The primary objective of the proposed research is to study the mechanism of action of clinically significant new anti-tumor FR900482 (1a), FK973 (1b) and FR66979 (1c) (FK973, 1b was the first derivative to go to clinical trials). FK973 has been shown to cross-link double-stranded DNA and cross-link DNA to DNA-binding proteins in L1210 cells. Efforts will be directed at elucidating in complete mechanistic detail, the precise mechanism of the in vitro reductive activation of FR900482 that results in covalent modification of DNA. In the previous funding period, we have determined the sequence specificity of DNA cross-link formation by FR900482 and the natural reduction product FR66979 and have determined that reduction of these substances leads to the production of a mitosene derivative that preferentially cross-links DNA at 5'-dC-dG3' boxes. The interaction of FK973 with DNA complexed to various DNA-binding proteins will also be examined in detail. Synthetic DNA substrates will be constructed and incubated with their respective DNA-binding proteins in the presence of FK793 (or FR900482); subsequent enzymatic digestion of the cross-linked nucleotide-drug-amino acid adduct will be examined to isolate and characterize the structure of the covalent cross-link. Synthetic methodology developed during the first funding cycle will be utilized to complete a total synthesis of FR900482 (1a) and FR66979 (2); this technology will be applied to the synthesis of an isotopically-labeled form of the drug for use in elucidating the structure of the DNA-drug- protein cross-link. A new class of """"""""latent"""""""" triggered mitosenes will be synthesized and utilized as potential new anti-cancer drugs and probes for the macromolecular cross-links.

National Institute of Health (NIH)
National Cancer Institute (NCI)
Research Project (R01)
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Medicinal Chemistry Study Section (MCHA)
Program Officer
Beisler, John A
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Colorado State University-Fort Collins
Schools of Arts and Sciences
Fort Collins
United States
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Bass, Phillip D; Gubler, Daniel A; Judd, Ted C et al. (2013) Mitomycinoid alkaloids: mechanism of action, biosynthesis, total syntheses, and synthetic approaches. Chem Rev 113:6816-63
Williams, Robert M (2011) Natural products synthesis: enabling tools to penetrate Nature's secrets of biogenesis and biomechanism. J Org Chem 76:4221-59
Williams, Robert M; Stille, J; Echavarren, A et al. (2011) Discussion Addendum for: 4-Methoxy-4'-nitrophenyl. Recent Advances in the Stille Biaryl Coupling Reaction and Applications in Complex Natural Products Synthesis. Organic Synth 88:197-201
Subramanian, Vidya; Williams, Robert M; Boger, Dale L et al. (2010) Methods to characterize the effect of DNA-modifying compounds on nucleosomal DNA. Methods Mol Biol 613:173-92
Chamberland, Stephen; Grüschow, Sabine; Sherman, David H et al. (2009) Synthesis of potential early-stage intermediates in the biosynthesis of FR900482 and mitomycin C. Org Lett 11:791-4
Gubler, Daniel A; Williams, Robert M (2009) Synthetic Studies Towards the Mitomycins: Construction of the Tetracyclic Core via a Reductive Aminocyclization Reaction. Tetrahedron Lett 50:4265-4267
Subramanian, Vidya; Ducept, Pascal; Williams, Robert M et al. (2007) Effects of photochemically activated alkylating agents of the FR900482 family on chromatin. Chem Biol 14:553-63
Namiki, Hidenori; Chamberland, Stephen; Gubler, Daniel A et al. (2007) Synthetic and biosynthetic studies on FR900482 and mitomycin C: an efficient and stereoselective hydroxymethylation of an advanced benzazocane intermediate. Org Lett 9:5341-4
Judd, Ted C; Williams, Robert M (2004) A concise total synthesis of (+)-FR900482 and (+)-FR66979. J Org Chem 69:2825-30
Williams, Robert M; Ducept, Pascal (2003) Interstrand cross-linking of DNA by FK317 and its deacetylated metabolites FR70496 and FR157471. Biochemistry 42:14696-701

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