RESUMO
Isofagomine (IFG) and its analogues possess promising glycosidase inhibitory activities. However, a flexible synthetic strategy toward both C5a-functionalized IFGs remains to be explored. Here we show a practical synthesis of C5a-S and R aminomethyl IFG-based derivatives via the diastereoselective addition of cyanide to cyclic nitrone 1. Nitrone 1 was conveniently prepared on a gram scale and in high yield from inexpensive (-)-diethyl D-tartrate via a straightforward method, with a stereoselective Michael addition of a nitroolefin and a Nef reaction as key steps. A 268-membered library (134 × 2) of the C5a-functionalized derivatives was submitted to enzyme- or cell-based bio-evaluations, which resulted in the identification of a promising ß-glucocerebrosidase (GCase) stabilizer demonstrating a 2.7-fold enhancement at 25 nM in p.Asn370Ser GCase activity and a 13-fold increase at 1 µM in recombinant human GCase activity in Gaucher cell lines.
RESUMO
The rapid discovery of a pharmacological chaperone toward human α-Gal A for the treatment of Fabry disease is described. Two polyhydroxylated pyrrolidines with the (3R,4S,5R) configuration pattern underwent rapid substituent diversity by conjugating the primary aminomethyl moiety of each with a variety of carboxylic acids to generate two libraries (2 × 60 members). Our bioevaluation results showed one member with the (2R,3R,4S,5R) configuration pattern and bearing a 5-cyclohexylpentanoyl group as a substituent moiety possessed sufficient chaperoning capability to rescue α-Gal A activity in the lymphocyte of the N215S Fabry patient-derived cell line and other α-Gal A mutants in COS7 cells.
Assuntos
Desenho de Fármacos , Doença de Fabry/tratamento farmacológico , Pirrolidinas/química , Pirrolidinas/farmacologia , Bibliotecas de Moléculas Pequenas/química , Bibliotecas de Moléculas Pequenas/farmacologia , Animais , Células COS , Chlorocebus aethiops , Estabilidade de Medicamentos , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , Inibidores Enzimáticos/uso terapêutico , Humanos , Hidroxilação , Cinética , Mutação , Pirrolidinas/síntese química , Pirrolidinas/uso terapêutico , Bibliotecas de Moléculas Pequenas/síntese química , Bibliotecas de Moléculas Pequenas/uso terapêutico , Estereoisomerismo , Temperatura , alfa-Galactosidase/antagonistas & inibidores , alfa-Galactosidase/genéticaRESUMO
A unique molecular library consisting of all sixteen synthetic ADMDP (1-aminodeoxy-DMDP) stereoisomers has been prepared and evaluated for inhibitory activity against α-Gal A, and ability to impart thermal stabilization of this enzyme. The results of this testing led us to develop a novel pharmacological chaperone for the treatment of Fabry disease. 3-Epimer ADMDP was found to be an effective pharmacological chaperone, able to rescue α-Gal A activity in the lymphoblast of the N215S Fabry patient-derived cell line, without impairment of cellular ß-galactosidase activity. When 3-epimer ADMDP was administered with rh-α-Gal A (enzyme replacement therapy) for the treatment of Fabry patient-derived cell lines, improvements in the efficacy of rh-α-Gal A was observed, which suggests this small molecule can also provide clinical benefit of enzyme replacement therapy in Fabry disease.
Assuntos
Terapia de Reposição de Enzimas/métodos , Doença de Fabry/tratamento farmacológico , Imino Açúcares/farmacologia , Pirrolidinas/farmacologia , alfa-Galactosidase/efeitos dos fármacos , Linhagem Celular Tumoral , Inibidores Enzimáticos/química , Estabilidade Enzimática/efeitos dos fármacos , Humanos , Imino Piranoses/química , Imino Piranoses/farmacologia , Imino Piranoses/uso terapêutico , Imino Açúcares/síntese química , Imino Açúcares/uso terapêutico , Manitol/análogos & derivados , Manitol/química , Manitol/farmacologia , Manitol/uso terapêutico , Pirrolidinas/síntese química , Pirrolidinas/uso terapêutico , Bibliotecas de Moléculas Pequenas/química , Bibliotecas de Moléculas Pequenas/farmacologia , EstereoisomerismoRESUMO
The asymmetric synthesis of the orthogonally protected N-mannosyl d-ß-hydroxyenduracididine (N-Man-d-ßhEnd) is described, starting from enantiopure silylated (S)-serinol. The key steps are: (i) glycosylamine formation between protected serinol and a benzylated d-mannose; (ii) guanidinylation; and (iii) cyclic guanidine formation. This synthesis constitutes a breakthrough in our studies towards a total synthesis of mannopeptimycin and should also allow for other studies in the field of mannopeptimycin research, including the synthesis of derivatives.
Assuntos
Produtos Biológicos/síntese química , Glicopeptídeos/síntese química , Manosídeos/síntese química , Pirrolidinas/síntese química , Produtos Biológicos/química , Glicopeptídeos/química , Manosídeos/química , Conformação Molecular , Pirrolidinas/química , EstereoisomerismoRESUMO
Preparation of Lipidâ II analogues containing an enzymatically uncleavable 1-C-glycoside linkage between the disaccharide moiety and the pyrophosphate- or pyrophosphonate-lipid moiety is described. The synthesis of a common 1-C-vinyl disaccharide intermediate has been developed that allows easy preparation of both an elongated sugar-phosphate bond and a sugar-phosphonate moiety, which are coupled with the polyprenyl phosphate to give the desired molecules. Inhibition studies show how a subtle structural modification results in dramatically different potency toward bacterial transglycosylase (TGase), and the results identify Lipidâ II-C-O-PP (IC50 =25 µM) as a potential TGase inhibitor.
Assuntos
Inibidores Enzimáticos/síntese química , Enzimas/química , Glicolipídeos/síntese química , Glicosiltransferases/química , Lipídeos/química , Monossacarídeos/química , Monossacarídeos/síntese química , Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , Glicolipídeos/química , Glicolipídeos/metabolismo , Glicosídeos , Glicosiltransferases/metabolismoRESUMO
A straightforward synthesis of novel, 2-heterocyclyl polyhydroxylated pyrrolidines is described. Stereocontrolled additions of nucleophiles to cyclic nitrones generated the corresponding 2,3-trans adducts, allowing the synthesis of the corresponding pyrrolidines via key intermediates bearing an alkyne and a nitrile oxide. Three hybrid systems, including a pyrrolidine with two isoxazoles and one triazole, are efficiently prepared via 1,3-dipolar cycloaddition. Biological testing of the product alkaloids showed that subtle structural variations have drastic effects on their inhibitory activities against glucosidases.
Assuntos
Produtos Biológicos/química , Inibidores Enzimáticos/química , Manosidases/antagonistas & inibidores , Pirrolidinas/química , Alcaloides/química , Alcaloides/metabolismo , Sítios de Ligação , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/metabolismo , Humanos , Cinética , Manosidases/metabolismo , Simulação de Acoplamento Molecular , Ligação Proteica , Estrutura Terciária de ProteínaRESUMO
The rapid discovery of ß-glucocerebrosidase (GCase) inhibitors and pharmacological chaperones for Gaucher disease is described. The N-aminobutyl DNJ-based iminosugar was synthesized and conjugating with a variety of carboxylic acids to generate a N-diversely substituted iminosugar-based library. Several members of this library were found to be nanomolar-range inhibitors of GCase; the inhibition constant Ki of the most potent was found to be 71nM. Although these new molecules showed reasonable chaperoning activity (1.5- to 1.9-fold) in the N370S fibroblast of Gaucher patient-derived cell line, this was accompanies by a concomitant decrease in the cellular α-glucosidase activity, which might limit their further therapeutic potential. Next, newly developed N-substituents were assembled with pyrrolidine-based scaffolds to generate new molecules for further evaluation. The new 2,5-dideoxy-2,5-imino-d-mannitol (DMDP)-based iminosugar 22 was found to exhibit a satisfactory chaperoning activity to enhance GCase activity by 2.2-fold in Gaucher N370S cell line, without impairment of cellular α-glucosidase activity.