Minor triterpenoid acids from an aqueous extract of Uncaria rhynchophylla / 药学学报

Seventeen minor triterpenoid acids (1-17) were isolated from an aqueous decoction of Uncaria rhynchophylla by a combinatory application of column chromatography using multiple stationary phases, including macroporous adsorbent resin, MCI resin, Sephadex LH-20, Toyopearl HW-40C, silica gel, and C18 reversed phase silica gel, combined with separation techniques of flash chromatography (FC) and high performance liquid chromatography (HPLC). Their structures were determined by analysis of HR-ESI-MS, UV, CD, and IR as well as 1D and 2D NMR spectroscopic data, of which eight new compounds (1-8) are named successively uncarinic acids Q-X, while the structures of 2 and 7 were confirmed by single crystal X-ray diffraction. In the in vitro assays, 27-hydroxyolean-12-en-28-oic acid (17) inhibited TGF-β-induced HSC-T6 cell activation at the concentration of 5 μmol·L-1.

Megastigmanes from an aqueous extract of Uncaria rhynchophylla / 药学学报

Five new megastigmanes (1-5) were isolated from a decoction of Uncaria rhynchophylla by separation techniques of column chromatography using a combination of multiple stationary phases, including macroporous adsorbent resin, MCI resin, silica gel, Sephadex LH-20, and Toyopearl HW-40F, and reversed phase HPLC. Their structures were characterized by spectroscopic data analysis of HR-ESI-MS, NMR, and CD, in combination with Mosher's mothed as well as ECD and NMR calculations. The new compounds were named uncarphyllonone A (1), uncarphyllonols A (2) and B (3), and uncarphabscisic acids A (4) and B (5). Although the structures of 3 and 4 were previously reported, the reported NMR spectroscopic data were incorrect or do not support the assigned structures in literatures. This is also the first report of discovery of new megastigmane natural products from the Uncaria genus.

Steroids from Uncaria rhynchophylla / 中国中药杂志

Thirteen steroids(1-13) were isolated from the non-alkaloid constituents of Uncaria rhynchophylla by column chromatography on silica gel, ODS, Sephadex LH-20, and preparative HPLC chromatography, and their structures were elucidated by analyses of the MS and NMR spectral data. All the compounds were isolated from the genus Uncaria for the first time, and 1 was a new compound. The ~1H-NMR and ~(13)C-NMR data of two compounds(12 and 13) in deuteron-chloroform were completely assigned. This study enriched the steroid constituents of U. rhynchophylla and provided scientific references for the elucidation of active constituents and further development and utilization of U. rhynchophylla.

Cloning and analysis of STR gene and its promoter from Uncaria / 药学学报

On the basis of the Uncaria transcriptome, specific primers were designed for UrSTR. The full-length cDNA of UrSTR (GeneBank: OL310251) was 1 541 bp, encoding 345 amino acid residues, and the promoter region sequence of UrSTR (GeneBank: OL310252) was 1 179 bp. Phylogenetic tree is revealed that UrSTR had a closest relationship with STR from Ophiorrhiza pumila and Ophiorrhiza japonica. Localization of UrSTR protein is revealed located in the vacuole membrane. Plant-care analysis indicated that the promoter region sequence of UrSTR, covering multiple light, stress and hormone-response cis-regulatory elements, and verified transcriptional activity. The results of SDS-PAGE show that pET-28a-UrSTR recombinant protein was successfully expressed, and the size was anticipated. The UrSTR prokaryotic expression system needs to be optimized in the later stage. The research lays the foundation for further purification to study its structure and functional characterization of the UrSTR protein.

Evaluation of the effects of Uncaria rhynchophylla alkaloid extract on LPS-induced preeclampsia symptoms and inflammation in a pregnant rat model

Excessive pro-inflammatory cytokines result in adverse pregnancy outcomes, including preeclampsia-like phenotypes, and fetal growth restriction. Anti-inflammation might be an effective therapy. The aim of this research was to investigate whether Uncaria rhynchophylla alkaloid extract (URE), a highly safe anti-inflammation constituent of the herb, can inhibit inflammation and improve clinical characteristics of preeclampsia in a lipopolysaccharide (LPS)-induced preeclampsia rat model. The rat model was established by daily administration of LPS (1 μg/kg body weight per day) from gestational day (GD) 14 to 19. Different doses of URE (35, 70, and 140 mg/kg body weight per day) were administered from GD 14 to GD 19. The effects of URE on proteinuria, maternal hypertension, pregnancy outcomes, as well as pro-inflammatory cytokines levels in serum and placenta were measured. High-dose URE (HURE) treatment decreased LPS-induced mean 24-h proteinuria and systolic blood pressure, and increased fetal weight, placental weight, and the number of live pups (P<0.05). Moreover, increased serum and placental levels of interleukin (IL)-6, IL-1β, tumor necrosis factor-α, and interferon-γ in the LPS-treated group were obviously inhibited after HURE administration (P<0.01). URE improved preeclampsia symptoms and mitigated inflammatory responses in the LPS-induced preeclampsia rat model, which suggests that the anti-inflammation effect of URE might be an alternative therapy for preeclampsia.

Two folate-derived analogues from an aqueous decoction of Uncaria rhynchophylla / 中国天然药物

Two new folate-derived analogues, named uncarophyllofolic acids A (1) and B (2), respectively, were isolated from the Uncaria rhynchophylla hook bearing stem (Gouteng in Chinese). The distinct stereochemical structures of 1 and 2 were determined by spectroscopic data analysis in combination with acidic hydrolysis and Marfey's derivatization, along with comparison of their specific rotation and Cotton effect (CE) data with those of the biogenetically related known derivatives as well as theoretical calculations of electronic circular dichroism (ECD) spectra. A plausible biosynthetic pathway of 1 and 2, associating to folate metabolism and the previously reported orychophragines A-C from Orychophragmus violaceus, is discussed.

Alkaloids from an aqueous extract of Uncaria rhynchophylla / 药学学报

Five alkaloids were isolated from a decoction of Uncaria rhynchophylla by a combination of various chromatographic techniques, including macroporous adsorbent resin, MCI resin, silica gel, Sephadex LH-20, and reversed phase HPLC. Their structures were characterized by comprehensive analyses of spectroscopic data as monoterpene indole alkaloids (+)-(7R)-3-oxo-7-hydroxy-3,7-seco-dihydrorhynchohylline (1), (+)-(7S)-3-oxo-7-hydroxy-3,7-seco-dihydrorhyncho-hylline (2), (+)-(7R)-3-oxo-7-hydroxy-3,7-seco-rhynchohylline (3) and (+)-(7S)-3-oxo-7-hydroxy-3,7-seco-rhynchohylline (4), and a β-carboline alkaloid 1,2,3,4-tetrahydro-1-oxo-β-carboline (5). Among them, 1 and 2 are new compounds, 3 and 4 are new natural products that were semi-synthesized from isorhynchohylline with incorrect specific rotations, and 5 is isolated for the first time from the genus Uncaria.

Research progresses in pharmacological effects of rhynchophylline and its isomers on nervous system / 药学学报

Uncaria rhynchophylla is one of the frequently used herbs in China, it is mainly used for heat-clearance, suppression of hyperactive liver, calming endogenous wind and arresting convulsion in traditional Chinese medicine (TCM). Alkaloids are the main active materials in Uncaria rhynchophylla, pharmacological studies have shown that Uncaria rhynchophylla and its alkaloids have comprehensive biological effects on the nervous system. Rhynchophylline is one of the most abundant alkaloids in Uncaria rhynchophylla. The recent studies demonstrate that rhynchophylline and its isomers (isorhynchophylline, corynoxine, corynoxine B) may be good drug candidates for treatment of Alzheimer's disease, Parkinson's disease, epilepsy, etc. Although the structures of the 4 alkaloids are very similar, they have different effects on nervous system. For example, corynoxine and corynoxine B exhibit better sedative effects than isorhynchophylline. Rhynchophylline and isorhynchophylline have been extensively studied. For development and utilization of rhynchophylline for nervous system disease, more studies are needed to unveil the structure-function relationship and the underlying mechanisms. Here, we summarizes the progresses the effects of rhynchophylline and its isomers on the nervous system.

Chemical constituents from leaves of Uncaria rhynchophylla / 中草药

Objective: To study the chemical constituents from the leaves of Uncaria rhynchophylla and their antioxidant activities. Methods: The compounds were isolated by various column chromatography. Their structures were identified by spectroscopic methods including 1H-NMR, 13C-NMR, MS, HR-MS, etc. The antioxidant activities were evaluated by DPPH radical scavenging assay. Results: Seventeen compounds were isolated from the leaves of U. rhynchophylla, and identified as 18, 19-hydroxy- vincosamide (1), α-tocopherol (2), α-tocospiro A (3), dihydroactinidiolide (4), ursolic acid lactone (5), 3, 4-dehydrotheaspirone (6), chakyunglupulin A (7), ursolic acid (8), protocatechuic acid (9), quercetin (10), epicatechin (11), chlorogenic acid ethyl ester (12), quercetin-3-O-robinobioside (13), rutin (14), methyl caffeate (15), strictosamide (16), and vincosamide (17). The anti-oxidative IC50 values of compounds 10, 11, and 13-15 were 7.52, 8.21, 8.14, 2.13, and 5.35 μmol/L, respectively, which were better than the positive control Vit E (IC50 value was 9.53 μmol/L). Conclusion: Compound 1 is a new compound named as vincosamide A, and compounds 2-7, 9, 12, 13, and 15 are isolated from genus Uncaria Schreber nom. cons. for the first time. Moreover, compounds 10, 11, and 13-15 have the certain prospects in the development of natural anti-oxidative agents.

Profiling on the Invention Patents of Uncaria rhynchophylla in China / 世界科学技术-中医药现代化

U.rhynchophylla is an important herb in Chinese medical treasure with the functions of calming endogenous wind and arresting convulsion,clearing away heat and calming the liver,for the treatments of hypertension,epilepsy and convulsion,crying with fear in children.This paper retrieved and collated 3,304 items of application information in the invention patents over uncaria before July,2016.Data of the basic sciences were collected from Chinese Science Citation Database (CSCD) and those of situation analysis of patents from Patent Online Analysis System of Chinese Academy of Sciences.Year,region,patented technology,patentee and key technology were involved and analyzed in the system for profound investigation of the current situation and developing trend of invention patent application over uncaria.It was found that trace elements and chemical composition beneficial to human health were receiving favors from the market with rapt attention focusing on patent applications.However,there were still major limitations in the application and the patent quality needed improving.In conclusion,further development and the utilization of uncaria presented promising prospects.

Evaluation and cumulative characteristics of heavy metals in soil-Uncaria rhynchophylla system of different functional areas / 中国中药杂志

Soil and Uncaria rhynchophylla in different functional areas were selected for the study,the content of heavy metals such as As, Cd, Cu, Cr, Pb, and Hg in soil and U. rhynchophylla was discussed, the characteristics of their accumulation in the U.rhynchophylla was analyzed, the contamination levels of heavy metals in soil in different functional areas was evaluated. The results showed that content of Cu, As, Pb and Cr in soil was being cropland>woodland>wasteland, content of Cd was being woodland>cropland>wasteland, content of Hg was being cropland>woodland>wasteland. According to quality standard of soil environment, soil Cd in woodland, cropland and wasteland all exceeded the state-level standards, soil Cd in woodland exceeded the secondary standard, soil Hg in cropland and wasteland all exceeded the state-level standards. According to technical conditions of green food producing area, soil Cd in woodland exceeded the limit value of standard. According to Green Trade Standards of Importing Exporting Medicinal Plants Preparations,the content of heavy metals of U.rhynchophylla in cropland,woodland and wasteland were correspond to the specification. From the single factor pollution index, the soil in woodland was polluted by Cd. From the comprehensive pollution index, the soils in different functional areas were not contaminated by heavy metals. The enrichment coefficient of heavy metals such as As, Cu, Cr, and Pb in hook of U.rhynchophylla was being wasteland>woodland>cropland, the enrichment coefficient of Cu in hook of U. rhynchophylla in wasteland was more than 1. Except Cu, the enrichment coefficient of other heavy metals was low.

Two new ortho benzoquinones from Uncaria rhynchophylla / 中国天然药物

The present study was designed to determine the chemical constituents of the stems and hooks of Uncaria rhynchophylla. The chemical constituents were isolated and purified from CH2Cl2 fraction by chromatography. Their structures were elucidated by spectroscopic analyses. Their cytotoxicity was tested using MTT method. Two new ortho benzoquinones, 3-diethylamino-5-methoxy-1, 2-benzoquinone (1) and 3-ethylamino-5-methoxy-1, 2-benzoquinone (2), together with a known compound isorhynchophyllic acid (3) were isolated from U. rhynchophylla. These compounds were evaluated for their cytotoxicity against cancer cells A549, HepG2 and A2780. Compounds 1 and 2 were new ortho benzoquinones and showed weak antiproliferative activities on A549, HepG2 and A2780 cells. Compound 3 significantly inhibited the proliferation of A549, HepG2 and A2780 cells with IC50 values being 5.8, 12.8 and 11.8 µmol·L(-1), respectively.

Analysis of the isorhynchophylline in Uncaria rhynchophylla from different regions and different harvest time / 国际中医中药杂志

Objective To study the content of isorhynchophylline from different regions and different harvest time.Methods The content of isorhynchophylline in Uncaria rhynchophylla from different regions and different harvest time was tested by HPLC. The Waters Symmetry C18 color (4.6 mm× 250 mm, 5 μm), mobile phase of methanol - 0.01 mol/L ammonium acetate buffer (pH 8.0) (60:40), column temperature 25℃, 20 μl sample volume, velocity of 1.0 ml/min, detection wavelength of 246 nm were setted.Results The isorhynchophylline can be detected in different regions and different harvest time, with the highest content was found in Ningming county and from September to February of the next year.Conclusion The content of isorhynchophylline in Uncaria rhynchophylla varied from different regions and harvest times. The best harvest time is during September to February of the next year.

Ameliorating effect of new constituents from the hooks of Uncaria rhynchophylla on scopolamine-induced memory impairment / 中国天然药物

AIM@#To study the chemical constituents and their anti-amnesic effect from the hooks of Uncaria rhynchophylla.@*METHODS@#The isolation of compounds was performed by chromatographic techniques and their structures were identified on the basis of spectral analysis. Their ameliorating effects on scopolamine-induced memory impairment in vivo using a Morris water-maze task and passive avoidance task system were evaluated.@*RESULTS@#Activity-guided fractionation of the total extracts resulted in the isolation of four constituents, trans-anethole (1), p-anisaldehyde (2), estragole (3), and 3-oxo-olean-12-en-28-oic acid (4), which were found for the first time from this plant.@*CONCLUSION@#Compound 1 exhibited a better memory enhancing effect than tacrine, a positive agent, at the same dose in the passive avoidance test and a similar property in the water-maze test, and its action may be mediated, in part, by the acetylcholine enhancing cholinergic nervous system.

Historical Studies about Medicinal Part of Chinese Crude Drug \lq\lqUncaria Hook\rq\rq (Part2) / 日本東洋医学雑誌

The crude Chinese drug “Uncaria hook” is a hook, or a twig with an attached hook of Uncaria plants seen in today's Chinese and Japanese medicinal markets. However, through herbological studies we found that the botanical origin of Uncaria hook was Uncaria rhynchophylla (Miq.) Miq., and that until the middle of Ming Dynasty in ancient China, the medicinal part used was the twig bark, not the hook itself, and use of the twig with hook was begun in the later Ming Dynasty. This change in practice seems to have been influenced by herbal descriptions written in the Ming Dynasty. Some of these stated that the medical effect of hooks was stronger than that of the bark.To determine the appropriate medicinal part of this crude drug in terms of chemical quality, we analyzed the alkaloid contents of Uncaria rhynchophylla bark, hooks, and twigs collected in Japan. Our result showed that the alkaloid content of the bark was higher than that of the twigs and hooks. Rhynchophylline and hirsutine, the alkaloid contained in Uncaria hook, were reported to improve memory learning and to cure hypertension, respectively. Since the alkaloid content profile of the bark was different from that of the hook, a question arose as to whether the medicinal properties of the part commonly used as “Uncaria hook” meet the requirement of the crude drug. Further pharmacological study is expected.

Historical Studies about Medicinal Part of Chinese Crude Drug \lq\lqUncaria Hook\rq\rq / 日本東洋医学雑誌

Japanese pharmacopoeia prescribes a crude drug, Uncaria hook, or the hook-like structures of Uncaria rhynchophylla (Miq.) Jacks, U. sinensis (Oliv.) Havil., and U. macrophylla Wall., while Chinese pharmacopoeia prescribes the hooks with attached stems of above3species, in addition to2other species of Uncaria hook; U. hirsuta Havil. and U. sessilifructus Roxb (all in the Rubiaceae family). Our herbological study has found that the botanical origin was Uncaria rhynchophylla before the Ming Dynasty, and that the part used was not the hook itself, but the plant's bark till the early Ming Dynasty. Use of the hooks with stems began in the late Ming Dynasty. On the other hand, in Japan, the hooks themselves have been mainly collected from wild U. rhynchophylla plants growing in southern warm-temperate zone of Japan, for use in Kampo medicines. We considered this differing Japanese custom was influenced by the descriptions in the Ben Cao Gan Mu (1596)written in the Ming dynasty by Li Si Chen, which state that the hook itself had medicinally sharp power. And we conclude that use of Uncaria bark alone is reasonable for prescriptions which originated before the early Ming dynasty, such as chotosan.

Different Hypotentive Effects of Various Active Constituents Isolated from Uncaria rhynchophylla / 中草药

Hypotensive effects of rhynchophylline, isorhynchophylline, total alkaloid and non-alka-loid fraction isolated from Uncaria rhynchophylla (Miq.) Jacks. were compared. Change in peripheralblood pressure was recorded by inserting a catheter into the right common carotid artery of anaethetizedrat, while each of the four hypotensive constituents was injected individually into the femoral vein by a mi-croinfusion pump. Results of the findings showed that the four constituents in U. rhynchophylla displayeddifferent hypotensive potency in the order of isorhynchophylline [lowering of mean arterial pressure(MAP) by 42.0%] > rhynchophylline (lowering of MAP by 32.1%) > total alkaloid (lowering of MAPby 21.3%) > non-alkaloid fraction (lowering of MAP by 12.4%). It was concluded that isorhyn-chophylline and rhynchophylline were the main hypotensive constituents in U. rhynchophylla.

Determination of tetrahydropalmatine and rhynchophylline and isorhynchophylline in Anshen Yangxue Oral Liquid by HPLC / 中成药

AIM: To establish the method for determining tetrahydropalmatine、rhynchophylline and isorhynchophylline in AnshenYangxue Oral Liquid(Ramulus uncariae cum uncis, stephania kwangsiansis,Radix polygoni multiflori praeparata cum succo glycinus sotae,Caulis Polygoni multiflori and Pine needle). METHODS: Tetrahydropalmatine、rhynchophylline and isorhynchophylline were determined by HPLC.Chromatographic condition was composed of Kromasil C_18 column,a mixture of methanol and water(55∶45) as mobile phase with 0.01 mol/L triehthylamine,adjusted with acetic acid to pH of 7.5,UV detection wavelength of rhynchophylline and isorhynchophylline was set at 254 nm,UV detection wavelength of tetrahydropalmatine was set at 281 nm. RESULTS: The averagere recoveries of tetrahydropalmatine、rhynchophylline and isorhynchophylline were 98.47%、99.04%and 98.75% respectively;RSD were 0.95%、2.6%and 1.6%,respectively. CONCLUSION: This method is simple,sensitive,accurate,and can be used for determining tetrahydropalmatine、rhynchophylline and isorhynchophylline in Anshen Yangxue Oral Liquid.