{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T02:26:11Z","timestamp":1760235971781,"version":"build-2065373602"},"reference-count":54,"publisher":"MDPI AG","issue":"20","license":[{"start":{"date-parts":[[2021,10,13]],"date-time":"2021-10-13T00:00:00Z","timestamp":1634083200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001655","name":"Deutscher Akademischer Austauschdienst","doi-asserted-by":"publisher","award":["57369155"],"award-info":[{"award-number":["57369155"]}],"id":[{"id":"10.13039\/501100001655","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002347","name":"Bundesministerium f\u00fcr Bildung und Forschung","doi-asserted-by":"publisher","award":["01DG17008B"],"award-info":[{"award-number":["01DG17008B"]}],"id":[{"id":"10.13039\/501100002347","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>The leaves of Ranunculus multifidus Forsk. are traditionally used for the treatment of malaria in several African countries. In the present study, 80% methanol (RM-M) and hydrodistilled (RM-H) extracts of fresh leaves from R. multifidus and its major constituent anemonin were tested for their in vivo antimalarial activity against Plasmodium berghei in mice. Anemonin was also tested for its in vitro antimycobacterial activity against Mycobacterium smegmatis and M. abscessus in a microbroth dilution assay, and bacterial growth was analyzed by OD measurement. The isolation of anemonin from RM-H was carried out using preparative thin layer chromatography (PTLC). The chemical structures of anemonin and its hydrolysis product were elucidated using spectroscopic methods (HR\u2013MS; 1D and 2D-NMR). Results of the study revealed that both RM-M and RM-H were active against P. berghei in mice, although the latter demonstrated superior activity (p &lt; 0.001), as compared to the former. At a dose of 35.00 mg\/kg\/day, RM-H demonstrated a chemosuppression value of 70% in a 4-day suppressive test. In a 4-day suppressive, Rane\u2019s and prophylactic antimalarial tests, anemonin showed median effective doses (ED50s) of 2.17, 2.78 and 2.70 \u03bcM, respectively. However, anemonin did not inhibit the growth of M. smegmatis and M. abscessus.<\/jats:p>","DOI":"10.3390\/molecules26206179","type":"journal-article","created":{"date-parts":[[2021,10,13]],"date-time":"2021-10-13T21:48:39Z","timestamp":1634161719000},"page":"6179","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["In Vivo Antimalarial Activity of Leaf Extracts and a Major Compound Isolated from Ranunculus multifidus Forsk"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9469-7192","authenticated-orcid":false,"given":"Betelhem","family":"Sirak","sequence":"first","affiliation":[{"name":"Department of Pharmacy, College of Medicine and Health Sciences, Arba Minch University, Arba Minch P.O. Box 21, Ethiopia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lea","family":"Mann","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Chemistry, Faculty of Natural Sciences, Martin-Luther-Universitaet Halle-Wittenberg, 3, 06120 Halle (Saale), Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Adrian","family":"Richter","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Chemistry, Faculty of Natural Sciences, Martin-Luther-Universitaet Halle-Wittenberg, 3, 06120 Halle (Saale), Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kaleab","family":"Asres","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Chemistry and Pharmacognosy, School of Pharmacy, College of Health Sciences, Addis Ababa University, Addis Ababa P.O. Box 1176, Ethiopia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Peter","family":"Imming","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Chemistry, Faculty of Natural Sciences, Martin-Luther-Universitaet Halle-Wittenberg, 3, 06120 Halle (Saale), Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,10,13]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"World Health Organization (2020). World malaria report 2020: 20 years of global progress and challenges. World Malaria Report 2020: 20 Years of Global Progress and Challenges, World Health Organization.","DOI":"10.30875\/60123dd4-en"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"405","DOI":"10.1186\/1475-2875-13-405","article-title":"Clinical malaria case definition and malaria attributable fraction in the highlands of western Kenya","volume":"13","author":"Afrane","year":"2014","journal-title":"Malar. J."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1016\/S1473-3099(04)01043-6","article-title":"The global distribution and population at risk of malaria: Past, present, and future","volume":"4","author":"Hay","year":"2004","journal-title":"Lancet Infect."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1186\/s40794-019-0090-z","article-title":"Burden of malaria in Ethiopia, 2000\u20132016: Findings from the Global Health Estimates 2016","volume":"5","author":"Girum","year":"2019","journal-title":"Trop. Dis. Travel Med. Vaccines"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"8863002","DOI":"10.1155\/2021\/8863002","article-title":"Prevalence of malaria among adults in Ethiopia: A systematic review and meta-analysis","volume":"2021","author":"Kendie","year":"2021","journal-title":"J. Trop. Med."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1","DOI":"10.9734\/EJMP\/2016\/25670","article-title":"An overview of Ethiopian traditional medicinal plants used for cancer treatment","volume":"14","author":"Abebe","year":"2016","journal-title":"Eur. J. Med. Plants"},{"key":"ref_7","first-page":"1","article-title":"Ethnopharmacological survey of medicinal plants used against malaria in Butembo City (DR Congo)","volume":"1","author":"Kasali","year":"2014","journal-title":"J. Adv. Bot. Zool."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2","DOI":"10.4172\/2573-4555.1000271","article-title":"Ethnobotany of medicinal plants used to treat leishmaniasis in Ethiopia: A systematic review","volume":"7","author":"Aschale","year":"2018","journal-title":"J. Tradit. Med. Clin. Naturopat."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1","DOI":"10.36876\/smjmps.1006","article-title":"Ethnobotanical study of medicinal plants used to treat human diseases in Enarj Enawga district, East Gojjam zone, Amhara region, Ethiopia","volume":"1","author":"Birhan","year":"2017","journal-title":"SM J. Med. Plant Stud."},{"key":"ref_10","first-page":"258","article-title":"Ethnobotanical study on traditional medicinal plants in Dega Damot woreda, Amhara Region, North Ethiopia","volume":"5","author":"Limenih","year":"2015","journal-title":"Int. J. Res. Pharm. Chem."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1016\/j.sajb.2014.06.009","article-title":"Traditional herbal remedies used by women in a rural community in northern Maputaland (South Africa) for the treatment of gynecology and obstetrics complaints","volume":"94","author":"Ngubane","year":"2014","journal-title":"S. Afr. J. Bot."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"656","DOI":"10.1016\/j.jep.2013.07.018","article-title":"Plants traditionally used individually and in combination to treat sexually transmitted infections in northern Maputaland, South Africa: Antimicrobial activity and cytotoxicity","volume":"149","author":"Naidoo","year":"2013","journal-title":"J. Ethnopharmacol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1099","DOI":"10.1016\/0031-9422(93)85030-U","article-title":"Protoanemonin in Australian Clematis","volume":"33","author":"Southwell","year":"1993","journal-title":"Phytochemistry"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1016\/j.jep.2004.02.011","article-title":"In vitro antiplasmodial activity of medicinal plants native to or naturalised in South Africa","volume":"92","author":"Clarkson","year":"2004","journal-title":"J. Ethnopharmacol."},{"key":"ref_15","first-page":"62","article-title":"Plasmodium berghei ANKA infection in ICR mice as a model of cerebral malaria","volume":"7","author":"Basir","year":"2012","journal-title":"Iran. J. Parasitol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"261","DOI":"10.2174\/138945009787581203","article-title":"Testing of natural products and synthetic molecules aiming at new antimalarials","volume":"10","author":"Krettli","year":"2009","journal-title":"Curr. Drug Targets"},{"key":"ref_17","first-page":"325","article-title":"Isolation of anemonin from Pulsatilla wallichiana and its biological activities","volume":"41","author":"Ali","year":"2019","journal-title":"J. Chem. Soc. Pak."},{"key":"ref_18","first-page":"1","article-title":"Synthesis and constitution of anemonin","volume":"455","author":"Asahina","year":"1920","journal-title":"J. Pharm. Soc. Jpn."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1007\/s12272-015-0691-7","article-title":"Antioxidant C-glycosylflavones of Drymaria cordata (Linn.) Willd","volume":"39","author":"Nono","year":"2016","journal-title":"Arch. Pharmacal Res."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1483","DOI":"10.5650\/jos.ess18056","article-title":"Tunisian Clematis flammula essential oil enhances wound healing: GC-MS analysis, biochemical and histological assessment","volume":"67","author":"Saidi","year":"2018","journal-title":"J. Oleo Sci."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1016\/j.jep.2009.01.010","article-title":"How scientific is the science in ethnopharmacology? Historical perspectives and epistemological problems","volume":"122","author":"Gertsch","year":"2009","journal-title":"J. Ethnopharmacol."},{"key":"ref_22","first-page":"551","article-title":"Glutathione\u2013functions and metabolism in the malarial parasite Plasmodium falciparum","volume":"384","author":"Becker","year":"2003","journal-title":"J. Biol. Chem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"569","DOI":"10.1039\/a605727f","article-title":"Significance of the intramolecular transformation of glutathione thiyl radicals to \u03b1-aminoalkyl radicals. Thermochemical and biological implications","volume":"2","author":"Zhao","year":"1997","journal-title":"J. Chem. Soc. Perkin Trans."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"855","DOI":"10.1016\/0006-2952(88)90172-4","article-title":"Glutathione reductase inhibitors as potential antimalarial drugs: Effects of nitrosoureas on Plasmodium falciparum in vitro","volume":"37","author":"Zhang","year":"1988","journal-title":"Biochem. Pharmacol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1006\/abio.1998.3061","article-title":"Synthesis of 5,5\u2032-dithiobis (2-nitrobenzamides) as alternative substrates for trypanothione reductase and thioredoxin reductase: A microtiter colorimetric assay for inhibitor screening","volume":"268","author":"Becker","year":"1999","journal-title":"Anal. Biochem."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1016\/S0014-5793(98)00031-3","article-title":"Recombinant Plasmodium falciparum glutathione reductase is inhibited by the antimalarial dye methylene blue","volume":"422","author":"Arscott","year":"1998","journal-title":"FEBS Lett."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"2671","DOI":"10.1128\/AAC.39.12.2671","article-title":"Antimalarial dyes revisited: Xanthenes, azines, oxazines, and thiazines","volume":"39","author":"Vennerstrom","year":"1995","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"711","DOI":"10.1146\/annurev.bi.52.070183.003431","article-title":"Glutathione","volume":"52","author":"Meister","year":"1983","journal-title":"Annu. Rev. Biochem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1016\/S0753-3322(03)00043-X","article-title":"The importance of glutathione in human disease","volume":"57","author":"Townsend","year":"2003","journal-title":"Biomed. Pharmacother."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2135","DOI":"10.1093\/jn\/127.11.2135","article-title":"Methionine and cysteine affect glutathione level, glutathione-related enzyme activities and the expression of glutathione S-transferase isozymes in rat hepatocytes","volume":"127","author":"Wang","year":"1997","journal-title":"J. Nutr."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1007\/BF00437648","article-title":"Antimicrobial activity of protoanemonin, a lactone from ranunculaceous plants","volume":"98","author":"Mares","year":"1987","journal-title":"Mycopathologia"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"333","DOI":"10.1021\/jm00213a003","article-title":"Antitumor agents. 21. A proposed mechanism for inhibition of cancer growth by tenulin and helenalin and related cyclopentenones","volume":"20","author":"Hall","year":"1977","journal-title":"J. Med. Chem."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1991","DOI":"10.1021\/ja01227a037","article-title":"The mechanism of action of antibiotics. The reaction of unsaturated lactones with cysteine and related compounds","volume":"67","author":"Cavallito","year":"1945","journal-title":"J. Am. Chem. Soc."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1111\/j.1574-6968.2008.01467.x","article-title":"Characterization of Trypanosoma cruzi L-cysteine transport mechanisms and their adaptive regulation","volume":"292","author":"Canepa","year":"2009","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"e00828-18","DOI":"10.1128\/AAC.00828-18","article-title":"Screening of preselected libraries targeting Mycobacterium abscessus for drug discovery","volume":"62","author":"Richter","year":"2018","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_36","unstructured":"Institute for Laboratory Animal Research (ILAR) (1996). Guide for the Care and Use of Laboratory Animals, The National Academy Press."},{"key":"ref_37","unstructured":"Organization for Economic Co-operation and Development (OECD) (2008). Guideline for the Testing of Chemicals: Acute Oral Toxicity-Up-and-Down Procedure, Organization for Economic Co-operation and Development (OECD)."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1260","DOI":"10.1016\/j.bmcl.2006.12.007","article-title":"Weak base dispiro-1,2,4-trioxolanes: Potent antimalarial ozonides","volume":"17","author":"Tang","year":"2007","journal-title":"Bioorg. Med. Chem. Lett."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"509","DOI":"10.1038\/nrd1416","article-title":"Antimalarial drug discovery: Efficacy models for compound screening","volume":"3","author":"Fidock","year":"2004","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"236","DOI":"10.1016\/j.jep.2005.08.006","article-title":"In vivo antimalarial activities of extracts from Amaranthus spinosus L. and Boerhaavia erecta L. in mice","volume":"103","author":"Hilou","year":"2006","journal-title":"J. Ethnopharmacol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"573","DOI":"10.1136\/pgmj.49.574.573","article-title":"Antimalarial drugs and their actions","volume":"49","author":"Peters","year":"1973","journal-title":"Postgrad. Med. J."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1016\/j.jep.2005.02.017","article-title":"The in vitro and in vivo antimalarial activity of Cardiospermum halicacabum L. and Momordica foetida Schumch. Et Thonn","volume":"99","author":"Waako","year":"2005","journal-title":"J. Ethnopharmacol."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Mergia, E., Shibeshi, W., Terefe, G., and Teklehaymanot, T. (2016). Antitrypanosomal activity of Verbascum sinaiticum Benth. (Scrophulariaceae) against Trypanosoma congolense isolates. BMC Complement. Altern. Med., 16.","DOI":"10.1186\/s12906-016-1346-z"},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Tamiru, W., Engidawork, E., and Asres, K. (2012). Evaluation of the effects of 80% methanolic leaf extract of Caylusea abyssinica (fresen.) fisch. & Mey. on glucose handling in normal, glucose loaded and diabetic rodents. BMC Complement. Altern. Med., 12.","DOI":"10.1186\/1472-6882-12-151"},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Misganaw, D., Engidawork, E., and Nedi, T. (2019). Evaluation of the antimalarial activity of crude extract and solvent fractions of the leaves of Olea europaea (Oleaceae) in mice. BMC Complement. Altern. Med., 19.","DOI":"10.1186\/s12906-019-2567-8"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"2598","DOI":"10.1128\/AAC.47.8.2598-2605.2003","article-title":"In vivo antimalarial activities of mono- and bis-quaternary ammonium salts interfering with Plasmodium phospholipid metabolism","volume":"47","author":"Ancelin","year":"2003","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Fentahun, S., Makonnen, E., Awas, T., and Giday, M. (2017). In vivo antimalarial activity of crude extracts and solvent fractions of leaves of Strychnos mitis in Plasmodium berghei infected mice. BMC Complement. Altern. Med., 17.","DOI":"10.1186\/s12906-016-1529-7"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1080\/00034983.1970.11686683","article-title":"The antimalarial activity of some quinolone esters","volume":"64","author":"Ryley","year":"1970","journal-title":"Ann. Trop. Med. Parasitol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"2515690X19890866","DOI":"10.1177\/2515690X19890866","article-title":"Evaluation of antimalarial activity of the 80% methanolic stem bark extract of Combretum molle against Plasmodium berghei in mice","volume":"24","author":"Mulaw","year":"2019","journal-title":"J. Evid.-Based Integr. Med."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"9217835","DOI":"10.1155\/2018\/9217835","article-title":"In vivo antimalarial activity of the 80% methanolic root bark extract and solvent fractions of Gardenia ternifolia Schumach. and Thonn. (Rubiaceae) against Plasmodium berghei","volume":"2018","author":"Nureye","year":"2018","journal-title":"Evid.-Based Complement. Altern. Med."},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Bantie, L., Assefa, S., Teklehaimanot, T., and Engidawork, E. (2014). In vivo antimalarial activity of the crude leaf extract and solvent fractions of Croton macrostachyus Hocsht. (Euphorbiaceae) against Plasmodium berghei in mice. BMC Complement. Altern. Med., 14.","DOI":"10.1186\/1472-6882-14-79"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"2515690X20984287","DOI":"10.1177\/2515690X20984287","article-title":"Antimalarial activity of seed extracts of Schinus molle against Plasmodium berghei in mice","volume":"26","author":"Mekuria","year":"2021","journal-title":"J. Evid.-Based Integr. Med."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1016\/0014-4894(65)90014-7","article-title":"Drug resistance in Plasmodium berghei Vincke and Lips, 1948. III. Multiple drug resistance","volume":"17","author":"Peters","year":"1965","journal-title":"Exp. Parasitol."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1080\/00034983.1992.11812694","article-title":"The chemotherapy of rodent malaria. Studies on pyronaridine and other Mannich base antimalarials","volume":"86","author":"Peters","year":"1992","journal-title":"Ann. Trop. Med. Parasitol."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/26\/20\/6179\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:12:42Z","timestamp":1760166762000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/26\/20\/6179"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,10,13]]},"references-count":54,"journal-issue":{"issue":"20","published-online":{"date-parts":[[2021,10]]}},"alternative-id":["molecules26206179"],"URL":"https:\/\/doi.org\/10.3390\/molecules26206179","relation":{},"ISSN":["1420-3049"],"issn-type":[{"type":"electronic","value":"1420-3049"}],"subject":[],"published":{"date-parts":[[2021,10,13]]}}}