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Phytochemical investigation on the ethanol extract of a well-known medicinal herb Leonurus japonicus, led to the separation of 18 labdane type diterpenoids (1-18). Through comprehensive spectroscopic analyses and quantum chemical calculations, these compounds were structurally characterized as six new interesting 5,5,5-di-spirocyclic ones (1-6), two new (7 and 8) and six known (13-18) interesting 6,5,5-di-spirocyclic ones, a new rare 14,15-dinor derivative (9), and three new ones incorporating a γ-lactone unit (10-12). An in vitro neuroprotective assay in RSC96 cells revealed that compounds 7 and 12 exhibited neuroprotective activity in a concentration-dependent way, comparable to the reference drug N-acetylcysteine.
Subject(s)
Magnetic Resonance Spectroscopy , Leonurus/chemistry , Plants, Medicinal , Diterpenes/chemistry , Plant Components, Aerial , Molecular StructureABSTRACT
OBJECTIVE To establish HPLC finger print of Leonurus japonicus granules，and to determine the contents of 4 index components such as leonurine hydrochloride ，ferulic acid ，rutin，hyperoside. METHODS The determination was performed on Inertsil TM ODS-3 column with mobile phase consisted of acetonitrile （A）-0.1％ formic acid solution （B）in the form of gradient elution；the flow rate was 1.0 mL/min，the detection wavelength was 280 nm，the column temperature was 25 ℃，and the sample size was 5 µL. Similarity Evaluation System of Chromatogram Fingerprint of TCM （2012 edition）was used for establishing the HPLC fingerprints of 10 batches of L. japonicus granules and analyzing their similarities. By comparing with HPLC fingerprints of reference substance ，the common peaks were identified. SPSS 25.0 and SIMCA 13.0 software were used for cluster analysis and principal component analysis ；the above HPLC method was used for the content determination of 4 index components in L. japonicus granules such as leonurine hydrochloride ，ferulic acid ，rutin，hyperoside. RESULTS HPLC fingerprints of 10 batches of L. japonicus granules were established ，and 16 common peaks were matched ，and 4 peaks identified were leonurine hydrochloride （peak 6），ferulic acid （peak 13），rutin（peak 14），hyperoside（peak 16）；the similarities of 10 batches of samples were all higher than 0.970. The 10 batches of samples could be divided into four categories by cluster analysis and principal component analysis；the classification results were consistent. The contents of leonurine hydrochloride ，ferulic acid ，rutin and hyperoside were 122.10-138.82 μ g/g，9.33-10.45 μ g/g，14.12-18.95 μ g/g，5.87-8.06 μ g/g，respectively. CONCLUSIONS Established HPLC fingerprint of L. japonicus granules and the method for the content determination of 4 index components are simple and easy to operate，and have high precision and good repeatability ，which provide reference for the quality evaluation of L. japonicus granules.
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Objective: To study the content difference of multi-index components between Yao medicine Young Leonurus heterophyllus (YLH) and Leonurus japonicus, and evaluate the quality with the content. Methods: A high performance liquid chromatography (HPLC) method was established for the simultaneous content determination of five chemical constituents of chlorogenic acid, leonurine hydrochloride, rutin, hyperin and isoquercitrin in Yao medicine YLH and Leonurus japonicus. SPSS 25.0 statistical software was used to conduct paired samples t-test and one-way ANOVA to infer the content differences among the samples. Principal component analysis (PCA) was used to evaluate the quality of medicinal materials. Results: T-test results of paired samples of Yao medicine YLH and Leonurus japonicus showed that there was significant difference in rutin content among the five chemical constituents, with no significant difference in other constituents. The results of one-way ANOVA analysis showed that there were significant differences in five chemical constituents of Yao medicine YLH and Leonurus japonicus from different habitats (P < 0.01). The results of principal component analysis showed that the higher scores of Leonurus japonicus from different habitats were from Nanning and Yulin, with the lowest in Shangsi County. Conclusion: There is a significant difference between the content of Yao medicine YLH and Leonurus japonicus. The quality of medicinal materials from Nanning and Yulin is better. The results provide data support for the comparison of the content between two kinds of Leonurus heterophyllus and Leonurus japonicus from different habitats.
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Objective:Computer network pharmacology technology was used to screen the main active ingredients of Tripterygium hypoglaucum radix-Leonurus japonicus herba for the treatment of rheumatoid arthritis(RA), predict the targets of the active ingredients, establish a pharmaceutical ingredient-active ingredient-target network, and further explore the potential mechanism of Tripterygium hypoglaucum radix-Leonurus japonicus herba for the treatment of RA. Method:RA disease targets were collected through DisGeNET, TTD, and Drugbank databases, the potential active components of Tripterygium hypoglaucum radix and Leonurus japonicus herba and their corresponding targets were obtained from the Chinese Medicine System Pharmacology Analysis Platform (TCMSP); common targets for drugs and diseases were screened by using the ImageGP platform; a common target interaction (PPI) network model was constructed by using the String database, a "drug-active ingredient-key target" network was constructed by using Cytoscape software, a protein interaction network was constructed by using the String database, gene function (GO) analysis and pathway enrichment analysis based on the Kyoto Gene and Genomic Encyclopedia (KEGG) were performed by using the ClueGO plug-in. Result:Through screening, 9 active pharmaceutical ingredients were obtained, involving a total of 235 targets, and 7 active ingredients were related to the disease targets. 24 common targets for Tripterygium hypoglaucum radix Leonurus japonicus herba-disease were obtained. The common targets were mainly enriched in 278 biological processes and 141 signaling pathways to play a role in the treatment of RA. Conclusion:The therapeutic effect of Tripterygium hypoglaucum radix Leonurus japonicus herba on RA reflects the characteristics of multi-component-multi-target-multi-channel of traditional Chinese medicine, and provides a scientific basis for explaining its mechanism and clinical application of RA.
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An HPLC fingerprint and multi-component determination method of Leonurus japonicus was established for comprehensive evaluation and quality control of Leonurus japonicus. The sample was incubated in 70% ethanol in a water bath for 2 h, and the extract was analyzed by HPLC using Kromasil C18 column (250 mm×4.6 mm, 5 μm). The mobile phase consisted of acetonitrile-0.1% formic acid with gradient elution. The flow rate was 1.0 mL·min-1. The temperature of column was 30 ℃. The detection wavelength was 280 nm. HPLC fingerprint of characteristic components of Leonurus japonicus was established. There were 12 common peaks among 25 batches of samples, and 5 of them were identified and determined. Syringic acid, leonurine hydrochloride, rutin, hyperoside or isoquercitrin showed a good linearity in the ranges of 0.426 1-85.22 ng (r = 0.999 9), 7.948-1 590 ng (r = 0.999 3), 10.20-2 040 ng (r = 1.000 0), 2.018-403.6 ng (r = 0.999 9), or 8.704-1 741 ng (r = 0.999 9), respectively. The average recoveries were 99.0%, 97.6%, 97.4%, 96.9% and 98.5% with RSD of 1.1%, 1.8%, 1.4%, 1.5% and 1.3%, respectively. The HPLC characteristic fingerprint of Leonurus japonicus was specific, and this method can simultaneously determine the content of 5 components.
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Objective: Crude Leonuri Fructus (CLF), the fruits of the Leonurus japonicus Houtt, and processed Leonuri Fructus (PLF) by stir-baking as the important Chinese herbal medicines, have been used in China and other Asian countries for thousands of years. The objective of this research is to reveal the difference between CLF and PLF. Methods: The sensory technologies of the colorimetry, sensitive and validated HPLC-ELSD and GC combined with flame ionization detector (GC-FID) were employed to discriminate CLF and its processed product PLF. The color parameters of the samples were determined by colorimetric instrument CR-410. Moreover, the content of stachydrine and six fatty acids were determined by HPLC and GC. Subsequently, analysis of variance (ANOVA), principal components analysis (PCA), hierarchical cluster analysis (HCA), and Kendall's correlation test were performed for data analysis. Results: The CLF and PLF were divided into two categories by PCA and HCA in terms of their component content and color. The results distinctly demonstrated significant changes in color and the content of indicative components between CLF and PLF. Conclusion: The study revealed that HPLC, GC, and colorimetric method in combination with chemometric method could be used as comprehensive quality evaluation for CLF and PLF.
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Objective: To establish a simple and effective method for the isolation of leonurine from Leonurus japonicus by high speed countercurrent chromatography (HSCCC). Methods: The extraction conditions of leonurine were optimized by one-factor experimental design. After comparing several different solvent systems, the two phase system of ethyl acetate-n-butanol-water (3∶2∶5) was finally chosen as operating solvent of HSCCC for the separation of leonurine, in which the lower phase was determined as the mobile phase and the upper phase as stationary. The detection of eluates was performed with an ultraviolet detector at 277 nm. The rotation speed was adjusted at 850 r/min, and the flow rate was 2.2 mL/min. Results: Leonurine was successively isolated from n-butanol fraction by HSCCC, and the above established method was also successively applied to the crude extact of L. japonicus. Finally, 68 mg of leonurine with purity about 96.2% could be obtained from 2.48 g crude extract of L. japonicus in a single injection. Conclusion: The described approaches actively promote efficient preparation strategy to obtain high purity of leonurine.
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Objective To find the genuine structure with anti-acetylcholinesterase (anti-AChE) from the phenolic glycosides abundant in Leonurus japonicus (Lamiaceae). The assay for anti-AChE activity is often used to lead anti-Alzheimer's drugs. Methods The five phenolic glycosides, tiliroside, leonurusoside C, 2‴-syringoylrutin, rutin, and lavanduliofolioside were isolated from L. japonicus. The activities of the glycosides were relatively low. Seven compounds including p-coumaric acid, caffeic acid, hydroxytyrosol, salidroside, syringic acid, kaempferol, and quercetin, which are produced by the hydrolysis of the five glycosides, were also assayed for anti-AChE activity. Results Of those seven compounds, the five compounds other than salidroside and syringic acid exhibited potent anti-AChE activities. In particular, the IC
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Three new labdane diterpenoids, leojaponicone A (1), isoleojaponicone A (2) and methylisoleojaponicone A (3), were isolated from the herb of Leonurus japonicus. The chemical structures of these secondary metabolites were elucidated on the basis of 1D and 2D NMR, including HMQC, and HMBC spectroscopic techniques. All the new compounds were tested in vitro for their acetylcholinesterase and α-glucosidase inhibitory activity. Compounds 1-3 exhibited low inhibitory effects on α-glucosidase with respect to acarbose and exhibited high inhibitory effects on acetylcholinesterase with respect to huperzine A.
Subject(s)
Acetylcholinesterase , Metabolism , Cholinesterase Inhibitors , Chemistry , Pharmacology , Diterpenes , Chemistry , Pharmacology , Glycoside Hydrolase Inhibitors , Chemistry , Pharmacology , Leonurus , Chemistry , Magnetic Resonance Spectroscopy , Molecular Structure , Plant Extracts , Chemistry , PharmacologyABSTRACT
Objective: To find the genuine structure with anti-acetylcholinesterase(anti-AChE)from the phenolic glycosides abundant in Leonurus japonicus(Lamiaceae).The assay for anti-AChE activity is often used to lead anti-Alzheimer's drugs. Methods: The five phenolic glycosides, tiliroside, leonurusoside C, 2'''-syringoylrutin, rutin, and lavanduliofolioside were isolated from L. japonicus. The activities of the glycosides were relatively low. Seven compounds including p-coumaric acid, caffeic acid, hydroxytyrosol, salidroside, syringic acid, kaempferol, and quercetin, which are produced by the hydrolysis of the five glycosides, were also assayed for anti-AChE activity. Results: Of those seven compounds, the five compounds other than salidroside and syringic acid exhibited potent anti-AChE activities.In particular,the IC50s of caffeic acid and quercetin were (1.05 ± 0.19)and (3.58 ± 0.02)μg/mL, respectively. Rutin was the most abundant flavonoid in the extract(9.18 mg/g as measured by HPLC). Conclusion: The substances with potent anti-AChE were caffeic acid, quercetin, p-coumaric acid, kaempferol, and hydroxytyrosol that can be produced from their glycosides.
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Quality control of Chinese materia medica (CMM), a key factor in restricting the development of modernization of CMM, has always been a hot issue to traditional Chinese medicine (TCM) circles, our society, and the public. At present, determination of chemical components is the main means for quality control of vast majority of CMM, but many components lack not only specificity, but also biological activities. This is bound to greatly reduce the value of quality standard of CMM, which makes it difficult to reflect the true quality of CMM. Quality marker (Q-Marker) of CMM is a new concept that brings forward higher requirement to quality control of CMM. Based on the basic conditions of Q-Marker, Leonurus japonicus and Penthorum chinense have been investigated systematically. This paper introduces theIR research processes from five aspects: effective material basis, specificity of the isolated components, chemical structure and bioactivity, measurability, and fingerprint spectrum.
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The present study aimed at isolation and purification of the bioactive terpenoids from the herb of Leonurus japonicus by chromatographic separations such as silica gel, sephadex LH-20 and C18 reversed phase silica gel, as well as preparative HPLC. As a result, leojaponic acids A (1, C17H24O4) and B (2, C18H26O4), two homologous terpenoids, together with (-)-loliolide (3), 1-(3-ethylphenyl) ethane-1, 2-diol (4) and dibutyl phthalate (5), were isolated from the EtOH extract of L. japonicus. All the chemical structures of the isolates were elucidated on the basis of 1D and 2D NMR analyses. Compounds 1 and 2 were new terpenoids, and Compounds 3 and 4 were isolated and identified for the first time from this plant. In addition, the α-glucosidase and tyrosinase inhibitory activity of the new compounds were evaluated.
Subject(s)
Enzyme Inhibitors , Chemistry , Fruit , Chemistry , Glucosidases , Leonurus , Chemistry , Magnetic Resonance Spectroscopy , Molecular Structure , Plant Extracts , Chemistry , Terpenes , ChemistryABSTRACT
OBJECTIVE:To observe therapeutic efficacy of Compound Leonurus japonicus oral solution in the treatment of hemorrhage after artificial abortion and its effect and safety on normal menstruation recovery. METHODS:100 cases of artificial abortion selected and randomly divided into treatment group and control group,with 50 cases in each group. Control group re-ceived oxytocin intramuscularly after surgery,10 U/time,12 h/time,for 3 times;treatment group was additionally given Com-pound L. japonicus oral solution,2 piece/time,3 times/d,for 5 days,on the basis of control group. Metrorrhagia duration,hemor-rhage amount,normal menstruation recovery and duration of abdominal pain and ADR were all observed in 2 groups. RESULTS:The metrorrhagia duration [(5.38±0.95)d] of treatment group was significantly lower than that [(8.58±1.32)d] of control group, with statistical significance(P<0.05);hypomenorrhea cases(24 cases)of treatment group was higher than those(9 cases)of con-trol group;menorrhagia cases(8 cases)was lower than those(24 cases)of control group;with statistical significance(P<0.05). The normal menstruation recovery time [(27.34±3.87)d] of treatment group was significantly shorter than that [(40.13±2.58)d] of control group,with statistical significance(P<0.05);the duration of abdominal pain [(2.85±0.89)d] in treatment group was sig-nificantly shorter than [(5.02 ± 1.33)d] in control group,with statistical significance (P<0.05).Obvious ADR was not found in both groups. CONCLUSIONS:Compound L. japonicus oral solution is effective in the treatment of hemorrhage after artificial abor-tion,can significantl shorten metrorrhagia duration and hemorrhage amount,recover normal menstruation early,improve the symp-toms of abdominal pain after operation and is easy to take and good safety.
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Objective: To study the low polar chemical constituents from Leonurus japonicus. Methods: The chemical constituents were isolated and purified by various chromatographic methods including column chromatography over silica, Sephadex LH-20, and Rp C18 columns. Their structures were identified by physicochemical properties and spectroscopic analysis. Results: Six aliphatic compounds were isolated from the EtOAc extracts of L. japonicus, including two allenes: (-)-nonadeca-5,6-dienoic acid methyl ester (1) and methyl octadeca-5,6-dienoate (2); four saturated aliphatic compounds: heneicosanoic acid (3), arachidic acid (4), heptacosanoic acid (5), and methyl myristate (6). Conclusion: All the compounds are isolated from L. japonicus for the first time. Compound 1 is a new one named leonuallenote A, while compound 2 is first reported from the plants of Leonurus L.
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Objective: To study the chemical constituents from Leonurus japonicus Injection. Methods: The constituents were purified by macroporus resin, silica gel, Sephadex LH-20, and recrystallization. The structures were identified by spectroscopic methods and spectroscopic analysis. Results: Fifteen compounds were isolated from L. japonicus Injection, including four amino acids and their derivatives such as N-isobutyl valine (1), valine (2), alanine (3), L-pyroglutamic acid methyl ester (4), nine alkaloids: stachydrine (5), choline (6), trigonelline (7), uracil (8), 5-methyluracil (9), 2', 3'-O-isopropylidene uridine (10), 3-hydroxypyridine (11), 5-hydroxyl-2-hydroxymethylpyridine (12), and 2-methyl-3-hydroxypyridine (13); and two furoic acids such as furan-2- carboxylic acid (14) and 5-hydroxymethyl-2-furancarboxyic acid (15). Conclusion: In addition to compounds 5 and 8, the others are isolated from the plants of Leonurus Linn. for the first time. Among them, compound 1 is a natural product. Its NMR data, undoubtedly assigned by 2D-NMR, have been reported for the first time.
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Objective To establish the quality standard for Chanhou Zhuyu Capsules.Methods Leonurus Japonicus and Ligusticum chuanxiong in Chanhou Zhuyu Capsules were identified by TLC,and the content of stachydrine hydrochloride was determined by TLC Scanning.Results The quality identification by TLC was specific.Stachydrine hydrochloride had a good linearity in the range of 4.0～12.0 ?g,r=0.999 7,and the average recovery was 99.87 %,RSD=1.67 %.Conclusion The methods of identification and content determination are simple,reliable and reproducible.They can be used effectively for the quality control of Chanhou Zhuyu Capsules.