New Enzyme Prodrug and Methionine-Depletion Combination Therapy of Breast Cancer Designed for Effective Delivery to the Tumor
Abstract
The L-methioninase- annexin V fusion protein (FP) was produced from recombinant E. coli in good purity and relatively good yield, using the FP gene with a completely correct sequence. As indicated by measuring the dissociation constant (Kd), purified FP binds strongly to human endothelial cells, MCF-7 breast cancer cells, and MDA-MB-231 breast cancer cells grown in vitro. Tests with the enzyme prodrug over a period of 3 days showed significant cell killing at 500 microns SeMet for endothelial cells, 50 microns SeMet for MCF-7 breast cancer cells, and 10 microns SeMet for MDA-MB-231 breast cancer cells; with no FP present, significant cell killing was not observed for the endothelial cells and MDA-MB-231 cancer cells at up to 1000 microns SeMet and for MCF-7 cancer cells at up to 500 microns SeMet. These results provide strong support for the idea that this enzyme/prodrug system will lead to killing of breast tumors. Based on time profiles of the FP in the bloodstream of nude mice and on tests of injecting SeMet i.p at various levels, the FP and SeMet will both be injected at a level of 10 mg/kg for the enzyme prodrug tests in nude mice, which are currently in progress.
Document Details
- Document Type
- Technical Report
- Publication Date
- Oct 01, 2010
- Accession Number
- ADA535378
Entities
People
- Roger G. Harrison
Organizations
- University of Oklahoma