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Target Cell Activation of a Structurally Novel NOD-Like Receptor Pyrin Domain-Containing Protein 3 Inhibitor NT-0796
Pamela Smolak1, MyTrang Nguyen1, Christine Diamond1
1Department of Genetics, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina (B.H.K., M.N., J.N.S.); Nodthera, Seattle Washington (P.S., C.D., H.W., J.R.D., K.S., C.A.G.); Nodthera, Cambridge, United Kingdom (D.H., A.P.W.); and Nodthera, Boston, Massachusetts (M.G.B.).
A novel ester-containing drug, NT-0796, effectively inhibits NLRP3 inflammasome activation in human cells by converting to its active form, NDT-19795. A humanized mouse model demonstrated NT-0796
Area of Science:
- Immunology
- Pharmacology
Background:
- The NLRP3 inflammasome is a key regulator of innate immunity and a therapeutic target for chronic inflammatory diseases.
- Endogenous NLRP3 activators are implicated in numerous human inflammatory conditions.
Purpose of the Study:
- To identify and characterize novel inhibitors of NLRP3 inflammasome activation.
- To investigate the role of carboxylesterase-1 (CES-1) in the activation and efficacy of ester-containing NLRP3 inhibitors.
- To develop a humanized mouse model for studying the pharmacokinetics and pharmacodynamics of NT-0796.
Main Methods:
- Identification of NDT-19795 as a carboxylic acid-containing NLRP3 inhibitor.
- Synthesis of isopropyl-ester prodrug NT-0796 and evaluation of its potency in human and mouse monocytes/macrophages.
- Generation of a CES-1 humanized mouse model by replacing the mouse Ces1c gene with human CES-1.
- In vitro and in vivo assessment of NT-0796 efficacy in wild-type and humanized mice.
Main Results:
- NDT-19795 inhibits NLRP3 activation in human and mouse cells.
- The ester prodrug NT-0796 exhibits enhanced potency in human cells due to CES-1 mediated conversion to NDT-19795.
- Mouse macrophages lack CES-1, rendering NT-0796 ineffective in these cells.
- Human CES-1 expression in monocytes/macrophages of engineered mice restored NT-0796 efficacy both in vitro and in vivo.
Conclusions:
- NDT-19795 is a novel NLRP3 inhibitor, and its ester prodrug NT-0796 offers enhanced cell penetration and targeted delivery.
- The development of CES-1 humanized mice provides a valuable preclinical model for evaluating ester-based therapeutics.
- The co-localization of CES-1 and NLRP3 in monocytes/macrophages allows for targeted inhibition of inflammation via ester prodrugs.
