MODIFIED NUCLEOSIDES IN TRIPLEX FORMING OLIGONUCLEOTIDES

Award Information
Agency: Department of Health and Human Services
Branch: N/A
Contract: N/A
Agency Tracking Number: 19009
Amount: $50,000.00
Phase: Phase I
Program: SBIR
Awards Year: 1992
Solicitation Year: N/A
Solicitation Topic Code: N/A
Solicitation Number: N/A
Small Business Information
Triplex Pharmaceutical
9391 Grogans Mill Road, The Woodlands, TX, 77380
DUNS: N/A
HUBZone Owned: N
Woman Owned: N
Socially and Economically Disadvantaged: N
Principal Investigator
 Ganapathi R Revankar
 (713) 363-8761
Business Contact
Phone: () -
Research Institution
N/A
Abstract
THE PRIMARY GOAL OF THIS STUDY IS TO CHEMICALLY REFINE A CLASS OF SEQUENCE-SPECIFIC, DNA BINDING, TRIPLE HELIX-FORMING OLIGONUCLEOTIDES (TFOS). THE UNIQUE FEATURE OF THESE TFOS IS THAT BINDING OF THE THIRD STRAND TO ITS DUPLEX DNA TARGET IS PH INDEPENDENT AND IS STABILIZED BY GGC AND TAT TRIPLETS SUCH THAT THE PREFERRED ORIENTATION PLACES THE BOUND TFO ANTIPARALLEL WITH RESPECT TO THE MORE PURINE RICH STRAND OF THE UNDERLYING DUPLEX. IN ACCORD WITH MOLECULAR MODELING, 2'-DEOXYFORMYCI (DF) IS INTRODUCED INTO 3'-AMINE MODIFIES TFOS AT SITES ENGAGED IN H-BONDING WHERE POLYPURINE/POLYPRYIMIDINE RUNS IN A DUPLEX TARGET ARE INTERRUPTED BY A CG INVERSION. THE BINDING AFFINITY OF SUCH A DF-TFO IS ASSESSED BY BAND SHIFT ANALYSIS AND BY QUANTITATIVE DNASE I FOOT-PRINTING TECHNIQUES. THE EFFECT OF DF UPON TRIPLEX STRUCTURE IS ASSESSED BY LOCAL HIGH AFFINITY CUPHENANTHROLINE CLEVAGE. IN PRELIMINARY STUDIES, A HOMOLOGUE OF 3'A-PRE2AP DISPLAYED A 10-FOLD INCREASE IN BINDING AFFINITY DUE TO INCLUSION OF DF AT THREE SITES AS COMPARED TO ITS UNMODIFIED COUNTERPART. IN PHASE I STUDIES WE WILL CONFIRM THESE FINDINGS AND EXTEND THE USE OF DF SUBSTITUTION WITHIN TFOS WHICH ARE SPECIFIC FOR EPIDERMAL GROWTH FACTOR RECEPTOR (EGFR) (EG36AP), MOUSE INSULIN RECEPTOR (IR) PROMOTER (IR2AP) AND HIV-1 (HIV38P) DOMAINS. IF DF SUBSTITUTION IMPROVES TRIPLE HELIX STRUCTURE AND BINDING AFFINITY, IN PHASE II STUDIES THE BIOLOGICAL EFFECTS OF SUCH TFOS WILL BE ASSESSED.

* information listed above is at the time of submission.

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