Related Experiment Videos
Cell-based screening approach for antitumor drug leads which exploits sensitivity differences between normal and
L T Vassilev1, S Kazmer, I M Marks
1Discovery Oncology, Roche Research Center, Hoffmann-La Roche Inc., Nutley, NJ 07110, USA. lyubomir.vassilev@roche.com
Abstract:
A cell-based in vitro screening approach for identification of antitumor drug leads that exploits the differential sensitivity between normal and cancer cells was developed. It is a three-step, high-throughput screen for antiproliferative and/or cytotoxic activity measured by a 7 day MTT [3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl tetrazolium bromidel assay using small panels of proliferating primary human cells and established cancer cell lines. Proof-of-concept experiments successfully identified 11 known cancer drugs randomly mixed with 5000 test compounds. Application of this screening approach to a library of 110000 compounds allowed for the identification of several novel chemical classes of compounds active against an expanded panel of cancer cell lines in vitro. Two of the compounds representing novel mitotic inhibitors with in vivo potency against established breast cancer xenografts (MDA-MB-435) are reported here.
Insights
A novel cell-based screening method identifies new antitumor drug leads by testing their effects on cancer versus normal cells. This approach successfully found known drugs and novel compounds, including two potent mitotic inhibitors for breast cancer xenografts.
Area of Science:
- Oncology
- Drug Discovery
- Cell Biology
Background:
- Developing effective antitumor drugs requires identifying compounds with selective toxicity towards cancer cells.
- High-throughput screening (HTS) methods are crucial for efficiently evaluating large compound libraries for potential drug leads.
Purpose of the Study:
- To develop and validate a cell-based, in vitro screening approach for identifying novel antitumor drug candidates.
- To exploit differential drug sensitivity between normal and cancer cells for selective drug lead identification.
Main Methods:
- A three-step, high-throughput screening (HTS) assay was designed.
- Antiproliferative and cytotoxic activities were measured over 7 days using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay.
- The assay utilized panels of proliferating primary human cells and established cancer cell lines.
Main Results:
- Proof-of-concept experiments correctly identified 11 known anticancer drugs from a mixture of 5000 compounds.
- Screening a library of 110,000 compounds identified novel chemical classes with in vitro activity against an expanded panel of cancer cell lines.
- Two novel compounds identified as mitotic inhibitors demonstrated in vivo potency against breast cancer xenografts (MDA-MB-435).
Conclusions:
- The developed cell-based screening approach is effective for identifying potential antitumor drug leads.
- The identified novel mitotic inhibitors show promise for further development as breast cancer therapeutics.