Neuropharmacological Profile of Novel Benzopyran-2-One Derivatives in Experimental Mice
Abstract
The present study was aimed to assess the neuropharmacological properties of the novel Benzopyran-2-one (coumarin) derivatives i.e., 7-(2-(m-nitro)-phenyl thiazolidinyl)-4-methyl benzopyran-2-one (comp-I), 7-(2-(o,p-dichloro)-phenyl thiazolidinyl)-4-methyl-benzopyran-2-one (comp-II) and 7-(2-(p-methyl)-phenyl thiazolidinyl)-4-methyl-benzopyran-2-one (comp-III) by using the various pharmacological activities i.e., anticonvulsant, anxiolytic, spontaneous motor activity, skeletal muscle relaxant and analgesic activities along with assay of serum gamma amino butyric acid (GABA) levels. Methods: Investigation of all neuropharmacological profiles were evaluated by performing the various pharmacological activities using the different screening models, i.e., anticonvulsant activity by pentylenetetrazole (PTZ) induced seizure model, anxiolytic activity (exploratory behaviour) by evasion test, assay of GABA levels after anticonvulsant model was performed by using chromatographic technique, spontaneous motor activity (SMA) by actophotometer test, skeletal muscle relaxation by rotarod test and analgesic activity by Eddy’s hot plate and Haffner’s tail clip methods in Wistar albino mice. Results: All the Benzopyran-2-one derivatives showed significant anticonvulsant activity by prolonging the onset of seizures and reducing the duration of seizures and mortality rate when compared with control group. The test compounds showed significant anxiolytic effect. The serum GABA levels were significantly elevated in the treated groups, when compared with that of control group. The test compounds showed significant decrease in the spontaneous motor activity. The test compounds significantly showed a skeletal muscle relaxation after 30 min of dosing. The test compounds showed significant increase in paw licking time to heat stimuli in hot plate method and increase in the reaction time of mice to dislodge the clip. Conclusion: The present study has revealed that all the Benzopyran-2-one derivatives possess significant neuropharmacological properties confirmed by anticonvulsant, anxiolytic, skeletal muscle relaxant, analgesic effects and decrease in spontaneous motor activity at a dose of 20 mg/kg p.o. in swiss albino mice.
Full Text:
PDFReferences
Miyashiro KY, Bell TJ, Sul JY, Eberwine J (2009), “Subcellular Neuropharmacology: The importance of intracellular targeting”, Trends in Pharmacological Sciences, Volume 30, Issue 4, pp. 203−211.
(2003), “Opinion and evidence in Neurology and Psychiatry”, CNS Drugs, Volume 17, Issue 10, pp. 763−769.
Dinunzio JC, Williams RO (2008), “CNS Disorders- Current treatment options and the prospects for advanced therapies”, Drug Development and Industrial Pharmacy, Volume 34, pp. 1141−1167.
Brunton LL, Lazo JS, Parker KL (2006), “Goodman & Gilman's the pharmacologic basis of therapeutics”, 11th ed. New York: McGraw-Hill, pp. 429.
Conn PJ, Lindsley CW Jones CK (2008), “Activation of metabotropic glutamate receptors as a novel approach for the treatment of schizophrenia”, Trends in Pharmacological Sciences, Volume 30, Issue 1, pp. 25−31.
Rosenberg RN (2005), “Translational research on the way to effective therapy for Alzheimer disease”, Arch Gen Psychiatry, Volume 62, pp. 1186−1192.
Steiner M (2008), “Advances in neurobiology, assessment and treatment of female-specific mood disorders”, J Psychiatry Neurosci, Volume 33, Issue 4, pp. 289−290.
Cramer JA, leppik IE, Rue KD, Edrich P, Kramer G (2003), “Tolerability of levetiracetam in elderly patients with CNS disorders”, Epilepsy Research, Volume 56, pp. 135−145.
Abid M, Hrishikeshavan HJ, Asad M (2006), “Pharmacological evaluation of Pachyrrhizus erosus (L) seeds for central nervous system depressant activity”, Ind J Physiol Pharmacol, Volume 50, Issue 2, pp. 143−151.
Cooper J, Bloom F, Roth RH (2002), “The Biochemical Basis of Neuropharmacology”, Oxford University Press, pp. 48.
Meyer JS, Quenzer LS (2004), “Psychopharmacology; Drugs, the Brain and Behavior”, Sinauer Associates, ISBN: 0-87-893534-7.
Gomes PB, Noronha EC, De Melo CTV, Bezerra JNS, et al. (2008), “Central effects of isolated fractions from the root of Petiveria alliacea L. (tipi) in mice”, Journal of Ethnopharmacology, Volume 120, pp. 209−214.
Leal LKAM, Ferreira AAG, Bezerra GA, Matos FJA, Viana GSB (2000), “Antinociceptive, anti-inflammatory and bronchodilator activities of Brazilian medicinal plants containing coumarin a comparative study,. J Ethnopharmacol, Volume 70, pp. 151−159.
Al-Yousuf MH, Ali BH, Bashir AK,Tanira MOM, Blunden G (2002). “CNS activity of Leucas inflata in mice. Phytomedicine”, Volume 9, Issue 6, pp. 501−507.
Kang SY, Kim YC (2007), “Neuroprotective Coumarins from the root of Angelica gigas: Structure- activity relationships”, Arch Pharm Res; Volume 30, Issue 11, pp. 1368−1373.
Wakeel OK, Aziba PI, Ashorobi RB, Umukoro S, Aderibigbe AO, Awe EO (2004), “Neuropharmacological activities of Ficus platyphylla stem bark in mice”, African J Biomed Research, Volume 7, pp. 75−78.
Kumar RS, Sundram RS, Sivakumar P, Nethaji R, Senthil V, Murthy NV, et al. (2008), “CNS activity of the methanol extracts of Careya arborea in experimental animal model”, Bangladesh J Pharmacol, Volume 3, pp. 36−43.
Yadav AV, Kawale LA, Nade VS. (2008), “Effect of Morus alba L (mulberry) leaves on anxiety in mice”, Ind J Pharmacol, Volume 40, Issue 10, pp. 32−36.
Epifano F, Molinaro G, Genovese S, Ngomba RT, Nicoletti F, Massimo C. (2008), “Neuroprotective effect of prenyloxycoumarins from edible vegetables”, Neuroscience letters, Volume 443, pp. 57−60.
Kusanur RA, Ghate M, Kulkarni MV. (2004), “Synthesis of spiro [indolo-1, 5-benzodiazepines] from 3-acetyl coumarins for use as possible antianxiety agents”, J Chem Sci, Volume 116, Issue 5, pp. 265−270.
Bhat MA, Siddiqui N, Khan SA (2006), “Synthesis of novel thioureido derivatives of sulfonamides and Thiosemicarbazido derivatives of coumarin as potential anticonvulsant and analgesic agents”, Indian J Pharma Sci, Volume 68, Issue 1, pp. 120−124.
Ghate M, Kusanur RA, Kulkarni MV (2005), “Synthesis and in vivo analgesic and anti-inflammatory activities of some bi-heterocyclic coumarin derivatives”, Eur J Med Chem, Volume 40, pp. 882−887.
Amin KM, Rahman DEA, Al-Eryani YA (2008), “Synthesis and preliminary evaluation of some substituted coumarins as anticonvulsant agents”, Bioorg Med Chem, Volume 16, pp. 5377−5388.
Smyth T, Ramachandaran VN, Smyth WF (2009), “A study of the antimicrobial activity of selected naturally occuring and synthetic coumarins”, International journal of antimicrobial agents, Volume 30, pp. 421−426.
Bruneton J (1999), “Pharmacognosy and Phytochemistry of Medicinal Plants”, 2nd ed. Lavoisier publication, pp. 263−277.
Egan DA, Kennedy R, Moran E, Thornes RD (1990), “The pharmacology, metabolism, analysis and applications of coumarin and coumarin related compounds”, Drug Metab Rev, Volume 22, pp. 503–529.
Coumarin. IPCS 2002.
Ojala T. Remes S, Haansuu P, Vuorela H, et al. (2000), “Antimicrobial activity of some coumarin containing herbal plants growing in finland”, Journal of ethnopharmacology, Volume 73, pp. 299−305.
Guilet D, Seraphin D, Rondeau D, Richomme P, Bruneton J (2001), “Cytotoxic coumarins from calophyllum dispar”, Phytochemistry, Volume 58, pp. 571−575.
Ahmed AA, Hegazy MF, Hassan NM, Wojcinska M, Karchesy J, Pare PW (2006), “Constituents of Chrysothamnus viscidiflorus”, Phytochemistry, Volume 67, pp. 1547−1553.
Tada Y, Shikishima Y, Takaishi Y, et al. (2002), “Coumarins and γ-pyrone derivatives from prangos pabularia: antibacterial activity and inhibition of cytokine release”, Phytochemistry, Volume 59, pp. 649−654.
Bighetti AE, Antonio MA, Kohn LK, Rehder VLG et al. (2005), “Antiulcerogenic activity of a crude hydroalcoholic extract and coumarin isolated from Mikania Laevigata Schultz Bip. Phytomedicine”, Volume 12, pp. 72−77.
Ito C, Itoigawa M, Onoda S, Hosokawa A, et al. (2005) “Chemical constituents of Murraya Siamensis: three coumarins and their antitumor promoting effect”, Phytochemistry, Volume 66, pp. 567−572.
Stein AC, Alvarez S, Avancini C, Zacchino S, Poser GV (2006), “Antifungal activity of some coumarins obtained from species of Pterocaulon (Asteraceae)”, Journal of ethnopharmacology, Volume 107, pp. 95−98.
Leu CH, Li CY, Yao X, Wu TS (2006), “Constituents from the leaves of Phellodendron amurense and their antioxidant activity”, Chem. pharm. Bull, Volume 54, Issue 9, pp. 1308−1311.
Wu CR, Huang MY, Lin YT, Ju HY, Ching H (2007), “Antioxidant properties of Cortex Frexini and its simple coumarins”, Food chemistry, Volume 104, pp. 1464−1471.
Kim YA, Kong CS, Yea SS, Seo Y (2009), “Constituents of corydalis heterocarpa & their antiproliferative effect on human cancer cells.food and chemical toxicology”, (article in press).
Hu XJ, Jin HZ, Xu WZ, Chen M, et al. (2008), “Anti-inflammatory and analgesic activities of Edgeworthia chrysantha and its effective chemical constituents”, Boil. Pharm. Bull. Volume 31, Issue 9, pp. 1761−1765.
Kang KH, Kong CS, Seo Y, Kim MM, Kim SK (2009), “Antiinflammatory effect of coumarins isolated from corydolis heterocarpa in HT-29 human colon carcinoma cells”, Food and chemical toxicology, Volume 47, pp. 2129−2134.
Khan AU, Gilani AH (2009), “Antispasmodic and bronchodilator activities of Artemisia Vulgaris are mediated through dual blockade of muscarinic receptors and calcium influx, “Journal of ethnopharmacology”, Volume 126, pp. 480−486.
Ziyyat A, Legssyer A, Mekhfi H, Dassouli A, Serhrouchni M, Benjelloun W (1997), “Phytotherapy of hypertension and diabetes in oriental Morocco”, J Ethnopharmacol, Volume 58, Issue 1, pp. 45−54.
Pari L, Rajarajeswari N (2009), “Efficacy of coumarin on hepatic key enzymes of glucose metabolism in chemical induced type 2 diabetic rats”, Chemico-Biological Interactions, Volume 181, pp. 292–296.
Dongmoa AB, Azebaze AGB, Nguelefack TB, Ouahouod BM, Sontia B, Meyerf M, Nkengfack AE et al. (2007), “Vasodilator effect of the extracts and some coumarins from the stem bark of Mammea africana (Guttiferae)”, J Ethnopharmacol, Volume 111, pp. 329–334.
Hansel R, Keller K, Rimpler H, Schneider G. (eds.; 1993), Hagers Handbuch der pharmazeutischen Praxis, Drogen E-O, Band 5, 5. Aufl. p. 664-670. Springer-Verlag, Berlin, Germany.
Lewis HM (1994), “Therapeutic progress II: treatment of psoriasis”, J Clin Pharm Therap, Volume 19, pp. 223−232.
Iqbal PF, Bhat AR, Azam A (2009), “Antiamoebic coumarins from the root bark of Adina cordifolia and their new thiosemicarbazone derivatives”, Eur J Med Chem, Volume 44, pp. 2252−2259.
Yoshikawa M, Nishida N, Ninomiya K, Ohgushi T, Kubo M, Morikawa T et al. (2006), “Inhibitory effects of coumarin and acetylene constituents from the roots of Angelica furcijuga on D galactosamine/lipopolysaccharide induced liver injury in mice and on nitric oxide production in lipopolysaccharide-activated mouse peritoneal macrophages”, Bioorg Med Chem, Volume 14, pp. 456−463.
Auzi ARA, Hawisa NT, Sherif FM, Sarker SD (2007), “Neuropharmacological properties of Launaea resedifolia”, Brazilian J Pharmacog, Volume 17, pp. 160−165.
(2007), “Coumarin derivatives from Loeselia Mexicana. Determination of anxiolytic activity of Daphnoretin on elevated plus maze”, J Mex Chem Soc, Volume 51, Issue 4, pp. 193−197.
Yang YZ, Ranz A, Pan HZ, Zhang ZN, Lin XB, Meshnick SR. (1992), “Daphnetin: a novel antimalarial agent with in vitro and in vivo activity”, Am J Trop Med Hyg, Volume 46, Issue 1, pp. 15−20.
Oliver GF, Winkelmann RK (1993), “Treatment of lichen planus. Drugs”, Volume 45, Issue 1, pp. 56−65.
Prasad RY, Kumar RP, Deepti ACH, Ramana MV (2006), “Synthesis and antimicrobial activity of some novel chalcones of 2-Hydroxy-1- acetonapthone and 3-Acetyl Coumarin”, Eur J Med Chem, Volume 3, Issue 13, pp. 236−241.
Reddy NS, Mallireddigari MR, Cosenza S, Gumireddy K, Bell SC, Reddy EP, et al. (2004), “Synthesis of new coumarin 3-(N-aryl) sulfonamides and their anticancer activity”, Bioorg Med Chem, Volume 14, pp. 4093−4097.
Karali N, Kocabalkanli A, Gursoy A, Ates O (2002), “Synthesis and antitubercular activity of 4 - (3 - coumarinyl) – 3 – cyclohexyl – 4 – thiazolin – 2 - one benzylidenehydrazones. IL Farmaco”, Volume 57, pp. 589−593.
Maddi V, Raghu KS, Rao MNA (1992), “Synthesis and anti-inflammatory activity of 3-(Benzylideneamino) coumarin in rodents”, J Pharm Sci, Volume 81, pp. 964−966.
Su CX, Mouscadet JF, Chang CC, Tsai HJ, Hsu LY. (2006), “HIV-1 Integrase Inhibition of Biscoumarin Analogues”, Chem Pharm Bull, Volume 54, Issue 5, pp. 682−686.
Alvey L, Prado S, Huteau V, Saint-Joanis B, Michel S, Koch M, et al. (2008), “A new synthetic access to furo[3,2-f]chromene analogues of an antimycobacterial”, Bioorg Med Chem, Volume 16, pp. 8264−8272.
Kalkhambkar RG, Kulkarni GM, Kamanavalli CM, Premkumar N, Asdaq SMB, Sun CM (2008), “Synthesis and biological activities of some new fluorinated coumarins and 1-aza coumarins”, Eur J Med Chem, Volume 43, pp. 2178−2188.
Tripathi RP, Tiwari VK, Misra-Bhattacharya S, Tyagi K, Srivastava, Murty PK (2003), “7-O-[4-methyl piperazine-1-(2-acetyl)]-2H-1-benzopyran-2-one: a novel antifilarial lead compound”, Acta Tropica, Volume 87, pp. 215−224.
Manolov I, Maichle-Moessmer C, Danchev N (2006), “Synthesis, structure, toxicological and pharmacological investigations of 4-hydroxycoumarin derivatives”, Eur J Med Chem, Volume 41, pp. 882−890.
Chimenti F, Bizzarri B, Bolasco A, Secci D, Chimenti P, Carradori.S Synthesis and in vitro selective anti-Helicobacter pylori activity of N-substituted-2-oxo-2H-1-benzopyran-3-carboxamides. Eur J Med Chem 2006;4:208-12.
Sardari S, Mori Y, Horita K, Micetich RG, Nishibe S, Daneshtalab M (1999), “Synthesis and antifungal activity of coumarins and angular furanocoumarins”, Bioorganic and Medicinal Chemistry, Volume 7, pp. 1933−1940.
Kim SN, Kim NH, Park YS, Kim H, Lee S, Wang Q, Kim YK (2009), “7-Diethylamino-3(2’-benzoxazolyl)-coumarin is a novel microtubule inhibitor with antimitotic activity in multidrug resistant cancer cell”, Biochemical pharmacology, Volume 77, pp. 1773−1779.
Symeonidis T, Fylaktakidou KC, Litina DJH, Litinas KE (2009), “Synthesis and anti-inflammatory evaluation of novel angularly or linearly fused coumarins”, European journal of medicinal chemistry, Volume 44, pp. 5012−5017.
Creaven BS, Devereux M, Karcz D, Kellett A, et al. (2009), “Copper (II) complexes of coumarin derived Schiff bases and their anticandida activity”, Journal of inorganic biochemistry, Volume 103, pp. 1196−1203.
Matos MJ, Vina D, Picciau C, Orallo F, Santana L, Uriarte E (2009), “Synthesis and evaluation of 6-methyl-3-phenylcoumarins as potent and selective MAO-B inhibitors”, Bioorganic and medicinal chemistry letters, Volume 19, pp. 5053−5055.
Pierson JT, Dumetre A, Hutter S, Delmas F, Laget M, et al. (2009), “Synthesis and antiprotozoal activity of 4-arylcoumarins”, European journal of medicinal chemistry, (Article in press).
Cavar S, Kovac F, Maksimovic M. (2009), “Synthesis and antioxidant activity of selected 4- methyl coumarins”, Food chemistry, Volume 117, pp. 135−142.
Tyagi YK, Kumar A, Raj HG, Vohra P, Gupta G, Kumari R, et al. (2005), “Synthesis of novel amino and acetyl amino-4-methylcoumarins and evaluation of their antioxidant activity”, Eur J Med Chem, Volume 40, pp. 413−420.
Laurin P, Ferroud D, Klich M, Dupuis-Hamelin C, Mauvais P, Lassaigne P, et al. (1999), “Synthesis and in vitro evaluation of novel highly potent coumarin inhibitors of gyrase B”, Bioorg Med Chem Lett, Volume 9, pp. 2079−2084.
Grimm EL, Brideau C, Chauret N. (2006), “Substituted coumarins as potent 5-lipoxygenase inhibitors”, Bioorg Med Chem Lett, Volume 16, pp. 2528−2531.
Radanyi C, Bras GL, Messaoudi S, Bouclier C, Peyrat JF, Brion JD, et al. (2008), “Synthesis and biological activity of simplified denoviose-coumarins related to novobiocin as potent inhibitors of heat-shock protein 90 (hsp90)”, Bioorg Med Chem Lett, Volume 18, pp. 2495−2498.
Raad I, Terreux R, Richomme P, Matera EL, Dumontet C, Raynaud J, et al. (2006), “Structure–activity relationship of natural and synthetic coumarins inhibiting the multidrug transporter P-glycoprotein”, Bioorg Med Chem, Volume 14, pp. 6979−6987.
Han S, Zhou V, Pan S, Liu Y, Hornsby M, McMullan D, et al (2005), “Identification of coumarin derivatives as a novel class of allosteric MEK1 inhibitors”, Bioorg Med Chem Lett, Volume 15, pp. 5467−5473.
Ferrari AM, Sgobba M, Gamberini MC, Rastelli G (2007), “Relationship between quantum-chemical descriptors of proton dissociation and experimental acidity constants of various hydroxylated coumarins. Identification of the biologically active species for xanthine oxidase inhibition”, Eur J Med Chem, Volume 42, pp. 1028−1031.
Lin SL, Kuo PY, Yang DY (2007), “Design and Synthesis of a Coumarin-based Acidichromic Colorant”, Molecules, Volume 12, pp. 1316−1324.
Ronad PM, Noolvi MN, Sapkal S, Dharbhamulla S, Maddi VS (2010), “Synthesis and antimicrobial activity of 7-(2-substituted phenylthiazolidinyl)-benzopyran-2-onederivatives”, Eur J Med Chem, Volume 45, Issue 1, pp. 85−89.
Organization for Economic Co-operation and Development, revised draft Guidelines 423. OECD guideline for the testing of Chemicals. Revised Document. October 2000.
Olayiwola G, Obafemi CA, Taiwo FO (2007), “Synthesis and neuropharmacological activity of some quinoxalinone derivatives”, Afr J Biotech, Volume 6, Issue 6, pp. 777−786.
Adzu B, Amos S, Muazzam I, Inyang US, Gamaniel KS (2002), “Neuropharmacological screening of Diospyros mespiliformis in mice”, J Ethnopharmacol, Volume 83, pp. 139−143.
Nassiri-Asl, Zamansoltani F, Torabinejad B (2009), “Antiepileptic effects of quinine in pentylenetetrazole model of seizure”, British Epilepsy Association (Seizure), Volume 18, Issue 2, pp. 129−132.
Viswanatha Swamy AHM, Thippeswamy AHM, Manjula DV, Mahendra Kumar CB (2006), “Some Neuropharmacological Effects of the Methanolic Root Extract of Cissus quadrangularis in mice”, Afr J Biomed Res, Issue 9, pp. 69−76.
Turner RA. (1965), “Screening methods in pharmacology”, Academic Press, New York.
Mishraa OP, Singhala D, Upadhyayb RS, Prasad R, Atri D. (2007), “Cerebrospinal fluid zinc, magnesium, copper and gamma-aminobutyric acid levels in febrile seizures, “J Pediatric Neurology”, Volume 5, pp. 39−44.
Shalam Md, Shantakumar SM, Narasu ML (2007), “Neuropharmacological Profile of Trans-01 a Polyherbal Formulation in Mice”, Pharmacol online, Volume 1, pp. 146−151.
S.K. Kulkarni (1999), “In: Handbook of Experimental Pharmacology”, 3rd ed. New Delhi: Vallabh prakashan Publishers;.pp. 117−122.
Kuribara H, Higuchi Y, Tadokoro S (1977), “Effects of central depressants on rota-rodand traction performances in mice”, Japan J Pharmacol, Volume 27, pp. 117−126.
Eddy NB, Leimbach DJ (1953), “J Pharmacol Exptl Therap”, Volume 107, pp. 385.
Malairajan P, Gopalakrishnan G, Narasimhan S, Veni KJK (2006), “Analgesic activity of some Indian medicinal plants”, J Ethnopharmacol, Volume 106, pp. 425−428.
Mate GS, Naikwade NS, Magdum CS, Chowki AA, Patil SB (2008), “Evaluation of anti-nociceptive activity of Cissus quadrangularis on albino mice”, Int J Green Pharm, pp. 118−121.
Palanichamy S, Nagarajan S (1990), “Analgesic activity of cassia alata leaf extract and kaempferol 3-o-sophoroside”, J Ethnopharmacol, Volume 29, Issue 1, pp. 73−78.
Ojewole JAO (2008), “Anticonvulsant effect of Rhus chirindensis (Baker F)
(Anacardiaceae) stem-bark aqueous extract in mice”, J Ethnopharmacol, Volume 117, pp. 130−135.
Sayyah M, Mandgary A, Kamalinejad M (2002), “Evaluation of the anticonvulsant activity of the seed acetone extract of Ferula gummosa Boiss against seizures induced by pentylenetetrazole and electroconvulsive shock in mice”, J Ethnopharmacol, Volume 82, pp. 105−109.
Sarker SD, Uddin SJ, Shilpi JA, Rouf R, Ferdous MM, Nahar L. (2007), “Neuropharmacological properties of Xylocarpus moluccensis”, Fitoterapia, Volume 78, pp. 107−111.
Yemitan OK, Salahdeen HM. (2005), “Neurosedative and muscle relaxant activities of aqueous extract of Bryophyllum pinnatum”, Fitoterapia, Volume 76, pp. 187−193.
Yadav AV, Kawale LA, Nade VS (2008), “Effect of Morus alba L (mulberry) leaves on anxiety in mice”, Ind J Pharmacol, Volume 40, Issue 10, pp. 32−36.
Shalam Md, Shantakumar SM, Narasu ML. (2007), “Neuropharmacological Profile of Trans-01 a Polyherbal Formulation in Mice”, Pharmacol online, Volume 1, pp. 146−151.
Perez RMG, Perez JAL, Garcia LMD, Sossa HM (1998), “Neuropharmacological activity of Solanum nigrum fruit”, J Ethnopharmacol, Volume 62, pp. 43−48.
Jaishree V, Badami S, Kumar MR, Tamizhani T. (2008), “Antinociceptive activity of swertiamarin isolated from Enicostemma axillare”, Phytomedicine, (Article in press).
Bianchi C, Franceschini J. (1954), “Experimental observations on Haffner’s method for testing analgesic drugs”, Brit J Pharmacol, Volume 9, pp. 280−284.
Adeyemi OO, Okpo SO, Okpaka O (2004), “The analgesic effect of the methanolic extract of Acanthus montanus”, J Ethnopharmacol, Volume 90, pp. 45−48.
Refbacks
- There are currently no refbacks.