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Chemical Evaluation of Mucuna Species for L-Dopa Content - an Anti-Parkinson’S Drug Yielding Medicinal Plant from India

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Chemical Evaluation of Mucuna Species for L-Dopa Content - an Anti-Parkinson’S Drug Yielding Medicinal Plant from India Indian Journal of Traditional Knowledge Vol. 17(1), January 2018, pp. 148-154 Chemical evaluation of Mucuna species for L-dopa content - an anti-Parkinson’s drug yielding medicinal plant from India Archana P Raina1* & RC Misra2 1Division of Germplasm Evaluation, ICAR - National Bureau of Plant Genetic Resources, New Delhi-110 012, India; 2ICAR - National Bureau of Plant Genetic Resources, Regional Station, Cuttack-753 006, Odisha, India E-mail: [email protected] Received 23 November 2016, revised 2 June 2017 Mucuna species is an important medicinal plant of India which is the best known natural source of bioactive compound L-dopa (L-3,4-dihydroxy phenylalanine) used as potential drug for the treatment of Parkinson’s disease. The present study was undertaken for the chemical evaluation of four species of Mucuna, viz.; M. pruriens var. pruriens, M. pruriens var. utilis (Wall. ex Wight) L. H. Bailey, M. monosperma Wight, M. nigricans (Lour.) Steud., M. gigantea (Willd.) DC., collected from wild habitats of Odisha state of eastern India for pharmaceutically valued compound L-dopa. Results indicated a wide range of germplasm variability in seed size, color and L-dopa content. Among the four Mucuna species, highest L-dopa content was found in M. pruriens var. pruriens germplasm varying between 4.91 % - 7.09 %. Other Mucuna species having high L-dopa were M. gigantea (6.76 %), M. nigricans (6.16 %) and M. monosperma (4.61 %); while M. pruriens var. utilis showed least L-dopa content of 1.22 %. Promising accessions with high L-dopa content (> 6 %) were IC599290, IC599342, IC599336, IC599361 and IC599350. Superior accessions along with other rarely occurring Mucuna species can be exploited for large-scale cultivation of this drug plant for pharmaceutical use. Keywords: Mucuna species, Germplasm, HPTLC, L-Dopa, Mucuna pruriens, Parkinson’s disease IPC Int. Cl.8: A61K 36/00, A61K 38/00, A01D 8/46, A01D 8/00 Mucuna is a well-known medicinal and underutilised popularly known as Velvet bean, cowhage, kewanch legume plant which is widely exploited for its seed and atmagupta. All parts of Mucuna plant are pods. The genus Mucuna belongs to the family reported to possess useful phytochemicals of high Fabaceae and has approximately 150 species reported medicinal value for human and veterinary importance from worldwide1. The plant is widely distributed and also constitute an important raw material in throughout the India and in other parts of the tropics Ayurvedic and folk medicines. It is a constituent of including Central and South America. In India, 14 more than 200 indigenous drug formulations. This species of Mucuna are found in the foothills of the plant is pharmacologically important for various Himalayas, the plains of West Bengal, Madhya activities reported like antidiabetic, aphrodisiac, Pradesh, Karnataka, Kerala, Andra Pradesh, Uttar antineoplastic, antiepileptic, antimicrobial activities, Pradesh and the Andaman and Nicobar islands2. etc4. Mucuna is an unconventional plant species Out of these, M. pruriens, M. monosperma and having promising nutritional, pharmaceutical and M. gigantea are widely distributed all over; while bioactive constituents. Mucuna seeds are the best M. nigricans is distributed mainly in the North- nutritional source due to high content of protein, Eastern parts of India. Mucuna pruriens (L.) DC. is carbohydrate, lipids, fiber and minerals5. Researchers widely used in traditional Ayurvedic system of Indian from various countries have also identified Mucuna as medicine for the management of male fertility, a good nutritional supplement in livestock feed, as a nervous disorders and as an aphrodisiac3. It is fodder crop. The wild species of M. pruriens var. —————— pruriens and M. pruriens var. hirsuta are being *Corresponding author cultivated in India and Brazil as a cover and green RAINA & MISRA: CHEMICAL EVALUATION OF MUCUNA SPECIES FOR L-DOPA CONTENT FROM INDIA 149 manure crop. The demand for Mucuna is increasing in raw seed where it ranges from 4.47 % - 5.39 %12. day by day due to its pharmaceutical potency6. A wide range of variation in L-dopa content of Mucuna species have been known to have M. pruriens seeds have been reported by earlier bioactive compound L-dopa (L-3,4-dihydroxy phenyl workers6,7,11,13. However, little is known about alanine), a non-protein amino acid which is a the production of L-dopa in other species of precursor of the neurotransmitter dopamine7. It is Mucuna except for few reports on M. monosperma14,15, being used in the treatment of Parkinson’s disease, M. gigantea16, M. sanjappae17 and M. utilis18,20. a degenerative disease of the nervous system Keeping in view the increasing demand for herbal characterized by muscular rigidity, difficulty with drugs, importance of this genus and vast diversity of balance and walking, depressions and dementia. Mucuna germplasm available in Indian continent, L-Dopa, a dopamine precursor, either alone or efforts were made to collect diverse germplasm of in combination with aromatic amino acid Mucuna species from wild habitats of the state of decarboxylase inhibitor is the most effective drug for Odisha, India. These accessions have been conserved the treatment of Parkinson’s disease, since dopamine in the National Seed Genebank at ICAR-NBPGR, fails to pass through the blood brain barrier8. Use of New Delhi, India. The role of genetic resource biological sources for production of L-dopa, is always characterization in plant breeding endeavours is well desirable and advantageous because the chemical recognized. However, despite presence of rich synthesis results in racemic DL-mixture, which is genepool, such options are explored to minimal in inactive and furthermore separation of enantio- Mucuna species, problably due to lack of data on merically pure L-dopa from this mixture, is very germplasm attributes. Therefore, there is a need to difficult and cumbersome. In addition, D-dopa undertake detailed characterization of Mucuna genetic interferes with the activity of dopa decarboxylase, the resources in India to reinforce the local crop enzyme involved in the production of dopamine in the impovement program. In view of this, the present brain. L-dopa isolated from Mucuna was found to be study was undertaken with the main objective to more effective than the synthetic product9. The plant screen Mucuna species germplasm collected from sources have several advantages over the microbial different regions of Odisha state of India for L-dopa and chemically synthesized L-dopa, viz. it is natural, content and to identify high L-dopa yielding lines inexpensive source and provides additional benefits which can be used in future breeding programs and like antioxidant properties to reduce oxidative stress for pharmaceutical utilization. Four species of produced during Parkinson’s disease9,10. In a Mucuna namely, M. monosperma, M. nigricans, M. screening survey of more than 1000 species in 135 gigantea, M. pruriens var. pruriens and M. pruriens plant families, Mucuna was found as the only species var. utilis collected from wild habitats of Odisha from with sufficient L-dopa to suggest a possible use for its eastern India were analyzed for the presence of L- commercial production11. Various L-dopa yielding dopa. These plant species will be a potential candidate plant species such as M. holtonii, M. monosperma, M. for extraction of L-dopa and will be a good addition utilis, M. gigantea and M. pruriens were found to be a to the existing natural sources of anti-Parkinson’s promising source of L-dopa7. Demand for Mucuna drug since demand for L-dopa is continuously seeds for L-dopa is constantly increasing in the escalating. international drug market. M. pruriens has been Materials and methods profitably exploited for the extraction of this drug since long time12. Large-scale extraction of L-dopa Plant material from the wild populations of this plant has led to its Thirty accessions belonging to four species of limited availability in natural conditions. There is an Mucuna were collected by selective sampling urgent need for collection of diverse germplasm of method from wild regions of 10 districts of Odisha in Mucuna species from wild sources to identify superior India through explorations. Authentication and types and other species of Mucuna for exploitation for identification of collected Mucuna species was done anti-Parkinson’s drug. This will also ease the burden by Dr. R.C. Misra, Taxonomist at ICAR-NBPGR, on M. pruriens for L-dopa extraction. Distribution of Cuttack and voucher samples were preserved in the L-dopa in the species showed wide variation in herberaium. These germplasm collections comprised different parts of the plant with highest concentration of 26 accessions of M. pruriens var. pruriens (Kewanch) 150 INDIAN J TRADIT KNOWLE, VOL. 17, NO. 1, JANUARY 2018 and one accession each of M. pruriens var. utilis (20 cm × 20 cm) were purchased from E. Merck (Wall. Ex Wight) L. H. Bailey, M. monosperma (Darmstadt, Germany). A stock solution of L-dopa Wight (Negro bean), M. nigricans (Lour.) Steud. (1mg/mL) was prepared by dissolving an accurately (Black Jade vine) and M. gigantea (Willd.) DC. (Sea weighed 10 mg of L-dopa standard in 10 mL of 0.1N bean or Elephant Cowitch). The details of NBPGR HCl in a volumetric flask. Standard working solution accessions along with place of collection are given in was prepared by diluting it with 0.1 N HCl to make Table 1. These germplasm accessions were grown final concentration of 0.1 µg/µL. under uniform environmental conditions at NBPGR Regional Station, Cuttack, Odisha for evaluation Sample preparation during the years 2013-14 and 2014-15. Superior Matured seeds were harvested and dried at 60 °C. accessions were further evaluated and validated for Seeds were powdered to a mesh size of 60 after high L-dopa content in subsequent years. removing seed coats. Seed extract was prepared by refluxing 0.5 g of seed powder with 50 mL 0.1N HCl Chemicals on a boiling water bath for 30 min.
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