ANTIFUNGAL ANALOG LIBRARY

The long term goal of this project is to generate novel compounds for the treatment of fungal infections, particularly those in immuno- compromised patients. A library of analogs will be prepared based on the broad spectrum antifungal agents candicidin and FR-008. The library will constructed by manipulation of the gene cluster encoding the biosynthetic enzymes for FR-008. Selected genes encoding individual and groups of enzymatic activities from diverse polyketide synthase (PKS gene cluster scaffold. The resulting engineered PKS gene clusters will be expressed in a specially constructed streptomyces host/vector system. The analog library generated by this novel "Combinatorial Biosynthesis" technology will provide an attractive alternative to the search for new antifungal natural products, and should prove extremely valuable as a source of drug leads. Phase I of this project will demonstrate the feasibility of building an analog library based on candicidin/R-008. The first three modules of the FR-008 PKS gene cluster and genes encoding starter unit synthesis and activation will be functionally expressed in the heterologous streptomyces host. $ = 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 56 --PROJECT NUMBER......1 R43 AI40532-01A1 INVESTIGATOR NAME/ADDRESS FY 97 ZHAO, XI IRG/INTRAMURAL UNIT..ZRG5 INCELL, INC AWARD AMOUNT......... $100,000 2310 WALSH AVE SANTA CLARA, CA 95051 PERFORMING ORGANIZATION: INCELL, INC. TITLE TRANSFER ANTI-HIV RIBOZYME GENES INTO CD34+ CELLS BY EPD ABSTRACT: Gene delivery into hematopoietic stem cells is a key component of intracellular immunization as a therapeutic strategy for AIDS. Since most hematopoietic stem cells are non-dividing, efficient methods for introduction of foreign DNA into such cells are required. The proposed research is directed to the development of electronic pulse delivery technology for the effective transfer of anti-HIV ribozyme genes into human CD34+ cells. The EPD application protocols, pulse parameters and gene transfer buffers will be developed for the above purpose. $ = 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 57 --PROJECT NUMBER......1 R43 AI40742-01 INVESTIGATOR NAME/ADDRESS FY 97 PLEIMAN, CHRISTOPHER M IRG/INTRAMURAL UNIT..ZRG3 CADUS PHARMACEUTICAL CORP AWARD AMOUNT......... $92,323 1610 PIERCE ST SUITE 110 LAKEWOOD, CO 81214 PERFORMING ORGANIZATION: CADUS PHARMACEUTICAL CORPORATION TITLE DRUG SCREEN(S) TARGETING SH3 INTERACTIONS IN ALLERGY ABSTRACT: DESCRIPTION (Adapted from applicant's abstract): Allergic diseases account for more than 500,000 hospitalizations and 5,000 deaths per year in the U.S. with allergic rhinitis affecting another 20 million individuals. Allergic patients produce enhanced levels of IgE, which binds to high affinity receptors (FceRI) expressed on basophils and mast cells. Allergen exposure results in the aggregation of receptor-bound IgE triggering release of mediators that induce allergic responses. Chemotactic factors, especially IL-5, are also produced along with factors that enhance vascular permeability resulting in the late phase allergic response. Therapeutics that block the effects of FceRI and IL-5 receptor signaling will have a profound effect on these disease processes. The goal of Phase I is to design high-throughput screening (HTS) assays to discover pharmacological agents that target intracellular signal transduction events known to be essential for FceRI and IL-5 receptor function. The investigators plan to generate a recombinant GST- fusion protein containing the proline-rich region of the p85 subunit of PI-3 kinase fused to a b-galactosidase reporter. This fusion protein will be used to establish a functional binding assay with the Lyn SH3 domain in a 96- well format in preparation for Phase II HTS of diverse synthetic, combinatorial and natural product libraries to identify specific antagonists of this interaction. $ = 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 58 --PROJECT NUMBER......1 R43 AI40744-01A1 INVESTIGATOR NAME/ADDRESS FY 97 SOLER-FERRAN, DULCE IRG/INTRAMURAL UNIT..ZRG3 LEUKOSITE INC AWARD AMOUNT......... $99,999 215 FIRST STREET CAMBRIDGE, MA 02142 PERFORMING ORGANIZATION: LEUKOSITE, INC. TITLE DESIGN OF NOVEL ASTHMA THERAPEUTICS ABSTRACT: Asthma is an allergic inflammatory disorder characterized predominantly by an eosinophilic infiltrate into the airways. Eosinophils are thought to be the major effector cells mediating tissue injury and hyperresponsiveness. Therefore, it is likely that blocking the mechanisms responsible for the selective emigration of eosinophils could result in attenuation of the disease. Eotaxin, a potent and selective chemokine for eosinophils, signals through the chemokine receptor CKR3 which is exclusively expressed on eosinophils. Because of their eosinophilic specificity, eotaxin and CKR3 may play a pivotal role in the preferential migration of eosinophils into the airways. Consequently, CKR3 would appear to be an attractive target for drug development. The goal of Phase I research is to identify potent eotaxin analog antagonists. In addition, we expect to obtain information on the molecular basis for eotaxin binding and signalling through CKR3 which will be valuable for drug design. We will study the nature of the molecular recognition between eotaxin and its receptor using a mutational and functional approach. Specifically, we will: 1) Express, purify and characterize a) site directed point mutations of eotaxin, b) N-terminal extensions of eotaxin, and c) hybrid chemokines; 2) Characterize the binding and functional properties of the eotaxin mutants in in vitro assays using human eosinophils and CKR3 cell transfectants; 3) Evaluate the binding and functional properties of antagonists using eosinophils from species appropriate for in vivo asthma studies.