title: Validation of ethnomedicinal potential of Tinospora cordifolia for anticancer and immunomodulatory activities and quantification of bioactive molecules by HPTLC.
abstract: BACKGROUND
  Tinospora cordifolia (Willd.) Miers ex Hook. f. & Thomas. (Menispermaceae) is one of the most widely used plants in various traditional medicinal systems including "Ayurveda". The plant is used for the treatment of jaundice, rheumatism, urinary disorder, skin diseases, diabetes and anemia. The phytoconstituents present in the plant belongs to different class of compounds such as alkaloids, diterpenoids lactones, glycosides, steroids, phenol, aliphatic compounds and polysaccharides.
  
  
  OBJECTIVE
  The aim of present study was the isolation, structure elucidation, quantification and pharmacological evaluation of secondary metabolites from T. cordifolia for anticancer and immunomodulatory activities.
  
  
  METHODS
  Different extracts and fractions were prepared from the stem of T. cordifolia. Pure molecules were isolated using normal phase chromatography and characterized on the basis of NMR and mass spectroscopic techniques. The anti-cancer and immunomodulatory activities of different extracts, fractions and isolated compounds were evaluated against four different human cancer cell lines, KB (human oral squamous carcinoma), CHOK-1 (hamster ovary), HT-29 (human colon cancer) and SiHa (human cervical cancer) and murine primary cells respectively. A simple, normal phase HPTLC method was also developed for the quantification of three bioactive compounds i.e N-formylannonain (1), 11-hydroxymustakone (5) and yangambin (8) in the stem of T. cordifolia hosted on fifteen different plants.
  
  
  RESULTS
  Chromatographic purification of different fractions led to the isolation of eight pure molecules i.e N-formylannonain (1), magnoflorine (2), jatrorrhizine (3) palmatine (4), 11-hydroxymustakone (5), cordifolioside A (6), tinocordiside (7) and yangambin (8). All extracts and fractions were active against KB and CHOK-1 cells whereas among the pure molecules palmatine (4) was found to be active against KB and HT-29; tinocordiside (7) against KB and CHOK-1; yangambin (8) against KB cells however N-formylannonain (1) and 11-hydroxymustakone (5), was found active for immunomodulatory activity. HPTLC quantification of three active molecules i.e N-formylannonain (1), 11-hydroxymustakone (5), and yangambin (8) were found in highest quantity in the stem of T. cordifolia hosted on Mangifera indica, however, other two active molecules were not quantified due to their insufficient quantity.
  
  
  CONCLUSIONS
  Eight compounds have been isolated and characterized belonging to different classes. The pharmacological evaluation of extract, fractions and pure molecules revealed the ethnomedicinal value of T. cordifolia for anticancer and immunomodulatory activities.
authors.affiliation: Academy of Scientific and Innovative Research, New Delhi, India; Natural Product Chemistry and Process Development Division, CSIR-Institute of Himalayan Bioresource Technology, Palampur, Himachal Pradesh 176 061, India.
authors.forename: Manju
authors.initials: M
authors.lastname: Bala
authors.affiliation: Regulatory Research Centre, Department of Biotechnology, CSIR-Institute of Himalayan Bioresource, Palampur, Himachal Pradesh, 176 061, India.
authors.forename: Kunal
authors.initials: K
authors.lastname: Pratap
authors.affiliation: Academy of Scientific and Innovative Research, New Delhi, India; Natural Product Chemistry and Process Development Division, CSIR-Institute of Himalayan Bioresource Technology, Palampur, Himachal Pradesh 176 061, India.
authors.forename: Praveen Kumar
authors.initials: PK
authors.lastname: Verma
authors.affiliation: Academy of Scientific and Innovative Research, New Delhi, India; Natural Product Chemistry and Process Development Division, CSIR-Institute of Himalayan Bioresource Technology, Palampur, Himachal Pradesh 176 061, India. Electronic address: bikram_npp@rediffmail.com.
authors.forename: Bikram
authors.initials: B
authors.lastname: Singh
authors.affiliation: Regulatory Research Centre, Department of Biotechnology, CSIR-Institute of Himalayan Bioresource, Palampur, Himachal Pradesh, 176 061, India. Electronic address: yogendra@ihbt.res.in.
authors.forename: Yogendra
authors.initials: Y
authors.lastname: Padwad
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology*; D016466:CHO Cells; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003412:Cricetulus; D006801:Humans; D007155:Immunologic Factors / Q000737:chemistry / Q000494:pharmacology*; D008519:Medicine, Traditional; D051379:Mice; D064209:Phytochemicals / Q000302:isolation & purification / Q000494:pharmacology; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology*; D018547:Plant Stems; D013154:Spleen / Q000166:cytology; D031610:Tinospora
chemical_list: D000970:Antineoplastic Agents; D007155:Immunologic Factors; D064209:Phytochemicals; D010936:Plant Extracts
keywords: Anticancer; HPTLC; Immunomodulatory; Tinospora cordifolia