Two New Flavonol Glycosides and a Metabolite Profile of Bryophyllum pinnatum, a Phytotherapeutic Used in Obstetrics and Gynaecology

Abstract

Bryophyllum pinnatum is a succulent perennial plant native to Madagascar which is used in anthroposophical medicine to treat psychiatric disorders and as a tocolytic agent to prevent premature labour. We performed a metabolite profiling study in order to obtain a comprehensive picture of the constituents in B. pinnatum leaves and to identify chromatographic markers for quality control and safety assessment of medicinal preparations. Preliminary HPLC-PDA-ESIMS analyses revealed that flavonoid glycosides were the main UV-absorbing constituents in the MeOH extract of B. pinnatum. Two phenolic glucosides, syringic acid β-D-glucopyranosyl ester (1) and 4′-O-β-D-glucopyranosyl-cis-p-coumaric acid (2), as well as nine flavonoids (3-11) including kaempferol, quercetin, myricetin, acacetin, and diosmetin glycosides were unambiguously identified by ¹H and 2D NMR analysis after isolation from a MeOH extract. The flavonol glycosides quercetin 3-O-α-L-arabinopyranosyl-(1 → 2)-α-L-rhamnopyranoside 7-O-β-D-glucopyranoside (3) and myricetin 3-O-α-L-arabinopyranosyl-(1 → 2)-α-L-rhamnopyranoside (4) were new natural products. With the aid of HPLC-PDA-APCIMS and authentic references isolated from the related species B. daigremontianum, the presence of four bufadienolides, bersaldegenin-1-acetate (12), bryophyllin A (13), bersaldegenin-3-acetate (14), and bersaldegenin-1,3,5-orthoacetate (15) was detected in B. pinnatum.

Key words

Bryophyllum pinnatum - Kalanchoe pinnata - Crassulaceae - HPLC-PDA-MS profiling - flavonoids - bufadienolides

• References

• 1 Yadav NP, Dixit VK. Hepatoprotective activity of leaves of Kalanchoe pinnata Pers. J Ethnopharmacol 2003; 86: 197-202
• 2 Ajibesin KK. Ethnobotanical survey of plants used for skin diseases and related ailments in Akwa Ibom State, Nigeria. Ethnobot Res Appl 2012; 10: 463-522
• 3 Ghasi S, Egwuibe C, Achukwu PU, Onyeanusi JC. Assessment of the medical benefit in the folkloric use of Bryophyllum pinnatum leaf among the Igbos of Nigeria for the treatment of hypertension. Afr J Pharm Pharmacol 2011; 5: 83-92
• 4 Lans CA. Ethnomedicines used in Trinidad and Tobago for urinary problems and diabetes mellitus. J Ethnobiol Ethnomed 2006; 2: 45
• 5 Daems W, Kaufmann H, Titze O. Kurzgefasste Bryophyllum-Chronologie. Weleda Korrespondenzblätter für Ärzte 1982; 105: 5-11
• 6 Hassauer W, Schreiber K, von der Decken D. Bryophyllum – Ein neuer Weg in der tokolytischen Therapie. Erfahrungsheilkunde 1985; 34: 683-687
• 7 Plangger N, Rist L, Zimmermann R, von Mandach U. Intravenous tocolysis with Bryophyllum pinnatum is better tolerated than beta-agonist application. Eur J Obstet Gynecol Reprod Biol 2006; 124: 168-172
• 8 Simões-Wüst AP, Grãos M, Duarte CB, Brenneisen R, Hamburger M, Mennet M, Ramos MH, Schnelle M, Wächter R, Worel AM, von Mandach U. Juice of Bryophyllum pinnatum (Lam.) inhibits oxytocin-induced increase of the intracellular calcium concentration in human myometrial cells. Phytomedicine 2010; 17: 980-986
• 9 Wächter R, Brenneisen R, Hamburger M, Mennet M, Schnelle M, Worel AM, Simoes-Wust AP, von Mandach U. Leaf press juice from Bryophyllum pinnatum (Lamarck) Oken induces myometrial relaxation. Phytomedicine 2011; 19: 74-82
• 10 Schuler V, Suter K, Fürer K, Eberli D, Horst M, Betschart C, Brenneisen R, Hamburger M, Mennet M, Schnelle M, Simoes-Wust AP, von Mandach U. Bryophyllum pinnatum inhibits detrusor contractility in porcine bladder strips – a pharmacological study towards a new treatment option of overactive bladder. Phytomedicine 2012; 19: 947-951
• 11 Betschart C, von Mandach U, Seifert B, Scheiner D, Perucchini D, Fink D, Geissbuhler V. Randomized, double-blind placebo-controlled trial with Bryophyllum pinnatum versus placebo for the treatment of overactive bladder in postmenopausal women. Phytomedicine 2013; 20: 351-358
• 12 Muzitano MF, Tinoco LW, Guette C, Kaiser CR, Rossi-Bergmann B, Costa SS. The antileishmanial activity assessment of unusual flavonoids from Kalanchoe pinnata . Phytochemistry 2006; 67: 2071-2077
• 13 Pal S, Nag Chaudhuri AK. Studies on the anti-ulcer activity of a Bryophyllum pinnatum leaf extract in experimental animals. J Ethnopharmacol 1991; 33: 97-102
• 14 Ofokansi K, Esimone C, Anele C. Evaluation of the in vitro combined antibacterial effect of the leaf extracts of Bryophyllum pinnatum (Fam. Crassulaceae) and Ocimum gratissimum (Fam. Labiatae). Plant Prod Res J 2005; 9: 23-27
• 15 Akinpelu DA. Antimicrobial activity of Bryophyllum pinnatum leaves. Fitoterapia 2000; 71: 193-194
• 16 Supratman U, Fujita T, Akiyama K, Hayashi H, Murakami A, Sakai H, Koshimizu K, Ohigashi H. Anti-tumor promoting activity of bufadienolides from Kalanchoe pinnata and K. daigremontiana x tubiflora . Biosci Biotechnol Biochem 2001; 65: 947-949
• 17 Almeida AP, Da Silva SA, Souza ML, Lima LM, Rossi-Bergmann B, de Moraes VL, Costa SS. Isolation and chemical analysis of a fatty acid fraction of Kalanchoe pinnata with a potent lymphocyte suppressive activity. Planta Med 2000; 66: 134-137
• 18 Ojewole J. Antinociceptive, anti-inflammatory and antidiabetic effects of Bryophyllum pinnatum (Crassulaceae) leaf aqueous extract. J Ethnopharmacol 2005; 99: 13-19
• 19 Kamboj A, Saluja A. Bryophyllum pinnatum (Lam.) Kurz.: Phytochemical and pharmacological profile: A Review. Pharmacogn Rev 2009; 3: 364-374
• 20 Muzitano MF, Cruz EA, de Almeida AP, Da Silva SA, Kaiser CR, Guette C, Rossi-Bergmann B, Costa SS. Quercitrin: an antileishmanial flavonoid glycoside from Kalanchoe pinnata . Planta Med 2006; 72: 81-83
• 21 Yamagishi T, Haruna M, Yan XZ, Chang JJ, Lee KH. Antitumor agents, 110. Bryophyllin B, a novel potent cytotoxic bufadienolide from Bryophyllum pinnatum . J Nat Prod 1989; 52: 1071-1079
• 22 Wagner H, Lotter H, Fischer M. The toxic and sedative bufadienolides of Kalanchoe daigremontiana HAMET et PERR. Helv Chim Acta 1986; 69: 359-367
• 23 Muzitano MF, Bergonzi MC, De Melo GO, Lage CLS, Bilia AR, Vincieri FF, Rossi-Bergmann B, Costa SS. Influence of cultivation conditions, season of collection and extraction method on the content of antileishmanial flavonoids from Kalanchoe pinnata . J Ethnopharmacol 2011; 133: 132-137
• 24 El Abdellaoui S, Destandau E, Toribio A, Elfakir C, Lafosse M, Renimel I, Andre P, Cancellieri P, Landemarre L. Bioactive molecules in Kalanchoe pinnata leaves: extraction, purification, and identification. Anal Bioanal Chem 2010; 398: 1329-1338
• 25 Inoshiri S, Sasaki M, Kohda H, Otsuka H, Yamasaki K. Aromatic glycosides from Berchemia racemosa . Phytochemistry 1987; 26: 2811-2814
• 26 Rasmussen S, Wolff C, Rudolph H. 4′-O-beta-D-glucosyl-cis-p-coumaric acid – a natural constituent of Sphagnum fallax cultivated in bioreactors. Phytochemistry 1996; 42: 81-87
• 27 Zhong XN, Otsuka H, Ide T, Hirata E, Takushi A, Takeda Y. Three flavonol glycosides from leaves of Myrsine seguinii . Phytochemistry 1997; 46: 943-946
• 28 Soicke H, Gorler K, Waring H. Flavonol glycosides from Moghania faginea . Planta Med 1990; 56: 410-412
• 29 Neretina OV, Fedorov SV, Gromova AS, Lutskii VI, Elʼkin YN. Flavonoids of Hedysarum setigerum . Chem Nat Compd 2002; 38: 194-195
• 30 Wu J, Yi W, Jin L, Hu D, Song B. Antiproliferative and cell apoptosis-inducing activities of compounds from Buddleja davidii in Mgc-803 cells. Cell Div 2012; 7: 20
• 31 McKenzie RA, Franke FP, Dunster PJ. The toxicity to cattle and bufadienolide content of six Bryophyllum species. Aust Vet J 1987; 64: 298-301
• 32 Capon RJ, Macleod JK, Oelrichs PB. Bryotoxins B and C, toxic bufadienolide orthoacetates from the flowers of Bryophyllum tubiflorum (Crassulaceae). Aust J Chem 1986; 39: 1711-1715
• 33 Yamagishi T, Yan XZ, Wu RY, McPhail DR, McPhail AT, Lee KH. Structure and stereochemistry of bryophyllin A, a novel potent cytotoxic bufadienolide orthoacetate from Bryophyllum pinnatum . Chem Pharm Bull 1988; 36: 1615-1617
• 34 Supratman U, Fujita T, Akiyama K, Hayashi H. New insecticidal bufadienolide, bryophyllin C, from Kalanchoe pinnata . Biosci Biotechnol Biochem 2000; 64: 1310-1312