Combinatorial Chemistry & High Throughput Screening - Volume 25, Issue 5, 2022

SARS-CoV-2, the novel coronavirus and the causative organism of the Covid-19 pandemic wreaked havoc worldwide producing asymptomatic to symptomatic cases leading to significant morbidity and mortality even after infection. Most of the countries reported a mortality rate of 2-3 % majorly due to cardiorespiratory failures. Recent studies highlighted the neurological involvement playing a key role in cardiorespiratory failures and other symptoms such as headache, anosmia, and ageusia observed in Covid-19 patients. Studies suggest SARS-CoV-2 entry via Olfactory Epithelium (OE), and the expression of type 2 transmembrane serine protease (TMPRSS2) in addition to Angiotensin-Converting Enzyme 2 (ACE2) can facilitate SARS-CoV-2 neurotropism. The virus can either travel via peripheral blood vessel causing endothelial dysfunction, triggering coagulation cascade and multiple organ dysfunction or reach the systemic circulation and take a different route to the Blood-Brain Barrier (BBB), disrupting the BBB causing neuroinflammation or neuronal excitotoxicity resulting in the development of encephalitis, encephalopathy, seizures, and strokes. SARS-CoV-2 invasion on the brain stem is believed to be responsible for the cardiorespiratory failures observed in Covid-19 patients. Apart from viral invasion via hematogenous route, SARS-CoV-2 neural invasion via PNS nerve terminal, results in viral replication and retrograde transportation to soma leading to invasion of the CNS including the brain producing neurological manifestations of the disease either in the initial stages or during the course of the disease and even for a long period post-infection in many cases. The ACE2 receptors are expressed in the brain and glial cells and SARS-CoV-2 acts via neuronal as well as nonneuronal pathways. But the exact cell types involved and how they can trigger inflammatory pathways need further in-depth study for the development of targeted therapy.

As most of the countries have already passed the peaks of the second or third wave of infections, COVID-19 is (finally) on the wane and will soon touch the baseline persistence with a reduced death rate. The Case Fatality Ratio (CFR) or lethality is presently only ~2.0%, which is relatively low in comparison to the high of 14% in April 2020. More importantly, the lethality in healthy people is 20 times lower. In addition, successful vaccination programs are currently underway in several countries. Thus, although new pathogenic variants as well as a few isolated cases of resurgence, will continue to emerge, the worst seems to be over and we will surely win this battle soon.

Background: Thiadiazoles can be considered as the privileged scaffold having diverse pharmacological potentials such as antihypertensive, anti-HIV, antimicrobials, antileishmanial agents, etc. In particular, 1,2,5-thiadiazoles and their fused analogues are subjects of fast-growing interest due to their higher significance in the fields of biomedicine and material sciences. Objective: This study aims to collect detailed medicinal information about aspects of 1,2,5- thiadiazole. Methods:A systemic search has been carried out using PubMed, Google Scholar, CNKI, etc., for relevant studies having the keyword, ‘1,2,5-thiadiazole’. Results and Conclusion: In this mini-review, we have covered known procedures of the synthesis and explored in details all known advancements of this scaffold concerning to its biological activities.

Background: Andrographis paniculata, commonly known as “Kalmegh”, is an annual herbaceous plant from the family Acanthaceae. The whole plant of A. paniculata has been explored for multiple pharmacological activities and is scientifically recognized by in-vivo and in-vitro studies. Various biotechnologically engineered techniques have been explored to enhance the bioavailability of this plant. Objective: In this review, we aim to present comprehensive recent advances in ethnopharmacology, phytochemistry, specific pharmacology, safety and toxicology, and bioavailability of A. paniculata and its pure compounds. Possible directions for future research are also outlined in brief, which will encourage advance investigations on this plant. Methods: Information on the recent updates of the present review is collected from different electronic scientific databases such as Science Direct, PubMed, Scopus, and Google Scholar. All the composed information is classified into different sections according to the objective of the paper. Results: More than a hundred research and review papers have been studied and incorporated in the present manuscript. After a vast literature search on A. paniculata, we present a noteworthy report of various phytoconstituents present in the plant, which are accountable for the potential therapeutic properties of the plant. Forty-five of the studied articles gave general information about the introduction, ethnobotany, and traditional uses of the plant. Twenty-two papers enclosed information about the phytoconstituents present in different parts of A. paniculata and seventy-two papers briefly outlined the pharmacological activities like antioxidant, anti-dengue, anti-ulcerogenic, antifungal, some miscellaneous activities like activity against SARS-CoV-2, antidiarrhoeal. Nineteen studies highlighted the research work conducted by various researchers to increased the bioavailability of A. Paniculata and two studies reported the safety and toxicology of the plant. Conclusion: This review incorporated the scientifically validated research work encompassing the ethnobotanical description of the subjected plant, phytochemical profile, various pharmacological activities, and recent approaches to enhance the bioavailability of active metabolites.

Natural bioactive compounds with anti-carcinogenic activity are gaining tremendous interest in the field of oncology. Cinnamon, an aromatic condiment commonly used in tropical regions, appeared incredibly promising as an adjuvant for cancer therapy. Indeed, its whole or active parts (e.g., bark, leaf) exhibited significant anti-carcinogenic activity, which is mainly due to two cinnamaldehyde derivatives, namely 2-hydroxycinnaldehyde (HCA) and 2- benzoyloxycinnamaldehyde (BCA). In addition to their anti-cancer activity, HCA and BCA exert immunomodulatory, anti-platelets, and anti-inflammatory activities. The highly reactive α,ßunsaturated carbonyl pharmacophore, called Michael acceptor, contributes to their therapeutic effects. The molecular mechanisms underlying their anti-tumoral and anti-metastatic effects are miscellaneous, strongly suggesting that these compounds are multi-targeting compounds. Nevertheless, unravelling the exact molecular mechanisms of HCA and BCA remains a challenging matter which is necessary for optimal controlled-drug targeting delivery, safety, and efficiency. Eventually, their poor pharmacological properties (e.g., systemic bioavailability and solubility) represent a limitation and depend both on their administration route (e.g., per os, intravenously) and the nature of the formulation (e.g., free, smart nano-). This concise review focused on the potential of HCA and BCA as adjuvants in cancer. We describe their medicinal effects as well as provide an update about their molecular mechanisms reported either in-vitro, ex-vivo, or in animal models.

Background: Papillary thyroid carcinoma (PTC) is a subtype of thyroid cancer with increasing incidence over time. Objective: This study aimed to build a risk score (RS) system for PTC patients. Methods: PTC microRNA (miRNA) and messenger RNA (mRNA) expression data were extracted from The Cancer Genome Atlas (TCGA) database. The 491 PTC samples were randomly divided into training and validation sets. Using the limma software package, differentially expressed mRNAs (DEGs) and miRNAs (DEMs) between the tumor and control groups were screened. In order to construct an RS system, a survival package was used to select independent miRNAs related to prognosis. Enrichment analysis was performed, and a miRNA-mRNA co-expression network was constructed. High-throughput sequencing was also used to verify the prognostic miRNAs in exosomes. Results: We found 1363 DEGs and 171 DEMs between the tumor and control groups. After identifying 26 DEMs significantly related to prognosis, 6 independent prognosis-associated miRNAs were selected to build an RS system. The areas under the curves of the overall survival rates of the training, validation, and entire sets were 0.847, 0.772, and 0.819, respectively. By conducting pathway analysis using the miRNA-mRNA co-expression network, one overlapping factor and five overlapping pathways were obtained. In addition, high-throughput sequencing revealed that the hsa-miR-129-2, hsa-miR-548j, hsa-miR-6734, and hsa-miR-889 expression levels in TCGA tumor tissues and exosomes were consistent, and those of hsa-miR-129-2 and hsa-miR- 889 between patients and controls were significantly different in exosomes. Conclusion: The six-miRNA RS system in exosomes may comprise independent signatures for predicting PTC patient prognosis.

Background: Deubiquitinating enzymes (DUBs) protein family have been implicated in some deregulated pathways involved in carcinogeneses, such as cell cycle, gene expression, and DNA damage response (DDR). Zinc finger with UFM1-specific peptidase domain protein (ZUFSP) is one of the recently discovered members of the DUBs. Objectives: To identify and cross-validate the ZUFSP binding site using the bioinformatic tools, including SiteMap&Metapocket, respectively. To understand the molecular basis of complementary ZUFSP-Ub interaction and associated structural events using MD Simulation. Methods: In this study, four binding pockets were predicted, characterized, and cross-validated based on physiochemical features such as site score, druggability score, site volume, and site size. Also, a molecular dynamics simulation technique was employed to determine the impact of ubiquitin-binding on ZUFSP. Results: Site 1 with a site score 1.065, Size 102, D scores 1.00, and size volume 261 was predicted to be the most druggable site. Structural studies revealed that upon ubiquitin-binding, the motional movement of ZUFSP was reduced when compared to the unbound ZUFSP. Also, the ZUFSP helical arm (ZHA) domain orient in such a way that it moves closer to the Ub; this orientation enables the formation of a UBD which is very peculiar to ZUFSP. Conclusion: The impact of ubiquitin on ZUFSP movement and the characterization of its predicted druggable site can be targeted in the development of therapeutics.

Background: This paper aims to reveal an urgent industrial scheme for a fast and facile total synthesis of umifenovir (arbidol) (by one-pot stages) as an antiviral agent for treating the 2019-nCoV virus via inhibiting its viral replication in the human cells. As COVID-19 takes thousands of lives all around the world, it seems that the medicinal resources would not be enough to supply billions of peoples currently living on the planet. Thus, this pandemic and its subsequent impacts on the natural order of our life would be one of the most important threats against the entire human race. Methods: In this project, we have made attempts to find an operative approach for synthesizing this compound as an active pharmaceutical ingredient (API), which showed it could be effective in inhibiting the newly emerged coronavirus.. Results: The designed scheme uses relatively cheap precursors and contains one pot stage instead of seven time-consuming and more costly linear steps. Moreover, safe and cheap solvents have been used like water and ethanol, instead of toxic ones like methanol and pyridine which could cause rejection of the API in the organic volatile impurities (OVI) test of pharmacopeia analysis, as well as increase the concern of inflammability, explosivity, and carcinogenic properties of those common solvents. Conclusion: The most important pharmaceutical analytical methods containing OVI test (mainly ethanol (about 171 ppm) much lower than the limits, by gas chromatography-Flame Ionization Detector (GC-FID) instrument), assay content (about 99.6% by potentiometric titration), and related purity analysis (by high-performance liquid chromatography-Ultraviolet Detector (HPLCUV)) (about 99.8%) were performed and described to give a more clear industrial scheme.

Background: Cognitive impairment is a common neurocognitive disorder that affects the health of millions of people worldwide, related to folate deficiency. Objective: The present study aimed to investigate the lncRNA-mRNA functional networks associated with cognitive impairment in folate-deficient mice and elucidate their possible molecular mechanisms. Methods: We downloaded the gene expression profile (GSE148126) of lncRNAs and mRNAs from NCBI Gene Expression Omnibus (GEO) database. Four groups of mouse hippocampi were analyzed, including 4 months (4mo) and 18 months (18mo) of folic acid (FA) deficiency/supplementation. The differentially expressed lncRNAs (DElncRNAs) and mRNAs (DEmRNAs) were identified using gplots and heatmap packages. The functions of the DEmRNAs were evaluated using Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. The hub genes were identified by CytoHubba plugins of Cytoscape, and protein-protein interaction (PPI) network of deregulated mRNAs was performed using the STRING database. Finally, lncRNA-mRNA co-expression and competitive endogenous RNA (ceRNA) network analyses were constructed. Results: In total, we screened 67 lncRNAs with 211 mRNAs, and 89 lncRNAs with 229 mRNAs were differentially expressed in 4mo_FA and 18mo_FA deficient mice, respectively. GO analyses indicated that DEmRNAs were highly related to terms involved in binding and biological regulation. KEGG pathway analyses demonstrated that these genes were significantly enriched for renin secretion, pancreatic secretion, and AMPK signaling pathways in the 18mo_FA deficiency group. Subsequently, the top 5 hub genes were screened from the PPI network, which may be key genes with the progression of folate deficiency. Upon the lncRNA-mRNA co-expression network analysis, we identified the top 10 lncRNAs having the maximum number of connections with related mRNAs. Finally, a ceRNA network was constructed for DE lncRNAs and DEmRNAs, and several pivotal miRNAs were predicted. Conclusions: This study identified the lncRNA-mRNA expression profiles and functional networks associated with cognitive impairment in folate-deficient mice by bioinformatics analysis, which provided support for the possible mechanisms and therapy for this disease.

Aim and Objective: Europium (Eu(III)) is a rare-earth metal, the softest, least dense, and most volatile member of lanthanides. It is greatly applied in the control rods of nuclear reactors. Although various extraction methods of Eu(III) have been reported, we present a novel mixture of easily available extractants in optimized experimental conditions to extract it efficiently, quickly, and cost-effectively. Materials and Methods: Physical-chemical conditions (e.g., pH, equilibration time, temperature, europium concentration, extractants concentration, presence of specific metal ions) were optimized. The extractants, picrolonic acid (HPA) and di-n-butylsulfoxide (DBSO), were thoroughly mixed at equal concentration in chloroform. Standard Eu(III) solution was used for determining the method’s accuracy. Reagent blank was prepared under identical conditions but without metal ions. Using the metallochromic dye arsenazo III as the blank, the absorbance of Eu(III) was measured spectrophotometrically at 651 nm. The distribution ratio (i.e., Eu(III) concentration in the aqueous phase before and after extraction) defined the extraction yield. Results: HPA/DBSO mixture (0.01 M) had a synergistic effect on Eu(III) extraction (1.19×10^-5 mole/dm³), achieving a maximum yield (≥ 99%) at pH 2, during 5 minutes equilibration at room temperature. Eu(III) extraction was reduced depending on the nature but not on the metal ions concentration. Extractants could be recycled four times without consequent degradation. Deionized water (dH2O) was the best strippant besides its availability and low-cost. The composition of the extracted adduct was defined as Eu(PA)3.2DBSO. Conclusion: This alternative method was found to be stable, simple, rapid, cost-effective, reliable, accurate and sensitive. It could be used for Eu(III) extraction and refining on a pilot plant scale.

Background: Due to side-effects and low efficacy of common drugs on new resistant strains of Mycobacterium tuberculosis (Mtb), investigation on novel drugs and natural compounds from rich sources of endemic plants is required. Thus, in the present study, the anti-mycobacterial effects of 22 Iranian endemic or rare plant extracts on multi-drug resistance (MDR) and extensively-drug resistance (XDR) Mtb isolates were evaluated. Methods: Twenty-two Iranian endemic and rare plant species from 9 families were collected and extracted by methanol. Their inhibitory-effects were then evaluated against Mtb H37Rv strain, seven clinical MDR-TB, and two XDR-TB isolates using the resazurin microtiter assay (REMA) method. The best of them were then fractionalized by five different polar solvents (Petroleum- Ether, Dichloromethane, Ethyl-Acetate, n-butanol, and water). To find anti-mycobacterial fractions, the inhibitory effect of isolated fractions was tested on Mtb H37Rv. Results: Out of the 22 plants, 14 plant extracts demonstrated anti-mycobacterial activity with minimum inhibitory concentration (MIC) ranging from 4 to 30μg/mL against Mtb H37Rv. Eight plant extracts also exhibited anti-mycobacterial activity against MDR and XDR clinical strains of Mtb by MICs, i.e., 15-60μg/ml. Crinitaria grimmii and Linum album were the best antimycobacterial plants. Among fractions of Crinitaria grimmii, dichloromethane and n-butanol, and for Linum album, dichloromethane and Ethyl-Acetate fractions displayed more anti-mycobacterial effect as compared to crude extract on Mtb. Conclusion: The present study confirms the potential role of some plants to treat respiratory diseases as our results have demonstrated that these plants exhibit anti-mycobacterial activity in the acceptable range against Mtb. Thus, these plants could be good sources and alternatives of plant metabolites for anti-TB-drug development.

Background: Diabetic retinopathy (DR) is a severe complication of diabetes; however, the pathogenesis of DR has not been completely clarified, which is mostly dependent on the molecular pathology. This study aimed to investigate key serum-derived miRNAs associated with DR. Methods: miRNA expression profile arrays of human umbilical vein endothelial cells (HUVECs) treated with glucose were downloaded from the Gene Expression Omnibus (GEO) database (GSE74296). Weighted gene co-expression network analysis (WGCNA) was performed to obtain hub miRNAs, which were verified in HUVECs treated with 40 mM and 5 mM glucose, respectively. Meanwhile, serum samples of patients with DR and healthy controls were collected, and EVs were extracted from the patients’ serum by ultracentrifugation. Hub miRNAs associated with endothelial dysfunction were verified in healthy individuals before and after treatment of patients with DR, by qRT-PCR. Results: These miRNAs were categorized into six modules, among which miR-26b-5p had a strong association with other modules. This miRNA was also one of the hyperglycemia-induced miRNAs related to endothelial dysfunction. miR-26b-5p was up-regulated in HUVECs treated with 40 mM glucose and in the serum of patients with DR before and after treatment. Furthermore, miR- 26b-5p was slightly up-regulated in serum-derived EVs but not in serum without EVs in DM patients. Conclusion: Our results suggest that EVs derived from miR-26b-5p are up-regulated in the serum of patients with DR.

Background: This research proposes the application of a novel photocatalyst including Co3O4@SiO2@TiO2-Ag nanocomposite with highly photocatalytic stability and core-shell structure for the removal of toxic methyl violet from aqueous solution. Objective: The removal of toxic dyes and organic contaminants from water is an outstanding research area among the scientists. Methyl violet is a toxic cationic pollutant that has a disruptive influence on humans. In this research, with an aim to remove methyl violet from the wastewater, we developed a new photocatalyst including Co3O4@SiO2@TiO2-Ag nanocomposite as an ecofriendly and low-cost nanostructure with high photocatalytic activity in order to reduce the risks of this pollutant from aqueous media. Methods: The Co3O4@SiO2@TiO2-Ag nanostructure was prepared via hydrothermal and sol-gel methods and the structure elucidation of the prepared photocatalyst was analyzed by different spectroscopy techniques, including XRD, FT-IR, FE-SEM, TEM, VSM and EDX. Results: Photodegradation of methyl violet in the presence of different structures showed that Co3O4@SiO2@TiO2-Ag possesses superior photocatalytic activity (about 98% decomposed after 40 min) compared to the previous shells and pure Co3O4 NPs. Loadings of SiO2@TiO2-Ag nanocomposite over the Co3O4 surface led to the reduction in the bandgap energy of visible light and improvement in the photocatalytic activity of Methyl Violet dye f o r the aqueous phase decomposition. Conclusion: The remarkable benefits of this nanocomposite are high photocatalytic efficiency in the degradation of methyl violet (almost 100 % within 1 h), easy magnetic separation, low cost, and high chemical stability. The collected results demonstrated that the rate of degradation increased by increasing the irradiation time, while the rate of degradation decreased with increasing dye concentration.

Background: Frankincense is a resin secreted by the Boswellia tree. It is used in perfumery, aromatherapy, skincare, and traditional Chinese medicine. However, all Boswellia species are under threat owing to habitat loss and overexploitation. As a result, the market is getting flooded with counterfeit frankincense products. Objective: This study aims to establish a high-throughput method to screen and identify the authenticity of commercial frankincense products. We report, for the first time, a matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS)-based method for rapid and high-throughput screening of frankincense samples. Methods: MALDI-TOF MS, HPLC, thin-layer chromatography (TLC), and in vitro antiinflammatory activity assay were used to examine the frankincense samples. Results: Well-resolved peaks of frankincense triterpenoids in the spectra were observed in the crude extract of commercial samples, including α-boswellic acids (αBAs), β-boswellic acids (βBAs), 11-keto-β-boswellic acids (KBAs), acetyl-11-keto-β-boswellic acids (AKBAs), and their esters. These compounds can be used as indicators for determining the authenticity of frankincense. Conclusion: Unlike LC–MS, which is a time-consuming and expensive method, and TLC, which requires a reference sample, our inexpensive, rapid high-throughput identification method based on MALDI-TOF MS is ideal for large-scale screening of frankincense samples sold in the market.

Background: Hepatocellular carcinoma (HCC) is a malignant tumour with a poor prognosis. The effect of DNA repair on prognosis cannot be ignored, and long non-coding RNA (lncRNA) can regulate the DNA repair process. Objective: To obtain DNA repair-associated lncRNA (DR-lncRNA) prognostic signature for improving the ability to predict HCC prognosis. Methods: Our study used the Cancer Genome Atlas database. Gene set variation analysis was performed to differentiate high and low levels of DNA repair to identify DR-lncRNAs. By performing univariate Cox regression, LASSO regression, and multivariate Cox regression analyses, we finally obtained a DR-lncRNA prognostic signature and constructed a nomogram prognostic model. Time-dependent receiver operating characteristic (ROC) curves, calibration plots, decision curve analysis (DCA), and clinical impact curves were used to assess predictive ability and clinical utility. Differentially expressed genes (DEGs) functional enrichment analysis was performed to further explore the underlying mechanisms that influence HCC prognosis. Results: We obtained the following DR-lncRNA prognostic signature:AP002478.1, AC116351.1, LINC02580, and LINC00861. The ROC curves and calibration plots showed good discrimination and calibration properties. Combining the DR-lncRNA prognostic signature and tumour stages, we established a nomogram prognostic model. DCA and clinical impact curves showed the clinical utility of this model. DEGs of high-risk and low-risk groups predicted by the prognostic DRlncRNA were significantly associated with cell cycle, various metabolic pathways and biological processes, such as the oxidation-reduction process and cell division. Conclusion: We identified a DR-lncRNA prognostic signature and constructed a nomogram prognostic model, which could be a beneficial prognostic strategy for HCC.

Aim: The aim of this study was to investigate the effect and molecular mechanism of Scutellaria baicalensis Georgi stems and leaves flavonoids (SSF) in promoting neurogenesis and improving memory impairment induced by the PI3K-AKT-CREB signaling pathway. Methods: Alzheimer's disease (AD) was induced in the male Wistar rats by intracerebroventricular injection of amyloid beta peptide 25-35 (Aβ25-35) in combination with aluminum trichloride (AlCl3) and recombinant human transforming growth factor-β1(RHTGF-β1) (composited Aβ). The Morris water maze was used to screen the successful establishment of the memory impairment model of rats. The screened successful model rats were randomly divided into a model group and three groups of three different doses of the drug (SSF). Rats in the drug group were treated with 35, 70, and 140 mg/kg of SSF for 43 days. The Eight-arm maze was used to measure the spatial learning and memory abilities of the rat, including working memory errors (WME) and reference memory errors (RME). Immunohistochemistry was used to detect the expression of BrdU, an indicator of neuronal proliferation, in the hippocampal gyrus of rats. The mRNA and protein expressions of TRKB, PI3K, AKT, P-AKT, and IGF2 in the PI3K-AKT-CREB signaling pathway in the hippocampus and cerebral cortex of the rats were determined by quantitative real-time PCR (qPCR) and Western blotting methods. Results: Compared to the sham group, the spatial memory ability of rats with composited Aβ was decreased, the number of WME and RME (P < 0.01) was increased, the expression of BrdU protein (P < 0.01) in the hippocampal gyrus was reduced, the mRNA and protein expression levels of TRKB, AKT, and IGF2 (P < 0.01, P < 0.05) in the hippocampus and cerebral cortex were lowered, and the mRNA expression level of PI3K (P < 0.01) in the cerebral cortex and the protein expression level of PI3K (P < 0.01) in the hippocampus were augmented. However, compared to the model group, the three-doses of SSF improved memory disorder induced by composited Aβ, reduced the number of WME and RME, increased the expression of BrdU protein in the hippocampal gyrus, and differently regulated the mRNA and protein expressions in composited Aβ rats. Conclusion: SSF improved memory impairment and neurogenesis disorder induced by composited Aβ in rats by activating the PI3K-AKT-CREB signaling pathway and up-regulating the mRNA and protein expressions of TRKB, PI3K, AKT, CREB, and IGF2.

Aim and Objective: To investigate the effect of Polyphyllin I (PPI) on HBV-related liver cancer through network pharmacology and in vitro experiments, and to explore its mechanism of action. Materials and Methods: Use bioinformatics software to predict the active ingredient target of PPI and the disease target of liver cancer, and perform active ingredient-disease target analysis. The results of network pharmacology through molecular docking and in vitro experiments can be further verified. The HepG2 receptor cells (HepG2. 2. 15) were transfected with HBV plasmid for observation, with the human liver cancer HepG2 being used as the control. Results: Bioinformatics analysis found that PPI had a total of 161 protein targets, and the predicted target and liver cancer targets were combined to obtain 13 intersection targets. The results of molecular docking demonstrated that PPI had a good affinity with STAT3, PTP1B, IL2, and BCL2L1. The results of the in vitro experiments indicated that the PPI inhibited cell proliferation and metastasis in a concentration-dependent manner (P<0.01). Compared with the vehicle group, the PPI group of 1.5, 3, and 6 μmol/L can promote the apoptosis of liver cancer to different degrees (P<0.01). Conclusion: The present study revealed the mechanism of PPI against liver cancer through network pharmacology and in vitro experiments. Its mechanism of action is related to the inhibition of PPI on the proliferation of HBV-related liver cancer through promoting the apoptosis of liver cancer cells. Additionally, in vitro experiments have also verified that PPI can promote the apoptosis of HepG2 and HepG2.2.15 cells.