Browsing by Author "Samarakoon, K. W."

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Anti-cancer activity of Cinnamomum zeylanicum, Pleiospermium alatum, Scolopia pusilla, Zingiber officinale, Allium sativum, and Allium cepa against breast and cervical cancer cell lines
(Faculty of Science, University of Kelaniya Sri Lanka, 2024) Jayasinghe, M. R.; Jayasundara, N.; Samarakoon, K. W.; Kuruppu, A.I.
Despite advancements in modern medicine, cancer remains a global challenge, with existing treatments being costly and often accompanied by side effects. Developing novel therapies from herbal-based products presents a promising alternative. This study evaluates the anticancer potential of a blend of herbs of a traditional formula: Cinnamomum zeylanicum, Pleiospermium alatum, Scolopia pusilla, Zingiber officinale, Allium sativum, and Allium cepa. The formula was prepared by extracting these herbs individually using methanol, and then combining the extracts in equal proportions. The combined methanolic extract (formula) was subjected to various assays to assess its therapeutic potential. The methanolic extracts of all herbs were tested by the Folin-Ciocalteu's reagent method and the aluminum chloride colorimetric method to estimate the quantity of phenol and flavonoid content, respectively. The 2, 2-diphenyl 1-picrylhydrazyl (DPPH) assay and the human red blood cell (HRBC) membrane stabilization assay assessed the antioxidant and anti-inflammatory effects, respectively. The MTT (3- (4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide) assay and the clonogenic assay were used to examine the cytotoxicity impact in vitro. At the maximum dosage tested (1 mg/mL), C. zeylanicum showed a somewhat higher concentration of total phenols, with a value of 0.478±0.008 mg/mL GAE/g, while the formula with all herbs, represented 0.305 ± 0.007 mg/mL GAE/g. The P. alatum extract had the greatest total flavonoid content, measuring 0.050 ± 0.035 mg/ml QE/g. The formula's methanolic extract showed a TFC of 0.039 ± 0.019 mg/mL QE/g. The single herb S. pusilla extract's IC50 was the lowest among single herbs for antioxidant activity, 0.0632 ± 0.008 mg/mL; in contrast, the formula's methanolic extract showed a lower IC50 0.0421 ± 0.004 mg/mL. The formula yielded an IC50 of 0.0432±0.002 mg/mL, whereas the IC50 value for C. zeylanicum extract showed 0.052±0.014 mg/mL for the anti-inflammatory assay. Z. officinale extract showed an inhibitory effect on the MCF-7 cell line, with an IC50 value of 0.0514 ± 0.012 mg/mL, while the formula showed a value of 0.0362 ± 0.081 mg/mL. The extract of Z. officinale produced growth inhibitory effects against the HeLa cell line with an IC50 value of 0.0840 ± 0.041 mg/mL, whereas the formula showed a more potent IC50 of 0.0273 ± 0.002 mg/mL indicating that the formula was potent against the cancer cells. Vero cells, non-cancerous monkey kidney cells, showed an IC50 of >1 mg/mL for all herb extracts, which demonstrated a low potency. There was a decrease in the colony-forming capacity with the formula. A remarkable inhibition of colony formation was observed with the formula’s methanolic extract with a survival fraction (SF) of 57.14% against HeLa while it showed a SF of 56.04% for MCF7 cells. This summarizes the information regarding the herbs and their formula, with a special focus on anticancer properties that serve as a foundation for future research and development of this formula in the field of cancer.
Caspase 3/7 activation during apoptotic cell death of human Rhabdomyosarcoma (RMS) and breast adenocarcinoma (MCF-7) cells induced by different fractions of Chnoospora minima
(Faculty of Science, University of Kelaniya, Sri Lanka., 2021) Gunathilaka, M. D. T. L.; Samarakoon, K. W.; Ranasinghe, P.; Peiris, L. C. D.
Marine seaweeds are a rich source of bioactive metabolites that can be used as a source for the development of anti-cancer drugs. Apoptosis is a form of programmed cell death involved with the elimination of unwanted cells from the body. Among different mechanisms of apoptosis, caspases are a family of protease enzymes playing an essential role in apoptosis. Therefore, the present study was aimed to determine the caspase 3/7 activation in human rhabdomyosarcoma (RMS) and breast adenocarcinoma (MCF-7) cells following treatment with hexane and chloroform fractions of the seaweed species Chnoospora minima. The apo-one homogenous caspase 3/7 activity of treated cells was evaluated according to the manufacturer`s instructions (G7790, Promega, USA). Polysaccharide depleted polyphenol-rich methanol extract was sequentially partitioned with hexane, chloroform, and ethyl acetate to determine the cytotoxic activity. Based on the results, hexane and chloroform fractions of C.minima were selected to determine the caspase 3/7 activation of human RMS and MCF-7 cells. The caspase 3/7 activation was quantified by relative flurescence unit (RFU). The chloroform fraction (RFU4 hrs:3932.9) of C.minima showed prominent activation of caspase 3/7 in RMS cells after 4 h of caspase treatment more than the hexane fraction (RFU4 hrs:2556.6) compared to the standard Staurosporine (RFU4 hrs:3417.5) and cycloheximide (RFU4 hrs:2950.5). In contrast, hexane (RFU3 hrs:1496.9) and chloroform (RFU3 hrs:1464.7) fractions treated MCF-7 cells showed low caspase 3/7 activation, and the highest activity was observed after 3 h of caspase treatment. Hence, it can be concluded that the hexane and chloroform fractions of C.minima induce apoptosis in RMS cells more prominently via the caspase 3/7 pathway compared to the MCF-7 cells. Therefore, further studies should be conducted to confirm the activity of caspase 3 and 7 via gene expression analysis.
Understanding drug likeness of novel compound Fucoxanthin derivative isolated from Chnoospora minima
(Faculty of Science, University of Kelaniya Sri Lanka, 2022) Bandaranayake, U.; Samarakoon, K. W.; Ranasinghe, P.; Peiris, L. D. C.
Non-communicable diseases pose an ever-increasing burden to many worldwide, regardless of gender, age, and nationality. In Sri Lanka, type 2 diabetes is prevalent due to the rapid transition of lifestyles, unhealthy dietary changes, and population demographic changes, which could be why the epidemic is escalating in South Asia. Advances in chemical biology have expanded the understanding of the marine environment as a diverse source of important bioactive compounds. These organisms include animals, microorganisms, and, most importantly, marine algae. Previously, a derivative of Fucoxanthin has been successfully isolated from Marine algae Chnoospora minima, and this study characterizes and studies the compound computationally using in-silico solubility, toxicity, and molecular docking studies. The compound was drawn using ChemDraw (version 12.0) and was energy minimized using Chem3Dpro (version 12.0). The energy-minimized structure was used in further analysis. Toxicity and aqueous solubility predictions were conducted using OSIRIS Property Explorer. The solubility was expressed in mol/l, and the value of the corresponding log was -6.17. Nearly 80% of the drugs on the market have an estimated logS value greater than -4. Therefore, the compound displays a lower solubility. The partition coefficient of the compound was predicted in cLogP value, which was -8.91. Chemical Property prediction such as half-life, AMES toxicity, and degradation patterns of the compound was made using the EPIsuite (version 4.11). The compound was deemed nontoxic and had a half-life of 14.603 Min. The compound was classified as recalcitrant in terms of bio-degradation. Auto dock tools (version 4.2.0) were used for molecular docking studies against human pancreatic α-amylase and α-glucosidase proteins, and the binding energies were -6.56 kcal/mol and -4.83 kcal/mol, respectively. The ligand formed hydrogen bonds with the protein residue of α-amylase, Arg92, and Asn250, and the residues Pro690, Arg696, and Leu811 were essential for α-glucosidase and ligand binding. The docking procedure was repeated for both proteins with the known drug Acarbose as the ligand. For α-amylase protein binding energy was -4.08 kcal/mol and for α-glucosidase protein binding energy was -3.40 kcal/mol. The data suggest that the novel ligand shows high suitability as a drug based on docking studies. The data suggest that the novel compound could be developed into a herbal supplement against type- 2 Diabetes Mellitus.