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STUDIES ON ANTIVIRAL ACTIVITY AND CYTOTOXICITY OF MORINDA CITRIFOLIA NONI

STUDIES ON ANTIVIRAL ACTIVITY AND CYTOTOXICITY OF MORINDA CITRIFOLIA NONI P.SELVAM M.PHARM (PH.CHEM),(PH.D).,FISAR., Assistant Professor, Dept.of Pharmaceutical Chemistry A.K. College of Pharmacy Anand nagar.

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STUDIES ON ANTIVIRAL ACTIVITY AND CYTOTOXICITY OF MORINDA CITRIFOLIA NONI

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  1. STUDIES ON ANTIVIRAL ACTIVITY AND CYTOTOXICITY OF MORINDA CITRIFOLIA NONI P.SELVAM M.PHARM (PH.CHEM),(PH.D).,FISAR., Assistant Professor,Dept.of Pharmaceutical ChemistryA.K. College of PharmacyAnand nagar.

  2. Morinda citrifolia L (Noni) has been used in folk remedies by Polynesians for over 2000 years, and is reported to have a broad range of therapeutic effects.

  3. The fruit juice is in high demand in alternative medicine for different kinds of illnesses

  4. The Polynesians utilized the whole Noni plant in various combinations for herbal remedies.

  5. Herbal and natural products of folk medicine have been used for centuries in every culture throughout the world.

  6. Morinda citrifolia L (Noni) is one of the traditional folk medicinal plants that has been used for broad range of therapeutic and nutritionalvalue.

  7. Morinda citrifolia L (Noni) has been used in folk remedies by Polynesians for over 2000 years, and is reported have a broad range of therapeutic effects, including Antibacterial, Antiviral, Antifungal, Antitumor, Antihelmin, analgesic, Hypotensive, anti-inflammatory, Immune enhancing effects.

  8. Medicinal use of Noni : The fruit juice is in high demand in alternative medicine for different kinds of illnesses such as Arthritis, Diabetes, high blood pressure,muscle aches menstrual difficulties,Headaches, Heart disease, AIDS, Cancers, Gastric ulcers, Sprains, Mental depression, Poor digestion, Atherosclerosis,

  9. P. Selvam et al , Synthesized anti-HIV activity of 4-[(1,2-dihydro-2-oxo-3H-indol-3-ylidene)amino]-N(4,6-dimethyl-2-pyrimidinyl)-benzene sulfonamide and its derivatives. Euro. J. Pharm. Sci. 14 (2001) 313-316.

  10. P. Selvam et al , synthesized cytostatic activity of some 3-[5-amino-6-(2,3-dichlorophenyl)-[1,2,4] Triazin-3-yl]-6,8-Dibromo-2-substituted-3H-Quinazolin-4-ones. Indian J. Heterocyclic chem. Vol.14, Jan-Mar, 2005, 255-256.

  11. DETAILS OF ANTIVIRAL ASSAY • ANTIHCV ACTIVITY IN HUH 5.2 CELLS • ANTIHIV ACTIVITY IN MT-4 CELLS

  12. MTT ASSAY PRINCIPLE

  13. 96 MICROTITER PLATE ASSAY

  14. VIRUSES, DISEASES AND CELL LINE

  15. Anti HCV activity of compound on HCV Subgenomic replicon assay in human hepatoblastoma cells

  16. Hepatitis C virus (HCV) is an enveloped singlestranded(-) RNA virus that belongs to the separate genus Hepacivirus of the family Flaviviridae. HCV causes acute and chronic liver disease, including chronic hepatitis, cirrhosis, and hepatocellular carcinoma. Worldwide more than 170 million people are chronically infected with HCV and are thus at increased risk of developing serious life-threatening liver disease. Current standard therapy for chronic hepatitis C consists of the combination of pegylated interferon alpha (IFN-_) 2a in combination with ribavirin.

  17. Anti-HCV Assay in Huh 5-2 Cells. Huh 5-2 cells were seeded at a density of 5 x 103 per well in a tissue culture–treated white 96-well view plate in complete DMEM supplemented with 250 _g/mL G418. After incubation for 24 hours at 37°C (5% CO2) medium was removed and 3-fold serial dilutions in complete DMEM (without G418) of the test compounds were added in a total volume of 100 _L. After 4 days of incubation at 37°C, cell culture medium was removed and luciferase activity was determined using the Steady-Glo luciferase assay system the luciferase signal was measured using a Luminoskan Ascent. The 50% effective concentration (EC50) was de.ned as theconcentration of compound that reduced the luciferase signal by 50%. 50% effective concentration (EC50) was de.ned as the concentration of compound that reduced the luciferase signal by 50%.

  18. Cytostatic Assay. Huh 5-2, monocells were seeded at a density of 5 x103 cells per well of a 96-well plate in complete DMEM with the appropriate concentrations of G418 or hygromycin. Serial dilutions of the test compounds in complete DMEM without or G418 or hygromycin were added 24 hours after seeding.Cells were allowed to proliferate for 3 days at 37°C, after which the cell number was determined by means of the(3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxy phenyl)-2-(4-sulfophenyl)-2H-tetrazolium) /phenazine methosulfate method (Promega). The 50% cytostatic concentration(CC50) was de.ned as the concentration that inhibitedthe proliferation of exponentially growing cells by 50%.

  19. Cell growth* Viral RNA* Concentration 50 µg/ml 78 13 10 105 91 2.0 100 97 0.40 99 78 0.08 92 100 Results CC50 EC50 > 50 23 Activity of the compounds on HCV subgenomic replicon replication in huh-5-2 cells *(% of untreated control) Interferon alfa-2b at 10,000 units/well reduced the signal in the viral RNA (luciferase ) assay to background levels; without any cytostatic activity.

  20. invitro antiviral activity against HCV The compound reduced the viral RNA below 25% and promoted cell growth more than 85 % with respect to the untreated control, considered as positive antiviral activity.

  21. RESULTS AND DISCUSSION The 50% effective concentration for inhibition of HCV subgenomic replicon replication in Huh 5-2 cells (luciferase assay) by noni was 23 µg/mL. The concentration that reduced the growth of exponentially proliferating Huh 5-2 cells by 50% was greater than 50 µg/mL

  22. AntiHCV Activity of Methanolic extract of wrightia tinctoria on HCV sub genomic replicon replication in huh-5-2-cells • Percentage of untreated control • Interferon α-2b at 10,000 units/well reduced the signal in the • viral RNA (Luciferase) assay to background level without any • cytostatic activity

  23. In vitro antiHIV activity and cytotoxicity studies VIRUS : HIV-1 HTLV-IIIB CELL LINE: MT-4 CELLS ASSAY: MTT METHOD EC50,CC50 MAXIMUM PROTECTION

  24. Anti HIV activityand cytotoxicity of the compounds in MT-4 cells.

  25. COMPOUND STRAIN EC50 CC50 MAXIMUM PROTECTION MC IIIB >0.1920 0.1920 19 MC IIB >0.1930 0.1930 17.3 Anti-HIV Activity and Cytotoxicity of Morinda citrifolia EC50and CC50 value are expressed in g/ml

  26. RESULTS Morinda citrifolia has been evaluated for its anti-HIV activity and cytotoxicity against HIV-1(IIIB) replication in acutely infected MT-4 cells. Morinda citrifolia exhibited a maximum protection of 18 against HIV-1 (IIIB) strains in acutly infected MT-4 cells. Morinda citrifolia displayed distinct cytostatic activityagainst (MT-4) cells-Adult T Cell leukemia with (CC50 =0.1930g/ml).

  27. Anti-HIV Activity and Cytotoxicity Of Wrightia tinctoria EC50, EC90 and CC50 value are expressed in g/ml

  28. FUTURE PLAN • ANTIVIRAL ACTIVITY AGAINST SARS-CoV in VERO CELLS • ANTIVIRAL ACTIVITY AGAINST AVIAN FLU(H5N1) IN MDCK CELLS

  29. ACKNOWLEDGEMENT Dr. MYRIAM.W MOLECULAR MEDICINE BELGIUM Dr.JOHAN NEYTS REGA INSTITUTE FOR MEDICAL RESEARCH BELGIUM

  30. THANK YOU…BY P.SELVAM Asst.Prof Dept.of Pharm.Chemistry A.K. College of Pharmacy Anand nagar.

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