Biochemical characterization of bioinspired nanosuspensions from Swertia chirayita extract and their therapeutic effects through nanotechnology approach.

Swertia chirayita is used as a traditional medicinal plant due to its pharmacological activities, including antioxidant, antidiabetic, antimicrobial, and cytotoxic. This study was aimed to evaluate the therapeutic efficacy of newly synthesized nanosuspensions from Swertia chirayita through nanotechnology for enhanced bioactivities. Biochemical characterization was carried out through spectroscopic analyses of HPLC and FTIR. Results revealed that extract contained higher TPCs (569.6 ± 7.8 mg GAE/100 g)) and TFCs (368.5 ± 9.39 mg CE/100 g) than S. chirayita nanosuspension, TPCs (500.6 ± 7.8 500.6 ± 7.8 mg GAE/100 g) and TFCs (229.5± 3.85 mg CE/100 g). Antioxidant activity was evaluated through DPPH scavenging assay, and nanosuspension exhibited a lower DPPH free radical scavenging potential (06 ±3.61) than extract (28.9± 3.85). Anti-dabetic potential was assessed throughα-amylase inhibition and anti-glycation assays. Extract showed higher (41.4%) antiglycation potential than 35.85% nanosuspension and 19.5% α-amylase inhibitory potential than 5% nanosuspension. Biofilm inhibition activity against E. coli was higher in nanosuspension (69.12%) than extract (62.08%). The extract showed high cytotoxicity potential (51.86%) than nanosuspension (33.63%). These nanosuspensions possessed enhanced bioactivities for therapeutic applications could be explored further for the development of new drugs.

Metabolomics-based pharmaceutical evaluation of different parts of Swertia chirayita (Roxb.) Buch.-Ham. ex C.B. Clarke from the western Himalayas.

Swertia species are common ingredients in numerous herbal remedies. It is also used to treat a wide range of illnesses and possess diverse therapeutic activities. The aim of the study is to elucidate the comprehensive metabolomics profile of Swertia chirayita and the role of various extraction methods in the phytochemical compositions of the extracts of S. chirayita, and their antioxidant and enzyme inhibitory activities. Extraction of the stems, leaves, and flowering tops of S. chirayita was performed by maceration, infusion, and soxhlation using methanol and water as solvent. Extracts were subjected to phytochemical profiling by a liquid-chromatographic system. Antioxidant and enzyme inhibitory activity was carried out. The metabolomics profiling showed that a diverse range of specialized metabolites were present in the stems and leaves & flowering tops of the plant. All the extracts showed substantial antioxidant and enzyme inhibitory activities further confirmed by molecular docking studies. This study appraised the use of S. chirayita aerial parts as a potential antioxidant and its therapeutic application in various chronic illnesses including Alzheimer's disease, diabetes, and other skin-related disorders.

Multidirectional insights into the phytochemical, biological, and multivariate analysis of extracts from the aerial part of Swertia perennis Linnaeus.

Swertia perennis Linnaeus (SP) has been utilised to treat gastritis. We report the qualitative and quantitative phytochemical analysis, antioxidant and enzyme inhibitory activities of SP. The correlation between the biological activities and total bioactive contents of the extracts was also studied via multivariate analysis. Methanol extract contained many active compounds and exhibited good antioxidant activity. Therefore, this was selected for further phytochemical profiling and stability studies. Fourteen compounds were identified by ultra-performance liquid chromatography-electrospray ionisation-orbitrap-mass spectrometry for the first time from this plant. Iridoids, xanthones, and flavonoids were the main components. Methanol extract exhibited good stability and antioxidant capacity in stability studies, with low toxicity, and showed a protective effect on the oxidation of olive and sunflower oils. SP has the potential to be developed and used as an antioxidant, or as urease and XO inhibitors, and its methanol extract could be used as a natural oil stabiliser.

Synergistic Influence of Yeast Extract and Calcium Oxide Nanoparticles on the Synthesis of Bioactive Antioxidants and Metabolites in Swertia chirata In Vitro Callus Cultures.

The challenges in the production of metabolites of medicinal potential from wild plants include low yields, slow growth rates, seasonal variations, genetic variability and regulatory as well as ethical constraints. Overcoming these challenges is of paramount significance and interdisciplinary approaches and innovative strategies are prevalently applied to optimize phytoconstituents' production, enhance yield, biomass, ensure sustainable consistency and scalability. In this study, we investigated the effects of elicitation with yeast extract and calcium oxide nanoparticles (CaONPs) on in vitro cultures of Swertia chirata (Roxb. ex Fleming) Karsten. Specifically, we examined the effects of different concentrations of CaONPs in combination with different concentrations of yeast extract on various parameters related to callus growth, antioxidant activity, biomass and phytochemical contents. Our results showed that elicitation with yeast extract and CaONPs had significant effects on the growth and characteristics of callus cultures of S. chirata. The treatments involving yeast extract and CaONPs were found to be the most effective in increasing the contents of total flavonoid contents (TFC), total phenolic contents (TPC), amarogentin and mangiferin. These treatments also led to an improvement in the contents of total anthocyanin and alpha tocopherols. Additionally, the DPPH scavenging activity was significantly increased in the treated samples. Furthermore, the treatments involving elicitation with yeast extract and CaONPs also led to significant improvements in callus growth and characteristics. These treatments promoted callus response from an average to an excellent level and improved the color and nature of the callus from yellow to yellow-brown and greenish and from fragile to compact, respectively. The best response was observed in treatments involving 0.20 g/L yeast extract and 90 ug/L CaONPs. Overall, our findings suggest that elicitation with yeast extract and CaONPs can be a useful strategy for promoting the growth, biomass, phytochemical contents and antioxidant activity of callus cultures of S. chirata in comparison to wild plant herbal drug samples.

Swertia L.: A comprehensive review of its genetic relationship, chemical compositions, pharmacological effects, toxicities, and applications.

Swertia L., as a commonly used ethnic medicine, is widely distributed in Sichuan, Yunnan, and Xizang in China. Moreover, the medicinal plants of Swertia L. have been widely used and constitute one of the most important sources of various traditional medicines in China due to their prominent activities. In this review, the information on the classification, distribution, genetic relationship, chemical composition, pharmacological effects, toxicities, and applications of the medicinal plants in Swertia L. was summarized based on the scientific literature. The results indicated that the medicinal plants of Swertia L. mainly contained chemical components including triterpenes, xanthones, and iridoids. These compounds exert pharmacological effects including ameliorating diseases related to the liver and gallbladder. They also exert antiviral and antibacterial effects and can alleviate the increase in blood glucose levels. Especially, prescriptions related to Swertia L. have been widely adopted in preclinical and clinical studies to protect against diseases affecting the liver and the gallbladder, including hepatitis, cirrhosis, and cholecystitis. In addition, it also discusses toxicity studies and future perspectives and provides a reference for their clinical development and utilization.

Secoiridoids from the traditional Chinese medicine Swertia pseudochinensis.

Detailed phytochemical investigation on the traditional Chinese medicine Swertia pseudochinensis Hara led to the isolation of ten undescribed secoiridoids and fifteen known analogs. Their structures were elucidated by extensive spectroscopic analysis (including 1D and 2D NMR, and HRESIMS). Selected isolates were assayed for their anti-inflammatory and antibacterial activities, and moderate anti-inflammatory activity via inhibiting the secretion of cytokines IL-6 and TNF-α in macrophages RAW264.7 induced by LPS were observed. Antibacterial activity against Staphylococcus aureus was not found at 100 µM.

Isolation and identification of novel phenolic and lignan glycosides from Swertia davidii Franch.

Phytochemical analyses of Swertia davidii Franch. extracts using column chromatography and semi-preparative HPLC were performed. Two novel phenolic glycosides named swertiosides A and B (compounds 1 and 2, respectively) were isolated and characterized. Four known phenolic glycosides were also extracted (compounds 3-6). The structural characteristics of these novel compounds were analyzed using 1D, 2D NMR, and HRMS. All six compounds have never been isolated from this particular plant species before this study. Subsequent assessment of bioactive properties suggested that compounds 1 and 2 exhibited moderate levels of cytotoxicity.

A rapid extraction process monitoring of Swertia mussotii Franch. With near infrared spectroscopy.

Swertia mussotii Franch. (SMF), a traditional Tibetan medicine, which has miraculous effect on treating hepatitis diseases. However, there is no research on its entire production process, and invisible production process has seriously hindered the SMF modern development. In this study, principal component analysis (PCA), subtractive spectroscopy, and two-dimensional correlation spectroscopy (2D-COS) were used to explain changes of characteristic groups in the extraction process. Four main characteristic peaks at 1884 nm, 1944 nm, 2246 nm and 2308 nm were identified to describe the changes of molecular structure information of total active components in SMF extraction process. In addition, multi critical quality attributes (CQAs) models were established by near-infrared spectroscopy (NIRS) combined with the total quantum statistical moment (TQSM). The coefficients of determination (R2eval and R2ival) were both greater than 0.99. The ratios of the standard deviation of validation to the standard error of the prediction (RPDe and RPDi) were greater than five. The quantitative model of AUCT could save time on primary data measurement by not requiring determination of indicator components compared with others. In conclusion, these results demonstrated that it was feasible to understand the SMF extraction process through AUCT and characteristic groups. These could realize the visual digital characterization and quality stability of the SMF extraction process.

Molecular recognition of bio-active triterpenoids from Swertia chirayita towards hepatitis Delta antigen: a mechanism through docking, dynamics simulation, Gibbs free energy landscape.

Medicinal plants the underpinning of indigenous herbal serve, are the possible source of key compounds for the development of new drugs. Hepatitis D, one of the most widespread infectious diseases associated with global public health issues. Therefore, we aim to screen natural compounds to find out potent inhibitor towards hepatitis delta antigen. Through ADMET investigation, we have screened twenty phytochemicals for this study. Additionally, using molecular docking, these phytochemicals were docked with the HDV protease which signifies the phytochemicals beta-amyrin, chiratenol, episwertenol and swertanone have a significant capability to bind with hepatitis D virus protein. The docking study was further accompanied by analyzes RMSD, RMSF, Rg, SASA, Hbond number, and principal component analysis through 100 ns MD simulations. Based on our principal component analysis, beta-amyrin, chiratenol, episwertenol and swertanone phytochemicals can be a potential drug candidates for inhibition of hepatitis D. The above observation is also supported by our Gibbs free energy landscape study. The potential therapeutic characteristics of the phytochemicals against hepatitis D inhibition offer additional support for the in vitro and in vivo studies in future.

Metabolite Profiling of Swertia cincta Extract in Rats and Pharmacokinetics Study of Three Bioactive Compounds Using UHPLC-MS/MS.

Swertia cincta, a plant of the genus Swertia in Gentianceae, has "heat-clearing" and detoxifying effects that normalize the gallbladder function in the treatment of jaundice. Although numerous studies on Swertia cincta have been performed, the absorption and pharmacokinetic behaviors remain unclear. In this study, the compounds of Swertia cincta in serum, bile, feces, and urine of rats were analyzed using a ultra-high-performance liquid chromatography-tandem mass spectrometry. A total of 9 prototype components and 48 metabolites were detected in biological samples. Furthermore, we determined the main components absorbed in the blood of Swertia cincta and established a method for simultaneously determining these components (sweroside, swertiamarin, and gentiopicroside) in positive ionization mode within 6 min. The quantitative method was successfully applied for the multiple-component pharmacokinetic study of Swertia cincta.

Three new constituents from the Tujia ethnomedicine Swertia punicea Hemsl.

Three new constituents: 1,5R-dihydroxy-3,8S-dimethoxy-5,6,7,8-tetrahydroxanthone (1), (3S,4R,16S,17R)-3,16,23-trihydroxyoleana-11,13(18)-dien-28-aldehyde-3-O-ß-D-glucopyranoside (2), and new natural product (S)-gentiandiol (3), along with 41 known compounds were isolated from Tujia ethnomedicine Shuihuanglian, namely, the whole plant of Swertia punicea. Structures of all these compounds were established through extensive spectroscopic techniques, namely 1D, 2D-NMR spectroscopy, HRESIMS analysis, and the absolute configuration of the new compounds was discerned by circular dichroism (CD) spectroscopy. Antioxidative effects of these compounds were evaluated by using the DPPH radical scavenging method, compounds 7, 9 and 14 showed antioxidant activities with IC50 values of 68.9, 50.8 and 48.2 µM, respectively.

Xanthones and Phenylpropanoids from the Whole Herb of Swertia pseudochinensis and Their Anti-Inflammatory Activity.

An undescribed xanthone dimer, 1,3,5,8-tetrahydroxy-7-(1',5',8'-trihydroxy-3'-methoxy-2'-xanthonyl)xanthone (1) was separated together with eleven known compounds (2-12) from the dried whole herb of Swertia pseudochinensis. It was the first time that the compounds 8-12 were isolated from the Swertia genus. The structure of compound 1 was illuminated based on chemical evidence and spectral data analysis (UV, 1D and 2D-NMR, HR-ESI-MS). Moreover, the inhibitory effects of all compounds on NO production in LPS-induced RAW 264.7 cells were tested, compounds 8, 9, 10, 11 and 12 showing significant inhibition. The IC50 value of compound 12 was 3.05±1.10 µM. Using target screening and molecular docking, we hypothesized that compound 12 may bind neutrophil elastase to exert its anti-inflammatory effects.

Swertia purpurascens Wall ethanolic extract mitigates hepatic fibrosis and restores hepatic hepcidin levels via inhibition of TGFß/SMAD/NFκB signaling in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Swertia purpurascens Wall belongs to a well-known genus in traditional systems of medicine worldwide. In folklore, it is used to treat various ailments, including hepatic disorders, as an alternative to the endangered species Swertia chirayita. However, the therapeutic potential of Swertia purpurascens Wall against hepatic fibrosis has not been validated yet. AIM OF THE STUDY: The present study was planned to evaluate the efficacy of the Swertia purpurascens Wall extract (SPE) against hepatic fibrosis and elucidate the underlying mechanism of action. MATERIALS AND METHODS: The metabolite profiling of the SPE was done using UHPLC-QTOF-MS/MS. The acute oral toxicity study of SPE at 2 g/kg BW dose was done in rats. Further, the liver fibrosis was induced by the CCl4 intoxication, and the efficacy of SPE at three doses (100, 200 and 400 mg/kg BW) was evaluated by studying biochemical parameters, histopathology, immunohistochemistry, qRT-PCR, western blotting and in silico analysis. RESULTS: UHPLC-QTOF-MS/MS analysis revealed the presence of a total of 23 compounds in SPE. Acute oral toxicity study of SPE at 2 g/kg BW showed no harmful effects in rats. Further, the liver fibrosis was induced by the CCl4 administration, and the efficacy of SPE was evaluated at three doses (100, 200 and 400 mg/kg BW). SPE treatment significantly improved the body weight gain, the relative liver weight, serum liver injury markers and endogenous antioxidant enzyme levels in the CCl4-treated rats. SPE also recovered the altered liver histology and effectively reduced the fibrotic tissue deposition in the hepatic parenchyma. Further, SPE significantly inhibited the fibrotic (TGFß, αSMA, SMADs and Col1A), proinflammatory markers (NFκB, TNFα and IL1ß) and apoptosis in the liver tissue. Interestingly, SPE treatment also restored the altered hepcidin levels in the liver tissue. In silico study revealed the potential of various metabolites as drug candidates and their interaction with target proteins. CONCLUSION: Altogether, SPE showed its therapeutic potential against CCl4-induced hepatic fibrosis by restoring the hepatic hepcidin levels and inhibiting TGFß/SMAD/NFκB signaling in rats.

Isolation, optimized extraction, and ultra-high performance liquid chromatography with photodiode array method for quantitative analysis of chiratol in Swertia paniculata.

The global natural product-based industry is growing fast with the introduction of new phytochemicals and herbal extract products from different geographical regions. Swertia paniculata is a well-known plant with medicinal properties; however, the quality control for its major phytochemical constituents from the Himalayan geographical region is nevertheless reported. Therefore, the first objective of this investigation was to characterize and optimize the extraction process while the second objective was to validate a quantitative analytical method for chiratol from S. paniculata herbal extract. The chiratol was characterized with spectral analysis. The optimum extraction condition for the highest yield of metabolite was realized in chloroform as a solvent system under ultrasonication. The ultra-high performance liquid chromatography coupled with photodiode array detection method for analytical quantification was validated for specificity, linearity, limits of detection, limits of quantification, precision, repeatability, recovery, and robustness using Eclipse Plus C18 column (100 mm × 4.6 mm × 3.5 µm id). The gradient elution of water/acetonitrile as mobile phase was used at a flow rate of 0.5 ml/min. The recovery percentage was very satisfactory with values within specification. The robustness parameters showed no substantial influence of evaluated parameters by the Youden test. The developed method was ascertained to be appropriate for the proposed purpose.

Optimization of harvest and extraction factors by full factorial design for the improved yield of C-glucosyl xanthone mangiferin from Swertia chirata.

Swertia chirata Buch.-Ham. ex C.B. Clarke is an important medicinal plant used in various herbal formulations as it shows significant biological activities such as hepatoprotective, hypoglycemic, anti-inflammatory, antimalarial, antioxidant and anti-parkinson. C-glucosyl xanthone glycoside (mangiferin) is known as bio-marker compound of genus Swertia L. Development of efficient extraction methods of C-glucosyl xanthone mangiferin from Swertia chirata was attempted by optimizing the pre-harvest, post-harvest and extraction techniques by full factorial design. Firstly, a full factorial design was implemented to evaluate the single and interactive effects of pre-harvest (growth stage and plant part), post-harvest (drying condition and storage periods) followed by selection of best extraction technique such as heat reflux extraction (HRE), microwave assisted extraction (MAE) and ultrasound assistant extraction (UAE) at different solvent types on mangiferin yield. HPTLC and HPLC techniques were used for the determination of mangiferin content in extracts generated from different plant samples. In addition, anti-oxidant and anti-diabetic properties were determined by using DPPH assay and percentage inhibition of α­amylase enzyme. Substantial variation of mangiferin yield, ranged from 1.46 to 4.86% was observed, depending on the growth stage, plant part, drying condition, storage periods and extraction method. Results showed that drying of the leaves of Swertia chirata in the shade harvested at budding stage and stored for not more than 1 month was recommended for obtaining a higher mangiferin yield. Among different extraction techniques, MAE and UAE in 50% aqueous ethanol solvent were found to be efficient and cost-effective with better yield of mangiferin (4.82% and 4.86%, respectively) as compared to HRE (4.14%). Highest DPPH activity and percentage inhibition of α­amylase was observed in the aqueous ethanol extract of S. chirata leaves harvested at bud-stage of plant followed by flowering stage. The study shows that optimization of various factors by full factorial design was found to be an effective procedure to improve mangiferin yield from Swertia chirata and can be used for extraction of mangiferin.

Iridoid compounds from the aerial parts of Swertia mussotii Franch. with cytotoxic activity.

One new secoiridoid compound swertiamarin B (1), along with a known compound lytanthosalin (2), were isolated from ethanol extract of the aerial parts of Swertia mussotii. Their structures were elucidated by the detailed analysis of comprehensive spectroscopic data. All compounds were first isolated from the Swertia genus. Their antitumor activities were evaluated for four human tumor cell lines (HCT-116, HepG2, MGC-803 and A549). Compounds 1 and 2 showed excellent cytotoxic activities toward the MGC-803 cell lines with IC50 values 3.61 and 12.04 µM, respectively.

Chemical profile of Swertia mussotii Franch and its potential targets against liver fibrosis revealed by cross-platform metabolomics.

ETHNOPHARMACOLOGICAL RELEVANCE: Swertia mussotii Franch (SMF) is a well-known Tibetan medicine for the treatment of liver disease in China. However, the chemical profile and molecular mechanism of SMF against hepatic fibrosis are not yet well explored. AIM OF THE STUDY: This work aimed to elucidate the chemical profile of SMF and investigate the action mechanisms of SMF against carbon tetrachloride (CCl4)-induced hepatic fibrosis. MATERIALS AND METHODS: Ultra performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOFMS) and UNIFI platform was firstly employed to reveal the chemical profile of SMF. Cross-platform serum metabolomics based on gas chromatography/liquid chromatography-mass spectrometry were performed to characterize the metabolic fluctuations associated with CCl4-induced hepatic fibrosis in mice and elucidate the underlying mechanisms of SMF. Western blotting was further applied to validate the key metabolic pathways. RESULTS: A total of 31 compounds were identified or tentatively characterized from SMF. Twenty-seven differential metabolites were identified related with CCl4-induced liver fibrosis, and SMF could significantly reverse the abnormalities of seventeen metabolites. The SMF-reversed metabolites were involved in arachidonic acid metabolism, glycine, serine and threonine metabolism, tyrosine metabolism, arginine and proline metabolism, primary bile acid biosynthesis, glycerophospholipid metabolism and TCA cycle. The results of western blotting analysis showed that SMF could alleviate liver fibrosis by increasing the levels of CYP7A1, CYP27A1 and CYP8B1 and decreasing the level of LPCAT1 to regulate the metabolic disorders of primary bile acid biosynthesis and glycerophospholipid. CONCLUSION: It could be concluded that primary bile acid biosynthesis and glycerophospholipid metabolism were the two important target pathways for SMF-against liver fibrosis, which provided the theoretical foundation for its clinical use.

Sweritranslactone D, a hepatoprotective novel secoiridoid dimer with tetracyclic lactone skeleton from heat-transformed swertiamarin.

Swertia mileensis, known as Qing-Ye-Dan (QYD), has been documented in Chinese Pharmacopoeia to cure hepatitis. Interestingly, its announced main active component, swertiamarin, could not be detected in the decoction, which indicated that the efficacy of QYD might be attributed to heat-transformed products of swertiamarin (HTPS). Further investigation on HTPS led to the isolation of sweritranslactone D (1), a novel secoiridoid dimer possessing a tetracyclic lactone skeleton, with better hepatoprotective activity than N-acetyl-L-cysteine in vitro.

Exploring the Mechanism of Swertia mussotii Franch. for Hepatoprotective Effects with iTRAQ LC-MS/MS.

BACKGROUND AND OBJECTIVE: A Tibetan traditional herb named Swertia mussotii Franch., also called "Zangyinchen" by the local people of the Qinghai-Tibet area, has been used to protect the liver from injury for many years. However, the curative effect and molecular mechanism of the herb have not been demonstrated clearly. MATERIALS AND METHODS: In our study, serum alanine aminotransferase, aspartate aminotransferase, and total bilirubin levels were examined after S. mussotii Franch. treatment of the acute liver injury on the carbon tetrachloride-induced rat model. Then, proteome analysis was conducted to explore the potential mechanism of SMT for hepatoprotective effects after iTRAQLC-MS/MS analysis (isobaric tag for relative and absolute quantification-liquid chromatography-mass spectrometer with tandem mass spectrometry). RESULTS: Serum results showed that alanine aminotransferase, aspartate aminotransferase, and total bilirubin levels of rats with acute liver injury were all improved with SMT treatment. Moreover, proteome analysis suggested that, with S. mussotii Franch. treatment, the levels of lipid catabolic process and lipid homeostasis were all enhanced. Furthermore, the results of protein-protein interaction (PPI) analysis illustrated that the proteins assembled in PPI networks were found to be significantly enriched in response to lipid, negative regulation of lipase activity, and in response to lipopolysaccharides. Furthermore, the down-regulated MRP14 and MRP8 proteins were found to be involved in the lipid metabolism, which may indicate the mechanism of SMT for the protection of the liver from ALI induced by carbon tetrachloride. CONCLUSION: SMT herb could play a role in hepatoprotection and alleviating the acute liver injury by impacting the lipid metabolism associated with the biological process.

Anti-Metastatic Potential of a Novel Xanthone Sourced by Swertia chirata Against In Vivo and In Vitro Breast Adenocarcinoma Frameworks.

BACKGROUND: The Anticancer property of Swertia chirata has been well established. It forms a rich source of compounds to which its anticancer property can be attributed, among the compounds found in S. chirata xanthones form an important group. Among the most abundant xanthones found in S. chirata, 1,5,8-trihydroxy-3-methoxy xanthone (TMX) was found to be most effective. As metastasis is the underlying cause of most cancer-related deaths, in this study, we evaluated the anti-metastatic potential of TMX against adenocarcinoma both in vivo and in vitro. MATERIALS AND METHODS: In vivo anti-metastatic potential was proved by histological evidence of different organs, giemsa staining of bone marrow, subcutaneous re-injection of the aberrant bone marrow cells into the right flank of the mice to observe the formation of tumors and analyzing the markers related to metastasis by immunohistochemistry (IHC) and western blot. In vitro validation of anti-metastatic potential was carried out against human breast adenocarcinoma cell line MCF-7 by primarily analyzing the migratory property of cells through scratch wound healing assay and the ability of cells to form colonies. The re-validation part was performed by western blot of markers related to metastasis and real-time analysis of EMT related markers. RESULTS: In vivo, TMX treatment restricted metastasis of EAC induced solid tumor to liver, lung, bone marrow, and validation of this finding was achieved by down regulation of metastatic and EMT markers.  In vitro, TMX treatment restricted migratory and colony forming ability of MCF-7 cells by down regulating metastatic and EMT markers. CONCLUSION: It was proved from our study that TMX treatment successfully reduced the metastatic potential of EAC induced solid tumor, with in vitro validation TMX on the MCF-7 cell line.