digitoxigenin and periplogenin

A sensitive and specific ultra-performance liquid chromatographic-tandem mass spectrometric method was developed and validated to simultaneously determine periplocin, periplocymarin (PM), periplogenin (PG), periplocoside M (PSM) and periplocoside N (PSN) in rat plasma. Acetonitrile was employed to precipitate plasma with appropriate sensitivity and acceptable matrix effects. Chromatographic separation was performed using a Waters HSS T3 column with a gradient elution using water and acetonitrile both containing 0.1% formic acid and 0.1 mm ammonium formate within 8 min. Detection was performed in positive ionization mode using multiple reaction monitoring. The method was fully validated in terms of selectivity, linearity, accuracy, precision, recovery, matrix effects and stability. Using this method, the concentrations of periplocin, PM, PG, PSM and PSN were established after oral administration of Cortex Periplocae extract to rats, and the pharmacokinetic characteristics of periplocin, PM, PG, PSM and PSN were assessed. Generally, PM, PG, PSM and PSN were eliminated slowly and their half-lives were all >8 h. In addition, the systemic exposure of PSM showed significant differences between genders with more than 10 times higher area under the concentration-time curve in female rats than in male rats. The findings of this study provide useful information for further research on Cortex Periplocae.

Topics: Administration, Oral; Animals; Cardiac Glycosides; Chromatography, High Pressure Liquid; Digitoxigenin; Female; Male; Rats; Reproducibility of Results; Saponins; Tandem Mass Spectrometry

Periplogenin (PPG), a natural compound isolated from traditional Chinese herb Cortex Periplocae, has been reported to possess anti-inflammatory and anti-cancer properties.. The present study aims to investigate the antitumor effects of PPG and the underlying mechanism in human colorectal cancer cells.. The inhibition of cell growth in vitro was assessed by MTT assay. The induction of apoptosis and the ROS production induced by PPG was investigated by flow cytometry analysis. Western blotting was applied to measure the protein expression. Small interference RNA (siRNA) and a specific pharmacological inhibitor were used to knock down or inhibit the expression of related genes.. PPG was able to cause the production of ROS, inhibit the cancer cell growth and induce apoptosis. Nacetylcysteine was able to inhibit ROS production and apoptosis. PPG up-regulated the protein levels of BIP, peIF2α and CHOP as well as IRE1α and p-JNK, and down-regulated the protein level of p-ASK1, all of which were reversed by N-acetylcysteine. Importantly, knockdown of CHOP or JNK protein level attenuated the PPGelicited apoptosis.. PPG-induced apoptosis was regulated by ROS-mediated BIP/eIF2α/CHOP and BIP/ASK1/JNK signaling pathways in colon cancer cells, suggesting that PPG is a promising therapeutic agent for the treatment of human colon cancer.

Topics: Antineoplastic Agents; Apoptosis; Cell Line, Tumor; Cell Proliferation; Colonic Neoplasms; Digitoxigenin; Drug Discovery; Endoplasmic Reticulum; Endoribonucleases; Eukaryotic Initiation Factor-2; Gene Expression Regulation; Humans; MAP Kinase Kinase Kinase 5; MAP Kinase Signaling System; Periploca; Plant Extracts; Protein Serine-Threonine Kinases; Reactive Oxygen Species

The mortality rate of esophageal squamous cell carcinoma (ESCC) is higher than that of other cancers worldwide owing to a lack of therapeutic targets and related drugs. This study aimed to find new drugs by targeting an efficacious therapeutic target in ESCC patients. Signal transducer and activator of transcription 3 (STAT3) is hyperactive in ESCC. Herein, we identified a novel STAT3 inhibitor, periplogenin, which strongly inhibited phosphorylation of STAT3 at Tyr705. Docking models and pull-down assays revealed that periplogenin bound directly and specifically to STAT3, leading to significant suppression of subsequent dimerization, nuclear import, and transcription activities. In addition, STAT3 knockdown cell lines were insensitive to periplogenin, whereas in contrast, STAT3-overexpressing cells were more sensitive to periplogenin, indicating that STAT3 was a target of periplogenin. Intraperitoneally administered periplogenin exhibited efficacious therapeutic effects in ESCC patient-derived xenograft models and dramatically impaired the phosphorylation of STAT3 and expression levels of STAT3-mediated downstream genes. Thus, our study demonstrated that periplogenin acted as a new STAT3 inhibitor, suppressing the growth of ESCC in vitro and in vivo, providing a basis for its potential application in ESCC treatment and prevention.

Topics: Aged; Aged, 80 and over; Apoptosis; Cell Line, Tumor; Cell Proliferation; Cell Survival; Digitoxigenin; Esophageal Neoplasms; Esophageal Squamous Cell Carcinoma; Female; Humans; Male; Middle Aged; Phosphorylation; Signal Transduction; STAT3 Transcription Factor; Survival Rate; Xenograft Model Antitumor Assays

Periplogenin is a compound extracted from cortex periplocae. In the monomers' screening for inhibiting nasopharyngeal carcinoma, we found that periplogenin can inhibit nasopharyngeal carcinoma; however, its mechanism is still unclear. In this study, the chemical structure of periplogenin was uploaded to the PubChem database in order to obtain the target of periplogenin. The NPC's differential genes were obtained by analyzing the nasopharyngeal carcinoma data in the GEO database by R software. The common target of periplogenin and nasopharyngeal carcinoma was obtained through Cytoscape. Through R software analysis, ALB, epidermal growth factor receptor (EGFR), MAPK1, ESR1, MAPK8, SRC, CASP3, HSP90AA1, AR, MAPK14 may be the main targets of periplogenin in NPC. Through go enrichment analysis, it was found that periplogenin acted mainly on nasopharyngeal carcinoma through response to steroid metabolic process, cellular response to steroid hormone stimulus, hormone-mediated, and steroid hormone signaling pathway. After enrichment analysis on the Kyoto Encyclopedia of Genes and Genomes pathway, it was found that periplocan may inhibit NPC through the MAPK signaling pathway (the main signaling pathway), and the signaling pathway of proteoglycans in cancer, and the PI3K/AKT signaling pathway as well. In this study, we also carried out the experimental study of vitamin E together with periplogenin self-assembled nano-prodrugs in the treatment of NPC, and the results showed that tumor weight of PBS group was 0.531±0.039 g, while that of PPG group and MPSSV-NPs group was 0.258±0.059 g and 0.169±0.033 g, respectively, which was lower than PBS group. And the tumor inhibition rate of MPSSV-NPs was 69.41%, which was significantly higher than that of the PPG group (51.38%). This study demonstrated the mechanism of inhibition of nasopharyngeal carcinoma (NPC) by the monomer of periplogenin based on network pharmacology. We preliminarily confirmed that vitamin E coupled with a periplogenin self-assembled nano-prodrug has obvious effect in treating nasopharyngeal carcinoma.

Topics: Digitoxigenin; Humans; Nasopharyngeal Carcinoma; Nasopharyngeal Neoplasms; Phosphatidylinositol 3-Kinases; Prodrugs; Vitamin E

Periplocin is a cardiac glycoside and has been used widely in the clinic for its cardiotonic, anti-inflammatory and anti-tumor effects. Although it is taken frequently by oral administration in the clinic, there have been no reports demonstrating that periplocin could be detected in vivo after an oral administration, so there is an urgen need to determine the characteristics of periplocin in vivo after oral administration. In this study, a sensitive and reliable liquid chromatography-tandem mass spectrometry method was developed and validated to identify and quantify periplocin and its two metabolites in rat tissue after a single dosage of perplocin at 50 mg/kg. The results demonstrated that periplocin and its two metabolites were detected in all of the selected tissues; periplocin could reach peak concentration quickly after administration, while periplocymarin and periplogenin reached maximum concentration > 4.83 h after administration. The tissue distribution of analytes tended to be mostly in the liver, and higher analyte concentrations were found in the heart, liver, spleen, lung and kidney, but a small amount of chemical constituents was distributed into the brain. The consequences obtained using this method might provide a meaningful insight for clinical investigations and applications.

Topics: Animals; Cardiac Glycosides; Chromatography, Liquid; Digitoxigenin; Linear Models; Male; Rats; Rats, Sprague-Dawley; Reproducibility of Results; Saponins; Sensitivity and Specificity; Tandem Mass Spectrometry; Tissue Distribution

During a screening of Chinese plants traditionally used for the treatment of cancer and related diseases, extracts of the root bark of Periploca sepium Bunge showed strong cytotoxic activity.. Isolate and identify cytotoxic compounds from P. sepium and investigate the effects and mechanism of action on different cancer cell lines.. Extracts obtained with solvents of different polarities of the root bark of P. sepium were tested for their anti-proliferative effects. The most active extract was subjected to activity-guided fractionation using different chromatographic methods. The most active compound was further investigated on sarcoma cell lines regarding its effects concerning apoptosis, DNA damage and death receptor expression.. We isolated the cardiac glycosides periplocin, glucosyl divostroside, periplogenin, periplocymarin and periplocoside M with periplocin exhibiting the lowest IC. Periplocin displays a promising mechanism of action in sarcoma cells because altering the death receptor expression is an interesting target in sarcoma treatment especially to overcome TRAIL resistance.

Topics: Apoptosis; Cardiac Glycosides; Cell Line, Tumor; China; Digitoxigenin; Humans; Liposarcoma; Periploca; Plant Extracts; Plant Roots; Plants, Medicinal; Receptors, Death Domain; Saponins

Psoriasis is a multifactorial skin disease that inconveniences many patients. Considering the side effects and drug resistance of the current therapy, it is urgent to discover more effective and safer anti-psoriatic drugs. In the present study, we screened over 250 traditional Chinese medicine compounds for their ability to inhibit the cell viability of cultured human HaCaT keratinocytes, a psoriasis-relevant in vitro model, and found that periplogenin was highly effective. Mechanistic studies revealed that apoptosis and autophagy were not induced by periplogenin in HaCaT cells. However, periplogenin caused PI to permeate into cells, increased lactate LDH release and rapidly increased the number of necrotic cells. Additionally, the typical characteristics of necrosis were observed in the periplogenin-treated HaCaT cells. Notably, the necroptosis inhibitor Nec-1 and NSA were able to rescue the cells from necrotic cell death, supporting that necroptosis was involved in periplogenin-induced cell death. Furthermore, the ROS levels were elevated in the periplogenin-treated cells, NAC (an antioxidant) and Nec-1 could inhibit the ROS levels, and NAC could attenuate necroptotic cell death, indicating that the periplogenin-induced necroptotic cell death was mediated by oxidative stress. More importantly, in the murine models of TPA-induced epidermal hyperplasia and IMQ-induced skin inflammation, topical administration of periplogenin ameliorated skin lesions and inflammation. In sum, our results indicate, for the first time, that periplogenin is a naturally occurring compound with potent anti-psoriatic effects in vitro and in vivo, making it a promising candidate for future drug research.

Topics: Aminoquinolines; Animals; Cell Death; Cell Line; Cell Survival; Digitoxigenin; Disease Models, Animal; Female; Humans; Imiquimod; Mice; Mice, Inbred BALB C; Necrosis; Oxidative Stress; Psoriasis; Tetradecanoylphorbol Acetate

Periplogenin (PPG), a cardiac glycoside prepared from Cortex periplocae, with similar structure to bufalin, has been found to induce apoptosis in many tumor cells. However, lots of cardiac glycosides possessing strong antitumor activity in vitro have still not passed phase I clinical trials, mostly due to poor tumor selectivity and systemic toxicity. To overcome this drawback, we designed octreotide-periplogenin (OCT-PPG) conjugate by coupling PPG-succinate to the amino-terminal end of octreotide. In comparison with free PPG, the conjugate exhibited significantly stronger cytotoxicity on HepG2 cells (SSTRs overexpression) but much less toxicity in L-02 cells. After intravenous injection of OCT-PPG conjugate into H22 tumor-bearing mice, its total accumulation in tumor was 2.3 fold higher than that of free PPG, but was 0.71- and 0.84-fold lower in heart and liver, respectively, suggesting somatostatin-mediated target delivery of PPG into the tumor tissue and reduced distribution in heart and liver. In vivo studies using H22 tumor model in mice confirmed the remarkable therapeutic effect of this conjugate. These results suggested that OCT-PPG conjugate could provide a new approach for clinical application of cardiac glycosides and as a targeting agent for cancer therapy.

Topics: Animals; Antineoplastic Agents; Cell Line, Tumor; Cell Survival; Digitoxigenin; Drug Delivery Systems; Humans; Male; Mice; Neoplasms; Octreotide; Tissue Distribution; Tumor Burden

The seeds of Strophanthus kombé Oliv. are known to contain high levels of cardioactive compounds. However, the therapeutic use of Strophanthus in the treatment of cardiopathy requires more detailed knowledge of the compound profile to profit from the full potential of Strophanthus preparations. Therefore, the objective was to characterize the cardenolide profile and lipophilic constituents in S. kombé seeds using methods applicable in routine quality control. Freshly prepared S. kombé seed extracts were analyzed without previous sample clean-up using a novel HPLC-DAD-MSn method. In addition, seed oils were analyzed by GC-MS following derivatization of the lipids. More than 20 cardenolides were tentatively assigned in the seed extracts including strophanthidin, strophanthidol, periplogenin and strophanthidinic acid aglycones, carrying various saccharide moieties. The findings revealed the presence of eight novel cardenolides, which have not been described for S. kombé so far. The occurrence of strophanthidinic acid derivatives was verified by comparison with synthesized strophanthidinic acid-cymaropyranoside. GC-MS characterization of the oils mainly revealed the presence of fatty acids, especially oleic acid and linoleic acid, as well as phytosterols, the latter representing intermediates of cardenolide biosynthesis. In summary, these findings broaden our knowledge on the secondary metabolism of Strophanthus.

Topics: Cardiac Glycosides; Chromatography, High Pressure Liquid; Digitoxigenin; Fatty Acids; Gas Chromatography-Mass Spectrometry; Linoleic Acid; Lipids; Mass Spectrometry; Molecular Structure; Oleic Acid; Phytosterols; Plant Extracts; Seeds; Strophanthidin; Strophanthus

A method coupling high performance liquid chromatography with tandem mass spectrometry has been developed and validated for quantifying periplogenin in rat plasma using psoralen as an internal standard (IS). Plasma samples were pretreated using a simple liquid-liquid extraction with ethyl acetate and the chromatographic separation of periplogenin and psoralen was achieved on a Waters XBridge™ BEH C18 column with 0.1% formic acid and acetonitrile as mobile phase at a flow rate of 0.4mL/min. The detection was performed on a positive ion mode with electrospray ionization (ESI) source. The optimized ion transition pairs for quantitation were m/z 391.3→m/z 337.2 for periplogenin and m/z 187.0→m/z 131.0 for IS. The total run time was 9.0min. The calibration curve was linear over the range of 0.2-250ng/mL (r>0.99) with the lower limit of quantitation (LLOQ) at 0.2ng/mL. The intra- and inter-day precision were below 9.85% and the mean accuracy were from -10.03% to 10.26%. The average recoveries of periplogenin in plasma ranged from 85.1% to 95.6%. The proposed method was successfully applied in evaluating the pharmacokinetics of periplogenin after an oral dose of 30mg/kg Cortex Periplocae extract in rats.

Topics: Animals; Chromatography, Liquid; Digitoxigenin; Drug Stability; Linear Models; Male; Periploca; Plant Extracts; Rats; Rats, Sprague-Dawley; Reproducibility of Results; Sensitivity and Specificity; Tandem Mass Spectrometry

A sensitive and reliable LC-MS/MS method was developed and validated for the simultaneous determination of periplocin and its two metabolites (periplocymarin and periplogenin) in rat plasma using psoralen as the internal standard (IS). After liquid-liquid extraction with ethyl acetate, chromatographic separation was performed on a C18 column with a 13 min gradient elution using 0.1% formic acid and acetonitrile as mobile phase at a flow rate of 0.3 mL/min. The detection was accomplished on a tandem mass spectrometer via an electrospray ionization (ESI) source by multiple reaction monitoring (MRM) in the positive ionization mode. The lower limits of quantitation (LLOQs) for periplocin, periplocymarin and periplogenin were 0.5, 1 and 0.1 ng/mL, respectively. The mean recoveries of the analytes and IS were higher than 67.7%. The proposed method was successfully applied to evaluating the pharmacokinetic studies of periplocin and its metabolites (periplocymarin and periplogenin) in rats after a single oral administration of periplocin at 50 mg/kg.

Topics: Acetonitriles; Administration, Oral; Animals; Calibration; Cardiac Glycosides; Chromatography, Liquid; Digitoxigenin; Formates; Limit of Detection; Male; Plasma; Quality Control; Rats; Rats, Sprague-Dawley; Saponins; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry

The importance of glycoside in the regulation of thyroid dysfunction is not well understood. In the present investigation, effects of periplogenin-3- O-D-glucopyranosyl (1→6)(1→4)-D-cymaropyranoside, isolated from the vegetable, LAGENARIA SICERARIA, in L-thyroxine (L-T₄)-induced hyperthyroidism and in related cardiovascular abnormalities have been revealed in Wistar albino rats. L-T₄ (500 μg/kg, s. c./d) administration for 12 days significantly increased serum concentrations of thyroxine (T₄), triidothyronine (T₃), and hepatic 5'-deiodinase I (5'-DI) activity with a parallel increase in lipid peroxidation (LPO) in different organs such as heart, liver and kidney; serum glucose and insulin concentrations and a decrease in cardiac Na (+)-K (+)-ATPase activity as well as serum total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, very low-density lipoprotein cholesterol and triglycerides. Most of these adverse effects were reversed following the administration of isolated periplogenin. However, out of its 3 different concentrations (5.0, 10, and 25 mg/kg), 5 mg/kg appeared to be the most effective one as it could nearly normalize the level of T₃, glucose, insulin, Na (+)-K (+)-ATPase activity, tissue LPO and different serum lipids suggesting the protective role of periplogenin against thyrotoxicosis and associated cardiovascular problems. It appears that the periplogenin actions are mediated through its direct antithyroidal and/or LPO inhibiting properties.

Topics: Animals; Cardiovascular Diseases; Digitoxigenin; Humans; Hyperthyroidism; Lipid Peroxidation; Magnoliopsida; Plant Extracts; Random Allocation; Rats; Rats, Wistar; Thyroxine

In the present study, a 75% ethanol extract of Streptocaulon juventas (SJ), which had a strong inhibitory effect on the proliferation of human lung A549 adenocarcinoma cells, was subjected to bioassay-guided fractionation. The most active fraction (SJF) was obtained using a macroreticular resin column followed by a silica-gel column. Then its in vivo effect on lung cancer was investigated in athymic nude mice with A549 tumors while its effects on body weight, blood biochemical indicators, and organ indices were monitored. The results showed that SJF inhibited the tumor growth significantly at day 10 and day 15 during treatment without physical side effects. Following HPLC and NMR spectrometry, the main components of SJF were identified as digitoxigenin, periplogenin, and periplogenin glucoside.

Topics: Animals; Antineoplastic Agents, Phytogenic; Apocynaceae; Cell Line, Tumor; Digitoxigenin; Lung Neoplasms; Mice; Mice, Nude; Molecular Structure; Neoplasms, Experimental; Plant Extracts; Xenograft Model Antitumor Assays

Doxorubicin is a common chemotherapeutic anticancer drug. Its use is associated with adverse effects including cardiotoxicity. Several therapeutics interventions have been attempted to reduce the toxicity and to improve the efficacy of the drug. However, on phytochemicals very few investigations have been made. In the present study we have evaluated the potential of a cardenolide, periplogenin, isolated from the leaves of Aegle marmelos in protecting the doxorubicin induced cardiotoxicity and lipid peroxidation (LPO) in rats. Doxorubicin induced cardiac and hepatotoxicity were characterized by marked biochemical changes including an increase in serum creatine kinase-MB (CK-MB), glutamate-pyruvate transaminase (SGPT), and tissue LPO, with a decrease in superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH). It also increased the levels of different serum lipids, but decreased the amount of high-density lipoprotein (HDL). Cotherapy of the test cardenolide and doxorubicin for 4 weeks reversed all these adverse effects. However, out of three different concentrations (12.5, 25, and 50 mg/kg p.o.) of the test periplogenin, 25 mg/kg appeared to be most effective. When its efficacy was compared with that of vitamin E (alpha-tocopherol) the isolated compound exhibited a better therapeutic potential. The isolated periplogenin from the leaves of A. marmelos could potentially inhibit doxorubicin-induced cardiovascular problems in rats. However, its moderate dose was found to be most effective.

Topics: Aegle; Alanine Transaminase; alpha-Tocopherol; Animals; Antibiotics, Antineoplastic; Antioxidants; Body Weight; Cardiovascular Diseases; Catalase; Chemical and Drug Induced Liver Injury; Creatine Kinase, MB Form; Digitoxigenin; Disease Models, Animal; Dose-Response Relationship, Drug; Doxorubicin; Glutathione; Lipid Peroxidation; Lipids; Liver Diseases; Plant Leaves; Rats; Rats, Wistar; Superoxide Dismutase

To study the constituents of the stems of Periploca forrestii.. The compounds were separated and purified by silica gel column chromatography, recrystallization and high-performance liquid chromatography. The structures were identified by various spectroscopic methods.. Nine compounds were isolated and identified as 3-O-acetyloleanolic acid (1), 14-ursen-3-ol-1-one (2), taraxasterol (3), jacoumaric acid (4), periplogenin (5), 2alpha,3beta-dihydroxyursolic acid (6), E-p-hydroxy-cinnamic acid (7), caffeic acid (8), proanthocyanidin A2 (9).. All compounds except 6 were isolated from this plant for the first time, compound 4, 9 were obtained from the Periploca for the first time.

Topics: Caffeic Acids; Chromatography, High Pressure Liquid; Digitoxigenin; Magnetic Resonance Spectroscopy; Periploca; Proanthocyanidins; Sterols; Triterpenes

Endogenous Na(+)-pump specific inhibitors are present in the plasma, urine, and tissues of humans and animals. To date, the source of these inhibitors has not been rigorously defined. In the present study, large amounts of several Na(+)-pump specific inhibitors have been demonstrated to exist in the urine of rats raised on a regular chow diet and tap water. All of the inhibitor levels have been found to increase 1.5-8-fold by the surgical preparation of reduced renal mass (RRM) and one-kidney, one-clip (IK, IC) hypertension. These urinary inhibitors, however, except for the ouabain-like inhibitor which eluted from a high performance liquid chromatography C18 column at the same retention time as [3H]ouabain, disappeared within a week after switching the diet from regular diet (number 5001, PMI Feeds, Inc.) to pure synthetic diet (number 5755). The urinary level of the ouabain-like inhibitor decreased to only one-half of the level in the control rat raised on a regular diet. Two of these inhibitors were purified from both urine and diet by a combination of Amberlite XAD-2 adsorption chromatography, reverse phase low pressure liquid chromatography, and several high performance liquid chromatographies. Reverse phase high performance liquid chromatography, liquid secondary ion and gas-liquid mass spectrometries, and proton nuclear magnetic resonance spectroscopy identified these inhibitors as a stereoisomer of convalloside, probably neoconvalloside, and a mono-rhamnoside of periplogenin or its stereoisomer. These cardiac glycosides exhibited inhibitory potencies comparable to ouabain against ouabain-displacement from Na+,K(+)-ATPase and against 86Rb uptake into human erythrocytes, and they also exhibited cross-reactivity to anti-ouabain antibodies and anti-digoxin antibodies. These results clearly demonstrate that the principal source of most of the inhibitors in rat urine is the diet. The results suggest that the ouabain-like inhibitor may be derived from an endogenous origin.

Topics: Analysis of Variance; Animal Feed; Animals; Brain; Cardiac Glycosides; Cell Membrane; Diet; Digitoxigenin; Glycosides; Humans; Hypertension, Renovascular; Magnetic Resonance Spectroscopy; Male; Mass Spectrometry; Molecular Structure; Rats; Rats, Sprague-Dawley; Reference Values; Sodium-Potassium-Exchanging ATPase

Topics: Digitoxigenin

Topics: Digitoxigenin

Topics: Digitoxigenin; Digoxigenin; Glucosides

Topics: Digitoxigenin; Strophanthidin