title: Bufadienolides from parotoid gland secretions of Cuban toad Peltophryne fustiger (Bufonidae): Inhibition of human kidney Na(+)/K(+)-ATPase activity.
abstract: Parotoid gland secretions of toad species are a vast reservoir of bioactive molecules with a wide range of biological properties. 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). 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.
authors.affiliation: Faculdade de Farmácia, Universidade Federal do Rio de Janeiro, CCS, Bloco A,Ilha do Fundão, 21.941-590 Rio de Janeiro, Brazil. Electronic address: Wilmer.Perera@gmail.com.
authors.forename: Wilmer H
authors.initials: WH
authors.lastname: Perera Córdova
authors.affiliation: Faculdade de Farmácia, Universidade Federal do Rio de Janeiro, CCS, Bloco A,Ilha do Fundão, 21.941-590 Rio de Janeiro, Brazil.
authors.forename: Suzana Guimarães
authors.initials: SG
authors.lastname: Leitão
authors.affiliation: Laboratório de Farmacologia Bioquímica e Molecular, Instituto de Ciências Biomédicas, CCS Bloco J, Ilha do Fundão, 21941-902, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Geraldino
authors.initials: G
authors.lastname: Cunha-Filho
authors.affiliation: Facultad de Biología, Universidad de La Habana, Calle 25 No. 455, Vedado, Havana City, Cuba.
authors.forename: Roberto Alonso
authors.initials: RA
authors.lastname: Bosch
authors.affiliation: Facultad de Biología, Universidad de La Habana, Calle 25 No. 455, Vedado, Havana City, Cuba.
authors.forename: Isel Pascual
authors.initials: IP
authors.lastname: Alonso
authors.affiliation: Departamento de Farmacia, Facultad de Química, Universidad Nacional Autónoma de México, Ciudad Universitaria, Mexico City, 04510 DF, Mexico.
authors.forename: Rogelio
authors.initials: R
authors.lastname: Pereda-Miranda
authors.affiliation: Laboratório de Farmacologia Bioquímica e Molecular, Instituto de Ciências Biomédicas, CCS Bloco J, Ilha do Fundão, 21941-902, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Rodrigo
authors.initials: R
authors.lastname: Gervou
authors.affiliation: Laboratório de Farmacologia Bioquímica e Molecular, Instituto de Ciências Biomédicas, CCS Bloco J, Ilha do Fundão, 21941-902, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Natália Araújo
authors.initials: NA
authors.lastname: Touza
authors.affiliation: Laboratório de Farmacologia Bioquímica e Molecular, Instituto de Ciências Biomédicas, CCS Bloco J, Ilha do Fundão, 21941-902, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Luis Eduardo M
authors.initials: LE
authors.lastname: Quintas
authors.affiliation: Laboratório de Farmacologia Bioquímica e Molecular, Instituto de Ciências Biomédicas, CCS Bloco J, Ilha do Fundão, 21941-902, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: François
authors.initials: F
authors.lastname: Noël
mesh_terms: D000664:Amphibian Venoms / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism / Q000633:toxicity*; D000818:Animals; D002018:Bufanolides / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism / Q000633:toxicity*; D002025:Bufonidae / Q000254:growth & development / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D003462:Cuba; D006801:Humans; D006900:Hydroxylation; D007668:Kidney / Q000187:drug effects* / Q000201:enzymology; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D049990:Membrane Transport Modulators / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism / Q000633:toxicity*; D015394:Molecular Structure; D010306:Parotid Gland / Q000378:metabolism*; D045483:Rivers; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization; D018629:Spectrometry, Mass, Secondary Ion; D053719:Tandem Mass Spectrometry
chemical_list: D000664:Amphibian Venoms; D002018:Bufanolides; D049990:Membrane Transport Modulators; D000254:Sodium-Potassium-Exchanging ATPase
keywords: 1D and 2D NMR spectroscopy; Bufadienolides; HRESIMS; Na(+)/K(+) ATPase; Peltophryne fustiger; UPLC-MS/MS; Western Giant toad