STUDIES ON ANTIVIRAL ACTIVITY AND CYTOTOXICITY OF MORINDA CITRIFOLIA NONI

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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.
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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.
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The fruit juice is in high demand in alternative
medicine for different kinds of illnesses
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The Polynesians utilized the whole Noni plant in
various combinations for herbal remedies.
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Herbal and natural products of folk medicine have
been used for centuries in every culture
throughout the world.
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Morinda citrifolia L (Noni) is one of the
traditional folk medicinal plants that has been
used for broad range of therapeutic and
nutritionalvalue.
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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.
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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,
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• 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.
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• 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-Quinazo
  lin-4-ones.

Indian J. Heterocyclic chem. Vol.14, Jan-Mar,
2005, 255-256.
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DETAILS OF ANTIVIRAL ASSAY

• ANTIHCV ACTIVITY IN HUH 5.2 CELLS
• ANTIHIV ACTIVITY IN MT-4 CELLS

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MTT ASSAY PRINCIPLE
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96 MICROTITER PLATE ASSAY
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VIRUSES, DISEASES AND CELL LINE
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Anti HCV activity of compound on HCV Subgenomic
replicon assay in human hepatoblastoma cells
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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.
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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 culturetreated white 96-well view plate
in complete DMEM supplemented with 250 _g/mL
G418. After incubation for 24 hours at 37C (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 37C, 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 the
concentration 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.
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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 37C, after which
the cell number was determined by means of
the(3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymetho
xy phenyl)-2-(4-sulfophenyl)-2H-tetrazolium)
/phenazine methosulfate method (Promega). The
50 cytostatic concentration(CC50) was de.ned as
the concentration that inhibited the
proliferation of exponentially growing cells by
50.  
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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.
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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.
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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  
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AntiHCV Activity of Methanolic extract of
wrightia tinctoria on HCV sub genomic replicon
replication in huh-5-2-cells

• Percentage of untreated control
• Interferon a-2b at 10,000 units/well reduced the
  signal in the
• viral RNA (Luciferase) assay to background level
  without any
• cytostatic activity

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Anti HIV activity and cytotoxicity of the
compounds in MT-4 cells.
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Anti-HIV Activity and Cytotoxicity of Morinda
citrifolia  
  EC50and CC50 value are expressed in ?g/ml  
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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 activity against (MT-4) cells-Adult T
Cell leukemia with (CC50 0.1930 ?g/ml).
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Anti-HIV Activity and Cytotoxicity Of Wrightia
tinctoria
EC50, EC90 and CC50 value are expressed in ?g/ml
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FUTURE PLAN

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

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ACKNOWLEDGEMENT Dr. MYRIAM.W MOLECULAR
MEDICINE BELGIUM Dr.JOHAN NEYTS REGA INSTITUTE
FOR MEDICAL RESEARCH BELGIUM
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THANK YOUBY P.SELVAM
Asst.Prof
Dept.of Pharm.Chemistry
A.K. College of Pharmacy
Anand nagar.