Kutaja (Holarrhena Antidysentrica) – Part-1

Recent Research on Holarrhena antidysentrica

• The present study shows the evaluation of the anti-malarial effects of two plants commonly used against malaria in the Garhwal region of north-west Himalaya, to discover herbal-based medicine. The present investigation reflects the use of these traditional medicinal plants against malaria and these plants may work as potential sources in the development of a variety of herbal formulations for the treatment of malaria. Verma G, Dua VK, Agarwal DD, Atul PK. Anti-malaria activity of Holarrhena anti- dysenterica and Viola canescens, plants traditionally used against malaria in the Garhwal region of north-west Himalaya. Malar J. 2011 Feb 2; 10: 20. do: 10. 1186/ 1475- 2875- 10- 20.
• The alkaloids from the ethanolic extract of H. anti- dysenterica seeds were evaluated for their anti-bacterial activity against clinical isolates of enteropathogenic Escherichia coli (EPEC) in vitro, and their anti-diarrheal activity on castor oil-induced diarrhea in rats, in vivo. The results suggest the usefulness of alkaloids of H. anti- dysentrica seeds as antibacterial and anti- diarrhoeal agents. Kavitha D, Shilpa PN, Devaraj SN. Antibacterial and anti-diarrhoeal effects of alkaloids of Holarrhena anti- dysenterica WALL. Indian J Exp Biol. 2004 Jun; 42 (6): 589- 94.
• Sharma, Vikas & Hussain, Shabir & Bakshi, Manish & Bhat, Neha & Saxena, Ajit. (2014). In vitro cytotoxic activity of leaves extracts of Holarrhena antidysenterica against some human cancer cell lines. Indian Journal of Biochemistry & Biophysics. 51. 46-51. In vitro, the cytotoxic potential of extracts (95% and 50% ethanolic extract and hot water extract at a concentration of 100 microg/ml) from leaves of Holarrhena antidysenterica was evaluated against fourteen human cancer cell lines- A- 549, COLO- 205, DU- 145, HeLa, HEP- 2, IMR- 32, KB, MCF- 7, NCI- H23, OVCAR- 5, SiHa, SK- N- MC, SW- 620 and ZR- 75- 1 from nine different tissues (breast, colon, cervix, CNS, lung, liver, oral, ovary and prostate) using SRB assay. The 95% ethanolic extract displayed a maximum anti-proliferative effect in the range of 73- 92% against eight human cancer cell lines, while 50% ethanolic extract showed cytotoxic activity in the range of 70- 94% against seven human cancer cell lines. However, the hot water extract did not show any activity. Among the fractions of 95% and 50% ethanolic extract, significant cytotoxic activity was found in the chloroform soluble fraction of 95% ethanolic extract at 100 microg/ml; it inhibited the growth in the range of 71- 99% of seven human cancer cell lines from five different tissues viz., OVCAR- 5 (ovary), HT- 29 (colon), SK- N- MC (neuroblastoma), HEP- 2 (liver), COLO- 205 (colon), NIH- OVCAR- 3 (ovary) and A- 549 (lung). The cytotoxic activity of chloroform soluble fraction was found to be higher than 5- fluorouracil, adriamycin, mitomycin-c, and paclitaxel (anticancer drugs used as positive controls). Further in vivo studies and identification of active components from the chloroform fraction and their exact mechanism of action could be useful in designing new anticancer therapeutic agents.
• Ali, Kazi Monjur & Chatterjee, Kaushik & De, Devika & Bera, Tushar & Ghosh, Debasish. (2009). Efficacy of an aqueous extract of seed of Holarrhena antidysenterica for the management of diabetes in the experimental model rat: A correlative study with antihyperlipidemic activity. International Journal of Applied Research in Natural Products. 2. The present study reports the effects of an aqueous extract of the seed of Holarrhena antidysenterica for the management of streptozotocin (STZ) induced diabetes in rats. For the management of the carbohydrate metabolism in diabetes by the said extract, the activities of carbohydrate metabolic enzymes like glucose-6-phosphatase, glucose-6-phosphate dehydrogenase, and hexokinase in the liver along with quantification of glycogen in liver and skeletal muscle were measured, and noted a significant recovery in respect to the diabetic control group. As hyperlipidemia is one of the disorders of diabetes so, we have also assessed the serum levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDLc), very low-density lipoprotein cholesterol (VLDLc), and high-density lipoprotein cholesterol (HDLc). The said aqueous extract also resulted in a significant recovery in the levels of the abovementioned biosensors of lipid profile when treated in experimentally induced diabetic rats. The extract has no toxic effect in general which has been focused on here by the monitoring of GOT and GPT activities in the liver and kidney. The results of this study enlightened that the aqueous extract of said plant part has both antidiabetic and anti-hyperlipidemic activities.
• Jalalpure, Sunil & Bamne, Shivaji & Patil, M.B. & Shah, B. & Salahuddin, Mohammed. (2006). Anti-diabetic activity of Holarrhena antidysenterica (Linn.) Wall, bark on alloxan-induced diabetic rats. Journal of Natural Remedies. 6. 26- 30. Objective: To screen the antidiabetic potential of the bark of Holarrhena antidysenterica Linn. Materials and methods: In the present study, the bark of Holarrhena antidysenterica Linn was screened for antidiabetic activity. Bark powder of H. anti- dysenterica was subjected to hot continuous extraction (soxhlet) with various solvents like petroleum ether (40- 60°C), chloroform, butanol, butanone, and alcohol. The aqueous extract was prepared by cold maceration. After preliminary phytochemical investigation, all the extracts were evaluated for antidiabetic activity after a single dose (acute study) and after prolonged treatment (chronic study) in alloxan-induced diabetic albino rats. All the extracts were given orally at a dose of 250 mg/ kg b.w., Glibenclamide was used as the standard drug (10 mg/ kg b.w. p.o.). Results and discussion: Alcohol, butanol, chloroform, aqueous, and butanone showed significant antidiabetic activity in acute as well as prolonged treatment compared to control. Petroleum ether extract did not show significant antidiabetic activity on prolonged treatment. Among all the extracts, the alcoholic extract significantly reduced the blood glucose level after a single dose and was nearly equal to standard Glibenclamide after prolonged treatment.
• Rout, Jyoti & Sahoo, Subhalaxmi & Kanungo, Satyajit & Das, Ritarani & Sahoo, Santi. (2008). Antifungal Activities of Extracts of Holarrhena antidysenterica Wall. and Plumbago zeylanica Linn. against Aspergillus species. Plant Science Research. 30. 61- 64. The aqueous and ethanolic extracts of Holarrhena antidysenterica Wall. (Apocynaceae) and Plumbago zeylanica L. (Plumbaginaceae) were evaluated for antifungal activity against pathogenic Aspergillus species. In the present study, six Aspergillus species such as Aspergillus niger (MTCC 281), Aspergillus fumigatus (MTCC 343), Aspergillus sydowii (MTCC 635), Aspergillus nidulans (MTCC 804), Aspergillus flavus (MTCC 871) and Aspergillus versicolor (MTCC 1759) were taken against these two plant species. The in vitro antifungal activity was performed by the agar disc diffusion method. The results showed that the root extract of Plumbago zeylanica and leaf extract of Holarrhena antidysenterica exhibited the highest degree of antifungal activity towards these Aspergillus species. The ethanolic extracts of the leaf and root of these plants were more active compared to aqueous. extracts against all the investigated Aspergillus strains.
• Rajendra, Kumar & Singh, Rajendra & Head, (2018). PRE-CLINICAL TOXICITY STUDIES OF HOLARRHENA ANTIDYSENTRICA STEM BARK IN MICE AND RATS. WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES. 7. 10. 20959/ wjpps 20184- 11249. The present study has been conducted to assess the pre-Clinical safety of Holarrhena antidysentrica stem bark in mice and rat models. Albino mice (Swiss) were used in graded doses of Holarrhena antidysentrica by oral routes. Acute toxicity of Holarrhena antidysentrica showed normal behavior and no mortality up to the 14th day. The animal was treated with 2000, 1000 & 500 mg/ kg, p.o. showed dull, writhing, and 30% mortality recorded within 96 h. The effect of Holarrhena antidysentrica on hematological, biochemical, and histopathological parameters in sub-acute toxicity was carried out. Sub-acute toxicity of Holarrhena antidysentrica was found to have no significant changes in hematological, biochemical parameters, and histopathological examinations.
• Verma, Pritt & Srivastva, Sajal. (2018). Hepatoprotective effect of Ethanolic Extract of Holarrhena antidysenterica against Paracetamol Induced Toxicity in Wistar Rats. Research Journal of Pharmacy and Technology. 11. 10. 5958/ 0974- 360X. 2018. 00304. 9. Background: Holarrhena antidysenterica L. traditionally treats amoebic dysentery, diarrhea, asthma, bronchopneumonia, malaria, and some other disorders. Aim: The current study was designed to explore the effects and possible mechanisms of Holarrhena antidysenterica on paracetamol-induced hepatic damage in a rat model. Materials and methods: Hepatic damage was induced in male Wistar rats with PCM (2g kg- 1) administration for 4 weeks. A liver histopathological study was performed, and the liver function was performed by determining the (SGPT) Serum glutamate pyruvate transaminase, (SGOT) Serum glutamic oxaloacetic transaminase, Alkaline phosphatase (ALP) and Total bilirubin (TB) for evaluating the effect of HAE on hepatic damage. The possible mechanisms were investigated by measuring hepatic collagen metabolism and oxidative stress level. Findings: HA extract care significantly inhibited the loss of liver weight induced by PCM. Then the HAE improved the liver function of rats as recorded by decreased serum enzymatic activities of SGPT, SGOT ALP, and TB (p< 0.05, p< 0.01). Histopathological results indicated that HAE alleviated liver damage and reduced the formation of fibrous septa. Moreover, HAE significantly decreased liver Hydroxyproline (Hyp) content. It also decreased liver Malondialdehyde concentration, and increased activities of liver superoxide dismutase, catalase, and glutathione peroxidase. The hematological Parameter induced by HAE is normally compared with the control. The standardized extract HPTLC investigation uncovered the presence of some critical phenolic compounds. Conclusion: This examination demonstrated that Holarrhena antidysenterica could be taken as a decent common wellspring of the hepatoprotective agent.
• Ganapathy, Sujan & Ramachandra, Y L & Rai, Padmalatha. (2011). In vitro antioxidant activity of Holarrhena antidysenterica Wall. Methanolic leaf extract. Journal of Basic and clinical pharmacy. 2. 175- 8. The antioxidative potential of the methanolic leaf extract of Holarrhena antidysenterica was evaluated using hydroxyl radical, superoxide anion scavenging, and reducing power assays. The antioxidant activity of the methanol extract increased in a concentration-dependent manner. The extract showed significant reactive oxygen species (ROS) scavenging activity in all in vitro antioxidant assays and contained high levels of total phenolic content.
• Sharma, Mrinal & Singh, Navjeet. (2018). A Review of Pharmacological Aspects of Holarrhena antidysenterica. 10. 21276/saj. 2018. 7. 12. 5. Medicinal plants have been known for millennia and are highly esteemed all over the world as a rich source of therapeutic agents for the prevention of diseases and ailments. Holarrhena antidysenterica commonly known as kurci, kurchi, or kutaj has been used since ancient times. Holarrhena antidysenterica (syn. H. pubescens) belonging to the family Apocynaceae, is commended for the medicinal applications of its stem bark, leaves, and seeds in Ayurveda. In the past years, various phytochemicals have been isolated from the plant and shown traditional pharmacological activities such as analgesic, antibacterial, anti-diarrhoeal, anti-diabetic, anti-oxidant, anti-urolithic, and anti-inflammatory activities. Moreover, recent studies have shown the new activities viz. Angiotensin-converting-enzyme inhibitory, acetylcholinesterase inhibitory activity, Anti-amnesic activity, and neuroprotective activity. This review is a step to open insight into therapeutic uses for various diseases.
• The present work aims to isolate and identify new AChE inhibitors from Holarrhena anti- dysenterica. The results suggest that these alkaloids could be potential candidates for further development of new drugs against AD. Yang ZD, Duan DZ, Xue WW, Yao XJ, LI S. Steroidal alkaloids from Holarrehena anti- dysentrica as acetylcholinesterase inhibitors and the investigation for structure-activity relationships. Life Sci. 2012 Jun 14; 90 (23- 24): 929- 33. doi 10. 1016/ j. lfs. 2012. 04. 017. Epub 2012 Apr 30.
• This study was carried out to provide the pharmacological basis for the medicine of Holarrhena anti- dysenterica in gastrointestinal disorders. These results indicate that the gut stimulant and relaxant activities of Holarrhena anti- dysenterica are mediated possibly through activation of histamine receptors and Ca (++) channel blockage, respectively and this study provides sound mechanistic background for its usefulness in gut motility disorders such as constipation, colic, and possibly diarrhea. Gilani AH, Khan A, Khan AU, Bashir S, Rehman NU, Mandukhail SU. Pharmacological basis for the medicinal use of Holarrhena anti- dysenterica in gut motility disorders. Pharm Biol. 2010 Nov; 48 (11): 1240- 6. doi: 10. 3106/ 13880201003727960. Epub 2010 Sep 7.
• A new steroidal alkaloid, named anti- dysentricine, has been isolated from the seeds of Holarrhena anti- dysenterica and characterized as 3 beta-di- methylamino con- 5- in- 18- one (1). Kumar A, Ali M. A new steroidal alkaloid from the seeds of Holarrhena anti- dysenterica. Fitoterapia. 2000. Apr; 71 (2): 101- 4.
• The pharmacological and clinical application of the total as well as individual alkaloids of the plant in acute and chronic amoebic dysentery are reported (Chopra et al., 1927 & 1933; Dutta & Iyer, 1968).
• Conessine was reported to be more potent as an amoebic agent in vitro, in comparison to the other alkaloids, as tested against the ‘C’ strain of Ent. Histolytica. (Basu & Jayaswa, 1968).
• Conkurchine hydrochloride in higher doses decreased the heart rate of frogs, lowered the dog BP and dilated the rat blood vessels (Jai Shankar et al., 1961).
• The 50% ethanolic extract of root showed antiprotozoal effect against Ent. hystolytica strain STA, Trypanosoma evansi; anti-cancer effect against human epidermal carcinoma of the nasopharynx in tissue culture and hypoglycemic activity in rats.
• The fruit and stem bark extracts also revealed an anti-spasmodic effect on isolated guinea pig ileum (Dhar et al., 1968).
• The stem bark showed hypotensive action. The LD50 of the ethanolic extract of the fruit was 250 mg/kg i.p., whereas that of stem bark was 1000 mg/kg orally in mice (Dhar et al., 1968).
• The mother tincture prepared from the bark with 70% alcohol exerted the maximum antispasmodic activity as compared to the other tinctures prepared with varying percentages of alcohol (Nandi & Mazumdar, 1979).
• A clinical study (on the side effects) in 11 patients revealed that the drug can lead to subjective symptoms, as well as hypotension (Chaturvedi & Singh, 1983).

Recent Research on W. tinctoria

• 3- Orhamnoglucoside from the flowers exhibited significant anti-inflammatory activity in the rats against carrageenin-induced rat paw edema (Sethuraman et al., 1984).
• Jain, Pritam & Bari, Sanjaykumar & Surana, Sanjay. (2011). Acute Oral Toxicity of Abelmoschus manihot and Wrightia tinctoria in Mice. Pharmacognosy Journal. 3. 78– 81. 10. 5530/jp. 2011. 25. 14. Abelmoschus manihot and Wrightia tinctoria, belonging to the botanical family Malvaceae and Apocynaceae, have been traditionally used by the locals in India for the treatment of various ailments. The current study reports the outcome of an acute oral toxicity investigation of Abelmoschus manihot and Wrightia tinctoria, on ICR mice. No mortalities or evidence of adverse effects have been observed in ICR mice following acute oral administration at the highest dose of 2500 mg/ kg crude extracts of Abelmoschus manihot and Wrightia tinctoria. This is the first report on the acute oral toxicity of Abelmoschus manihot and Wrightia tinctoria and the findings of this study agree with those of in vitro experiments and thus provide scientific validation on the use of the leaves of Abelmoschus manihot and Wrightia tinctoria.
• Wrightiadione exhibited cytotoxicity against murine P-388 lymphocytic leukemia cell line (ED50, 1.1. ug/ ml)- (Phytochem. 1992, 31, 4333).
• Fatima, Nishat & Ahmad, Mohammad & Ansari, Jamal & Ali, Zulfiqar & Khan, Abdul & Mahdi, Abbas Ali. (2016). Anticancer, antioxidant potential and profiling of polyphenolic compounds of Wrightia tinctoria Roxb. (R. Br.) bark. Journal of Advanced Pharmaceutical Technology & Research. 7. 10. 4103/ 2231- 4040. 191428. Anticancer, antioxidant potential and profiling of polyphenolic compounds of Wrightia tinctoria Roxb. (R. Br.) bark Abstract Wrightia tinctoria Roxb. (R.Br.) is an Ayurvedic remedy, ethnomedical used in the treatment of various ailments. The present work was carried out to evaluate the anticancer and antioxidant activity as well as total phenolic and phytochemical contents of W. tinctoria bark methanolic extract (WTBM) by high‑performance liquid chromatography (HPLC)‑diode array detector. Antiproliferative activity of WTBM was evaluated against MDA‑ MB‑ 231 and MCF‑ 7 cancer cells by 3‑ (4, 5‑ dimethylthiazol‑ 2‑ yl) ‑2, 5‑ diphenyltetrazolium bromide assay, colony formation, and Hoechst staining. In addition, the antioxidant potential was determined by 2, 2‑ diphenyl‑ 1‑ picrylhydrazyl (DPPH) radical scavenging activity and 2, 2‑ and‑ bis‑ 3‑ ethylbenzothiazoline‑ 6‑ sulfonic acid (ABTS) radical cation decolorization assay. Total phenolic content was assessed by the Folin–Ciocalteu method. The results demonstrated that WTBM exhibited a significant antiproliferative effect against MDA‑ MB‑ 231 (IC 50 = 88.9 ± 1.27 µg / ml) and MCF‑ 7 (IC 50 = 45.71 ± 7.74 µg/ ml) cancer cells in a time‑ and dose‑dependent manner. WTBM significantly suppresses colony formation and induces apoptosis in both MDA‑ MB‑ 231 and MCF‑ 7 cells as evidenced by morphological assessment, clonogenic assay, and Hoechst staining. The total phenolic content of WTBM was found to be 30.3 gallic acid equivalent mg/g dry weight of bark extract while IC 50 value for DPPH and ABTS radical scavenging activity was 72.2 ± 2.8 µg/ml and 45.16 ± 1.95 µg/ml, respectively. HPLC analysis showed the presence of gallic acid, rutin, and quercetin in WTBM. These findings demonstrated that WTBM significantly inhibited the proliferation of breast cancer cells and induced apoptosis, suggesting the potential chemo-preventive activity of W. tinctoria bark.
• Tharkar, Priyanka & Tatiya, A & Surana, Sanjay & Bhajipale, N & Deore, S. (2009). Anti-inflammatory Study of Wrightia tinctoria R. Br Stem Bark in Experimental Animal Models. Int J Pharm Tech Res. In the present study, the bark of Wrightia tinctoria was investigated for anti-inflammatory activity by carrageenan-induced rat paw edema and cotton pellet-induced granuloma method. The various extracts showed inhibition of rat paw edema and percent granuloma changes at the dose of 200 mg/ kg when compared to the control group. The activity was compared with that of the standard drug diclofenac sodium (13.5 mg/ kg /b w, p.o) Keywords: Wrightia tinctoria, granuloma method, anti-inflammatory activity. INTRODUCTION In the traditional system of ayurvedic medicines, plant products either single drug or in combination with others are considered to be less toxic and free from side- effects as compared to synthetic drugs. Inflammatory diseases are the major cause of morbidity in the working force throughout the world. Many drugs produced a dramatic symptomatic improvement in rheumatic processes, but all of them shared common side effects like gastrointestinal irritation. 2. Wrightia tinctoria R. Br (Apocynaceae) bark commonly known as kalakuda, indrajau (in Marathi). It is a small deciduous tree, generally up to 7.5 m in height, found in Rajasthan, Madhya Pradesh, and the peninsular region of India. The bark is light grey, scaly, and smooth 3. It is used in dropsy and bilious affections 4, 5. A literature survey revealed that no scientific investigation regarding the anti-inflammatory activity of the bark was mentioned to evaluate scientifically the potency of various extracts of Wrightia tinctoria based on its ethnobotanical clue.
• Dhanabal, S.P. & Raj, Baskar & N, Muruganantham & Krishnamurthy, Praveen & Raghu, P.S. (2012). Screening of Wrightia tinctoria leaves for Anti psoriatic activity. Hygeia. 4. 73-78. Plan: The hydroalcoholic extract of Wrightia tinctoria leaves was evaluated for anti-psoriatic activity by mouse tail test. Methodology: Anti-psoriatic activity was performed at a dose of 200 mg/kg body weight in mice (25- 30 g). Isoretinoic acid (0.5 mg/ kg) was used as the standard. The degree of orthokeratosis, drug activity, and the relative epidermal thicknesses were calculated and statistically analyzed. The extract was also evaluated for its antioxidant potential by DPPH, nitric oxide, and hydrogen peroxide radical scavenging assays. Outcome: The extract produced a significant (p< 0.01) degree of orthokeratosis compared to control and the drug activity was found to be 70.18%, which is more potent than the standard (57. 43%). The extract showed prominent antioxidant activity in all the assays. The present study concludes that the selected plant has anti-psoriatic activity and can be used for psoriasis treatment.
• Thabah, P. & Gopal, Murugananthan & Joshi, N.C. & Nandakumar, K & Lakshman, Kuruba & Talwar, S. (2009). Immunomodulatory activities of Wrightia tinctoria (Roxb.) R. Br Bark extract. Pharmacologyonline. 3. 663-669. Wrightia tinctoria (Roxb.) R.Br. (WT) has been reported to exhibit several therapeutic uses such as astringent, stomachic, febrifuge, skin diseases, and tonic in India. The objective of the present study was to investigate the immunomodulatory activity of the bark extracts of WT using delayed type hypersensitivity reaction and carbon clearance assay. Powdered dried barks of WT were extracted with petroleum ether 60 – 80°C (PEWT), alcohol (ALWT), and aqueous alcohol (AQWT) (60 % water + 40 % ethanol) successively. PEWT and ALWT extracts (200, 400 mg/ kg, p. o) produced a significant increase in delayed-type hypersensitivity in response to sheep red blood cells (SRBC). PEWT showed better activity than ALWT in delayed-type hypersensitivity response. ALWT (200 and 400 mg/ kg, p. o) in a dose-dependent manner has shown a significant increase in phagocytic activity. The above results reveal that ALWT possesses immunostimulant activity in carbon clearance assay whereas PEWT and ALWT showed immunomodulatory activity in the delayed-type hypersensitivity model.
• Bigoniya, Papiya & Rana, A.C. (2009). Antidiarrheal and antispasmodic activity of Wrightia tinctoria bark and its steroidal alkaloid fraction. Pharmacologyonline. 3. 298- 310. Wrightia tinctoria Roxb. R.Br. (family Apocynaceae) bark ethanol extract and an isolated steroidal alkaloid fraction (WTSA) were investigated on different experimentally induced diarrhea models of rats, isolated rat ileum, and on enteric bacterium to establish the therapeutic potential. The extract at 500 and 1000 mg/kg dose and WTSA at 50 and 100 mg/kg dose significantly inhibited the frequency and wetness of fecal droppings in castor oil-induced diarrhea. Extract and WTSA decreased propulsion of charcoal meal and also reduced prostaglandin E2-induced enter pooling. WTSA reduced the amplitude, frequency, and tone of spontaneous gut movement. Alkaloid fraction also inhibited acetylcholine (Ach)-induced contraction of rat ileum. W. tinctoria alkaloid has antisecretory, spasmolytic, anti- enteropooling, antimotility, and anti-peristaltic activity. These results substantiate good antidiarrheal activity of W. tinctoria against secretory, osmotic, motility-related, and inflammatory diarrhea.
• Reddy, Y & Venkatesh, Sama & Ravichandran, T & Murugan, V & Suresh, Bhavana. (2002). Antinociceptive activity of Wrightia tinctoria bark. Fitoterapia. 73. 421-3. 10. 1016/ S0367- 326X (02) 00126- 0. The ethyl acetate, acetone and methanol extracts of Wrightia tinctoria bark showed antinociceptive activity on acetic acid-induced writhing test in mice, their effects being comparable to that of acetylsalicylic acid.
• Rajalakshmi, G.R. & Harindran, Jyoti. (2013). In vitro anthelmintic activity of Wrightia tinctoria. International Journal of PharmTech Research. 5. 308-310. Alcoholic and aqueous extracts from leaves of Wrightia tinctoria were investigated for their anthelmintic activity against Raillietina spiralis and Ascaridia galli. The three concentrations (10, 25, and 50mg/ml) of extract were studied in an activity that involved the determination of the time of paralysis and time of death of the worm. The extract exhibited significant dose-dependent anthelmintic activity. Piperazine citrate was included as a standard reference and distilled water as a control.
• Dixit, Ashish & Jain, A & Sharma, Naveen & Saluja, Gurudeep & Acharya, Manutosh & Rathore, Dheerendra & Joshi, Ramakant & Sharma, Pankaj & Singh, Yogendra & Asst, Ashish. (2015). HEPATOPROTECTIVE ACTIVITY OF WRIGHTIA TINCTORIA LEAVES AGAINST D- GALACTOSAMINE INDUCED HEPATOTOXICITY IN RATS. 5. 422. The present study was conducted to evaluate the hepatoprotective activity of phytoconstituents of methanolic extract of leaves of Wrightia tinctoria against D- galactosamine-induced liver damage in albino rats. The phytoconstituents (100 mg/kg) were administered orally to the animals with hepatotoxicity-induced D-galactosamine (400 mg/kg for two days). Silymarin (50 mg/ kg) was given as a standard hepatoprotective drug. All the test drugs were administered orally in the suspension form to the animals. The combination of silymarin and methanolic extract of Wrightia tinctoria leaves was also given to animals to compare with individual groups as well as toxicant groups. The combination of silymarin and methanolic extract of Wrightia tinctoria leaves was effective in protecting the liver against the injury induced by D- galactosamine in rats. This was evident from a significant reduction in serum enzyme alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and total bilirubin. These biochemical observations were also supplemented by Histopathological observations of livers sections. It was concluded from the results that the combination of silymarin and methanolic extract of Wrightia tinctoria leaves reduces the hepatoprotective activity against D-galactosamine-induced hepatotoxicity in rats.
• Selvam, Pakia & Murugesh, N & Witvrouw, M & Keyaerts, Els & Neyts, Johan. (2009). Studies of Antiviral Activity and Cytotoxicity of Wrightia tinctoria and Morinda citrifolia. Indian Journal of pharmaceutical sciences. 71. 670-2. 10.4103/0250-474X.59550. Different extracts of leaf parts of Wrightia tinctoria and fruit powder of Morinda citrifolia have been studied against replication of HIV-1 (IIIB) in MT- 4 cells and HCV in Huh 5.2 cells. The chloroform extract of Wrightia tinctoria exhibited a maximum protection of 48% against the cytopathic effect of HIV-1(IIIB) in MT-4 cells. Fruit juice of Morinda citrifolia exhibited a displayed marked cytotoxic activity in lymphocyte (MT- 4) cells (CC50: 0.19 mg/ml). The 50% effective concentration for inhibition of HCV subgenomic replicon replication in Huh 5- 2 cells by Morinda citrifolia was 0.98 mug/ ml and by chloroform extract of Wrightia tinctoria was 10 mug/ml. The concentration that reduced the growth of exponentially proliferating Huh 5-2 cells by 50% was greater than 50 mug/ml.
• Sathyanarayanan, S & Selvam, Pakia & Jose, Asha & George, Rijo & Revikumar, K & Neyts, Johan. (2008). Preliminary phytochemical screening and study of anti-viral activity and cytotoxicity of Wrightia tinctoria. Int J Chem Sci. 7. Wrightia tinctoria was investigated for the preliminary phytochemical analysis and characterization by various instrumental techniques. Indole derivatives such as isatin, induribine, tryphanthrine, and fatty acids were identified. Methanolic extract of leaf parts of Wrightia tinctoria (WT) has been studied against replication of HCV in Huh 5.2 cells. The 50% effective concentration for inhibition of HCV in RNA subgenomic replicon replication in huh 5-2 cells (luciferase assay) by CWT was found to be 15 The concentration that reduced the growth of exponentially proliferating Huh 5-2 cells by 50% was greater than 50 /mL.
• Vedhanarayanan, P. & P, Unnikannan & Sundaramoorthy, Prem. (2013). Antimicrobial activity and phytochemical screening of Wrightia tinctoria (Roxb) R. Br. 2. 123-125. In the present investigation, the antibacterial activity of different extracts (Chloroform, ethanol, and methanol) of Wrightia tinctoria has been studied against the human pathogenic bacterial strains, Escherichia coli, Bacillus subtilis, Staphylococcus aureus, and Pseudomonas aeruginosa by disc diffusion method on agar. The findings showed potential antibacterial properties of the extracts against the organisms tested. Among the three solvents tested, ethanol extract of leaf showed a higher inhibition zone. Ethanol extract of Wrightia tinctoria exhibits maximum zone of inhibition against Escherichia coli (29 mm), Bacillus subtilis (24 mm) Staphylococcus aureus (30 mm) and Pseudomonas aeruginosa (24 mm). Preliminary phytochemical analysis of Wrightia tinctoria showed the presence of alkaloids, flavonoids, phenols, saponins, steroids, and tannins.
• Shruthi, A. & Latha, K.P. & Vagdevi, H.M. & Vaidya, V.P. & Pushpa, B. & Shwetha, C. (2010). In vitro anthelmintic activity of leaves extract of Wrightia tinctoria. 2. 2043-2045. The development of anthelmintic resistance and the high cost of conventional anthelmintic drugs led to the evaluation of medicinal plants as an alternative source of anthelmintics. The present study aimed to determine the anthelmintic activity of crude petroleum ether and chloroform extracts of leaves of Wrightia tinctoria using Pheretima Posthuma. Three concentrations (2.5, 5.0, 7.5 mg/ ml) of each extract were studied in the activity, which involved the determination of the time of paralysis and time of death of the worms. Piperazine citrate is used as a standard reference and normal saline as a control. The present study proves the potential usefulness of leaves of Wrightia tinctoria as a comparable anthelmintic agent.

Recent Research on W. tomentosa

• Nagarajan, Kandasamy & Mazumder, Avijit & Ghosh, L. (2008). Toxicological Evaluation and Antinociceptive Effects of Wrightia tomentosa In Mice. Nigerian Journal of Natural Products and Medicine. 11. 10. 4314/ njnpm. v11i1. 11885. Acute toxicity testing and the antinociceptive activity in mice of ethanolic extract of Wrightia tomentosa leaves and bark were carried out. The bark extract (400 mg/ kg) exhibited maximum antinociceptive action after 120 minutes of administration with a value of 14.67 ± 4.17 seconds in comparison with standard aspirin having a reaction time of 8.33 ± 1.72 seconds after 120 minutes. The leaf extract was not as active.
• Nagarajan, Kandasamy & Mazumder, Avijit & Ghosh, L.K. (2007). Comparative anti-allodynic effects and toxicity studies for the herbal Wrightia tomentosa leaf & bark in Swiss albino mice. Pharmacology online. 3. 294- 307. Ethanol bark and leaf extract of Wrightia tomentosa have been shown to exhibit anti-allodynic activity in Swiss albino mice. This study aimed to evaluate the comparative anti-allodynic effects of both the leaf and bark extract of the plant along with an acute oral toxicity study. Acute toxicity studies were carried out by using the Organization of Economic Co-operation and Development (OECD) guidelines 423, using a minimum number of male Wistar rats with defined doses (5, 50, 300, 2000 mg/ kg body weight) of extracts. The extracts treated animals did not show any mortality or observable signs of toxicity. From the toxicity studies, 1/ 5th of the dose was selected as the maximum dose for testing of anti-allodynic activity. The comparative anti-allodynic activity of ethanolic leaf and bark extracts was determined, with six groups of six albino mice each by tail immersion method. After the administration of test drugs in two different doses, the reaction time was measured at 0, 30, 60, 120, and 180 minutes. The highest reaction time was considered as a maximum anti-allodynic effect, which was exhibited by the bark extract (400 mg/ kg) after 120 minutes of administration with a value of 14.67 ± 4.17 seconds in comparison with standard drug aspirin (8.33 ± 1.72). The current studies generally focus on induced pain in animal models that are designed to evaluate anti-allodynic effects for humans shortly.
• David, Ernest & Elumalai, Kuppusamy & Therasa, S.Viviyan & Thirunavukkarasu, Dr. Thirumalai. (2010). Evaluation of the anti-inflammatory activity of the leaf extract of Wrightia tomentosa. Roem & Schult. Journal of Pharmacy Research. 3. The methanol: dichloromethane (1:1) extract of the leaves of Wrightia tomentosa (Family: Apocynaceae), a traditional medicine plant, was tested for anti-inflammatory activity dextran-induced paw edema in rats. The results revealed that methanol: dichloromethane extract significantly ameliorates dextran-induced edema. Acetylsalicylic acid was used as a reference drug.
• Chakravarti, Bandana & Maurya, Ranjani & Siddiqui, Jawed & Bid, Hemant & Rajendran, S.M. & Yadav, Prem Prakash & Konwar, Rituraj. (2012). In vitro anti-breast cancer activity of ethanolic extract of Wrightia tomentosa: Role of pro-apoptotic effects of oleanolic acid and ursolic acid. Journal of Ethnopharmacology. 142. 72- 9. 10. 1016/ j. jep. 2012. 04. 015. Wrightia tomentosa Roem. & Schult. (Apocynaceae) is known in traditional medicine for anti-cancer activity along with other broad indications like snake and scorpion bites, renal complications, menstrual disorders, etc. However, the anti-cancer activity of this plant or its constituents has never been studied systematically in any cancer type so far.
• To evaluate the anti-cancer activities of the ethanolic extract of W. tomentosa and identified constituent active molecule(s) against breast cancer.
• Powdered leaves of W. tomentosa were extracted with ethanol. The ethanolic extract, subsequent hexane fractions, and fraction F- 4 of W. tomentosa was tested for its anti-proliferative and pro-apoptotic effects in breast cancer cells MCF- 7 and MDA – MB- 231.
• The ethanolic extract, subsequent hexane fractions, and fraction F- 4 of W. tomentosa inhibited the proliferation of human breast cancer cell lines, MCF- 7 and MDA- MB- 231. The fraction F- 4 obtained from hexane fraction inhibited proliferation of MCF- 7 and MDA- MB- 231 cells in a concentration and time-dependent manner with IC₅₀ of 50 μg/ ml and 30 μg/ ml for 24 h, 28 μg/ ml, and 22 μg/ ml for 48 h and 25 μg/ ml and 20 μg/ ml for 72 h respectively. The fraction F- 4 induced G1 cell cycle arrest, reactive oxygen species (ROS) generation, loss of mitochondrial membrane potential, and subsequent apoptosis. Apoptosis is indicated in terms of increased Bax/ Bcl- 2 ratios, enhanced Annexin-V positivity, caspase 8 activations, and DNA fragmentation. The active molecule isolated from fraction F- 4, oleanolic acid, and urosolic acid inhibited cell proliferation of MCF- 7 and MDA- MB- 231 cells at IC₅₀ value of 7.5 μM and 7.0 μM respectively, whereas there is devoid of significant cell inhibiting activity in non-cancer originated cells, HEK- 293. In both MCF- 7 and MDA- MB-231, oleanolic acid, and ursolic acid-induced cell cycle arrest and apoptosis as indicated by the significant increase in Annexin-V positive apoptotic cell counts. Our results suggest that W. tomentosa extracts have significant anti-cancer activity against breast cancer cells due to the induction of the apoptosis pathway. Olenolic and ursolic acids are important constituent molecules in the extract responsible for the anti-cancer activity of W. tomentosa.
• Nagarajan, Kandasamy & Mazumder, Avijit & Ghosh, L.K. (2008). In vitro antioxidant activity of alcoholic extracts of Wrightia tomentosa. Pharmacologyonline. 1. 196- 203. The objective of the present investigation is to assess the most potent antioxidant alcoholic extract from the leaf & bark of Wrightia tomentosa. The invitro-methods used to-predict the antioxidant effect was DPPH radical scavenging assay, reducing power ability, Phospho molybdate method & the estimation of total phenolic and flavonoid contents using pyrocatechol & quercetin as standards. The % scavenging activity at different concentrations was determined and the IC 50 value of the extracts was compared with that of standard, ascorbic acid. The ethanolic bark extract gave an IC 50 value of 75.0 μg/ml when compared with leaf extract (IC 50 = 135.1 μg/ml). The reducing power was investigated by Fe3+ – Fe2+ transformation in the presence of extracts tested using Butylated hydroxytoluene as standard. The ethanol bark extract showed the highest reducing ability having an absorbance of 0.690 using 800 μg/ ml concentration at 700nm. The total antioxidant capacity by the phosphor molybdate method is expressed as α tocopherol equivalents. Among the extracts tested, the ethanol bark extract contains 8.3 μg Vitamin E equivalent/ mg, which has appeared twice greater antioxidant capacity than the leaf extract (4.2 μg Vitamin E equivalent/mg). The content of total phenolics (9.1 μg Pyro- catechol equivalent/mg) and total flavonoids (20.0 μg Quercetin equivalent/mg) in the ethanol bark extract was considerably higher than in the leaf extract. Based on the above results, the higher the flavonoid content, the higher the antioxidant capacity was very well observed with ethanolic bark extract. Hence the alcoholic extract of Wrightia tomentosa dried bark could be considered for the preparation of nutraceuticals with potent antioxidant effects suitable for the prevention of human disease.
• Shruthi, A. & Latha, K.P. & Vagdevi, H.M. & Pushpa, B. & Shwetha, C.. (2012). Anti-diabetic activity of the leaf extracts of Wrightia tinctoria on alloxan-induced diabetic rats. Journal of Chemical and Pharmaceutical Research. 4. 3125- 3128. In the present study, the investigation has been carried out to evaluate the effect of the different extracts of the leaves of Wrightia tinctoria on alloxan-induced diabetic rats of the Wistar strain. The experiment was carried out using six groups of albino rats. Chloroform extract showed significant anti-diabetic activity when compared to the standard drug glibenclamide.
• Sawale, Jyotiram & Panchal, Chandrawadan & Padmane, Suhas & Poul, B & Patel, Jay. (2014). IN VITRO ANTIOXIDANT AND ANTI-INFLAMMATORY ACTIVITY OF WRIGHTIA TINCTORIA LEAVES. WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES. 3. 964- 972. Background: Since free radicals play such an important role in the disease scenario of an individual, a thorough understanding of the various physiologically significant free radicals is of paramount importance before the search for the radical scavengers or the antioxidant principles to treat the physiological disorders caused by them such as inflammation. Objective: To explore newer antioxidant and anti-inflammatory agents from natural sources. Material and Methods: The antioxidant activity of the methanolic extract of Wrightia tinctoria leaves was determined by measuring its 1,1 diphenyl- 2- pycryl hydroxyl (DPPH) radical scavenging activity, reducing power and ability to inhibit lipid peroxidation. Anti-inflammatory activity was evaluated by using a carrageenan-induced rat paw edema model in healthy Wistar albino rats. Results and Discussion: Methanol extract exhibited strong total antioxidant activity. Concentrations of 50, 100, 200, 400, 800 and 1000μg /mL showed 52.34, 53.69, 53.69, 56.37, 58.60, and 61.07% inhibition of lipid peroxidation respectively. In addition, methanol extract had effective reducing power, and DPPH free radical scavenging activity at 400 μg/ mL, the same extract was also evaluated for its anti-inflammatory activity at doses 100, 200, and 400 mg/ kg. Conclusion: Methanolic extract showed predominantly significant activity in a dose-dependent manner, which is comparable to reference standard diclofenac sodium. Phytochemical studies indicated the presence of alkaloids and phenols are the major components in the extract and it may be further concluded that these constituents may be responsible for antioxidant and anti-inflammatory activity.