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Inhibition of HIV-1 by curcumin A, a novel curcumin analog.

Kumari N, Kulkarni AA, Lin X, McLean C, Ammosova T, Ivanov A, Hipolito M, Nekhai S, Nwulia E - Drug Des Devel Ther (2015)

Bottom Line: The lead compound derived, curcumin A, showed increased stability, especially in murine serum where it was stable for up to 25 hours, as compared to curcumin that only had a half-life of 10 hours.But in primary peripheral blood mononuclear cells, curcumin A inhibited HIV-1 more potently (IC50=2 μM) compared to curcumin (IC50=12 μM).Analysis of specific steps of HIV-1 replication showed that curcumin A inhibited HIV-1 reverse transcription, but had no effect on HIV-1 long terminal repeat basal or Tat-induced transcription, or NF-κB-driven transcription at low concentrations that affected reverse transcription.

View Article: PubMed Central - PubMed

Affiliation: Translational Neuroscience Laboratory, Howard University, Washington, DC, USA ; Department of Medicine, Center for Sickle Cell Disease, College of Medicine, Howard University, Washington, DC, USA.

ABSTRACT
Despite the remarkable success of combination antiretroviral therapy at curtailing HIV progression, emergence of drug-resistant viruses, chronic low-grade inflammation, and adverse effects of combination antiretroviral therapy treatments, including metabolic disorders collectively present the impetus for development of newer and safer antiretroviral drugs. Curcumin, a phytochemical compound, was previously reported to have some in vitro anti-HIV and anti-inflammatory activities, but poor bioavailability has limited its clinical utility. To circumvent the bioavailability problem, we derivatized curcumin to sustain retro-aldol decomposition at physiological pH. The lead compound derived, curcumin A, showed increased stability, especially in murine serum where it was stable for up to 25 hours, as compared to curcumin that only had a half-life of 10 hours. Both curcumin and curcumin A showed similar inhibition of one round of HIV-1 infection in cultured lymphoblastoid (also called CEM) T cells (IC50=0.7 μM). But in primary peripheral blood mononuclear cells, curcumin A inhibited HIV-1 more potently (IC50=2 μM) compared to curcumin (IC50=12 μM). Analysis of specific steps of HIV-1 replication showed that curcumin A inhibited HIV-1 reverse transcription, but had no effect on HIV-1 long terminal repeat basal or Tat-induced transcription, or NF-κB-driven transcription at low concentrations that affected reverse transcription. Finally, we showed curcumin A induced expression of HO-1 and decreased cell cycle progression of T cells. Our findings thus indicate that altering the core structure of curcumin could yield more stable compounds with potent antiretroviral and anti-inflammatory activities.

No MeSH data available.


Related in: MedlinePlus

Effect of curcumin and curcumin A on one round of HIV-1-Luc replication and cellular viability in CEM-T cells and PBMCs.Notes: CEM-T cells (A) or peripheral blood mononuclear cells (PBMCs) activated with PHA and IL-2 (C) were infected with VSVG-pseudotyped pNL4-3.Luc.R-E- (HIV-1 Luc) virus and treated with the indicated concentrations of curcumin or curcumin A for 48 hours at 37°C; then the cells were lyzed and luciferase activity was measured. (B) CEM-T cells were treated with the indicated concentrations of curcumin or curcumin A for 48 hours at 37°C. CEM-T cells were treated with 0.4 μM calcein-AM for 30 minutes and calcein fluorescence was measured at 485 nm excitation and 515 nm emission on the luminescence spectrometer equipped with the robotic arm (PerkinElmer LS 50B; PerkinElmer Inc., Waltham, MA, USA). (D) Activated PBMCs were treated with the indicated concentrations of curcumin or curcumin A for 48 hours at 37°C and viability of cells was measured by trypan blue exclusion method. IC50s shown on panels (A–D) were determined with GraphPad Prism 6 Software (GraphPad Software, Inc., La Jolla, CA, USA).Abbreviations: PHA, phytohemagglutinin; VSVG, vesicular stomatitis virus glycoprotein G; IC50, the half maximal inhibitory concentration; M, molar concentration.
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f3-dddt-9-5051: Effect of curcumin and curcumin A on one round of HIV-1-Luc replication and cellular viability in CEM-T cells and PBMCs.Notes: CEM-T cells (A) or peripheral blood mononuclear cells (PBMCs) activated with PHA and IL-2 (C) were infected with VSVG-pseudotyped pNL4-3.Luc.R-E- (HIV-1 Luc) virus and treated with the indicated concentrations of curcumin or curcumin A for 48 hours at 37°C; then the cells were lyzed and luciferase activity was measured. (B) CEM-T cells were treated with the indicated concentrations of curcumin or curcumin A for 48 hours at 37°C. CEM-T cells were treated with 0.4 μM calcein-AM for 30 minutes and calcein fluorescence was measured at 485 nm excitation and 515 nm emission on the luminescence spectrometer equipped with the robotic arm (PerkinElmer LS 50B; PerkinElmer Inc., Waltham, MA, USA). (D) Activated PBMCs were treated with the indicated concentrations of curcumin or curcumin A for 48 hours at 37°C and viability of cells was measured by trypan blue exclusion method. IC50s shown on panels (A–D) were determined with GraphPad Prism 6 Software (GraphPad Software, Inc., La Jolla, CA, USA).Abbreviations: PHA, phytohemagglutinin; VSVG, vesicular stomatitis virus glycoprotein G; IC50, the half maximal inhibitory concentration; M, molar concentration.

Mentions: The effects of curcumin and curcumin A on one round of HIV-1 infection were first analyzed in cultured CEM-T cells infected with VSVG-pseudotyped HIV-1 pNL4-3 virus expressing luciferase in place of nef (HIV-1 Luc).34 The cells were treated with the compounds for 24 hours and luciferase activity was measured as an indicator of HIV-1 replication. One round HIV-1 infection was equally inhibited by curcumin (the half maximal inhibitory concentration [IC50] =0.7 μM) and curcumin A (IC50=0.8 μM) (Figure 3A). Viability of CEM-T cells was analyzed using calcein-AM assay which measured fluorescence of the calcein converted intracellularly from the non-fluorescent calcein-AM taken up by the cells. Curcumin and curcumin A reduced viability of CEM-T cells when added to the cells for 24 hours, with curcumin being slightly more toxic (IC50=1.26 μM) than curcumin A (IC50=2.4 μM) (Figure 3B). We next analyzed the effect of curcumin and curcumin A on single round HIV-1 infection in primary PBMCs. PBMCs were activated by treatment with PHA and IL-2 (see Materials and methods for details), infected with HIV-1 Luc for 24 hours and then treated with curcumin or curcumin A for another 24 hours. Both curcumin and curcumin A markedly inhibited HIV-1 infection with curcumin A being more potent (IC50=2 μM) than curcumin (IC50=12 μM) (Figure 3C). Both curcumin and curcumin A reduced viability of PBMCs as analyzed using trypan blue, with IC50s of 35 μM and 22 μM, respectively (Figure 3D). Thus, curcumin A inhibits one round HIV-1 infection comparable to curcumin in CEM-T cells and displays more potent activity in primary PBMCs. Also, curcumin A demonstrated a better therapeutic window in both cultured CEM-T cells and PBMCs.


Inhibition of HIV-1 by curcumin A, a novel curcumin analog.

Kumari N, Kulkarni AA, Lin X, McLean C, Ammosova T, Ivanov A, Hipolito M, Nekhai S, Nwulia E - Drug Des Devel Ther (2015)

Effect of curcumin and curcumin A on one round of HIV-1-Luc replication and cellular viability in CEM-T cells and PBMCs.Notes: CEM-T cells (A) or peripheral blood mononuclear cells (PBMCs) activated with PHA and IL-2 (C) were infected with VSVG-pseudotyped pNL4-3.Luc.R-E- (HIV-1 Luc) virus and treated with the indicated concentrations of curcumin or curcumin A for 48 hours at 37°C; then the cells were lyzed and luciferase activity was measured. (B) CEM-T cells were treated with the indicated concentrations of curcumin or curcumin A for 48 hours at 37°C. CEM-T cells were treated with 0.4 μM calcein-AM for 30 minutes and calcein fluorescence was measured at 485 nm excitation and 515 nm emission on the luminescence spectrometer equipped with the robotic arm (PerkinElmer LS 50B; PerkinElmer Inc., Waltham, MA, USA). (D) Activated PBMCs were treated with the indicated concentrations of curcumin or curcumin A for 48 hours at 37°C and viability of cells was measured by trypan blue exclusion method. IC50s shown on panels (A–D) were determined with GraphPad Prism 6 Software (GraphPad Software, Inc., La Jolla, CA, USA).Abbreviations: PHA, phytohemagglutinin; VSVG, vesicular stomatitis virus glycoprotein G; IC50, the half maximal inhibitory concentration; M, molar concentration.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4562762&req=5

f3-dddt-9-5051: Effect of curcumin and curcumin A on one round of HIV-1-Luc replication and cellular viability in CEM-T cells and PBMCs.Notes: CEM-T cells (A) or peripheral blood mononuclear cells (PBMCs) activated with PHA and IL-2 (C) were infected with VSVG-pseudotyped pNL4-3.Luc.R-E- (HIV-1 Luc) virus and treated with the indicated concentrations of curcumin or curcumin A for 48 hours at 37°C; then the cells were lyzed and luciferase activity was measured. (B) CEM-T cells were treated with the indicated concentrations of curcumin or curcumin A for 48 hours at 37°C. CEM-T cells were treated with 0.4 μM calcein-AM for 30 minutes and calcein fluorescence was measured at 485 nm excitation and 515 nm emission on the luminescence spectrometer equipped with the robotic arm (PerkinElmer LS 50B; PerkinElmer Inc., Waltham, MA, USA). (D) Activated PBMCs were treated with the indicated concentrations of curcumin or curcumin A for 48 hours at 37°C and viability of cells was measured by trypan blue exclusion method. IC50s shown on panels (A–D) were determined with GraphPad Prism 6 Software (GraphPad Software, Inc., La Jolla, CA, USA).Abbreviations: PHA, phytohemagglutinin; VSVG, vesicular stomatitis virus glycoprotein G; IC50, the half maximal inhibitory concentration; M, molar concentration.
Mentions: The effects of curcumin and curcumin A on one round of HIV-1 infection were first analyzed in cultured CEM-T cells infected with VSVG-pseudotyped HIV-1 pNL4-3 virus expressing luciferase in place of nef (HIV-1 Luc).34 The cells were treated with the compounds for 24 hours and luciferase activity was measured as an indicator of HIV-1 replication. One round HIV-1 infection was equally inhibited by curcumin (the half maximal inhibitory concentration [IC50] =0.7 μM) and curcumin A (IC50=0.8 μM) (Figure 3A). Viability of CEM-T cells was analyzed using calcein-AM assay which measured fluorescence of the calcein converted intracellularly from the non-fluorescent calcein-AM taken up by the cells. Curcumin and curcumin A reduced viability of CEM-T cells when added to the cells for 24 hours, with curcumin being slightly more toxic (IC50=1.26 μM) than curcumin A (IC50=2.4 μM) (Figure 3B). We next analyzed the effect of curcumin and curcumin A on single round HIV-1 infection in primary PBMCs. PBMCs were activated by treatment with PHA and IL-2 (see Materials and methods for details), infected with HIV-1 Luc for 24 hours and then treated with curcumin or curcumin A for another 24 hours. Both curcumin and curcumin A markedly inhibited HIV-1 infection with curcumin A being more potent (IC50=2 μM) than curcumin (IC50=12 μM) (Figure 3C). Both curcumin and curcumin A reduced viability of PBMCs as analyzed using trypan blue, with IC50s of 35 μM and 22 μM, respectively (Figure 3D). Thus, curcumin A inhibits one round HIV-1 infection comparable to curcumin in CEM-T cells and displays more potent activity in primary PBMCs. Also, curcumin A demonstrated a better therapeutic window in both cultured CEM-T cells and PBMCs.

Bottom Line: The lead compound derived, curcumin A, showed increased stability, especially in murine serum where it was stable for up to 25 hours, as compared to curcumin that only had a half-life of 10 hours.But in primary peripheral blood mononuclear cells, curcumin A inhibited HIV-1 more potently (IC50=2 μM) compared to curcumin (IC50=12 μM).Analysis of specific steps of HIV-1 replication showed that curcumin A inhibited HIV-1 reverse transcription, but had no effect on HIV-1 long terminal repeat basal or Tat-induced transcription, or NF-κB-driven transcription at low concentrations that affected reverse transcription.

View Article: PubMed Central - PubMed

Affiliation: Translational Neuroscience Laboratory, Howard University, Washington, DC, USA ; Department of Medicine, Center for Sickle Cell Disease, College of Medicine, Howard University, Washington, DC, USA.

ABSTRACT
Despite the remarkable success of combination antiretroviral therapy at curtailing HIV progression, emergence of drug-resistant viruses, chronic low-grade inflammation, and adverse effects of combination antiretroviral therapy treatments, including metabolic disorders collectively present the impetus for development of newer and safer antiretroviral drugs. Curcumin, a phytochemical compound, was previously reported to have some in vitro anti-HIV and anti-inflammatory activities, but poor bioavailability has limited its clinical utility. To circumvent the bioavailability problem, we derivatized curcumin to sustain retro-aldol decomposition at physiological pH. The lead compound derived, curcumin A, showed increased stability, especially in murine serum where it was stable for up to 25 hours, as compared to curcumin that only had a half-life of 10 hours. Both curcumin and curcumin A showed similar inhibition of one round of HIV-1 infection in cultured lymphoblastoid (also called CEM) T cells (IC50=0.7 μM). But in primary peripheral blood mononuclear cells, curcumin A inhibited HIV-1 more potently (IC50=2 μM) compared to curcumin (IC50=12 μM). Analysis of specific steps of HIV-1 replication showed that curcumin A inhibited HIV-1 reverse transcription, but had no effect on HIV-1 long terminal repeat basal or Tat-induced transcription, or NF-κB-driven transcription at low concentrations that affected reverse transcription. Finally, we showed curcumin A induced expression of HO-1 and decreased cell cycle progression of T cells. Our findings thus indicate that altering the core structure of curcumin could yield more stable compounds with potent antiretroviral and anti-inflammatory activities.

No MeSH data available.


Related in: MedlinePlus