Effect of erythrocytes, plasma proteins, and lysosomes on systemic exposure to tetrandrine and fangchinoline / 中国临床药理学与治疗学

AlM: The Chinese medicinal herb Hanfangji is dried roots of Stephania tetrandra S. Moore (Family, Menispermaceae). Tetrandrine and fangchinoline are two major constituents of Hanfangji and these bisbenzyltetrahydroisoquinoline alkaloids possess anti - cancer and other pharmacological activities. To facilitate further pharmacodynamic investigation of these compounds, a pharmacokinetic investigation was performed in rats and in vitro. METHODS: Pharmacokinetics of tetrandrine and fangchinoline were characterized in rats p.o. or i.v. dosing an aqueous extract of Hanfangji or the individual compound. Unbound levels of systemic exposure to these two alkaloids were assessed using in vitro studies of plasma protein binding, blood-plasma partition, and lysosomal trapping. All the study samples were analyzed by liquid chromatography/mass spectrometry.RESULTS: We found two pharmacokinetic features of tetrandrine and fangchinoline. First, the two compounds had blood levels of systemic exposure substantially higher than the respective plasma levels of systemic exposure. Second, the two compounds exhibited significantly higher systemic exposure levels after p.o. dosing an aqueous extract of Hanfangji than the respective exposure levels after p.o. dosing the individual compound, at the same compound dose levels and under the same conditions for analytical measurement and the same conditions for animal study. Unbound fractions of tetrandrine and fangchinoline in rat plasma were 2%-5% and the concentrations of the alkaloids in rat erythrocytes were 5-times higher than those in rat plasma. Lysosomal inhibitor could block their trapping in lysosomes and significantly reduce their concentrations in HEK-293 cells. CONCLUSlON: The following pharmacokinetic aspects should be noted in pharmacodynamic investigation of tetrandrine and fangchinoline: extensive binding with plasma proteins, extensive binding with erythrocytes, and trapping by lysosomes of tissue cells substantially reduce the levels of unbound tetrandrine and fangchinoline in the systemic circulation.

Effect of fangchinoline on oxidant status in male albino rats with streptozotocin-induced diabetes

BACKGROUND Diabetes is a metabolic disorder caused by defects in insulin production and activity. During disease progression, changes in lipid peroxidation cause structural modifications via production of free radicals. Fangchinoline is a well-known alkaloid present in Stephaniae tetrandrine S. Moore, which has demonstrated antioxidant, anticancer, and anti-inflammatory activities. RESULTS The present study analyzed the anti-diabetic and antioxidant effects of fangchinoline in male rats with streptozotocin-induced diabetes. Rats were divided into the following groups: normal control, diabetic, diabetic + fangchinoline 100 mg/kg, diabetic + fangchinoline 200 mg/kg and diabetic + glibencla mide 600 mg/kg. The treatment was administered orally for 45 consecutive days. Lipid peroxidation was substantially increased by >50% in the serum, as well as the liver, kidney, and heart tissues of diabetic rats. However, fangchinoline supplementation significantly reduced lipid peroxidation to near normal levels. Reactive oxygen species levels were substantially increased by >500% in the serum, as well as the liver, kidney, and heart tissues of diabetic rats. Fangchinoline supplementation reduced reactive oxygen species to near normal levels. Fangchinoline supplementation significantly improved superoxide dismutase, glutathione peroxidase, catalase, and reduced glutathione levels in diabetic rats. Total hexoses, sialic acid, hexosamines, and fucose were increased in diabetic rats, whereas fangchinoline supplementation significantly reduced these total hexoses, sialic acid, hexosamines, and fucose to near normal levels CONCLUSIONS Supplementation with fangchinoline led to significant attenuation of the levels of lipid peroxidation, ROS, and glycoprotein components such as total hexoses, hexosamines, sialic acid, and fucose, while improving antioxidant marker levels

An overview on the chemistry, pharmacology and anticancer properties of tetrandrine and fangchinoline (alkaloids) from Stephania tetrandra roots / 中西医结合学报

Tetrandrine (TET) and fangchinoline (FAN) are dominant bisbenzylisoquinoline (BBIQ) alkaloids from the roots of Stephania tetrandra of the family Menispermaceae. BBIQ alkaloids comprise two benzylisoquinoline units linked by oxygen bridges. The molecular structures of TET and FAN are exactly the same, except that TET has a methoxy (-OCH

Screening of HDAC3/8 Small Molecule Inhibitors from Traditional Chinese Medicine Based on Molecular Docking / 中国实验方剂学杂志

Objective::To discover a small molecule active ingredient of traditional Chinese medicine (TCM) with the inhibitory activity of histone deacetylase (HDAC) 3/8. Method::The molecular docking technique was performed by AutoDock 4.2.6 software. Trichostatin A (TSA) was used as a reference to screen 19 small molecular components from TCM, and the default docking conformation number was set to obtain the docking binding energy, active site amino acid residues and hydrogen bonds, and the biological activity was verified. Result::The binding energies of 19 small molecule components from TCM to HDAC3 and HDAC8 were different. Among them, ursolic acid, fangchinoline and tetrandrine have low binding energies to HDAC3 and HDAC8, and their binding activities were strong. The optimal binding energy of fangchinoline and HDAC3 at the site 1 was the lowest (-26.71 kJ·mol-1), and that of HDAC8 at the site 9 was the lowest (-26.84 kJ·mol-1). The optimal binding energy of tetrandrine and HDAC3 at the site 13 was the lowest (-26.38 kJ·mol-1), and that of HDAC8 at the site 12 was the lowest (-25.41 kJ·mol-1). In addition, the binding energy of ursolic acid and HDAC3 at the site 16 was the lowest (-25.83 kJ·mol-1), and that of HDAC8 at the site 8 was the lowest (-35.62 kJ·mol-1). Three kinds of amino acids at the docking site of small molecules were rendered by PyMOL 2.3.1.When ursolic acid was combined with HDAC3/8, the active sites produced two hydrogen bonds, and the interaction was strong, and many amino acids were connected at the active site. The fangchinoline formed two hydrogen bonds with the active site of HDAC3 and one hydrogen bond with the active site of HDAC8, and hydrophobic binding with some active site amino acids. There was no hydrogen bond between tetrandrine and HDAC3/8, and all docking sites were docked by 4 active amino acids. Three small molecules (ursolic acid, fangchinoline and tetrandrine) with the best docking effect had the inhibitory activity against HDAC3/8 at the concentration of 500 μmol·L-1 and 100 μmol·L-1, and the inhibitory activity was still optimal among the 10 selected small molecules. Conclusion::Among the screened 19 small molecules, ursolic acid, tetrandrine and fangchinoline may be the new anti-inflammatory drugs of HDAC3/8 inhibitory target, which provides a reference for further exploration and discovery of new anti-inflammatory drugs.

UPLC fingerprint-based analysis of differences in chemical components from different parts of Stephania tetrandra / 中草药

Objective: To analyze and compare UPLC fingerprints of root, rhizome, stem, and leaf of Stephania tetrandra, learn the differences in chemical component types and contents of main active components, and provide basis for rational development and utilization of S. tetrandra. Methods: UPLC was used to obtain characteristic chromatograms of different parts; The Similarity Evaluation System for Chromatographic Fingerprints of Traditional Chinese Medicine (Version 2012) was run to capture the common peaks of different parts and calculate their similarity and analyze the characteristic peaks of different parts. SPSS 23.0 was run to compare the difference in component contents of the roots and rhizomes using the paired sample t-test. Results: The similarity in chemical composition between root and rhizome was 0.928, indicating they have similar chemical composition, and both of them contained tetrandrine and fangchinoline, the index components. The similarity between rhizome and leaf was 0.947; The similarity was low between stem, leaf and root and rhizome, and there were no tetrandrine and fangchinoline in the first two parts. The results of paired samples t-test show that the total content of chemical components in rhizome was higher than that in roots, and the mainly difference came from other non-index components, but there was no significant difference between tetrandrine and fangchinoline. Conclusion: Significant differences are present in chemical composition types and contents of different medicinal parts of S. tetrandra; The type of chemical components in rhizome is similar to that in root, and the content of some components in rhizome is significantly higher than that in root, which means that rhizomes can be used as an equivalent of roots. Stems and leaves cannot be a substitute for roots because they do not contain tetrandrine and fangchinoline, but they contain many other chemical components which can be utilized as a new resource.

Quality Regionalization and Environmental Impact Factors Analysis Based on Main Active Ingredients of Stephaniae Tetrandrae Radix / 中国实验方剂学杂志

Objective:To investigate the quality regionalization and environmental impact factors of Stephaniae Tetrandrae Radix based on main active ingredients，and provide a reference for the determination of high-quality production areas and the dominant environmental factors affecting the content of Stephaniae Tetrandrae Radix. Method:Partial least squares regression analysis (PLS) and principal component analysis (PCA) were used to study the quality regionalization and environmental impact factors based on the main active ingredients of tetrandrine and fangchinoline, and investigate the environmental factors of the producing areas. Result:The content of fangchinoline was positively correlated with soil pH and annual average temperature，negatively correlated with latitude. The content of tetrandrine was positively correlated with soil pH，negatively correlated with annual rainfall and longitude. The total content was positively correlated with soil pH and annual average temperature，but negatively correlated with annual rainfall，latitude and longitude. Principal component analysis showed that the 50 production areas could be divided into four groups of quality formation. The groups with the highest scores were Shixing county in Guangdong，Shexian county in Anhui，Songxi county in Fujian，Nanxiong city in Guangdong and Xiangxiang city in Hunan，all of which were best areas for accumulation of the two main active ingredients. Conclusion:Soil pH，annual average temperature，annual rainfall，latitude and longitude are the main environmental factors affecting the main active ingredients of Stephaniae Tetrandrae Radix. The best areas for accumulation of tetrandrine and fangchinoline are Shixing county in Guangdong，Shexian county in Anhui，Songxi county in Fujian，Nanxiong city in Guangdong and Xiangxiang city in Hunan.

Simultaneous Determination of Fangchinoline,Tetrandrine,Mesaconitine,Aconitine and Hypaconitine in Huoxue Zhentong Plaster by HPLC with Gradient Elution / 中国药师

Objective:To develop an HPLC gradient elution method for the simultaneous determination of fangchinoline,tetrandrine,mesaconitine,aconitine and hypaconitine in Huoxue Zhentong plaster.Methods::A Dikma-C18 (200 mm×4.6 mm,5 μm) chromatographic column was adopted,the mobile phase was methanol-acetonitrile (3∶1)(A)-0.06% diaethylamin solution (B) with gradient elution at a flow rate of 1.0 ml·min-1,the detection wavelength was 280 nm for fangchinoline and tetrandrine,and 235 nm for mesaconitine,aconitine and hypaconitine.The column temperature was set at 30 ℃,and the injection volume was 10 μl.Results::There was a good linear relationship when the content of fangchinoline,tetrandrine,mesaconitine,aconitine and hypaconitine was within the range of 7.490-149.800 μg·ml-1(r=0.999 9),14.610-292.200 μg·ml-1(r=0.999 8),4.150-83.000 μg·ml-1(r=0.999 2),5.250-105.000 μg·ml-1(r=0.999 6) and 5.140-102.800 μg·ml-1(r=0.999 9),respectively.The average recovery and the corresponding RSD were 99.87%(0.49%),97.79%(1.11%),96.97%(1.75%),98.60%(1.50%) and 97.94%(0.98%)(n=6),respectively.Conclusion:An HPLC gradient elution method is successfully established for the simultaneous determination of 5 components in Huoxue Zhentong plaster.The established method is simple,accurate and reliable,which is helpful to the quality control of Huoxue Zhentong plaster.

Simultaneous determination of darutoside, kirenol, darutigenol, fangchinoline and d-tetrandrine in xixian fengshi wan by joint wavelength switching HPLC gradient elution method / 第二军医大学学报

Objective To establish a joint wavelength switching HPLC gradient elution method for simultaneous determination of darutoside, kirenol, darutigenol, fangchinoline and d-tetrandrine in Xixian Fengshi Wan. Methods A Kromasil C18 column (4. 6 mm × 250 mm, 5 μm) was used with a mobile phase A; acetonitrile-methano (2: 1) and mobile phase B: 0. 05% acetic acid solution with gradient elution. The flow rate was 1. 0 mL/min; the injection volume was 20 μL; darutoside, kirenol and darutigenol were detected at 215 nm, and fangchinoline and d-tetrandrine were detected at 280 nm. Results In the given concentration range, the linearity ranges of darutoside, kirenol, darutigenol, fangchinoline and d-tetrandrine were 6. 45-129. 00 μg/mL (r=0. 999 5), 5. 61-112. 20 μg/mL (r=0. 999 9), 4. 25-85. 00 μg/mL (r=0. 999 3), 9. 19-183. 80 μg/mL (r =0.999 8), and 11. 05-221. 00 μg/mL (r=0. 999 7), respectively; and their average recoveries and RSD were 98. 73% (1. 43%), 97. 63% (1. 28%), 99. 44% (1. 29%), 98. 33% (1. 38%), and 97. 36% (1. 37%), respectively. Conclusion The established joint wavelength switching HPLC gradient elution method is simple, accurate and reproducible; it can simultaneously determine the contents of darutoside, kirenol, darutigenol, fangchinoline and d-tetrandrine in Xixian Fengshi Wnn and can be used for quality control of Xixinn Fengshi Wan.

Determination of tetrandrine and fangchinoline in Qingshenjianfei Tablets by RP-HPLC / 中成药

AIM: To determine the contents of tetrandrine and fangchinoline in Qingshenjianfei Tablets by RP-HPLC. METHODS:In this method Zorbax C 18 column was used, and methanol—0.3%diethylamine (75∶25) as a mobile phase , the detection wavelength was at 242 nm. RESULTS:The recovery of tetrandrine was 103.65% and RSD was 1.59% .The recovery of fangchinoline was 97.11% and RSD was 1.91% (n=6). CONCLUSION: This method is simple,quick,reproducible and can be used for the quantitative analysis。

Simultaneous determination of tetrandrine,fangchinoline and berberine hydrochloride in Fengtongan Capsule by RP-HPLC / 中成药

AIM:To develop a method for simultaneously determining tetrandrine,fangchinoline and berberine hydrochloride in Fengtongan Capsule(Radix Stephaniae tetrandrae,Medulla Tetrapanacis,Fructus Chaenomelis,Cortex Phellodendri Chinensis,etc).METHODS:RP-HPLC was used with Phenomenex Gemini C_ 18 column.The mobile phase was a mixture of methanol-acetonitrile-phosphate buffer solution(0.01 mol/L potassium dihydrogen phosphate and 0.01 mol/L sodium heptanesulfonate were mixed equally,which contained 0.2% triethylamine,then was adjusted to pH 3.0 by phosphoric acid)(25∶20∶55).The detection wavelength was at 230 nm.RESULTS:There was a good linear relationship in tetrandrine content of 0.051 52-0.824 3 ?g(r= 0.999 9),fangchinoline content of 0.035 20-0.563 2 ?g(r= 0.999 9),and berberine hydrochloride content of 0.031 36-0.501 8 ?g(r= 0.999 7).The average recovery of tetrandrine was 97.23%(RSD=1.3%,n=9),99.56%(RSD= 1.6%,n=9)for fangchinoline and 98.22%(RSD=1.5%,n=9)for berberine hydrochloride.CONCLUSION:The method is simple,selective and reproducible.It can be applied to the quality control of Fengtongan Capsule.