Bufarenogin

Toxicon. 2016 Feb;110:27-34.

Bufadienolides from parotoid gland secretions of Cuban toad Peltophryne fustiger (Bufonidae): Inhibition of human kidney Na(+)/K(+)-ATPase activity.[Pubmed: 26615828 ]

Parotoid gland secretions of toad species are a vast reservoir of bioactive molecules with a wide range of biological properties.
METHODS AND RESULTS:
Herein, for the first time, it is described the isolation by preparative reversed-phase HPLC and the structure elucidation by NMR spectroscopy and/or mass spectrometry of nine major bufadienolides from parotoid gland secretions of the Cuban endemic toad Peltophryne fustiger: ψ-Bufarenogin, gamabufotalin, Bufarenogin, arenobufagin, 3-(N-suberoylargininyl) marinobufagin, bufotalinin, telocinobufagin, marinobufagin and bufalin. In addition, the secretion was analyzed by UPLC-MS/MS which also allowed the identification of azelayl arginine. The effect of arenobufagin, bufalin and ψ-Bufarenogin on Na(+)/K(+)-ATPase activity in a human kidney preparation was evaluated. These bufadienolides fully inhibited the Na(+)/K(+)-ATPase in a concentration-dependent manner, although arenobufagin (IC50 = 28.3 nM) and bufalin (IC50 = 28.7 nM) were 100 times more potent than ψ-Bufarenogin (IC50 = 3020 nM).
CONCLUSIONS:
These results provided evidence about the importance of the hydroxylation at position C-14 in the bufadienolide skeleton for the inhibitory activity on the Na(+)/K(+)-ATPase.

Biocatalysis, 2011, 29(2-3):96-101.

Biotransformation of arenobufagin and cinobufotalin by Alternaria alternata.[Reference: WebLink]

The biotransformation of arenobufagin (1) and cinobufotalin (2), isolated from the natural medicine Chan Su, by Alternaria alternata AS 3.4578 was carried out.
METHODS AND RESULTS:
Incubation of 1 and 2 afforded six metabolites: 3-oxo-arenobufagin (1a), ψ-Bufarenogin (1b), 3-oxo-ψ-Bufarenogin (1c), 3-oxo-Δ4-derivative of cinobufotalin (2a), 3-oxo-cinobufotalin (2b) and 12β-hydroxycinobufotalin (2c). Among them, metabolites 1a, 1c and 2c are new compounds and their structures were characterized on the basis of their spectroscopic data (NMR, MS and IR).
CONCLUSIONS:
Compounds 1, 2, 1b, 2a and 2b were evaluated for their cytotoxicity against HepG2 and MCF-7 human cancer cells, and all of them showed significant inhibitory activities.

Oncotarget, 2015, 6(13):11627-39.

ψ-Bufarenogin, a novel anti-tumor compound, suppresses liver cancer growth by inhibiting receptor tyrosine kinase-mediated signaling.[Pubmed: 25890498]

Resistance of hepatocellular carcinoma (HCC) to existing chemotherapeutic agents largely contributes to the poor prognosis of patients, and discovery of novel anti-HCC drug is in an urgent need.
METHODS AND RESULTS:
Herein we report ψ-Bufarenogin, a novel active compound that we isolated from the extract of toad skin, exhibited potent therapeutic effect in xenografted human hepatoma without notable side effects. In vitro, ψ-Bufarenogin suppressed HCC cells proliferation through impeding cell cycle progression, and it facilitated cell apoptosis by downregulating Mcl-1 expression. Moreover, ψ-Bufarenogin decreased the number of hepatoma stem cells through Sox2 depression and exhibited synergistic effect with conventional chemotherapeutics. Mechanistic study revealed that ψ-Bufarenogin impaired the activation of MEK/ERK pathway, which is essential in the proliferation of hepatoma cells. ψ-Bufarenogin notably suppressed PI3-K/Akt cascade, which was required in ψ-Bufarenogin-mediated reduction of Mcl-1 and Sox2. ψ-Bufarenogin inhibited the auto-phosphorylation and activation of epithelial growth factor receptor (EGFR) and hepatocyte growth factor receptor (c-Met), thereafter suppressed their primary downstream cascades Raf/MEK/ERK and PI3-K/Akt signaling.
CONCLUSIONS:
Taken together, ψ-Bufarenogin suppressed HCC growth via inhibiting, at least partially, receptor tyrosine kinases-regulated signaling, suggesting that ψ-Bufarenogin could be a novel lead compound for anti-HCC drug.

Yao Xue Xue Bao. 2014 Nov;49(11):1574-7.

[Chemical constituents of bufadienolides in cinobufacino injection].[Pubmed: 25757284]

Cinobufacino injection is purified from water extraction of the skin of Bufo bufo gargarizans, which has been widely used for various cancers in clinic with significant anti-tumor effects. Bufadienolides were regarded as the main active constituents of cinobufacino injection in previous reports.
METHODS AND RESULTS:
In present study, 6 bufadienolides were isolated and purified from Cinobufacino injection. Their structures were identified as 3-epi-ψ-Bufarenogin (1), ψ-Bufarenogin (2), 3-epi-arenobufagin (3), arenobufagin (4), 3-epi-gamabufotalin (5), and 3-oxo-arenobufagin (6), separately.
CONCLUSIONS:
Among them, 1 and 3 were new compounds, 5 and 6 were new natural products. Compounds 1, 2 and compounds 3, 4 were two pairs configuration isomers at C-3, separately.