DEVELOPMENT OF A NOVEL TARGET FOR ANTIMICROBIAL DRUG

DESCRIPTION: Emergence of antibiotic-resistant microbial pathogens has created a medical crisis. Life-threatening bacterial pathogens, such as Staphylococcus aureus and Entercoccus faecalis, have developed widespread resistance to current antibiotics. Development of new antibiotics has been traditionally based on derivatizing known classes of compounds which target a limited number of cellular processes. Innovative technologies are needed to identify and develop novel antimicrobial agents for controlling antibiotic-resistant pathogens. In this Phase I proposal, the investigators propose to evaluate glutamyl-tRNA-gln amidotransferase, as a target for screening new lead compounds as potential antimicrobial agents. In several Gram-positive bacteria, this enzyme catalizes the formation of correctly aminoacylated tRNA-Gln by transfer of an amide group to the gamma-carboxylate of glutamyl-tRNA-Gln. In eukaryotic cells, the formation of glutaminyl-tRNA-Gln is catalyzed by glutamine-tRNA synthetase, providing the opportunity to develop chemotherapeutic agents specifically targeted at bacteria. $ = TOTAL AWARD AMTS & NOT LIMITED TO PORTION OF PROJECT RELATED TO SUBJECT OF SEARCH SUBPROJECT $ = TOTAL AWARD AMOUNT DIVIDED BY NUMBER OF SUBPROJECTS SOURCE: CRISP FORMAT F FY 97 LAST UPDATE 04-07-98 1QUERY 1536 ID SEARCH 06/01/98 PAGE 87 --PROJECT NUMBER......1 R43 AI41831-01 INVESTIGATOR NAME/ADDRESS FY 97 GROSSMAN, ABRAHAM IRG/INTRAMURAL UNIT..ZRG5 INVITRO DIAGNOSTICS, INC. AWARD AMOUNT......... $100,000 666 WASHINGTON AVENUE PLEASANTVILLE, NY 10570 PERFORMING ORGANIZATION: INVITRO DIAGNOSTICS, INC. TITLE TEST FOR HIV 1 REV PROTEIN USING Q-BETA SOLID TECHNOLOGY ABSTRACT: DESCRIPTION: (Adapted from Applicant's Abstract) The applicant proposes to develop a clinical analytical method for HIV-1 Rev protein. The technology is based on the combination of two relatively new technologies. First is the SELEX technology which has so far identified strong binding nucleotide sequences to Rev protein. The second is the specific amplification of Q- beta replicase of RNA containing recognized sequences by this enzyme. The idea is to make two RNA apttamers that will bind to Rev in such a way that the ends are juxtapositioned for the ligation by T4 RNA ligase. Once this is done, the ligated RNA can be amplified specifically by Q-beta replicase. The amplified product would be used as quantitation of Rev protein. For the development of such a system, the applicant proposes to construct two recombinant RNA aptamers for Rev protein binding and for the template of Q-beta replicase. Then to find conditions the aptamers anneal maximally to Rev before they anneal to themselves. This will be followed by testing in ligation and amplification. If this is successful, the methods in test-kits will be tested using clinical samples. $ = TOTAL AWARD AMTS & NOT LIMITED TO PORTION OF PROJECT RELATED TO SUBJECT OF SEARCH SUBPROJECT $ = TOTAL AWARD AMOUNT DIVIDED BY NUMBER OF SUBPROJECTS SOURCE: CRISP FORMAT F FY 97 LAST UPDATE 04-07-98 1QUERY 1536 ID SEARCH 06/01/98 PAGE 88 --PROJECT NUMBER......1 R43 AI41832-01 INVESTIGATOR NAME/ADDRESS FY 97 NEELY, CONSTANCE F IRG/INTRAMURAL UNIT..ZRG5 LINK TECHNOLOGY INC AWARD AMOUNT......... $100,000 PO BOX 97183 RALEIGH, NC 27624 PERFORMING ORGANIZATION: LINK TECHNOLOGY, INC. TITLE NOVEL ASSAY FOR MEASUREMENT OF ENDOTOXIN ABSTRACT: Endotoxin is a toxin released from certain bacteria, gram negative organisms. It may be present as a contaminant in medical devices or pharmaceutical solutions. If present in the blood of patients with septicemia, it may cause irreversible organ damage, shock, and death. Septicemia is the 13th leading cause of death in humans and accounts for 100,000 deaths annually in the United States. The only assay currently available to measure endotoxin in industrial and biological samples, Limulus amebocyte lysate (LAL) assay, is based on the reactivity of endotoxin with an amebocyte lysate isolated from the hemolymph of horseshoe crabs, Limulus polyphemus. However it is FDA approved for industrial use only. Although it is sensitive, this assay lacks specificity and is not reliable for the detection of endotoxin in biological samples, including the plasma of patients with septicemia. Using a number of highly selective radioligands for A1 adenosine receptors and membranes expressing A1 adenosine receptors, it was discovered that endotoxin displaces radioligand binding to A1 adenosine receptors. This simple radioligand binding assay is as sensitive and more specific than the LAL assay for the detection f endotoxin in saline. Currently the industrial market for the LAL assay is estimated at $60 million annually. Therefore, with this SBIR grant, the primary aim is to develop further the radioactive ligand binding assay (LBA), develop a nonradioactive LBA, and the validation of them with the LAL to measure endotoxin in pharmaceutical and industrial solutions, as well as biological products. Successful completion of the proposed experiments in this application would support a 510 (k) application for the industrial use of the LBA and lead to a Phase II SBIR grant application to support the submission of a Premarket Approval (PMA) for the clinical use of this LBA for the measurement of endotoxin in the plasma of patients with gram negative septicemia.