A Potent Fluorescent Reversible-Covalent Inhibitor of Cardiac Muscle Contraction

Fangze Cai; Thomas Kampourakis; Brittney A. Klein; Brian D. Sykes

doi:10.1021/acsmedchemlett.1c00366

A Potent Fluorescent Reversible-Covalent Inhibitor of Cardiac Muscle Contraction

Fangze Cai, Thomas Kampourakis, Brittney A. Klein, Brian D. Sykes

Internal Medicine

Research output: Contribution to journal › Article › peer-review

1 Scopus citations

Abstract

Compounds that directly modulate the response of the cardiac sarcomere have potential in the treatment of cardiac disease. While a number of sarcomere activators have been discovered and extensively studied, very few inhibitors have been identified. We report a potent cardiac sarcomere inhibitor, DN-F01, targeting the cardiac muscle thin filament protein troponin complex. Functional studies show that DN-F01 has a strong inhibitory calcium-dependent effect on cardiac myofibrillar ATPase activity with an IC50 value of 11 ± 4 nmol/L. DN-F01 is shown to bind to a cardiac troponin C-troponin I chimera (cChimera) with a KD of μ50 nM using fluorescence spectroscopy, indicating that troponin is the likely target for DN-F01. NMR titrations of DN-F01 to C35S and A-Cys cChimera show covalent and noncovalent binding of DN-F01 bound to the calcium-saturated cChimera.

Original language	English
Pages (from-to)	1503-1507
Number of pages	5
Journal	ACS Medicinal Chemistry Letters
Volume	12
Issue number	9
DOIs	https://doi.org/10.1021/acsmedchemlett.1c00366
State	Published - Sep 9 2021

Bibliographical note

Publisher Copyright:
© 2021 American Chemical Society.

Funding

Financial support from University of Alberta Faculty of Medicine Transitional Program (B.D.S.), Heart and Stroke foundation of Canada (G-14-0005884, B.D.S.), and British Heart Foundation (Fellowship FS/16/3/31887, T.K.), and University of Alberta Faculty of Medicine Motyl Graduate Studentship in Cardiac Science (F.C.). The authors thank Jack Moore for mass spectrometry experiments; Bridgette Hartley and Olivier Julien for their assistance with fluorescence experiments.

Funders	Funder number
University of Alberta Faculty of Medicine Motyl Graduate Studentship in Cardiac Science
University of Alberta
Heart and Stroke Foundation of Canada	G-14-0005884
British Heart Foundation	FS/16/3/31887

Keywords

cardiac
inhibitor
sarcomere
troponin

ASJC Scopus subject areas

Biochemistry
Drug Discovery
Organic Chemistry

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10.1021/acsmedchemlett.1c00366

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title = "A Potent Fluorescent Reversible-Covalent Inhibitor of Cardiac Muscle Contraction",

abstract = "Compounds that directly modulate the response of the cardiac sarcomere have potential in the treatment of cardiac disease. While a number of sarcomere activators have been discovered and extensively studied, very few inhibitors have been identified. We report a potent cardiac sarcomere inhibitor, DN-F01, targeting the cardiac muscle thin filament protein troponin complex. Functional studies show that DN-F01 has a strong inhibitory calcium-dependent effect on cardiac myofibrillar ATPase activity with an IC50 value of 11 ± 4 nmol/L. DN-F01 is shown to bind to a cardiac troponin C-troponin I chimera (cChimera) with a KD of μ50 nM using fluorescence spectroscopy, indicating that troponin is the likely target for DN-F01. NMR titrations of DN-F01 to C35S and A-Cys cChimera show covalent and noncovalent binding of DN-F01 bound to the calcium-saturated cChimera.",

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Y1 - 2021/9/9

N2 - Compounds that directly modulate the response of the cardiac sarcomere have potential in the treatment of cardiac disease. While a number of sarcomere activators have been discovered and extensively studied, very few inhibitors have been identified. We report a potent cardiac sarcomere inhibitor, DN-F01, targeting the cardiac muscle thin filament protein troponin complex. Functional studies show that DN-F01 has a strong inhibitory calcium-dependent effect on cardiac myofibrillar ATPase activity with an IC50 value of 11 ± 4 nmol/L. DN-F01 is shown to bind to a cardiac troponin C-troponin I chimera (cChimera) with a KD of μ50 nM using fluorescence spectroscopy, indicating that troponin is the likely target for DN-F01. NMR titrations of DN-F01 to C35S and A-Cys cChimera show covalent and noncovalent binding of DN-F01 bound to the calcium-saturated cChimera.

AB - Compounds that directly modulate the response of the cardiac sarcomere have potential in the treatment of cardiac disease. While a number of sarcomere activators have been discovered and extensively studied, very few inhibitors have been identified. We report a potent cardiac sarcomere inhibitor, DN-F01, targeting the cardiac muscle thin filament protein troponin complex. Functional studies show that DN-F01 has a strong inhibitory calcium-dependent effect on cardiac myofibrillar ATPase activity with an IC50 value of 11 ± 4 nmol/L. DN-F01 is shown to bind to a cardiac troponin C-troponin I chimera (cChimera) with a KD of μ50 nM using fluorescence spectroscopy, indicating that troponin is the likely target for DN-F01. NMR titrations of DN-F01 to C35S and A-Cys cChimera show covalent and noncovalent binding of DN-F01 bound to the calcium-saturated cChimera.

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A Potent Fluorescent Reversible-Covalent Inhibitor of Cardiac Muscle Contraction

Abstract

Bibliographical note

Funding

Keywords

ASJC Scopus subject areas

Access to Document

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