Bufotenidine

Indolealkylamines in the venom of the scorpion Thorellius intrepidus.[Pubmed:37536653]

Toxicon. 2023 Sep;233:107232.

Scorpions are a group of arthropods that strike fear in many people due to their severe medical symptoms, even death, caused by their venomous stings. Even so, not all scorpion species contain harmful venoms against humans but still have valuable bioactive molecules, which could be used in developing new pharmaceutical leads for treating important diseases. This work conducted a comprehensive analysis of the venom from the scorpion Thorellius intrepidus. The venom of T. intrepidus was separated by size exclusion chromatography, and four main fractions were obtained. Fraction IV (FIV) contained small molecules representing over 90% of the total absorbance at 280 nm. Analysis of fraction FIV by RP-HPLC indicated the presence of three main molecules (FIV.1, FIV.2, and FIV.3) with similar UV absorbance spectra profiles. The molecular masses of FIV.1, FIV.2, and FIV.3 were determined, resulting in 175.99, 190.07, and 218.16 Da, respectively. Further confirmation through (1)H-NMR and (13)C-NMR analyses revealed that these molecules were serotonin, N-methylserotonin, and Bufotenidine. These intriguing compounds are speculated to play a pivotal role in self-defense and increasing venom toxicity and could also offer promising biotechnological applications as small bioactive molecules.

Two new polyamine alkaloids from the Bufo viridis toad venom.[Pubmed:35675547]

Nat Prod Res. 2023 Oct-Nov;37(20):3538-3542.

Two new polyamine alkaloids (bufonines A-B), together with four known alkaloids, Bufotenidine (3), bufotenine (4), 1-(beta-d-ribofuranosyl)-lH-1,2,4-triazone (5) and proline (6) were isolated from the Bufo viridis toad venom. Their structures were identified by UV, HR-ESI-MS, NMR spectral analyses, and comparison of theoretical and experimental ECD data. All compounds were tested in vitro cytotoxicity against three human cancer cell lines (HT-29, A549 and Hela). None of the compounds showed cytotoxicity towards all tested cell lines. To the best of our knowledge, this is the first report of alkaloid components from Bufo viridis toad venom.

[Study on anti-inflammatory activity and mechanism of indolealkylamines in toad skin on LPS-activated neutrophils].[Pubmed:34581088]

Zhongguo Zhong Yao Za Zhi. 2021 Sep;46(18):4774-4781.

Indolealkylamines(IAAs) are the main hydrophilic substances in toad skin, mainly including free N-methyl-5-hydroxytryptamine, bufotenine, Bufotenidine, dehydrobufotenine, and binding bufothionine. In this study, the LPS-activated neutrophils were used to investigate the structure-activity relationship and anti-inflammatory mechanism of the above-mentioned five monomers from the toad skin in vitro. The neutrophils were divided into the control group, model group(1 mug.mL~(-1) LPS), positive drug group(100 mug.mL~(-1) indometacin), as well as the low-(50 mug.mL~(-1)), medium-(100 mug.mL~(-1)) and high-dose(200 mug.mL~(-1)) free N-methyl-5-hydroxytryptamine, bufotenine, Bufotenidine, dehydrobufotenine, and binding bufothionine groups. The levels of IL-6, TNF-alpha and IL-1beta in the neutrophil supernatant of each group was measured by enzyme-linked immunosorbent assay(ELISA) after LPS stimulation, followed by the detection of apoptosis in each group after Annexin V/PI staining. The protein expression levels of caspase-3, Bax, Bcl-2, beclin1, LC3-I, and LC3-Ⅱ were assayed by Western blot. The results showed that IAAs reduced the excessive secretion of inflammatory cytokines caused by LPS compared with the model group. Besides, the activity of each free IAAs(N-methyl-5-hydroxytryptamine, bufotenine, Bufotenidine and dehydrobufotenine), especially bufotenine, was stronger than that of the binding bufothionine. As revealed by Annexin V/PI staining, LPS delayed the early apoptosis of neutrophils compared with the control group, while bufotenine promoted the apoptosis of neutrophils in a dose-dependent manner, which might be related to the elevated expression of apoptosis-related protein Bax/Bcl-2. In addition, LPS activated the autophagy pathways in neutrophils. This study confirmed the efficacy of IAAs in reducing the secretion of inflammatory cytokines in neutrophils induced by LPS for the first time. For instance, bufotenine exerts the anti-inflammatory effect possibly by inducing the apoptosis of neutrophils.

2,5-Di-methyl-bufo-tenine and 2,5-di-methyl-bufo-teni-dine: novel derivatives of natural tryptamines found in Bufo alvarius toads.[Pubmed:33614152]

Acta Crystallogr E Crystallogr Commun. 2021 Jan 29;77(Pt 2):190-194.

The solid-state structure of the bufotenine derivative bis-(5-meth-oxy-2,N,N-tri-methyl-tryptammonium) (5-MeO-2-Me-DMT) fumarate (systematic name: bis-[2-(5-meth-oxy-2-methyl-1H-indol-3-yl)eth-yl]di-methyl-aza-nium (2E)-but-2-enedioate), 2C(14)H(21)N(2)O(+).C(4)H(2)O(4) (2-), the Bufotenidine derivative 5-meth-oxy-2,N,N,N-tetra-methyl-tryptammonium (5-MeO-2-Me-TMT) iodide systematic name: [2-(5-meth-oxy-2-methyl-1H-indol-3-yl)eth-yl]tri-methyl-aza-nium iodide, C(15)H(23)N(2)O(+).I(-), and the hydrate of the same systematic name: [2-(5-meth-oxy-2-methyl-1H-indol-3-yl)eth-yl]tri-methyl-aza-nium iodide monohydrate, C(15)H(23)N(2)O(+).I(-).H(2)O, are reported. The structure of 5-MeO-2-Me-DMT fumarate possesses one tryptammonium cation and a half of a fumarate dianion in the asymmetric unit, linked together by N-Hcdots, three dots, centeredO hydrogen bonds in infinite two-dimensional networks parallel to the (101) plane. The structure of 5-MeO-2-Me-TMT iodide possesses one tryptammonium cation and one iodide anion in the asymmetric unit. The ions are linked via N-Hcdots, three dots, centeredI hydrogen bonds, and indoles are coupled in dimers through pi-pi inter-actions. The hydrate of 5-MeO-2-Me-TMT iodide possesses one tryptammonium cation, one iodide anion and one water mol-ecule in the asymmetric unit. It shows N-Hcdots, three dots, centeredI and O-Hcdots, three dots, centeredI hydrogen bonds that couple the tryptammonium cations into dimers.

Evaluation of analgesic and anti-inflammatory actions of indolealkylamines from toad venom in mice using lipidomics and molecular docking.[Pubmed:33321188]

J Ethnopharmacol. 2021 Apr 6;269:113677.

ETHNOPHARMACOLOGICAL RELEVANCE: Toad venom is one of widely used traditional Chinese medicines due to its analgesic and anti-inflammatory activities. However, hydrophilic alkaloids from toad venom, which may have certain pharmacological activities, have not been systematic studied. AIM OF THE STUDY: The aim of the study was to identify the indolealkylamines (IAAs) from toad venom and investigate the analgesic and anti-inflammatory actions. MATERIALS AND METHODS: The alkaloids were extracted and identified by high-resolution mass spectrometry. The analgesic abilities were determined using hot-plate test, formalin test and von Frey test. High-sensitivity lipidomics was used to investigate the regulatory function of IAAs on inflammatory eicosanoids. Besides, network pharmacology and molecular docking were used to demonstrate the candidate targets of IAAs. RESULTS: 22 constituents have been characterized by high performance liquid chromatography (HPLC)-Triple TOF 5600, including six specific IAAs (serotonin, N-methyl serotonin, bufotenine, Bufotenidine, bufothionine and dehydrobufotenine). Pharmacological studies showed that the IAAs from toad venom exerted significant analgesic activities at doses of 5, 15 and 45 mg/kg in vivo. Moreover, lipids analysis revealed IAAs might down-regulate inflammatory mediators from COX, LOX, DHA and LA pathways in formalin models, thus showing anti-inflammatory effect. The potent pharmacological function might because of the binding of IAAs and protein targets, such as sigma-1 receptor. CONCLUSION: The studies provided a systemic evidence for the analgesic and anti-inflammatory activities of IAAs from toad venom. It suggested that IAAs might be a potential candidate to reduce inflammatory pain disorders.

[Quality evaluation of different Berberidis Cortex species based on ~1H-NMR metabolomics and anti-diabetic activity].[Pubmed:33164432]

Zhongguo Zhong Yao Za Zhi. 2020 Oct;45(19):4677-4685.

To evaluate the quality differences of four mainstream species of Berberidis Cortex,~1H-NMR metabolomics was applied to detect its primary and secondary metabolites, and the partial least squares discriminant analysis and analysis of variance were integrated to screen differential metabolites between species. Furthermore, diabetic rat model was established by high fat diet and streptozotocin to assess differences in antidiabetic activities among the four species. Sixteen compounds were simultaneously detected and identified, including alkaloids, organic acids, carbohydrates and amino acids. Interspecific difference markers were revealed as magnoflorine, jateorhi-zine, Bufotenidine and saccharose for the first time. Berberis vernae and B. kansuensis presented superior activities on reducing blood glucose level, improving insulin resistance, increasing insulin sensibility and anti-inflammation. B. dictyophylla showed moderate antidiabe-tic effect, while B. diaphana rendered inferior antidiabetic capacity. Based on the contents of four differential markers and the results of antidiabetic activity evaluation, the quality of four Berberidis Cortex species was ranked as B.vernae approximately B.kansuensis>B. dictyophylla>B. diaphana. These results provided references for species collation, quality standard establishment and exploitation of Berberidis Cortex. The antidiabetic activities of B. vernae and B. kansuensis as well as their mechanisms of action merit further study in the future.

Two New Indole Alkaloids from Toad Venom of Bufo bufo gargarizans.[Pubmed:33019706]

Molecules. 2020 Oct 1;25(19):4511.

Two new indole alkaloids, Bufotenidine B (2) and Bufocarboline A (6), along with seven known indole alkaloids (1, 3-5, and 7-9) and three organic acids (10-12), were isolated from the water extract of toad venom. The structures of the new alkaloids were elucidated by extensive spectroscopic methods. The absolute configurations of 4, 6, and 8 were determined for the first time by electronic circular dichroism (ECD) calculations. The cytotoxic activity of all compounds was tested against human malignant melanoma cells A375 by the MTT method, and no antitumor activity was observed.

(1)H NMR-Based Metabolomics Coupled With Molecular Docking Reveal the Anti-Diabetic Effects and Potential Active Components of Berberis vernae on Type 2 Diabetic Rats.[Pubmed:32636751]

Front Pharmacol. 2020 Jun 19;11:932.

The dried stem bark of Berberis vernae C.K.Schneid., known as "Xiao-bo-pi" in Chinese, is a representative anti-diabetic herb in traditional Tibetan medical system. However, its anti-diabetic mechanisms and active components remain unclear. In this study, (1)H NMR-based metabolomics, biochemistry assay, molecular docking, and network analysis were integrated to evaluate the anti-diabetic effects of B. vernae extract on type 2 diabetic rats, and to explore its active components and underlying mechanisms. Diabetes was induced by high-fat diet and streptozotocin. After 30 days of treatment, B. vernae extract significantly decreased the serum levels of fasting blood glucose, insulin, insulin resistance index, glycated serum protein, TNF-alpha, IL-1beta, and IL-6, whereas significantly increased the serum levels of insulin sensitivity index in type 2 diabetic rats. A total of 28 endogenous metabolites were identified by (1)H NMR-based metabolomics, of which 9 metabolites that were changed by diabetes were significantly reversed by B. vernae extract. The constructed compound-protein-metabolite-disease (CPMD) interaction network revealed the correlation between chemical constituents, target proteins, differential metabolites, and type 2 diabetes. Ferulic acid 4-O-beta-D-glucopyranoside, Bufotenidine, jatrorrhizine, and berberine showed good hit rates for both the 30 disease-related proteins and 14 differential metabolites-related proteins, indicating that these four compounds might be the active ingredients of B. vernae against type 2 diabetes. Moreover, pathway analysis revealed that the anti-diabetic mechanisms of B. vernae might be related to its regulation of several metabolic pathways (e.g., butanoate metabolism) and disease-related signal pathways (e.g., adipocytokine signaling pathway). In summary, B. vernae exerts a significant anti-diabetic effect and has potential as a drug candidate for the treatment of type 2 diabetes.

[Determination of six compounds in Berberidis Cortex and comparative study of its different species].[Pubmed:30989857]

Zhongguo Zhong Yao Za Zhi. 2019 Mar;44(5):968-974.

In order to clarify the characteristic components of Berberidis Cortex,the preparative liquid chromatography and spectral analysis methods were used to separate and identify the unknown components in the water extract of Berberidis Cortex. Two compounds were isolated and identified as Bufotenidine and ferulic acid 4-O-beta-D-glucopyranoside. They were both isolated for the first time from Berberidis Cortex and Berberis. In addition,an HPLC method was successfully established for simultaneously determination of six compounds in Berberidis Cortex,and chemometric methods were used to study the chemical differences among three main species of Berberidis Cortex. The results suggested that jatrorrhizine and Bufotenidine are the main difference compounds among the three species.Compared with B. kansuensis and B. diaphana,B. vernae contains significantly more jatrorrhizine(P<0. 01),and the content of Bufotenidine in B. vernae was significantly higher than that in B. kansuensis(P<0. 05). Considering these results,further research is necessary to reveal the pharmacological activities of Bufotenidine and the pharmacodynamic differences between the three species. The results could provide a reference for quality control,the basic research on effective substances,and development of Berberidis Cortex.

N-methyl serotonin analogues from the Bufo bufo toad venom interact efficiently with the alpha7 nicotinic acetylcholine receptors.[Pubmed:28421441]

Dokl Biochem Biophys. 2017 Jan;472(1):52-55.

Two low-molecular-weight compounds were isolated from the parotid gland secret of the toad Bufo bufo, which by absorption spectra and HPLC-MS/MS chromatography data correspond to di- and trimethyl derivatives of serotonin (5-hydorxytryptamine): bufotenine (confirmed by counter synthesis) and Bufotenidine (5-HTQ). In experiments on competitive radioligand binding, these compounds showed a higher affinity and selectivity for neuronal alpha7 nicotinic acetylcholine receptors compared with the muscular cholinergic receptors. The most efficient compound in terms of binding value was bufotenine, the efficiency of 5-HTQ was an order of magnitude lower, and the minimal activity was exhibited by serotonin.

A New Indole Alkaloid from the Toad Venom of Bufo bufo gargarizans.[Pubmed:26999086]

Molecules. 2016 Mar 16;21(3):349.

A new indole alkaloid named bufobutarginine (1), along with three known bufotenines, namely, serotonin (2), Bufotenidine (3), and bufotenine (4), were isolated from the water extract of toad venom. Their structures were elucidated by spectral methods. This is the first time that arginine has been found to be involved in the biosynthesis of bufotenines in parotid of toad. The cytotoxic activities of these compounds have been assayed against A375 and A549 cell lines by the MTT method; however, they showed no cytotoxic activities.

Serotonin 5-O-beta-Glucoside and Its N-Methylated Forms in Citrus Genus Plants.[Pubmed:25893818]

J Agric Food Chem. 2015 Apr 29;63(16):4220-7.

Citrus genus is characterized by a specific presence of indole metabolites deriving from the N-methylation of tryptamine and its hydroxylated form, 5-hydroxytryptamine (serotonin), which are likely involved in plant defense mechanisms. In this study, we identified for the first time the occurrence in Citrus plants of serotonin 5-O-beta-glucoside and all its N-methylated derivatives, that is, N-methylserotonin 5-O-beta-glucoside, N,N-dimethylserotonin (bufotenine) 5-O-beta-glucoside, and N,N,N-trimethylserotonin (Bufotenidine) 5-O-beta-glucoside. The identification of the glucosylated compounds was based on mass spectrometric studies, hydrolysis by glucosidase, and in some cases, comparison to authentic compounds. Beside leaves, the distribution of the glucosylated forms and their aglycones in some Citrus species was evaluated in flavedo, albedo, juice, and seeds. The simultaneous presence of serotonin and its N-methylated derivatives, together with the corresponding glucosylated forms, is consistent with the occurrence of a metabolic pathway, specific for Citrus, aimed at potentiating the defensive response to biotic stress through the optimization of the production and use of the most toxic of such metabolites.

Citrus genus plants contain N-methylated tryptamine derivatives and their 5-hydroxylated forms.[Pubmed:23682903]

J Agric Food Chem. 2013 May 29;61(21):5156-62.

The occurrence and distribution in Citrus genus plants of N-methylated derivatives of tryptamine and their 5-hydroxylated forms are reported. Tryptamine, N-methyltryptamine, N,N-dimethyltryptamine, N,N,N-trimethyltryptamine, 5-hydroxytryptamine (serotonin), 5-hydroxy-N-methyltryptamine, 5-hydroxy-N,N-dimethyltryptamine (bufotenine), and 5-hydroxy-N,N,N-trimethyltryptamine (Bufotenidine) were quantitated by LC-ESI-MS/MS. Leaves of all citrus plants examined contained N,N,N-trimethyltryptamine, a compound that we first discovered in the bergamot plant. Interestingly, we also found out that all plants examined contained 5-hydroxy-N,N-dimethyltryptamine and 5-hydroxy-N,N,N-trimethyltryptamine, compounds never described so far in the Citrus genus. As N,N,N-trimethyltryptamine and 5-hydroxy-N,N,N-trimethyltryptamine possess nicotine-like activity by exerting their action on acetylcholine receptors, it is conceivable that both represent the arrival point of a biosynthetic pathway aimed to provide Citrus plants with chemical defense against aggressors. This hypothesis is supported by our finding that leaves and seeds, which are more frequently attacked by biotic agents, are the parts of the plant where the highest levels of those compounds were found.

Aeruginascin, a trimethylammonium analogue of psilocybin from the hallucinogenic mushroom Inocybe aeruginascens.[Pubmed:16673333]

Planta Med. 2006 Jun;72(7):665-6.

The hallucinogenic mushroom Inocybe aeruginascens contains several typical Psilocybe alkaloids including psilocybin. We have now elucidated the structure of a further indole derivative named aeruginascin as the quaternary ammonium compound N, N, N-trimethyl-4-phosphoryloxytryptamine. Aeruginascin is closely related to the frog skin toxin Bufotenidine (5-HTQ), a potent 5-HT3 receptor agonist, and has been found exclusively in Inocybe aeruginascens so far.

Utilization of O-phthalaldehyde-sulphuric acid as a spray reagent in thin-layer chromatographic detection of some indolealkylamines and application to cutaneous secretion extracts of toad species.[Pubmed:18965302]

Talanta. 1991 Nov;38(11):1303-7.

An expansion of the utilisation of o-phthalaldehyde in sulphuric acid medium as spray reagent was carried out when tryptophan and some tryptophan-derived indole alkylamines such as tryptamine, serotonin, bufotenine, dehydrobufotenine and Bufotenidine were examined by thin-layer chromatography. Rf-values and limits of detection ranging from 20 (serotonin) to 100 (dehydrobufotenine) ng per spot were found. Application of this reagent for the detection of some of these compounds was carried out, using either methanolic extracts or column chromatographic fractions of the skin secretion of the toads Bufo ictericus and Odontophrynus cultripes.