To explore the pharmacological mechanism of action using digital twin

With the advent of medical technology and science, the number of animals used in research has increased. For decades, the use of animals in research and product testing has been a point of conflict. Experts and pharmaceutical manufacturers are harming animals worldwide during laboratory research. Animals have also played a significant role in the advancement of science; animal testing has enabled the discovery of various novel drugs. The misery, suffering, and deaths of animals are not worth the potential human benefits. As a result, animals must not be exploited in research to assess the drug mechanism of action (MOA). Apart from the ethical concern, animal testing has a few more downsides, including the requirement for skilled labor, lengthy processes, and cost. Because it is critical to investigate adverse effects and toxicities in the development of potentially viable drugs. Assessment of each target will consume the range of resources as well as disturb living nature. As the digital twin works in an autonomous virtual world without influencing the physical structure and biological system. Our proposed framework suggests that the digital twin is a great reliable model of the physical system that will be beneficial in assessing the possible MOA prior to time without harming animals. The study describes the creation of a digital twin to combine the information and knowledge obtained by studying the different drug targets and diseases. Mechanism of Action using Digital twin (MOA-DT) will enable the experts to use an innovative approach without physical testing to save animals, time, and resources. DT reflects and simulates the actual drug and its relationships with its target, however presenting a more accurate depiction of the drug, which leads to maximize efficacy and decrease the toxicity of a drug. In conclusion, it has been shown that drug discovery and development can be safe, effective, and economical in no time through the combination of the digital and physical models of a pharmaceutical as compared to experimental animals.

Comparison between Heat-Clearing Medicine and Antirheumatic Medicine in Treatment of Gastric Cancer Based on Network Pharmacology, Molecular Docking, and Tumor Immune Infiltration Analysis

Background. Clinical research found that TCM is therapeutic in treating gastric cancer. Clearing heat is the most common method, while some antirheumatic medicines are widely used in treatment as well. To explore the pharmacological mechanism, we researched the comparison between heat-clearing medicine and antirheumatic medicine in treating gastric cancer. Methods. First, related ingredients and targets were searched, respectively, and are shown in an active ingredient-target network. Combining the relevant targets of gastric cancer, we constructed a PPI network and MCODE network. Then, GO and KEGG enrichment analyses were conducted. Molecular docking experiments were performed to verify the affinity of targets and ligands. Finally, we analyzed the tumor immune infiltration on gene expression, somatic CNA, and clinical outcome. Results. A total of 31 ingredients and 90 targets of heat-clearing medicine, 31 ingredients and 186 targets of antirheumatic medicine, and 12,155 targets of gastric cancer were collected. Antirheumatic medicine ranked the top in all the enrichment analyses. In the KEGG pathway, both types of medicines were related to pathways in cancer. In the KEGG map, AR, MMP2, ERBB2, and TP53 were the most crucial targets. Key targets and ligands were docked with low binding energy. Analysis of tumor immune infiltration showed that the expressions of AR and ERBB2 were correlated with the abundance of immune infiltration and made a difference in clinical outcomes. Conclusions. Quercetin is an important ingredient in both heat-clearing medicine and antirheumatic medicine. AR signaling pathway exists in both types of medicines. The mechanism of the antitumor effect in antirheumatic medicine was similar to trastuzumab, a targeted drug aimed at ERBB2. Both types of medicines were significant in tumor immune infiltration. The immunology of gastric tumor deserves further research.

Network pharmacology approach to reveals therapeutic mech-anism of traditional plants formulation used by Malaysia in-digenous ethnics in coronaviruses infection

Network pharmacology analysis can act as a strategy to identify the pharmacological effect of plant-based bioactive compounds against coronavirus diseases. This study aimed to investigate the potential pharmacological mechanism of a local ethnomedicine (Costus speciosus, Hibiscus rosa-sinensis and Phyllanthus niruri) of Northern Borneo against coronaviruses known as CHP. Compounds in CHP were extracted from databases and screened for their oral bioavailability and drug-likeness before a compound-target network was built. Furthermore, the protein-protein interaction network and pathway enrichment were constructed and analyzed. A compound-target network consisting of 48 putative bioactive compounds targeting 587 candidate genes was identified. A total of 186 coronavirus-related genes were extracted and TP53, STAT3, HSP90AA1, STAT1, and EP300 were predicted to be the key targets. Notably, mapping of these target genes into the target-pathway network illustrated that functional enrichment was on viral infection and regulation of inflammation pathways. Urinatetralin is predicted, for the first time, as a bioactive compound that solely targets STAT3. The results from this study indicate that compounds present in CHP employ STAT3 and its connected pathways as the mechanism of action against coronaviruses. In conclusion, urinatetralin should be further investigated for its potential application against coronavirus infections.

A Study of the Protective Effect of Bushen Huoxue Prescription on Cerebral Microvascular Endothelia Based on Proteomics and Bioinformatics

Diabetic cognitive dysfunction is a serious complication of type 2 diabetes mellitus (T2DM), which can cause neurological and microvascular damage in the brain. At present, there is no effective treatment for this complication. Bushen Huoxue prescription (BSHX) is a newly formulated compound Chinese medicine containing 7 components. Previous research indicated that BSHX was neuroprotective against advanced glycosylation end product (AGE)-induced PC12 cell insult; however, the effect of BSHX on AGE-induced cerebral microvascular endothelia injury has not been studied. In the current research, we investigated the protective effects of BSHX on AGE-induced injury in bEnd.3 cells. Our findings revealed that BSHX could effectively protect bEnd.3 cells from apoptosis. Moreover, we analyzed the network regulation effect of BSHX on AGE-induced bEnd.3 cells injury at the proteomic level. The LC-MS/MS-based shotgun proteomics analysis showed BSHX negatively regulated multiple AGE-elicited proteins. Bioinformatics analysis revealed these differential proteins were involved in multiple processes, such as Foxo signaling pathway. Further molecular biology analysis confirmed that BSHX could downregulate the expression of FoxO1/3 protein and inhibit its nuclear transfer and inhibit the expression of downstream apoptotic protein Bim and the activation of caspase, so as to play a protective role in AGE-induced bEnd.3 injury. Taken together, these findings demonstrated the role of BSHX in the management of diabetic cerebral microangiopathy and provide some insights into the proteomics-guided pharmacological mechanism study of traditional Chinese Medicine.

Hepatoprotective activity of the ethanolic extract of Morus indica roots from Indian Bodo tribes

The roots of Morus species are well described in the Pharmacopoeia of the People's Republic of China (ChP) for its traditional use in treating liver fibrosis due to its hepatoprotective property. However, little is known about the hepatoprotective effect of the roots of Morus indica L. (RoMi), and the pharmacological mechanism(s) are uncertain due to its intricacy. Therefore, this study evaluates the hepatoprotective activity of the ethanolic extract of RoMi (eRoMi) against the CCl4-induced in-vivo animal model at different dosages (100 and 200 mg/kg BW) in comparison with silymarin as a positive control. The hepatoprotective activity of eRoMi was evaluated by measuring the levels of serum biomarkers, hepatic antioxidant enzymes and was verified by histological studies. Interestingly, 1,2-bis(trimethylsilyl) benzene, 1,4-phenylenebis (trimethylsilane), 2,4,6-cycloheptatriene-1-one, 3,5-bis-trimethylsilyl and α-amyrin were the active components found in eRoMi as detected by GC–MS. Oral administration of eRoMi (200 mg/kg BW) to rats significantly protected serum biochemical parameters (increased ALT, AST, LDH, bilirubin and GGT as well as depletion of antioxidant enzymes and hepatic GSH) and elevation in hepatic lipid peroxidation as compared to CCl4-treated rats. The hematological indices such as erythrocytes, hemoglobin, monocytes and lymphocytes were also normal in eRoMi-treated rats. The histopathological evaluation indicated a significant restoration of liver structure as compared to silymarin. This study is the first scientific validation for the traditional use of eRoMi to understand its hepatoprotective activity.

Identification the Hub Genes of Hirudin Intervention of Diabetic Nephropathy Based On Bioinformatic Analysis and Molecular Docking

Background: Diabetic nephropathy (DN) is one of the common complications of diabetes, it can cause a disproportionate burden. Leeches are widely used to treat DN in China, and hirudin is the main component of leeches. However, its pharmacological mechanisms and molecular targets are unclear. This work aimed to explore new biomarkers of DN and reveal the mechanism of hirudin in DN. Methods: Expression microarray datas between kidney tissues of DN and control individuals were obtained from the GEO database, and differentially expressed genes were identified as DN-related targets using the robust rank aggregation analysis. The SEA, GeneCards and SwissTargetPrediction databases were used to predict targets of hirudin. A protein-protein interaction network of DN-hirudin was conducted by Cytoscape software. GO and KEGG pathway enrichment analyses were carried out to explore the involved pharmacological mechanism of hirudin in treatment of DN. And molecular docking was adopted to predict the hub targets of hirudin. Results: A total of 30 significant DEGs (16 up- and 14 down-regulated) were identified. ATF3, SLC22A8 and TGF-Β1 are likely as new biomarkers of DN. The 42 candidate targets were identifyed in PPI network. GO analysis revealed that these targets were enriched with ubiquitin protein ligase, transcription factor binding and nuclear transport. The KEGG pathways were enriched, including AGE-RAGE signaling pathway in diabetic complications, PI3K-Akt signaling pathway, Focal adhesion. The docking results showed that hirudin could easily dock with ITGA4, EGFR and ESR1. Conclusion: Our study demonstrates that hirudin is involved in DN therapy through a multi-targeted, multi-pathway approach. It provides a basis for further research on its mechanism of action.

Network Pharmacology-Based Investigation and Experimental Exploration of the Antiapoptotic Mechanism of Colchicine on Myocardial Ischemia Reperfusion Injury

Background: The beneficial effects of colchicine on cardiovascular disease have been widely reported in recent studies. Previous research demonstrated that colchicine has a certain protective effect on ischemic myocardium and has the potential to treat myocardial ischemia reperfusion injury (MIRI). However, the potential targets and pharmacological mechanism of colchicine to treat MIRI has not been reported.Methods: In this study, we used network pharmacology and experimental verification to investigate the pharmacological mechanisms of colchicine for the treatment of MIRI. Potential targets of colchicine and MIRI related genes were screened from public databases. The mechanism of colchicine in the treatment of MIRI was determined by protein-protein interaction (PPI), gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Additionally, we evaluated the effect of colchicine on H9C2 cell activity using CCK-8 assays, observed the effect of colchicine on H9C2 cell apoptosis via flow cytometry, and further verified the expression of key targets after colchicine treated by Western blot.Results: A total of 626 target genes for colchicine and 1549 MIRI disease targets were obtained. 138 overlapping genes were determined as potential targets of colchicine in treating MIRI. the PPI network analysis demonstrated that the targets linked to MIRI were ALB, TNF, ACTB, AKT1, IL6, TP53, IL1B, CASP3 and these targets showed nice affinity with colchicine in molecular docking experiments. The results of GO analysis and KEGG pathway enrichment demonstrated that the anti-MIRI effect of colchicine involves in apoptotic signaling pathway. Further tests suggested that colchicine can protect H9C2 cell from Hypoxia/Reoxygenation (H/R) injury through anti-apoptotic effects. Western blot results demonstrated that colchicine can inhibited MIRI induced apoptosis of H9C2 cell by enhancing the decreased levels of Caspase-3 in myocardial injure model induced by H/R and activating the PI3K/AKT/eNOS pathway.Conclusions: we performed network pharmacology and experimental evaluation to reveal the pharmacological mechanism of colchicine against MIRI. The results from this study could provide a theoretical basis for the development and clinical application of colchicine.