Secondary metabolites are usually produced by an organism as part of a defense mechanism or to facilitate reproduction. The chemical diversity of secondary metabolites mirrors the variety of enzymatic reactions present in the biosynthetic pathways. Therefore, these pathways are a constant and rich source of exciting novel transformations and unprecedented enzymes. In addition, the genetic and enzymatic knowledge acquired by elucidating the biosynthetic pathways can be exploited for the production of chemically varied secondary metabolites. Pyrrolo[1,4]benzodiazepines, a class of potent antitumor antibiotics found in actinomycetes, are sequence selective DNA alkylating agents. Phase 1 human clinical trials of SJG-136 for the treatment of metastatic or unresectable solid tumors were started in 2004. Microbiology, molecular biology and enzymological techniques will be used in this proposal to elucidate the biochemistry underlying pyrrolo[1,4]benzodiapines'production. Specifically, we propose to 1) identify the biosynthetic gene clusters for tomaymycin and sibiromycin, 2) produce novel glycosylated PBDs and PBD dimers, and 3) characterize the unique enzymatic transformations present in the biosynthetic pathway.

Public Health Relevance

Pyrrolobenzodiazepines (PBDs) are compounds naturally produced by bacteria with potent antitumor properties. PBDs'remarkable broad spectrum of activities and effectiveness against a wide variety of cancers encourages the development of new PBDs. For instance, SJG-136 is currently in phase I clinical trials against metastatic and unresectable solid tumors. We are proposing to study how nature carries out the synthesis of these compounds. The genetic and enzymological knowledge so obtained can then be used for production of new analogs of these compounds. Examples of such application are discussed in this proposal.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM084473-05
Application #
8228017
Study Section
Synthetic and Biological Chemistry B Study Section (SBCB)
Program Officer
Anderson, Vernon
Project Start
2008-05-01
Project End
2012-06-30
Budget Start
2012-03-01
Budget End
2012-06-30
Support Year
5
Fiscal Year
2012
Total Cost
$62,481
Indirect Cost
$20,411
Name
University of Maryland College Park
Department
Chemistry
Type
Schools of Earth Sciences/Natur
DUNS #
790934285
City
College Park
State
MD
Country
United States
Zip Code
20742
Gerratana, Barbara (2012) Biosynthesis, synthesis, and biological activities of pyrrolobenzodiazepines. Med Res Rev 32:254-93
Yonemoto, Isaac T; Li, Wei; Khullar, Ankush et al. (2012) Mutasynthesis of a potent anticancer sibiromycin analogue. ACS Chem Biol 7:973-7
Burks, Elizabeth A; Yan, Wupeng; Johnson Jr, William H et al. (2011) Kinetic, crystallographic, and mechanistic characterization of TomN: elucidation of a function for a 4-oxalocrotonate tautomerase homologue in the tomaymycin biosynthetic pathway. Biochemistry 50:7600-11
Connor, Katherine L; Colabroy, Keri L; Gerratana, Barbara (2011) A heme peroxidase with a functional role as an L-tyrosine hydroxylase in the biosynthesis of anthramycin. Biochemistry 50:8926-36
Li, Wei; Khullar, Ankush; Chou, Shenchieh et al. (2009) Biosynthesis of sibiromycin, a potent antitumor antibiotic. Appl Environ Microbiol 75:2869-78
Li, Wei; Chou, ShenChieh; Khullar, Ankush et al. (2009) Cloning and characterization of the biosynthetic gene cluster for tomaymycin, an SJG-136 monomeric analog. Appl Environ Microbiol 75:2958-63