Bioinformatic Test and Pharmacokinetic Profile Prediction of Gnetin-C Compound in Melinjo (Gnetum gnemon L.) Seeds Toward Colorectal Cancer

Cancer is one of the unresolved health problems in the world, including Indonesia. One of the most common types of cancer is colorectal cancer. Melinjo (Gnetum gnemon L.) is one of Indonesian local commodities which has many benefits including its potential to be developed as an anticancer agent. Through bioinformatics and molecular docking technology, the aim of this study was to investigate the functions of gnetin-C from melinjo against colorectal cancer. PkCSM database were used to search the ADMET (absorption, distribution, metabolism, elimination and toxicity) properties and stitch-string database were used to identify common genes related to colon cancer. During identification of colon cancer related genes, STAT3 protein showed the highest degree score. Furthermore, molecular docking was carried out to find out the interaction between the STAT3 protein and gnetin c compound found in melinjo seeds. From the docking stage, the pose with the best affinity energy was obtained with a docking score of 1,966 kcal/mol. this shows that the compound gnetin c has the potential to be used as a new anticancer agent from natural ingredients.Keywords: Melinjo, Gnetum gnemon, colon cancer, bioinformatics, molecular docking

Anti-Lung Cancer and Cell Migration Inhibition Properties of Ethyl Acetate Extract of Selaginella doederleinii Towards HTB-183 Cells through In Silico and In Vitro Approach

Continuous research and development to obtain novel anti-lung cancer agents is essential, considering the high prevalence and mortality of the disease. The biflavonoid compounds of Selaginella doederleinii showed significant anticancer activities. This study aims to determine the cytotoxic and cell migration inhibition properties of ethyl acetate extract of Selaginella doederleinii (EAESD) against HTB-183 cells through in silico and in vitro methods. This study started with extraction and then identified biflavonoids in EAESD by HPLC. In vitro analysis was conducted through MTT Assay to observe the cytotoxic properties of EAESD and Wound Scratch Healing Assay to observe its cell migration inhibitory properties. In silico studies to obtain the potential anti-lung cancer compounds and their protein targets were conducted through bioinformatics, combining PASS analysis, Swiss Target Prediction, and STITCH. The obtained compounds and protein targets were analyzed in Molecular Docking to evaluate the binding affinities. The result showed that EAESD contained biflavonoid compounds, exhibited cytotoxic activity with an IC50 value of 190 μg/ml, and inhibited the migration rate of HTB-183 cells. Based on in silico analysis, the three biflavonoids with the highest potential of antilung cancer activity along with their target protein are robustaflavone 7,4-dimethyl ether with EGFR, heveaflavone with ESR1, and 7,4',7'',4'''-tetra-O-methyl-amentoflavonewith TNF. All compounds can bind to each protein target with the docking score -9.2 kcal/mol, -9.5 kcal/mol, and -6.5 kcal/mol, respectively. This study suggested preliminary data regarding the potential of Selaginella doederleinii to inhibit the proliferation and migration of the HTB-183 cell line of lung cancer.Keywords: Selaginella doederleinii, HTB-183, cytotoxicity, cell migration, in silico analysis

Chemopreventive Properties Curcuma heyneana Rhizome Ethanolic Extract on Hepatocellular Carcinoma Cells, JHH-4

Hepatocellular carcinoma is the most common type of liver cancer. Curcuminoids are natural polyphenol compounds abundant in Curcuma heyneana ethanolic extract (CHE) and are known to inhibit breast and cervical cancer cell proliferation. Based on previous research, curcuminoid compounds have been studied to inhibit the growth of the liver cancer cell model, HepG2. This study aims to examine the potential of CHE as a chemopreventive agent in liver cancer using JHH-4 cell as a model. CHE was obtained by maceration method using ethanol which was then identified for its phytochemical profile using thin layer chromatography (TLC). Then TLC results were quantified to calculate the levels of compounds present in the CHE based on spot intensity with ImageJ software. 2,2-Diphenyl-1-picrylhydrazyl (DPPH) antioxidant assay was conducted to determine the radical scavenging activity of CHE. Cytotoxic activity of CHE on JHH-4 liver cancer cells was tested by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Extraction produces a yield of 10.2 %w/w. CHE contains 4.52 %w/w curcuminoid compound consisting of 0.49 %w/w curcumin, 3.21 %w/w demethoxycurcumin, and 0.82% w/w bisdemethoxycurcumin. CHE exhibited antioxidant activity with an IC50 value of 378.96 μg/mL, meanwhile ascorbic acid as a positive control has an IC50 value of 8.49 μg/mL. Cytotoxic activity of CHE on JHH-4 cells is characterized by an IC50 value of 16.62 μg/mL which is classified as having strong cytotoxic activity. This study concluded that CHE has the potential to be developed as a chemopreventive agent in liver cancer.Keywords: liver cancer, hepatocelullar carcinoma, Chemopreventive, antioxidant,Curcuma heyneana

Potential Inhibition of Melaleuca leucadendron L. Compounds Against the NSP5 SARS CoV-2 Protein

COVID-19 is an infectious disease caused by Severe Acute Respiratory Syndrome (SARS-CoV-2), causing a global health emergency as a pandemic disease. The lack of certain drug molecules or treatment strategies to fight this disease makes it worse. Therefore, effective drug molecules are needed to fight COVID-19. Non Structural Protein (NSP5) or called Main Protease (Mpro) of SARS CoV 2, a key component of this viral replication, is considered a key target for anti-COVID-19 drug development. The purpose of this study is to determine whether the compounds in the Melaleuca leucadendron L. plant such as 1,8-cineole, terpene, guaiol, linalol, α-selinenol, β-eudesmol and γ-eudesmol are predicted to have antiviral activity for COVID-19. Interaction of compounds with NSP5 with PDB code 6WNP analyzed using molecular docking with Molegro Virtual Docker. Based on binding affinity, the highest potential as an anti-viral is Terpineol with binding energy (-119.743 kcal/mol). The results of the interaction showed that terpinol has similarities in all three amino acid residues namely Cys 145, Gly 143, and Glu 166 with remdesivir and native ligand. Melaleuca leucadendron L. may represent a potential herbal treatment to act as: COVID-19 NSP5, however these findings must be validated in vitro and in vivo.Keywords: COVID-19, In Silico, NSP5/ 6WNP, Melaleuca leucadendron L

Bioinformatics Analysis of Inhibition Activation SHP-2 by Galangal as Activating Agent of Cancer Immunotherapy

Interleukin 12 (IL-12) is a pro-inflammatory cytokine type 1 that has acted as a potential immunotherapy for cancer. The mechanism of IL-12 increases the activity of cytotoxic T cells and Natural Killer (NK) cells, reverse tumor-induced immunosuppression, prevent angiogenesis, and increases lymphocyte and antigen transport. Galangal is one of the natural ingredients that have biological activity as an anticancer and immunomodulator. In this research, researchers wanted to know the potential of the active compound of galangal to activate IL-12 by inhibiting the IL-12 analog, namely SHP-2. This research uses bioinformatics studies using several databases such as RCSB PDB, ChEMBL, Dr. Duke's Phytochemical and Ethnobotanical, UALCAN, OncoLnc and computational analysis using KNIME and MOE software. The SHP-2 structure used is taken from the RCSB PDB with the code 5EHR. The 10 compounds with the highest predictions of inhibiting SHP2 using KNIME were obtained, then molecular docking was performed using MOE and three compounds that had the potential to inhibit SHP-2 were Kaempferide, Galangin, and RiboflavinKeywords: cancer, computing, galangal, Interleukin 12, SHP-2

Citrus sinensis Peel Extract Synergistically Enhances the Cytotoxic Effect of Chemotherapeutic Agents on HepG2 Cells

Doxorubicin (DOX) and cisplatin (Cis), non-specific chemotherapeutic agents used for hepatocellular carcinoma (HCC), are frequently combined with synthetic or natural agents to enhance their cytotoxic effects. Citrus sinensis peel extract (CPE) serves as a natural source of flavonoids, including sinensetin (SIN), which has the potential to increase the efficacy of DOX and Cis. This study aimed to observe the effect of CPE and SIN one of CPE compounds, in enhancing liver cancer cell susceptibility to doxorubicin and cisplatin. The assays conducted in this study included a phytochemical analysis of CPE using TLC, cell viability assays against HepG2 cells using MTT assay in both single and combination forms, and cell viability assays on Vero cells. The result confirmed the presence of SIN as one of the compounds in CPE. Both CPE and SIN, when used individually, exhibited moderate cytotoxic effects on HepG2 cells with IC50 of 101.09 μg/mL and 83.13 μM, respectively, while showing no cytotoxic effect on Vero cells. Cis demonstrated significant cytotoxicity against HepG2 cells with an IC50 of 7.86 μM. DOX exerted a strong cytotoxic effect on both HepG2 and Vero cells, with the IC50 of 2.52 μM and 13.98 μM. It was observed that CPE was able to synergistically enhance the cytotoxic effects of DOX, and SIN synergistically increased the cytotoxicity of Cis, particularly against HepG2 cells, with CI<1.0.Keywords: CPE, SIN, Cisplatin, Doxorubicin, HCC

Acute Toxicity Test of Ekor Naga (Rhaphidophora pinnata (L.f) Schott) Leaf Extract in Mice and Kidney Histological Examination

Ekor Naga (Rhaphidophora pinnata (L.f) Schott) is a traditional plant used by the community. Previous studies have shown that this plant has pharmacological effects, including as anthelmintics, antioxidant, and anti-inflammatory agent as well as promotes wound healing. Thus, it is necessary to do a toxicity test. The study aimed to determine the effect of the extract on histopathology of the kidney through a toxicity test. The research used an experimental design. The test animals were divided into 5 groups, each of which consisted of 5 test animals: negative control (0.5% Na. CMC), Treatment 1 (Extract of ekor naga leaves at a dose of 200 mg/Kg BW), Treatment 2 (Extract of ekor naga leaves at a dose of 600 mg/Kg BW), Treatment 3 (Extract of ekor naga leaves at a dose of 1800 mg/Kg BW), and Treatment 4 (Extract of Ekor Naga Leaves at a dose of 5400 mg/Kg BW). The parameters observed were the Letal Dose (LD50) and the histophatology of the kidney. Data were analyzed using One Way ANOVA test, followed with Duncan’s test. The results showed no significant difference in the weight of the kidney (p>0.05), and the histophatology of the kidney after 14 days of acute administration of ekor naga leaf extract in all the treatment was normal. The administration of ekor naga leaf extract at therapeutic doses and larger doses of single administration did not have a bad effect on the histology of the kidney.Keywords: Ekor Naga Leaves, Kidney, Histopathology, Acute Toxicity

Cytotoxic and Anti-proliferative Effects of Ethanol Extract of Marine Sponge Stylissa carteri on Colon Cancer HCT-116 Cell Line

Colorectal cancer is one of the most diagnosed cancers in the world. KRAS mutations in colon cancer are being responsible for the progressiveness and resistance of the standard therapeutic available. Marine sponge is one the sources for chemotherapy. Stylissa carteri is a marine sponge that lives in Indonesia and its anti-cancer effects are starting to be explored nowadays. The purpose of this study was to determine the cytotoxicity and anti-proliferative effect of ethanol extract of Stylissa carteri against colon cancer cells with KRAS mutations HCT-116 cells. This was an in vitro study using colon cancer cell line HCT-116 which was tested by 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) and trypan blue assay. The trypan blue assay was performed on control and treated cells at 72 h. The MTT assay was performed on cells which were previously incubated with Stylissa carteri extract. Sponge Stylissa carteri was taken from Pramuka island, Taman Nasional Kepulauan Seribu, Jakarta. The IC50 of ethanol extract of Stylissa carteri is 5 μg/mL (R square 0.9550) in HCT-116 cells. The trypan blue assay showed doubling time of incubated cell for 48 h was 26.10 h for control and 39.69 h for treated cells, respectively. Ethanol extract of Stylissa carteri has cytotoxic and anti-proliferative effects in HCT-116 colon cancer cells with KRAS mutation.Keywords: Colon Cancer, HCT-116, KRAS Mutation, Marine Sponge, Stylissa carteri

Etlingera elatior Compounds as Anticancer Agents of Breast Cancer Through Inhibition of Progesterone Receptor: An In Silico Study

Breast cancer is the leading cause of cancer-related death in women globally. Progesterone receptor (PR) is known as the prime example of receptors amenable to targeted breast cancer drug therapy. Etlingera elatior is an herbal plant that has been renowned to have anticancer effect. This study aimed to identify the potential compounds derived from Etlingera elatior as anticancer agents of PR in breast cancer using molecular docking method. This study used fifteen compounds from Etlingera elatior along with lonaprisan as the comparative drug. The PR was downloaded from RCSB, whereas compounds and lonaprisan were from Pubchem. The drug-likeness test based on Lipinski’s rule of five was conducted using SwissADME. Toxicity analysis using admetSAR 2.0 was used to predict toxicological profile of the compounds. Compounds and lonaprisan were docked on PR using AutoDock tools 1.5.6 and AutoDock Vina 1.1.2. Molecular interactions were visualized by Discovery Studio v16. A total of nine compounds met the criteria as drugs based on drug-likeness and toxicity tests. All nine compounds except caffeic acid and vanillic acid had higher binding affinities on PR compared with lonaprisan. Ergosterol peroxide exhibited the highest binding affinity on PR with values of -9.8 kcal/mol. Moreover, ergosterol peroxide-PR interaction had thirteen hydrophobic bonds and a hydrogen bond with amino acid residues were found in the active site of PR. Most of the compounds found in Etlingera elatior have the potential to be anticancer agents of PR in breast cancer with ergosterol peroxide being the most potential compound. Further in vitro and in vivo research are needed.Keywords: breast cancer, ergosterol peroxide, Etlingera elatior, progesterone receptor, in silico

Cytotoxic Activity of Cambodian Leaves Extract (Plumeria acuminate) on Breast Cancer Cells and COX-2 Targeted Prediction of Its Chemical Contents

Cambodian leaves are suspected to contain stigma sterol which may target Cyclo-Oxygenase-2 (COX-2) or Estrogen Receptor (ER) to contribute to its cytotoxic activity on breast cancer cells. This study aimed to determine the potential of Cambodian leaf compounds and extracts as chemopreventive agents for luminal breast cancer with a molecular target of COX-2. Ethanol was used to extract the active compound of Cambodian leaves. The study on chemical activity against COX-2 employed molecular docking with Molecular Operating Environment (MOE) and the cytotoxic property of Cambodian leaf extract (CLE) on T47D was determined using the trypan blue exclusion method. The extraction yielded as 4.87% w/w CLE. Thin layer chromatography showed that Cambodian leaves contain sterol. Molecular docking confirmed that several sterol compounds have greater affinity to COX-2 than native ligands indicating that they are potent as COX-2 inhibitors. They are Stigmast-7-en-3-ol, Lupeol Acetate, and Lupeol carboxylic acid with docking scores of -14.3874, -13.8098, and -14.1045 kcal/mol respectively. The CLE exhibited cytotoxic activity on T47D cells with an IC50 value of 18 μg/mL. Therefore, CLE has a potential effect as a chemopreventive agent for breast cancer and potentially as a COX-2 inhibitor.Keywords: Cambodian leaf extract, breast cancer, COX-2 inhibitor, chemopreventive