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Recently, non-small cell lung cancer (NSCLC) has been the prime concern of cancer clinicians due to its high mortality rate worldwide. Cisplatin, a platinum derivative, has been used as a therapeutic option for treating metastatic NSCLC for several years. However, acquired, or intrinsic drug resistance to Cisplatin is the major obstacle to the successful treatment outcome of patients. Dysregulation of Nrf2 (nuclear factor erythroid 2-related factor 2) and EGFR (epidermal growth factor receptor) signaling have been associated with cellular proliferation, cancer initiation, progression and confer drug resistance to several therapeutic agents including Cisplatin in various cancers. To dissect the molecular mechanism of EGFR activation in resistant cells, we developed Cisplatin-resistant (CisR) human NSCLC cell lines (A549 and NCIH460) with increasing doses of Cisplatin treatment over a 3-month period. CisR cells demonstrated increased proliferative capacity, clonogenic survivability and drug efflux activity compared to the untreated parental (PT) cells. These resistant cells also showed higher levels of Nrf2 and EGFR expression. Here, we found that Nrf2 upregulates both basal and inducible expression of EGFR in these CisR cells at the transcriptional level. Moreover, genetic inhibition of Nrf2 with siRNA in CisR cells showed increased sensitivity towards the EGFR tyrosine kinase inhibitor (TKIs), AG1478. Our study, therefore suggests the use of Nrf2 inhibitors in combinatorial therapy with EGFR TKIs for the treatment of resistant NSCLC.
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We developed ZnO-assisted 1393 bioactive glass-based scaffold with suitable mechanical properties through foam replica technique and observed to be suitable for bone tissue engineering application. However, the developed scaffolds' ability to facilitate cellular infiltration and integration was further assessed through in vivo studies in suitable animal model. Herein, the pure 1393 bioactive glass (BG) and ZnO-assisted 1393 bioactive glass- (ZnBGs; 1, 2, 4 mol% ZnO substitution for SiO2 in pure BG is named as Z1BG, Z2BG, Z3BG, respectively) based scaffolds were prepared through sol-gel route, followed by foam replica techniques and characterized by a series of in vitro and some in vivo tests. Different cell lines like normal mouse embryonic cells (NIH/3T3), mouse bone marrow stromal cells (mBMSc), peripheral blood mononuclear cells, that is, lymphocytes and monocytes (PBMC) and U2OS (carcinogenic human osteosarcoma cells) were used in determination and comparative analysis of the biological compatibility of the BG and ZnBGs. Also, the alkaline phosphatase (ALP) activity, and osteogenic gene expression by primer-specific osteopontin (OPN), osteocalcin (OCN), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genes were performed to study osteogenic differentiability of the stromal cells in different BGs. Moreover, radiological and histopathological tests were performed in bone defect model of Wister rats to evaluate the in vivo bone regeneration and healing. Interestingly, these studies demonstrate augmented biological compatibility, and superior osteogenic differentiation in ZnBGs, in particular Z3BG than the pure BG in most cases.
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Neoplasias Óseas , Óxido de Zinc , Animales , Humanos , Ratas , Ratones , Osteogénesis , Leucocitos Mononucleares , Óxido de Zinc/farmacología , Dióxido de Silicio , Ratas Wistar , Vidrio , Diferenciación Celular , Andamios del TejidoRESUMEN
In clinical practice, major efforts are underway to identify appropriate drug combinations to boost anticancer activity while suppressing unwanted adverse effects. In this regard, we evaluated the efficacy of combination treatment with the widely used chemotherapeutic drug doxorubicin along with the TGFßRI inhibitor galunisertib (LY2157299) in aggressive B-cell non-Hodgkin lymphoma (B-NHL). The antiproliferative effects of these drugs as single agents or in combination against several B-NHL cell lines and the synergism of the drug combination were evaluated by calculating the combination index. To understand the putative molecular mechanism of drug synergism, the TGF-ß and stress signaling pathways were analyzed after combination treatment. An aggressive lymphoma model was used to evaluate the anticancer activity and post-therapeutic immune response of the drug combination in vivo. Galunisertib sensitized various B-NHL cells to doxorubicin and in combination synergistically increased apoptosis. The antitumor activity of the drug combinations involved upregulation of p-P38 MAPK and inhibition of the TGF-ß/Smad2/3 and PI3K/AKT signaling pathways. Combined drug treatment significantly reduced tumor growth and enhanced survival, indicating that the synergism between galunisertib and Dox observed in vitro was most likely retained in vivo. Based on the tumor-draining lymph node analysis, combination therapy results in better prognosis, including disappearance of disease-exacerbating regulatory T cells and prevention of CD8+ T-cell exhaustion by downregulating MDSCs. Galunisertib synergistically potentiates the doxorubicin-mediated antitumor effect without aggravating the toxic effects and the ability to kickstart the immune system, supporting the clinical relevance of targeting TGF-ßRI in combination with doxorubicin against lymphoma.
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Linfoma , Neoplasias , Humanos , Fosfatidilinositol 3-Quinasas/farmacología , Doxorrubicina/farmacología , Doxorrubicina/uso terapéutico , Neoplasias/tratamiento farmacológico , Factor de Crecimiento Transformador beta , Sistema Inmunológico , Sinergismo Farmacológico , Línea Celular Tumoral , ApoptosisRESUMEN
BACKGROUND AIMS: The stimulatory natural killer-dendritic cell axis in the tumor microenvironment could play a critical role in stimulating cytotoxic T cells and driving immune responses against cancer. METHODS: We established a novel treatment protocol by adroitly combining chemotherapy with doxorubicin and immunotherapy with dendritic cells and natural killer cells against a highly aggressive and malignant lymphoma called Dalton's lymphoma. RESULTS: Our data suggest that binary application of adoptive cell therapy and chemotherapy nearly cures (95%) early-stage experimental lymphoma. In the case of mid-stage cancer, the success rate was significantly lower but still impressive (75%). Our results demonstrated that the application of combination therapy in early-stage cancer significantly reduced the tumor volume and extended the lifespan of the experimental animal in addition to reinvigorating the immune system, including restoring the effector functions of dendritic cells and natural killer cells. The novel protocol limits the metastasis of tumor cells in vascularized organs and rearms the adaptive immune response mediated by dendritic cells and CD4+ and CD8+ T cells. CONCLUSIONS: Combination therapy in the early stage alters the cytokine profile, increases interferon-γ and tumor necrosis factor-α in the serum of treated animals and downregulates programmed cell death protein 1 expression in CD8+ T cells. Thus, cooperative and cognitive interactions between dendritic cells and natural killer cells in addition to therapy with doxorubicin promote the immune response and tumoricidal activities against lymphoma.
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Linfoma , Receptor de Muerte Celular Programada 1 , Animales , Citocinas , Linfocitos T CD8-positivos , Linfoma/terapia , Células Asesinas Naturales , Doxorrubicina/farmacología , Doxorrubicina/uso terapéutico , Células Dendríticas , Factores de Transcripción Forkhead , Microambiente TumoralRESUMEN
T-cell exhaustion plays a pivotal role in the resistance of microsatellite-stable colorectal cancer (CRC) to immunotherapy. Identifying and targeting T-cell exhaustion-activating mechanisms is a promising strategy to augment the effects of immunotherapy. Here, we found that thymidine phosphorylase (TYMP) plays a decisive role in inducing systemic T-cell exhaustion and abrogating the efficacy of dendritic cell (DC) therapy in a CRC model. Targeting TYMP with tipiracil hydrochloride (TPI) induces immunological cell death (ICD). The combined effects of TPI and imiquimod-activated DCs turn CT26 tumors into immunologically 'hot' tumors by inducing ICD in vivo. High-dimensional cytometry analysis revealed T-cell and IFN-γ dependency on the therapeutic outcome. In addition, chemoimmunotherapy converts intratumoral Treg cells into Th1 effector cells and eliminates tumor-associated macrophages, resulting in higher cytotoxic T lymphocyte infiltration and activation. This effect is also associated with the downregulation of PD-L1 expression in tumors, leading to the prevention of T-cell exhaustion. Thus, cooperative and cognitive interactions between dendritic cells and immunogenic cell death induced by therapy with TPI promote the immune response and tumoricidal activities against microsatellite stable colorectal cancer. Our results support TYMP targeting to improve the effects of DC immunotherapy and outcomes in CRC.
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Neoplasias Colorrectales , Timidina Fosforilasa , Células Dendríticas , Humanos , Factores Inmunológicos , Inmunoterapia/métodosRESUMEN
The presence of ERG gene fusion; from developing prostatic intraepithelial neoplasia (PIN) lesions to hormone resistant high grade prostate cancer (PCa) dictates disease progression, altered androgen metabolism, proliferation and metastasis1-3. ERG driven transcriptional landscape may provide pro-tumorigenic cues in overcoming various strains like hypoxia, nutrient deprivation, inflammation and oxidative stress. However, insights on the androgen independent regulation and function of ERG during stress are limited. Here, we identify PGC1α as a coactivator of ERG fusion under various metabolic stress. Deacetylase SIRT1 is necessary for PGC1α-ERG interaction and function. We reveal that ERG drives the expression of antioxidant genes; SOD1 and TXN, benefitting PCa growth. We observe increased expression of these antioxidant genes in patients with high ERG expression correlates with poor survival. Inhibition of PGC1α-ERG axis driven transcriptional program results in apoptosis and reduction in PCa xenografts. Here we report a function of ERG under metabolic stress which warrants further studies as a therapeutic target for ERG fusion positive PCa.
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Antioxidantes , Coactivador 1-alfa del Receptor Activado por Proliferadores de Peroxisomas gamma , Neoplasias de la Próstata , Andrógenos , Antioxidantes/farmacología , Fusión Génica , Humanos , Masculino , Coactivador 1-alfa del Receptor Activado por Proliferadores de Peroxisomas gamma/genética , Coactivador 1-alfa del Receptor Activado por Proliferadores de Peroxisomas gamma/metabolismo , Neoplasias de la Próstata/patología , Estrés Fisiológico , Regulador Transcripcional ERG/genética , Regulador Transcripcional ERG/metabolismoRESUMEN
Integrative medicine practices, such as Ayurveda, are popular in India and many South Asian countries, yet basic research to investigate the concepts, procedures, and medical benefits of ayurvedic products has received little attention and is not fully understood. Here, we report a functional nanodiamond-based traditional Ayurvedic herbomineral formulation, Heerak Bhasma (Ayu_ND), for the treatment of solid tumors called Dalton's lymphoma generated in CD1 mice. Ayu_ND-mediated immunostimulation significantly reduces tumor cell proliferation and induces apoptosis aided by the active participation of dendritic cells. Immunomodulatory Ayu_ND treatment is highly immunostimulatory and drives dendritic cells to produce TNF-α. Treatment with Ayu_ND significantly reduces the tumor volume, inhibits metastasis in distant vascularized organs, and increases the life span of tumor-bearing animals compared with untreated littermates. These events were associated with elevated serum levels of the protective cytokines IFN-γ and TNF-α and downregulated the disease, exacerbating TGF-ß. Ayu_ND-mediated therapeutic success was also accompanied by the depletion of regulatory T cells and enhanced vaccine-induced T-cell immunity, guided by the restoration of the memory CD8+ T-cell pool and prevention of PD-1-mediated T cell exhaustion. The results provide a basis for further evaluation of ayurvedic formulations and drug efficacy in treating cancers.
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Despite several extensive and exhaustive efforts, search for potential therapy against leishmaniasis has not made much progress. In the present work, we have employed mining strategy to screen Leishmania donovani proteome for identification of promising vaccine candidate. We have screened 21 potential antigenic proteins from 7960 total protein of L. donovani, based on the presence of signal peptide, GPI anchor, antigenicity prediction and substractive proteomic approach. Secondly, we have also performed comprehensive immunogenic epitope prediction from the screened 21 proteins, using IEDB-AR tools. Out of the 21 antigenic proteins, we obtained 11 immunogenic epitopes from 9 proteins. The final results revealed that four predicted epitopes namely; YPAFAALVF, VAVAATVAY, AAAPTEAAL and MYPLVAVVF, have significantly better binding potential with respective alleles and could elicits immune responses. Docking analysis using PATCHDOCK server and molecular dynamic simulation using GROMACS revealed the potential of the sequences as immunogenic epitopes. In silico studies also suggested that the epitopes occupied almost same binding cleft with the respective alleles, when compared with the reference peptides. It is also suggested from the molecular dynamic simulation data that the peptides were intact in the pocket for longer periods of time. Our study was designed to select MHC class I restricted epitopes for the activation of CD8 T cells using immunoinformatics for the prediction of probable vaccine candidate against L. donovani parasites. Communicated by Ramaswamy H. Sarma.
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Leishmania donovani , Vacunas , Proteoma , Proteómica , Epítopos de Linfocito T , Biología Computacional/métodos , Péptidos , Desarrollo de Vacunas , Simulación del Acoplamiento Molecular , Epítopos de Linfocito B , Vacunas de SubunidadRESUMEN
We fabricated bilirubin-bovine serum albumin (BR-BSA) nanocomplexes as candidates for the delivery of 5-fluoro-2-deoxyuridine (5FUdr) against experimental murine lymphoma. BR was attached to 5FUdr via acid-labile ester bonds mimicking small-molecule drug conjugates. The construct was self-assembled with BSA through strong noncovalent interactions with high drug occupancy in the core and labeled with folic acid (FA) to target cancer cells. The BR-5FUdr-BSA-FA nanoconstruct exhibits excellent biocompatibility, prevents nephrotoxicity, and is tolerated by red blood cells and mononuclear cells. The construct also showed increased accumulation in lymph nodes and tumor cells. BR-5FUdr-BSA-FA caused prolonged growth inhibition and apoptosis, enhanced mitochondrial reactive oxygen species generation, and minimized the viability of parental and doxorubicin-resistant Dalton's lymphoma cells. Treatment of tumor-bearing mice with BR-5FUdr-BSA-FA significantly increased the life span of the animals, improved their histopathological parameters, and downregulated PD-1 expression, suggesting the potential of the construct for 5FUdr delivery to treat lymphoma.
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Desoxiuridina/análogos & derivados , Portadores de Fármacos/química , Linfoma/tratamiento farmacológico , Receptor de Muerte Celular Programada 1/antagonistas & inhibidores , Animales , Apoptosis/efectos de los fármacos , Bilirrubina/química , Materiales Biomiméticos/química , Linfocitos T CD8-positivos/efectos de los fármacos , Linfocitos T CD8-positivos/metabolismo , Línea Celular Tumoral , Desoxiuridina/administración & dosificación , Desoxiuridina/farmacocinética , Modelos Animales de Enfermedad , Regulación hacia Abajo/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Linfoma/patología , Ratones , Receptor de Muerte Celular Programada 1/metabolismo , Albúmina Sérica Bovina/químicaRESUMEN
In the present study, a magnetic nanocomposite (magnetite Fe3O4 and hematite Fe2O3) has been successfully synthesized by the sol-gel method and coated with polyvinyl alcohol (PVA) followed by conjugation of anti-diabetic drug metformin. Detailed structural and microstructural characterization of the nanocomposite (NP) and drug conjugated nanocomposite (NP-DC) are analyzed by the Rietveld refinement of respective XRD patterns, FTIR analysis, UV-Vis spectroscopy, SEM and TEM results. SEM and TEM image analyses reveal the spherical morphology and average size of NP, PVA coated nanoparticles (NP-PVA) and NP-DC samples, indicating a suitable size to be a nanocarrier. The biocompatibility of NP and NP-DC was carried out in NIH/3T3 and J774A. 1 cells. The enhanced activity of the drug, when conjugated with nanocomposite, is confirmed after the treatment of both the pure drug and NP-DC sample on the 18 h fasted normoglycemic and hyperglycemic mice. The blood glucose level of the mice is effectively decreased with the same concentration of the pure drug and NP-DC sample. It proves the increased activity of the NP-DC sample, as only 5 wt% drug is present that shows the same efficiency as the pure drug. This study suggests excellent biocompatibility and cytocompatibility of NP and NP-DC besides the critical property as a hypoglycemic agent. It is the first time approach of conjugating metformin with a magnetic nanocomposite for a significant increment of its hypoglycemic activity, which is very important to reduce the side effect of metformin for its prolonged use.
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Nanocompuestos , Preparaciones Farmacéuticas , Animales , Hipoglucemiantes/farmacología , Fenómenos Magnéticos , Magnetismo , RatonesRESUMEN
Enhanced drug localization at the tumor sites with minimal toxicity was demonstrated using dendrimer-conjugated temozolomide for treating experimental lymphoma, developed as a solid tumor. Herein, we have constructed a polyamidoamine (PAMAM) dendrimer conjugated with temozolomide to enhance the stability of the active drug metabolites, derived from the prodrug temozolomide. Our results suggest that the active drug (5-(3-methyltriazen-1-yl)imidazole-4-carboxamide) (MTIC) (derived from temozolomide) showed stable and sustained release from the dendrimer-temozolomide conjugate, suggesting the suitability of the construct for therapy. Besides growth inhibition and direct killing, the dendrimer-temozolomide construct induced extensive apoptosis not only in parental Dalton lymphoma tumor cells but also in the doxorubicin-resistant form of the tumor cells. Dendrimer-temozolomide conjugation significantly reduced the solid tumor growth and increased the lifespan with better prognosis, including improved histopathology of the treated mice, while untreated littermates developed extensive metastasis and succumbed to death.
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Antineoplásicos Alquilantes/farmacología , Materiales Biocompatibles/farmacología , Dendrímeros/farmacología , Desarrollo de Medicamentos , Linfoma/tratamiento farmacológico , Temozolomida/farmacología , Animales , Antineoplásicos Alquilantes/síntesis química , Antineoplásicos Alquilantes/química , Apoptosis/efectos de los fármacos , Materiales Biocompatibles/síntesis química , Materiales Biocompatibles/química , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Dendrímeros/química , Liberación de Fármacos , Ensayos de Selección de Medicamentos Antitumorales , Femenino , Humanos , Linfoma/patología , Ensayo de Materiales , Ratones , Estructura Molecular , Neoplasias Experimentales/tratamiento farmacológico , Neoplasias Experimentales/patología , Tamaño de la Partícula , Temozolomida/químicaRESUMEN
Galunisertib (LY2157299) is a selective ATP-mimetic inhibitor of TGF-ß receptor-I activation, currently under clinical trial in a variety of cancers. We have tested the combined effects of galunisertib- and interleukin-15-activated dendritic cells in an aggressive and highly metastatic murine lymphoma. Based on the tumor-draining lymph node architecture, and its histology, the combination therapy results in better prognosis, including disappearance of the disease-exacerbating regulatory T cells. Our data suggest that galunisertib significantly enhances the success of immunotherapy with IL-15-activated dendritic cells by limiting the regulatory T cells generation with consequent downregulation of regulatory T cells in the tumor-draining lymph nodes and vascularized organ like spleen. This is also associated with consistent loss p-SMAD2 and downregulation of Neuropilin-1, leading to better prognosis and positive outcome. These results connect the role of combined therapy with the consequent elimination of disease-exacerbating T regulatory cells in a metastatic murine lymphoma.
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Peroxidases are a heterogeneous family of enzymes that have diverse biological functions. Ascorbate peroxidase is a redox enzyme that is reduced by trypanothione, which plays a central role in the redox defense system of Leishmania In view of developing new and novel therapeutics, we performed in silico studies in order to search for a ligand library and identify new drug candidates and their physiological roles against promastigotes and intracellular amastigotes of Leishmania donovani Our results demonstrated that the selected inhibitor ZINC96021026 has significant antileishmanial effect and effectively killed both free and intracellular forms of the parasite. ZINC96021026 was found to be identical to ML-240, a selective inhibitor of valosin-containing protein (VCP), or p97, a member of the AAA-ATPase protein family which was derived from the scaffold of N2,N4-dibenzylquinazoline-2,4-diamine (DBeQ), targeting the D2-ATPase domain of the enzyme. ZINC96021026 (ML-240) thus has a broad range of cellular functions, thought to be derived from its ability to unfold proteins or disassemble protein complexes, besides inhibiting the ascorbate peroxidase activity. ML-240 may inhibit the parasite's ascorbate peroxidase, leading to extensive apoptosis and inducing generation of reactive oxygen species. Taken together, our results demonstrated that ML-240 could be an attractive therapeutic option for treatment against leishmaniasis.
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Antiprotozoarios , Ascorbato Peroxidasas/antagonistas & inhibidores , Leishmania donovani , Antiprotozoarios/farmacología , Simulación por Computador , Leishmania donovani/efectos de los fármacosRESUMEN
A new amide-imine conjugate, 2-hydroxybenzoic acid-(2-hydroxybenzylidene)-hydrazide (L1), is employed to prepare a single crystal X-ray structurally characterized poly-nuclear Cu(ii) complex (M1). M1 selectively and spatially interacts with cytochrome C (Cyt C) to allow fluorescence imaging of intracellular translocation events in living cells. Thus, direct visualization of a Cyt C translocation event during an apoptotic process is achieved for the first time. The binding constant and LOD are 7.52 × 104 M-1 and 34.0 nM, respectively.
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Complejos de Coordinación/química , Cobre/química , Citocromos c/metabolismo , Mitocondrias/metabolismo , Animales , Apoptosis/efectos de los fármacos , Línea Celular , Complejos de Coordinación/farmacología , Citocromos c/análisis , Humanos , Hidrazinas/química , Ratones , Microscopía Fluorescente , Mitocondrias/efectos de los fármacos , EspectrofotometríaRESUMEN
A triple stimuli-responsive drug delivery platform involving doxorubicin, 5-fluoro-2-deoxy uridine and folic acid was fabricated on mesoporous silica nanoparticles for targeting delivery against a highly aggressive murine lymphoma called Dalton's lymphoma. Fabrication of the unique construct by amalgamating active and passive targeting mechanisms offers a novel hyper-chimeric platform for a stimuli-responsive drug delivery system. The novel construct enables efficient and precise delivery of the precious cargo to the tumor sites. Active targeting by folic acid directs the doxorubicin and 5-fluoro-2-deoxy uridine in the close proximities of the tumor cells, causing efficient killing and significant growth inhibition. Isobologram models, zero interaction potency dose-response surface plots and matrices were generated to evaluate the combination synergism of the two drugs. Therapy with the dual drug-bearing construct in mice with established tumors significantly reduced the tumor load and enhanced the survival of the animals compared with the untreated control. Therapy with the dual delivery system also augmented the innate and adaptive immune defense mechanisms of the treated animals. CD8+ T cells, natural killer cells and the dendritic cells from the treated group following successful therapy with the novel construct showed enhanced cytotoxicity and growth inhibitory capacities against DL tumor cells.
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Antineoplásicos/farmacología , Desoxiuridina/análogos & derivados , Doxorrubicina/farmacología , Linfoma/tratamiento farmacológico , Animales , Antineoplásicos/química , Apoptosis/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Cisteína/química , Desoxiuridina/química , Desoxiuridina/farmacología , Disulfuros/química , Doxorrubicina/química , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Linfoma/patología , Ratones , Nanopartículas/química , Neoplasias Experimentales/tratamiento farmacológico , Neoplasias Experimentales/patología , Tamaño de la Partícula , Porosidad , Dióxido de Silicio/química , Propiedades de SuperficieRESUMEN
Cooperative and cognitive interaction between the dendritic cells and natural killer cells was investigated for demonstrating the anti-tumor activity against an aggressive murine lymphoma, treated with doxorubicin. Crosstalk between the dendritic cells and the natural killer cells significantly reduced the proliferation of Dalton's lymphoma cells in a dose dependent manner. Treatment of Dalton's lymphoma cells with doxorubicin in vitro enhances the effects of crosstalk against the target cells. This crosstalk between the cells was regulated via stimulation with recombinant interleukin-15, and release of TNF-α which is critically important for the tumoricidal effects. Dendritic cells and the natural killer cells crosstalk activate both the cells and upregulate the expression of CD40, CD69 and CD86 on the dendritic cells. These findings provided new insight regarding these interactions and define a mechanism by which cellular immune response promotes tumoricidal activity against lymphoma in therapeutic setting.
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Células Dendríticas/inmunología , Interleucina-15/administración & dosificación , Células Asesinas Naturales/inmunología , Linfoma/inmunología , Factor de Necrosis Tumoral alfa/metabolismo , Animales , Comunicación Celular/efectos de los fármacos , Comunicación Celular/inmunología , Línea Celular Tumoral/trasplante , Proliferación Celular/efectos de los fármacos , Células Dendríticas/metabolismo , Modelos Animales de Enfermedad , Doxorrubicina/administración & dosificación , Femenino , Regulación Neoplásica de la Expresión Génica/inmunología , Humanos , Inmunidad Celular/efectos de los fármacos , Interferón gamma/inmunología , Interferón gamma/metabolismo , Interleucina-15/inmunología , Linfoma/tratamiento farmacológico , Linfoma/patología , Ratones , Cultivo Primario de Células , Proteínas Recombinantes/administración & dosificación , Proteínas Recombinantes/inmunología , Regulación hacia ArribaRESUMEN
A novel ABA-type polyethylene glycol (PEG)-b-polyketal (PK)-b-PEG block copolymer was synthesized via click reactions between the monoazido-monomethoxy-PEG and dialkyne terminated aliphatic polyketal with no carboxylic/amide linkages. Formation of the novel block copolymer was confirmed by 1H NMR, GPC, TGA, and DSC studies. The formed copolymer has shown faster degradation at acidic pH. Self-assembly of this block copolymer (average size 6.2 nm) was assessed by fluorescence study using pyrene as a probe. Doxorubicin loaded block copolymeric micelles (69.9 nm) have shown pH dependent elevated drug release at pH 6.4, indicating its potential as a pH responsive nano-carrier for anticancer therapy. The nano-sized copolymer demonstrated tumoricidal activities against the lymphoma of murine and human origin with significant levels of growth inhibition and apoptosis. Therapy with doxorubicin loaded copolymer reduced the tumor size and augmented the life span of the tumor bearing animals with improved histopathological parameters, compared with the untreated control.
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Materiales Biocompatibles/química , Doxorrubicina/química , Polietilenglicoles/química , Animales , Portadores de Fármacos/química , Concentración de Iones de Hidrógeno , Ratones , Micelas , Polietilenos/química , Polímeros/químicaRESUMEN
The present work deals with the identification and characterization of a novel inhibitor Z220582104, specific to pyruvate phosphate dikinase, for leishmanicidal activities against free promastigotes and intracellular amastigotes. We have used structure-based drug designing approaches and performed homology modelling, virtual screening and molecular dynamics studies. Primary mouse macrophages and macrophage cell line J774A1 were infected with promastigotes of Leishmania donovani. Both promastigotes and infected macrophages were subjected to treatment with the varying concentrations of Z220582104 or miltefosine for assessment of leishmanicidal activity. The novel inhibitor Z220582104 demonstrated growth inhibitory potential and reduced the viability of the free promastigotes in a concentration- and time-dependent manner. Z220582104 was also effective against the intracellular form of the parasites and reduced the number of amastigotes in macrophages and also lowered the parasite index, compared with the untreated infected macrophages. Although less effective compared with the miltefosine, Z220582104 is well tolerated by the dividing cells and normal human lymphocytes and monocytes with no adverse effects on the growth kinetics or viability. Our in silico and in vitro studies suggested that Leishmania donovani pyruvate phosphate dikinase could be a potential new drug target.
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Antiparasitarios/farmacología , Leishmania donovani/efectos de los fármacos , Leishmania donovani/crecimiento & desarrollo , Macrófagos/parasitología , Piruvato Ortofosfato Diquinasa/antagonistas & inhibidores , Animales , Células Cultivadas , Diseño de Fármacos , Humanos , Leishmania donovani/aislamiento & purificación , Ratones , Simulación de Dinámica Molecular , Fosforilcolina/análogos & derivados , Fosforilcolina/farmacologíaRESUMEN
Single crystal X-ray structurally characterized benzimidazole-naphthalene hybrid (NABI) functions as a unique dual analyte sensor that can detect Zn2+ cation and N3- anion independently. The NABI forms chelate with Zn2+ to inhibit internal charge transfer (ICT) and CHN isomerisation resulting chelation enhanced fluorescence (CHEF). On the other hand, the sensing of N3- is based on formation of supramolecular H-bonded rigid assembly. The association constant of NABI for Zn2+ and N3- ions are 19â¯×â¯104â¯M-1 and 11â¯×â¯102â¯M-1, respectively. Corresponding limit of detections (LOD) are 6.85â¯×â¯10-8 and 1.82â¯×â¯10-7â¯M, respectively. NABI efficiently detects intracellular Zn2+ and N3- ions with no cytotoxicity on J774A.1cells under fluorescence microscope. DFT studies unlock underlying spectroscopic properties of free NABI and Zn2+/N3- bound forms.
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Three new amide-imine conjugates, namely [(E)-2-hydroxy-N'-((2-hydroxynaphthalen-1-yl)methylene)benzohydrazide] (SALNP), [(E)-N'-(4-(diethylamino)-2-hydroxybenzylidene)-2-hydroxybenzohydrazide] (SALSD), and [(E)-N'-(3-ethoxy-4-hydroxybenzylidene)-2-hydroxybenzohydrazide] (SALVN), derived by reacting 2-hydroxybenzohydrazide (SAL) with three different aldehyde, 2-hydroxynapthaldehyde, 4-(diethylamino)-2-hydroxybenzaldehyde, and 3-ethoxy-4-hydroxybenzaldehyde, respectively. Three mononuclear oxovanadium(V) and two µoxo-bridged dinuclear molybdenum(VI) complexes have been synthesized using SALNP and SALSD. Besides, SALVN is used to prepare oxovanadium(V) and dioxomolybdenum(VI) complexes. All five metal complexes along with three amide-imine conjugates are characterized by single crystal XRD analysis. Some of them have been explored as catalyst for oxidation of alkyl benzene and styrene. Antitumor activities of the metal complexes along with ligands have been studied on Dalton lymphoma (DL) and 2PK3 murine lymphoma cells.