Exploring the Traditional Chinese Medicine (TCM) database chemical space to target I7L protease from monkeypox virus using molecular screening and simulation approaches.

In the current study, we used molecular screening and simulation approaches to target I7L protease from monkeypox virus (mpox) from the Traditional Chinese Medicines (TCM) database. Using molecular screening, only four hits TCM27763, TCM33057, TCM34450 and TCM31564 demonstrated better pharmacological potential than TTP6171 (control). Binding of these molecules targeted Trp168, Asn171, Arg196, Cys237, Ser240, Trp242, Glu325, Ser326, and Cys328 residues and may affect the function of I7L protease in in vitro assay. Moreover, molecular simulation revealed stable dynamics, tighter structural packing and less flexible behaviour for all the complexes. We further reported that the average hydrogen bonds in TCM27763, TCM33057, TCM34450 and TCM31564I7L complexes remained higher than the control drug. Finally, the BF energy results revealed -62.60 ± 0.65 for the controlI7L complex, for the TCM27763I7L complex -71.92 ± 0.70 kcal/mol, for the TCM33057I7L complex the BF energy was -70.94 ± 0.70 kcal/mol, for the TCM34450I7L the BF energy was -69.94 ± 0.85 kcal/mol while for the TCM31564I7L complex the BF energy was calculated to be -69.16 ± 0.80 kcal/mol. Although, we used stateoftheart computational methods, these are theoretical insights that need further experimental validation.

p.R180C mutation of glycosyltransferase B leads to B subgroup, an in vitro and in silico study.

BACKGROUND AND OBJECTIVES: Dysfunctional glycosyltransferase A or B may lead to incomplete glycosylation of H antigen and atypical ABO blood group with weak A or B phenotypes, posing challenges for blood typing for transfusion. MATERIALS AND METHODS: Serological studies and ABO gene analysis were performed. Flow cytometry was performed on HeLa cells transfected glycosyltransferase B expressing plasmids. Agglutination of transfected cells and total glycosyltransferase B transfer capacity were examined. Molecular dynamics simulations were used to explore possible dynamic conformational changes around the binding pocket. RESULTS: We identified a mutation c.538C>T (p. R180C) of B allele in a Chinese donor and his father with ABw phenotype. In vitro expression study showed that mutation p.R180C, although not affecting expression of glycosyltransferase B, impaired H to B antigen conversion. The in silico analyses found that the residue Arg180 on the internal loop next to the entry of the binding pocket may have its long side chain salt-bridged with the highly flexible C-terminal carboxyl and contribute to the catalysis of H to B antigen conversion. CONCLUSION: The p.R180C mutation impairs the conversion from H to B antigen and leads to weak B phenotype. Dynamic interaction between Arg180 and C-terminal of glycosyltransferase B may stabilize its binding with UDP-galactose and facilitate H/B antigen conversion.

Cleavage mechanism of the H5N1 hemagglutinin by trypsin and furin.

The cleavage property of hemagglutinin (HA) by different proteases was the prime determinant for influenza A virus pathogenicity. In order to understand the cleavage mechanism, molecular modeling tools were utilized to study the coupled model systems of the proteases, i.e., trypsin and furin and peptides of the cleavage sites specific to H5N1 and H1 HAs, which constitute models of HA precursor in complex with cleavage proteases. The peptide segments 'RERRRKKR downward arrow G' and 'SIQSR downward arrow G' from the high pathogenic H5N1 H5 and the low pathogenic H1N1 H1 cleavage sites were docking to the trypsin and furin active pockets, respectively. It was observed through the docking studies that trypsin was able to recognize and cleave both the high pathogenic and low pathogenic hemagglutinin, while furin could only cleave the high pathogenic hemagglutinin. An analysis of binding energies indicated that furin got most of its selectivity due to the interactions with P(1), P(4), and P(6), while having less interaction with P(2) and little interactions with P(3), P(5), P(7), and P(8). Some mutations of H5N1 H5 cleavage sequence fitted less well into furin and would reduce high pathogenicity of the virus. These findings hint that we should focus at the subsites P(1), P(4), and P(6) for developing drugs against H5N1 viruses.

Virtual screening for finding natural inhibitor against cathepsin-L for SARS therapy.

Recently Simmons et al. reported a new mechanism for SARS virus entry into target cells, where MDL28170 was identified as an efficient inhibitor of CTSL-meditated substrate cleavage with IC(50) of 2.5 nmol/l. Based on the molecule fingerprint searching method, 11 natural molecules were found in the Traditional Chinese Medicines Database (TCMD). Molecular simulation indicates that the MOL376 (a compound derived from a Chinese medicine herb with the therapeutic efficacy on the human body such as relieving cough, removing the phlegm, and relieving asthma) has not only the highest binding energy with the receptor but also the good match in geometric conformation. It was observed through docking studies that the van der Waals interactions made substantial contributions to the affinity, and that the receptor active pocket was too large for MDL21870 but more suitable for MOL736. Accordingly, MOL736 might possibly become a promising lead compound for CTSL inhibition for SARS therapy.

Anti-SARS drug screening by molecular docking.

Starting from a collection of 1386 druggable compounds obtained from the 3D pharmacophore search, we performed a similarity search to narrow down the scope of docking studies. The template molecule is KZ7088 (Chou et al., 2003, Biochem Biophys Res Commun 308: 148-151). The MDL MACCS keys were used to fingerprint the molecules. The Tanimoto coefficient is taken as the metric to compare fingerprints. If the similarity threshold was 0.8, a set of 50 unique hits and 103 conformers were retrieved as a result of similarity search. The AutoDock 3.011 was used to carry out molecular docking of 50 ligands to their macromolecular protein receptors. Three compounds, i.e., C(28)H(34)O(4)N(7)Cl, C(21)H(36)O(5)N(6), and C(21)H(36)O(5)N(6), were found that may be promising candidates for further investigation. The main feature shared by these three potential inhibitors as well as the information of the involved side chains of SARS Cov Mpro may provide useful insights for the development of potent inhibitors against SARS enzyme.

[Effect of yiqi-yangyin prescription on bile composition of cholelithiasis patients].

There were 41 postoperative patients of cholelithiasis with manifestation of Qi-Yin deficiency, had treated with Yiqi Yangyin Prescription. The regimens was 2 weeks as a course, thereafter bile specimens were taken through T tube drainage, and biochemical analysis were performed in order to determine the composition in consequence of the comparison of the treated patients with the control groups. There were 19 patients of bile pigment stone in whom 10 treated with the prescription and 9 were served as control. The results showed that the total bilirubin, unconjugated bilirubin, the concentration of calcium ion and the activity of beta-glucuronidase were markedly decreased as compared to those in the control group (P less than 0.05), the concentration of bile acid markedly increased than that in the control group (P less than 0.05). There were 22 patients of cholesterol stone, 11 treated with the prescription and 11 were served as control group. The results were the same as in bile pigment stone group except decreased in conjugated bilirubin (P less than 0.05), and the unconjugated bilirubin were remained unchanged (P greater than 0.05) as compared to the control group. The ratio of unconjugated bilirubin to total bilirubin and the ratio of bile acid to bilirubin were markedly decreased as compared to those in the control group. The above observation showed promptly that Yiqi-Yangyin Prescription gave a promise influence on lithogenic bile of cholelithiasitic patients, and also investigated the mechanism of the prescription used for deficiency patients with biliary troubles.