Biochemical Systematics and Ecology 86 (2019) 103923
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Chemical constituents from the leaves of Crataegus pinnatifida Bge a
Wanchun Chu , Pinyi Gao
, Lingzhi Li
School of Traditional Chinese Materia Medica, Key Laboratory of Computational Chemistry-Based Natural Antitumor Drug Research & Development, Liaoning Province, Shenyang Pharmaceutical University, Shenyang, 110016, China College of Pharmaceutical and Biotechnology Engineering, Shenyang University of Chemical Technology, Shenyang, 110142, China
Keywords: Rosaceae Crataegus pinnatifida Triterpenoids Flavonoids Chemotaxonomy
The phytochemical investigations on the leaves of Crataegus pinnatifida led to the isolation of 20 compounds, including seven triterpenoids (1-7), three hydroxycinnamic acids (8-10), three lignans (11-13) and seven flavonoids (14-20). All chemical structures were established on the basis of NMR (1H NMR, 13C NMR) spectroscopic data. Meanwhile, compounds 3-12 are reported for the first time from Crataegus genus. In addition, compounds 10-11 are isolated from the family Rosaceae for the first time. On the basis of chemical research, the chemotaxonomic significance of the isolated compounds has been discussed.
1. Subject and source
3. Present work
Crataegus pinnatifida Bge. (also known as hawthorn), belonging to Crataegus genus and family Rosaceae, is widely distributed in northern temperate zones, including those of Eastern Asia, Europe, Central Asia and North America (Song et al., 2012). The fruits and leaves of C. pinnatifida are used in Chinese traditional medicine for the treatment of postpartum blood stasis, cardiodynia and hyperlipoidemia. Moreover, the leaves are also used to cure dyspepsia (Chinese Pharmacopoeia Commission, 2015). The leaves of C. pinnatifida were purchased from Anguo County of Hebei Province, China in April 2015, and identified by Prof. Jin-Cai Lu, College of Traditional Chinese Medicines, Shenyang Pharmaceutical University. A voucher specimen (No. SZ150405) is kept in the Nature Products Laboratory of Shenyang Pharmaceutical University, Shenyang, PR China.
Air-dried and powdered leaves of C. pinnatifida (10 kg) were extracted with 80% EtOH to yield an extract (2 kg, three times, each time 2 h). And, the crude extract was separated by the macroporous resin chromatography with EtOH/H2O (0:100 → 100:0, v/v). The 55% ethanol fraction (300 g) was subsequently separated by silica gel column chromatography and eluted with CH2Cl2/MeOH (20:1, 10:1, 8:1, 6:1, 5:1, 3:1, 1:1, 0:1, v/v) to afford seven fractions (Frs.1–7) based on TLC profiles. Among which Fr.2 was separated by silica gel CC (CH2Cl2/MeOH 20:1 → 8:1) to provide four parts (Frs.2.1–2.4). Fr.2.2 was subjected to MCI-CHP-20p column chromatography eluting with MeOH/H2O (0:100 → 100:0) and was further purified by high performance liquid chromatography resulted in the isolation of compounds 1 (23 mg), 2 (9 mg), 3 (15 mg), 4 (12 mg), 5 (25 mg), 6 (25 mg), 7 (13 mg), 8 (11 mg) and 20 (17 mg). Fr.3 was purified by CC (SiO2; CH2Cl2/MeOH 10:1 → 6:1), sequentially by CC (CHP-20p; MeOH/H2O 0:100 → 100:0) and Sephadex LH-20 column chromatography to yield compounds 14 (13 mg), 15 (12 mg), 16 (15 mg), 17 (17 mg), 18 (36 mg) and 19 (59 mg), Fr.4 was subjected to silica gel CC eluting with CH2Cl2/MeOH gradient system (8:1 → 5:1, v/v) to obtain five fractions (Fr.4.1-Fr.4.5). Separation of Fr.4.4 on a CHP-20p CC and HPLC afforded compounds 9 (17 mg), 10 (12 mg), 11 (13 mg), 12 (14 mg) and 13 (12 mg). Chemical structure elucidation of these compounds (Fig. 1) was achieved by the analysis of their spectroscopic (1H NMR, 13C NMR) and
2. Previous work Previously, more than 180 compositions have been identified from various parts of C. pinnatifida (stems, leaves, flowers, fruits and seeds) (Wu et al., 2014; Guo et al., 2018; Huang et al., 2018), including flavonoids (Li et al., 2015), triterpenoids (Song et al., 2011), monoterpenoids (Li et al., 2015), sesquiterpenoids (Zhou et al., 2014), lignans (Huang et al., 2015), hydroxycinnamic acids, steroids and nitrogen-containing compounds (Wu et al., 2014).
Corresponding author. Corresponding author. College of Pharmaceutical and Biotechnology Engineering, Shenyang University of Chemical Technology, Shenyang, 110142, China E-mail address: [email protected]
https://doi.org/10.1016/j.bse.2019.103923 Received 31 January 2019; Received in revised form 3 July 2019; Accepted 13 July 2019 0305-1978/ © 2019 Elsevier Ltd. All rights reserved.
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Fig. 1. Chemical structures of compounds 1–20.
comparison with those reported data in the related literature. They were identified as oleanolic acid (1) (Seebacher et al., 2003), ursolic acid (2) (Gnoatto et al., 2008), crataegolic acid (3) (Taniguchi et al., 2002), arjungenin (4), arjunglucoside (5) (Jossang et al., 1996), 2α,3α,19α-trihydroxyurs-12-en-28-oicacid (6), tormentic acid-28-O-βD-glucopyrannoside (7) (Liu et al., 2014), eugenyl-O-β-D-glucopyranoside (8) (Yoshio Takeda et al., 1998), sasanquin (9) (Zhao et al., 1999), eugenyl-O-β-D-apiofuranosyl-(1 → 6)-O-β-D-glucopyranoside (10) (Takeda et al., 1998), (+)-lariciresinol-4-O-β-D-glucopyranoside (11), (+)-pinoresionol-4′-O-β-D-glucopyranoside (12) (Huang and Kong, 2003), tortoside A (episyringaresinol-4″-O-β-D-monoglucopyranoside) (13) (Cai et al., 2004), kaempferol (14) (Ding et al., 2013), quercetin (15) (Wang et al., 2008), 3-O-β-D-galacopyanosyl quercetin (16) (Su et al., 2018), rutin (17) (Xiao et al., 2005), vitexin (18) (Zhang and Xu, 2003), 2″-O-rhamnosyl vitexin (19) (Xu et al., 2009) and eriodectyol (20) (Li et al., 2015).
taxonomic characters. Rosaceae family comprises four subfamily and approximately 124 genera. In addition, phylogenic and morphologic analysis suggested that Crataegus with 3300 species, is a polyphyletic genus. Most genera exhibit morphological diversity, which can result in some confusions in the identification of a species. Further research is needed to enrich our knowledge of the relationships among these genera. The family Rosaceae is characterized by the occurrence of triterpenoids. To our knowledge, the most common are ursolic acid-type and oleanolic acid-type triterpenoids. Compounds 1 and 3–5 are oleanolic acid-type and compounds 2 and 6–7 are ursolic acid-type triterpenoids. Moreover, triterpenoids (1–7) isolated at this work have been found in the genera of subfamily Maloideae Weber and Rosoideae Focke sources, which suggested the wide distribution of triterpenoids in Rosaceae family, particularly in Rosoideae Focke subfamily (Fig. 2). Interestingly, most of these compounds possess a 19α-hydroxyl. Thus, they could be useful taxonomic markers for the identification of this family. Moreover, this study has shown that the triterpenoid glycosides isolated from C. pinnatifida all contain a 28-O-glycosides. Compound 2 is frequently described from species of Crataegus and is officially listed in the Chinese Pharmacopoeia (2015 edition) as a compound that can be used for the identification of Crataegi Fructus. Compounds 3–7 were isolated from C. pinnatifida for the first time, which extend the knowledge about chemical diversity of C. pinnatifida. In our present research, compounds 3, 4 and 5 occurs sporadically in Rosoideae Focke subfamily and 3 mainly isolated from Maloideae Weber subfamily (Kim et al., 2017; Banno et al., 2005). Compound 4 was also isolated from
4. Chemotaxonomic significance The present work reports the isolation and identification of 20 compounds from the leaves of C. pinnatifida, including seven triterpenoids (1–7), three hydroxycinnamic acids (8–10), three lignans (11–13) and seven flavonoids (14–20). Among the isolates, compounds 3–12 are reported from the genus Crataegus for the first time. In addition, compounds 10–11 are isolated from Rosaceae family for the first time. The chemistry of the Rosaceae is extensive and dominated by reports concerning triterpenoids, phenolics and flavones which may serve as 2
Biochemical Systematics and Ecology 86 (2019) 103923
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Fig. 2. The chemotaxonomic relevance of trepenoids (1–7) and flavones (14–20) for Rosaceae.
both Pyrus and Rubus, which may indicate a close relationship among the three different genera in Rosaceae family (Lia et al., 2015; Zhong et al., 2001). In addition, compound 5 was also obtained from Potentilla and Rubus genus (Morikawa et al., 2014; Zhou et al., 1992). Therefore, it may indicates a close relationship among these three genera from Rosaceae family. And Compounds 6–7 are common ingredients widely distributed in different genus in Rosoideae Focke subfamily. Previous chemical investigations showed that species of Crataegus are rich in flavonoids and their glycosides. In our study, the most abundant flavonoid glycosides in C. pinnatifida are glucoside, galactoside and rhamnoside. The flavonoid glycosides belonging to the family Rosaceae are mainly 3-O-glycosides, and 8-C-glycosides appears less frequently. In addition, it was found that compounds 14–17 are widely distributed in various subfamilies of Rosaceae family (Fig. 2). But to our knowledge, compounds 18 and 19 (8-C-glycosides) were mainly isolated from species of Crataegus rather than from other genus of the family Rosaceae (Sagaradze et al., 2017; Zhang and Xu, 1999) (Fig. 2). These findings support the fact that compounds 18–19 (8-C-glycosides) may be chemo-systematic markers for taxa in the genus Crataegus. On the other hand, the result also showed that the presence of compounds 14–17 (3-O-glycosides) supports the taxonomic position of the genus Crataegus in the Rosaceae family. Compound 20 reported from Crataegus (Li et al., 2015) and Malus only, which indicated a close relationship between Crataegus and Malus of the Rosaceae family. In addition, compound 20, which has a flavanone skeleton, are rarely isolated from C. pinnatifida when compared to flavones and flavonols. Overall, the phytochemical data increases our knowledge about the chemistry of C. pinnatifida.
Cai, X.F., Lee, I.S., Dat, N.T., Shen, G., Kang, J.S., Kim, D.H., Kim, Y.H., 2004. Arch Pharm. Res. (Seoul) 27, 738. Chinese Pharmacopoeia Commission, 2015. Pharmacopoeia of the People's Republicof China, vol. 1. China Medical Science Press, Beijing, pp. 31–32. Ding, H.Y., Lin, H.C., Teng, C.M., Wu, Y.C., 2013. J. Chin. Chem. Soc. 47, 381–388. Gnoatto, S.C., Dassonville-Klimpt, A., Da Nascimento, S., Galera, P., Boumediene, K., Gosmann, G., Sonnet, P., Moslemi, S., 2008. Eur. J. Med. Chem. 43, 1865–1877. https://doi:10.1016/j.ejmech.2007.11.021. Guo, R.L., Lin, B., Shang, X.Y., Zhou, L., Yao, G.D., Huang, X.X., Song, S.J., 2018. Fitoterapia 127, 301–307. https://doi.org/10.1016/j.fitote.2018.03.003. Huang, W.W., Kong, D.Y., 2003. Chin. J. Nat. Med. 1, 199–203. Huang, X.X., Bai, M., Zhou, L., Lou, L.L., Liu, Q.B., Zhang, Y., Li, L.Z., Song, S.J., 2015. J. Agric. Food Chem. 63, 7252–7260. Huang, X.X., Xu, Y., Bai, M., Zhou, L., Song, S.J., Wang, X.B., 2018. Nat. Prod. Res. 32, 1706–1713. https://doi.org/10.1080/14786419.2017.1399378. Jossang, A., Seuleiman, M., Maidou, E., Bodo, B., 1996. Phytochemistry 41, 591–594. Kim, C.S., Subedi, L., Oh, J., Kim, S.Y., Choi, S.U., Lee, K.R., 2017. J. Nat. Prod. 80, 1134–1140. Li, L.-Z., Gao, P.-Y., Song, S.-J., Yuan, Y.-Q., Liu, C.-T., Huang, X.-X., Liu, Q.-B., 2015. J. Funct. Foods 12, 237–245. https://doi.org/10.1016/j.jff.2014.11.012. Lia, Z.-J., Wan, C.-P., Cai, L., Li, S.-Q., Zheng, X., Qi, Y., Dong, J.-W., Yin, T.-P., Zhou, Z.X., Tan, N.-H., Ding, Z.-T., 2015. Phytochem. Lett. 13, 246–251. https://doi.org/10. 1016/j.phytol.2015.06.015. Liu, Y., Cheng, L., He, Q.Q., Yan, Z.H., Kong, D.Y., 2014. Chin. Tradit. Herb. Drugs 45, 2742–2747. Morikawa, T., Ninomiya, K., Imura, K., Yamaguchi, T., Akagi, Y., Yoshikawa, M., Hayakawa, T., Muraoka, O., 2014. Phytochemistry 102, 169–181. https://doi.org/ 10.1016/j.phytochem.2014.03.002. Sagaradze, V.A., Babaeva, E.Y., Kalenikova, E.I., 2017. Pharm. Chem. J. 51, 277–280. Seebacher, W., Simic, N., Weis, R., Saf, R., Kunert, O., 2003. Magn. Reson. Chem. 41, 636–638. Song, S.J., Li, L.Z., Gao, P.Y., Peng, Y., Yang, J.Y., Wu, C.F., 2011. Food Chem. 129, 933–939. Song, S.J., Li, L.Z., Gao, P.Y., Yuan, Y.Q., Wang, R.P., Liu, K.C., Peng, Y., 2012. Planta Med. 78, 1967–1971. Su, X.D., Li, W., Ma, J.Y., Kim, Y.H., 2018. Nat. Prod. Res. 32, 2347–2351. https://doi. org/10.1080/14786419.2017.1405412. Takeda, Y., Ooiso, Y.i, Masuda, T., Honda, G., Otsuka, T., Ekrem, S., Yesilada, E., 1998. Phytochemistry 49, 787–791. Taniguchi, S., Imayoshi, Y., Kobayashi, E., Takamatsu, Y., Ito, H., Hatano, T., Sakagamib, H., Tokudac, H., Nishinoc, H., Sugitad, D., Shimurad, S., Yoshida, T., 2002. Phytochemistry 59, 315–323. Wang, L.L., Liu, B., Shi, R.B., T, G.Z., 2008. J. Beijing Univ. Tradit. Chin. Med. 31, 116–118. Wu, J., Peng, W., Qin, R., Zhou, H., 2014. Molecules 19, 1685–1712. Xiao, Y.M., Wu, Q., Wu, W.B., Zhang, Q.Y., Lin, X.F., 2005. Biotechnol. Lett. 27, 1591–1595. Xu, H., Zhou, T., Wen, J., Fan, G., Wu, Y., 2009. J. Liq. Chromatogr. Relat. Technol. 32, 2216–2231. https://doi.org/10.1080/10826070903163008. Zhang, P.-C., Xu, S.-X., 1999. Chin. J. Med. Chem. 9, 214–215 234. Zhang, P.-C., Xu, S.-X., 2003. J. Asian Nat. Prod. Res. 5, 131–136. http://www. tandfonline.com/loi/ganp20. Zhao, W.M., Qin, G.W., Xu, R.S., Li, X.Y., Liu, J.S., Wang, Y., Feng, M., 1999. Fitoterapia 70, 529–531. Zhong, W.Q., Ding, L.S., Wang, M.K., 2001. Chin. Tradit. Herb. Drugs 32, 874–876. Zhou, X.H., Kasai, R., Ohtani, K., Tanaka, O., Nie, R.L., Yang, C.R., Zhou, J., Yamasaki, k., 1992. Phytochemistry 31, 3642–3644. Zhou, C.C., Huang, X.X., Gao, P.Y., Li, F.F., Li, D.M., Li, L.Z., Song, S.J., 2014. J. Asian Nat. Prod. Res. 16, 169–174. http://www.tandfonline.com/loi/ganp20.
Acknowledgments The authors gratefully thank National Natural Science Foundation of China (81302661), the Scientific Research Starting Foundation for Doctors of Liaoning province of China (20121106) and the Foundation from the Project of Education Department of Liaoning Province of China(L2012358). Appendix A. Supplementary data Supplementary data to this article can be found online at https:// doi.org/10.1016/j.bse.2019.103923. References Banno, N., Akihisa, T., Tokuda, H., Yasukawa, K., Nishino, H., 2005. Biol. Pharm. Bull. 28, 1995–1999.