RESUMO
Cannabidiolic-acid (CBDA) synthase is the enzyme that catalyzes oxidative cyclization of cannabigerolic-acid into CBDA, the dominant cannabinoid constituent of the fiber-type Cannabis sativa. We cloned a novel cDNA encoding CBDA synthase by reverse transcription and polymerase chain reactions with degenerate and gene-specific primers. Biochemical characterization of the recombinant enzyme demonstrated that CBDA synthase is a covalently flavinylated oxidase. The structural and functional properties of CBDA synthase are quite similar to those of tetrahydrocannabinolic-acid (THCA) synthase, which is responsible for the biosynthesis of THCA, the major cannabinoid in drug-type Cannabis plants.
Assuntos
Cannabis/química , Cannabis/enzimologia , Oxirredutases Intramoleculares/genética , Oxirredutases Intramoleculares/fisiologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Cannabis/classificação , Cannabis/genética , Células Cultivadas , Clonagem Molecular , Coenzimas/metabolismo , Modelos Biológicos , Dados de Sequência Molecular , Estruturas Vegetais/química , Ligação Proteica , Spodoptera/genética , TransfecçãoRESUMO
To discover novel peroxisome proliferator-activated receptor gamma (PPARgamma) agonists that could be used as antidiabetic agents, we designed docosahexaenoic acid (DHA) derivatives (2 and 3), which have a hydrophilic substituent at the C4-position, based on the crystal structure of the ligand-binding pocket of PPARgamma. These compounds were synthesized via iodolactone as a key intermediate. We found that both DHA derivatives (2 and 3) showed PPARgamma transactivation higher than, or comparable to, that of pioglitazone, which is a TZD derivative used as an antidiabetic agent. DHA derivatives related to these potent compounds 2 and 3 were also synthesized to study structure-activity relationships. Furthermore, 4-OH DHA 2, which shows strong PPARgamma transcriptional activity, was separated as an optically pure form.
Assuntos
Ácidos Docosa-Hexaenoicos/síntese química , Ácidos Docosa-Hexaenoicos/farmacologia , Hipoglicemiantes/síntese química , Hipoglicemiantes/farmacologia , PPAR gama/agonistas , Animais , Células COS , Chlorocebus aethiops , Cristalografia por Raios X , Ácidos Docosa-Hexaenoicos/química , Desenho de Fármacos , Hipoglicemiantes/química , Espectroscopia de Ressonância Magnética , Espectrometria de Massas , Espectrofotometria UltravioletaRESUMO
Sphingosine-1-phosphate (S-1P) derivatives such as threo-(2S,3S)-analogues, which are C-3 stereoisomers of natural erythro-(2S,3R)-S-1P, have been synthesized starting from l-serine or (1S,2S)-2-amino-1-aryl-1,3-propanediols (6). threo-(1S,2R)-2-Amino-1-aryl-3-bromopropanols (HBr salt) have also been prepared from 6. The threo-S-1Ps and the threo-amino-bromide derivatives have shown potent inhibitory activity against Ca(2+) ion mobilization in HL60 cells induced by erythro-S-1P, suggesting that these compounds would compete with cell surface EDG/S1P receptors.