Synergistic anti-hepatitis C virus activity of Ruta angustifolia extract with NS3 protein inhibitor

Background Medicinal plants are known to perform many pharmacological actions due to their chemical metabolites, which include antiviral effects. Previously, the extract of Ruta angustifolia was shown to have potential anti-hepatitis C virus (HCV) activity without any cytotoxicity, with a 50% inhibitory concentration of 3.0 μg/mL and a 50% cytotoxicity concentration of >100 μg/mL. Furthermore, the combination of medicinal plants and current anti-HCV agents, such as a direct-acting antiviral agent, was shown to potentiate their overall effectiveness. In the course of this study, the ethanolic extract of R. angustifolia was evaluated for its anti-HCV effects; specifically, the mechanism of action on HCV NS3 and NS5A protease was investigated. Methods Analysis of the use of this extract in conjunction with current NS3 inhibitor drugs, simeprevir (SMV) and telaprevir (TVR), was performed. Anti-HCV activity was performed by in vitro culture of hepatocyte cells. The cells were infected and treated with various concentrations of the sample. HCV inhibition was calculated and CompuSyn software analysis was used to determine the synergistic effect of the combination. Results Results demonstrated that R. angustifolia extract inhibited the post-entry step and decreased the protein levels of HCV NS3 and NS5A. The combination of extract and SMV and TVR mediated a synergistic effect. Conclusions These findings suggest that combining R. angustifolia extract with current anti-HCV drugs should be considered when developing alternative and complementary anti-HCV medicines

The Role of Polysaccharide Krestin From Coriolus versicolor Mushroom on Immunoglobulin Isotype of Mice Which Infected by Mycobacterium tuberculosis

This research was aimed to determine the role of polysaccharide krestin (PSK) with different timing on levels and types of mice immunoglobulin (Ig) isotype which infected by Mycobacterium tuberculosis. This research used 30 adult female mice of Mus musculus strain, polysaccharide krestin was isolated from Coriolus versicolor mushroom, and for infection used Mycobacterium tuberculosis H37Rv (ATCC 27294 T) strain.Provision of polysaccharide krestin was done over 7 consecutive days via gavage. Mycobacterium tuberculosis infection was done 2 times with an interval of 1 week via intraperitoneal. Immunoglobulin isotype serums were analyzed using the ELISA test and the results were analyzed descriptively through the color reaction and OD values. The result showed the highest levels of immunoglobulin was found in the provision of PSK before and after Mycobacterium tuberculosis infection with total 6.280 of OD Ig isotype. Immunoglobulin isotype dominant was IgM with lambda light chain. The conclusion of this research was PSK increased mice Ig isotype levels at the time of provision before, after or before and after infection Mycobacteriummtuberculosis. Ig isotype which wasm formed i.e. IgM, IgA, IgG2b, IgG3, IgG2a, IgG1 with kappa and lambda light chain

Anticancer activity of okra raw polysaccharides extracts against human liver cancer cells

Purpose: To examine raw polysaccharides extract of okra (Abelmoschus esculentus L.) as an anticancer agent against human liver cancer cells. Methods: Okra raw polysaccharide extract (ORPE) was obtained by ethanol extraction from the raw fruit. Huh7it cells were grown in DMEM (Dulbecco's Modified Eagle Medium) and cultured for 24 h prior to treatment with the extract. The cell culture was divided into 3 groups, viz, negative control group (KN), positive control group (KP, treated with 10 μg/mL doxorubicin), and ORPE (P) group. ORPE group was divided into 5 subgroups based on the dose used for treatment, viz, 50 (P1), 100 (P2), 200 (P3), 400 (P4), and 600 µg/mL (P5). Huh7it cell proliferation was measured by MTT assay. while measurement of Huh7it cell apoptosis, necrosis, and cell cycle analysis were carried out using Annexin V FITC-PI antibody test and flow cytometry. Results: ORPE significantly inhibited Huh7it cell proliferation and induced apoptosis. ORPE treatment with 600 µg/mL extract caused 5.82 % late cell apoptosis and 5.62 % early apoptosis. Cell cycle arrest occurred during G0/G1 phase. Conclusion: ORPE is a potential anticancer agent against human liver cancer cells due to its ability to induce apoptosis of huh7it cells by promoting cell cycle arrest during G0/G1 phase

Anti-viral activity of Phyllanthus niruri against hepatitis C virus

Hepatitis C virus (HCV) infection is a global problem that causes liver disease and hepatocellular carcinoma. Although the current standard treatment provided a significant improvement on response rate with sustain virology response more than 90%, however, the high cost was remaining limited access to this therapy, resistance emergence and serious side effects which provide the necessities to find the new anti-HCV agents. The current study, we evaluated the ethanol extract of Phyllanthus niruri for its anti-HCV activities. Anti-HCV activity was determined by in vitro culture cells of Huh 7it. Anti-HCV activity of P. niruri extract revealed strong inhibition against HCV with IC50 values of 4.14 µg/mL and yield stronger activity in the entry step of the HCV life cycle. Moreover, the P. niruri extract enhanced anti-HCV activity of simeprevir (NS3 protease inhibitor) with increase the activity up to 4-fold compared to a single treatment of simeprevir. Docking analysis was performed to predict the interaction phyllanthin and hypophyllantin, known compounds of P. niruri against HCV receptor. Both of phyllantin and hypophyllantin were mediated a strong interaction with 4GAG, a protein that involved in entry step of HCV. These results suggested that the ethanol extract of P. niruri may be good candidates for the development of anti-HCV drugs

AntiHepatitis C Virus Activity of Alectryon serratus Leaves Extract

Hepatitis C Virus (HCV) has infected approximately 2-3% (130-170 million) of the world's population. No vaccine is available to prevent HCV infection. Investigation of anti-HCV agent is thus deemed necessary. Various plants have been explored for their anti-HCV activity. A. serratus is a member of Sapindaceae family, which fruit and seed were traditionally used as insecticide. Anti-HCV activity tested on A.serratus leaves extract has been done. The result showed that leaves extract exhibited anti-HCV with IC50 value of 14.9 μg/ml and 9.8 μg/ml against HCV J6/JFH1 and JFH1a, respectively. The cytotoxicity assay results showed that A.serratus leaves extract was not toxic and has CC50 >100 μg/ml. Mode of action experiment results suggested that A.serratus extract inhibited HCV at the post-entry step. Further fractionation of leaves extract by open column chromatography resulted in 4 fractions. Only Fraction 1 (AP-5F.1) exhibited anti-HCV with IC50 value of 1.2 μg/ml against HCV JFH1a. Separation of AP-5F.1 by open column chromatography resulted in 15 fractions. Fraction number 13 (AP-5F.1.13) exhibited anti-HCV with IC50 value of 0.43 μg/ml against HCV JFH1a. Separation of AP-5F.1.13 by semi preparative-HPLC resulted in isolate identified by TLC and LC-MS method as chlorophyll derivate. There was a possibility that chlorophyll derivate has participated in performing the anti-HCV activity of fractions and extract besides the other compounds contained. In this study, we concluded that A. serratus leaves extract, AP-5F.1, and AP-5F.1.13 exhibited anti-HCV activity against JFH1a virus

ANTIHEPATITIS C VIRUS ACTIVITY OF INDONESIAN MAHOGANY (TOONA SURENI)

 Objective: Toona sureni (Indonesian mahogany) is a member of Meliaceae family and locally known as suren. Previous study reported that T. sureni leaves extract exhibited antiviral activity with 50% inhibitory concentration (IC50) value of 13.9 ± 1.6 μg/ml against hepatitis C virus (HCV) J6/JFH1. Cytotoxicity analysis of T. sureni leaves extract did not reveal any cytotoxicity effect; therefore, further study was taken to investigate the active substances from the extract.Methods: Bioassay-guided isolation of anti-HCV was conducted using Huh-7.5 cells infected with HCV J6/JFH1 in the presence of extracts, fractions, or compounds from the plant.Results: Ethyl acetate fraction (Fr E) exhibited high anti-HCV activity with IC50 value of 1.7 μg/ml. Further, separation of Fr E by open column chromatography resulted in nine sub-fractions (sub-Fr E1-E9). Sub-Fr E3 and E4 have IC50 value of 29.90 μg/ml and 7.68 μg/ml, respectively. Polyphenols compounds have been isolated from sub-Fr E3 and E4. The structures have been determined to be ethyl gallate (1), methyl gallate (2), catechin (3), gallic acid (4), and quercetin 3-O-rhamnoside (5). Among the isolated compounds, gallic acid showed to possess strong anti-HCV activity with IC50 value of 15.9 μg/ml.Conclusion: T. sureni and its isolated compound, gallic acid, may be good candidates to develop for alternative and/or complementary agents of anti-HCV infection

Antiviral Activities of Indonesian Medicinal Plants in the East Java Region Against Hepatitis C Virus

Background Hepatitis C virus (HCV) is a major cause of liver disease and a potential cause of substantial morbidity and mortality worldwide. The overall prevalence of HCV infection is 2%, representing 120 million people worldwide. Current standard treatment using pegylated interferon and ribavirin is effective in only 50% of the patients infected with HCV genotype 1, and is associated with significant side effects. Therefore, it is still of importance to develop new drugs for treatment of HCV. Antiviral substances obtained from natural products, including medicinal plants, are potentially good targets to study. In this study, we evaluated Indonesian medicinal plants for their anti-HCV activities. Methods Ethanol extracts of 21 samples derived from 17 species of medicinal plants explored in the East Java region were tested. Anti-HCV activities were determined by a cell culture method using Huh7.5 cells and HCV strains of 9 different genotypes (1a to 7a, 1b and 2b). Results Four of the 21 samples tested showed antiviral activities against HCV: Toona sureni leaves (TSL) with 50% inhibitory concentrations (IC50) of 13.9 and 2.0 μg/ml against the HCV J6/JFH1-P47 and -P1 strains, respectively, Melicope latifolia leaves (MLL) with IC50 of 3.5 and 2.1 μg/ml, respectively, Melanolepis multiglandulosa stem (MMS) with IC50 of 17.1 and 6.2 μg/ml, respectively, and Ficus fistulosa leaves (FFL) with IC50 of 15.0 and 5.7 μg/ml, respectively. Time-of-addition experiments revealed that TSL and MLL inhibited both at the entry and post-entry steps while MMS and FFL principally at the entry step. TSL and MLL inhibited all of 11 HCV strains of all the genotypes tested to the same extent. On the other hand, FFL showed significantly weaker inhibitory activities against the HCV genotype 1a strain, and MMS against the HCV strains of genotypes 2b and 7a to a lesser extent, compared to the other HCV genotypes. Conclusions Ethanol extracts of TSL, MLL, MMS and FFL showed antiviral activities against all the HCV genotypes tested with the exception that some genotype(s) showed significant resistance to FFL and to MMS to a lesser extent. These plant extracts may be good candidates for the development of anti-HCV drug

Antimalarial dihydrochalcone isolated from Artocarpus sericicarpus Jarret leaves and in silico investigation against falcipain-2 protein

Aims: To investigate the potential antimalarial plant belonging to the Artocarpus genus, Artocarpus sericicarpus, and isolate the antimalarial active compound from leaves extract. Methods: The isolation of the antimalarial compound was based on bioassay-guided isolation. The compound isolation was conducted by chromatography and identified using spectroscopic techniques, including 1H, 13C, and 2D NMR. The compound was tested for its antimalarial activity against Plasmodium falciparum and evaluated for cytotoxicity using several cell lines, including Huh7, HepG2, BHK-21, and Vero cells. In silico investigation of the compound against falcipain-2 protein was conducted as well. Results: Fractionation and purification of dichloromethane leaves extract led to the isolation of a dihydrochalcone compound identified as 1-(2,4-dihydroxyphenyl)-3-[4-hydroxy-3-(3-methylbut-2-en-1-yl)phenyl]propan-1-one. The dihydrochalcone compound exhibited antimalarial activity with an IC50 value of 34.80 µM. Cytotoxicity test revealed CC50 values of the compound were more than 20 µg/mL, and the selectivity indexes (SI) were 4.50, 5.52, 3.02, and 6.68 on Huh7, HepG2, BHK-21, and Vero cells, respectively. The CC50 and SI indicated the nontoxic criteria of the compound. In silico investigation showed that the compound could bind to the falcipain-2 active sites. Conclusions: The dihydrochalcone compound identified as 1-(2,4-dihydroxyphenyl)-3-[4-hydroxy-3-(3-methylbut-2-en-1-yl)phenyl]propan-1-one was isolated from Artocarpus sericicarpus leaves extract showed antimalarial activity and the ability to bind to the falcipain-2 active sites on in silico investigation

Antiviral activity of the dichloromethane extracts from Artocarpus heterophyllus leaves against hepatitis C virus.

Objective: To determine anti-viral activities of three Artocarpus species: Artocarpus altilis, Artocarpus camansi, and Artocarpus heterophyllus (A. heterophyllus) against Hepatitis C Virus (HCV). Methods: Antiviral activities of the crude extracts were examined by cell culture methodusing Huh7it-1 cells and HCV genotype 2a strain JFH1. The mode of action for anti-HCVactivities was determined by time-of-addition experiments. The effect on HCV RNAreplication and HCV accumulation in cells were analyzed by quantitative reversetranscription-PCR and western blotting, respectively. Results: The dichloromethane (DCM) extract of A.heterophyllus exhibited strong anti HCV activity with an inhibitory concentration (IC) value of (1.5 ± 0.6) mg/mL without obvious toxicity. The DCM extracts from Artocarpus altilis and Artocarpus camansi showed moderate anti-HCV activities with IC 50 50 values being (6.5 ± 0.3) mg/mL and (9.7 ± 1.1)mg/mL, respectively. A time-of-addition studies showed that DCM extractfrom A. heterophyllus inhibited viral entry process though a direct virucidal activity and targeting host cells. HCV RNA replication and HCV protein expression were slightly reduced by the DCM treatment at high concentration. Conclusions: The DCM extract from A. heterophyllus is a good candidate to develop an antiviral agent to prevent HCV grant reinfection following liver transplantation

THE ROLE OF POLYSACCHARIDE KRESTIN FROM Coriolus versicolor MUSHROOM ON IMMUNOGLOBULIN ISOTYPE OF MICE WHICH INFECTED BY Mycobacterium tuberculosis

This research was aimed to determine the role of polysaccharide krestin (PSK) with different timing on levels and types of mice immunoglobulin (Ig) isotype which infected by Mycobacterium tuberculosis. This research used 30 adult female mice of Mus musculus strain, polysaccharide krestin was isolated from Coriolus versicolor mushroom, and for infection used Mycobacterium tuberculosis H37Rv (ATCC 27294 T) strain. Provision of polysaccharide krestin was done over 7 consecutive days via gavage. Mycobacterium tuberculosis infection was done 2 times with an interval of 1 week via intraperitoneal. Immunoglobulin isotype serums were analyzed using the ELISA test and the results were analyzed descriptively through the color reaction and OD values. The result showed the highest levels of immunoglobulin was found in the provision of PSK before and after Mycobacterium tuberculosis infection with total 6.280 of OD Ig isotype. Immunoglobulin isotype dominant was IgM with lambda light chain. The conclusion of this research was PSK increased mice Ig isotype levels at the time of provision before, after or before and after infection Mycobacterium tuberculosis. Ig isotype which was formed i.e. IgM, IgA, IgG2b, IgG3, IgG2a, IgG1 with kappa and lambda light chain