Chinese Journal of Natural Chinese Journal of Natural Medicines 2013, 11(4): 03300344 Medicines doi: 10.3724/SP.J.1009.2013.00330 Phytochemical and biological research of Fritillaria medicinal resources HAO Da-Cheng1*, GU Xiao-Jie 1, XIAO Pei-Gen2*, PENG Yong2 1Biotechnology Institute, School of Environment, Dalian Jiaotong University, Dalian 116028, China; 2Key Laboratory of Bioactive Substances and Resources Utilization of Chinese Herbal Medicine of Ministry of Education, Insti- tute of Medicinal Plant Development, Chinese Academy of Medical Sciences, Beijing 100193, China Available online 20 July 2013 [ABSTRACT] The genus Fritillaria is a botanical source for various pharmaceutically active components, which have been commonly used in traditional Chinese medicine for thousands of years. Increasing interest in Fritillaria medicinal resources has led to additional discoveries of steroidal alkaloids, saponins, terpenoids, glycosides and many other compounds in various Fritillaria species, and to investigations on their chemotaxonomy, molecular phylogeny and pharmacology. In continuation of studies on Fritillaria pharmaco- phylogeny, the phytochemistry, chemotaxonomy, molecular biology and phylogeny of Fritillaria and their relevance to drug efficacy is reviewed. Literature searching is used to characterize the global scientific effort in the flexible technologies being applied. The interre- lationship within Chinese Bei Mu species and between Chinese species, and species distributed outside of China, is clarified by the molecular phylogenetic inferences based on nuclear and chloroplast DNA sequences. The incongruence between chemotaxonomy and molecular phylogeny is revealed and discussed. It is essential to study more species for both the sustainable utilization of Fritillaria medicinal resources and for finding novel compounds with potential clinical utility. Systems biology and omics technologies will play an increasingly important role in future pharmaceutical research involving the bioactive compounds of Fritillaria. [KEY WORDS] Fritillaria; Chemical components; Chemotaxonomy; Molecular taxonomy; Phylogeny; Medicinal resource [CLC Number] R284, R965 [Document code] A [Article ID] 1672-3651(2013)04-0330-015 Zhe Bei Mu are the most famous Chinese herbal medicines 1 Introduction obtained from Fritillaria species. The former is from F. cir- Fritillaria is a genus of about 130−165 species [1-2] rhosa, F. unibracteata, F. przewalskii, F. delavayi, F. tai- within the monocot family Liliaceae, and is native to temper- paiensis, and F. wabuensis, while the latter is from F. thun- ate regions of the Northern Hemisphere. Fritillaria is a bo- bergii and F. thunbergii var. chekiangensis. According to tanical source for various pharmaceutically active compo- traditional descriptions, Fritillaria is slightly cold, and affects nents which have been used in traditional Chinese medicine the lungs (to clear heat and moisten dryness, and used for for thousands of years. Many species (such as F. cirrhosa, F. hot-type bronchitis with dry cough) and the heart (to calm thunbergii and F. verticillata) are used in traditional Chinese heart fire). Fritillaria is also used for treating lumps beneath cough remedies. In the China Pharmacopoeia (http://www. the skin, such as scrofulous swellings and breast lumps. Zhe chp. org.cn/cms/home/) they are listed as Chuan Bei Mu, Zhe Bei Mu is often used for the treatment of lumps, and the Bei Mu, Yi Bei Mu, Hu Bei Bei Mu, and Ping Bei Mu, re- moistening property attributed to Chuan Bei Mu is not need- spectively, and are often found in formulations combined ed. It has been adopted into some Chinese herb formulas for with extracts of Loquat (Eriobotrya japonica). F. verticillata treating cancers. Different Fritillaria species possess bulbs are also traded as Bei Mu in Japan. Chuan Bei Mu and different chemical profiles, and may have different pharma- cological effects. However, it is quite difficult to authenticate [Received on] 29-Oct.-2012 Fritillaria species only by morphology. [Research funding] This project was supported by Ministry of Sci- To date, at least 30 Fritillaria species have been charac- ence and Technology national support program (No. 2012BAI29B01) terized for their chemical components [3]. Advances in ana- [*Corresponding author] HAO Da-Cheng: Prof., E-mail: hao@djtu. edu.cn; XIAO Pei-Gen: Prof., E-mail: [email protected]. net.cn lytical chemistry and molecular biology techniques facilitate These authors have no conflict of interest to declare. the in-depth studies of Fritillaria pharmaceutical resources. 330 Chin J Nat Med Jul. 2013 Vol. 11 No. 4 2013 年 7 月 第 11 卷 第 4 期 HAO Da-Cheng, et al. / Chinese Journal of Natural Medicines 2013, 11(4): 330344 However, it is essential to study more species for both the NMR spectroscopy and MS [5]. The new cevanine-type alka- sustainable utilization of Fritillaria medicinal resources and loids, (3β, 5α)-20-hydroxy-6-oxocevan-3-yl acetate 7 and (3β, for finding novel compounds with potential clinical utility. In 5α, 6α)-6, 20-dihydroxycevan-3-yl acetate 8, were isolated this brief review, we focus on recent progress in the phyto- from bulbs of F. hupehensis and characterized by spectral chemistry and chemotaxonomy of Fritillaria, as well as mo- analysis [6]. Bulbs of F. unibracteata in Sichuan, China, lecular taxonomy and phylogeny. To date, very few studies contained the new glycoalkaloid puqiedinone-3-O-β-D- have attempted to correlate DNA based phylogeny of me- glucopyranoside 11, whose structure was elucidated by spec- dicinal plants with phytochemistry or with medicinal proper- troscopic and chemical methods [7]. Phytochemical investiga- [4] ties . Here the molecular phylogeny of world-wide Fritil- tion of the bulbs of F. lichuanensis yielded a new isosteroidal laria species is reconstructed and the results compared with alkaloid hupehenizioiside 13 [8]. A further investigation of those of chemotaxonomy. Phylogeny has great explanatory bulbs of F. lichuanensis resulted in the isolation of two power and offers a unique perspective to complement unique cevanine-type alkaloids, lichuanine ((20S, 25R)-5α, chemotaxonomy. 14α-cevanine-3β, 6β-diol) 14 and lichuanisinine ((20S, [9] 2 Chemical Components of the Genus Fritillaria 25S)-5α, 14α-cevanine-3β, 6β-diol-N-oxide) 15 . Extraction of the bulbs of F. puqiensis yielded three new veratra- 2.1 Steroidal alkaloids mine-type alkaloids, namely puqienine C 3, puqienine D 4 Alkaloids are a group of naturally occurring chemical and puqienine E 5 [5]. Alkaloids 9 and 10 were obtained from compounds that contain mostly basic nitrogen atoms. The F. hupehensis. Their structures were deduced to be (3β, 5α, steroidal alkaloids from Fritillaria can be classified into 13α, 23β)-7, 8, 12, 14-tetradehydro-5, 6, 12, 13-tetrahydro-3, two groups based on the carbon framework: isosteroidal 23-dihydroxyveratraman-6-one and (3β, 5α, 13α, 23β)-7, 8, alkaloids and steroidal alkaloids (Fig. 1). According to the 12, 14-tetradehydro-5, 6, 12, 13-tetrahydro-3, 13, 23- trihy- linkage patterns between rings E and F, isosteroidal alka- droxy-veratraman-6-one, respectively, on the basis of chemi- loids can be divided into cevanine (A), jervine (B) and cal shift comparisons and 1H, 1H-COSY, HMBC correlations veratramine (C) types (Figs. 1 and 2). Steroidal alkaloids [6]. Bulbs of F. ussuriensis have yielded a new alkaloid ping- can be divided into solanidine (D) and secosolanidine (E) beimunone A 16 with an aromatized D-ring, whose structure [2] types , depending on whether the nitrogen atom is incor- was elucidated on the basis of spectral analysis. Chemical porated into an indolizidine ring or a piperidine ring (Fig. 1). investigation of the bulbs of F. puqiensis afforded puqienine Steroidal alkaloids found in Fritillaria before 2005 have F 17. This novel veratramine alkaloid possessed a 12, been summarized [3]. Table 1 summarizes the steroidal al- 16-epoxy ring [10]. Peimisine-3-O-β-D-glucopyranoside 12, kaloids found since 2006. possessing the furan ring E fused onto a piperidine ring sys- Puqiedine 1 and 3α-puqiedin-7-ol 2 were isolated from F. tem forming an ether bridge between carbon atoms C17 and puqiensis, their structures being determined on the basis of C23, was isolated from F. unibracteata, which might be identical Fig. 1 Types of steroidal alkaloids in Fritillaria. a, isosteroidal alkaloids; b, steroidal alkaloids. Refer to Table 1 for more details 2013 年 7 月 第 11 卷 第 4 期 Chin J Nat Med Jul. 2013 Vol. 11 No. 4 331 HAO Da-Cheng, et al. / Chinese Journal of Natural Medicines 2013, 11(4): 330344 Fig. 2 Steroidal alkaloids of Fritillaria. D/E configurations of cevanine group steroidal alkaloids of 1, 2, 7, 8, 11, 13−15 are trans-form. Refer to Table 1 for more details with the known jervine-type compound peimisine on the induced contraction of guinea-pig tracheal rings [14]. The basis of NMR data [7]. Pengbeimine B 18 and pengbeimine D relaxant action of imperialine and sinpeinine A, the cis-D/E 19 from F. mo na t h a have a novel skeleton in that C13 and cevanine alkaloids, is due to their selective inhibitory effects C22 were connected through oxygen forming a furan ring. on muscarinic M2 receptors. Verticine, verticinone, imperi- Their absolute structures were established by X-ray diffrac- aline, imperialine-3beta-D-glucoside and puqietinone, i.e., tion [11]. both trans- and cis-D/E cevanine alkaloids, significantly An extract of the dried ground bulbs of F. puqiensis af- elevate the cAMP concentration in the HEK cells transfected forded the verazine-type alkaloid puqietinedione 6. Its struc- with muscarinic M(2) receptor plasmid [15]. N-Demethylpu- ture was formulated as (22R, 25S)-N-methyl-22, 26- epimi- qietinone, hupeheninoside, ebeiedinone, yibeinoside A, and nocholest-3, 6-dione by comparison of the NMR data with chuanbeinone inhibited the bioactivity of human whole blood those of puqietinone [5]. Delavidine 20, an unusual 6, 12, acetylcholinesterase and plasma butyrylcholinesterase [16]. 22-triketo verazine type steroidal alkaloid with a pyrrolidine Imperialine, chuanbeinone, verticinone, and verticine from F.