Microwave-Assisted Synthesis of New 4-Amino Acid Substituted
1,2,4-Triazole Derivative Derived from 1,2,3-Oxadiazole Nucleus and Their Anti-Bacterial and Anti-Oxidant Potential
Jaya Rautela1*, Ajay Singh Bisht2, Vikash Jakhmola1, A. N. M. Ansori3, Divya Negi4
1Uttaranchal Institute of Pharmaceutical Science, Uttaranchal University, Dehradun, India.
2Himalayan Institute of Pharmacy and Research, Dehradun, India.
3Professor, Nidom Foundation, Surabaya, Indonesia.
4Amrapali Institute of Pharmacy and Science, Haldwani, India.
*Corresponding Author E-mail: jayarautela01@gmail.com
ABSTRACT:
Heterocyclic compounds especially, five or six-member heterocyclic compounds, having one to three hetero-atoms in their nucleus take part in metabolism of all living cells. And this information gives the lead for synthesizing heterocyclic derivatives. This research work has focused on development of new method for synthesis of some new amino acid substituted 1,2,4-trizole derivative from 1, 3, 4-Oxadizole nucleus. The nucleus of the 1, 3, 4-Oxadizole was first produced by reaction of Isoniazide and 3-chlorobenzeldehyde which is then converted into 1,2,4-trizole derivative by the loss of water molecule. After that incorporation of different amino acid (luecine and glycin) leads new amino acid substituted 1,2,4-trizole derivative. The newly synthesized derivative Compound A1 and A2 were characterized by optical rotation, melting point, TLC, UV spectroscopy and IR. The antibacterial was screened using Disc diffusion method with ciprofloxacin as reference drug and antioxidant activity by in-vitro DPPH method using ascorbic acid as standard drug. Synthesized compound A1 showed maximum MIC against S. aureus as compared to Compound A2. For antioxidant activity, compound A1 showed maximum percentage inhibition as compared to Compound A2.
KEYWORDS: 1, 2, 4-Triazole, Anti-bacterial property, Antioxidant, IR, TLC.
INTRODUCTION:
Drug development is a highly controlled, diverse, creative, and inventive process1. And it is observed that with the help of the microwave-assisted reactions, revolution in organic compound synthesis can promoted2. Green Chemistry includes microwave assisted processes since they decrease reaction time, maximize percent yield, and are environmentally safe. Microwave radiations are types of electromagnetic energy, which lies at the lower end of the electromagnetic energy3. The basic principle behind Microwave-assisted reaction is lies on absorption of microwave energy by the reaction mixture, because of “microwave dielectric heating”.4
Some Heterocyclic compounds possess central role in pharmacology (antihypertensive, antiviral, antidepressants, and anticancer activity), agricultural (herbicides, and pesticides), and industrial (colorants) fields which increases value of heterocyclic compound in chemistry5.
As their name suggests, these are the compound having cyclic ring in their structure that contain at least one hetero (oxygen, nitrogen, and sulfur) atom in the ring6. Oxadiazole comes under aromatic heterocyclic molecules which contains N (nitrogen) with O (oxygen) in their five-member cyclic ring. The molecular formula of Oxadiazole is C2H2N2O7. Oxadiazole is available in four different isomers. Among all 1,3,4-oxadiazole possesses therapeutic importance, like Anti-tuberculosis activity, Anti-allergic activity, Anti-viral activity, Muscle relaxants activity etc8-9. The chemistry of Oxadiazole has been thoroughly investigated and is continuously being investigated. The -N=C-O- linkage has biological activity by reacting with the nucleophilic centers of malignant cells10. The ring skeleton has anticancer, antimicrobial, antiallergic anti-inflamatory, anti-fungal11-16. To appreciating the significance of 1,3,4-oxadiazole derivatives with amino acids In-Silico Estimation and Structural Docking Experiments are studied17.
Due to its broad biological action, there are already a number of inventions for the synthesis of substituted oxadiazole derivatives such as synthesis of a new family of 1,3,4-oxadiazoles bearing the benzimidazole moiety, new substituted-1,3,4-oxadiazol fluoro cinnolin-4-ol, substituted 1,3,4-oxadiazole derivative with 2-(5-thioxo-4,5-dihydro-1,3,4-oxadiazol-2-yl) phenyl acetate was done by the researcher18-20. Butalamine, Oxolamine, Furamizole, Pleconaril, ABT-751, and Fenadizole are the successfully used Oxadiazole nucleus containing marketed drug which shows various biological activity.21-25
Butalamine Oxolamine Furamizole:
Pleconaril ABT-751 Fenadizole
Figure 1.Oxadiazole nucleus containing marketed drug
There has been a considerable focus on conversion of Oxadiazole nucleus into trizole nucleus because of their remarkable biological property. The goal of the work, is to develop new amino acid substituted 1,2,4-trizole derivative from 1,3,4-oxadiazole nucleus.
MATERIALS AND METHODS:
All materials used were pharmacopoeial grade. Isoniazide hydrazide, 3-chloro benzyldehyde, Dimethyleformamide, Choloramine T, Glycine, and Luecine were from Central drug house, New Delhi. And other chemicals like Ethanol and methanol were from Hemedia Pvt. Ltd. Mumbai, India. The conformation of the synthesis was checked by thin layer chromatography using solvents and characterization was done by IR spectroscopy.
Scheme of work:
Step 1: General procedure for synthesis of Intermediate I:-
Isoniazide (0.01mol) and 3-chlorobenzeldehyde (0.01 mol) was transferred in a dry and clean conical flask. Add 5 drops of dimethylformamide (DMF) into the flask, mix it properly. Then the mixture goes for heating in commercial microwave oven at 300 W internally at 30 second for 3 minute. After 3 minute content of the flask was cooled with cold water, filtered and reaction content (intermediate Ⅰ) were collected. The reaction content then undergoes for washing, dried and recrystallization were done through ethanol (Scheme 1).
Step 2: Preparation of (2-(3-chloro benzene)-5-(4-pyridyl)-1, 3, 4-Oxadiazole:-
In a clean conical flask intermediate Ⅰ (0.01 mol) was mixed with ethanol (15ml).In the content chloramine-T (0.01 mol) was mixed. Then the content goes for heating in commercial microwave oven at 300 W internally at 30 second for 4 minute. Then the content of flask was cooled with cold water, filtered and reaction content (intermediate Ⅰ) was collected. The solid product (2-(3-chloro benzene)-5-(4-pyridyl)-1, 3, 4-oxadiazole were undergoes for washing, dried and recrystallization were done through methanol (Scheme 2).26
Step 3: Synthesis of 4-(4’-methyl pentanoic acid)-3(4-pyridyl)-5-(3-chloro benzene)-1,2,4-triazole (Compound A1):-
A mixture of 2-(3-chloro benzene)-5-(4-pyridyl)-1, 3, 4-oxadiazole (0.01 mol) and luecine (0.01 mol) in a dry and clean conical flask, was subjected for green synthesis in commercial microwave at 2450 MHz for 3 minute. Then the content of flask was cooled, filtered and reaction content was collected which further recrystallized from ethanol (Scheme 3).
Step 4: Synthesis of 4-(1’-ethanoic acid)-3(4-pyridyl)-5-(3-chloro benzene)-1,2,4-triazole (Compound A2):-
A mixture of 2-(3-chloro benzene)-5-(4-pyridyl)-1, 3, 4-oxadiazole (0.01 mol) and glycine (0.01 mol) in a dry and clean conical flask, was subjected for green synthesis in commercial microwave at 2450 MHz for 3 minute. Then the content of flask was cooled, filtered and reaction content was collected which further recrystallized from ethanol (Scheme 4).27
|
Scheme 1: Synthesis of intermediate Ⅰ (Step 1) |
|
Scheme 2: Synthesis of 2-(3-chloro benzene)-5-(4-pyridyl)-1, 3, 4-oxadiazole(Step 2) |
|
Scheme 4: Synthesis of compound A1(Step 3) |
|
Scheme 4: Synthesis of compound A1(Step 4) |
4-(4’-methyl pentanoic acid)-3(4-pyridyl)-5-(3-chloro benzene)-1,2,4-triazole (Compound A1):
Yield 82%, Rf value-0.89, m.p. 220-227 oC; FTIR (KBr) vmax cm-1: 3051.11 ( strtch Ar CH.), 1569.11 (strtch C=C.), 1301.20 (strtch C-C.), 3395.03 (strtch N-H.), 1125.01 (strtch N-N.), 1327.16 (strtch C-N.), 1586.13 (strtch C=N.); 1H NMR ,DMSO-d6 : 1.014-1.129 (d, 4H, C-H of aromatic pyridyl), 4.019-4.198 (m, 11H, aromatic benzyl C-H), 6.126-6.892 (t, 4H, Ar-H).
4-(1’-ethanoic acid)-3(4-pyridyl)-5-(3-chloro benzene)-1,2,4-triazole (Compound A2):
Yield 78%, Rf value-0.76, m.p. 232-240oC ; FTIR (KBr) vmax cm-1: 3051.18 ( strtch Ar CH.), 1569.02 (strtch C=C.), 1301.13 (strtch C-C.), 3395.06 (strtch N-H.), 1125.11 (strtch N-N.), 1327.25 (strtch C-N.), 1529.13 (strtch C=N.); 1H NMR DMSO-d6: 1.098-1.101 (d, 4H, C-H of aromatic pyridyl), 4.123-4.196 (m, 11H, aromatic benzyl C-H), 6.108-6.116 (t, 4H, Ar-H).
Antibacterial property:
Disc diffusion method was used to measure bacterial static charecterstics of substance. Inoculation of agar plates with standardized inoculums of the Escherichia Coli (gram -ve) and Staphylococcus aureus (gram +ve) was prepared then on the surface of the agar plate, filter paper discs were placed (about 6mm in diameter), which contain the test compound at a 0.1mg/ml concentration. The bacteria was inoculated on the drug plates with various amount of compounds. The agar plates were incubated at 370C for 24 hrs, and the presence or lack of bacterial growth in various plates was detected after 24 hrs. The compounds zone of inhibition was assessed against bacterial strains using ciprofloxacin as a reference drug.
Antioxidant property:
Antioxidant investigations were conducted using in vitro methods. 4.8mg of DPPH (1, 1-Diphenyl-2-picrylhydrazyl) was diluted with 20ml methanol, and the test tubes were covered with aluminium foil to shield it from light. For the control reading, 0.1ml of DPPH solution was mixed to 3ml of methanol and the absorbance was measured at 517nm. Different quantities of the chemical under investigation, as well as a standard compound (ascorbic acid), were obtained, and the volume was consistently created using methanol. Each sample was diluted in methanol to a final concentration of 3ml before adding 0.1ml of DPPH. On a UV-visible spectrometer, absorbance was measured after 15 minutes at 517nm using methanol as a blank. The following formula was used to compute the IC50 for each chemical as well as the standard preparation. The formula was used to calculate the DPPH free radical scavenging activity.27
% Scavenging = [absorbance of control – absorbance of test sample/absorbance of control] X 100
RESULTS AND DISCUSSION:
The synthesis of new amino acid substituted 1,2,4-triazole derivatives was carried out. Synthesis starts from the microwave assisted reaction between Isoniazide hydrazide and 3-chlorobenzeldehyde which yield intermediate compound and then with treatment with chloramines-T convert into Oxadiazole nucleus. In final step substitution of different amino acids lead the synthesis of new amino acid substituted 1,2,4-triazole derivatives.
Antibacterial characteristics of the compounds were analysed using the disc diffusion method with ciprofloxacin as a control medication, and the screening results of the synthesized compound are shown in (Table 1). Bacterial inoculation was done in agar plate and finally zone of inhibition was measured.
Table 1.Screening result of synthesized compound as antibacterial activity (mm)
|
S. No. |
Synthesized Compound |
Gram Positive Bacteria (S.Aureus) |
Gram Nagative Bacteria (E.Coli) |
Standard (Ciprofloxacin) |
|
1. |
A1 |
22 |
13 |
30 |
|
2. |
A2 |
20 |
11 |
30 |
Antioxidant activity was done by DPPH methods in which ascorbic acid were taken as standard drug. The UV absorbance in different concentration of synthesized compound and standard drug was measured (Table 2). And then Free radical inhibition activity (% inhibition) of synthesized compound and ascorbic acid was calculated using appropriate formula (Table 3).
Table 2.Absorbance of synthesized compound and ascorbic acid as antioxidant activity
|
S. No. |
Concentration (mg/ml) |
A1 |
A2 |
Standard (Ascorbic Acid) |
Control |
|
1. |
0.04 |
0.293 |
0.345 |
0.147 |
0.489 |
|
2. |
0.08 |
0.207 |
0.279 |
0.112 |
0.489 |
|
3. |
0.12 |
0.189 |
0.224 |
0.084 |
0.489 |
|
4. |
0.16 |
0.160 |
0.196 |
0.039 |
0.489 |
|
5. |
0.20 |
0.108 |
0.131 |
0.017 |
0.489 |
Table 3: Free radical inhibition activity (% inhibition) of synthesized compound and ascorbic acid
|
S. NO.
|
Concentration (mg/ml) |
A1 |
A2 |
Standard (Ascorbic Acid) |
|
1. |
0.04 |
40.08 |
29.45 |
69.94 |
|
2. |
0.08 |
57.67 |
42.94 |
77.09 |
|
3. |
0.12 |
61.35 |
54.19 |
82.82 |
|
4. |
0.16 |
67.28 |
59.92 |
92.02 |
|
5. |
0.20 |
77.91 |
73.21 |
96.52 |
CONCLUSION:
Compounds A1 and A2, which were synthesized by substituting leucine and glycine on the 1,2,4-triazole, were tested for antibacterial and antioxidant properties in vitro. The antibacterial activity was determined by measuring the zone of inhibition in mm against E. coli and S. aureus using the agar disc diffusion method. The antioxidant properties of synthesized compounds were measured using the DPPH radical scavenging method using ascorbic acid as standard medication for antibacterial activity. Solubility, pH melting point, optical rotation, TLC, UV, IR, antibacterial activity, and antioxidant activity were all tested on the produced compounds.
In comparison to Compound A2, the synthesized compound A1 had the highest MIC against S. aureus. Compound A1 also has the highest antioxidant activity. By observing this result it is divulged that a heterocyclic nucleus as triazole can have antimicrobial activity.
ACKNOWLEDGEMENT:
Mr. Jitendra Joshi, Chancellor and Professor Dr. Dharmbuddi, Vice chancellor, Research and Development Center Uttaranchal University, deserve particular gratitude for encouraging and giving research facilities to publish this research work.
CONFLICT OF INTEREST:
Nil.
REFERENCES:
1. Kumar AK. Lokeshwari DM. Pavithra G. Kumar VG. 1,2,4-Oxadiazoles: A potential pharmacological agents-An overview. Research Journal of Pharmacy and Technology. 2012; 5(12): 1490-1496.
2. Jacob J. Microwave assisted reaction in organic chemistry: A Review of recent advance. International Journal of Chemistry. 2012; 4: 6.doi.org/10.5539/ijc.v4n6p29
3. Ravichandran S. Microwave Synthesis- A Potential Tool for Green Chemistry. International Journal of Chem Tech Research. 2011; 3: 466-470.
4. Dallinger D. Kappe CO. Microwave-Assisted Synthesis in Water as Solvent. American Chemical Society. 2007; 107: 2563−2591.doi.org/10.1021/cr0519410
5. Mohamed MO. Ball Milling for Heterocyclic Compounds Synthesis in Green Chemistry: A Review. Synthetic Communications. 2015; 1532-2432.doi.org/10.1080/00397911.2015.1058396
6. Robert EB. Review of Heterocyclic Chemistry. 2012; 8: 1349–1350.
7. Patel KJ. Review of synthesis of 1,3,4-Oxadiazole derivatives. Synthetic Communications. 2014; 44: 1859–1875.doi.org/10.1080/00397911.2013.879901
8. SalahuddiN. Mazumder A. Updates on synthesis and biological activities of 1,3,4- oxadiazole: A review. Synthetic Communications. 2017; 47: 1805-1847. doi.org/10.1080/00397911.2017.1360911
9. Khudhair ZT. Al-Tamimi EO. Synthesis, Identification, Theoretical Study and effect of 1,3,4-Oxadiazole Compounds Substituted on Creatinine ring on the activity of some Transfers Enzymes. Research Journal of Pharmacy and Technology. 2019; 12(8):3581-3588. Doi.org/10.5958/0974-360X.2019.00611.5
10. Az-Eddine ME. Design, synthesis, chemical characterization, biological evaluation, and docking study of new 1,3,4-oxadiazole homonucleoside analogs. Nucleosides, Nucleotides & Nucleic Acid. 2020; 1525. DOI:10.1080/15257770.2020.1761982
11. Szafrański K. Synthesis, anticancer evaluation and structure-activity analysis of novel (E)-5-(2-arylvinyl)-1,3,4-oxadiazol-2-yl)benzenesulfonamides. International Journal of Molecular Science. 2020; 21: 1–23. DOI: 10.3390/ijms21062235
12. Kakanejadifard A. The synthesis, structural characterization and antibacterial propertiesof some 2-((4-amino-1,2,5-oxadiazol-3-ylimino)methyl)-4- (phenyldiazenyl)phenol. Dyes and Pigments. 2013; 97: 215-221. DOI:10.1016/j.dyepig.2012.10.004
13. Kumar R. Shahar MY. Rai AK. Chaturvedi S. Synthesis and biological evaluation of some novel 1,3,4-oxadiazoles derived from bi phenyl 4- carboxylic acid. Der Pharmacia Lettre. 2013; 5: 366-370.
14. Shridhar AH. Synthesis of 1,3,4-oxadiazole incorporated azo dye derivatives as a potent biological activity. Molecules. 2012; 4: 119-129.
15. Cai ZW. Wei D. Borzilleri RM. Qian L. Kamantg A. Mortillo S. Waulet B et al Synthesis, SAR, and evaluation of 4-[2,4-difluoro-5- (cyclopropylcarbamoyl)phenylamino]pyrrolo[2,1-f][1,2,4]triazinebased VEGFR-2 kinase inhibitors. Bioorganic and Medicinal Chemistry. 2008; 18: 1354. DOI: 10.1016/j.bmcl.2008.01.012
16. Almajan GL. Barbuceanu SF. Innocenti A. Scozzafava A. Supuran CT. Carbonic anhydrase inhibitors. Inhibition of the cytosolic and tumor-associated carbonic anhydrase isozymes I, II and IX with some 1,3,4-oxadiazole- and 1,2,4-triazole-thiols. Journal of Enzyme Inhibition and Medicinal Chemistry. 2008; 23: 101-107. doi.org/10.1080/14756360701342417
17. Zadorozhnii PV. Kiselev VV. Teslenko NO. Kharchenko AV. Pokotylo IO. Okhtina OV et al In Silico Prediction and Molecular Docking Studies of N-Amidoalkylated Derivatives of 1,3,4-Oxadiazole as COX-1 and COX-2 Potential Inhibitors. Research Journal of Pharmacy and Technology. 2017; 10(11): 3957-3963. Doi.org/10.5958/0974-360X.2017.00718.1
18. Muhammed KM. Sulaiman AH. Nihad IT, Preparation and Identification of some new Compounds 1,3,4-Oxadiazole derivatives using Grinding Technique. Research Journal of Pharmacy and Technology. 2018; 11(10): 4272-4276. doi.org/10.5958/0974-360X.2018.00783.7
19. Unnissa SH. Karthikaa T. Kavitha K. Kishore PS. Kowsalya M. Synthesis and Biological Evaluation of some 2,5-disubstituted 1,3,4-oxadiazole based Cinnoline derivatives. Research Journal of Pharmacy and Technology. 2019; 12(10): 4812-4816. Doi.org/10.5958/0974-360X.2019.00832.1
20. Singhai A. Gupta MK. Synthesis and Characterization of 1,3,4-Oxadiazole Derivatives as Potential Anti-inflammatory and Analgesic agents. Research Journal of Pharmacy and Technology. 2020; 13(12):5898-5902. Doi.org/10.5958/0974-360X.2020.01029.X
21. Garewal H. Review on synthesis and spectral analysis of 1,3,4-oxa/thiadiazole moiety based on their literature support. World Journal of Pharmaceutical Research. 2019; 8: 285-306.
22. Murty MSR. Penthala R. Nath LR. Anto RJ. Synthesis of Salicylic Acid-based 1,3,4-oxadiazole Derivatives Coupled with Chiral Oxazolidinones: Novel Hybrid Heterocycles as Antitumor Agents. Letters in Drug Design & Discovery. 2014; 11: 1133-1142. DOI: 10.2174/1570180811666140627004607
23. Patel KD. Vekariya RH. Patel HD. A One Pot, Solvent Free and Catalyst Free Synthesis of Substituted 2- Amino-5-Aryl-1,3,4-Oxadiazoles under Microwave Irradiation. Current Microwave Chemistry. 2016; 3. DOI: 10.2174/2213335602666150324233632
24. Kharb R. Kaur R. Sharma AK. Vistas on antimicrobial potential of novel Oxadiazole derivatives in modern medicinal chemistry, European Journal of Biomedical and Pharmaceutical Sciences. 2014; 1.
25. Tiwari D. Narang R. Khokra SL. A review on microwave assisted synthesis, mechanism of action and structure activity relationship of 1, 3, 4-oxadiazole derivatives as anticancer agent. World Journal of Advanced Research and Reviews. 2020; 9: 086–096.doi.org/10.3057/wjarr.2021.9.1.0472
26. Biju CR. Ilango K. Prathp M. Rekha K. Design and Microwave-assisted synthesis of 1,3,4-Oxadiazole Derivative for analgesic and anti-inflammatory activity. Journal of Young Pharmacists. 2012; 4. doi: 10.4103/0975-1483.93576
27. Joshi H. Bisht AS. Juyal D. Synthesis and characterization of Triazole compound derived from Oxadiazole moiety for its antimicrobial and anti-oxidant evaluation. Internation journal of Medicinal Chemistry and Analysis. 2020; 10: 22-30.doi.org/10.21276/ijmca.2020.10.1.4
Received on 04.02.2022 Modified on 29.04.2022
Accepted on 18.07.2022 © RJPT All right reserved
Research J. Pharm. and Tech 2023; 16(5):2322-2326.
DOI: 10.52711/0974-360X.2023.00382