RESUMEN
Two series of fluoro substituted-anilino derivatives of naturally occurring hydroxybenzoquinone and hydroxynaphthoquinone were synthesized using TFA as catalyst to improve the product yield. Recently, fluorine containing compounds are being used as anticancer drugs. The aim of this study is to find compounds that are active against melanoma cells. This six new fluoro substituted quinone compounds were synthesized and characterized. All of these compounds were then subjected to molecular docking studies against B-raf protein using Discovery Studio 4.0 and the binding affinities were calculated. The energy scores of in silico analysis revealed that all the compounds exhibited better binding affinity towards B-raf protein. Moreover, all the derivatives and the parent compounds, embelin and plumbagin along with standard drug, PLX4032 were investigated for its in vitro cytotoxicity in A375 cell lines (Melanoma) and in vitro ELISA assay in B-raf isolated from melanoma cells. Among them, 5-(3-chloro-4-trifluoromethoxy-phenylamino)-2-hydroxy-3-undecyl-[1,4]benzoquinone exhibited lower cell viability with lowest LC50 of 12.25 µg/mL and thus poses suitability to be a lead molecule for further drug discovery.Communicated by Ramaswamy H. Sarma.
Asunto(s)
Antineoplásicos , Melanoma , Antineoplásicos/química , Línea Celular Tumoral , Proliferación Celular , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Simulación del Acoplamiento Molecular , Estructura Molecular , Relación Estructura-ActividadRESUMEN
BACKGROUND: Melanoma is one of the most common forms of skin cancer, and B-RAF is a mutated protein found in most melanomas. The important function of B-RAF is normal cell growth and survival. Most of the known B-RAF mutations are V600E mutations. Vemurafenib is the fluorine-based drug currently used for V600E mutations. However, this drug has side effects, therefore, more potent drugs with fewer side effects are required. OBJECTIVE: This study aims to develop a more effective lead compound as a B-RAF inhibitor from hydroxyquinone by structural modification of embelin, a naturally occurring hydroxybenzoquinone. It has the potency of detoxifying blood and is hence useful in a wide range of skin diseases. Thus, a fluorine substituted semisynthetic derivative of embelin, 5-(3-chloro-4-trifluoromethoxy phenyl amino)-2-hydroxy-3-undecyl- [1, 4] benzoquinone to fight against skin cancer was prepared. METHODS: Fluoro derivative of embelin was synthesized by the direct condensation of embelin with 3-chloro-4- trifluoromethoxy aniline. The structure of the product was characterized using various spectral data obtained from IR, 1H NMR, 19F NMR, 12C NMR, and mass spectrum. Various in vitro studies like antiproliferative study in A375 Cell Lines (B-RAF Elisa), western blotting analysis, gene expression study by reverse transcriptase PCR, caspase assay, flow cytometry analysis, clonogenic assay, and transwell migration assay were carried out to find its biological activity. RESULTS: A semisynthetic derivative of Embelin 5-(3-Chloro-4-trifluoromethoxy phenyl amino)-2-hydroxy-3- undecyl- [1, 4] benzoquinone (EOCF) was prepared, and the structure of the derivative was confirmed by spectral analysis. The MTT assay proves that the fluoro derivative of embelin exhibited better anti-cancer activity in melanoma cell lines than the parent compound, embelin. Western blot analysis showed that B-RAF expression level was reduced by the addition of derivative than the parent compound embelin. The Caspase ELISA analysis indicated that the derivative was found to be a good apoptotic marker. From the flow cytometry analysis, it was observed that cell arrest occurs at the G0/G1 phase. Its antimetastatic activity was determined using clonogenic assay. It indicated that the derivative EOCF inhibits the metastatic effects in melanoma cell lines. The migratory potential of melanoma cells was significantly reduced in the presence of EOCF when the transwell migration assay was conducted. CONCLUSION: This work established that the potency of the synthesized compound was more than the parent compound, embelin, when it was structurally modified with 3-chloro-4-trifluoromethoxy aniline. The derivative can be used as a lead molecule for further drug discovery.
Asunto(s)
Antineoplásicos/farmacología , Benzoquinonas/farmacología , Melanoma/tratamiento farmacológico , Inhibidores de Proteínas Quinasas/farmacología , Proteínas Proto-Oncogénicas B-raf/antagonistas & inhibidores , Antineoplásicos/síntesis química , Antineoplásicos/química , Benzoquinonas/síntesis química , Benzoquinonas/química , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Melanoma/metabolismo , Melanoma/patología , Estructura Molecular , Inhibidores de Proteínas Quinasas/síntesis química , Inhibidores de Proteínas Quinasas/química , Proteínas Proto-Oncogénicas B-raf/metabolismoRESUMEN
A phenaxazone compound [5H-Benzo[a]phenoxazin-5-one (BP)] along with an aminoquinone[2-[(o-hydroxyphenyl)amino]-1,4-naphthaquinone (HAN)] derivatives were synthesized from lawsone using ultrasound irradiation technique. The structure of the compounds were characterized by elemental analysis and various spectral studies. Optoelectronic properties were studied using Schrodinger material science suit (2015). The compounds exhibit fluorescence emission in longer wave length it may find applications in photodynamic therapy. Single crystal X-ray diffraction studies reveals that the compound BP crystallizes in monoclinic space group. The antioxidant activity of HAN and BP were determined using DPPH radical scavenging assay and the results indicate that both the compounds have good antioxidant capacity, HAN having more scavenging activity than BP. Lead molecules were identified using in silico molecular docking studies as a green chemistry approach. iGEMDOCK, GOLD and Schrödinger softwares were used for these studies. The docking studies reveal that the structural modification of the parent compound gave more active compounds making them promising lead molecules. The lead molecules were subjected to in vitro studies. The cytotoxicity of BP and HAN was studied using human breast cancer (SKBR3) cell lines. The IC50 value of HAN was found to be 19.8µM while BP was found to have cell viability, less than 10% even at 25µM concentration. The chemotherapeutic agents kill the cancer cells mainly through apoptosis. HAN and BP were subjected to apoptosis studies. BP was found to more active than HAN. Thus it can be suggested that the mechanism of cell death may be through apoptosis.
Asunto(s)
Apoptosis/efectos de los fármacos , Benzoxazinas/química , Benzoxazinas/farmacología , Muerte Celular/efectos de los fármacos , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Cristalografía por Rayos X , Humanos , Simulación del Acoplamiento Molecular , Naftoquinonas/química , Naftoquinonas/farmacología , Relación Estructura-ActividadRESUMEN
A simple and efficient procedure was employed for the synthesis of N'-(1,4-naphtho-quinone-2-yl) isonicotinohydrazide (NIH) by the reaction of 2-hydroxy-1,4-naphthaquinone (lawsone) and isonicotinoyl hydrazine in methanol using ultrasonic irradiation. Lawsone is the principal dye, isolated from the leaves of henna (Lawsonia inermis). Structural modification was done on the molecule aiming to get a more active derivative. The structure of the parent compound and the derivative was characterized by elemental analyses, infrared, electronic, (1)H, (13)C NMR and GC-MS spectra. The fluorescence spectral investigation of the compound was studied in DMSO and ethanol. Single crystal X-ray diffraction studies reveal that NIH crystallizes in monoclinic space group. The DNA cleavage study was monitored by gel electrophoresis method. The synthesized compound was found to have significant antioxidant activity against DPPH radical (IC50=58 µM). The in vitro cytotoxic studies of the derivative against two human cancer cell lines MCF-7 (human breast cancer) and HCT-15 (human colon carcinoma cells) using MTT assay revealed that the compound exhibited higher cytotoxic activity with a lower IC50 value indicating its efficiency in killing the cancer cells even at low concentrations. These results suggest that the structural modifications performed on lawsone could be considered a good strategy to obtain a more active drug.