This application is aimed at the development of a new class of small molecule therapeutics for human cancer. Synthetic pyrrole-imidizole polyamides have been shown to bind predetermined DNA sequences with subnanomolar affinities, comparable to the binding affinities of natural transcriptional regulatory proteins;moreover, these molecules inhibit transcription factor-DNA interactions and gene expression both in vitro and in cultured cells. A specific polyamide-DNA alkylator (chlorambucil) conjugate has been identified that alters the morphology and growth characteristics of colon carcinoma cells in culture, and causes these cells to arrest in the G2/M stage of the cell cycle, without any apparent cytotoxicity. Cells treated with this compound fail to grow in soft agar, and do not form tumors in nude mice, indicating that polyamide-treated cells are no longer tumorigenic. Importantly, this compound is active in vivo, reversing the proliferating potential of metastatic colon carcinoma cells in immunocompromised mice. Microarray analysis has given us the striking result that only one gene is significantly down regulated by this polyamide, and RT-PCR and western blotting experiments confirm that histone H4 mRNA and protein is indeed down regulated by this polyamide. To follow up on these observations, polyamide-chlorambucil conjugates will be screened for their effects on cancer cell proliferation both in cell culture and in animal models for various human cancers. The genomic targets of polyamides that inhibit cancer cell growth will be assessed using high density DNA microarray/hybridization methodology, and verified by other methods. The mechanism of inhibition of transcription by polyamides will be assessed both in vitro, in cell culture and in animal models.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA107311-05
Application #
7535177
Study Section
Bio-Organic and Natural Products Chemistry Study Section (BNP)
Program Officer
Lees, Robert G
Project Start
2004-12-03
Project End
2010-11-30
Budget Start
2008-12-01
Budget End
2010-11-30
Support Year
5
Fiscal Year
2009
Total Cost
$527,155
Indirect Cost
Name
Scripps Research Institute
Department
Type
DUNS #
781613492
City
La Jolla
State
CA
Country
United States
Zip Code
92037
Jespersen, Christine; Soragni, Elisabetta; James Chou, C et al. (2012) Chromatin structure determines accessibility of a hairpin polyamide-chlorambucil conjugate at histone H4 genes in pancreatic cancer cells. Bioorg Med Chem Lett 22:4068-71
Chou, C James; O'Hare, Thomas; Lefebvre, Sophie et al. (2008) Growth arrest of BCR-ABL positive cells with a sequence-specific polyamide-chlorambucil conjugate. PLoS One 3:e3593
Chou, C James; Farkas, Michelle E; Tsai, Sherry M et al. (2008) Small molecules targeting histone H4 as potential therapeutics for chronic myelogenous leukemia. Mol Cancer Ther 7:769-78
Tsai, Sherry M; Farkas, Michelle E; Chou, C James et al. (2007) Unanticipated differences between alpha- and gamma-diaminobutyric acid-linked hairpin polyamide-alkylator conjugates. Nucleic Acids Res 35:307-16
Alvarez, David; Chou, C James; Latella, Lucia et al. (2006) A two-hit mechanism for pre-mitotic arrest of cancer cell proliferation by a polyamide-alkylator conjugate. Cell Cycle 5:1537-48