Abstract 1963: Targeting hematological malignancies with two functionally and mechanistically distinct classes of cereblon mediated protein homeostatic modulators

BioTheryX's novel and unique Protein Homeostatic Modulators (PHMsTM) are small molecule “molecular glues” that promote ubiquitination of substrate proteins by the CRL4CRBN (CUL4-DDB1-RBX1-CRBN) ubiquitin ligase, leading to their subsequent degradation by the 26S proteasome. The therapeutic benefit of targeted protein degradation via cereblon (CRBN) has been clinically validated by the immunomodulatory drugs lenalidomide and pomalidomide in 5q-deletion-associated myelodysplastic syndrome and multiple myeloma, respectively. We present PHM® therapeutic candidates with novel targeted degradation profiles of disease-relevant proteins including GSPT1, Ikaros, and CK1α via CRBN with significant clinical potential in the treatment of hematological malignancies as well as solid tumors. Phenotypic screens of our proprietary PHM® library has led to the discovery of compounds that exhibit significant cytotoxicity in human AML and lymphoma cell lines, unlike classical IMiDs such as lenalidomide and pomalidomide. Cell viability assays revealed that these PHMsTM have IC50s in the low-nanomolar range. Importantly, they exhibit a large in vitro safety window with higher IC50s in normal human liver epithelial cells. The compounds are highly potent and can achieve significant AML and lymphoma selective cytotoxicity with short-term exposure of only 8 hours suggesting a highly rapid mechanism of action. Immunoblot analysis shows that the PHMsTM degrade CRBN substrates such as GSPT1, Ikaros and CK-1α and concomitantly activate the apoptosis machinery within 6 hours of treatment. The substrate degradation profile of PHMsTM combined with the cytokine profile divides the PHMsTM into two distinct mechanistic and functional classes of molecules - (1) purely cytotoxic, and (2) cytotoxic and immune-modulatory PHMsTM. Human PBMCs activated with lipopolysaccharide or α-CD3 followed by DMSO or compound treatment for 24 hours revealed that PHMsTM belonging to the latter class inhibit proinflammatory cytokines such as IL-1β, IL-6 and TNF-α as well as induce IL-2, an indicator of T cell activation, and are a 100-fold more potent than pomalidomide. In vivo efficacy of BTX-PHMsTM was evaluated using the MV-4-11 human AML xenograft model in athymic nude mice. The study shows a significant reduction in tumor volume with daily dosing, further establishing the PHMsTM as clinical candidates for AML. Of significance, is the effect of BTX1188 that completely obliterates the tumors within 30 days of