Abstract 7: A Subcutaneous, Potent and Durable RNAi Platform Targeting Metabolic Diseases, Genes PCSK9, ApoC3 and ANGPLT3

Aim: There remains high unmet medical need for therapies to treat cardio/metabolic diseases. We validated in human trials, a platform for reducing the synthesis of genes expressed in the liver. The platform utilizes a GalNAc ligand attached to the 3’ end of the sense strand of an RNAi molecule to enable delivery specifically to the liver. Here we extend the platform to targets of interest in cardiovascular disease, including PCSK9, ANGPLT3 and ApoC3. METHODS: Chemically modified siRNAs were designed and were screened for potency in vitro . pM active siRNA molecules were developed targeting PCSK9, ANGPLT3 and ApoC3. The siRNAs were tested in either rodents or in non-human primates (NHPs) for activity. RESULTS: In NHPs a single dose of ALN-PCSsc at 6 mg/kg reduced PCSK9 levels up to 97% and LDL-C up to 67%. Moreover the nadir effect (without any rebound of LDL-C) lasted >30 days indicating that once a month or longer dosing frequency in clinic should be supported. Multidose studies in NHP at 2mg/kg reduced circulating PCSK9 levels up to 94% with a subsequent lowering of LDL-C up to 67%. The effects on both PCSK9 and LDL-C was also very durable with levels of LDL-C returning to baseline > 140 days post the last dose. Safety studies in rat at doses up to 225mg/kg (multi-dose) indicate that ALN-PCSsc is safe demonstrating a very wide therapeutic index. ALN-PCSsc was selected as a development candidate and is being advanced towards an IND ALN-ANGsc (an siRNA targeting ANGPTL3) was tested in two models of hyperlipidemia, the OB/OB mouse and the hCETP-ApoB mouse. In the Ob/Ob model, treatment with ALN-ANGsc at 3mg/kg resulted in a significant lowering ANGPLT3 protein (>95%), total cholesterol(>60%), and triglycerides (>85%). Finally, we have developed a prototype molecule targeting Apoc3 with an ED90 for ApoC3 protein of <2.5mg/kg showing 50% lowering of triglycerides in and db/db mouse model of hypertriglyceridemia. CONCLUSION: We have developed a modular, robust and durable platform for the delivery of RNAi therapeutics to the liver. This platform is administered as a small volume subcutaneous dose and has a remarkable duration of effect in rodent NHP models. We have extended this platform to several targets of high interest includingPCSK9,ANGPLT3 and ApoC3.