2-Mercapto-Quinazolinones as Inhibitors of Type II NADH Dehydrogenase and Mycobacterium tuberculosis: Structure-Activity Relationships, Mechanism of Action and Absorption, Distribution, Metabolism, and Excretion Characterization

Dinakaran Murugesan, Peter C. Ray, Tracy Bayliss, Gareth A. Prosser, Justin R. Harrison, Kirsteen Green, Candice Soares De Melo, Tzu Shean Feng, Leslie J. Street, Kelly Chibale, Digby F. Warner, Valerie Mizrahi, Ola Epemolu, Paul Scullion, Lucy Ellis, Jennifer Riley, Yoko Shishikura, Liam Ferguson, Maria Osuna-Cabello, Kevin D. Read & 10 others Simon R. Green, Dirk A. Lamprecht, Adrie J.C. Steyn, Thomas R. Ioerger, Jim Sacchettini, Kyu Y. Rhee, Kriti Arora, Clifton E. Barry, Paul G. Wyatt, Helena I.M. Boshoff

Research output: Contribution to journalArticle

  • 2 Citations

Abstract

Mycobacterium tuberculosis (MTb) possesses two nonproton pumping type II NADH dehydrogenase (NDH-2) enzymes which are predicted to be jointly essential for respiratory metabolism. Furthermore, the structure of a closely related bacterial NDH-2 has been reported recently, allowing for the structure-based design of small-molecule inhibitors. Herein, we disclose MTb whole-cell structure-activity relationships (SARs) for a series of 2-mercapto-quinazolinones which target the ndh encoded NDH-2 with nanomolar potencies. The compounds were inactivated by glutathione-dependent adduct formation as well as quinazolinone oxidation in microsomes. Pharmacokinetic studies demonstrated modest bioavailability and compound exposures. Resistance to the compounds in MTb was conferred by promoter mutations in the alternative nonessential NDH-2 encoded by ndhA in MTb. Bioenergetic analyses revealed a decrease in oxygen consumption rates in response to inhibitor in cells in which membrane potential was uncoupled from ATP production, while inverted membrane vesicles showed mercapto-quinazolinone-dependent inhibition of ATP production when NADH was the electron donor to the respiratory chain. Enzyme kinetic studies further demonstrated noncompetitive inhibition, suggesting binding of this scaffold to an allosteric site. In summary, while the initial MTb SAR showed limited improvement in potency, these results, combined with structural information on the bacterial protein, will aid in the future discovery of new and improved NDH-2 inhibitors.

LanguageEnglish
Pages954-969
Number of pages16
JournalACS Infectious Diseases
Volume4
Issue number6
DOIs
Publication statusPublished - 8 Jun 2018

Fingerprint

Quinazolinones
Structure-Activity Relationship
Mycobacterium tuberculosis
Adenosine Triphosphate
Allosteric Site
Bacterial Proteins
Enzymes
Electron Transport
Microsomes
Oxygen Consumption
NAD
Membrane Potentials
Energy Metabolism
Biological Availability
Glutathione
Pharmacokinetics
NADH dehydrogenase II
Electrons
Mutation
Membranes

Keywords

  • mercapto-quinazolinones
  • Mycobacterium tuberculosis
  • respiration
  • small molecule NDH-2 inhibitors
  • structure-activity relationship
  • type II NADH dehydrogenase

Cite this

Murugesan, Dinakaran ; Ray, Peter C. ; Bayliss, Tracy ; Prosser, Gareth A. ; Harrison, Justin R. ; Green, Kirsteen ; Soares De Melo, Candice ; Feng, Tzu Shean ; Street, Leslie J. ; Chibale, Kelly ; Warner, Digby F. ; Mizrahi, Valerie ; Epemolu, Ola ; Scullion, Paul ; Ellis, Lucy ; Riley, Jennifer ; Shishikura, Yoko ; Ferguson, Liam ; Osuna-Cabello, Maria ; Read, Kevin D. ; Green, Simon R. ; Lamprecht, Dirk A. ; Steyn, Adrie J.C. ; Ioerger, Thomas R. ; Sacchettini, Jim ; Rhee, Kyu Y. ; Arora, Kriti ; Barry, Clifton E. ; Wyatt, Paul G. ; Boshoff, Helena I.M. / 2-Mercapto-Quinazolinones as Inhibitors of Type II NADH Dehydrogenase and Mycobacterium tuberculosis : Structure-Activity Relationships, Mechanism of Action and Absorption, Distribution, Metabolism, and Excretion Characterization. In: ACS Infectious Diseases. 2018 ; Vol. 4, No. 6. pp. 954-969.
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abstract = "Mycobacterium tuberculosis (MTb) possesses two nonproton pumping type II NADH dehydrogenase (NDH-2) enzymes which are predicted to be jointly essential for respiratory metabolism. Furthermore, the structure of a closely related bacterial NDH-2 has been reported recently, allowing for the structure-based design of small-molecule inhibitors. Herein, we disclose MTb whole-cell structure-activity relationships (SARs) for a series of 2-mercapto-quinazolinones which target the ndh encoded NDH-2 with nanomolar potencies. The compounds were inactivated by glutathione-dependent adduct formation as well as quinazolinone oxidation in microsomes. Pharmacokinetic studies demonstrated modest bioavailability and compound exposures. Resistance to the compounds in MTb was conferred by promoter mutations in the alternative nonessential NDH-2 encoded by ndhA in MTb. Bioenergetic analyses revealed a decrease in oxygen consumption rates in response to inhibitor in cells in which membrane potential was uncoupled from ATP production, while inverted membrane vesicles showed mercapto-quinazolinone-dependent inhibition of ATP production when NADH was the electron donor to the respiratory chain. Enzyme kinetic studies further demonstrated noncompetitive inhibition, suggesting binding of this scaffold to an allosteric site. In summary, while the initial MTb SAR showed limited improvement in potency, these results, combined with structural information on the bacterial protein, will aid in the future discovery of new and improved NDH-2 inhibitors.",
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2-Mercapto-Quinazolinones as Inhibitors of Type II NADH Dehydrogenase and Mycobacterium tuberculosis : Structure-Activity Relationships, Mechanism of Action and Absorption, Distribution, Metabolism, and Excretion Characterization. / Murugesan, Dinakaran; Ray, Peter C.; Bayliss, Tracy; Prosser, Gareth A.; Harrison, Justin R.; Green, Kirsteen; Soares De Melo, Candice; Feng, Tzu Shean; Street, Leslie J.; Chibale, Kelly; Warner, Digby F.; Mizrahi, Valerie; Epemolu, Ola; Scullion, Paul; Ellis, Lucy; Riley, Jennifer; Shishikura, Yoko; Ferguson, Liam; Osuna-Cabello, Maria; Read, Kevin D.; Green, Simon R.; Lamprecht, Dirk A.; Steyn, Adrie J.C.; Ioerger, Thomas R.; Sacchettini, Jim; Rhee, Kyu Y.; Arora, Kriti; Barry, Clifton E.; Wyatt, Paul G.; Boshoff, Helena I.M.

In: ACS Infectious Diseases, Vol. 4, No. 6, 08.06.2018, p. 954-969.

Research output: Contribution to journalArticle

TY - JOUR

T1 - 2-Mercapto-Quinazolinones as Inhibitors of Type II NADH Dehydrogenase and Mycobacterium tuberculosis

T2 - ACS Infectious Diseases

AU - Murugesan, Dinakaran

AU - Ray, Peter C.

AU - Bayliss, Tracy

AU - Prosser, Gareth A.

AU - Harrison, Justin R.

AU - Green, Kirsteen

AU - Soares De Melo, Candice

AU - Feng, Tzu Shean

AU - Street, Leslie J.

AU - Chibale, Kelly

AU - Warner, Digby F.

AU - Mizrahi, Valerie

AU - Epemolu, Ola

AU - Scullion, Paul

AU - Ellis, Lucy

AU - Riley, Jennifer

AU - Shishikura, Yoko

AU - Ferguson, Liam

AU - Osuna-Cabello, Maria

AU - Read, Kevin D.

AU - Green, Simon R.

AU - Lamprecht, Dirk A.

AU - Steyn, Adrie J.C.

AU - Ioerger, Thomas R.

AU - Sacchettini, Jim

AU - Rhee, Kyu Y.

AU - Arora, Kriti

AU - Barry, Clifton E.

AU - Wyatt, Paul G.

AU - Boshoff, Helena I.M.

PY - 2018/6/8

Y1 - 2018/6/8

N2 - Mycobacterium tuberculosis (MTb) possesses two nonproton pumping type II NADH dehydrogenase (NDH-2) enzymes which are predicted to be jointly essential for respiratory metabolism. Furthermore, the structure of a closely related bacterial NDH-2 has been reported recently, allowing for the structure-based design of small-molecule inhibitors. Herein, we disclose MTb whole-cell structure-activity relationships (SARs) for a series of 2-mercapto-quinazolinones which target the ndh encoded NDH-2 with nanomolar potencies. The compounds were inactivated by glutathione-dependent adduct formation as well as quinazolinone oxidation in microsomes. Pharmacokinetic studies demonstrated modest bioavailability and compound exposures. Resistance to the compounds in MTb was conferred by promoter mutations in the alternative nonessential NDH-2 encoded by ndhA in MTb. Bioenergetic analyses revealed a decrease in oxygen consumption rates in response to inhibitor in cells in which membrane potential was uncoupled from ATP production, while inverted membrane vesicles showed mercapto-quinazolinone-dependent inhibition of ATP production when NADH was the electron donor to the respiratory chain. Enzyme kinetic studies further demonstrated noncompetitive inhibition, suggesting binding of this scaffold to an allosteric site. In summary, while the initial MTb SAR showed limited improvement in potency, these results, combined with structural information on the bacterial protein, will aid in the future discovery of new and improved NDH-2 inhibitors.

AB - Mycobacterium tuberculosis (MTb) possesses two nonproton pumping type II NADH dehydrogenase (NDH-2) enzymes which are predicted to be jointly essential for respiratory metabolism. Furthermore, the structure of a closely related bacterial NDH-2 has been reported recently, allowing for the structure-based design of small-molecule inhibitors. Herein, we disclose MTb whole-cell structure-activity relationships (SARs) for a series of 2-mercapto-quinazolinones which target the ndh encoded NDH-2 with nanomolar potencies. The compounds were inactivated by glutathione-dependent adduct formation as well as quinazolinone oxidation in microsomes. Pharmacokinetic studies demonstrated modest bioavailability and compound exposures. Resistance to the compounds in MTb was conferred by promoter mutations in the alternative nonessential NDH-2 encoded by ndhA in MTb. Bioenergetic analyses revealed a decrease in oxygen consumption rates in response to inhibitor in cells in which membrane potential was uncoupled from ATP production, while inverted membrane vesicles showed mercapto-quinazolinone-dependent inhibition of ATP production when NADH was the electron donor to the respiratory chain. Enzyme kinetic studies further demonstrated noncompetitive inhibition, suggesting binding of this scaffold to an allosteric site. In summary, while the initial MTb SAR showed limited improvement in potency, these results, combined with structural information on the bacterial protein, will aid in the future discovery of new and improved NDH-2 inhibitors.

KW - mercapto-quinazolinones

KW - Mycobacterium tuberculosis

KW - respiration

KW - small molecule NDH-2 inhibitors

KW - structure-activity relationship

KW - type II NADH dehydrogenase

U2 - 10.1021/acsinfecdis.7b00275

DO - 10.1021/acsinfecdis.7b00275

M3 - Article

VL - 4

SP - 954

EP - 969

JO - ACS Infectious Diseases

JF - ACS Infectious Diseases

SN - 2373-8227

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