Isoverticine

Investigation of association of chemical profiles with the tracheobronchial relaxant activity of Chinese medicinal herb Beimu derived from various Fritillaria species.[Pubmed:28842340]

J Ethnopharmacol. 2018 Jan 10;210:39-46.

ETHNOPHARMACOLOGICAL RELEVANCE: Fritillariae Bulbus (Beimu in Chinese) is derived from the bulbus of many Fritillaria species (family Liliaceae), which has been used as an antitussive herb in traditional Chinese medicine for more than 2000 years. Due to the complexity of plant origins and significant variations in chemical profiles, the characterization of the profile of the major bioactive constituents and its association with pharmacological activity are important for the quality control of Beimu herbs from different origins. AIM OF THE STUDY: This study aims to investigate the distribution of major bioactive isosteroidal alkaloids in Beimu herbs of different origins and its correlation with the tracheobronchial relaxant activity. METHODS: Quantification of 7 main bioactive 5alpha-cevanine isosteroidal alkaloids, including ebeiedine, ebeiedinone, hupehenine, Isoverticine, verticine, verticinone and imperialine, in 23 Fritillaria species was performed using gas chromatography. The relaxant effect of different extracts of 4 commonly used Beimu herbs, namely Zhe-Beimu (F. thunbergii Miq.), Chuan-Beimu (F. cirrhosa D. Don), Hubei-Beimu (F. hupehensis Hsiao et K. C. Hsia) and Yi-Beimu (F. pallidiflora Schrenk), was evaluated using rat isolated tracheal and bronchial preparations pre-contracted with carbachol, the well established in vitro antitussive model. RESULTS: Amongst 23 Fritillaria species detected, significant variations of the types and quantities of 7 major isosteroidal alkaloids were determined, which served as an important indicator for the classification of different Beimu herbs with distinct geographic distributions. Based on the type and quantity of these alkaloids, different origins of Beimu could be clearly clustered into several subgroups by principal component analysis. Furthermore, both crude alkaloid and water extracts of all 4 Beimu herbs showed a dose-dependent tracheobronchial relaxation with different potencies. The total content of alkaloids (weight adjusted based on the activity of individual alkaloids) in Beimu extracts significantly correlated with their tracheobronchial relaxation effects (r(2) > 0.9, p < 0.001). CONCLUSIONS: The results demonstrated that the differences in chemical profile of major bioactive isosteroidal alkaloids and pharmacological activity of Beimu could be incorporated into a simple and unified method for quality control and potential prediction of activity of Beimu herbs from different origins.

Comparison and evaluation of antimuscarinic and anti-inflammatory effects of five Bulbus fritillariae species based on UPLC-Q/TOF integrated dual-luciferase reporter assay, PCA and ANN analysis.[Pubmed:28012380]

J Chromatogr B Analyt Technol Biomed Life Sci. 2017 Jan 15;1041-1042:60-69.

Many species of Bulbus fritillariae are used as traditional medicines for thousands of years; however, their application is not standardized. To clarify the differences and homologies, the antimuscarinic and anti-inflammatory effects of five BM species were firstly tested and compared at cellular level. With an integrated strategy combining UPLC-Q/TOF MS, PCA and ANN analysis, the active ingredients among 28 different chemical markers were predicted and identified. SB and QB extracts showed the best antimuscarinic effects and several steroidal alkaloids, such as solanidine, contributed to this effects. However, ZB was superior to reduce the inflammatory response. Another five components were responsible by decreasing the expression of NF-kappaB, including puqiedine, zhepeiresinol, 2-monopalmitin, N-demethylpuqietinone, and Isoverticine. More novelty, a new cluster of five BM species based on active ingredients as potential quality markers was depicted to illustrate their functions. These results of the study could make a reference for the medicinal application of BM species in clinic; and the integrated strategy provided an effective method to obtain the quality markers from medical herbs, which was helpful for the quality control of traditional medicinal products.

Characterization of the Isosteroidal Alkaloid Chuanbeinone from Bulbus of Fritillaria pallidiflora as Novel Antitumor Agent In Vitro and In Vivo.[Pubmed:26584458]

Planta Med. 2016 Feb;82(3):195-204.

Bulbus of Fritillaria pallidiflora, the dried bulb of F. pallidiflora, is widely used in Chinese folk medicine due to its powerful biological activities. The aim of this study was to investigate in vitro and in vivo antitumor activity of different fractions and isosteroidal alkaloids from bulbus of F. pallidiflora and clarify its putative mechanism of antitumor activity. Firstly, we assayed in vitro antitumor effects of different fractions from bulbus of F. pallidiflora and found that chloroform extracts and purified total alkaloids of bulbus of F. pallidiflora showed higher cytotoxic activity than other tested extracts. We further isolated four main alkaloids, chuanbeinone, imperialine-beta-N-oxide, Isoverticine and Isoverticine-beta-N-oxide, from the total alkaloids of bulbus of F. pallidiflora and found that they display significant cytotoxicity, whereby chuanbeinone showed the highest activity against Lewis lung carcinoma cells. Moreover, we found that chuanbeinone induced S phase arrest and further increased apoptosis of Lewis lung carcinoma cells. The results of Western blotting experiments showed that the expression of the antiapoptotic Bcl-2 was reduced by chuanbeinone treatment, while the proapoptotic protein Bax and caspase-3 were increased. Moreover, we investigated the in vivo antitumor activity of chuanbeinone and characterized its putative antitumor mechanism of action by the TUNEL assay and by histological and immunohistochemical analyses. Our results showed that chuanbeinone exhibited significant antitumor activity in vivo, while notably inhibiting tumor angiogenesis and inducing apoptosis characterized by an increased expression of caspase-3. Our findings show that chuanbeinone exhibits significant antitumor activity in vitro and in vivo, and points to possible therapeutic potential for this compound as well as for its natural source, bulbus of F. pallidiflora.

Antitussive, expectorant and anti-inflammatory activities of four alkaloids isolated from Bulbus of Fritillaria wabuensis.[Pubmed:22101082]

J Ethnopharmacol. 2012 Jan 6;139(1):189-93.

ETHNOPHARMACOLOGICAL RELEVANCE: Bulbus Fritillaria Cirrhosae (BFC), known by the Chinese name "Chuan-Bei-Mu", is used as an antitussive, antiasthmatic and expectorant drug for more than 2000 years in China, and Bulbus of Fritillaria wabuensis S. Y. Tang & S. C. Yueh (BFW) was recorded in the 2010 edition of China Pharmacopoeia as one of sources for BFC. The primary objective of this study was to evaluate the antitussive, expectorant and anti-inflammatory effects of alkaloids - imperialine, imperialine-beta-N-oxide, Isoverticine, and Isoverticine-beta-N-oxide, which were isolated from BFW, and to provide experimental evidence for its traditional use. MATERIALS AND METHODS: The alkaloids were isolated using phytochemical methods, and their structures were determined by spectroscopic analysis. Their antitussive effects were measured using murine model of ammonia induced cough, the expectorant effects were evaluated by measuring mice's tracheal phenol red output, and the anti-inflammatory effects were assessed by using the murine model of xylene induced ear edema. RESULTS: The structures of the four alkaloids - imperialine, imperialine-beta-N-oxide, Isoverticine, and Isoverticine-beta-N-oxide isolated from BFW were confirmed. The four alkaloids significantly inhibited cough frequency and increased latent period of cough in mice induced by ammonia. Imperialine and Isoverticine showed obviously antitussive activities in a dose-dependent manner. Besides, the four alkaloids markedly enhanced mice's tracheal phenol red output in expectorant assessment and significantly inhibited the development of ear edema in anti-inflammatory evaluation assay. Moreover, significant differences were found between the structure-activity relationships of the four alkaloids. CONCLUSIONS: The four alkaloids exhibited significant antitussive, expectorant and anti-inflammatory activities. We suggest that they may be the active ingredients of BFW. The results of the present study provided evidence for BFW to be used as an antitussive and expectorant Traditional Chinese Medicine (TCM).

Turbulent-flow chromatography coupled on-line to fast high-performance liquid chromatography and mass spectrometry for simultaneous determination of verticine, verticinone and isoverticine in rat plasma.[Pubmed:20060789]

J Chromatogr B Analyt Technol Biomed Life Sci. 2010 Feb 1;878(3-4):435-41.

A method based on the on-line turbulent-flow chromatography and fast high-performance liquid chromatography/mass spectrometry (TFC-LC/MS) was developed for sensitive and high throughput pharmacokinetic study of traditional Chinese medicines (TCMs). In this method, an on-line extraction column (Waters Oasis HLB) and a fast HPLC column with sub-2 microm particle size (Agilent Zorbax StableBond-C(18), 4.6 mm x 50 mm, 1.8 microm) in a column-switching set-up were utilized. HLB is a reversed-phase extraction column with hydrophilic-lipophilic balanced copolymer (2.1 mm x 20 mm, 25 microm particle size), which will exhibit some turbulent-flow properties at a high-flow rate. The method combines the speed and robustness of turbulent-flow extraction and the sensitivity and separation efficiency of fast HPLC-MS to analyze multiple and trace constituents of TCMs in plasma matrix. This method was successfully applied for pharmacokinetic study of verticine, verticinone and Isoverticine, the chemical markers of Fritillaria thunbergii, after oral administration of total steroidal alkaloids extract of F. thunbergii to rats. Each plasma sample was analyzed within 7 min. The method demonstrated good linearity (R>0.999) ranged from 0.505 to 96.0 ng/mL with satisfactory accuracy and precision, and the lower limit of quantifications of verticine, verticinone and Isoverticine were estimated to be 0.120, 0.595 and 0.505 ng/mL, respectively. These results indicate that the proposed method is fast, sensitive, and feasible for pharmacokinetic study of TCMs.

Isolation and structure elucidation of alkaloids from the bulb of Fritillaria wabuensis.[Pubmed:17253263]

Planta Med. 1998 Jun;64(5):448-50.

Eight alkaloids were isolated from the bulb of Fritillaria wabuensis S. Y. Tang et S. C. Yueh (Liliaceae), which is efficacious against cough. Five of them are known and identified as imperialine ( 1), peimisine ( 2), ebeinone ( 3), Isoverticine ( 4), and imperialine beta- N-oxide ( 5), respectively. Alkaloid 5 is however, controversial. All the signals for both (1)H- and (13)C-NMR spectra of 5 were assigned unambigously by 2D NMR experiments, and (13)C-NMR assignments previously described in the literature were revised. An X-ray structure of alkaloid 5 is presented.

[Studies on the chemical constituents from culbs of hybridized Bulbus Fritillariae Ussuriensis].[Pubmed:15706870]

Zhongguo Zhong Yao Za Zhi. 2004 Apr;29(4):331-4.

OBJECTIVE: To study the chemical constituents of hybridized Bulbus Fritillariae Ussuriensis. METHOD: The chemical constituents were isolated by silica column chromatography and their structures were identified by physical and chemical eveidences and spectral analysis (IR, 1H-NMR, 13C-NMR, 2D-NMR, MS). RESULT: Seven compounds were obtained and identified as (20S,25S)5alpha, 14alpha, 17beta-cevanine-6beta-hydroxy-3-one (hupehenirine, ZF1), (20S,25S)5alpha, 14alpha, 17beta-cevanine-3beta-hydroxy-6-one (hupehenizine, ZF2), (20R,25S)5alpha, 14alpha-cevanine-3beta,20beta-dihydroxy-6-one (peiminine, verticinone, ZF3), (20S,25S)5alpha, 14alpha, 17beta-cevanine-3beta, 6beta-dihydroxy (hupehenine, ZF4), (20R,25S)5alpha, 14alpha-cevanine-3beta, 6beta, 20beta-trihydroxy (Isoverticine, ZF5), (20R,25S)5alpha, 14alpha-cevanine-3beta, 6alpha, 20beta-trihydroxy (peimine, verticine, ZF6), (20S,25S)5alpha, 14alpha, 17beta-evanine-6beta-hydroxy-3beta-O-beta-D-glucoside (hupeheninoside, ZF7). CONCLUSION: Compounds ZF1-7 were isolated from hybridized Bulbus Fritillariae Ussuriensis for the first time.

[A new steroidal alkaloid from the bulbus of Fritillaria wabuensia].[Pubmed:15338876]

Yao Xue Xue Bao. 2004 May;39(5):348-50.

AIM: To investigate the chemical constituents of the bulbs of Fritillaria wabuensia. METHODS: Chromatography techniques were used to isolate the chemical constituents. EI-MS, 1HNMR, 13 CNMR and DEPT were used to determine the structures of the isolated constituents. RESULTS: Three alkaloids were isolated from the bulbs of Fritillaria wabuensia, and were identified as imperialine (I), imperialine-beta-N-oxside (II), Isoverticine-beta-N-oxide (III). CONCLUSION: Isoverticine-beta-N-oxide was isolated from the bulbs of Fritillaria wabuensia for the first time. Isoverticine-beta-N-oxide is a new alkaloid.

Determination of the major isosteroidal alkaloids in bulbs of Fritillaria by high-performance liquid chromatography coupled with evaporative light scattering detection.[Pubmed:11269520]

J Chromatogr A. 2001 Feb 16;909(2):207-14.

A new direct HPLC analytical method using evaporative light scattering detection coupled with a low-temperature adapter for the simultaneous determination of the major biologically active isosteroidal alkaloids in Bulbus Fritillariae, a commonly used antitussive traditional Chinese medicinal (TCM) herb, has been developed. The simultaneous separation of eight Fritillaria alkaloids was achieved on a reversed-phase C8 column with an isocratic mobile phase system consisting of acetonitrile-methanol-water (66.5:3.5:30, v/v) containing 0.006% triethylamine. This method provides good reproducibility and sensitivity for the quantification of six major isosteroidal alkaloids, namely peimissine, verticine, verticinone, imperialine, Isoverticine and ebeiedine in different Fritillaria species with overall intra- and inter-day precision and accuracy of less than 11% and higher than 90%, respectively. The assay was successfully utilized to quantify the major biologically active alkaloids in five Fritillaria species. The results demonstrate that this method is simple, selective, and suitable for the quality control of this commonly used antitussive TCM herb, Bulbus Fritillariae. reserved.

Simultaneous determination of seven major isosteroidal alkaloids in bulbs of Fritillaria by gas chromatography.[Pubmed:10757299]

J Chromatogr A. 2000 Mar 24;873(2):221-8.

The present paper describes the development of a most simple, sensitive, and specific gas chromatographic method to date, for the direct determination of seven major bioactive isosteroidal alkaloids, namely ebeiedine, ebeiedinone, ebeienine, hupehenine, Isoverticine, verticine, verticinone and imperialine, in Fritillaria species, a commonly used antitussive traditional Chinese medicinal (TCM) herb. In the present study, a commercially available Supelco SAC-5 capillary column (30 m x 0.25 mm, 0.25 microm) specifically designed for the analysis of steroids was utilized for the direct determination of Fritillaria alkaloids. Calibration curves were obtained by spiking authentic compounds and the internal standard (solanidine) into herbal samples prior to extraction. Extraction was conducted simply by shaking the pre-alkalized diethyl ether solution (5.0 ml) containing dried herb (0.1 g) for 2 h. All calibration curves showed good linear regressions (r2>0.995) within test ranges. The assay was reproducible and accurate with the overall intra- and inter-day variation and accuracy of less than 10% and more than 90%, respectively. The developed GC method was successfully utilized to analyze seven major bioactive alkaloids in seven Fritillaria species, and the results demonstrate that this direct GC analytical method is suitable for the quality control of this commonly used antitussive TCM herb.

Prederivatization and high-performance liquid chromatographic analysis of alkaloids of bulbs of Fritillaria.[Pubmed:8923321]

J Pharm Sci. 1996 Nov;85(11):1174-9.

A method of prederivatization and HPLC with UV detection was developed for the simultaneous analysis of five major steroidal alkaloids of the Fritillaria species: verticine, verticinone, Isoverticine, ebeiedine, and ebeiedinone. Derivatization was carried out by esterification of the hydroxyl-containing Fritillaria alkaloids to the corresponding naphthoates with 1-naphthoyl chloride. Reaction conditions were optimized and the yields of the derivatization were between 94 and 100% for all test alkaloids. Derivatized alkaloids were characterized by mass spectrometry and HPLC-MS. The validated HPLC-UV method demonstrated linear UV response at the sampling ranges used, and a test limit of 1 microgram/mL was determined for all analytes. This analytical method is simple, convenient, and readily reproducible. The developed method was applied to the analysis of the major pharmacologically active alkaloids in three medicinally used Fritillaria species: F. cirrhosa, F. thunbergii, and F. hupehensis. Five major Fritillaria alkaloids were simultaneously analyzed qualitatively and quantitatively from crude extracts of each of these herbs.

[Isolation and structure flucidation of alkaloids from the bulb of Fritillaria Wabuensis S.Y. Tang et S.C. Yueh].[Pubmed:9208633]

Hua Xi Yi Ke Da Xue Xue Bao. 1996 Mar;27(1):100-5.

Eight alkaloids were isolated from the bulb of Fritillaria Wabuensis S.Y. Tang et S.C. Yueh the best "Chuan Bei" cultivated in Sichuan province, which is efficacious against cough. Five of them are known, identified as imperialine (I), peimisine (II), ebeinone (III), Isoverticine (IV), and imperialine-beta-N-oxide (V). All the signals for both 1H and 12C-NMR spectra of base (V) were assigned by 2D NMR experiments, and an X-ray structural data of alkaloid (V) was obtained for the first time.

[Studies on the chemical constituents of Fritillaria taipaiensis L].[Pubmed:8285053]

Yao Xue Xue Bao. 1993;28(7):516-21.

A new C-nor-D-homo-steroidal alkaloid named taipaienine, together with five known alkaloids namely chuanbeinone, imperialine, verticinone, perimissine and Isoverticine were isolated from the bulbs of Fritillaria taipaiensis L. var. ningxiaensis Y. et W.. The unusual structure of taipaienine was a first sample with a hydroxyl group substituted at C-25 of (22)R-trans quinolizidine moiety with orientation of nitrogen lone pair. Their structures were determined by various spectral analyses.