Inhibition of cytomegalovirus replication with extended-half-life synthetic ozonides

Yiping Wang, Rupkatha Mukhopadhyay, Sujayita Roy, Arun Kapoor, Yu Pin Su, Susan A. Charman, Gong Chen, Jianbo Wu, Xiaofang Wang, Jonathan L. Vennerstrom, Ravit Arav-Boger

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Artesunate (AS), a semisynthetic artemisinin approved for malaria therapy, inhibits human cytomegalovirus (HCMV) replication in vitro, but therapeutic success in humans has been variable. We hypothesized that the short in vivo half-life of AS may contribute to the different treatment outcomes. We tested novel synthetic ozonides with longer half-lives against HCMV in vitro and mouse cytomegalovirus (MCMV) in vivo. Screening of the activities of four ozonides against a pp28-luciferase-expressing HCMV Towne recombinant identified OZ418 to have the best selectivity; its effective concentration inhibiting viral growth by 50% (EC50) was 9.8 0.2 M, and cytotoxicity in noninfected human fibroblasts (the concentration inhibiting cell growth by 50% [CC50]) was 128.1 8.0 M. In plaque reduction assays, OZ418 inhibited HCMV TB40 in a concentration-dependent manner as well as a ganciclovir (GCV)-resistant HCMV isolate. The combination of OZ418 and GCV was synergistic in HCMV inhibition in vitro. Virus inhibition by OZ418 occurred at an early stage and was dependent on the cell density at the time of infection. OZ418 treatment reversed HCMV-mediated cell cycle progression and correlated with the reduction of HCMV-induced expression of pRb, E2F1, and cyclin-dependent kinases 1, 2, 4, and 6. In an MCMV model, once-daily oral administration of OZ418 had significantly improved efficacy against MCMV compared to that of twice-daily oral AS. A parallel pharmacokinetic study with a single oral dose of OZ418 or AS showed a prolonged plasma half-life and higher unbound concentrations of OZ418 than unbound concentrations of AS. In summary, ozonides are proposed to be potential therapeutics, alone or in combination with GCV, for HCMV infection in humans.

Original languageEnglish
Article numbere01735-18
Number of pages14
JournalAntimicrobial Agents and Chemotherapy
Volume63
Issue number1
DOIs
Publication statusPublished - 1 Jan 2019

Keywords

  • Artemisinins
  • Half-life
  • Human cytomegalovirus
  • Mouse cytomegalovirus
  • Ozonides
  • Pharmacokinetics

Cite this

Wang, Yiping ; Mukhopadhyay, Rupkatha ; Roy, Sujayita ; Kapoor, Arun ; Su, Yu Pin ; Charman, Susan A. ; Chen, Gong ; Wu, Jianbo ; Wang, Xiaofang ; Vennerstrom, Jonathan L. ; Arav-Boger, Ravit. / Inhibition of cytomegalovirus replication with extended-half-life synthetic ozonides. In: Antimicrobial Agents and Chemotherapy. 2019 ; Vol. 63, No. 1.
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title = "Inhibition of cytomegalovirus replication with extended-half-life synthetic ozonides",
abstract = "Artesunate (AS), a semisynthetic artemisinin approved for malaria therapy, inhibits human cytomegalovirus (HCMV) replication in vitro, but therapeutic success in humans has been variable. We hypothesized that the short in vivo half-life of AS may contribute to the different treatment outcomes. We tested novel synthetic ozonides with longer half-lives against HCMV in vitro and mouse cytomegalovirus (MCMV) in vivo. Screening of the activities of four ozonides against a pp28-luciferase-expressing HCMV Towne recombinant identified OZ418 to have the best selectivity; its effective concentration inhibiting viral growth by 50{\%} (EC50) was 9.8 0.2 M, and cytotoxicity in noninfected human fibroblasts (the concentration inhibiting cell growth by 50{\%} [CC50]) was 128.1 8.0 M. In plaque reduction assays, OZ418 inhibited HCMV TB40 in a concentration-dependent manner as well as a ganciclovir (GCV)-resistant HCMV isolate. The combination of OZ418 and GCV was synergistic in HCMV inhibition in vitro. Virus inhibition by OZ418 occurred at an early stage and was dependent on the cell density at the time of infection. OZ418 treatment reversed HCMV-mediated cell cycle progression and correlated with the reduction of HCMV-induced expression of pRb, E2F1, and cyclin-dependent kinases 1, 2, 4, and 6. In an MCMV model, once-daily oral administration of OZ418 had significantly improved efficacy against MCMV compared to that of twice-daily oral AS. A parallel pharmacokinetic study with a single oral dose of OZ418 or AS showed a prolonged plasma half-life and higher unbound concentrations of OZ418 than unbound concentrations of AS. In summary, ozonides are proposed to be potential therapeutics, alone or in combination with GCV, for HCMV infection in humans.",
keywords = "Artemisinins, Half-life, Human cytomegalovirus, Mouse cytomegalovirus, Ozonides, Pharmacokinetics",
author = "Yiping Wang and Rupkatha Mukhopadhyay and Sujayita Roy and Arun Kapoor and Su, {Yu Pin} and Charman, {Susan A.} and Gong Chen and Jianbo Wu and Xiaofang Wang and Vennerstrom, {Jonathan L.} and Ravit Arav-Boger",
year = "2019",
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doi = "10.1128/AAC.01735-18",
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Wang, Y, Mukhopadhyay, R, Roy, S, Kapoor, A, Su, YP, Charman, SA, Chen, G, Wu, J, Wang, X, Vennerstrom, JL & Arav-Boger, R 2019, 'Inhibition of cytomegalovirus replication with extended-half-life synthetic ozonides' Antimicrobial Agents and Chemotherapy, vol. 63, no. 1, e01735-18. https://doi.org/10.1128/AAC.01735-18

Inhibition of cytomegalovirus replication with extended-half-life synthetic ozonides. / Wang, Yiping; Mukhopadhyay, Rupkatha; Roy, Sujayita; Kapoor, Arun; Su, Yu Pin; Charman, Susan A.; Chen, Gong; Wu, Jianbo; Wang, Xiaofang; Vennerstrom, Jonathan L.; Arav-Boger, Ravit.

In: Antimicrobial Agents and Chemotherapy, Vol. 63, No. 1, e01735-18, 01.01.2019.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Inhibition of cytomegalovirus replication with extended-half-life synthetic ozonides

AU - Wang, Yiping

AU - Mukhopadhyay, Rupkatha

AU - Roy, Sujayita

AU - Kapoor, Arun

AU - Su, Yu Pin

AU - Charman, Susan A.

AU - Chen, Gong

AU - Wu, Jianbo

AU - Wang, Xiaofang

AU - Vennerstrom, Jonathan L.

AU - Arav-Boger, Ravit

PY - 2019/1/1

Y1 - 2019/1/1

N2 - Artesunate (AS), a semisynthetic artemisinin approved for malaria therapy, inhibits human cytomegalovirus (HCMV) replication in vitro, but therapeutic success in humans has been variable. We hypothesized that the short in vivo half-life of AS may contribute to the different treatment outcomes. We tested novel synthetic ozonides with longer half-lives against HCMV in vitro and mouse cytomegalovirus (MCMV) in vivo. Screening of the activities of four ozonides against a pp28-luciferase-expressing HCMV Towne recombinant identified OZ418 to have the best selectivity; its effective concentration inhibiting viral growth by 50% (EC50) was 9.8 0.2 M, and cytotoxicity in noninfected human fibroblasts (the concentration inhibiting cell growth by 50% [CC50]) was 128.1 8.0 M. In plaque reduction assays, OZ418 inhibited HCMV TB40 in a concentration-dependent manner as well as a ganciclovir (GCV)-resistant HCMV isolate. The combination of OZ418 and GCV was synergistic in HCMV inhibition in vitro. Virus inhibition by OZ418 occurred at an early stage and was dependent on the cell density at the time of infection. OZ418 treatment reversed HCMV-mediated cell cycle progression and correlated with the reduction of HCMV-induced expression of pRb, E2F1, and cyclin-dependent kinases 1, 2, 4, and 6. In an MCMV model, once-daily oral administration of OZ418 had significantly improved efficacy against MCMV compared to that of twice-daily oral AS. A parallel pharmacokinetic study with a single oral dose of OZ418 or AS showed a prolonged plasma half-life and higher unbound concentrations of OZ418 than unbound concentrations of AS. In summary, ozonides are proposed to be potential therapeutics, alone or in combination with GCV, for HCMV infection in humans.

AB - Artesunate (AS), a semisynthetic artemisinin approved for malaria therapy, inhibits human cytomegalovirus (HCMV) replication in vitro, but therapeutic success in humans has been variable. We hypothesized that the short in vivo half-life of AS may contribute to the different treatment outcomes. We tested novel synthetic ozonides with longer half-lives against HCMV in vitro and mouse cytomegalovirus (MCMV) in vivo. Screening of the activities of four ozonides against a pp28-luciferase-expressing HCMV Towne recombinant identified OZ418 to have the best selectivity; its effective concentration inhibiting viral growth by 50% (EC50) was 9.8 0.2 M, and cytotoxicity in noninfected human fibroblasts (the concentration inhibiting cell growth by 50% [CC50]) was 128.1 8.0 M. In plaque reduction assays, OZ418 inhibited HCMV TB40 in a concentration-dependent manner as well as a ganciclovir (GCV)-resistant HCMV isolate. The combination of OZ418 and GCV was synergistic in HCMV inhibition in vitro. Virus inhibition by OZ418 occurred at an early stage and was dependent on the cell density at the time of infection. OZ418 treatment reversed HCMV-mediated cell cycle progression and correlated with the reduction of HCMV-induced expression of pRb, E2F1, and cyclin-dependent kinases 1, 2, 4, and 6. In an MCMV model, once-daily oral administration of OZ418 had significantly improved efficacy against MCMV compared to that of twice-daily oral AS. A parallel pharmacokinetic study with a single oral dose of OZ418 or AS showed a prolonged plasma half-life and higher unbound concentrations of OZ418 than unbound concentrations of AS. In summary, ozonides are proposed to be potential therapeutics, alone or in combination with GCV, for HCMV infection in humans.

KW - Artemisinins

KW - Half-life

KW - Human cytomegalovirus

KW - Mouse cytomegalovirus

KW - Ozonides

KW - Pharmacokinetics

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