In Vitro Pharmacology

Oncology

India’s largest tumor bank repository with human cancer cell lines representing 15 – 20 different cancer types
Characterized cell lines from authenticated sources
Inclusive of untransformed cell lines to enable early safety assessment
Drug resistant cell lines & primary cultures from human biopsies
Complete repertoire of viability & cytotoxicity assessment screens
Customized Molecular target based screens
Combination index studies for assessment of synergy

In Vitro Services

Model	Evaluation of drug interactions in combination cancer chemotherapy by combination index method
Test system	Cancer cell lines
Methods	Determination of % cytotoxicity & IC50 of individual drugs Selection of concentrations around IC50 for combination studies Testing all possible combinations of drugs for combined cytotoxicities Calculation of Combination Index (CI) CI=1 – Synergy CI=1 – Additive CI=1 – Antagonism
End points	Categorization of drug combinations as synergistic/additive/antagonistic for each combination tested.

Model	DETERMINATION OF SYNERGISTICALLY EFFICACIOUS COMBINATION(S) OF CISPLATIN AND DOXORUBICIN IN HepG-2 CELL LINE IN VITRO BY COMBINATION INDEX METHOD
Test system	Hepatocellular carcinoma (HepG-2)
Methods	Cisplatin Doxorubicin
End points	Cells were treated at fixed IC50 of one drug (Cisplatin/Doxorubicin) in combination with variable concentrations of the other (Cisplatin/Doxorubicin). MTT assay CI value was calculated to assess the synergistic effect of test items Synergism, additive or antagonism was based on the following criteria: CI = 1 Synergism CI=1 Additive CI=1 Antagonism

Model	in vivo: model to quantitate intracellular uptake and localization of drugs (e.g. Nanoparticle/liposomal formulations)
Test system	Human tumor cell lines Human normal cells/cell lines
Methods	Treatment of target cell lines with drug Quantitative analysis – Visualization of Cellular uptake, delivery and subcellular distribution of drugs in target cell lines by TEM Transmission Electron Microscopy)/ Fluorescence microscopy Quantitative analysis – Measurement of intracellular concentrations of drug by bioanalytical methods (HPLC and LC/MS)
End points	Intracellular accumulation of drugs Sub cellular distribution/localization Uptake in non target tissues Dose/Time kinetic profiles for drug uptake Correlation with biological activity Uptake/Transport by endocytotic/Non endocytotic routes

Model	Assessment of anti-cancer activity of the compounds using tube formation assay
Test system	HUVEC Cells
Methods	Seeding of HUVEC cells on matrigel Application of vehicle and compound of interest at appropriate concentrations Validation with Combretastatin A4 or client defined reference standards
End points	Quantitative analysis of – Branch area Branch points Tube length Photomicrographs

Cell Migration assay

Model	Assessment of anti-cancer activity of the compounds using cell migration assay
Test system	HUVEC Cells
Method	Seeding of HUVEC cells Creation of wound on surface of culture Application of vehicle and Test item at appropriate concentrations
End points	Number of cells migrated to wound area

In vitro Angiogenesis model

Model	Assessment of anti-cancer activity of the compounds using tube formation assay
Test system	HUVEC Cells
Method	Seeding of HUVEC cells on matrigel Application of vehicle and compound of interest at appropriate concentrations Validation with Combretastatin A4 or client defined reference standards
End points	Quantitative analysis of – Branch area Branch points Tube length Photomicrographs

In vitro angiogenesis VEGF

Model	Assessment of VEGF, bFGF & Endostatin
Test system	HUVEC, MDA-MB-231, MCF-7, HCT-116 etc
Method	Seeding of predetermined number of Cell Type Application of vehicle and Test item at appropriate concentrations Incubation for 48 to 72hr Estimation of VEGF, bFGF & endostatin concentration in culture supernatants
End points	% Inhibition IC50 value

Model	Complete repertoire of Screens for assessment of Programmed cell death to enable selection of Proapoptotic & antiapoptotic molecules
Test system	Human normal cells/cell lines Human cancer cell lines
Apoptotic Pathways	Death receptor and mitochondrial pathway
End points	Externalization of Phosphatidyl Serine (PS) on plasma membrane by Annexin-V staining Loss of transmembrane mitochondrial potential by JC-1 staining Quantitation of Reactive oxygen species (ROS) Levels of pro-apoptotic and anti-apoptotic proteins Ras-ERK kinases Caspase-3 activation PARP cleavage DNA fragmentation Micronuclei staining Cell Cycle analysis

Model	Assessment of anti-leukemic activity of the compounds in SCID mice
Test system	SCID mice
Models	Intravenous inoculation of leukemic cell line Administration of test item and reference item at predetermined time Observation of animals for defined time period Observation of animals in satellite group for occurrence of chloroma
End points	Percentage Change in body weight Survival rate of animals Clinical signs & symptoms Quantification by flow cytometry analysis in bone marrow & peripheral blood — Standard surface specific antigen staining – CD45 — Lineage specific antigen staining Histopathology

Core team of Biochemists, Cell biologists & Molecular Biologists to Develop customized client defined target based screens Repertoire of screens

Target Classes