dimethylphenyl)-5-phenyl-1H-pyrazole-3-carbonyl)3-(1-hydroxy-3-methylbutan-2-yl)-thiourea (6b). Compound 6b was prepared according to general procedure. Yield: 56 %. [α]D23 +125.4 (c, 0,005, EtOH), Rf = 0.37, m.p.: 207-209 oC; Anal. Calcd. For C31H32N4O3S (Mw=540.22): C, 68.87; H, 5.97; N, 10.36 %. Found: C, 68.95; H, 5.89; N, 10.42 %; IR (ν, cm-1): 3365, 3151, 3062, 2957, 1704, 1686, 1671, 1568, 1498, 1444, 1321. 1H-NMR (400 MHz, DMSO-d6,  / ppm): 8.12 (m, 1H, NH), 8.05-7.20 (m, 12H, Ar-H), 6.64 (m, 1H, Ar-H), 3.93 (m, 2H, CH2-OH), 2.88 (m, 1H, CH-NH), 2.41 (bs, 1H, OH), 2.21 (s, 3H, CH3), 1.95 (m, 1H, CH), 1.79 (s, 3H, CH3), 1.64 (m, 3H, CH3), 1.18 (s, 3H, CH3); 13C-NMR (100 MHz, DMSO-d6,  / ppm): 196.8, 180.3, 165.9, 144.7, 142.3, 141.8, 137.5, 135.2, 134.4, 133.2, 132.1,131.7, 129.8, 129.1, 128.6, 127.5, 126.3, 125.4, 124.2, 121.7, 108.2, 64.1, 59.7, 32.1, 21.4, 18.4, 17.2;

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(+)ESI-HRMS m/z: calculated for [C31H32N4O3S + H+]: 541.2273; observed 541.2269. (R)-1-(4-Benzoyl-1-(2,5-dimethylphenyl)-5-phenyl-1H-pyrazole-3-carbonyl)3-(1-hydroxy propan-2-yl)-thiourea (6c). Compound 6c was prepared according to general procedure. Yield: 50 %. [α]D23 +153.2 (c, 0,005, EtOH), Rf = 0.37, m.p.: 215-217 oC; Anal. Calcd. For C30H32N4O3S (Mw=512.19): C, 67.95; H, 5.51; N, 10.93 %. Found: C, 68.56; H, 5.91; N, 10.51 %; IR (ν, cm-1): 3435, 3165, 3058, 2895, 1711, 1685, 1667, 1572, 1498, 1442, 1323. 1H-NMR (400 MHz, DMSO-d6,  / ppm): 8.67 (bs, 1H, NH), 8.23-6.97 (m, 13H, NH and Ar-H), 6.83 (m, 1H, Ar-H), 3.74 (m, 2H, CH2-OH), 2.81 (m, 1H, CH-NH), 2.25 (m, 4H, OH and CH3), 2.13 (s, 3H, CH3), 1.66-1.63 (m, 2H, CH2), 1.13 (m, 3H, CH3); 13 C-NMR (100 MHz, DMSO-d6,  / ppm): 194.5, 181.0, 167.1, 144.4, 141.6, 139.5, 137.2, 136.7, 135.3, 131.5, 130.3, 129.9, 129.4, 129.1, 128.8, 128.6, 128.4, 128.0, 127.5, 124.3, 108.8, 62.3, 55.7, 27.3, 22.6, 17.1, 10.6; (+)ESIHRMS m/z: calculated for [C30H30N4O3S + H+]: 527.2117; observed 527.2110.

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