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To solve the problem of inaccurate prediction caused by the lack of representativeness of samples due to the small sample size of the collected clinical data when using machine learning methods to predict drug concentration in plasma and describe the hysteresis phenomenon of drug effect lagging behind plasma drug concentration, this paper proposes a pharmacokinetic-pharmacodynamic (PK-PD) model based on the SSA-1DCNN-Attention network and the semicompartment method. First, a one-dimensional convolutional neural network (1DCNN) is established, and the attention mechanism is introduced to determine the importance of each physiological and biochemical parameter. The sparrow search algorithm (SSA) is used to optimize the parameters of the network to improve the prediction accuracy after data enhancement through the synthetic minority oversampling technique (SMOTE) method. After constructing the time-concentration relationship of the drug through the SSA-1DCNN-Attention network, the concentration-effect relationship of the drug is established by using the semicompartment method to synchronize the drug effect with the concentration. At last, the phenobarbital (PHB) combined with Cynanchum otophyllum saponins to treat epilepsy was taken as an example to validate the proposed method.

From the Cynanchum wilfordii roots, 32 compounds, including 5 previously undescribed (1, 4-6, 12) and 27 known (2, 3, 7-11, 13-32) compounds, were isolated, and their structures were elucidated using NMR spectroscopic data and MS data aided by ECD calculations or the modified Mosher's reaction. All isolates were tested for their inhibitory effects on proprotein convertase subtilisin/kexin type 9 (PCSK9) secretion. Among the isolates, compound 4, a methyl cholesterol analog, exhibited the most potent effect in reducing PCSK9 secretion, along with PCSK9 downregulation at the mRNA and protein levels via FOXO1/3 upregulation. Moreover, compound 4 attenuated statin-induced PCSK9 expression and enhanced the uptake of DiI-LDL low-density lipoprotein. Thus, compound 4 is suggested to be a potential candidate for controlling cholesterol levels., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Chemical investigation of the acid hydrolysate of Cynanchum bungei roots led to the isolation of eleven undescribed steroids, namely cynbungenins A-K (1-11), and seven previously described analogues (12-18). The complete structures of these compounds were elucidated using the comprehensive spectroscopic analyses and reference data. Structurally, compounds 1 and 2 represent the first example of androstane-type steroids found in the Cynanchum plants, and compounds 3-6 and 12 are characterized as pregnane-type steroids with a rare 8,14-seco-steroid core. In the cytotoxic activity assay, compound 16 displayed the strongest cytotoxic effect against MCF-7, HCT-116, HeLa, and HepG2 cancer cell lines, with IC 50 values of 9.98-16.42 μM, and further research indicated that it induced both apoptosis and cell cycle arrest in the G0/G1 phase in a dose-dependent manner toward HepG2 cells., Competing Interests: Declaration of competing interest All authors declare that No conflict of interest exists., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Objective: To explore the targets and pathways of Cynanchum wilfordii for treatment of ulcerative colitis (UC)., Methods: UPLC-QE-MS was used to identify the components of Cynanchum wilfordii ethanol extract, and their targets were screened using public databases for construction of the core protein-protein interaction (PPI) network and GO and KEGG enrichment analyses. Forty male C57 mice were randomized into normal control group, model group, mesalazine group and Cynanchum wilfordii group ( n =10), and in the latter 3 groups, mouse UC models were established by treatment with 2.5% DSS and the latter 2 groups drug interventions by gavage. The therapeutic effect was evaluated by recording body weight changes and DAI score. Pathological changes of the colon tissue were observed with HE and AB-PAS staining, and JAK2 and STAT3 protein expressions were detected with Western blotting. The metabolites and metabolic pathways were identified by metabonomics analysis., Results: We identified 240 chemical components in Cynanchum wilfordii alcoholic extracts, including 19 steroids. A total of 177 Cynanchum wilfordii targets, 5406 UC genes, and 117 intersection genes were obtained. JAK2 and STAT3 were the core targets and significantly enriched in lipid and atherosclerosis pathways. Cynanchum wilfordii treatment significantly increased the body weight and decreased DAI score of UC mice ( P < 0.05), alleviated intestinal pathologies, and decreased JAK2 and STAT3 protein expressions in the colon tissues. Most of the 83 intersecting differential metabolites between the control, model and Cynanchum wilfordii groups were identified as glycerophospholipids, arachidonic acid, and amino acids involving glycerophospholipid metabolism and other pathways. Correlation analysis suggested that the core targets of Cynanchum wilfordii for UC participated in regulation of the metabolites., Conclusion: Cynanchum wilfordii alleviates lipid and amino acid metabolism disorders to lessen UC in mice by regulating the core targets including JAK2 and STAT3 and the levels of endogenous metabolites.

Two new pregnane glycosides ( 1 and 2 ), together with four known ones ( 3 - 6 ), were isolated from the roots of Cynanchum auriculatum Royle ex Wight (Asclepiadaceae). On the basis of detailed spectroscopic analysis and chemical method, the structures of new compounds were characterized to be metaplexigenin 3- O - β -D-cymaropyranosyl- (1→4)- α -L-diginopyranosyl-(1→4)- β -D-cymaropyranoside ( 1 ), metaplexigenin 3- O - α -L-diginopyranosyl-(1→4)- β -D-cymaropyranoside ( 2 ). All the isolated compounds ( 1 - 6 ) were tested for their in vitro inhibitory activity against the growth of human colon cancer cell lines HCT-116. Compounds 5 and 6 showed significant cytoxicities with IC 50 values of 43.58 µM and 52.21 µM.

Cynanotophyllosides E-F, two new minor pregnane glycosides were isolated from the antidepressant active fraction of cultivated Cynanchum otophyllum , and their structures were determined as 12- O -vanilloyl-deacetylmetaplexigenin 3- O - β -D-glucopyranosyl-(1→4)- β -D-glucopyranosyl-(1→4)- β -D-cymaropyranosyl-(1→4)- β -D-oleandropyranosyl-(1→4)- β -D-digitoxopyranoside, and 12- O -nicotinoyl-deacetylmetaplexigenin 3- O - β -D-glucopyranosyl-(1→4)- β -D-glucopyranosyl-(1→4)- β -D-cymaropyranosyl-(1→4)- β -D-oleandropyranosyl-(1→4)- β -D-cymaropyranoside respectively, with the combination of spectroscopic and chemical analysis.

Cynasibirolide A (1), one new humulanolide sesquiterpene, together with four known analogs, asteriscanolide (2), (1S,8S)-8-hydroxyhumula-2Z,6E,9E-trien-1,12-olide (3), (1S,7R)-8-oxohumula-2Z,9E-dien-1,12-olide (4), and (+)-6,7,9,10-tetrahydroasteriscunolide (5) were isolated from the roots and rhizomes of Cynanchum acutum subsp. sibiricum. Their structures and configurations were elucidated by spectroscopic methods, including 2D-NMR techniques, and the structure of 1 was confirmed by single-crystal X-ray diffraction. All compounds were evaluated for their anti-complementary activity in vitro, and compound 3 exhibited anti-complement effect with CH 50 value of 0.45 mM., (© 2023 Wiley-VHCA AG, Zurich, Switzerland.)

Five new C 21 -steroidal sapogenins (1-5) named cynotogenins J-N, were isolated from the acid hydrolysate of Cynanchum otophyllum roots. Their structures were established by extensive spectroscopic analysis (UV, IR, HR-ESI-MS, and NMR). Most notably, compounds 1-3 harboring a rare 5β,6β-epoxy group in the C 21 -steroidal skeleton of Cynanchum plants. All compounds were evaluated for their cytotoxicities against multiple cancer cell lines, in which compounds 5 showed weak cytotoxicity against HepG2 cancer cells with IC 50 values of 44.90 μM., (© 2023 Wiley-VHCA AG, Zurich, Switzerland.)

Tobacco mosaic virus (TMV) is known to infect a wide range of plants, resulting in reduced yield and productivity. Novel, effective, and plant-based pesticides are required to protect plants against TMV infection. To identify novel anti-TMV agents from natural sources, we systematically studied the roots of Cynanchum paniculatum and isolated six new seco-pregnane C 21 steroidal glycosides, along with 14 known compounds. Their structures were elucidated by comprehensive spectroscopic data analysis. The anti-TMV activity of compounds were screened using the half-leaf method. Biological tests revealed that compounds 1, 2, 5, 9, 10, 15, and 16 displayed significant anti-TMV activities compared with the positive control ningnanmycin. In addition, reverse transcription-polymerase chain reaction (RT-PCR) and western blot analysis confirmed the antiviral activity of these compounds, as evident from reduced TMV coat protein (TMV-CP) gene replication and TMV-CP protein expression. These compounds downregulated the expression of NtHsp70-1 and NtHsp70-261, indicating that these steroidal glycosides possibly inhibit the TMV infection by suppressing the expression of NtHsp70-1 and NtHsp70-061 expression., (Copyright © 2022. Published by Elsevier B.V.)

For our interest in the potential biologically active and structurally unique steroidal glycosides, continued phytochemical investigation of Cynanchum taihangense was carried out; twelve new seco-pregnane glycosides, cynataihosides I-L ( 1 - 4 ), M-T ( 7 - 14 ), and two known glycosides, glaucoside A ( 5 ) and atratcynoside F ( 6 ), were isolated from the 95% ethanol extract of Cynanchum taihangense . Two new aglycones were found among compounds 10 , 11 , 13 , and 14 . The structures of the glycosides were elucidated based on 1D and 2D NMR spectroscopic data, HR-ESI-MS analysis, and chemical evidence. The cytotoxicity of compounds against three human tumor cell lines (HL - 60, THP-1, and PC-3) were evaluated by MTT assay. Compound 11 displayed significant cytotoxicity against THP-1 and PC-3 cell line with IC 50 values of 5.08 and 22.75 μm, respectively. Compounds 3 and 14 exhibited moderate and selective cytotoxicity on HL - 60 and THP-1 with IC 50 values of 17.78 and 16.02 μm, respectively.

Background: Bungarus multicinctus is one of the top ten venomous snakes in China. Its venom is mainly neurotoxin-based. Novel antivenom drugs need to be further researched and developed., Objective: This study aimed to explore the molecular mechanism of Cynanchum paniculatum in treating Bungarus multicinctus bites based on network pharmacology. Material and methods . The potential active ingredients of Cynanchum paniculatum were screened and their SDF structures were obtained using the PubChem database and imported into the SwissTargetPrediction database, and targets were obtained for the antitoxin effects of Cynanchum paniculatum in the treatment of Bungarus multicinctus bites. The Cynanchum paniculatum-active compound-potential target network and protein-protein interaction network were constructed by using Cytoscape software, and then biological function analysis and KEGG pathway enrichment analysis were performed using the DAVID., Results: Seven potential active components (cynapanoside C, cynatratoside B, tomentolide A, sitosterol, sarcostin, tomentogenin, and paeonol) and 286 drug targets were obtained, including 30 key targets for the treatment of bungarotoxin toxicity. The active components mainly acted on PIK3CA, MAPK1, MAP2K1, JAK2, FYN, ACHE, CHRNA7, CHRNA4, and CHRNB2, and they antagonized the inhibitory effect of bungarotoxin on the nervous system through cholinergic synapses and the neurotrophin signaling pathway., Conclusions: Cynanchum paniculatum exerts a therapeutic effect on Bungarus multicinctus bites through multiple active components, multiple targets, and multiple pathways. The findings provide a theoretical basis for the extraction of active components of Cynanchum paniculatum and for related antivenom experiments., Competing Interests: The authors declare no competing interests., (Copyright © 2022 Linsheng Zeng et al.)

Six unidentified C 21 steroidal glycosides, cynwallosides A-F, as well as twenty-two known compounds, were isolated from the roots of Cynanchum wallichii Wight. The structures of cynwallosides A-F were determined by spectroscopic analysis and acidic hydrolysis. Most of these twenty-eight compounds were found to significantly reverse drug resistance in both the MCF-7/ADR and HepG2/ADM cell lines by suppressing P-gp protein expression. Further investigation revealed that three compounds suppressed P-gp expression by significantly inactivating the JNK and NF-κB pathways., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The study investigated the difference of intestinal absorption characteristics of root tuber of Cynanchum auriculatum extract between normal and functional dyspepsia(FD) model rats with everted intestine sac model.The content of syringic acid, scopoletin, caudatin, baishouwu benzophenone, qingyangshengenin and deacyhmetaplexigenin in the C.auriculatum extract in different intestinal segments was detected by UPLC-MS/MS.The cumulative absorption amount(Q) and absorption rate constant(K&#95;a) of the six chemical constituents were calculated.The results showed that the six components could be absorbed into the intestinal sac and were unsaturated, which indicated that the absorption mechanism of scopoletin was active transport in the intestine, while that of the other five components were passive diffusion.For normal group, the syringic acid and baishouwu benzophenone in ileum, qingyangshengenin and deacyhmetaplexigenin in ileum and duodenum, and caudatin in colon were well absorbed and scopoletin at low, medium and high concentrations was found excellent absorption in jejunum, ileum, and colon, respectively.Whereas the best absorption site of each component was ileum in model group.The absorption characteristics of each component between normal group and model group were complex at different concentrations, showing inconsistent tendency of absorption, which suggested that the components of root tuber of C.auriculatum extract were selectively absorbed in small intestine, and the absorption characteristics of the six components could be changed under FD status.This study provided theoretical basis for the clinical drug application and development of root tuber of C.auriculatum.

Three new compounds named cynansteroid A (1), cynansteroid B (2) and cynansteroid C (3), together with nine known C 21 -steroidal pregnane sapogenins (4-12) were isolated from the hydrolytic extract of the roots of Cynanchum auriculatum. The structures of cynansteroid A-C (1-3) were ascertained via the detailed analysis of the HR-ESI-MS, 1D and 2D NMR, and the calculated and experimental ECD data of cynansteroid B (2). Compound 11 displayed moderate inhibitory activity toward Verticillium dahliae Kleb (IC 50 =37.15 μM), furthermore, compounds 11 and 12 showed significant inhibitory activity against Phomopsis sp. (IC 50 =16.49 μM and 17.62 μM, respectively)., (© 2022 Wiley-VHCA AG, Zurich, Switzerland.)

One novel monoterpene rhamnoside (1) and 7 known monoterpenes (2-8) were isolated from the ethanol extract of Cynanchum atratum for the first time. Their structures were identified by comprehensive spectroscopic data analysis such as nuclear magnetic resonance, high-resolution electrospray ionization mass spectra, optical rotatory dispersion, and acid hydrolysis. In the subsequent antioxidant assay, compound 8 exhibited obvious 2,2-diphenyl-2-picrylhydrazyl hydrate radical scavenging activity., (© The Author(s) 2022. Published by Oxford University Press on behalf of Japan Society for Bioscience, Biotechnology, and Agrochemistry.)

Three new steroidal glycosides, metapregnoside A-C ( II - IV ), together with one known compound, byzantionoside B ( I ), were isolated from the fresh whole herb of Metaplexis japonica by using high-speed countercurrent chromatography and semi-preparative liquid chromatography. Their structures and relative configurations were elucidated by spectroscopic methods including 1D NMR, 2D NMR and HR-ESI-MS. The potential targets of compound I - IV were identified by virtual screening. And the potential inhibitory effects of these compounds on tyrosine protein kinases were compared by molecular docking. Byzantionoside B ( I ) was the first isolated compound from Metaplexis genus. The docking score of metapregnoside C ( IV ) was the highest. And the sugar chain residues at position C-20 in the pregn-4-en-3-one derivatives is the main factor affecting their docking scores on tyrosine protein kinases Fes/Fps.

Ethnopharmacological Relevance: Cynanchum komarovii Al. Iljinski is a ethnomedicinal herb and this ethno-medicine is used mainly to treat arthritis, toothache, reducing phlegm, relieving cough. Total alkaloids of Cynanchum komarovii Al. Iljinski (TACKI) is the main active compound of Cynanchum komarovii Al. Iljinski. Previous investigations have revealed that TACKI can significantly inhibit rat foot swelling caused by carrageenan; it has a significant inhibitory effect on granulation tissue proliferation. Pharmacology study showed that Cynanchum komarovii Al. Iljinski has analgesia, anti-inflammatory, antibacterial, anti-tumor, relieving cough and relieving asthma. However, there is no any investigation on the mechanism of analgesia and anti-inflammation., Aim of the Study: To clarify the analgesic effect and material basis of Cynanchum komarovii Al. Iljinski, determine the analgesic effect of TACKI, and provide experimental data support for its traditional application in the treatment of various pains., Materials and Methods: TACKI were prepared by the traditional acid extraction and alkaline precipitation method, and TACKI was analyzed through classic animal models of acute antinociceptive animal models and chronic antinociceptive. Evaluation of analgesic effects, and preliminary discussion of the mechanism of its analgesic effects were performed in this work., Results: Acute toxicity experiments showed that the LD 50 of TACKI mice was 2960.88 mg/kg, and symptoms of poisoning appeared. Patholog of liver and kidney studies have shown that TACKI reduces eosinophils and increases basophils in kidney glomeruli. In the study of analgesic effects, TACKI had analgesic activity through the PWL, formalin test, and acetic acid writhing test. In the chronic inflammatory antinociceptive study, the latency of the withdrawal reflex in the TACKI group was prolonged, and the mechanical withdrawal reflex threshold was significantly increased. The protein expression of NMDA, GFAP and Iba-1 in rat brain tissue can be reduced significantly byTACKI. Meanwhile, the content of TNF-α and IL-6 in rat brain tissue is reduced., Conclusion: TACKI has a significant analgesic activities. It may be related to inhibiting the activation of astrocytes and reducing the content of inflammatory mediators., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Cynanchum auriculatum Royle ex Wight. ( CA ), Cynanchum bungei Decne. ( CB ) and Cynanchum wilfordii (Maxim.) Hemsl. ( CW ) are three close species belonging to the Asclepiadaceous family, and their dry roots as the bioactive part have been revealed to exhibit anti-tumor, neuroprotection, organ protection, reducing liver lipid and blood lipid, immunomodulatory, anti-inflammatory, and other activities. Until 2021, phytochemistry investigations have uncovered 232 compounds isolated from three species, which could be classified into C 21 -steroids, acetophenones, terpenoids, and alkaloids. In this review, the morphology characteristics, species identification, and the relationship of botany, extraction, and the separation of chemical constituents, along with the molecular mechanism and pharmacokinetics of bioactive constituents of three species, are summarized for the first time, and their phytochemistry, pharmacology, and clinical safety are also updated. Moreover, the direction and limitation of current research on three species is also discussed.

Numerous C-21 steroidal glycosides have been isolated from Cynanchum plants. Many of them derive from the aglycone caudatin (CDT) which includes a tetracyclic deacylmetaplexigenin unit and an ikemaoyl ester side chain. CDT can be found in diverse traditional medicines, such as Baishouwu radix used to treat gastro-intestinal disorders. The compound has revealed marked anticancer properties, reviewed here. CDT and its mono-glycoside analogue CDMC display antiproliferative activities against different cancer cell lines in vitro and have revealed significant anticancer effects in tumor xenograft models in vivo. Their mechanism of action is multifactorial, implicating several signaling pathways (Wnt/GSK3/β-catenin, TRAIL/DR5/ER and TNFAIP1/NFκB) which contribute to the antiproliferative, antiangiogenic, antimetastatic and proapoptotic effects of the natural products. CDT also modulates DNA replication, is antioxidant and targets some cancer stem cells. CDT and CDMC are interesting anticancer products, while other CDT glycoside derivatives display antiviral and antifungal activities. Altogether, the present review provides a survey of the pharmacological profiles of CDT and derivatives. The lack of knowledge about the molecular targets of CDT currently limits drug development but the natural product, orally active, warrants further pharmacology and toxicology studies., (Copyright © 2021 Elsevier Inc. All rights reserved.)

As our ongoing chemical investigation, two new pregnane steroidal glycosides, cynataihosides G ( 1 ), with a new aglycone, and H ( 2 ) were isolated from the 95% ethanol extract of Cynanchum taihangense . Their structures were elucidated on the basis of 1 D and 2 D NMR spectral data, HR-ESI-MS analysis and qualitative chemical methods. The compounds were subjected to detect the cytotoxicity against three human tumor cell lines (HL-60, THP-1 and PC-3). The compounds displayed no significant cytotoxicity.Supplemental data for this article can be accessed at https://doi.org/10.1080/14786419.2019.1672682.

Neuroprotection under conditions of oxygen stress of two C 21 steroidal glycosides, cynsaccatol Q ( 1 ) and saccatol K ( 2 ), isolated from Cynanchum auriculatum Royle ex Wight, and their potential mechanisms were studied using the model of H 2 O 2 -induced damage on PC12 cells in this paper. Experiments showed that compounds 1 and 2 can both regulate the activities of antioxidant enzymes, including GSH-Px, LDH, CAT, SOD, decrease the levels of intracellular ROS and Ca 2+ , reduce cell apoptosis, and regulate the expression of HO-1 and NQO1 through the Nrf2-ARE pathway, thus play the role of neuroprotection against oxidative damage.

Naturally occurring C 21 -steroidal aglycones from Cynanchum exhibit significant antitumor effects. To expand the chemical diversity and get large scale C 21 -steroidal aglycones, the extracts of the roots of Cynanchum otophyllum were treated with 5% HCl in aqueous and the resulting hydrolysate was investigated. Nine new C 21 -steroidal aglycones (1-9) namely cynotogenins A-I, along with seventeen known analogous (10-26), were isolated from the hydrolysate. The structures of compounds 1-9 were elucidated by spectroscopic analysis (IR, HR-ESI-MS, 1D and 2D NMR) and comparison of observed spectroscopic data with those of reported in the literature. Aglycones 2-5 with rare cis-cinnamoyl group as well as 8 and 9 with 5β,6β-epoxy group were found from the genus of Cynanchum for the first time. The cytotoxicities of compounds 1-26 toward human cancer HeLa, H1299, HepG2, and MCF-7 cells were evaluated and preliminary structure-activity relationship (SAR) was discussed. Moreover, compound 20 inhibits HepG2 cell apoptosis and induces of G0/G1 phase arrest in a dose dependent manner., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Thrombosis causes poor blood circulation, which may lead to several cardiovascular disorders. Antiplatelet aggregation and antihyperlipidemia are the key processes that improve blood circulation. The antiplatelet aggregation and antihyperlipidemic effects of ACG-1, a mixture of Angelica gigas , Cynanchum wilfordii , and Ginkgo biloba extracts, were investigated in this study. The antiplatelet aggregation activity of ACG-1 was determined by studying its effects on collagen-induced platelet aggregation in human platelet-rich plasma (PRP). In addition, the effects of ACG-1 were investigated in a thromboembolism mouse model. The high-fat diet (HFD)-fed mouse model was used to investigate the antihyperlipidemic effects of ACG-1 and western blotting assay was performed to elucidate its mechanism of action. It was observed that ACG-1 significantly inhibited platelet aggregation in human PRP. Furthermore, ACG-1 showed protective effects in a thromboembolism mouse model induced by administering a mixed collagen and epinephrine intravenous injection. Oral administration of ACG-1 also significantly ameliorated blood lipid profiles in the HFD-fed mouse model. In conclusion, ACG-1 should be considered a powerful functional food to improve blood circulation.

The menopausal syndrome caused by rapid changes in hormone levels greatly influences the quality of life of women. Though hormone replacement therapy (HRT) is widely used to treat the menopausal syndrome, it exhibits many side effects, including the risk of thrombosis, cardiovascular diseases, and increased incidence of breast cancer; thus, diversifying the interest for phytotherapy-based materials as alternatives to HRT. Here, we isolated a crude polysaccharide fraction (CWPF) from Cynanchum wilfordii root that alleviated the ovariectomy-induced uterine atrophy and bone loss without changes in plasma estradiol concentration in mice. Increased plasma levels of follicle-stimulating hormone (FSH), alkaline phosphatase (ALP), osteocalcin (OC) in ovariectomized mice were also reduced to normal levels by CWPF administration. We found that the inhibitory effects of CWPF on menopausal symptoms were mediated by the estrogen receptor β (ER-β) specific activation, not ER-α. Moreover, CWPF treatment suppressed the phosphorylation of Akt, suggesting that CWPF alleviates post-menopausal symptoms by regulating ER-β related Akt signaling pathway. These results demonstrate that the polysaccharides corresponding to CWPF among the water-soluble extracts of CW could be used as a beneficial herbal alternative for the development of therapeutic agents to prevent menopausal syndrome in women., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

A chemical investigation of Cynanchum mongolicum (Maxim.) Kom. identified 8 compounds. On the basis of spectroscopic data, they were determined to be 3 alkaloids and 5 sinapoyl esters, among which were two previously undescribed compounds ( 1 and 2 ). The inhibitory effects of the isolated compounds against four human tumor cell lines were evaluated in vitro by MTT assays, which revealed moderate inhibitory effects with IC 50 values < 50 mM, in particularly, three antofine analogues have showed significant antitumor activities with IC 50 values < 0.1 mM, which was obviously better than the 5-fluorouracil and potential to be used as cancer drugs.

Ethnopharmacological Relevance: Cynanchum paniculatum (Bunge) Kitag. ex H. Hara (C. paniculatum), is a broadly used traditional medicinal plant by East Asians. The roots and rhizomes of this herb were named 'Xu-Chang-Qing' since the Qin or Han Dynasty (B.C.221-220) in China. It is pungent and warm in nature and associated with the liver and stomach meridians. Moreover, the efficacy of this herb are dispelling wind, resolving dampness, relieving pain and itching. It is used for treating the onset of rheumatic arthralgia, stomachache, toothache, lumbago, soft tissue injury, rubella and eczema., Aim of the Study: The purpose of this paper is to provide a systematic review about the botany, traditional uses, phytochemistry and pharmacology of C. paniculatum on the strength of the studies in the past two decades., Materials and Methods: A comprehensive search on previous literature was conducted on databases such as Web of Science, Pubmed, Sciencedirect, American Chemical Society (ACS), Google scholar and China national knowledge internet (CNKI). The search was based on the botany, traditional uses, phytochemistry and pharmacology of C. paniculatum. The key search words were 'Cynanchum paniculatum' and 'Radix Cynanchi Paniculati'. In addition, some published books were searched for more information on the herb., Results: Over 150 compounds have been isolated and identified from C. paniculatum, including C 21 steroids, volatile oils, carbohydrates and phenanthroindolizidine alkaloids. Extensive pharmacological activities of the extracts or compounds of C. paniculatum in vivo and in vitro were confirmed including anti-inflammatory, anti-nociceptive, sedative antiviral, antitumor, neuroprotective, treating snake bites, immunomodulatory, anti-radiation, vasodilatory, acaricidal potentials and anti-adipogenic activities., Conclusions: In this paper, the botany, traditional uses, phytochemistry and pharmacology of C. paniculatum were reviewed. This herb has long been used as traditional medicine. It was reported with numerous chemical ingredients and various pharmacological activities with anti-inflammatory, antitumor, neuroprotection, etc. In the future, C. paniculatum still needs further study, such as identifying the active compounds, clarifying the pharmacological mechanisms, discussing quality and safety., (Copyright © 2020. Published by Elsevier B.V.)

We herein show a dramatic change of herbal properties of the composition as well as function via fermentation of Cynanchi atrati Radix (family Asclepiadaceae). Cynanchi atrati Radix showed a high cytotoxicity against B16-F10 melanoma cell line, but the function of Cynanchi atrati Radix was completely changed into anti-melanin activity at very low concentration after Lactobacillus -fermentation. In addition, the compounds were drastically changed as shown in HPLC-based profile. Furthermore, this transformation has been achieved by only Lactobacillus -fermentation. This study proposes an strategy which we need to consider in the herb-derived material researches including pharmacopuncture., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2020. Published by Elsevier B.V.)

This trial aimed to determine the effect of a standardized Cynanchum wilfordii Radix extract (CWE) on the lipid profiles of individuals with elevated total cholesterol (T-Chol) using a double-blind randomized placebo-controlled design. Ninety-six Korean individuals with elevated T-Chol level (200-240 mg/dL) were recruited and randomly allocated to groups that received VasH300 (300 mg CWE/day, n = 32), VasH600 (600 mg CWE/day, n = 32), or a placebo (n = 32) groups. Primary outcomes included T-Chol, low-density lipoprotein (LDL)-cholesterol, high-density lipoprotein (HDL)-cholesterol, triglyceride, and safety (adverse events, biochemical parameters, and hematological parameters). Data were compared using a one-way analysis of variance followed by Duncan's post-hoc tests (among groups) and paired t tests (within groups). Values for T-Chol and LDL-cholesterol were significantly reduced in the VasH300 and groups (VasH300: 4.0 and 6.4%, respectively; VasH600; 3.8 and 5.8% respectively; both p < .05) compared with the placebo group and were not dose-dependent. VasH300 significantly improved the lipid profiles of individuals with elevated T-Chol without any serious side effects. Daily supplementation with VasH might be an alternative strategy with which to modify cholesterol-related parameters, especially in individuals with elevated T-Chol levels., (© 2020 John Wiley & Sons, Ltd.)

In the complex tumor microenvironment, cancer stem cells (CSCs), a rare population of cells, are responsible for malignant tumor initiation, metastasis, drug resistance and recurrence. Controlling breast CSCs (BCSCs) using natural compounds is a novel potential therapeutic strategy for clinical cancer treatment. In this study, a mammosphere assay-guided isolation protocol including silica gel, a C18 column, gel filtration, and high-pressure liquid chromatography was used to isolate an inhibitory compound from Cynanchum auriculatum extracts. The isolated inhibitory compound was identified as caudatin. Caudatin inhibited breast cancer cell proliferation, mammosphere formation and tumor growth. Caudatin decreased the CD44 + /CD24 - and aldehyde dehydrogenase + cell proportions and the levels of c-Myc, Oct4, Sox2, and CD44. Caudatin induced ubiquitin (Ub)-dependent glucocorticoid receptor (GR) degradation and blocked subsequent Yes-associated protein (YAP) nuclear accumulation and target gene transcription signals in BCSCs. These results show that the GR/YAP signaling pathway regulates BCSC formation and that caudatin may be a potential chemopreventive agent that targets breast cancer cells and CSCs.

Cynandione A (CA), isolated from ethyl acetate extract of Cynanchum wilfordii (CW), is a bioactive phytochemical that has been found to be beneficial for the treatment of several diseases. Hepatic de novo lipogenesis is one of the main causes of non-alcoholic fatty liver disease (NAFLD), which is thought to be a hepatic manifestation of certain metabolic syndromes. However, it has not yet been reported if CA has any therapeutic value in these diseases. Here, we investigated whether CA can inhibit hepatic lipogenesis induced by liver X receptor α (LXRα) using an in vitro model. We found that the extract and ethyl acetated layer of CW decreased the mRNA levels of sterol regulatory element-binding protein-1c (SREBP-1c), which plays a crucial role in hepatic lipogenesis. Additionally, we observed that CA could suppress the level of SREBP-1c, which was increased using two commercial LXRα agonists, GW3954 and T0901317. Moreover, the enzymes that act downstream of SREBP-1c were also inhibited by CA treatment. To understand the mechanism underlying this effect, the levels of phosphorylated AMP kinase (pAMPK) were measured after CA treatment. Therefore, CA might increase the pAMPK level by inducing phosphorylation of liver kinase B1 (LKB1), which can then convert AMPK to pAMPK. Taken together, we conclude that CA has an alleviative effect on hepatic lipogenesis through the stimulation of the LKB1/AMPK pathway.

This study aims to investigate alterations in volatile compounds and chemical components that may affect the sensory properties of Cynanchi wilfordii roasted at 120 °C and 180 °C for various durations of time (0 to 4 min). For volatile compound profiles, tridecane, tetradecane, and ethyl hexyl ether were detected as major compounds of the sample without roasting. As the roasting temperature increased, the types of volatile compounds increased, whereas the content decreased with an increase in heterocyclic compound generation. The sample roasted at 120 °C for 1 min showed a notably different profile pattern from the other roasted samples. For the sniffing test performed by trained panelists using gas chromatography olfactometry, naphthalene and nonanal, which induce spicy and raw C. wilfordii, respectively, were perceived most intensely throughout all the samples. In the electronic tongue analysis, the sample roasting at 180 °C showed an increased and decreased pattern in umami taste and saltiness, respectively, with increased roasting time. Electronic tongue analysis results showed that the sample roasted at 180 °C for 4 min would have the best sensory preference. Along with the volatile compound profiles, the sample roasted at 120 °C for 1 min had more sourness than the other samples, and would potentially have distinctive sensory characteristics. This study should provide baseline data on variations in the sensory properties and industrial applications of C. wilfordii upon roasting. PRACTICAL APPLICATION: Various applications of food ingredients through changes in sensory characteristics by roasting will provide consumers more options for food selections. Accordingly, this study will be utilized by (1) individuals and food companies who are interested in functional foods, (2) individuals and food industries that seek optimum roasting conditions for proper sensory characteristics of foods, and (3) farmers and crop producers who seek for utilization the applications of Cynanchi wilfordii as a food ingredient., (© 2019 Institute of Food Technologists®.)

Botanical-based natural products are an important resource for medicinal drug discovery and continue to provide diverse pharmacophores with therapeutic potential against cancer and other human diseases. A prototype Traditional Chinese Medicine (TCM) plant extract library has been established at the US National Cancer Institute, which contains both the organic and aqueous extracts of 132 authenticated medicinal plant species that collectively represent the potential therapeutic contents of most commonly used TCM herbal prescriptions. This library is publicly available in 96- and 384- well plates for high throughput screening across a broad array of biological targets, as well as in larger quantities for isolation of active chemical ingredients. Herein, we present the methodology used to generate the library and the preliminary assessment of the anti-proliferative activity of this crude extract library in NCI-60 human cancer cell lines screen. Particularly, we report the chemical profiling and metabolome comparison analysis of four commonly used TCM plants, namely Brucea javanica, Dioscorea nipponica, Cynanchum atratum, and Salvia miltiorrhiza. Bioassay-guided isolation resulted in the identification of the active compounds, and different extraction methods were compared for their abilities to extract cytotoxic compounds and to concentrate biologically active natural products., (Published by Elsevier B.V.)

Three new acetophenones, named cynwilforones A-C ( 1 - 3 ), together with cynandione A ( 4 ) were isolated from the root bark of Cynanchum wilfordii (Maxim.) Hemsl. Their structures were deduced based on spectroscopic analysis and chemical methods. Compounds 1 and 4 exhibited potential hypoglycemic effects through inhibition of hepatic gluconeogenesis by down-regulating the expression of phosphoenolpyruvate carboxykinase and glucose-6-phosphatase. This is the first report that acetophenones from the root bark of C. wilfordii possesses potential hypoglycemic activity in vitro .

A novel dimeric flavonol glycoside, Cynanflavoside A ( 1 ), together with six analogues, kaempferol-3- O - α -L-rhamnopyranosyl-(1→2)- β -D-glucopyranoside ( 2 ), quercetin-3- O - α -L-rhamnopyranosyl-(1→2)- β -D-glucopyranoside ( 3 ), kaempferol-3- O - α -L-rhamnopyranosyl-(1→2)- β -D-xylopyranoside ( 4 ), quercetin-3- O - α -L-rhamnopyranosyl-(1→2)- β -D-xylopyranoside (5) , kaempferol-3- O - β -D-glucopyranosyl-7- O - α -L-rhamnopyranoside ( 6 ), and quercetin-3- O -galactoside ( 7 ) were isolated from the n -butyl alcohol extract of Cynanchum acutum subsp. sibiricum . Their structures were determined spectroscopically and compared with previously reported spectral data. All compounds were evaluated for their anti-complementary activity in vitro , and only compound 5 exhibited anti-complement effects with CH 50 value of 0.33 mM.

The study aimed to investigate the mechanism of hepatoprotective effect of C-21 steroidal glucosides from Cynanchum auriculatum( Baishouwu) on oxidative stress in mice with liver injury. Mice were randomly divided into normal group,model group,positive control group,Baishouwu high group and Baishouwu low group. The liver injury model was induced by intraperitoneal injection of CCl4 peanut oil solution. All mice were sacrificed to collect blood and liver specimens. The activities of serum levels of ALT and AST were detected. The content of MDA and the activity of SOD in liver homogenate were examined by colorimetry method. Tissues were stained with hematoxylin-eosin for histological examination. The hepatic protein expressions of NF-κB p65,p-IκBα,i NOS and COX-2 were detected by Western blot. The mRNA expressions of TNF-α and IL-6 were determined by RT-PCR. It was found that treatment with C-21 steroidal glucosides from Baishouwu successfully attenuated liver injury induced by CCl4,as shown by decreased levels of serum biochemical indicators( AST,ALT)( P<0. 01). Administration of total C-21 steroidal glucosides enhanced the activity of SOD( P<0. 01) and decreased the content of MDA( P<0. 01) in liver homogenate. Microscopic features suggested that treatment with C-21 steroidal glucosides from Baishouwu was effective in inhibiting CCl4-induced hepatocyte edema and degeneration. Further studies showed that NF-κB p65 overexpression induced by CCl4 was decreased by C-21 steroidal glucosides,leading to the markedly down-regulated protein expression levels of p-IκBα,i NOS and COX-2,as well as the depression of TNF-α and IL-6 mRNA expressions. In conclusion,total C-21 steroidal glucosides from Baishouwu exhibited potent effect on oxidative stress pathway in mice with liver injury induced by CCl4,with enhanced activity of SOD,decreased content of MDA,and down-regulated levels of NF-κB p65,p-IκBα,i NOS and COX-2.

Seven undescribed C21 steroids, namely cynanchin A-G, together with thirteen known analogues, were isolated from the roots of cynanchum otophyllum. Their structures were elucidated by 1D, 2D NMR and MS spectra, as well as chemical methods. Meanwhile, all of isolates were tested for their anti-hepatic fibrosis activity. Among them, compounds 4-6, 10-12 and 14-17 showed moderate or significant inhibitory effects for the proliferation of hepatic stellate cells (HSCs) induced by transforming growth factor-β1 (TGF-β1) in vitro., (Copyright © 2019. Published by Elsevier B.V.)

Seven new polyhydroxypregnane glycosides, named cynotophyllosides P-V, together with three known analogs were isolated from the roots of Cynanchum otophyllum C.K.Schneid. Their structures were elucidated by a variety of spectroscopic techniques, as well as acid-catalyzed hydrolysis. All isolates were tested for their immunological activities in vitro against Con A- and LPS-induced proliferation of mice splenocytes. Immunoenhancing (for 1, 9) and immunosuppressive (for 2) activities were observed. Furthermore, cynotophylloside R (3) showed immunomodulatory as it enhanced the proliferation of splenocytes in low concentration and suppressed immune cells in concentration more than 1.0 μg/ml., (© 2019 Wiley-VHCA AG, Zurich, Switzerland.)

Oxidative stress plays an important role in the pathology of liver disorders. Total C‑21 steroidal glycosides (TCSGs), isolated from the root tuber of Cynanchum auriculatum Royle ex Wight, have been reported to exert numerous effects, including liver protective and antioxidant effects. In order to investigate the potential mechanisms underlying the protective effects of TCSGs on liver function, the present study used the human normal liver cell line, L02, to evaluate the effects of TCSGs on hydrogen peroxide (H2O2)‑induced oxidative injury and inflammatory responses. The L02 cells were pretreated with various concentrations of TCSGs, followed by exposure to 1.5 mM H2O2. Cell viability was determined by a 3‑(4,5‑dimethylthiazol‑2‑yl)‑2,5‑diphenyltetrazolium bromide (MTT) assay. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH) and nitric oxide (NO) were measured using colorimetric assays. The activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH‑Px) and the production of malondialdehyde (MDA) were also determined. Intracellular reactive oxygen species (ROS) levels were detected using a fluorescent probe. H2O2‑induced oxidative toxicity was attenuated following treatment with TCSGs, as indicated by the increase in cell viability, the decreased levels of ALT, AST, LDH, NO, MDA and ROS, and the increased activities of SOD, CAT and GSH‑Px. To further explore the possible mechanisms of action of TCSGs, the nuclear factor erythroid 2‑related factor 2 (Nrf2) and nuclear factor‑κB (NF)‑κB pathways were examined. The results revealed that treatment with TCSGs markedly induced Nrf2 nuclear translocation and upregulated the expression of heme oxygenase‑1 (HO‑1) in the L02 cells damaged by H2O2. In addition, pretreatment with TCSGs inhibited the NF‑κB signaling pathway by blocking the degradation of the inhibitor of nuclear factor κBα (IκBα), thereby reducing the expression and nuclear translocation of NF‑κB, as well as reducing the expression of tumor necrosis factor‑α (TNF‑α), interleukin-6 (IL‑6), inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX‑2). On the whole, the findings of this study demonstrate that TCSGs can protect L02 cells against H2O2‑induced oxidative toxicity and inflammatory injury by increasing the expression of Nrf2 and HO‑1, mediated by the NF‑κB signaling pathway.

In the present study, a novel polysaccharide fraction (CAP2-1) from Cynanchum auriculatum Royle ex Wight was obtained by hot water extraction, ethanol precipitation and sequential purification through anion-exchange and size-exclusion chromatography. CAP2-1 was a homogeneous heteropolysaccharide composed of mannose, rhamnose, glucuronic acid, galacturonic acid, galactose and arabinose, with an average molecular weight of 830.93 kDa. FT-IR and NMR spectra of CAP2-1 exhibited typical characteristic signals of polysaccharide. For antioxidant activity evaluation in vitro, CAP2-1 showed effective scavenging capacities against ABTS, DPPH and superoxide anion radical in a dose-dependent manner, with IC 50 values at 0.1232, 0.5543 and 0.5881 mg/mL, respectively. At cellular level, CAP2-1 provided a significant protective effect against hydrogen peroxide-induced oxidative stress in HepG2 cells by a compositive oxidation defense mechanism. CAP2-1 could reduce oxidative stress by significantly enhancing the contents of antioxidant enzyme SOD and non-enzymatic antioxidant GSH in oxidative damaged cells, in addition to scavenging ROS directly and improving cell viability and membrane integrity, consequently achieving the intracellular antioxidant activity. The results unveiled that CAP2-1 could be explored as a promising natural antioxidant for application in functional foods., (Copyright © 2018. Published by Elsevier B.V.)

As a part of our continuing research for bioactive constituents from Cynanchum limprichtii Schltr., two new C 21 steroidal glycosides limproside A (1) and limproside B (2) were isolated from the roots of Cynanchum limprichtii. Their structures were elucidated on the basis of 1D- and 2D-NMR spectroscopic data as well as HR-ESI-MS analysis. The cytotoxicity of two compounds against two selected human cancer cell lines was assayed.

Cynanchum wallichii Wight, is a traditional Chinese medicine herb, which is rich in saponins and has varieties of pharmacological activities. In this study, a standardized C. wallichii extract was established and the anti-tumor activity of the total saponins was evaluated by MTT assay. The extraction conditions of the standardized extract was optimized using response surface methodology. The experimental value was in good agreement (the yield 4.28%) with predicted values. The total saponins of the extract showed significant anti-tumor activity against three human tumor cell lines (A549, HepG2 and MCF-7), especially for MCF-7 (IC 50 . 67.63 μg/mL) cells in vitro.

Two new 13,14:14,15-disecopregnane-type C 21 steroidal glycosides, namely komarosides R (1) and S (2), along with four known compounds (3-6), were obtained from the 95% ethanol extract of the whole herbs of Cynanchum komarovii Al.Iljinski (Asclepiadaceae). The structures of new compounds were elucidated on the basis of 1D, 2D NMR spectroscopic data and acid hydrolysis. Compounds 1 and 2 showed potent inhibitory activities against human leukemia cell line (HL-60) with IC 50 values being 6.2 and 17.6 μM, respectively, compared to the positive control 5-fluorouracil (6.4 μM).

Introduction: Precise identification of botanical origin of plant species is crucial for the quality control of herbal medicine. In Korea, the root part of Cynanchum auriculatum has been misused for C. wilfordii in the herbal drug market due to their morphological similarities. Currently, DNA analysis using the polymerase chain reaction (PCR) method is employed to discriminate between these species., Objective: In order to develop a new analytical tool for the rapid discrimination of C. wilfordii and C. auriculatum, terahertz (THz) spectroscopy was employed., Methodology: Authentic samples of C. wilfordii and C. auriculatum were provided from the National Institute and standardized pellets for each species were prepared to get optimum results with terahertz time-domain spectroscopy (THz-TDS) in frequency range 0.2-1.20 THz., Results: The C. wilfordii pellet showed longer time delay compare to the sample of C. auriculatum and this was due to the difference in permittivity. The pellet samples of C. wilfordii and C. auriculatum showed a permittivity difference of about 0.08 at 0.2-1.20 THz., Conclusion: The experimental results indicated that THz-TDS analysis can be an effective and rapid method for the discrimination of C. wilfordii and C. auriculatum, and this application can be expanded for the discrimination of other similar herbal medicines., (Copyright © 2018 John Wiley & Sons, Ltd.)

The present study was designed to further investigate the C 21 steroidal glycosides in Cynanchum plants. Two new steroidal glycosides based on a 13, 14:14, 15-disecopregnane-type aglycone, komaroside P (1) and komaroside Q (2), together with three known compounds (3-5) were isolated from the whole herbs of Cynanchum komarovii. The aglycones of compounds 1 and 2 were two new disecopregnane. Their structures were elucidated on the basis of 1D, 2D NMR spectroscopic data and acid hydrolysis. All the compounds (1-5) showed potent inhibitory activities against human leukemia cell lines (HL-60) with IC 50 values ranging from 16.6 to 26.3 μmol·L -1 , compared to the positive control 5-fluorouracil (6.4 μmol·L -1 )., (Copyright © 2018 China Pharmaceutical University. Published by Elsevier B.V. All rights reserved.)

In this study, a novel method was developed to conjugate quercetin with Cynanchum auriculatum starch. Quercetin was first succinylized and then grafted onto C. auriculatum starch through a N,N'‑carbonyldiimidazole mediated reaction. The obtained water soluble quercetin grafted starch (quercetin-g-starch) was characterized by several instrumental methods. UV-vis spectrum of quercetin-g-starch aqueous solution exhibited two absorption bands at around 300 and 430nm. Fourier-transform infrared spectrum of quercetin-g-starch showed characteristic bands at 1731 and 1568cm -1 , assigning to CO stretching of esterified carboxyl group and CC stretching of aromatic ring, respectively. Proton nuclear magnetic resonance spectrum of quercetin-g-starch showed partial proton signals of starch (3.30-5.47ppm) and quercetin (6.19-7.68ppm). All above results suggested quercetin was successfully grafted onto C. auriculatum starch. Besides, quercetin-g-starch particles were irregular in shape and were in an amorphous state. Notably, the thermal stability, resistant starch content as well as antioxidant activity of C. auriculatum starch was greatly enhanced by grafting with quercetin. This indicated the potential of quercetin-g-starch in the development of a novel resistant starch with antioxidant activity., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Genus Cynanchum L. belongs to the family Asclepiadaceae, which comprise more than 200 species distributed worldwide. In Chinese medical practice, numerous drugs (such as tablets and powders) containing different parts of plants of this genus are used to treat snake bites, bruises, osteoblasts, rheumatoid arthritis and tumors. A search for original articles published on the cynanchum genus was performed by using several resources, including Flora of China Official Website and various scientific databases, such as PubMed, SciFinder, the Web of Science, Science Direct, and China Knowledge Resource Integrated (CNKI). Advances in the botanical, ethnomedicinal, phytochemical, and pharmacological studies of this genus are reviewed in this paper. Results showed that more than 440 compounds, including C21 steroids, steroidal saponins, alkaloids, flavonoids and terpene, have been isolated and identified from Cynanchum plants up to now. In vivo and in vitro studies have shown that plants possess an array of biological activities, including anti-tumor, neuroprotective and anti-fungal effects. Popular traditional prescription of Cynanchum sp. was also summed up in this paper. However, many Cynanchum species have received little or no attention. Moreover, few reports on the clinical use and toxic effects of Cynanchum sp. are available. Further attention should be focused on the study of these species to gather information on their respective toxicology data and relevant quality-control measures and clinical value of the crude extracts, active compounds, and bioactive metabolites from this genus. Further research on Cynanchum sp. should be conducted, and bioactivity-guided isolation strategies should be emphasized. In addition, systematic studies of the chemical composition of plants should be enhanced.

Eight new C21 steroidal glycosides, namely cynanotins A-H (1-8), together with fifteen known analogues, were isolated from the roots of Cynanchum otophyllum. Their structures were elucidated by spectroscopic analysis and chemical methods. In this study, all of isolates were tested for their vitro inhibitory activities against five human tumor cell lines (HL-60, SMMC-7721, A-549, MCF-7 and SW480). Compounds 3-15 showed moderate cytotoxic activities against HL-60 cell lines with IC 50 values ranging from 11.4 to 37.9 µM. Compounds 5, 9, and 10 showed marked or moderate cytotoxic activities against five human tumor cell lines with IC 50 values ranging from 11.4 to 36.7 µM. Compound 11 displayed moderate cytotoxic activities against HL-60, SMMC-7721, MCF-7 and SW480 cell lines with IC 50 values of 12.2-30.8 µM. Compared to the positive control (IC 50 : 35.0 µM), compounds 5, 9-11 exhibited more potential inhibitory activity against MCF-7 cells (IC 50 : 16.1-25.6 µM)., (Copyright © 2018. Published by Elsevier Ltd.)

Uncontrolled excessive activation of Hedgehog (Hh) signaling pathway is linked to a number of human malignant tumorigenesis. To obtain valuable Hh pathway inhibitors from natural product, in present study, a pair of novel epimers, Cynanbungeigenin C (CBC) and D (CBD) from the plant Cynanchum bungei Decne were chemically characterized by multiple spectroscopic data and chemical derivatization, and evaluated for their inhibition on Hh pathway. Mechanistically, CBC and CBD block Hh pathway signaling not through targeting Smo and Sufu, but at the level of Gli. In addition, both eipmers significantly suppress Hh pathway-dependent Ptch +/- ; p53 -/- medulloblastoma in vitro and in vivo. Furthermore, both CBC and CBD inhibited two Smo mutants induced Hh pathway activation, which suggested that they are potential compounds for the treatment of medulloblastoma with primary or acquired resistance to current Smo inhibitors. These results highlight the potential of CBC and CBD as effective lead compounds in the treatment of medulloblastoma and other Hh-dependent malignancy., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Bioassay-guided fractionation of the methanolic extract from the roots of Cynanchum atratum has resulted in the isolation of three new pregnane glycosides (1-3) along with four known compounds (4-7). Their structures were identified by analysis of the spectroscopic data including extensive 2D NMR. All of the isolates were evaluated for their potential to inhibit the melanin production in α-melanocyte stimulating hormone (α-MSH)-activated B16 melanoma cells. Of these, compounds 4-7 dose-dependently inhibited the melanin production with the IC 50 values ranging from 4 μM to 33 μM., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The herbaceous plant Cynanchum otophyllum Schneid. is widely used as a milk coagulant to make a Chinese traditional milk product, milk cake. However, the milk-clotting compounds and their mechanism remain unclear. In this study, crude proteases were extracted from the dried leaves of Cynanchum otophyllum Schneid. using citric acid-phosphate buffer and then partially purified by weak anion exchange chromatography. Two proteases, QA and QC, with molecular weights of 14 and 27 kDa, respectively, were shown to exhibit milk-clotting activity. A study of the effects of pH and temperature on the milk-clotting activity and proteolytic activity of the proteases showed that they exhibited good pH stability from pH 5.5 to 7.5 and good thermal stability at temperatures from 50 to 70°C. The QA and QC were the cysteine proteases, able to hydrolyze β-casein and κ-casein completely, and α-casein partially. The cleavage site on κ-casein determined by Orbitrap (Thermo Fisher Scientific, San Jose, CA) analysis showed that QA and QC could cleave κ-casein at Ser132-Thr133. Overall, the results suggest that the Cynanchum otophyllum Schneid. proteases are a promising milk-clotting enzyme that could be used for manufacturing milk cake and cheese., (Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)