Novel Synthesis, Characterization and Biological Evaluation of Substituted Pyrazolidine-3, 5-Dione
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Central JSM Chemistry Research Article *Corresponding author Neeraj Upmanyu, School of Pharmacy & Research, By-Pass Road, Bhanpur, Bhopal (M.P.) -462027, Tel: 91- Novel Synthesis, 9425-128642; Email id: Submitted: 06 July 2015 Characterization and Biological Accepted: 31 August 2015 Published: 02 Spetmeber 2015 ISSN: 2333-6633 Evaluation of Substituted Copyright © 2015 Upmanyu et al. Pyrazolidine-3, 5-Dione OPEN ACCESS 1 1,2 1 Rohit Singh , Neeraj Upmanyu *, Manish Gupta and Pallavi Keywords Prasad3 • Pyrazolidine-3, 5-dione 1Department of Pharmaceutical Chemistry, R.K.D.F. College of Pharmacy, India • Benzoic acid 2School of Pharmacy and Research, Peoples University, India • Antibacterial & Anti-inflammatory 3School of Pharmaceutical Sciences, India Abstract An efficient lead molecule Pyrazolidine-3, 5-dione derivative has been developed as anti-inflammatory and antibacterial agents. In the present investigation, the compounds (RS-1 to RS-10) were synthesized according to Scheme 1 using 4-substituted benzoic acid as starting material. This acid is converted into the respective ester (R1 to R5) and then ester is converted into hydrazide (R2A1 to R2A5 & R2B1 to R2B5), and finally into the pyrazolidine-3,5-diones (tested compound). The compounds were evaluated for their anti-inflammatory activity by the carrageen an-induced paw edema method and anti-bacterial activity using zone of inhibition method. Out of these compounds RS-6, RS-9, RS-10, and RS-2 were found to be the most active synthesized compounds, with significant anti-inflammatory activity. The Structure Activity Relationship (SAR) of these synthesized compounds showed that substitution with Nitro and Chloro group at para position of phenyl ring is produce optimum activity, as among all the synthesized compounds, the most active ones were RS-6, RS-9, and RS-10. Another structural feature of all the synthesized compounds was that phenyl substitution over Nitrogen of Pyarazolidine-3,5-dione also produce appreciable activity. Therefore these synthesized molecules can be considered as lead molecules for future investigations. ABBREVIATIONS on heterocycles and the chemistry of Pyrazolidine-3,5-dione has received considerable attention owing to their synthetic and FAB: Fast Atom Bombardment; TLC: Thin Layer effective biological importance. Pyrazolidine-3,5-dione moiety Chromatography; IR: Infra Red; OECD: Organization for Economic has been incorporated into a wide variety of therapeutically Cooperation and Development INTRODUCTION antimicrobial [1]. There are many marketed drugs containing the interesting drug candidates including anti-inflammatory and In the last two decades, the chemistry of pyrazolidine-3,5- [3], Phenyl butazone, Kebuzone, Mofebutazone [4] etc. From dione and their fused heterocyclic derivatives have received thepyrazolidine-3,5-diones literatures, it may group be predictede.g., Feprazone that [2], Pyrazolidine-3,5- Sulfinpyrazone considerable attention owing to their synthetic and effective dione moiety represents important pharmacophore and plays biological importance. In spite of a large number of antibiotics a vital role in medicinal agents. A degree of respectability has and chemotherapeutics available for medicinal use, the treatment been bestowed upon Pyrazolidine-3,5-dione derivatives due of infectious diseases still remains an important and challenging to their wide range of biological activities such as antibacterial problem because of a combination of factors including emerging infectious diseases and the increasing number of multi-drug hypoglycemic properties [9]. In the design of new drugs, the resistant microbial pathogens, so it revealed a substantial need combination[5], antifungal of [6],different anti-inflammatory pharmacophores [7], frameanalgesic may [8], lead and to for new classes of antimicrobial agents. There is really perceived need for the discovery of new compounds endowed with different phenyl incorporated Pyrazolidine-3,5-dione displayed varied activity. Through the various molecules designed and synthesized pharmacologicalcompounds with interestingproperties. biologicalPrompted profiles. by these As investigations alkyl, benzyl, for this aim, in recent years, active research has been initiated we synthesized compounds containing Pyrazolidine-3,5-dione Cite this article: Singh R, Upmanyu N, Gupta M, Prasad P (2015) Novel Synthesis, Characterization and Biological Evaluation of Substituted Pyrazolidine-3, 5-Dione. JSM Chem 3(1): 1016. Upmanyu et al. (2015) Email: Central with attached benzoyl group and evaluated them for their anti- R1 OH R1 O CH3OH MATERIALS AND METHODS inflammatory and antimicrobial activity [10]. Conc. H2SO4 O O Chemicals P-Substituted Benzoic Acid Methyl Ester Of Substituted Benzoic Acid All the solvents were of LR grade and were obtained from R1 -R5 Merck, Rankem, Fluka, and S.D. Fine Chemicals. The Melting O C2H5OH R2A/R2B points were recorded in open capillary tubes and are uncorrected. O O O O H using silica gel G. Visualization was done using iodine vapours or N Diethyl Malonate N R2 The purity of the synthesized compounds was confirmed by TLC H R1 sulfuric acid (30% v/v). spectrophotometer. Chemical shifts were expressed in parts Substituted Hydrazide O IR spectra were 13recorded in KBr discs in Nicolet-6700 R2A1 -R2A5 C NMR spectra were recorded on Bruker & R as the solvent, 2 R2B1 -R2B5 6 N N per million (δ) units. HERE, R = NO /OH/Cl/H/NO TMS as an internal standard. Mass spectra were recorded on R1 1 2 2 Avance II 400 MHz apparatus using DMSO-d R2 = H / Ph O O R2A = Ph-NH.NH2 technique and nitrogen analysis was done on elemental analyzer R2B = NH2.NH2.H2O Jeol Sx 102/DA-600 mass spectrometer/Data system using FAB RS-1 - RS-10 Figure 2 Scheme of Synthesis of Title Compound. Elementar Vario EL III Carlo Erba 1108. the Anti-inflammatoryexperimental protocol and wasanalgesic approved activities by the were Animal performed Ethical (Step-1) Synthesis of Different Para Substituted in the animal house of the R.K.D.F College of Pharmacy (M.P.), and Benzoic Acid Methyl Esters (R1 to R5) Committee of R.K.D.F College of Pharmacy (M.P.)-India. (780/05/ ab/CPCSEA).Animals and microbial cultures One mole of substituted benzoic acid was dissolved in 70- chips80 mL were methanol putted and to avoid taken bumping. in a round The bottom whole flask. reaction After mixture it 1.5- 2.0ml sulphuric acid was added from side and some porecelain laboratoryWister albinoconditions rats (120-200 maintained g) and at swiss25±1° Calbino and micefewer (20-30 than cooled to room temperature and equal quantity of water has been g) of either sex were used. The animals housed under standard addedwas subjected to it. This to makesreflux forester 6hrs. and After water refluxing layer separated the mixture but was the two layer was not distinctive one, so for this ester is separated Animal12 / 12 experiments h light /dark were cycle approved and fed by with the standardInstitutional pellet Animal diet with the help of carbon tetrachloride from separating funnel and Ethical(Gold Mohur Committee brand, for Lipton the purpose India Ltd.) of Control and water and Supervisionad libitum. the funnel, removed the sodium bicarbonate layer and tested to seewashed if it waswith basic.0.6M sodiumSeparated bicarbonate. the organic After layer shaking and dried and venting it with the directives of Ministry of Social Justice and Empowerment, of Experiments on Animals (CPCSEA-780), constituted under anhydrous magnesium sulphate & removed the MgSO by gravity Government of India. The microbial cultures of bacteria Proteus 4 mirabilis Pseudomonas aeruginosa ester was obtained which is again recrystallized with ethanol. Bacillus subtilis Staphylococus aureus filtration. Carbon tetrachloride was evaporated and respective were used (MTCC for the 425),evaluation of anti-bacterial activity. (MTCC 424), (MTCC 619) and (MTCC 96) Yield(Step is 89%.- 2) Synthesis of Different substituted Chemistry Hydrazides of Substituted Methyl Benzoates (R2A1 to The preparation of different substituted pyrazolidine-3,5- R2A5 and R2B1 to R2B5) dione by the initial reagent i.e, from substituted benzoic acid consist of three steps [11,12]. Synthesis of the Pyrazolidine-3,5- 0.1mole of different Substituted methyl benzoates previously dissolved in ethanol was taken in round bottom flask and mixed dione derivatives is shown in (figure 2). drops of acetic acid was also added along with some porcelain with 9-10ml of hydrazine hydrate or phenyl hydrazine, 1-2 O R2 chips. The whole system is subjected to reflux for 8-9 hrs. After N N refluxing product (respective hydrazide) was obtained which is O again(Step-3) recrystallize Synthesis with of absolute Different ethanol? Substituted Yield is 73%. 1-benzoyl- pyrazolidine-3,5-dione (RS-1 to RS-10) R 1 O After the successful completion of the above two steps the Figure 1 Basic Molecule. last step concludes the reaction with cyclisation, the formation JSM Chem 3(1): 1016 (2015) 2/7 Upmanyu et al. (2015) Email: Central of different substituted pyrazolidine-3,5-dione molecule group received an equivalent amount of CMC. To one group the was carried out. For this equimolar quantity of the different substituted hydrazide was reacted with diethyl malonate to wasstandard injected drug into Indomethacin the sub plantar (10 mg/kg) tissue ofwas the administered. left paw of all After the one hour, carrageen an (0.1 mL, 1% w/v solution in sterile saline) obtainBiological the final Evaluation product. Yield is 41%. In biological evaluation antibacterial activity and anti- animals. The right paw served as the reference non-inflamed paw for comparison [18]. The initial paw volume was measured using percentagea plethysmograph of reduction within in30 theseconds paw ofvolume the injection. was calculated After three by inflammatoryAnti-Bacterial activity Activity was performed. subtractinghours, the final the pawdifference volume between of each theanimal right was and measured. left hind pawThe The newly synthesized compounds were screened for volumes in the treated group from the difference in the control their antibacterial activity against bacterial strains by disc group and dividing it by the difference in the control group.