Simple and rapid determination of the enzyme kinetics of HIV-1 reverse transcriptase and anti-HIV-1 agents by a fluorescence based method

Silprasit, Kun; Thammaporn, Ratsupa; Tecchasakul, Supanna; Hannongbua, Supa; Choowongkomon, Kiattawee

doi:10.1016/j.jviromet.2010.12.001

Cited by 23 publications

(23 citation statements)

References 22 publications

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“…Different units (0.1, 0.2, 0.3, 0.4, 0.5 U) of the HIV‐1 RT at a final volume of 5 μ l were added to the reaction mixture for plotting the standard curve. The reaction samples were incubated at 25 °C for 30 min and stopped by adding 2 μ L of 200 m m EDTA . The inhibitory activity of sporolide B was compared with that of the known HIV‐1 RT inhibitor, nevirapine.…”

Section: Methodsmentioning

confidence: 99%

Biological Activity of Sporolides A and B from Salinispora tropica: in silico Target Prediction Using Ligand‐Based Pharmacophore Mapping and in vitro activity Validation on HIV‐1 Reverse Transcriptase

et al. 2014

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Sporolides A and B are novel polycyclic macrolides from the obligate marine actinomycetes, Salinispora tropica. The unique and novel structure of sporolides makes them interesting candidates for targeting diverse biological activities. Biological target prediction of sporolides was carried out using ligand-based pharmacophore screening against known inhibitors and drugs. Validation of pharmacophore screening was carried out for the identified hits. New biological targets predicted for sporolides using this method were HIV-1 reverse transcriptase, adenosine A3 receptor, endothelin receptor ET-A, oxytocin receptor, voltage-gated L-type calcium channel α-1C subunit/calcium channel α/Δ subunit 1. Drug-likeness properties were predicted for the selected compounds using QikProp module. Sporolides A and B showed maximum docking score with HIV-1 reverse transcriptase. Structural interaction fingerprints analysis indicated similar binding pattern of the sporolides with the HIV-1 reverse transcriptase. Sporolide B exhibited good inhibitory activity against HIV-1 reverse transcriptase in in vitro fluorescent assay.

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Section: Methodsmentioning

confidence: 99%

Biological Activity of Sporolides A and B from Salinispora tropica: in silico Target Prediction Using Ligand‐Based Pharmacophore Mapping and in vitro activity Validation on HIV‐1 Reverse Transcriptase

et al. 2014

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“…Anti-HIV-1 reverse transcriptase assay: Anti-HIV-1 reverse transcriptase assay extracts were dissolved in DMSO at the concentration of 20 mg/mL and processed to remove tannin [22]. The assay was carried out in duplicate in a 96-well culture plate using tannin-free supernatant of the extracts as previously described [23]. An appropriate amount of HIV-1 reverse transcriptase (Amersham Pharmacia Biotech Asia Pacific Ltd., Hong Kong) was standardized with fagaronine chloride.…”

Section: Anti-hivs Assaymentioning

confidence: 99%

Cancer Cytotoxic and Anti-HIV Potential of Triphala Herb Mixture on from Chae Son, Lampang, Thailand

Pompimon

Sombutsiri

Baison

et al. 2019

JPRI

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Introduction: The objective of this study was to compare the stability properties of the two years for the quality of Tri-Pha-La Herbal Drink, Thai traditional and Ayurvedic medicine. This study aimed to compare the anticancer and anti-HIV efficacy of the solvent extracts and gallic acid of the three fruits. The research was done between A.D. 2017 to A.D. 2018. Materials and Methods: In the present study, the cancer cell lines, KB, HT 29, MCF-7, A 549, CL and ASK treated with Triphala fruits extract at concentrations of 4 mg/mL for 72 h except 48 for P-388) . The cytotoxic activity was expressed as 50% effective dose (ED50). In addition, anti-HIV activity of the all Triphala fruits solvent extracts was performed by anti-HIVs-1RT. The tested extracts were prescreened at 200 μg/mL and only those which were very active (≥70% inhibition) at this concentration were further determined for the dose, μM, that inhibited 50% HIV-1 RT activity (IC50). Moreover, anti-syncytium (MC99+1A2) was performed by the Triphala fruits extract. The results were expressed as the concentration that inhibited 50% formazan formation in uninfected cells (IC50). The therapeutic index (TI) was calculated using the equation: TI=IC50/EC50. Results: The results were compared between years A.D. 2017/2018. Amongst six ethyl acetate and methanol extracts (A.D. 2017) showed less than 20 µg/mL cell viability in P-388 cell lines. However, the four ethyl acetate and methanol extracts (A.D. 2018) were also inhibited P-388 cell species. Furthermore, each gallic acid, isolated from three fruits Triphala (A.D.2017/2018) were also strongly inhibited to P-388 cell species. Anti-HIV-RT activity of the all Triphala fruits solvent extracts (A.D. 2017) except hexane extracts showed very active which is consistent with A.D. 2018. Conclusion: P. emblica-ethyl acetate extract (A.D. 2018) was the most promising anti-syncytium (MC99+1A2) among evaluated all three fruits Triphala extracts: it afforded the lowest EC ₅₀ 8.85 µM and therapeutic index 2.10. The results evidently showed the capacity of Triphala as potential chemopreventive and anticancer drug and immunity.

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“…The HIV-1 reverse transcriptase inhibition assay (anti-HIV-1 RT) assay followed the fluorescence method from Silprasit et al (2011) with modifications. A reverse transcriptase assay kit was purchased from EnzChek ® , Molecular Probes, Inc., the Netherlands.…”

Section: Hiv-1 Reverse Transcriptase Inhibition Assay (Anti-hiv-1 Rt)mentioning

confidence: 99%

“…The fluorescence of the reaction was measured in a microplate reader with an excitation wavelength of 502 nm and an emission wavelength of 523 nm. The inhibitory effect on HIV-1 RT activity was expressed as the percentage of relative inhibition, using Equation 2 (Silprasit et al, 2011):…”

Section: Hiv-1 Reverse Transcriptase Inhibition Assay (Anti-hiv-1 Rt)mentioning

confidence: 99%

Antioxidant and anti-HIV properties and isolation of bioactive compound of Hymenodictyon orixense (Roxb.) Mabb.

2019

ANRES

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Hymenodictyon orixense (Roxb.) Mabb., or "Som Kob" or "U-Lok" in Thai, is known as a folk medicinal plant. This plant has been used against different dermal diseases, fever and chicken pox, indicating different bioactivity potentials. Methanolic extracts from three parts (bark, wood, fruit) of H. orixense were tested for their antioxidant activities using five assays (total phenolic content, total flavonoid content, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, ferric reducing antioxidant power, nitric oxide radical activity). Bark crude extract had the highest antioxidant activity, so this part was chosen for further fractionation. The fractions were ranked based on from antioxidant and anti-HIV bioactivities. Fraction 2 had the highest level for total flavonoid content (323.63 mg of quercetin (QE) per gram) and nitric oxide radical activity (half maximal inhibitory concentration (IC 50); 88.38 µg/mL). Fraction 4 contained the highest amounts of phenolic compounds (32.89 mg of gallic acid equivalent/g), while Fraction 5 had the highest activity of DPPH radical activity (IC 50 ; 192.76 µg/mL) and ferric reducing antioxidant power (88.08 mg of trolox equivalent/g). For anti-HIV activity, all fractions produced a percentage inhibition greater than 50%, with Fraction 3 being the highest (89.80%) compared to the positive control (97.06%). In addition, scopoletin was isolated from the chloroform extract of the H. orixense bark. Isolation and structural elucidation of other crude chloroform extracts will be further analyzed. The current study demonstrated the potential of developing H. orixense as an antioxidant and anti-HIV agent.

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Simple and rapid determination of the enzyme kinetics of HIV-1 reverse transcriptase and anti-HIV-1 agents by a fluorescence based method

Cited by 23 publications

References 22 publications

Biological Activity of Sporolides A and B from Salinispora tropica: in silico Target Prediction Using Ligand‐Based Pharmacophore Mapping and in vitro activity Validation on HIV‐1 Reverse Transcriptase

Biological Activity of Sporolides A and B from Salinispora tropica: in silico Target Prediction Using Ligand‐Based Pharmacophore Mapping and in vitro activity Validation on HIV‐1 Reverse Transcriptase

Cancer Cytotoxic and Anti-HIV Potential of Triphala Herb Mixture on from Chae Son, Lampang, Thailand

Antioxidant and anti-HIV properties and isolation of bioactive compound of Hymenodictyon orixense (Roxb.) Mabb.

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