Anti-cancer activity of pyridoxal phosphate and metformin combination in human pancreatic cancer cells.

Background: Pancreatic cancer is the foremost cause of cancer-related deaths in many developed countries with a poor prognosis. With advanced disease conditions chemotherapy, surgery followed by radiation is the regimen to prolong the survival. But a complete cure is questionable. Metformin is the first-line drug used for the treatment of type 2 diabetes in the world. Aim: The study aims to assess the anti-cancer activity of metformin with the combination of micronutrient pyridoxal phosphate (PLP) in the human pancreatic cancer cell line (PANC-1). Methods: Panc1 cells were maintained in vitro cell culture conditions. The IC50 concentrations of metformin and PLP were estimated and selected by using MTT assay. Morphological changes upon treatments were observed under microscope. Distribution of cells pattern was observed with propidium iodide dye in cell cycle assay. Different phases of cell distribution were studied with apoptosis assay. Results: More morphological changes were observed with PLP followed metformin. MTT assay revelled the IC50 concentrations of metformin and PLP were 20.95 ± 0.98 mM and 5.70 ± 0.07 mM. The cell cycle assay revealed that the percentage of cells was arrested in different phases with the treatments. Apoptosis assay revelled metformin increased necrosis population to 9.9%, whereas PLP has enhanced to 14.2% apoptosis. Tumour suppressor protein p53 levels had increased to 24.8% with PLP and 3.5% with metformin. Conclusion: In conclusion, PLP has significantly induced cell cycle arrest, apoptosis and enhanced p53 protein expression but a combination of PLP with metformin drug has not synergised anti-cancer activity in human PANC1 cells.

UGT1A1 and UGT1A3 activity and inhibition in human liver and intestinal microsomes and a recombinant UGT system under similar assay conditions using selective substrates and inhibitors.

In vitro enzyme kinetics and inhibition data was compared for UGT1A1 and UGT1A3 isoforms under similar assay conditions using human liver microsomes (HLM), human intestinal microsomes (HIM) and recombinant UGT (rUGT) enzyme systems.UGT1A1 catalysed ß-estradiol 3-ß-D-glucuronide formation showed allosteric sigmoidal kinetics in all enzyme systems; while UGT1A3 catalysed CDCA 24-acyl-ß-D-glucuronide formation exhibited Michaelis-Menten kinetics in HLM, substrate inhibition kinetics in HIM and rUGT systems. Corresponding Km or S50 concentrations of ß-estradiol and CDCA were employed in the respective UGT inhibition studies.Atazanavir inhibited the production of ß-estradiol 3-ß-D-glucuronide with IC50 values of 0.54 µM and 0.16 µM in HLM and rUGT1A1, respectively. But its inhibition potential was not observed in HIM, indicating potential cross-talk with other high-affinity intestinal UGT isozymes. On the other hand, zafirlukast, a pan UGT inhibitor, exhibited moderate inhibition in HIM with an IC50 value of 16.70 µM. Lithocholic acid, inhibited the production of CDCA 24-acyl-ß-D-glucuronide with IC50 values of 1.68, 1.84, and 12.42 µM in HLM, rUGT1A3, and HIM, respectively.These results indicated that HLM, HIM, and rUGTs may be used as complementary in vitro systems to evaluate hepatic and intestinal UGT mediated DDIs at the screening stage.

Chumacin-1 and Chumacin-2 from Pseudomonas aeruginosa strain CGK-KS-1 as novel quorum sensing signaling inhibitors for biocontrol of bacterial blight of rice.

The in vitro inhibition of quorum sensing signal, xanthan gum secretion, biofilm formation in different Xanthomonas pathovars and biological control of bacterial blight of rice by the two bioactive extrolites produced by Pseudomonas aeruginosa strain CGK-KS-1 were explored. These extrolites were extracted from Diaion HP-20 resin with methanol and purified by preparative-thin layer chromatography. Further, spectroscopic structural elucidation revealed the tentative identity of these extrolites to be (R,3E,5E,9Z,11E)-13-((3S,5R)-5-acetyl-2,6-dimethylheptan-3-yl)-10-hydroxy-4-methyl-1,8-diazabicyclo[9.3.1]pentadeca-3,5,9,11(15),13-pentaen-2-one and (R,3E,5E,8E,11E)-13-((3S,5R)-5-acetyl-2,6-dimethylheptan-3-yl)-4-methyl-1,8-diazabicyclo[9.3.1]pentadeca-3,5,8,11(15),13-pentaene-2,10-dione, named as Chumacin-1 and Chumacin-2, respectively. Antimicrobial assay showed Chumacin-1 and Chumacin-2 exhibited a strong in vitro growth inhibition against various Xanthomonas pathovars. Quorum sensing overlay assay using a reporter strain Chromobacterium violaceum strain CV026 showed that Chumacin-1 and Chumacin-2 inhibited quorum sensing signaling. The mechanistic studies revealed that these extrolites inhibited the production of quorum sensing signaling factor, cis-11-methyl-2-dodecenoic acid; suppressed the xanthan gum secretion and also inhibited the biofilms formed by various Xanthomonas pathovars. Both Chumacin-1 and Chumacin-2 showed ROS generation in the test Xanthomonas strains, resulting in in vitro cell membrane damage was revealed through CSLM and FE-SEM micrographs. Further, greenhouse experiments using Samba Mashuri (BPT-5204) revealed that seed treatment with Chumacin-1 and Chumacin-2 along with foliar spray groups showed up to ˜80% reduction in bacterial blight disease in rice. To the best of our knowledge, this is the first report on new quorum sensing inhibitors, Chumacin-1 and Chumacin-2 produced by Pseudomonas aeruginosa strain CGK-KS-1 exhibiting DSF inhibition activity in Xanthomonas oryzae pv. oryzae.

Novel dihydropyrimidine derivatives as potential HDAC inhibitors: in silico study.

Dihydropyrimidine derivatives possess many biological activities due to presence of pyrimidine ring structure in various nucleic acids, vitamins, coenzymes, uric acid and their derivatives. They have possessed broad spectrum actions like antibacterial, antifungal, antiviral, anticancer and antihypertensive etc. Before synthesis of compounds, it is good to predict biological activity using in silico methods. Here, we have selected some of N (3a-f) and O (4a-f) mannich bases of dihydro pyrimidine derivatives emphasized on histone deacetylase 4 (HDAC-4) inhibitions activity. We have used the different software tools like Lipinski's rule of five; pass online; osiris property explorer and docking studies to predict anti cancer activity. All the selected compounds exhibited potential drug like molecule with anti cancer activity. Among all compound the substitution with methoxy group (3c) exhibited more drugs like property and substation with hydrogens (4a) showed high anti neoplastic activity; whereas substitution with dichloro groups (4e) showed more drug docking scores. These were compared with standard drugs tamoxifen and 5-flourouracil. The approach of predicting anticancer activity using in silico method may be more useful to select and synthesis novel compounds in research as well as in industry.