Development of N tert Butyl hydroxylamine NtBuHA as a therapeutic agent for treating Infantile Neuronal Ceroid Lipofuscinosis INCL

PROJECT SUMMARY ABSTRACT There is a significant unmet medical need for a therapy to treat patients with Infantile Neuronal Ceroid Lipofuscinosis type CLN Batten Disease for which there are andlt in estimated patients in the United States that have no current disease treating options We will begin a pre clinical program on one small molecule that has been selected as a lead compound by a senior investigator at the NICHD Dr Anil Mukherjee This small molecule N tert butyl hydroxylamine NtBuHA has been selected amongst a panel of hydroxylamine derivatives for its minimal in vitro and in vivo toxicity profile its solubility in aqueous solution and its superior chemical activity in replacing deficient Palmitoyl Protein Thioesterase PPT activity that underlies the pathophysiology of INCL patients In preparation for an IND enabling program we will de risk NtBuHA by completing physiochemical analysis DMPK studies biodistribution studies and pilot toxicology studies Solubility stability and lipophilicity studies will be conducted in vitro to confirm NtBuHA has adequate chemical properties prior to further in vivo work Plasma protein binding studies from four species mouse rat dog human will determine thermodynamic binding parameters of NtBuHA to be used in PK analyses and that will help provide an estimation of human systemic exposure from animal data Metabolic stability will be performed in microsomes and hepatocytes of four species mouse rat dog human to estimate cross species to human phase I and phase II metabolism respectively Metabolite profiling in hepatocytes of four species mouse rat dog human will be performed to aid in the selection of toxicology species and to identify any human specific metabolites INCL patients are often treated with a combination of anti epileptic pain and anti spasticity medications e g valproate fentanyl baclofen tizanidine thus necessitating drug drug interaction CYP inhibition and induction studies that will be performed in human liver samples We will complete PK studies in rats to determine the dose dependent pharmacokinetic parameters that are both adsorption dependent via oral gavage route and adsorption independent via intravenous route as well as to estimate both human PK parameters and dose regimen needed for pharmacological response Dose escalating biodistribution studies in rats will be conducted with focus on accumulation of NtBuHA within the brain in order to determine the minimum dose required to achieve therapeutically relevant concentrations in brain tissues A day maximum tolerated dose MTD study will be completed to establish the therapeutic window for NtBuHA and ensure therapeutically relevant brain concentrations can be safely achieved The hERG AMES and micronucleus tests will be completed as in vitro toxicology gating studies that will aid in determining whether a phase II SBIR submission is merited Upon the conclusion the proposed work we will have a lead compound that is characterized for more comprehensive formulation safety and efficacy evaluation in vivo Upon successful completion of these studies we intend to enter into Phase II SBIR studies that will employ GLP facilities and GMP materials longer repeat dosing toxicity protocols in large animals and safety studies cardiac respiratory etc that will more precisely predict NtBuHA behavior in humans and that will support a subsequent IND application Ultimately we hope NtBuHA can become a marketed product that has clinically meaningful impact on INCL patients PROJECT NARRATIVE Fewer than in children born in the United States are diagnosed with INCL a devastating genetic disorder that is often diagnosed in infancy and is uniformly degenerative and fatal by late childhood We propose to develop the first small molecule therapeutic option for this disease which replaces lost Palmitoyl Protein Thioesterase activity that is the underlying cause of INCL This small molecule works through a hydroxylamine mediated chemical reaction that cleaves thioester bonds between palmitate and protein By replacing lost enzymatic activity we hope to eliminate the accumulation of palmitoyl protein inclusions in lysosomes slow the progression of neurodegeneration and achieve clinically meaningful benefit for patients and families afflicted with INCL