BFD-22 a new potential inhibitor of BRAF inhibits the metastasis of B16F10 melanoma cells and simultaneously increased the tumor immunogenicity.

Benzofuroxan is an interesting ring system, which has shown a wide spectrum of biological responses against tumor cell lines. We investigated, herein, the antitumor effects of benzofuroxan derivatives (BFDs) in vitro and in a melanoma mouse model. Cytotoxic effects of twenty-two BFDs were determined by MTT assay. Effects of BFD-22 in apoptosis and cell proliferation were evaluated using Annexin V-FITC/PI and CFSE staining. In addition, the effects in the cell cycle were assessed. Flow cytometry, western blot, and fluorescence microscopy analysis were employed to investigate the apoptosis-related proteins and the BRAF signaling. Cell motility was also exploited through cell invasion and migration assays. Molecular docking approach was performed in order to verify the BFD-22 binding mode into the ATP catalytic site of BRAF kinase. Moreover, the BFD-22 antitumor effects were evaluated in a melanoma murine model using B16F10. BFD-22 was identified as a potential hit against melanoma cells. BFD-22 induced apoptosis and inhibited cell proliferation of B16F10 cells. BFD-22 has suppressed, indeed, the migratory and invasive behavior of B16F10 cells. Cyclin D1 and CDK4 expression were reduced leading to cell cycle arrest at G0/G1 phase. Of note, phosphorylation of BRAF at Ser338 was strongly down-regulated by BFD-22 in B16F10 cells. The accommodation/orientation into the binding site of BRAF was similar of BAY43-9006 (co-crystallized inhibitor of BRAF, sorafenib). Importantly, BFD-22 presented in vivo antimetastatic effects and showed better therapeutic efficacy than sorafenib and taxol. BFD-22 can be considered as a new lead compound and, then, can be helpful for the designing of novel drug candidates to treat melanoma.

Planejamento, desenvolvimento e estudos de QSAR de derivados benzofuroxânicos com atividade frente Staphylococcus aureus e Trypanosoma cruzi / Design, development and QSAR studies of benzofuroxan derivatives with activity against Staphylococcus aureus and Trypanosoma cruzi

A modificação molecular de fármacos do arsenal terapêutico é estratégia promissora no planejamento e desenvolvimento de novas entidades químicas que possam apresentar características pertinentes deste fármaco, e suprimir suas características indesejáveis. Desta forma, na busca por novos compostos com atividade antimicrobiana, uma série de vinte [N'-(benzofuroxan-5-il)metileno]benzidrazidas substituídas, análogas funcionais da nifuroxazida (Passifuril®), foram sintetizadas e sua atividade biológica foi testada frente a cepas padrão e multirresistentes (MRSA e VISA) de Staphylococcus aureus, e frente a formas epimatigotas de Trypanosoma cruzi, agente causal da Doença de Chagas. A escolha dos grupos substituintes foi baseada em suas propriedades físico-químicas, tais como efeito eletrônico e hidrofobicidade, empregando o Diagrama de Craig. Os compostos foram obtidos por rota sintética descrita em literatura, assim como por rotas alternativas a fim de otimizar a metodologia tradicional e melhorar o rendimento dos produtos finais. Todos os compostos foram identificados e apresentam estrutura química inédita. A atividade dos vinte compostos frente S. aureus foi avaliada pelo método de determinação da concentração inibitória mínima (CIM); destes, dezesseis apresentaram os mesmos intervalos de CIM frente as cepas padrão e multirresistentes. O composto dissubstituído 3-CF3,4-NO2 (7t), apresentou a maior atividade com valores de CIM entre 12,7 - 11,4 µg/mL. A avaliação da atividade anti-T. cruzi também foi investigada, e na fase log de crescimento parasitário os compostos substituídos 4-H (7a), 4-CF3 (7n), 3,4-Cl2 (7s), 3-CF3,4-NO2 (7t) demonstraram os melhores resultados. O benznidazol, único fármaco utilizado no tratamento da Doença de Chagas, foi utilizado como referência nas mesmas concentrações. Os compostos que apresentaram melhores atividades nos ensaios realizados na fase estacionária de crescimento foram os compostos substituídos 4-I (7q) e 4-Br (7o) com valores de %IC50 de 6,11 µM e 7,38 µM, respectivamente. A influência das propriedades físico-químicas dos grupos substituintes em ambas as atividades foi observada e, a fim de avaliar quantitativamente suas contribuições para a bioatividade, estudos de QSAR-2D e QSAR-3D foram desenvolvidos, auxiliando assim na predição de novas estruturas com propriedades farmacológicas otimizadas, uma vez que os resultados obtidos indicam o forte potencial destes compostos na identificação de novos candidatos a fármaco antimicrobiano

Molecular modification of drugs from the therapeutic arsenal is a promising strategy for the design and development of new chemical entities that can demonstrate the relevant properties of this drug, and suppressing its undesirable properties. For the research of new leads with potential antimicrobial activity, a new series of twenty substituted [N´-(benzofuroxan-5-yl)methylene]benzohydrazides, nifuroxazide's (Passifuril®) functional analogs, was synthesized and tested against standard and multidrug-resistant Staphylococcus aureus (MRSA and VISA) strains and against epimastigote form of Trypanosoma cruzi, the etiological agent of Chagas' Disease. The selection of the substituent groups was based on their physicochemical properties, such as hydrophobicity and electronic effects, employing Craig's diagram. The designed compounds were obtained by synthetic route described in the literature, as well as by an alternative route, in order to optimize the traditional methodology and also to improve the final compounds yields. All compounds were identified as unpublished chemical structures. Bacterial activity of the twenty compounds against S. aureus was performed by minimal inhibitory concentration method (MIC), and sixteen of them exhibited similar bacteriostatic activity against standard and multidrug-resistant strains. The most active compound was the 3-CF3,4-NO2 disubstituted derivative (7t), which presented a MIC value from 12.7 to 11.4 µg/mL. Anti-T. cruzi activity was also investigated. The substituted compounds 4-H (7a), 4-CF3 (7n), 3,4-Cl2 (7s), 3-CF3,4-NO2 (7t) showed better results at logarithmic growth phase. Benznidazole, that is the only drug available to threat Chagas' disease, was used as a reference drug at the same concentrations of the compounds studied. The most effective substituded compounds were the 4-I (7q) and 4-Br (7o) substituted derivatives having %IC50 values of 6.11 µM and 7.38 µM, respectively, at stationary growth phase. The influence of the substituent's physicochemical properties on in vitro activities was observed, and, in order to establish quantitatively their contributions to bioactivity, 2D-QSAR and 3D-QSAR studies were developed, assisting in the prediction of new leads with improved pharmacological properties, since the results showed benzofuroxan derivatives as potential leads for identifying new drug candidates