Atovaquone and Berberine Chloride Reduce SARS-CoV-2 Replication In Vitro

Epidemic RNA viruses seem to arise year after year leading to countless infections and devastating disease. SARS-CoV-2 is the most recent of these viruses, but there will undoubtedly be more to come. While effective SARS-CoV-2 vaccines are being deployed, one approach that is still missing is effective antivirals that can be used at the onset of infections and therefore prevent pandemics. Here, we screened FDA-approved compounds against SARS-CoV-2. We found that atovaquone, a pyrimidine biosynthesis inhibitor, is able to reduce SARS-CoV-2 infection in human lung cells. In addition, we found that berberine chloride, a plant-based compound used in holistic medicine, was able to inhibit SARS-CoV-2 infection in cells through direct interaction with the virion. Taken together, these studies highlight potential avenues of antiviral development to block emerging viruses. Such proactive approaches, conducted well before the next pandemic, will be essential to have drugs ready for when the next emerging virus hits.

Protective effect of berberine chloride against cisplatin-induced ototoxicity

Background Cisplatin (CP) is an effective anticancer drug broadly used for various types of cancers, but it has shown ototoxicity that results from oxidative stress. Berberine has been reported for its anti-oxidative stress suggesting its therapeutic potential for many diseases such as colitis, diabetes, and vascular dementia. Objective Organ of Corti of postnatal day 3 mouse cochlear explants were used to compare hair cells after the treatment with cisplatin alone or with berberine chloride (BC) followed by CP. Methods We investigated the potential of the anti-oxidative effect of BC against the cisplatin-induced ototoxicity. We observed a reduced aberrant bundle of stereocilia in hair cells in CP with BC pre-treated group. Caspase-3 immunofluorescence and TUNEL assay supported the hypothesis that BC attenuates the apoptotic signals induced by CP. Reactive oxygen species level in the mitochondria were investigated by MitoSOX Red staining and the mitochondrial membrane potentials were compared by JC-1 assay. Results BC decreased ROS generation with preserved mitochondrial membrane potentials in mitochondria as well as reduced DNA fragmentation in hair cells. In summary, our data indicate that BC might act as antioxidant against CP by reducing the stress in mitochondria resulting in cell survival. Conclusion Our result suggests the therapeutic potential of BC for prevention of the detrimental effect of CP-induced ototoxicity.

Identification and estimation of bioactive constituents Negundoside, Berberine chloride, and Marmelosin by HPLC and HPTLC for development of quality control protocols for Ayurvedic medicated oil formulation

Background Anu Taila is an ancient medicated oil Ayurvedic preparation that is commonly used for nasya karma. It contains more than 25 herbs and goat milk as per the Ayurvedic Formulary of India (AFI). It strengthens the neck, shoulder, and chest muscles and improves the capacity of sense organs. It delays the aging process and reduces hair fall. Recent studies showed that it is also useful in COVID-19. In the current study, an attempt to develop quality control protocols and evaluate the standardization parameters like refractive index, iodine value, saponification value, peroxide value, acid value, rancidity, HPTLC fingerprint profile along with major bioactive compound and quantification of Berberine chloride, Negundoside, and Marmelosin by HPLC. Establishing quality protocol and standard parameters like physicochemical parameters and estimation of bioactive compounds of this preparation is significant for quality control. Results In this study, HPTLC identifies bioactive chemical compounds like Berberine chloride, Marmelosin, Negundoside, glycyrrhizin, and para hydroxybenzoic acid (PHBA), Lupeol, Embelin, and Solasodine, which were present in the Anu Taila formulation. HPLC was used to estimate the bioactive marker compounds Negundoside, Berberine chloride, and Marmelosin were present in the Anu Taila formulation. The quantitative evaluation of Berberine chloride (0.0013%), Marmelosin (0.0366%), Negundoside (0.0086%) is present in Anu Taila formulation. Conclusion The study reveals that sufficient quality control parameters were followed during the preparation of the formulation. Physicochemical analysis was carried out as per the guidelines of Ayurvedic Pharmacopeia of India. HPTLC and HPLC profiles generated in this particular study can be considered as a preliminary tool ascertaining the authenticity of Anu Taila.

Berberine Shows Proapoptotic Effect and Potentiation of Cytotoxic Drugs Acitivity in Colorectal Adenocarcinoma Cells

Colorectal cancer, despite advances in treatment and diagnosis, is one of the most common causes of death from cancer. Conventional anticancer treatment is associated with serious side effects, therefore, new strategies are needed to reduce its toxicity. Natural compound berberine has shown antiproliferative activity against various cancer cells including colon. To investigate antiproliferative effect of berberine in human colorectal adenocarcinoma cells and its ability to potentiate the effect of convenient cytostatics taxol, docetaxel, doxorubicin and cisplatin we used MTT assay and flow cytometry was used to detect its proapoptotic effect and effect on cell cycle. In our experiments we observed significant antiproliferative activity of berberine chloride with IC50= 6.7526 μM. Combination of berberine chloride (1μM) with taxol, docetaxel, doxorubicin and carboplatin significantly increased their antiproliferative activity and decreased their IC50 values. Cell cycle analysis revealed increase in sub-G0 /G1 fraction considered as apoptotic and very mild increase in G0/G1 fraction in berberine chloride (5μM) treated cells. proapoptotic effect of berberine chloride (5μM) was confirmed by Annexin V/PI staining. Based on our results, we can assume that berberine has antiproliferative effect and the combination of berberine with cytostatics could be a promising alternative treatment for cancer. Further studies are necesary to investigate the exact mechanisms of action and drug – drug interactions.

ASSESSMENT OF PERMEABILITY BEHAVIOR OF BERBERINE CHLORIDE ACROSS GOAT INTESTINAL MEMBRANE IN PRESENCE OF NATURAL BIOPOTENTIATOR CURCUMIN

The present study investigates the influence of co-administration of different concentrations (2, 6, and 10 mg) of curcumin on goat intestinal permeability of berberine chloride (BBC) using Franz diffusion cell. Data obtained in triplicate from permeability studies were used to calculate percentage cumulative drug release (% CDR), apparent permeability (Papp), flux (J) and enhancement ratio (ER). Co-administration of 6 mg concentration of curcumin with BBC was found to be optimum to enhance the permeability of BBC up to 23.92 ± 0.78 % CDR, over control (8.49 ± 1.45 % CDR). At the optimized concentration of curcumin, permeability characteristics were improved significantly compared to control. The present study reveals the beneficial effect of co-administration of curcumin (6 mg) to promote membrane permeability of BBC which would be expected to improve its bioavailability, thereby therapeutic efficacy. The effect could be attributed to curcumin-mediated inhibition of intestinal efflux pump P-gp, acting as an absorption barrier for BBC.

Co-Delivery of Berberine Chloride and Tariquidar in Nanoliposomes Enhanced Intracellular Berberine Chloride in a Doxorubicin-Resistant K562 Cell Line Due to P-gp Overexpression

The MDR phenomenon has become a major obstacle in the treatment of cancers, and among the strategies to reverse it, the inhibition of P-gp function and expression is essential to increase for effective anticancer drugs. In the present paper, the co-delivery of berberine chloride and tariquidar loaded nanoliposomes was investigated with the aim of enhancing solubility and improving desired effects for the antineoplastic drug and the P-gp inhibitor. Developed nanoliposomes were loaded with the electron-dense enzyme horseradish peroxidase, and analyzed by TEM to investigate their ability to enter in both K562 and K562/DOXO cell lines. Receptor-mediated endocytosis was evidenced for both cell lines. Nanoliposomes were loaded with tariquidar, berberine chloride, or both, maintaining chemical and physical characteristics—i.e., size, homogeneity, and encapsulation efficiency—and high suitability for parenteral administration. Tariquidar was able to reverse the MDR in the K562/DOXO cell line. Tariquidar- and berberine chloride-loaded nanoliposomes showed a significant increase of berberine chloride accumulation in tumor cells, which could be correlated with resensitization of the resistant cells to the antitumor agent. These results suggest that the co-delivery of the P-gp inhibitor, tariquidar, and the cytotoxicity inducer, berberine chloride, looks like a promising approach to overcome the MDR.

The Effect of Resistance Training and Berberine Chloride on the Apoptosis- Related UPR Signaling Pathway in the Hippocampus of Diazinon- Poisoned Rats

ntroduction: Diazinon is one of the most widely used organophosphate pesticides in the world. This toxin enters the body in various ways and induces oxidative stress in various tissues. It has been proved that activation of UPR under oxidative stress is a consistent mechanism for maintaining cell function and survival. Therefore, present study aimed to review the effect of resistance training (RT) and berberine chloride (BC) on the apoptosis- related UPR signaling pathway in the hippocampus of diazinon- poisoned rats. Methods: In this experimental study, 40 male Wistar rats weighing 250 ±50 g were randomly divided into eight groups of five rats including 1) diazinon + 2 mg/kg BC + RT, 2) diazinon + 15 mg/kg BC + RT, 3) diazinon, 4) diazinon + RT, 5) diazinon + 2 mg/kg BC, 6) diazinon + 15 mg/ kg BC, 7) healthy control, 8) sham. The groups were treated for 5 weeks. At the end of the fifth week, ATF-4, ATF-6 and CHOP gene expression in hippocampus tissue were measured by Quantitative Real Time RT-PCR. Results: Diazinon significantly increased the expression of ATF-4, ATF-6, and CHOP in the hippocampus tissue of rats. 15 mg/kg BC with RT significantly decreased these gens, which may indicate a decrease in the rate of apoptosis in the hippocampus. Conclusion: This study showed that RT along with BC has a protective effect against diazinon- induced toxicity in the hippocampus.