Mutations in the ras family of genes were first identified in human cancer over 30 years ago. Such mutations result in the constitutive activation of one or more of three major Ras protein isoforms, including H-Ras, N-Ras, or K-Ras, that mediate important signaling pathways leading to uncontrolled cell growth and tumor development. Activating ras gene mutations occur de novo in approximately one third of all human cancers and are especially prevalent in colorectal, lung, and pancreatic tumors. Mutations in ras also arise in tumors that become resistant to chemotherapy and/or radiation. Currently there are no available drugs approved by the U.S. Food and Drug Administration that can selectively suppress the growth of tumors driven by activated Ras. From a phenotypic assay involving high-throughput screening, ADT Pharmaceuticals Inc. has discovered a novel compound series that potently and selectively inhibit tumor cells harboring activated Ras. Following extensive chemical optimization, a preclinical development candidate, DC070-547, was identified that shows low nanomolar growth inhibitory IC50 values in tumor cells having activated Ras while tumor cells lacking activated Ras, and cells derived from normal tissues, are essentially insensitive. The mechanism appears to involve direct binding within the catalytic domain of Ras to disrupt Ras-Raf interactions. Although DC070-547 shows evidence of in vivo antitumor activity in mouse xenograft models, the compound is rapidly metabolized by glucuronidation at a phenolic hydroxyl site within the molecule. We therefore propose a prodrug approach to improve the metabolic stability of DC070-547. Aim 1 will synthesize and characterize the metabolic stability of 12 unique prodrug forms of DC070-547. Aim 2 will evaluate the pharmacokinetics of candidate prodrugs in mice and will measure the levels of unchanged prodrug, as well as any active or inactive metabolites in plasma and tissues. Aim 3 will determine in vivo tolerance and antitumor activity of candidate prodrugs in xenograft models. We anticipate a clinical development candidate will emerge from this project that will be advanced to a phase II SBIR application involving GMP scale-up synthesis and GLP toxicity testing to support an IND application for human clinical trials in patients with Ras-driven cancers.
Public Health Relevance Statement: Public Health Relevance: A high percentage of human cancers harbor activating mutations in the ras family of oncogenes that lead to the formation of abnormal Ras proteins giving rise to highly aggressive and drug-resistant tumors for which there are no drugs approved by the FDA to treat Ras-driven cancers. ADT Pharmaceuticals Inc. has discovered a novel class of Ras inhibitors and identified a preclinical drug development candidate, DC070-547, that potently and selectively inhibits the growth of tumor cells with activated Ras. While DC070-547 has attractive drug-like properties and promising antitumor activity in mouse models of cancer, further chemical optimization is necessary to improve the metabolic stability of DC070-547 using a prodrug approach, which is expected to result in a highly effective and safe drug development candidate for clinical trials involving patients with Ras- driven cancers such as certain colorecta, pancreatic, and lung cancers.
Project Terms: Affinity; aqueous; base; Beta-glucuronidase; Binding; Biological Assay; Buffers; Catalytic Domain; Cells; Chemicals; chemotherapy; Clinic; Clinical; Clinical Trials; Colon Carcinoma; Colonic Neoplasms; Colorectal; Colorectal Cancer; Colorectal Neoplasms; design; Development; Dose; drug development; Drug Kinetics; Drug or chemical Tissue Distribution; Drug resistance; Ensure; Enzymes; Evaluation; Family; FDA approved; Formulation; Gene Family; Gene Mutation; Generations; Goals; Growth; Guanosine Triphosphate; HCT116 Cells; high throughput screening; HRAS gene; Human; Hydroxyl Radical; Image; improved; In Vitro; in vivo; KRAS2 gene; Lead; Life; Liver; Liver Microsomes; Luciferases; Lung Neoplasms; Malignant - descriptor; Malignant neoplasm of lung; Malignant neoplasm of pancreas; Malignant Neoplasms; Maximum Tolerated Dose; Measures; Mediating; Medical; Metabolic; Metabolism; Methods; Monitor; mouse model; Mus; mutant; Mutation; nanomolar; National Cancer Institute; Neoplasm Metastasis; neoplastic cell; Normal tissue morphology; novel; novel therapeutic intervention; Oncogenes; Oncogenic; oncology; Oral; Pancreas; pancreatic neoplasm; Parents; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Phase I Clinical Trials; Plasma; pre-clinical; Preclinical Drug Development; Primary Neoplasm; Prodrugs; Property; Protein Isoforms; public health relevance; Radiation; ras Genes; Ras Inhibitor; ras Proteins; Ras/Raf; Research; Resistance; Route; scale up; screening; Series; Signal Pathway; Signal Transduction; Site; Small Business Innovation Research Grant; small molecule; small molecule inhibitor; Solubility; subcutaneous; success; Surface; Therapeutic; Tissues; Toxic effect; Toxicity Tests; treatment effect; tumor; Tumor Cell Line; tumor growth; tumorigenesis; uncontrolled cell growth; United States Food and Drug Administration; Variant; Weight Gain; Xenograft Model; Xenograft procedure
