1.Studies on the inhibition of polyhydroxylated aromatic compounds against HIV-1 integrase.
Zhi-min GUO ; Hong-shan CHEN ; Lin WANG
Acta Pharmaceutica Sinica 2002;37(4):253-256
AIMThree major enzymes of HIV-1, reverse transcriptase (RT), protease (PR), and integrase (IN), are important targets for anti-HIV drugs. Nine RT and five PR inhibitors have been effectively used in treatment of AIDS patients. In order to find active integrase inhibitors, twenty polyhydroxylated aromatic compounds were tested.
METHODSELISA method was used to test the integrase activity. The synthesized donor substrate oligonucleotide representing the HIV-1 U5LTR was immobilized onto Covalink polystyrene microtiter plates, and a synthesized biotinlated 20 bp oligonucleotide was used as the target substrate. The products were detected and quantified by a colorimetric avidin-linked alkaline phosphatase reporter system.
RESULTSCompound NQ-2 was found to inhibit HIV-1 integrase with the IC50 of 78.5 mumol.L-1 by ELISA method. Its novel analogue NQ-3 was found to be 2 fold more potent on HIV intrgrase than NQ-2, IC50 was 37.2 mumol.L-1. The IC50s of NQ-2 and NQ-3 to inhibit the 3'-pro + assembly activity of integrase were 96.94 mumol.L-1 and 8.48 mumol.L-1; to inhibit assembly activity were 168 and 6.9 mumol.L-1 and to inhibit strand-transfer activity were 49.8 and 1.1 mumol.L-1, respectively. Compound NQ-2 mostly inhibited the strand transfer activity of HIV-1 integrase. Compound NQ-3 inhibited both the assembly and strand-transfer with high activities.
CONCLUSIONNaphthoquinone compound NQ-3 was found to be a novel HIV integrase inhibitor which warrants further study. Uncoupled ELISA HIV integrase assay is shown to be useful to screen HIV-1 integrase inhibitors.
Coumarins ; pharmacology ; HIV Integrase ; drug effects ; metabolism ; HIV Integrase Inhibitors ; pharmacology ; HIV-1 ; drug effects ; enzymology ; Inhibitory Concentration 50 ; Naphthoquinones ; pharmacology ; Polycyclic Aromatic Hydrocarbons ; pharmacology ; Stilbenes ; pharmacology
2.Design, synthesis of quinolinone acid-containing compounds with anti-HIV integrase activity.
Xiao-fang CHEN ; Yan-bin WU ; Jie JIN ; Rui-zhen WANG ; Chong WANG ; Jun LIU
Acta Pharmaceutica Sinica 2010;45(2):263-267
A series of novel quinolinone acid-containing compounds were designed and synthesized. Their structures were confirmed with 1H NMR and MS. The target compounds were tested for anti-HIV-1 integrase activities in vitro with enzyme linked immunosorbent assay (ELISA). The result showed that D-2, D-4 and D-7 have anti-integrase activity with IC50 < 100 micromol L(-1).
HIV Integrase
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metabolism
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HIV Integrase Inhibitors
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chemical synthesis
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chemistry
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pharmacology
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Inhibitory Concentration 50
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Quinolones
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chemical synthesis
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chemistry
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pharmacology
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Structure-Activity Relationship
3.LEDGF/p75: a novel target for anti-HIV therapy and advances in the study of its related inhibitors.
Acta Pharmaceutica Sinica 2009;44(9):953-960
LEDGF/p75 is a newly found cell cofactor, which plays an essential role in the integration of HIV-1 cDNA into host chromosomes. LEDGF/p75 tethers HIV integrase to chromatin, protects it from degradation, and strongly influences the genome-wide pattern of HIV integration. Depleting the protein from cells or over-expressing the integrase-binding domain of LEDGF/p75 blocks viral replication. The essential role of LEDGF/p75 in HIV-1 replication makes it a new target for anti-HIV-1 drug development. This article reviews the function of LEDGF/p75, LEDGF/p75-integrase interaction and LEDGF/p75 inhibitors.
Anti-HIV Agents
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chemistry
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pharmacology
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HIV Integrase
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metabolism
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HIV-1
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drug effects
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physiology
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Intercellular Signaling Peptides and Proteins
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metabolism
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Protein Binding
4.Advances in the study of HIV-1 integrase inhibitors of alpha, gamma-diketo compounds.
Sheng-hui YU ; Yan-mei TAN ; Gui-sen ZHAO
Acta Pharmaceutica Sinica 2010;45(2):215-223
HIV-1 integrase (IN) is an essential enzyme for retroviral replication. There is no analogue for this enzyme in human cells so that inhibition of IN will not bring strong effect on human body. Thus, HIV-1 IN has become a rational target for therapy of AIDS. This review provides a comprehensive report of alpha, gamma-diketo IN inhibitors discovered in recent years. Compilation of such data will prove to be beneficial in developing QSAR, pharmacophore hypothesis generation and validation, virtual screening and synthesis of compounds with higher activity.
Anti-HIV Agents
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chemical synthesis
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chemistry
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pharmacology
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HIV Integrase
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chemistry
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physiology
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HIV Integrase Inhibitors
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chemical synthesis
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chemistry
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pharmacology
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HIV-1
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drug effects
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Humans
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Keto Acids
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chemical synthesis
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chemistry
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pharmacology
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Molecular Structure
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Quantitative Structure-Activity Relationship
5.Inhibitory effect of endophytic fungi from Dysosma versipellis on HIV-1 IN-LEDGF/p75 interaction.
Ya-Qin ZHOU ; Da-Wei ZHANG ; Li-Ying YU ; Ying WEI ; Hong-Zhen TANG ; Shi-Ling YANG ; Xiao-Ming TAN
China Journal of Chinese Materia Medica 2019;44(9):1808-1813
To determine the inhibitory effect of endophytic fungi from Dysosma versipellis on HIV-1 IN-LEDGF/p75 interaction,the protein-protein interaction between human immunodeficiency virus type 1( HIV-1) integrase and lens epithelial growth factor p75 protein( LEDGF/p75) was used as a target. The homogeneous time-resolved fluorescence( HTRF) technique was used in the inhibitory activity assay. The results showed that eight endophytic fungi with anti-IN-LEDGF/p75 interaction activity were screened out from fifty-three strains with different morphological characteristic. Among them,106 strain showed strong inhibitory activity against HIV-1 IN-LEDGF/p75 interaction with IC50 value of 5. 23 mg·L-1,and was identified as a potential novel species of Magnaporthaceae family by the analyses of ITS-rDNA,LSU and RPB2 sequences data. This study demonstrated that potential natural active ingredients against the HIV-1 IN-LEDGF/p75 interaction exist in the endophytic fungi of D. versipellis. These results may provide available candidate strain resources for the research and development of new anti-acquired immunodeficiency syndrome drugs.
Berberidaceae
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microbiology
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Endophytes
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Fungi
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chemistry
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HIV Integrase
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metabolism
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HIV-1
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drug effects
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Humans
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Intercellular Signaling Peptides and Proteins
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metabolism
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Protein Binding
6.HIV-1 integrase enzyme linked immunosorbent assay and inhibitors.
Chinese Journal of Experimental and Clinical Virology 2002;16(2):119-123
BACKGROUNDTo establish an ELISA method for detecting the activity of HIV-1 integrase and screening of inhibitors.
METHODSHIV recombinant plasmid F185K/C280S IN1-288 was transformed into the E.coli strain BL21(DE3) integrase with a histag was induced by adding isopropyl-?-D?thiogalactopyranoside (IPTG)and purified by nickel affinity chromatography. The synthesized donor substrate oligonucleotide representing the HIV-1 U5LTR was immobilized onto covalink polystyrene microtiter plates, and a synthesized biotinlated 20 bp oligonucleotide was used as the target substrate. The products were detected and quantified using a colorimetic avidin?linked alkaline phosphatase reporter system,and identified by 32? autoradiography. Some natural products and chemically synthesized compounds were screened for HIV-1 integrase inhibitors.
RESULTSThe purified integrase was identified by SDS?PAGE and showed integration activity by ELISA and?32P radioisotopic assay.?Coefficients of variation (CV)of ELISA in a lot and among the lots were 4.63% and 5.89% respectively, the mean of P/N was 2.836 0.161 under the optimal experimental condition. Some plant extracts were found as potent integrase inhibitors. The IC50s for CEH and CEHL were (20.41 5.68)?g/ml and (7.56 1.86)?g/ml respectively.
CONCLUSIONSThe authors have established a simple and rapid ELISA method with stable repeatability for detecting integrase activity, which can be used for screening and studying of specific inhibitors of HIV-1 integrase.
Antiviral Agents ; pharmacology ; Drug Evaluation, Preclinical ; Enzyme-Linked Immunosorbent Assay ; methods ; HIV Integrase ; isolation & purification ; metabolism ; HIV Integrase Inhibitors ; pharmacology ; HIV-1 ; enzymology ; Recombinant Fusion Proteins ; isolation & purification ; metabolism
7.Research progress of dual inhibitors targeting HIV-1 reverse transcriptase and integrase.
Hong LIU ; Peng ZHAN ; Xin-Yong LIU
Acta Pharmaceutica Sinica 2013;48(4):466-476
Both reverse transcriptase (RT) and integrase (IN) play crucial roles in the life cycle of HIV-1, which are also key targets in the area of anti-HIV drug research. Reverse transcriptase inhibitors are involved in the most employed drugs used to treat AIDS patients and HIV-infected people, while one of the integrase inhibitors has already been approved by US FDA to appear on the market. Great achievement has been made in the research on both, separately. Recently, much more attention of medicinal chemistry researchers has been attracted to the strategies of multi-target drugs. Compounds with excellent potency against both HIV RT and IN, evidently defined as dual inhibitors targeting both enzymes, have been obtained through considerable significant exploration, which can be classified into two categories according to different strategies. Combinatorial chemistry approach together with high throughput screening methods and multi-target-based virtual screening strategy have been useful tools for identifying selective anti-HIV compounds for long times; Rational drug design based on pharmacophore combination has also led to remarkable results. In this paper, latest progress of both categories in the discovery and structural modification will be covered, with a view to contribute to the career of anti-HIV research.
Drug Design
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HIV Integrase Inhibitors
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chemistry
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pharmacology
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HIV Reverse Transcriptase
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antagonists & inhibitors
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HIV-1
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drug effects
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Humans
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Molecular Structure
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Reverse Transcriptase Inhibitors
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chemistry
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pharmacology
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Structure-Activity Relationship
8.Advances in the study of HIV-1 integrase inhibitor of aryl beta-diketoacids.
Acta Pharmaceutica Sinica 2006;41(9):801-807
Animals
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Anti-HIV Agents
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chemistry
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pharmacology
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Furans
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pharmacology
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HIV Integrase Inhibitors
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chemistry
;
pharmacology
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HIV-1
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drug effects
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Humans
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Keto Acids
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chemistry
;
pharmacology
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Molecular Structure
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Naphthyridines
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pharmacology
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Triazoles
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pharmacology
9.Isolation, idetification and anti-HIV-1 integrase activity of culturable endophytic fungi from Tibetan medicinal plant Phlomis younghusbandii Mukerjee.
Da-Wei ZHANG ; Ming-Ming ZHAO ; Juan CHEN ; Chao LI ; Shun-Xing GUO
Acta Pharmaceutica Sinica 2013;48(5):780-789
A total of 52 endophytic fungi were isolated from roots and stems of Tibetan medicinal plant Phlomis younghusbandii Mukerjee. These fungal isolates were molecularly identified based on ITS sequnces and 28S sequences distributed to 12 genera, including Phoma, Chaetosphaeronema, Fusarium and Leptosphaeria, etc. Among them, the dominant genus was Phoma. Extracts of all strains were evaluated for anti-HIV-1 integrase activity by using soluable integrase expressed in E. coli BL21 (DE3). The results showed that seven samples from five fungal endophytes PHY-24, PHY-38, PHY-40, PHY-51, PHY-53, which belonged to genus Chaetosphaeronema, inhibited strand transfer reaction catalyzed by HIV-1 integrase with IC50 values, of 6.60, 5.20, 2.86, 7.86, 4.47, 4.56 and 3.23 microg x mL(-1) respectively. In conclusion, the endophytic fungi of Phlomis younghusbandii Mukerjee are valuable for further screening anti-HIV-1 integrase agents.
Ascomycota
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enzymology
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isolation & purification
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Chaetomium
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enzymology
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isolation & purification
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Endophytes
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enzymology
;
isolation & purification
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Escherichia coli
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enzymology
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HIV Integrase
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genetics
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metabolism
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HIV Integrase Inhibitors
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pharmacology
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Phlomis
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microbiology
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Phylogeny
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Plant Roots
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microbiology
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Plant Stems
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microbiology
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Plants, Medicinal
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microbiology
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Plasmids
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Recombinant Proteins
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genetics
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metabolism
10.Wikstroelide M potently inhibits HIV replication by targeting reverse transcriptase and integrase nuclear translocation.
Xuan ZHANG ; Sheng-Zhuo HUANG ; Wan-Gang GU ; Liu-Meng YANG ; Huan CHEN ; Chang-Bo ZHENG ; You-Xing ZHAO ; David Chi-Cheong WAN ; Yong-Tang ZHENG
Chinese Journal of Natural Medicines (English Ed.) 2014;12(3):186-193
AIM:
To evaluate the anti-HIV activity and mechanism of action of wikstroelide M, a daphnane diterpene from Daphne acutiloba Rehder (Thymelaeaceae).
METHODS:
The anti-HIV activities of wikstroelide M against different HIV strains were evaluated by cytopathic effect assay and p24 quantification assay with ELISA. The inhibitory effect of wikstroelide M on HIV reverse transcription was analyzed by real-time PCR and ELISA. The effect of wikstroelide M on HIV-1 integrase nuclear translocation was observed with a cell-based imaging assay. The effect of wikstroelide M on LEDGF/p75-IN interaction was assayed by molecular docking.
RESULTS:
Wikstroelide M potently inhibited different HIV-1 strains, including HIV-1IIIB, HIV-1A17, and HIV-19495, induced a cytopathic effect, with EC50 values ranging from 3.81 to 15.65 ng·mL⁻¹. Wikstroelide M also had high inhibitory activities against HIV-2ROD and HIV-2CBL-20-induced cytopathic effects with EC50 values of 18.88 and 31.90 ng·mL⁻¹. The inhibitory activities of wikstroelide M on the three HIV-1 strains were further confirmed by p24 quantification assay, with EC50 values ranging from 15.16 to 35.57 ng·mL⁻¹. Wikstroelide M also potently inhibited HIV-1IIIB induced cytolysis in MT-4 cells, with an EC50 value of 9.60 ng·mL⁻¹. The mechanistic assay showed that wikstroelide M targeted HIV-1 reverse transcriptase and nuclear translocation of integrase through disrupting the interaction between integrase and LEDGF/p75.
CONCLUSION
Wikstroelide M may be a potent HIV-1 and HIV-2 inhibitor, the mechanisms of action may include inhibition of reverse trascriptase activity and inhibition of integrase nuclear translocation through disrupting the interaction between integrase and LEDGF/p75.
Anti-HIV Agents
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pharmacology
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therapeutic use
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Cell Line
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Daphne
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chemistry
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Diterpenes
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pharmacology
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HIV Infections
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drug therapy
;
virology
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HIV Integrase
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metabolism
;
HIV Integrase Inhibitors
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pharmacology
;
therapeutic use
;
HIV Reverse Transcriptase
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antagonists & inhibitors
;
HIV-1
;
drug effects
;
enzymology
;
HIV-2
;
drug effects
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Humans
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Intercellular Signaling Peptides and Proteins
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metabolism
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Phytotherapy
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Plant Extracts
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pharmacology
;
therapeutic use
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Virus Integration
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drug effects
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Virus Replication
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drug effects