Neriifolin

Yellow oleander (Thevetia peruviana) toxicosis in 4 goats.[Pubmed:37496389]

J Vet Diagn Invest. 2023 Sep;35(5):563-567.

Four alpine goats developed diarrhea soon after the owner placed plant clippings believed to be yellow oleander (Thevetia peruviana) into their pen on a suburban property near Palm Desert, CA, USA. A 1-y-old female goat died suddenly ~1 h after eating the plant clippings and was submitted to the San Bernardino Branch of the California Animal Health and Food Safety Laboratory System for postmortem examination. The main autopsy and histopathologic findings were myocardial hemorrhage and necrosis, consistent with cardiac glycoside intoxication. Rumen contents were analyzed by LC-MS/MS; peruvoside, a cardiac glycoside, was detected, but oleandrin, the cardiac glycoside of common oleander (Nerium oleander), was not. An LC-high-resolution MS (LC-HRMS) analysis revealed the presence of peruvoside and Neriifolin in the rumen contents and in a tested plant fragment, indicating that the plant was a member of the Thevetia genus. A clipping from the plant fed to the goats and submitted by the owner was identified as yellow oleander, Thevetia peruviana (also known as Cascabela thevetia).

Cardiac glycoside neriifolin exerts anti-cancer activity in prostate cancer cells by attenuating DNA damage repair through endoplasmic reticulum stress.[Pubmed:36792037]

Biochem Pharmacol. 2023 Mar;209:115453.

Prostate cancer (PCa) is one of the most common cancers in men. Patients with recurrent disease initially respond to androgen-deprivation therapy, but the tumor eventually progresses into castration-resistant PCa. Thus, new therapeutic approaches for PCa resistance to current treatments are urgently needed. Here, we report that cardiac glycoside Neriifolin suppresses the malignancy of cancer cells via increasing DNA damage and apoptosis through activation of endoplasmic reticulum stress (ERS) in prostate cancers. We found that cardiac glycoside Neriifolin markedly inhibited the cell growth and induced apoptosis in prostate cancer cells. Transcriptome sequence analysis revealed that Neriifolin significantly induced DNA damage and double strand breaks (DSBs), validated with attenuation expression of genes in DSBs repair and increasing phosphorylated histone H2AX (gamma-H2AX) foci formation, a quantitative marker of DSBs. Moreover, we found that Neriifolin also activated ERS, evidenced by upregulation and activation of ERS related proteins, including eukaryotic initiation factor 2alpha (eIF2alpha), protein kinase R (PKR)-like endoplasmic reticulum kinase (PERK) and C/EBP homologous protein (CHOP) as well as downregulation of CCAATenhancerbinding protein alpha (C/EBP-alpha), a transcriptional factor that forms heterodimers with CHOP. In addition, Neriifolin treatment dramatically inhibited the by tumor growth, which were reversed by CHOP loss or overexpression of C/EBP-alpha in nude mice. Mechanistically, Neriifolin suppressed the tumor growth by increasing DNA damage and apoptosis through CHOP-C/EBP-alpha signaling axis of ERS in prostate cancers. Taken together, these results suggest that cardiac glycoside Neriifolin may be a potential tumor-specific chemotherapeutic agent in prostate cancer treatment.

17beta-neriifolin from unripe fruits of Cerbera manghas suppressed cell proliferation via the inhibition of HOXA9-dependent transcription and the induction of apoptosis in the human AML cell line THP-1.[Pubmed:36266527]

J Nat Med. 2023 Jan;77(1):180-187.

Homeobox A9 (HOXA9) is a transcription factor that is overexpressed in acute myeloid leukemia (AML). It is associated with the pathogenesis and progression of AML, and is a factor responsible for a poor prognosis. Therefore, the development of HOXA9-targeting molecules may contribute to not only better understanding of the mechanism of HOXA9 regulation, but also the development of therapeutic applications. We constructed a reporter assay system using the promoter region of the KBTBD10 gene, to which HOXA9 directly binds and regulates transcription, in the human acute monocytic leukemia cell line THP-1. Using this luciferase gene assay, we screened 1120 plant extracts and a methanol extract of the unripe fruits of Cerbera manghas was found to suppress the reporter gene expression mediated by the KBTBD10 promoter. From the extract, five steroid-type compounds were identified as the active constituents: 7alpha-Neriifolin (1), 17beta-Neriifolin (2), 17alpha-digitoxigenin beta-D-glucosyl-(1 --> 4)-alpha-L-thevetoside (3), 17beta-digitoxigenin beta-D-glucosyl-(1 --> 4)-alpha-L-thevetoside (4), and acetylthevetin B (5). Among the five compounds, 17beta-Neriifolin most potently inhibited HOXA9-dependent gene expression without affecting the HOXA9 mRNA levels, and suppressed cell proliferation by inducing apoptosis. The findings on the structure-activity relationships of the compounds from C. manghas may contribute to the development of small molecule inhibitors of HOXA9.

Authentication of tejocote (Crataegus mexicana) dietary supplements based on DNA barcoding and chemical profiling.[Pubmed:34415825]

Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2021 Dec;38(12):1985-1994.

Tejocote (Crataegus mexicana, Mexican hawthorn), known as a weight-loss supplement, has been marketed online and is easily available for overseas direct purchase. Alipotec (brand name) is known as one of the most popular products containing tejocote in Mexico and other countries. However, adverse effects have been reported by users of these supplements. Therefore it is necessary to find the reason for the side effect. Dietary supplement samples labelled as containing tejocote were analysed using mass spectrometry and DNA barcoding analysis. Our results demonstrate that Alipotec samples contained ingredients from different species, yellow oleander instead of tejocote. The rpoB barcode region was able to differentiate between tejocote and yellow oleander species. Moreover, it was also observed that three compounds, including thevetin B, Neriifolin, and digitoxigenin, clearly distinguish between tejocote and yellow oleander samples. This is the first and preliminary investigation to use an integrated approach of both chemical and genomic profiling for the authentication of dietary supplement containing tejocote.

Investigation of cardiac glycosides from oleander in a human induced pluripotent stem cells derived cardiomyocyte model.[Pubmed:34371141]

Toxicol Lett. 2021 Oct 10;350:261-266.

The ingestion of Nerium oleander and Thevetia peruviana are common causes for poisoning in Southeast Asia. All parts of the oleander shrub contain cardiac glycosides of the cardenolide type. These glycosides act via inhibition of a Na(+)/K(+)-ATPase which might cause severe arrhythmia and subsequent death in oleander-poisoned patients. The current study uses human induced pluripotent stem cells derived cardiomyocytes (hiPSC-CM) in a microelectrode array (MEA) system to assess the cardiac effects of Neriifolin, oleandrin, digitoxigenin, peruvoside and thevetin A from the oleander plant. Digoxin was used as established reference compound. All tested compounds showed a corrected field potential duration (FPDc) shortening and was the lowest for 600 nM digitoxigenin with -36.9 +/- 1.2 %. Next to the dose-dependent pro-arrhythmic potential, a complete beat arrest of the spontaneously beating hiPSC-CM was observed at a concentration of 300 nM for Neriifolin, 600 nM for oleandrin and 1000 nM for digitoxigenin and peruvoside. Thevetin A did not cause arrhythmia up to a final concentration of 1000 nM. Thus, it was possible to establish a cardiac effect rank order of the tested substances: Neriifolin > oleandrin > digitoxigenin = peruvoside > digoxin > thevetin A.

The In Vitro Anti-Cancer Activities of 17betaH-Neriifolin Isolated from Cerbera odollam and its Binding Activity on Na+, K+-ATPase.[Pubmed:31530258]

Curr Pharm Biotechnol. 2020;21(1):37-44.

BACKGROUND: 17betaH-Neriifolin, a cardiac glycoside compound had been successfully isolated from Cerbera odollam leaves based on the bioassay guided-isolation procedure. The aim of these studies were to determine the in vitro anti-cancer and binding effects of 17betaH-Neriifolin on Na+, K+-ATPase. METHODS: The in vitro anti-cancer effects were evaluated using Sulphorhodamine B and Hoescht 33342 assays. The Na+, K+-ATPase assay was carried out using Malachite Green assay. In silico molecular docking studies and in vitro malachite green assay were used to predict the binding activities of 17betaH-Neriifolin on Na+, K+-ATPase and ouabain was also included as for comparison studies. RESULTS: The compound was tested against breast (MCF-7, T47D), colorectal (HT-29), ovarian (A2780, SKOV-3) and skin (A375) cancer cell lines that gave IC50 values ranged from 0.022 +/- 0.0015 to 0.030 +/- 0.0018 muM. The mechanism of cell death of 17betaH-Neriifolin was further evaluated using Hoescht 33342 assay and it was found that the compound killed the cancer cells via apoptosis. 17betaHNeriifolin and ouabain both bound at alpha-subunit in Na+, K+-ATPase and their binding energy were - 8.16 +/- 0.74 kcal/mol and -8.18 +/- 0.48 kcal/mol respectively. CONCLUSION: The results had confirmed the anti-proliferative effects exerted by 17betaH-Neriifolin in the breast, colorectal, ovarian and skin cancer cell lines. 17betaH-Neriifolin had shown to cause apoptotic cell death in the respective cancer cell lines.17betaH-Neriifolin and ouabain both bound at alpha-subunit in Na+, K+-ATPase and their binding energy were -8.16 +/- 0.74 kcal/mol and -8.18 +/- 0.48 kcal/mol respectively. This is the first report to reveal that 17betaH-Neriifolin managed to bind to the pocket of alpha-subunit of Na+.K+-ATPase.

Molluscicidal activity of cardiac glycosides isolated from Adenium obesum.[Pubmed:30838743]

Pest Manag Sci. 2019 Oct;75(10):2770-2775.

BACKGROUND: Terrestrial mollusks are one of most important agricultural pests worldwide. Natural phytochemicals have an extended history as a source of pesticides. This study was planned to isolate molluscicidal active compounds from the stems of Adenium obesum. RESULTS: The benzene-soluble fraction of the hydroethanolic extract displayed the most potent molluscicidal activity against Monacha obstructa among different solvent fractions with a median lethal dose (LD(50) ) of 4.91 microg g(-1) body weight (bw). The bioactivity-guided chemical exploration of the benzene-soluble fraction led to the isolation of two known cardiac glycosides, cerberin and Neriifolin which showed significant molluscicidal activity with LD(50) values of 5.39 and 4.3 microg g(-1) bw, respectively. CONCLUSION: Isolation of the cardiac glycoside Neriifolin from A. obesum and the molluscicidal activity of cerberin and Neriifolin against terrestrial snails are reported for the first time. (c) 2019 Society of Chemical Industry.

ATG-dependent phagocytosis in dendritic cells drives myelin-specific CD4(+) T cell pathogenicity during CNS inflammation.[Pubmed:29233943]

Proc Natl Acad Sci U S A. 2017 Dec 26;114(52):E11228-E11237.

Although reactivation and accumulation of autoreactive CD4(+) T cells within the CNS are considered to play a key role in the pathogenesis of multiple sclerosis (MS) and its animal model, experimental autoimmune encephalomyelitis (EAE), the mechanisms of how these cells recognize their target organ and induce sustained inflammation are incompletely understood. Here, we report that mice with conditional deletion of the essential autophagy protein ATG5 in classical dendritic cells (DCs), which are present at low frequencies in the nondiseased CNS, are completely resistant to EAE development following adoptive transfer of myelin-specific T cells and show substantially reduced in situ CD4(+) T cell accumulation during the effector phase of the disease. Endogenous myelin peptide presentation to CD4(+) T cells following phagocytosis of injured, phosphatidylserine-exposing oligodendroglial cells is abrogated in the absence of ATG5. Pharmacological inhibition of ATG-dependent phagocytosis by the cardiac glycoside Neriifolin, an inhibitor of the Na(+), K(+)-ATPase, delays the onset and reduces the clinical severity of EAE induced by myelin-specific CD4(+) T cells. These findings link phagocytosis of injured oligodendrocytes, a pathological hallmark of MS lesions and during EAE, with myelin antigen processing and T cell pathogenicity, and identify ATG-dependent phagocytosis in DCs as a key regulator in driving autoimmune CD4(+) T cell-mediated CNS damage.

Euphorbia neriifolia L.: Review on botany, ethnomedicinal uses, phytochemistry and biological activities.[Pubmed:28647179]

Asian Pac J Trop Med. 2017 May;10(5):430-438.

The present review is intended to provide information on botany, ethnomedicinal uses, phytochemistry and biological activities of various parts of Euphorbia neriifolia (E. neriifolia). E. neriifolia has several ethnomedicinal uses. The latex of E. neriifolia is used as laxative, purgative, rubefacient, carminative and expectorant as well as in treatment of whooping cough, gonorrhoea, leprosy, asthma, dyspepsia, jaundice, enlargement of the spleen, tumours, stone in the bladder, abdominal troubles and leucoderma. Leaves are brittle, heating, carminative, and good for improving the appetite and treatment of tumours, pains, inflammations, abdominal swellings and bronchial infections. Roots are used as symptomatic treatment of snake bite, scorpion sting and antispasmodic. Various plant parts or whole E. neriifolia extract and its isolates have been reported scientifically using various in-vivo and in-vitro experimental methods for anaesthetic, analgesic, anti-anxiety, anti-convulsant, anti-psychotic, anti-arthritis, anti-carcinogenic, antidiabetic, anti-diarrhoeal, anti-inflammatory, anti-thrombotic, antimicrobial, antioxidant, antiulcer, cytotoxic, death-receptor expression enhancing, dermal irritation, diuretic, haemolytic, immunomodulatory, radioprotective, scorpion venom and wound healing properties. It is reported to have chemical constituents like, Neriifolin-S, Neriifolin, neriifoliene, euphol, neriifolione, cycloartenol, nerifoliol, lectin, euphonerins A-G, 3-O-acetyl-8-O-tigloylingol, taraxerol, antiquorin, etc. Identified chemical constituents are still required to be explored for their advanced isolation techniques and biological activities.

The cardiac glycoside oleandrin induces apoptosis in human colon cancer cells via the mitochondrial pathway.[Pubmed:28597038]

Cancer Chemother Pharmacol. 2017 Jul;80(1):91-100.

PURPOSE: Evidence indicates that the cardiac glycoside oleandrin exhibits cytotoxic activity against several different types of cancer. However, the specific mechanisms underlying oleandrin-induced anti-tumor effects remain largely unknown. The present study examined the anti-cancer effect and underlying mechanism of oleandrin on human colon cancer cells. METHODS: The cytotoxicity and IC50 of five small molecule compounds (oleandrin, Neriifolin, strophanthidin, gitoxigenin, and convallatoxin) in human colon cancer cell line SW480 cells and normal human colon cell line NCM460 cells were determined by cell counting and MTT assays, respectively. Apoptosis was determined by staining cells with annexin V-FITC and propidium iodide, followed by flow cytometry. Intracellular Ca(2+) was determined using Fluo-3 AM,glutathione (GSH) levels were measured using a GSH detection kit,and the activity of caspase-3, -9 was measured using a peptide substrate. BAX, pro-caspase-3, -9, cytochrome C and BCL-2 expression were determined by Western blotting. RESULTS: Oleandrin significantly decreased cell viabilities in SW480, HCT116 and RKO cells. The IC50 for SW480 cells was 0.02 microM, whereas for NCM460 cells 0.56 microM. More interestingly, the results of flow cytometry showed that oleandrin potently induced apoptosis in SW480 and RKO cells. Oleandrin downregulated protein expression of pro-caspase-3, -9, but enhanced caspase-3, -9 activities. These effects were accompanied by upregulation of protein expression of cytochrome C and BAX, and downregulation of BCL-2 protein expression in a concentration-dependent manner. Furthermore, oleandrin increased intracellular Ca(2+) concentration, but decreased GSH concentration in the cells. CONCLUSIONS: The present results suggest that oleandrin induces apoptosis in human colorectal cancer cells via the mitochondrial pathway. Our findings provide new insight into the mechanism of anti-cancer property of oleandrin.

Antifertility activity of Thevetia peruviana (Pers.) K. Schum leaf in female Sprague-Dawley rat.[Pubmed:28066105]

Indian J Pharmacol. 2016 Nov-Dec;48(6):669-674.

OBJECTIVES: Thevetia peruviana (Pers.) K. Schum. (Apocynaceae) is known to possess cardioactive glycoside such as thevetin A, thevetin B, Neriifolin, peruvoside, thevetoxin, and ruvoside. Traditionally, T. peruviana leaves are used as abortifacient. The aim of the present study is to evaluate antifertility potential of T. peruviana leaves. SUBJECTS AND METHODS: Cardiac glycoside freed leaves of T. peruviana were extracted with methanol using maceration method. The dried cardiac glycoside-free methanolic extract of T. peruviana leaves (TPL-Me-G) was screened for phytoconstituents and evaluated for its effect on estrogen-primed female Sprague-Dawley rat uterus model. It was further studied for effects on the estrous cycle, implantation, and effect on estrogen and progesterone. STATISTICAL ANALYSIS USED: Statistical analysis was done by ANOVA followed by Dunnett's t-test. RESULTS: Alkaloids, flavonoids, essential oils, carbohydrates, and amino acids were found to be present in the glycoside-free extract. Thin-layer chromatography (TLC) in n-butanol: acetone: water (4:1:5) revealed the presence of quercetin and kaempferol. The presence of flavonoids (quercetin 0.0326% and kaempferol 0.138% on dry weight basis) was reconfirmed by high-performance TLC analysis. The extract was able to induce uterine contractions (EC(50), 0.170 mg/ml) in a dose-dependent manner. Further investigation showed significant (P < 0.001) extension of estrous cycle and anti-implantation activity of the extract by reduction of the progesterone level. CONCLUSIONS: Methanolic extract of T. peruviana leaves (TPL-Me-G) containing quercetin 0.0326% and kaempferol 0.138% possesses a significant (P < 0.001) antifertility potential by virtue of decreasing the progesterone level.

Acaricidal activity against Panonychus citri and active ingredient of the mangrove plant Cerbera manghas.[Pubmed:25918788]

Nat Prod Commun. 2014 Sep;9(9):1265-8.

Cerbera manghas is a mangrove plant which possesses comprehensive biological activities. A great deal of research has been undertaken on the chemical constituents and medical functions of C. manghas; insecticidal and antifungal activities have also been reported, but the acaricidal activity has not been studied. In our study, the acaricidal activity and active substances of C. manghas were investigated using a spray method, which showed that the methanol extracts of the fruit, twigs and leaves exhibited contact activity against female adults of Panonychus citri, with LC50 values at 24 h of 3.39 g L(-1), 4.09 g L(-1) and 4.11 g L(-1), respectively. An acaricidal compound was isolated from C. manghas by an activity-guided isolation method, and identified as (-)-17beta-Neriifolin, which is a cardiac glycoside. (-)-17beta-Neriifolin revealed high contact activity against female adults, nymphae, larvae and eggs of P. citri, with LC50 values at 24 h of 0.28 g L(-1), 0.29 g L(-1), 0.28 g L(-1) and 1.45 g L(-1), respectively.

Screening approach by ultra-high performance liquid chromatography-tandem mass spectrometry for the blood quantification of thirty-four toxic principles of plant origin. Application to forensic toxicology.[Pubmed:25438245]

J Chromatogr B Analyt Technol Biomed Life Sci. 2015 Jan 15;975:65-76.

Plant poisonings have left their mark on history and still cause many deaths, whether intentional or accidental. The means to show toxicological evidence of such poisonings should be implemented with great care. This article presents a technique for measuring thirty-nine toxic principles of plant origin in the blood, covering a large amount of toxins from local or exotic plants: alpha-lobeline, alpha-solanine, aconitine, ajmaline, atropine, brucine, cephalomannine, colchicine, convallatoxin, cymarine, cytisine, digitoxin, digoxin, emetine, gelsemine, ibogaine, jervine, kavain, lanatoside C, lupanine, mitragynine, Neriifolin, oleandrin, ouabain, paclitaxel, physostigmine, pilocarpine, podophyllotoxin, proscillaridin A, reserpine, retrorsine, ricinine, scopolamine, senecionine, sparteine, strophanthidin, strychnine, veratridine and yohimbine. Analysis was carried out using an original ultra-high performance liquid chromatography separation coupled with tandem mass spectrometry detection. Extraction was a standard solid phase extraction performed on Oasis((R)) HLB cartridge. Thirty-four of the thirty-nine compounds were put through a validation procedure. The assay was linear in the calibration curve range from 0.5 or 5 mug/L to 1000 mug/L according to the compounds. The method is sensitive (LOD from 0.1 to 1.6 mug/L). The within-day precision of the assay was less than 22.5% at the LLOQ, and the between-day precision was less than 21.5% for 10 mug/L for all the compounds included. The assay accuracy was in the range of 87.4 to 119.8% for the LLOQ. The extraction recovery and matrix effect ranged from 30 to 106% and from -30 to 14%, respectively. It has proven useful and effective in several difficult forensic cases.

Mechanisms of action of 17betaH-neriifolin on its anticancer effect in SKOV-3 ovarian cancer cell line.[Pubmed:25075041]

Anticancer Res. 2014 Aug;34(8):4141-51.

AIM: Abnormalities in apoptotic signalling pathways often occur in cancer cells and limit the successful chemotherapy outcomes in cancers. Therefore, there is an urgent need to discover new anticancer agents with novel mechanisms of action to overcome the resistance effect in chemotherapy. MATERIALS AND METHODS: In the present study, the anticancer effects and the mechanisms of action of 17betaH-Neriifolin (cardiac glycoside) were evaluated by terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay and a proteomic approach in treated and non-treated SKOV-3 ovarian cancer cells. RESULTS: 17betaH-Neriifolin was found to be active with IC50 values of 0.01 +/- 0.001 in SKOV-3 ovarian cancer cell line, as evaluated by the sulforhodamine B (SRB) assay. RESULTS from TUNEL assay indicated that 17betaH-Neriifolin caused apoptosis in SKOV-3 cells in a dose-dependent manner. Based on differential analysis of treated and non-treated SKOV-3 two-dimensional electrophoresis (2-DE) profiles, four proteins, namely vimentin (VIM), pyruvate kinase, muscle (PKM), heterogeneous nuclear ribonucleoprotein A1 (HNRNPA1) and transgelin (TAGLN1) were identified to be involved in apoptosis. Other proteins including piggybac transposable element derived 5 (PGBD5), DENN/MADD domain containing 2D (DENND2D) and formin-like 1(FMNL) have also been identified to be associated in SKOV-3 cell death induced by 17betaH-Neriifolin. CONCLUSION: These findings may provide new insights on the potential of 17betaH-Neriifolin's mechanism of action in killing ovarian cancer cells.

The principal toxic glycosidic steroids in Cerbera manghas L. seeds: identification of cerberin, neriifolin, tanghinin and deacetyltanghinin by UHPLC-HRMS/MS, quantification by UHPLC-PDA-MS.[Pubmed:24878878]

J Chromatogr B Analyt Technol Biomed Life Sci. 2014 Jul 1;962:1-8.

The toxicity of the sea mango (Cerbera manghas L.) is well known. The plant is ranked as one of the deadliest of the southern Asian coastline. Cardenolidic heterosides are responsible for the cardiotoxicity of trees of the Cerbera genus. We have identified and determined the concentration of the principal glycosidic steroids present in the seeds of sea mangos (Thailand). Drug screening of an extract of the seeds was carried out using ultra-high performance liquid chromatography coupled to photodiode array detection and mass spectrometry (UHPLC-PDA-MS) with quantification at 219nm. Identification was confirmed by UHPLC-HRMS. Deacetyltanghinin (m/z 549.3055+/-2ppm), Neriifolin (m/z 535.3259+/-2ppm), tanghinin (m/z 591.3169+/-2ppm) and cerberin (577.3375+/-2ppm) were the most abundant glycosidic steroids present in the sea mango seeds. A seed of the dried ripe fruit had concentrations of 1209.1, 804.2, 621.4 and 285.9mug/g, respectively. A seed of the fresh unripe fruit had concentrations of 49.4, 47.0, 3.5 and 2.3mug/g.