Silybin

Development of an HPLC-MS/MS Method for the Determination of Silybin in Human Plasma, Urine and Breast Tissue.[Pubmed:32599946]

Molecules. 2020 Jun 24;25(12). pii: molecules25122918.

Silybin is a flavonolignan extracted from Silybum marianum with chemopreventive activity against various cancers, including breast. This study was designed to develop an HPLC-MS/MS method for the determination of Silybin in human plasma, urine and breast tissue in early breast cancer patients undergoing Siliphos((R)) supplementation, an oral Silybin-phosphatidylcholine complex. The determination of Silybin was carried out by liquid-liquid extraction with methyl-tert-butyl ether (MTBE); total Silybin concentration was determined by treating the samples with beta-glucuronidase, while for the determination of free Silybin, the hydrolytic step was omitted. Naringenin and naproxen were selected as internal standards. The detection of the analyte was carried out by mass spectrometry and by chromatography. The HPLC-MS/MS method was evaluated in terms of selectivity, linearity, limit of quantification, precision and accuracy, and carryover. The method proved to be selective, linear, precise and accurate for the determination of Silybin. To the best of our knowledge, this presents the first analytical method with the capacity to quantify the major bioactive components of milk thistle in three different biological matrices with a lower limit of quantification of 0.5 ng/mL for plasma. Silybin phosphatidylcholine, taken orally, can deliver high blood concentrations of Silybin, which selectively accumulates in breast tumor tissue.

Artemlavanins A and B from Artemisia lavandulaefolia and Their Cytotoxicity Against Hepatic Stellate Cell Line LX2.[Pubmed:32583278]

Nat Prod Bioprospect. 2020 Jun 24. pii: 10.1007/s13659-020-00254-0.

Two new sesquiterpenoids, artemlavanins A (1) and B (3), together with fifteen known compounds (2 and 4-17) were isolated from the EtOH extract of Artemisia lavandulaefolia. The structures of new compounds were elucidated by extensive spectroscopic analyses (HRESIMS, 1D and 2D NMR) and ECD calculations. Compound 1 was a sesquiterpenoid lactone possessing a rearranged eudesmane skeleton; compounds 2-5, 6-8, 9 and 10-12 belonged to the eudesmane, guaiane, oppositane and farnesane sesquiterpenoids, respectively; compounds 13-17 were the phenyl derivatives with a 4-hydroxyacetophenone moiety. Twelve compounds (1-3, 5-7, 10-12, 14, 15 and 17) displayed cytotoxicity against hepatic stellate cell line LX2 (HSC-LX2) with IC50 values ranging from 35.1 to 370.3 muM. Compounds 2, 7, 10-12 and 17 exhibited the stronger cytotoxicity than Silybin (IC50, 169.6 muM) with IC50 values of 82.1, 35.1, 95.0, 83.8, 81.6 and 90.1 muM. Compound 7 as the most active one showed significant inhibition on the deposition of human collagen type I (Col I), human hyaluronic acid (HA) and human laminin (HL) with IC50 values of 10.7, 24.5 and 13.3 muM.

Multidrug Resistance Modulation Activity of Silybin Derivatives and Their Anti-inflammatory Potential.[Pubmed:32466263]

Antioxidants (Basel). 2020 May 25;9(5). pii: antiox9050455.

Silybin is considered to be the main biologically active component of silymarin. Its oxidized derivative 2,3-dehydroSilybin typically occurs in silymarin in small, but non-negligible amounts (up to 3%). Here, we investigated in detail complex biological activities of Silybin and 2,3-dehydroSilybin optical isomers. Antioxidant activities of pure stereomers A and B of Silybin and 2,3-dehydroSilybin, as well as their racemic mixtures, were investigated by using oxygen radical absorption capacity (ORAC) and cellular antioxidant activity (CAA) assay. All substances efficiently reduced nitric oxide production and cytokines (TNF-alpha, IL-6) release in a dose-dependent manner. Multidrug resistance (MDR) modulating potential was evaluated as inhibition of P-glycoprotein (P-gp) ATPase activity and regulation of ATP-binding cassette (ABC) protein expression. All the tested compounds showed strong dose-dependent inhibition of P-gp pump. Moreover, 2,3-dehydroSilybin A (30 microM) displayed the strongest sensitization of doxorubicin-resistant ovarian carcinoma. Despite these significant effects, Silybin B was the only compound acting directly upon P-gp in vitro and also downregulating the expression of respective MDR genes. This compound altered the expression of P-glycoprotein (P-gp, ABCB1), multidrug resistance-associated protein 1 (MRP1, ABCC1) and breast cancer resistance protein (BCRP, ABCG2). 2,3-DehydroSilybin AB exhibited the most effective inhibition of acetylcholinesterase activity. We can clearly postulate that Silybin derivatives could serve well as modulators of a cancer drug-resistant phenotype.

Flavolignans from Silymarin as Nrf2 Bioactivators and Their Therapeutic Applications.[Pubmed:32423098]

Biomedicines. 2020 May 14;8(5). pii: biomedicines8050122.

Silymarin (SM) is a mixture of flavolignans extracted from the seeds of species derived from Silybum marianum, commonly known as milk thistle or St. Mary'sthistle. These species have been widely used in the treatment of liver disorders in traditional medicine since ancient times. Several properties had been attributed to the major SM flavolignans components, identified as Silybin, isoSilybin, silychristin, isosilychristin, and silydianin. Previous research reported antioxidant and protective activities, which are probably related to the activation of the nuclear factor erythroid 2 (NFE2)-related factor 2 (Nrf2), known as a master regulator of the cytoprotector response. Nrf2 is a redox-sensitive nuclear transcription factor able to induce the downstream-associated genes. The disruption of Nrf2 signaling has been associated with different pathological conditions. Some identified phytochemicals from SM had shown to participate in the Nrf2 signaling pathway; in particular, they have been suggested as activators that disrupt interactions in the Keap1-Nrf2 system, but also as antioxidants or with additional actions regarding Nrf2 regulation. Thus, the study of these molecules makes them appear attractive as novel targets for the treatment or prevention of several diseases.

Synthesis of bio-mediated silver nanoparticles from Silybum marianum and their biological and clinical activities.[Pubmed:32409047]

Mater Sci Eng C Mater Biol Appl. 2020 Jul;112:110889.

The purpose of current study was green synthesis of silver nanoparticles (AgNPs) from seeds and wild Silybum plants in comparison with their respective extracts followed by characterization and biological potency. The biologically synthesized AgNPs were subjected to characterization using techniques like XRD, FTIR, TEM, HPLC and SPE. Highly crystalline and stable NPs were obtained using Silybum wild plant (NP1) and seeds (NP3) with size range between 18.12 and 13.20 nm respectively. The synthesized NPs and their respective extracts revealed a vast range of biological applications showing antibacterial, antioxidant, anti-inflammatory, cytotoxic and anti-aging potencies. The highest antioxidant activity (478.23 +/- 1.9 muM, 176.91 +/- 1.3 muM, 83.5 +/- 1.6% mugAAE/mg, 156.32 +/- 0.6 mugAAE/mg) for ABTS, FRAP, FRSA, TRP respectively was shown by seed extract (NP4) followed by highest value of (117.35 +/- 0.9 mugAAE/mg) for TAC by wild extract (NP2). The highest antifungal activity (13 mm +/- 0.76) against Candida albicans was shown by NP3 while antibacterial activity of (6 mm against Klebsiella pneumonia) was shown by NP3 and NP4. The highest anti-inflammatory activity (38.56 +/- 1.29 against COX1) was shown by NP2. Similarly, the high value of (48.89 +/- 1.34 against Pentosidine-Like AGEs) was shown by NP4. Also, the high anti-diabetic activity (38.74 +/- 1.09 against alpha-amylase) was shown by NP4. The extracts and the synthesized NPs have shown activity against hepato-cellular carcinoma (HepG2) human cells. The HPLC analysis revealed that the highest value of silymarin component (Silybin B 2289 mg/g DW) was found for NP4. Silydianin is responsible for capping. Among the green synthesized AgNPs and the extracts used, the effect of NP4 was most promising for further use.

The Bisphenol A Induced Oxidative Stress in Non-Alcoholic Fatty Liver Disease Male Patients: A Clinical Strategy to Antagonize the Progression of the Disease.[Pubmed:32408667]

Int J Environ Res Public Health. 2020 May 12;17(10). pii: ijerph17103369.

: Introduction: Bisphenol A (BPA) exposure has been correlated to non-alcoholic fatty liver disease (NAFLD) development and progression. We investigated, in a clinical model, the effects of the administration of 303 mg of Silybin phospholipids complex, 10 g of vitamin D, and 15 mg of vitamin E (RealSIL, 100D, IBI-Lorenzini, Aprilia, Italy) in male NAFLD patients exposed to BPA on metabolic, hormonal, and oxidative stress-related parameters. METHODS: We enrolled 32 male patients with histologic diagnosis of NAFLD and treated them with Realsil 100D twice a day for six months. We performed at baseline clinical, biochemical, and food consumption assessments as well as the evaluation of physical exercise, thiobarbituric acid reactive substances (TBARS), plasmatic and urinary BPA and estrogen levels. The results obtained were compared with those of healthy control subjects and, in the NAFLD group, between baseline and the end of treatment. RESULTS: A direct proportionality between TBARS levels and BPA exposure was shown (p < 0.0001). The therapy determined a reduction of TBARS levels (p = 0.011), an improvement of alanine aminotransferase, aspartate aminotransferase, insulinemia, homeostatic model assessment insulin resistance, C reactive protein, tumor necrosis factor alpha (p < 0.05), an increase of conjugated BPA urine amount, and a reduction of its free form (p < 0.0001; p = 0.0002). Moreover, the therapy caused an increase of plasmatic levels of the native form of estrogens (p = 0.03). CONCLUSIONS: We highlighted the potential role of BPA in estrogen oxidation and oxidative stress in NAFLD patients. The use of Realsil 100D could contribute to fast BPA detoxification and to improve cellular antioxidant power, defending the integrity of biological estrogen-dependent pathways.

Silybin enhances the blood concentration of brexpiprazole in rats by downregulating expression of CYP3A4 and CYP2D6.[Pubmed:32393428]

Pharmazie. 2020 May 1;75(5):198-200.

In this study, we investigated the effect of Silybin on the pharmacokinetics of brexpiprazole and the underlying mechanism in rats. Two groups of animals received Silybin at different doses (50 mg/kg, 25 mg/kg) for 2 weeks, while another group was given vehicle alone. After that, rats were intragastrically administrated with 2 mg/kg brexpiprazole. Then, the tail blood and liver tissues were collected from each rat at different time points. Brexpiprazole in serum was determined by an established UPLC-MS/MS assay. Finally, pharmacokinetic parameters of animals in each group were figured out. The results show that Silybin remarkably changed the pharmacokinetic properties of brexpiprazole, especially at the highest dose. AUC and Cmax in the combination group with 50 mg/kg Silybin were enhanced approximately 4 times as much as after a single dose of brexpiprazole, p < 0.05. Meanwhile, total liver protein of each sample was extracted, and was subjected to immunoblotting assay for probing CYP3A4 and CYP2D6. Therein CYP3A4 was significantly downregulated compared to the control group. Overall, Silybin can increase blood concentration of brexpiprazole in rat by downregulating its main metabolic enzyme CYP3A4. Therefore, the maintenance dose of brexpiprazole should be decreased while co-treating with Silybin.

Silymarin and Cancer: A Dual Strategy in Both in Chemoprevention and Chemosensitivity.[Pubmed:32344919]

Molecules. 2020 Apr 25;25(9). pii: molecules25092009.

Silymarin extracted from milk thistle consisting of flavonolignan Silybin has shown chemopreventive and chemosensitizing activity against various cancers. The present review summarizes the current knowledge on the potential targets of silymarin against various cancers. Silymarin may play on the system of xenobiotics, metabolizing enzymes (phase I and phase II) to protect normal cells against various toxic molecules or to protect against deleterious effects of chemotherapeutic agents on normal cells. Furthermore, silymarin and its main bioactive compounds inhibit organic anion transporters (OAT) and ATP-binding cassettes (ABC) transporters, thus contributing to counteracting potential chemoresistance. Silymarin and its derivatives play a double role, namely, limiting the progression of cancer cells through different phases of the cycle-thus forcing them to evolve towards a process of cell death-and accumulating cancer cells in a phase of the cell cycle-thus making it possible to target a greater number of tumor cells with a specific anticancer agent. Silymarin exerts a chemopreventive effect by inducing intrinsic and extrinsic pathways and reactivating cell death pathways by modulation of the ratio of proapoptotic/antiapoptotic proteins and synergizing with agonists of death domains receptors. In summary, we highlight how silymarin may act as a chemopreventive agent and a chemosensitizer through multiple pathways.

Evaluation of Mogroside V as a Promising Carrier in Drug Delivery: Improving the Bioavailability and Liver Distribution of Silybin.[Pubmed:32337654]

AAPS PharmSciTech. 2020 Apr 26;21(4):123.

The objective of this work was to investigate the capacity of mogroside V (MOG-V), a food additive, as a novel carrier to improve the bioavailability and liver distribution of Silybin (SLY). Solid dispersion particles (SDPs) of SLY/MOG-V were prepared utilizing the solvent evaporation method. The physicochemical characterizations of SDPs were evaluated by using dynamic light scattering (DLS), differential scanning calorimetry (DSC), and powder X-ray diffraction (PXRD) measurements. DLS results demonstrated the formation of nanoparticles (206 nm) of SDPs in water. DSC and PXRD analysis revealed that SLY was in amorphous form or molecularly dispersed in SDPs. SDPs also exhibited a major increase in both dissolution rate and saturation solubility, as evidenced by a 1931-fold improvement (2201 mug/mL) in solubility compared with pure SLY (1.14 mug/mL). The pharmacokinetic study in rats showed that oral absorption of SLY/MOG-V SDPs was dramatically increased. The mean value of AUC until 12 h for SLY/MOG-V SDPs (27,481 ng.min/mL) was 24.5-fold higher than that of pure SLY (1122 ng.min/mL). In vivo tissue distribution experiment in mice confirmed that the major distribution tissue was changed from lungs to liver after SLY was loaded into MOG-V. In addition, even orally administrated to mice at a high dose (4.2 g/kg), MOG-V exhibited no undesirable effect on the plasma glucose concentrations. Thus, MOG-V may have the applicability to serve as an ideal excipient for solubilization or as a novel liver targeting carrier for the delivery of SLY.

Phytochemicals protect L02 cells against hepatotoxicity induced by emodin via the Nrf2 signaling pathway.[Pubmed:32153769]

Toxicol Res (Camb). 2019 Nov 20;8(6):1028-1034.

Dihydromyricetin (DMY), hyperoside and Silybin are phytochemicals that belong to a class called flavonoids, and they have been used in liver protection pharmaceutical preparations, but the specific mechanism of these chemicals is still unclarified. This study aims to investigate the hepatoprotective effects and potential mechanism of these phytochemicals. The immortalized human hepatocyte cell line L02 was treated with 200 muM emodin for 48 h, and this was used as a hepatocyte injury model. The L02 cells were treated with both 200 muM emodin and different concentrations of DMY/hyperoside/Silybin for 48 h to investigate the protective effects of these phytochemicals. The CCK-8 assay was used to detect cell viability. RT-qPCR and western blotting were performed to examine the mRNA and protein expression, respectively, of the classic bile acid synthetic pathway gene CYP7A1, the bile acid efflux transporter bile salt export pump (BSEP), the nuclear factor erythroid-2-related factor 2 (Nrf2) and the drug processing gene CYP1A2. DMY, hyperoside and Silybin prevented the impairment of cell viability that was caused by emodin-induced hepatotoxicity in a dose-dependent manner, and at a low concentration (10 muM), the protective effect followed the order hyperoside > DMY > Silybin, while at a high concentration (160 muM), the protective effect followed the order DMY > hyperoside > Silybin. These phytochemicals reduced the expression of CYP7A1 at both the mRNA and protein levels. BSEP was not influenced by the phytochemical intervention. When 200 muM emodin was used for 48 h with the addition of the phytochemicals at 200 muM, the nuclear protein expression of Nrf2 significantly increased and CYP1A2 expression decreased. DMY, hyperoside and Silybin prevented the hepatotoxicity induced by emodin in the L02 cells, potentially, via the Nrf2 signaling pathway.

Co-delivery of silybin and paclitaxel by dextran-based nanoparticles for effective anti-tumor treatment through chemotherapy sensitization and microenvironment modulation.[Pubmed:32044390]

J Control Release. 2020 May 10;321:198-210.

Modulation of tumor microenvironment (TME) has been indicated as an approach to improve efficacy of cancer therapy. Here, we proposed a nano co-delivery based combination therapy of paclitaxel (PTX) and Silybin (SB) which can employ the synergistic effects through chemotherapy sensitization and microenvironment modulation. A dextran-based amphiphilic polymer (Dex-DOCA) was successfully developed for in vivo co-delivery and thus "synchronizing" the biodistribution, transport and release of PTX and SB. Resultantly, Dex-DOCA exhibited an excellent encapsulating efficiency for both PTX and SB with adjustable loading ratio for an optimal synergistic antitumor activity. Moreover, the co-loaded nanoparticles efficiently discharged the two drugs at the prospective dosage ratio specifically in acid endo/lysosome mimic environments. The results of in vitro cytotoxicity and cell apoptosis assays further confirmed the SB sensitized PTX potency. Finally, in vivo investigation demonstrated that the co-loaded nanoparticles could effectively accumulate in tumor sites by passive targeting, and inhibit tumor growth through an enhanced intratumoral penetration (resulted from stromal components eradication and tumor vessels normalization associated TME modulation), as well as a sensitization effect of SB on PTX cytotoxic chemotherapy.

Green production of silybin and isosilybin by merging metabolic engineering approaches and enzymatic catalysis.[Pubmed:32004707]

Metab Eng. 2020 May;59:44-52.

Silymarin extracted from milk thistle seeds, is used for treating hepatic diseases. Silybin and isoSilybin are its main components, and synthesized from coupling of taxifolin and coniferyl alcohol. Here, the biosynthetic pathways of taxifolin and coniferyl alcohol were reconstructed in Saccharomyces cerevisiae for the first time. To alleviate substantial burden caused by a great deal of genetic manipulation, expression of the enzymes (e.g. ZWF1, TYR1 and ARO8) playing multiple roles in the relevant biosynthetic pathways was selectively optimized. The strain YT1035 overexpressing seven heterologous enzymes and five native enzymes and the strain YC1053 overexpressing seven heterologous enzymes and four native enzymes, respectively produce 336.8 mg/L taxifolin and 201.1 mg/L coniferyl alcohol. Silybin and isoSilybin are synthesized from taxifolin and coniferyl alcohol under catalysis of APX1t (the truncated milk thistle peroxidase), with a yield of 62.5%. This study demonstrates an approach for producing Silybin and isoSilybin from glucose for the first time.

Flavonolignans: One Step Further in the Broad-Spectrum Approach of Cancer.[Pubmed:31976848]

Anticancer Agents Med Chem. 2020 Jan 23. pii: ACAMC-EPUB-103896.

BACKGROUND: The small chemical class of flavonolignans encompasses unique hybrid molecules with versatile biological activities. Their anticancer effects have received considerable attention, and a large body of supporting evidence has accumulated. Moreover, their ability to interact with proteins involved in drug resistance, and to enhance the effects of conventional chemotherapeutics in decreasing cell viability make them influential partners in addressing cancer. OBJECTIVE: The review provides an outline of the various ways in which flavonolignans advance the combat against cancer. While the main focus falls on flavonolignans from milk thistle, attention is drawn to the yet underexplored potential of less known flavonolignan subgroups derived from isoflavonoids and aurones. METHODS: Proceeding from the presentation of natural flavonolignan subtypes and their occurrence, the present work reviews these compounds with regard to their molecular targets in cancer, anti-angiogenetic effects, synergistic efficacy in conjunction with anticancer agents, reversal of drug resistance, and importance in overcoming the side effects of anticancer therapy. Recent advances in the endeavor to improve flavonolignan bioavailability in cancer are also presented. CONCLUSIONS: Significant progress has been achieved in detailing the molecular mechanisms of Silybin and its congeners in experimental models of cancer. The availability of novel formulations with improved bioavailability, and data from phase I clinical trials in cancer patients provide an encouraging basis for more extensive trials aimed at evaluating the benefits of Silybum flavonolignans in cancer management. On the other hand, further research on the antitumor efficacy of isoflavonolignans and other subtypes of flavonolignans should be pursued.