Shahanavaj Khan

FGFR a promising druggable target in cancer: Molecular biology and new drugs

INTRODUCTION The Fibroblast Growth Factor Receptor (FGFR) family consists of Tyrosine Kinase Receptors (TKR) involved in several biological functions. Recently, alterations of FGFR have been reported to be important for progression and development of several cancers. In this setting, different studies are trying to evaluate the efficacy of different therapies targeting FGFR. AREAS COVERED This review summarizes the current status of treatments targeting FGFR, focusing on the trials that are evaluating the FGFR profile as inclusion criteria: Multi-Target, Pan-FGFR Inhibitors and anti-FGF (Fibroblast Growth Factor)/FGFR Monoclonal Antibodies. EXPERT OPINION Most of the TKR share intracellular signaling pathways; therefore, cancer cells tend to overcome the inhibition of one tyrosine kinase receptor by activating another. The future of TKI (Tyrosine Kinase Inhibitor) therapy will potentially come from multi-targeted TKIs that target different TKR simultaneously. It is crucial to understand the interaction of the FGF-FGFR axis with other known driver TKRs. Based on this, it is possible to develop therapeutic strategies targeting multiple connected TKRs at once. One correct step in this direction is the reassessment of multi target inhibitors considering the FGFR status of the tumor. Another opportunity arises from assessing the use of FGFR TKI on patients harboring FGFR alterations.

Optimizing indomethacin-loaded chitosan nanoparticle size, encapsulation, and release using Box–Behnken experimental design

Indomethacin chitosan nanoparticles (NPs) were developed by ionotropic gelation and optimized by concentrations of chitosan and tripolyphosphate (TPP) and stirring time by 3-factor 3-level Box-Behnken experimental design. Optimal concentration of chitosan (A) and TPP (B) were found 0.6mg/mL and 0.4mg/mL with 120min stirring time (C), with applied constraints of minimizing particle size (R1) and maximizing encapsulation efficiency (R2) and drug release (R3). Based on obtained 3D response surface plots, factors A, B and C were found to give synergistic effect on R1, while factor A has a negative impact on R2 and R3. Interaction of AB was negative on R1 and R2 but positive on R3. The factor AC was having synergistic effect on R1 and on R3, while the same combination had a negative effect on R2. The interaction BC was positive on the all responses. NPs were found in the size range of 321-675nm with zeta potentials (+25 to +32mV) after 6 months storage. Encapsulation, drug release, and content were in the range of 56-79%, 48-73% and 98-99%, respectively. In vitro drug release data were fitted in different kinetic models and pattern of drug release followed Higuchi-matrix type.

Synthesis and anti-Candidal activity of N-(4-aryl/cyclohexyl)-2-(pyridine-4-yl carbonyl) hydrazinecarbothioamide.

Eighteen N-(4-aryl/cyclohexyl)-2-(pyridine-4-yl carbonyl) hydrazinecarbothioamide derivatives were synthesized, evaluated against ten clinical isolates of Candida spp. and compared with itraconazole. Introduction of p-chloro (2c), p-iodo (2q), m-chloro (2l) and o-nitro (2r) substitution at phenyl ring of thiosemicarbazide enhanced the anti-Candida activity. Compound (2c) bearing p-cholorophenyl ring was found to be the most effective against Candida albicans ATCC 66027, Candida spp. 12810 (blood) and Candida spp. 178 (HVS) with MIC value of 0.09-0.78 μg/mL, whereas itraconazole exhibits the inhibitory activity with MIC value of 0.04-1.56 μg/mL against all tested strains. There is a correlation between anti-Candidal activity and p-chloro substitution at phenyl ring of thiosemicarbazide. All synthesized compounds were investigated for their potential cytotoxicity against non cancer cell line MCF-10A. The active compounds 2c, 2r and 2a were further investigated for their cytotoxic effects on three cancer cell lines; HT1080 (skin), HepG2 (liver) and A549 (lung). The active compounds showed minimal cytotoxic activity against non cancer cell line and all three cancer cell lines. Moreover, compound 2c displaying better activity against C. albicans ATCC66027 and Candida spp. [blood] compared to reference drug (itraconazole), represents a good lead for the development of newer, potent and broad spectrum anti-Candidal agents.

Potential role of Escherichia coli DNA mismatch repair proteins in colon cancer

The epithelium of gastrointestinal tract organizes many innate defense systems against microbial intruders such as integrity of epithelial, rapid eviction of infected cells, quick turnover of epithelial cell, intrinsic immune responses and autophagy. However, Enteropathogenic Escherichia coli (EPEC) are equipped with well developed infectious tricks that evade the host defense systems and utilize the gastrointestinal epithelium as a multiplicative foothold. During multiplication on and within the epithelium, EPEC secrete various toxins that can weaken, usurp, and use many host cellular systems. However, the possible mechanisms of pathogenesis are still poorly elusive. Recent study reveals the existence of EPEC in colorectal cancer patients and their potential role in depletion of DNA mismatch repair (MMR) proteins of host cell in colonic cell lines. The EPEC colonised intracellularly in colon mucosa of colorectal carcinoma whereas extracellular strain was detected in mucosa of normal colon cells. Interestingly, alteration in MutS, MutL complexes and MUTYH of mammalian cells may be involved in development of CRC. These data propose that MMR of E. coli may be potential therapeutic targets and early detection biomarkers for CRC. This article reviews the potential role of E. coli MutS, MutL and MutY protein in CRC aetiology.

Computational prediction of Mycoplasma hominis proteins targeting in nucleus of host cell and their implication in prostate cancer etiology.

Cancer has long been assumed to be a genetic disease. However, recent evidence supports the enigmatic connection of bacterial infection with the growth and development of various types of cancers. The cause and mechanism of the growth and development of prostate cancer due to Mycoplasma hominis remain unclear. Prostate cancer cells are infected and colonized by enteroinvasive M. hominis, which controls several factors that can affect prostate cancer growth in susceptible persons. We investigated M. hominis proteins targeting the nucleus of host cells and their implications in prostate cancer etiology. Many vital processes are controlled in the nucleus, where the proteins targeting M. hominis may have various potential implications. A total of 29/563 M. hominis proteins were predicted to target the nucleus of host cells. These include numerous proteins with the capability to alter normal growth activities. In conclusion, our results emphasize that various proteins of M. hominis targeted the nucleus of host cells and were involved in prostate cancer etiology through different mechanisms and strategies.

In vitro evaluation of anticancer and biological activities of synthesized manganese oxide nanoparticles

This paper presents the results from a systematic study into the characterization and anticancer and biological activity of synthesized super-paramagnetic manganese oxide nanoparticles (Mn3O4-NPs). The Mn3O4-NPs showed an IC50 value of 666.44 μg mL−1 on the HT29 human colorectal cell line. Furthermore, we investigated the molecular mechanism of Mn3O4-NPs for the inhibition of Bcl-2 and Bcl-xL through an in vitro study. Bcl-2 and Bcl-xL are key signaling regulators of the programmed cell death pathway and ensure proper apoptosis and have been proposed as a novel target for anticancer therapy. The down-regulation of Bcl-2 and Bcl-xL on the HT29 cancer cell line showed that the NPs under study could be useful in the treatment of cancer. The down-regulation of the anti-apoptotic regulators Bcl-2 and Bcl-xL enhances cytotoxicity in colon carcinoma cells connected with the induction of apoptosis. Moreover, the in vitro haemolysis study antibacterial tests showed that Mn3O4-NPs have no significant toxicity and antibacterial activity, thereby exhibiting their biocompatibility and reliability in biomedical sciences.

Gut Microbiota and Probiotics: Current Status and Their Role in Cancer Therapeutics

Preclinical Research

In vitro evaluation of cytotoxicity, possible alteration of apoptotic regulatory proteins, and antibacterial activity of synthesized copper oxide nanoparticles

Copper oxide nanoparticles (CuO-NPs) were synthesized using a urea-based thermal decomposition technique, and characterized using different techniques. X-ray diffraction (XRD) and Raman spectroscopy confirmed the phase purity and crystalline structure of CuO-NPs. The size of CuO-NPs was investigated using XRD and was confirmed via dynamic light scattering analysis. CuO-NPs showed an average diameter of ∼20nm. The possible cytotoxicity of CuO-NPs was evaluated in HT-29 and SW620 cancer cell lines. The median inhibitory concentration of CuO-NPs in HT-29 and SW-620 cells was 4.99 and 3.75μg/mL, respectively. The underlying mechanism responsible for apoptosis in colon cancer cells after CuO-NP exposure has not been well understood. In this study, we investigated the possible mechanisms of induction of apoptosis via analysis of the expression of Bcl-2 and Bcl-xL proteins in HT-29 human colon cancer cells after CuO-NP exposure. Western blot assay showed downregulation of Bcl-2 and Bcl-xL protein expression after CuO-NP exposure. Our findings may aid in the understanding of the potential mechanisms responsible for induction of apoptosis owing to inhibition of Bcl-2 and Bcl-xL protein expression. Furthermore, the antibacterial activity assay showed that the synthesized CuO-NPs did not exert significant inhibitory effects against different gram-positive and gram-negative bacteria in vitro.

Design, synthesis and in vitro evaluation of anticancer and antibacterial potential of surface modified Tb(OH)3@SiO2 core–shell nanoparticles

In the current study, we modified the surface of Tb(OH)3 nanoparticles with a silica layer to enhance their solubility and biocompatibility. Transmission electron microscopy confirmed the improvements in these properties. Tb(OH)3@SiO2 core–shell nanoparticles (TS-CSNPs) exhibited a strong green emission peak upon irradiation with ultraviolet light, which originates from the electric-dipole transition 5D4 → 7F5 (543 nm) of the Tb3+ ion. In vitro anticancer and antimicrobial activities of the synthesized TS-CSNPs has been evaluated through their potential cytotoxicity and antibacterial activity. TS-CSNPs were shown to have more cytotoxicity against HT29 human colorectal cancer cells with a value of IC50 420.33 in an MTT assay. The alteration of the morphological features of HT29 cells was analysed using various concentrations of TS-CSNPs by inverted microscopy. Western blot analysis results of the apoptotic pathway showed that TS-CSNPs inhibited the growth of HT29 cancer cells through the induction of apoptosis, as evidenced by the down regulation of the expression of anti-apoptotic Bcl-2 and Bcl-xL gene products. Furthermore, the results of the in vitro hemolysis assay with human erythrocytes demonstrated the excellent blood biocompatibility of TS-CSNPs. Our silica coated TS-CSNPs exhibited a non-significant effect on the viability of both Gram negative and Gram positive bacterial strains up to 18 hours of exposure. These results highlight that modified TS-CSNPs can be functionalized to enhance the efficacy of cancer therapeutics due to the significant potential against cancerous cells and antibacterial activity.

Evaluation of in vitro cytotoxicity, biocompatibility, and changes in the expression of apoptosis regulatory proteins induced by cerium oxide nanocrystals

Abstract Cerium oxide nanocrystals (CeO2-NCs) exhibit superoxide dismutase and catalase mimetic activities. Based on these catalytic activities, CeO2-NCs have been suggested to have the potential to treat various diseases. The crystalline size of these materials is an important factor that influences the performance of CeO2-NCs. Previous reports have shown that several metal-based nanocrystals, including CeO2-NCs, can induce cytotoxicity in cancer cells. However, the underlying mechanisms have remained unclear. To characterize the anticancer activities of CeO2-NCs, several assays related to the mechanism of cytotoxicity and induction of apoptosis has been performed. Here, we have carried out a systematic study to characterize CeO2-NCs phase purity (X-ray diffraction), morphology (electron microscopy), and optical features (optical absorption, Raman scattering, and photoluminescence) to better establish their potential as anticancer drugs. Our study revealed anticancer effects of CeO2-NCs in HT29 and SW620 colorectal cancer cell lines with half-maximal inhibitory concentration (IC50) values of 2.26 and 121.18 μg ml–1, respectively. Reductions in cell viability indicated the cytotoxic potential of CeO2-NCs in HT29 cells based on inverted and florescence microscopy assessments. The mechanism of cytotoxicity confirmed by estimating possible changes in the expression levels of Bcl2, BclxL, Bax, PARP, cytochrome c, and β-actin (control) proteins in HT29 cells. Down-regulation of Bcl2 and BclxL and up-regulation of Bax, PARP, and cytochrome c proteins suggested the significant involvement of CeO2-NCs exposure in the induction of apoptosis. Furthermore, biocompatibility assay showed minimum effect of CeO2-NCs on human red blood cells.