Date of Award

2019

Document Type

Thesis

Degree Name

Master of Science (M.S.)

Department

Pharmaceutical and Chemical Sciences

First Advisor

Mamoun M. Alhamadsheh

First Committee Member

Miki Susanto Park

Second Committee Member

Qinliang Zhao

Abstract

Transthyretin (TTR) amyloid cardiomyopathy (ATTR-CM) is a fatal disease with no available disease-modifying therapies. While destabilizing TTR mutations increase the risk of developing ATTR-CM, the naturally occurring Thr119Met (T119M) variant stabilizes TTR and prevent disease. The two Serine 117 and Serine 117’ (S117/S117’) side chain hydroxyl groups of monomers A and B in T119M variant TTR form direct hydrogen bonds with each other. AG10 is an orally available, small molecule TTR stabilizer in Phase 3 clinical development for ATTR-CM. Structural features that infer high potency of AG10 for stabilizing TTR remains incompletely characterized.

In order to investigate the contribution of hydrogen bonds between the pyrazole ring of AG10 and the two S117/S117’ and the salt bridges between carboxyl group and two Lysine 15 and Lysine 15’ (K15/K15’) on the stabilization of TTR, we synthesized three AG10 analogues (compounds 1, 2, and 3). Isothermal titration calorimetry (ITC) was used to determine the binding affinities (Kd) of ligands to TTR. Biochemical studies were used to compare the potency of AG10 relative to other ligands in stabilizing TTR in buffer and human serum.

Results highlight the crucial roles played by the carboxyl group and pyrazole ring of AG10 and the importance of the hydrogen bonds it forms with the two TTR dimers, mimicking the interactions in the protective T119M-TTR mutation and enhancing the kinetic stability of the TTR tetramer.

Pages

57

Available for download on Wednesday, May 12, 2021

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