Polyoxypregnane steroids with an open-chain sugar moiety from Marsdenia tenacissima and their chemoresistance reversal activity

doi:10.1016/j.phytochem.2016.03.006

Phytochemistry

Volume 126, June 2016, Pages 47-58

https://doi.org/10.1016/j.phytochem.2016.03.006 Get rights and content

Highlights

•
A polyoxypregnane aglycone and four polyoxypregnane steroids were isolated from Marsdenia tenacissima.
•
The polyoxypregnane steroids are naturally occurring polyoxypregnane glycosides bearing an open-chain sugar moiety.
•
Two of these compounds exhibited a wide spectrum of chemoresistance reversal activity.
•
Potential mechanisms were studied.

Abstract

A polyoxypregnane aglycone, 12β-O-acetyl-11α-O-isobutyryltenacigenin B, and four polyoxypregnane glycosides with a pachybionic acid ester moiety, 12β-O-acetyl-3-O-(6-deoxy-3-O-methyl-β-d-allopyranosyl-(1→4)-β-d-oleandronyl)-11α-O-isobutyryltenacigenin B, 12β-O-acetyl-3-O-(6-deoxy-3-O-methyl-β-d-allopyranosyl-(1→4)-β-d-oleandronyl)-11α-O-tigloyltenacigenin B, 12β-O-acetyl-3-O-(6-deoxy-3-O-methyl-β-d-allopyranosyl-(1→4)-β-d-oleandronyl)-11α-O-2-methylbutyryltenacigenin B, and 12β-O-acetyl-3-O-(β-d-glucopyranosyl-(1→4)-6-deoxy-3-O-methyl-β-d-allopyranosyl-(1→4)-d-oleandronyl)-11α-O-tigloyltenacigenin B, were isolated from the canes of Marsdenia tenacissima, together with a disaccharide derivative. Their structures were elucidated by extensive spectroscopic analysis, and the absolute configurations were further determined by X-ray crystallographic analysis. With the exception of the disaccharide derivative, all five compounds are unusual naturally occurring polyoxypregnane glycosides bearing an open-chain sugar moiety. Two of these exhibit a wide spectrum of chemoresistance reversal activity, and potential mechanisms were studied accordingly.

Graphical abstract

A polyoxypregnane aglycone (1) and four polyoxypregnane steroids (2–5) possessing an open-chain sugar moiety were isolated from Marsdenia tenacissima. Compounds 3 and 4 exhibit a wide spectrum of chemoresistance reversal activity, and potential mechanisms were studied accordingly.

Introduction

Marsdenia tenacissima (Roxb.) Wight et Arn., a plant belonging to the family Asclepiadaceae, is widely distributed from tropical to subtropical Asia. Its rhizomes and roots have long been used in China for the treatment of asthma, cancer, tracheitis, tonsillitis, pharyngitis, cystitis, and pneumonia (Jiangsu New College of Medicine, 1977). Previous chemical investigations resulted in the identification of more than 15 polyoxypregnane genins from M. tenacissima, classified into three types: tenacigenins A, B, and C (Yang et al., 1981) and their analogues (Deng et al., 2005a); 17β-tenacigenin B (Deng et al., 2005b) and the diester derivatives of tenacigenin B (Luo et al., 1993); and marstenacigenins A and B (Qiu et al., 1996). On the basis of the above aglycones, over 20 glycosides of tenacigenin B have been reported: from M. tenacissima tenacissosides A–N (Chen et al., 1999, Liu et al., 2008, Miyakawa et al., 1986, Wang et al., 2006, Xing et al., 2004) and marsdenosides A–K (Deng et al., 2005a, Deng et al., 2005c); seven glycosides of 17β-tenacigenin B named marsdenosides L and M (Huang et al., 2009) and tenacissimosides E–I (Yao et al., 2014), as well as four marstenacigenin glycosides named marstenacissides A–D (Xia et al., 2011). In addition, cissogenin (another type of compound) and its analogues, have been reported from seeds of the plant growing in India (Singhal et al., 1980a, Singhal et al., 1980b). Biological evaluations of extracts and isolated compounds from M. tenacissima also have antiasthmatic (Zhou et al., 1980), anticancer (Xue et al., 2012, Ye et al., 2014), and multidrug resistance (MDR) reversal (Han et al., 2012, Hu et al., 2008) activities. Furthermore, a new polyoxypregnane aglycone from this plant was able to circumvent P-glycoprotein (P-gp)-mediated MDR, as well as potentiating the activity of erlotinib and gefitinib in epidermal growth factor receptor tyrosine kinase inhibitor (EGFR TKI)-resistant non-small-cell lung cancer cells (Yao et al., 2014).

In a continuation of efforts in the search for MDR-reversal agents from natural sources, the EtOAc fraction of the EtOH extract prepared from the stems of M. tenacissima was investigated. A polyoxypregnane aglycone (1) and four new C₂₁ steroids with an open-chain sugar moiety (2–5), together with a known disaccharide derivative (6), were afforded. Here, the isolation and structural characterization of these new compounds, their chemoresistance reversal activity, and mechanistic studies are presented.

Section snippets

Structure elucidation

A 95% EtOH extract of the air-dried stems of M. tenacissima (8.0 kg) was suspended in water and extracted with EtOAc. The EtOAc fraction was dissolved in MeOH, and partitioned into a MeOH and a petroleum ether extract. After repeated column chromatography over silica gel, Sephadex LH-20, and MCI gel, and preparative HPLC, compounds 1–6 were obtained (Fig. 1).

Compound 1 was obtained as a colorless, amorphous powder. Its HR-ESI-MS peak at m/z 499.2642 ([M+Na]⁺, calculated for C₂₇H₄₀O₇Na 499.2672)

Conclusions

Compounds 3–5 represent the first examples of naturally occurring polyoxypregnane glycosides bearing an open-chain sugar moiety. It is assumed that there might be two possible pathways for the formation of this type of compound. Taking compound 3 as an example (Scheme 3), its formation may involve enzymatic oxidation of the first anomeric carbon of its precursor (see path I), or probably is associated with nucleophilic attack of compound 6 by 1a, which leads to lactone cleavage and generation

General experimental procedures

Optical rotations were measured on a Perkin–Elmer M341 polarimeter, whereas IR spectra were obtained on a Perkin–Elmer 577 spectrophotometer using KBr disks. NMR spectra were recorded on Varian Mercury-plus 300, Varian Mercury-plus 400, Bruker Avance III 400, Bruker Avance III 500, and Varian Inova 600 NMR spectrometers. Chemical shift (δ) values are given in ppm with TMS as internal standard, and coupling constants (J) are in Hz. LR-ESI-MS were recorded on a Waters 3100 Mass Detector and

Acknowledgements

The authors thank Professor Xuefeng Mei from the Shanghai Institute of Materia Medica for the X-ray diffraction experiment. Financial support from National Science & Technology Major Project “Key New Drug Creation and Manufacturing Program” (No. 2012ZX09301001-001), National Natural Science Funds of China (No. 81302657), Ministry of Science and Technology (2010DFA30980), Chinese Academy of Sciences (KSZD-EW-Z-004-01), Youth Innovation Promotion Association CAS, Shanghai Commission of Science

References (37)

J. Deng et al.
Marsdenosides A–H, polyoxypregnane glycosides from Marsdenia tenacissima
Phytochemistry
(2005)
J.Q. Feng et al.
Immunosuppressive pregnane glycosides from Periploca sepium and Periploca forrestii
Phytochemistry
(2008)
S.Y. Han et al.
Marsdenia tenacissima extract restored gefitinib sensitivity in resistant non-small cell lung cancer cells
Lung Cancer
(2012)
J. Liu et al.
A new C₂₁ steroid glycoside from Marsdenia tenacissima
Chin. Chem. Lett.
(2008)
S. Miyakawa et al.
Five glycosides from the Chinese drug “tong-guang-san”: the stems of Marsdenia tenacissima
Phytochemistry
(1986)
S. Singhal et al.
Cissogenin, a pregnane genin from Marsdenia tenacissima
Phytochemistry
(1980)
S. Singhal et al.
Tenasogenin, a pregnane ester from Marsdenia tenacissima
Phytochemistry
(1980)
K.K. To et al.
Protein phosphatase 2A inhibition and circumvention of cisplatin cross-resistance by novel TCM-platinum anticancer agents containing demethylcantharidin
Bioorg. Med. Chem.
(2004)
E. Abisch et al.
Die Glykoside der Wurzeln von Pachycarpus lineolatus (DECNE.) BULLOCK (oder P. schweinfurthii (N. E. BR.) BULLOCK) Glykoside und Aglykone, 201. Mitteilung
Helv. Chim. Acta
(1959)
J.J. Chen et al.
New C₂₁ steroidal glycosides from Marsdenia tenacissima
Acta Bot. Yunnanica
(1999)

J. Deng et al.

Two new C21 steroids from Marsdenia tenacissima

Chin. Chem. Lett.

(2005)

J. Deng et al.

Three new polyoxypregnane glycosides from Marsdenia tenacissima

Helv. Chim. Acta

(2005)

Y.J. Hu et al.

Tenacigenin B derivatives reverse P-glycoprotein-mediated multidrug resistance in HepG2/Dox cells

J. Nat. Prod.

(2008)

X.D. Huang et al.

Two new polyoxypregnane glycosides from Marsdenia tenacissima

Helv. Chim. Acta

(2009)

K.A. Jaeggi et al.

Die Glykoside der Wurzeln von Gongronema taylorii (SCHLTR. & RENDLE) BULLOCK. Glykoside und Aglykone, 292. Mitteilung

Helv. Chim. Acta

(1967)

Jiangsu New College of Medicine

A Dictionary of Traditional Chinese Drugs

(1977)

S. Kawanishi et al.

Constituents of Chinese crude drug “Wujiapi.” VVIII. On the structures of new oligosaccharides C1, D2, F1 and F2 of Bei-Wujiapi

Chem. Pharm. Bull.

(1977)

G.M. Lai et al.

P-glycoprotein expression and schedule dependence of adriamycin cytotoxicity in human colon carcinoma cell lines

Int. J. Cancer

(1991)

Cited by (19)

Pregnane steroids and steroid glycosides with multidrug resistance reversal activity from Marsdenia tenacissima
2023, Fitoterapia
Twenty compounds comprising four pregnane steroids (2–4 & 20) and 16 pregnane glycosides (1 & 5–19) have been obtained from the ethanol extract of the roots of a Dai ethnological herb, Marsdenia tenacissima. Their structures were characterized on the basis of comprehensive spectroscopic analyses with 17 ones (1–17) being reported for the first time, including the rare cases (2 & 3) of free C₂₁ steroids with 17α-acetyl substitution, compounds 4–7 bearing an unusual 14α-OH, and the first examples with simultaneous 14α-OH/17α-acetyl substitution (7) and glycosylation at C-12 position (10 & 11). An empirical rule for the identification of C-17 configuration, in C₂₁ steroids incorporating the marsdenin constitution structure, was also proposed. All the isolates, along with an array of previously reported analogues in our compound library, were screened for their chemo-reversal ability against P-glycoprotein (P-gp)-mediated multidrug resistance (MDR) in MCF-7/ADR cell line, and six compounds exhibited moderate MDR reversal activity with reversal folds ranging from 1.92 to 4.44.
Marsdenia tenacissima injection induces the apoptosis of prostate cancer by regulating the AKT/GSK3β/STAT3 signaling axis
2023, Chinese Journal of Natural Medicines
Marsdenia tenacissima injection, a standard Marsdenia tenacissima extract (MTE), has been approved as an adjuvant therapeutic agent for various cancers. Our previous study showed that MTE inhibited the proliferation and metastasis of prostate cancer (PCa) cells. However, the underlying mechanisms and active ingredients of MTE against PCa were not completely understood. This study revealed that MTE induced significant decreases in cell viability and clonal growth in PCa cells. In addition, MTE induced the apoptosis of DU145 cells by reducing the mitochondrial membrane potential and increasing the expression of Cleaved Caspase 3/7, Cyt c, and Bax. In vivo, DU145 xenografted NOD-SCID mice treated with MTE showed significantly decreased tumor size. TUNEL staining and Western blot confirmed the pro-apoptotic effects of MTE. Network pharmacology analysis collected 196 ingredients of MTE linked to 655 potential targets, and 709 PCa-associated targets were retrieved, from which 149 overlapped targets were screened out. Pathway enrichment analysis showed that the HIF-1, PI3K-AKT, and ErbB signaling pathways were closely related to tumor apoptosis. Western blot results confirmed that MTE increased the expression of p-AKT^Ser473 and p-GSK3β^Ser9, and decreased the expression of p-STAT3^Tyr705in vitro and in vivo. A total of 13 compounds in MTE were identified by HPLC-CAD-QTOF-MS/MS and UPLC-QTOF-MS/MS. Molecular docking analysis indicated that six compounds may interact with AKT, GSK3β, and STAT3. In conclusion, MTE induces the endogenous mitochondrial apoptosis of PCa by regulating the AKT/GSK3β/STAT3 signaling axis, resulting in inhibition of PCa growth in vitro and in vivo.
Tongguanteng injection reverses paclitaxel resistance via upregulation of TAB1 expression in ovarian cancer in vitro and in vivo
2023, Journal of Ethnopharmacology
Tongguanteng injection (TGT), the water extract from the stem of the Traditional Chinese hebal medicine of Marsdenia tenacissima (Roxb.) Wight et Arn. has been used as anticancer remedy for decades. TGT was not only used in the treatment of many malignant cancers extensively, but also an adjuvant anticancer drug with chemotherapeutics clinically.
To evaluate the effects of TGT on reversing paclitaxel (PTX) resistance and investigate the potential mechanism related to TAB1 in ovarian cancer (OC) in vitro and in vivo.
The synergistic effect and reversal ratio were determined by CCK8 assay and median-effect principle after the combination of TGT and PTX in OC A2780 and its PTX-resistant (A2780/T) cells. The biological functions in cell apoptosis, migration and invasion of A2780/T cells treated by PTX 4 μM with TGT 20, 40, 80 mg⋅mL^-1 for 24 h were evaluated by colony formation, flow cytometry, wound healing and transwell assays. Proteomics technique and bioinformatic analysis were used to indentify the change of TAB1 expression in A2780/T cells induced by TGT. The association between TAB1 expression and human OC was analyzed by gene expression databases. In A2780/T cells, western blotting and colony formation assays were used to investigate the relationship between TAB1 expression and PTX resistance after TAB1 overexpression by TAB1 plasmids. The mechanism of TGT and PTX regulating TAB1 and its related proteins were explored by western blotting and flow cytometry assays after TAB1 knock-down using siTAB1. Moreover, TUNEL staining, immunohistochemistry (IHC) and histopathology were used to observe the antitumor effects, TAB1 and p-p38 expression and the tissues impairments in nude mice xenograft model established by A2780/T cells after the co-treatment with TGT and PTX by in vivo.
TGT combined with PTX showed the synergistic effect (CI<1), which could reverse the IC₅₀ values of PTX in OC A2780 and A2780/T cells about 23.50 and 6.44 times, respectively. Besides, TGT combined with PTX could significantly inhibit the migration, invasion and promote apoptosis of A2780/T cells. We identified that TGT could induce TAB1 expression in A2780/T cells by proteomics analysis. TAB1 downregulation was significantly associated with tumorigenesis and poor prognosis in OC patients and PTX resistance in A2780/T cells. Furthermore, TGT could activate TAB1/TAK1/p38 MAPK signaling pathway targeting TAB1 and regulate the expression of Bax, Bcl-2 proteins to improve the sensitivity of A2780/T cells to PTX. TGT combined with PTX also showed a greater inhibition in tumor growth than PTX monotherapy in vivo. These promising results show the efficacy of TGT in reversing PTX resistance and provide a potential strategy that targeting TAB1/TAK1/p38 MAPK signaling pathway may improve the chemotherapy sensitivity in OC.
Our results revealed that Tongguanteng injection could reverse paclitaxel resistance and the potential mechanism might be associated with the activation of TAB1/TAK1/p38 MAPK signaling pathway in OC in vitro and in vivo. TAB1 might be a pivotal target for reversing PTX resistance. This study will provide a theoretical basis for the combination of Tongguanteng injection and paclitaxel in clinic.
Study on the molecular structure and chemical properties of the polyoxypregnane derivatives 11α-O-2-methylbutyryl-12β-O-tigloyl tenacigenin B and 11α,12β-O-ditigloyl tenacigenin B by combining experimental and theoretical methods
2021, Journal of Molecular Structure
Citation Excerpt :
Wight et Arn (Asclepiadaceae) is mainly distributed in the southwest of mainland China, whose dried rhizomes and roots have long been used either alone or in combination with other chemotherapeutics for the treatment of cancer [1]. In this plant, polyoxypregnane derivatives, including 11α-O-2-Methylbutyryl-12β-O-tigloyl tenacigenin B (11αM12βTTB) and 11α,12β-O-ditigloyl tenacigenin B (11αT12βTTB), were found to be the main ingredients and they have a wide spectrum of pharmaceutical properties, such as anti-inflammatory [2], anti-HIV [3], cytotoxic activities [4–6] and remarkable multidrug resistance reversal effects [7–11]. Both 11αM12βTTB and 11αT12βTTB, belong to the diester derivatives of tenacigenin B, the former with a 2-methylbutyryl group at sites C-11 while the latter with a tigloyl groups at sites C-11.
The title compounds, 11α-O-2-methylbutyryl-12β-O-tigloyl tenacigenin B and 11α,12β-O-ditigloyl tenacigenin B were isolated from the rhizomes and roots of Marsdenia tenacissima. The molecular geometries were optimized using DFT approach applying B3LYP/6–311G(d,p) base set and confirmed by infrared, nuclear magnetic resonance (NMR), Raman and UV–visible spectroscopic methods (experimental and theoretical). The present experimental data were found in reasonable agreement with the computational results. The vibrational features and the potential energy distribution (PED) of the title compounds were predicted. Moreover, the HOMOs and LUMOs analyses, total (TDOS) and (PDOS) partial density of states, chemical reactivity parameters, molecular electrostatic potential (MEP) and the intramolecular interactions such as topological analysis, natural bond orbital (NBO), reduced density gradient (RDG) of the optimized geometries were also performed. Nonlinear optical properties (NLO) for the title compounds were predicted. Besides, the molecular docking was investigated with the protein-PDB:3iw4 to check the mode of interactions. Therefore, these studies give important information pertaining to the true geometric configuration, stability, reactivity and bioactivity of the title compounds and show that 11αM12βTTB and 11αT12βTTB could have a great effect in the anticancer activity due to its protein active sites affinity and binding energy. The results are beneficial for the further research in order to design them as the novel drugs.
Polyoxypregnanes as safe, potent, and specific ABCB1-inhibitory pro-drugs to overcome multidrug resistance in cancer chemotherapy in vitro and in vivo
2021, Acta Pharmaceutica Sinica B
Multidrug resistance (MDR) mediated by ATP binding cassette subfamily B member 1 (ABCB1) is significantly hindering effective cancer chemotherapy. However, currently, no ABCB1-inhibitory drugs have been approved to treat MDR cancer clinically, mainly due to the inhibitor specificity, toxicity, and drug interactions. Here, we reported that three polyoxypregnanes (POPs) as the most abundant constituents of Marsdenia tenacissima (M. tenacissima) were novel ABCB1-modulatory pro-drugs, which underwent intestinal microbiota-mediated biotransformation in vivo to generate active metabolites. The metabolites at non-toxic concentrations restored chemosensitivity in ABCB1-overexpressing cancer cells via inhibiting ABCB1 efflux activity without changing ABCB1 protein expression, which were further identified as specific non-competitive inhibitors of ABCB1 showing multiple binding sites within ABCB1 drug cavity. These POPs did not exhibit ABCB1/drug metabolizing enzymes interplay, and their repeated administration generated predictable pharmacokinetic interaction with paclitaxel without obvious toxicity in vivo. We further showed that these POPs enhanced the accumulation of paclitaxel in tumors and overcame ABCB1-mediated chemoresistance. The results suggested that these POPs had the potential to be developed as safe, potent, and specific pro-drugs to reverse ABCB1-mediated MDR. Our work also provided scientific evidence for the use of M. tenacissima in combinational chemotherapy.
Sesquiterpenoids and triterpenoids from Secamone lanceolata blume with inhibitory effects on nitric oxide production
2019, Fitoterapia
Citation Excerpt :
S. lanceolata is a kind of high-quality fodder for pigs and sheep [1], and its leaves and flowers have been used in traditional Chinese medicine for the treatment of detumescence and eliminating stagnation, especially scrofula [2,3]. Previous phytochemistry studies revealed that Asclepiadaceae family were rich in steroids [4–7], cardiac glycosides [8,9], and alkaloids [10]. However, the reports about sesquiterpenoids and triterpenoids were quite limited.
Five previously undescribed sequiterpenoids, including three guaiane sesquiterpenoids secamonol A (2), secamonester (3), secamonether (4), and one elemanolide sequiterpenoid secamonol B (7), and one eudesmane sesquiterpenoid secamonol C (8) along with nine known compounds were isolated from Secamone lanceolata Blume. Their structures and absolute configurations were established by analyzing of NMR, CD, MS spectrum and single-crystal X-ray diffraction, as well as comparing with the literatures. Compounds 1 and 4–8 exhibited potential inhibition activities of NO production with IC₅₀ values in the range of 27.9–57.5 μM with no obvious cytotoxicities even at the high concentration (100 μM). Whereas, compound 3 showed significant cytotoxicity with CC₅₀ value of 5.6 μM against macrophages RAW264.7 Cells.

View all citing articles on Scopus

¹: Contributed equally to this work.

View full text

Polyoxypregnane steroids with an open-chain sugar moiety from Marsdenia tenacissima and their chemoresistance reversal activity

Highlights

Abstract

Graphical abstract

Introduction

Section snippets

Structure elucidation

Conclusions

General experimental procedures

Acknowledgements

Phytochemistry

Phytochemistry

Lung Cancer

Chin. Chem. Lett.

Phytochemistry

Phytochemistry

Phytochemistry

Bioorg. Med. Chem.

Die Glykoside der Wurzeln von Pachycarpus lineolatus (DECNE.) BULLOCK (oder P. schweinfurthii (N. E. BR.) BULLOCK) Glykoside und Aglykone, 201. Mitteilung

Helv. Chim. Acta

New C21 steroidal glycosides from Marsdenia tenacissima

Acta Bot. Yunnanica

Two new C21 steroids from Marsdenia tenacissima

Chin. Chem. Lett.

Three new polyoxypregnane glycosides from Marsdenia tenacissima

Helv. Chim. Acta

Tenacigenin B derivatives reverse P-glycoprotein-mediated multidrug resistance in HepG2/Dox cells

J. Nat. Prod.

Two new polyoxypregnane glycosides from Marsdenia tenacissima

Helv. Chim. Acta

Die Glykoside der Wurzeln von Gongronema taylorii (SCHLTR. & RENDLE) BULLOCK. Glykoside und Aglykone, 292. Mitteilung

Helv. Chim. Acta

A Dictionary of Traditional Chinese Drugs

Constituents of Chinese crude drug “Wujiapi.” VVIII. On the structures of new oligosaccharides C1, D2, F1 and F2 of Bei-Wujiapi

Chem. Pharm. Bull.

P-glycoprotein expression and schedule dependence of adriamycin cytotoxicity in human colon carcinoma cell lines

Int. J. Cancer

New C₂₁ steroidal glycosides from Marsdenia tenacissima