Research Article Anti-Inflammatory, Analgesic Evaluation...

Hindawi Publishing CorporationJournal of ChemistryVolume 2013 Article ID 412053 6 pageshttpdxdoiorg1011552013412053

Research ArticleAnti-Inflammatory Analgesic Evaluation andMolecular Docking Studies of N-Phenyl AnthranilicAcid-Based 134-Oxadiazole Analogues

Suman Bala1 Sunil Kamboj1 Vipin Saini1 and D N Prasad2

1 M M College of Pharmacy Maharishi Markandeshwar University Mullana Ambala Haryana 133207 India2 Shivalik College of Pharmacy Nangal Ropar Punjab 140126 India

Correspondence should be addressed to Suman Bala sumankmj7gmailcom

Received 31 May 2013 Accepted 21 August 2013

Academic Editor Frank Wuest

Copyright copy 2013 Suman Bala et al This is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

A novel series of N-phenyl anthranilic acid-based 134-oxadiazoles were prepared (4andashh) and subjected to anti-inflammatoryanalgesic activity andmolecular docking studies to target cyclooxygenase-2 enzyme 134-Oxadiazole derivatives were screened foranti-inflammatory activity in carrageenan-induced rat paw edema and analgesic activity by tail immersion method In synthesizedcompounds the free carboxylic group which is responsible for gastric side effects was derivatized by heterocyclic 134-oxadiazolebioactive core which showed good interaction with COX-2 receptor with good docking score Among all the synthesizedcompounds 4e and 4f have emerged out as potential COX-2 inhibitors

1 Introduction

Non-steroidal anti-inflammatory drugs (NSAIDs) form animportant class of widely used therapeutic agents due totheir anti-inflammatory analgesic and antipyretic effectsThe pharmacological activity of NSAIDs is related to sup-pression of prostaglandin biosynthesis by inhibiting theenzyme cyclooxygenase (COX) COX is an endogenousenzyme which catalyzes the conversion of arachidonic acidinto prostaglandins and thromboxanes The enzyme existsin at least two isoforms COX-1 and COX-2 Althoughboth isoforms catalyze the same biochemical transforma-tion they are subject to a different expression regulationCOX-1 is a constitutive enzyme and is responsible for thephysiological function of prostaglandins (PGs) like main-tenance of the integrity of the gastric mucosa and pro-vides adequate vascular homeostasis whereas COX-2 is aninducible enzyme and is expressed only after an inflammatorystimulus [1]

With the chronic use of NSAIDs one prominentside effect is the formation of gastric ulcers Heterocycliccompounds containing five-membered oxadiazole nucleus

possess a diversity of useful biological effects such asantiedema and anti-inflammatory activities 134-Oxadi-azoles have anti-inflammatory activity by virtue of dualmechanism that is inhibiting both COXLOs to reducegastric ulcer formation [2] Literature studies suggest thatdirect tissue contact of NSAIDs plays an important role inthe production of side effects like gastric upset irritationand ulceration [3 4] and the reported literature confirmsthat gastrointestinal side effects of NSAIDs such as irritationand GI bleeding are due to the presence of a free carboxylicgroup in the parent drug [5 6] Thus developing new agentswithminimumorwithout side effects is an extensive researcharea in the present scenario Our studies and studies of otherresearchers [6] have shown that derivatization of the car-boxylate function of some NSAIDs resulted in an increasedanti-inflammatory activity with a reduced ulcerogenic effectHence it is not irrelevant to speculate that replacing theterminal carboxylic function of NSAIDs by oxadiazole ringa five membered heterocyclic nucleus may enhance theanti-inflammatory activity of such compounds Hence byincorporating the oxadiazolyl moiety we hope to get a betteranti-inflammatory molecule

2 Journal of Chemistry

NH OO

NN

R

4a R =

4b NH2R =

OH4c R =

OH

4d R =

Cl4e R =

Cl

4f R =

4g CH3R =

4h NO2R =

4andashh

Figure 1 N-Phenyl anthranilic acid-based substituted 134-oxadiazole analogues (4andashh)

2 Material and Method

21 Preparation of Target Compounds Target compounds(4andashh) were prepared previously in our laboratory [7](Figure 1)The aromatic esterswere prepared by Fischer ester-ification Further these esters were subjected to formationof aryl hydrazide in presence of hydrazine hydrate Arylhydrazide 2a (1M)was dissolved in phosphorous oxychloride(5mL) and to it compound 3 (equimolar 1M) was addedThe reaction mixture after refluxing for 6-7 h was cooledto room temperature and poured onto crushed ice On neu-tralization of the contents with sodium bicarbonate solution(20) a solid mass separated out It was filtered and washedwith water It was crystallized by using methanol to give4a Similarly compounds 4bndashh were prepared The physicalproperties and spectral characterization were already dis-cussed [7]

22 Chemicals and Instruments All chemicals used in theestimation were of analytical grade Carrageenan was pur-chased from Sigma Chemicals A digital plethysmometer(model pth-7070 srnopt 070509 Medicad system) was usedfor measuring paw volume (mL) of rats edema Molecu-lar docking studies were carried out on Molecular VirtualDocker 500

23 Animals Albino rats of either sex around 180ndash200 gof weight were selected for both anti-inflammatory andanalgesic activity respectively The animals were kept inpolypropylene cages (3 in each cage) at an ambient tem-perature of 20 plusmn 2∘C and 55ndash65 relative humidity A 12-12 hrs light and dark schedule was maintained in the animalhouse The rats had free access to water and food Theexperimental protocol was approved by the InstitutionalAnimals Ethical Committee (MMCPIEC1016) and animalcare was done as per the guidelines of the committee forthe purpose of control and supervision of experiments onanimals (CPCSEA) Government of India

24 Acute Toxicity Studies It has been fund from the lit-erature survey that 134-oxadiazole derivatives in a dose of1000mgkg produce mortality (LD

50) in rats Soon tenth of

the LD50 that is 100mgkg (ED

50) was selected as a dose for

anti-inflammatory and analgesic activity [8]

25 In Vivo Anti-Inflammatory Activity In carrageenanmodel albino rats of all groups were treated with subcuta-neous injection of 01mL of 1 wv solution of carrageenaninto the subplantar region of the right hind paw The pawwas marked with permanent marker at the planter regionwhere the paw volume was to be measured The diclofenacsodium (10mgkg) and test compounds (100mgkg) weresuspended in 03 sodium carboxy methyl celluloseThe testcompounds and vehicle (control) were administered po withthe help of gastric cannula half an hour after the injection ofcarrageenan in subplanter region of right paw Mean normalpaw volume was measured 30min prior to carrageenaninjection by using plethysmometer Mean increase in the pawvolume for control group (after carrageenan injection) andtest group was measured at 1 hr 2 hr and 3 hr [9] Percentinhibition of inflammation after teststandard was calculatedusing the formula

inhibition =119881119888minus 119881119905

119881119888

times 100 (1)

where 119881119905is the of paw volume (mL) of teststandard com-

pound at corresponding time and 119881119888is the paw volume (mL)

of control

26 Analgesic Activity Tail immersion method is based onthe observation that morphine-like drugs selectively prolongthe reaction time of the typical tail withdrawal reflex inmice Albino rats were divided in twenty-six groups eachcontaining six animals The tail of mice was immersed (1-2 cm) in warm water kept constant at 55∘CThe reaction timewas recorded by stopwatch (the reaction time is the timetaken by the rats to flick their tails) The latent period of thetail flick response will be determined before and 15 30 60and 120min after drug administration [10]

27 Statistical Analysis The results were expressed as mean plusmnSEM for six animals in each group for anti-inflammatory andanalgesic activity All the grouped data was statistically evalu-atedHypothesis testingmethod included one-way analysis of

Journal of Chemistry 3

Table 1 In vivo anti-inflammatory activity of synthesized 134-oxadiazole analogues

Change in paw edema volume (mL)Groups 1 h Percentage inhibition 2 h Percentage inhibition 3 h Percentage inhibitionControl 043 plusmn 005 mdash 072 plusmn 006 mdash 098 plusmn 009 mdashDiclofenac sodium (10mgkg) 025 plusmn 003 5713 030 plusmn 002 5833 026 plusmn 001 73464a (100mgkg) 040 plusmn 005 697 061 plusmn 008 1528 069 plusmn 006 29594b (100mgkg) 039 plusmn 001 930 060 plusmn 006 1666 059 plusmn 004 39794c (100mgkg) 038 plusmn 005 116 048 plusmn 004 3334 045 plusmn 003 54084d (100mgkg) 036 plusmn 001 1627 052 plusmn 003 2778 049 plusmn 004 504e (100mgkg) 029 plusmn 002

lowast 3255 038 plusmn 004lowast 4723 031 plusmn 002

lowast 68364f (100mgkg) 034 plusmn 005 209 043 plusmn 003

lowast 4028 036 plusmn 006lowast 6326

4g (100mgkg) 036 plusmn 005 1627 055 plusmn 002 2361 055 plusmn 005 43874h (100mgkg) 035 plusmn 001 186 046 plusmn 002 3612 044 plusmn 003 5310All values are expressed as means plusmn SEM (119899 = 6) lowast119875 lt 005 versus control

variance (ANOVA) followed by Dunnettrsquos comparison test 119875values of less than 005 were considered to indicate statisticalsignificance

28 Molecular Docking Studies Comparative docking of aset of ligands with specific proteins involves methodologywith easy user interface and their respective scoring functionprovided by Molegro Virtual Docker Molecular dockingstudies were performed to target COX-2 The compoundswhich have shown potent anti-inflammatory and analgesicactivity (4e and 4f) were subjected to molecular docking totarget COX-2 (Pdb-1CX2) [11 12]Steps in methodology are as follows

(1) importing a protein file and ligand file and prepara-tion of ligands

(2) protein preparation and detecting cavities of proteinmolecules

(3) executing a docking setup through docking wizardpanel

(4) poses of protein-ligand complexes obtained afterdocking process with their specific mol dock scoresdisplayed in output file

3 Results

31 In Vivo Anti-Inflammatory Activity 134-Oxadiazolecompounds were evaluated for anti-inflammatory activityin carrageenan-induced rat paw edema Among all thecompounds 4e and 4f have shown maximum activity at100mgkg (Table 1) (Figure 2)

32 Analgesic Activity 134-Oxadiazole compounds wereevaluated for analgesic activity by tail immersion methodAmong all the compounds 4e and 4f have shown maximumanalgesic activity at 100mgkg (Table 2) (Figure 3)

33 Molecular Docking Studies 134-Oxadiazole compoundswere subjected to molecular docking to target COX-2 Com-pounds 4e and 4f have shown good interaction with COX-2 The interaction of ligands 4e and 4f diclofenac sodium

0

02

04

06

08

1

12

Redu

ctio

n in

paw

vol

ume (

mL)

Target compounds

Con

trol4a 4b 4c 4d 4e 4f 4g 4h

After 3hours

Dic

lofe

nac

sodi

um

Figure 2 This histogram shows the inhibition of carrageenan-induced paw edema in rats The histogram is plotted between targetcompounds (4andashh) at 100mgkg control standard drug diclofenacsodium (10mgkg) on X axis and reduction in paw volume on Yaxis Data has shown that the inhibition of paw edema significantlydecreases by target compounds from control Data are Mean plusmnSEM indicates significant decrease in inflammation form controland indicates highly significant decrease 119875 lt 005

0

5

10

15

20

25

Reac

tion

time (

s)

Target compounds

Con


Dic

lofe

nac

sodi

umAfter 120min

Figure 3 This histogram shows the reaction time in rats bytail immersion method The histogram is plotted between targetcompounds (4andashh) at 100mgkg control standard drug Diclofenacsodium (10mgkg) on X axis and reaction time (sec) on Yaxis Data showed that the reaction time significantly increases bytarget compounds from control Data are Mean plusmn SEM indicatessignificant increase in reaction time form control and indicateshighly significant increase 119875 lt 005


4e 4e 998400

4f 4f 998400

Figure 4 Binding modes of 4e 4f (4e 4f as docking view and 4e1015840 4f1015840 as interaction view) with COX-2 where bluegreen lines and red linesrepresent hydrogen bonding and favourable steric interactions respectively

Table 2 Analgesic activity of synthesized 134-oxadiazole analogues

Reaction time (sec)Groups (After 30min) (After 60min) (After 120min)Control 543 plusmn 17 529 plusmn 09 630 plusmn 12

Diclofenac sodium (10mgkg) 507 plusmn 16 1060 plusmn 20 1883 plusmn 24

4a (100mgkg) 417 plusmn 09 646 plusmn 002 942 plusmn 19

4b (100mgkg) 580 plusmn 14 607 plusmn 002 827 plusmn 24

4c (100mgkg) 583 plusmn 16 861 plusmn 008 1135 plusmn 16

4d (100mgkg) 442 plusmn 07 760 plusmn 006 1032 plusmn 20

4e (100mgkg) 587 plusmn 11 1034 plusmn 009lowast

1537 plusmn 20lowast

4f (100mgkg) 472 plusmn 10 957 plusmn 001lowast


4g (100mgkg) 567 plusmn 08 744 plusmn 005 1003 plusmn 25

4h (100mgkg) 456 plusmn 11 611 plusmn 008 930 plusmn 23

All values are expressed as means plusmn SEM (119899 = 6) lowast119875 lt 005 versus control

and selective cyclooxygenase inhibitor-558 with receptor interms of docking score (binding energy) was depicted inTable 3 (Figures 4 and 5)

4 Discussion

The common side effect associated with regular use ofnonsteroidal anti-inflammatory drugs is gastric ulcerationwhich is due to the presence of free acid functionality inthe structure of the molecule This acidic group was masked

by the introduction of heterocyclic 134-oxadiazole nucleusA series of 134-oxadiazole derivatives which were previ-ously synthesized in our laboratory were subjected to anti-inflammatory analgesic and molecular docking studies totarget cyclooxygenase-2 From all the compounds 4e and 4fhaving a chloro group at para and ortho positions were foundto be potent in both analgesic and anti-inflammatory activitywhich is comparable to standard Also the presence of paraand ortho substituted halogens (ndashCl) in the compound resultsin enhanced biological activities This significant increase


(SC-S58)

(Diclofenac) (Diclofenac998400)

(SC-S58998400)

Figure 5 Bindingmodes of diclofenac SC-S58 (diclofenac SC-S58 as docking view and diclofenac1015840 SC-S581015840 as interaction view)withCOX-2where bluegreen lines and red lines represent hydrogen bonding and favourable steric interactions respectively

Table 3 Ligand-receptor interaction of target compound and standard drugs

Compound Docking score Distance Amino acid Group involved

4e minus77760 283 Glu 87 ndashNHndash of N-phenyl anthranilic acid274 Glu 87 Carbonyl group of N-phenyl anthranilic acidic group

4f minus76499

306 Lys 492 ndashOndash of oxadiazole ring284 Lys 492 ndashNndash of oxadiazole313 Lys 492 ndashNndash of oxadiazole304 Glu 479 ndashNndash of N-phenyl group

Diclofenac minus58259 269 Lys 492 ndashOndash of nitro group

SC-S58 minus91779302 Ile 112 ndashNndash of SO2NH2 group271 Arg 120 ndashNndash of diazole ring328 Ser 119 ndashNndash of diazole ring

in biological activities is attributed due to the electronwithdrawing nature of halogens which ultimately resultsin enhancement in lipophilicity This enhanced lipophilicitycould facilitate the penetration or passage of these com-pounds across the biological membrane easily

The docking scores of 4e and 4f were much more thanthat of standard drug diclofenac sodium but less than thatof SC-S58 (selective COX-2 inhibitor) The docking score isrepresentative of binding energy of ligand to receptor Theamino acids that interacted with COX-2 in 4e are Glu 87and Lys 492 The replacement of free carboxylic group byoxadiazole nucleus in target compounds 4e and 4f enhanced

the interaction by formation of numerous hydrogen bondswith cyclooxygenase receptor These results could be usedfor the development of novel potent and effective COX-2inhibitors

The carbonyl 134-oxadiazole (act as acceptor) and ndashNHndash (act as donor) functionalities in the synthesized deriva-tives have played very important role in ligand-receptorinteraction for the formation of hydrogen bonds Results alsorevealed that the hydrogen bond distance is important indocking studies Distance more than 32 A indicates weakhydrogen bond bonding between ligand and receptor thedistance 26 Andash32 A represents a good hydrogen bonding


5 Conclusion

Derivatives having a 134-oxadiazole nucleus instead of a freecarboxylic group are devoid of gastrointestinal side effectsThe compounds 4e and 4f have shown good interaction withCOX-2 enzyme and thus emerged out as potential lead forthe development of novel anti-inflammatory agent with goodefficacy and lesser side effects

Acknowledgment

The authors are thankful to the M M College of Phar-macy Maharishi Markandeshwar University for providingthe facility to carry out the research

References

[1] A Palomer F Cabre J Pascual et al ldquoIdentification of novelcyclooxygenase-2 selective inhibitors using pharmacophoremodelsrdquo Journal of Medicinal Chemistry vol 45 no 7 pp 1402ndash1411 2002

[2] F A Omar N M Mahfouz and M A Rahman ldquoDesign syn-thesis and antiinflammatory activity of some 134-oxadiazolederivativesrdquo European Journal of Medicinal Chemistry vol 31no 10 pp 819ndash825 1996

[3] F L Lanza ldquoA guideline for the treatment and prevention ofNSAID-induced ulcersrdquo American Journal of Gastroenterologyvol 93 no 11 pp 2037ndash2046 1998

[4] F Buttgereit G R Burmester and L S Simon ldquoGastrointesti-nal toxic side effects of nonsteroidal anti-inflammatory drugsand cyclooxygenase-2-specific inhibitorsrdquo American Journal ofMedicine vol 110 no 3 pp 135ndash195 2001

[5] K A Metwally S H Yaseen E-S M Lashine H M El-Fayomi and M E El-Sadek ldquoNon-carboxylic analogues ofarylpropionic acids synthesis anti-inflammatory activity andulcerogenic potentialrdquo European Journal of Medicinal Chem-istry vol 42 no 2 pp 152ndash160 2007

[6] A Husain M S Y Khan S M Hasan and M M Alamldquo2-Arylidene-4-(4-phenoxy-phenyl)but-3-en-4-olides synthe-sis reactions and biological activityrdquo European Journal ofMedicinal Chemistry vol 40 no 12 pp 1394ndash1404 2005

[7] S Bala S Kamboj V Saini and D N Prasad ldquoSynthesischaracterization computational studies and evaluation of novelsubstituted 1 3 4-oxadiazolesrdquo International Journal of Phar-macy and Pharmaceutical Sciences vol 4 pp 131ndash135 2012

[8] Inderba Indian drugs ldquoIndian Drugs manufacturing associa-tion 102B Poonam chambersrdquo Dr A B Rd Worli BombayIndia 19301 1982

[9] C A Winter E A Risley and G W Nuss ldquoCarrageenin-induced edema in hind paw of the rat as an assay for anti-iflammatory drugsrdquo Proceedings of the Society for ExperimentalBiology and Medicine vol 111 pp 544ndash547 1962

[10] H G Vogel Drug Discovery and Evaluation PharmacologicalAssay vol 697 Springer New York 2nd edition 2002

[11] A Bali R Ohri and P K Deb ldquoSynthesis evaluation anddocking studies on 3-alkoxy-4-methanesulfonamido acetophe-none derivatives as non ulcerogenic anti-inflammatory agentsrdquoEuropean Journal of Medicinal Chemistry vol 49 pp 397ndash4052012

[12] M Lindner W Sippl and A A Radwan ldquoPharmacophoreelucidation and molecular docking studies on 5-phenyl-1-(3-pyridyl)-1H-124-triazole-3-carboxylic acid derivatives asCOX-2 inhibitorsrdquo Scientia Pharmaceutica vol 78 no 2 pp195ndash214 2010

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Spectroscopy

Analytical ChemistryInternational Journal of


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Quantum Chemistry


Organic Chemistry International

ElectrochemistryInternational Journal of



CatalystsJournal of


NH OO

NN

R

4a R =

4b NH2R =

OH4c R =

OH

4d R =

Cl4e R =

Cl

4f R =

4g CH3R =

4h NO2R =

4andashh

Figure 1 N-Phenyl anthranilic acid-based substituted 134-oxadiazole analogues (4andashh)

2 Material and Method

21 Preparation of Target Compounds Target compounds(4andashh) were prepared previously in our laboratory [7](Figure 1)The aromatic esterswere prepared by Fischer ester-ification Further these esters were subjected to formationof aryl hydrazide in presence of hydrazine hydrate Arylhydrazide 2a (1M)was dissolved in phosphorous oxychloride(5mL) and to it compound 3 (equimolar 1M) was addedThe reaction mixture after refluxing for 6-7 h was cooledto room temperature and poured onto crushed ice On neu-tralization of the contents with sodium bicarbonate solution(20) a solid mass separated out It was filtered and washedwith water It was crystallized by using methanol to give4a Similarly compounds 4bndashh were prepared The physicalproperties and spectral characterization were already dis-cussed [7]

22 Chemicals and Instruments All chemicals used in theestimation were of analytical grade Carrageenan was pur-chased from Sigma Chemicals A digital plethysmometer(model pth-7070 srnopt 070509 Medicad system) was usedfor measuring paw volume (mL) of rats edema Molecu-lar docking studies were carried out on Molecular VirtualDocker 500

23 Animals Albino rats of either sex around 180ndash200 gof weight were selected for both anti-inflammatory andanalgesic activity respectively The animals were kept inpolypropylene cages (3 in each cage) at an ambient tem-perature of 20 plusmn 2∘C and 55ndash65 relative humidity A 12-12 hrs light and dark schedule was maintained in the animalhouse The rats had free access to water and food Theexperimental protocol was approved by the InstitutionalAnimals Ethical Committee (MMCPIEC1016) and animalcare was done as per the guidelines of the committee forthe purpose of control and supervision of experiments onanimals (CPCSEA) Government of India

24 Acute Toxicity Studies It has been fund from the lit-erature survey that 134-oxadiazole derivatives in a dose of1000mgkg produce mortality (LD

50) in rats Soon tenth of

the LD50 that is 100mgkg (ED

50) was selected as a dose for

anti-inflammatory and analgesic activity [8]

25 In Vivo Anti-Inflammatory Activity In carrageenanmodel albino rats of all groups were treated with subcuta-neous injection of 01mL of 1 wv solution of carrageenaninto the subplantar region of the right hind paw The pawwas marked with permanent marker at the planter regionwhere the paw volume was to be measured The diclofenacsodium (10mgkg) and test compounds (100mgkg) weresuspended in 03 sodium carboxy methyl celluloseThe testcompounds and vehicle (control) were administered po withthe help of gastric cannula half an hour after the injection ofcarrageenan in subplanter region of right paw Mean normalpaw volume was measured 30min prior to carrageenaninjection by using plethysmometer Mean increase in the pawvolume for control group (after carrageenan injection) andtest group was measured at 1 hr 2 hr and 3 hr [9] Percentinhibition of inflammation after teststandard was calculatedusing the formula

inhibition =119881119888minus 119881119905

119881119888

times 100 (1)

where 119881119905is the of paw volume (mL) of teststandard com-

pound at corresponding time and 119881119888is the paw volume (mL)

of control

26 Analgesic Activity Tail immersion method is based onthe observation that morphine-like drugs selectively prolongthe reaction time of the typical tail withdrawal reflex inmice Albino rats were divided in twenty-six groups eachcontaining six animals The tail of mice was immersed (1-2 cm) in warm water kept constant at 55∘CThe reaction timewas recorded by stopwatch (the reaction time is the timetaken by the rats to flick their tails) The latent period of thetail flick response will be determined before and 15 30 60and 120min after drug administration [10]

27 Statistical Analysis The results were expressed as mean plusmnSEM for six animals in each group for anti-inflammatory andanalgesic activity All the grouped data was statistically evalu-atedHypothesis testingmethod included one-way analysis of














3 Results




0

02

04

06

08

1

12

Redu

ctio

n in

paw

vol

ume (

mL)

Target compounds

Con


After 3hours

Dic

lofe

nac

sodi

um


0

5

10

15

20

25

Reac

tion

time (

s)

Target compounds

Con


Dic

lofe

nac

sodi

umAfter 120min



4e 4e 998400

4f 4f 998400

















4 Discussion




(SC-S58)


(SC-S58998400)





4f minus76499









5 Conclusion


Acknowledgment


References






















Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of














3 Results




0

02

04

06

08

1

12

Redu

ctio

n in

paw

vol

ume (

mL)

Target compounds

Con


After 3hours

Dic

lofe

nac

sodi

um


0

5

10

15

20

25

Reac

tion

time (

s)

Target compounds

Con


Dic

lofe

nac

sodi

umAfter 120min



4e 4e 998400

4f 4f 998400

















4 Discussion




(SC-S58)


(SC-S58998400)





4f minus76499









5 Conclusion


Acknowledgment


References






















Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


4e 4e 998400

4f 4f 998400

















4 Discussion




(SC-S58)


(SC-S58998400)





4f minus76499









5 Conclusion


Acknowledgment


References






















Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


(SC-S58)


(SC-S58998400)





4f minus76499









5 Conclusion


Acknowledgment


References






















Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


5 Conclusion


Acknowledgment


References






















Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of










Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of

Research Article Anti-Inflammatory, Analgesic Evaluation...

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