RESUMEN
The high heterogeneity of neuroblastoma (NB) is currently the main challenge in clinical treatment, impeding the complete eradication of the tumor through monotherapy alone. In this study, we propose a combination strategy using a targeted nano co-delivery system (ADRF@Ag2Se) comprising phototheranostic agents, differentiation inducers and chemotherapy drugs for sequential therapy of NB. Upon intravenous injection, ADRF@Ag2Se demonstrates effective tumor targeting by the specific binding of AF7P to MMP14, which is overexpressed on the surface of NB cells. Subsequent implementation of local photothermal therapy (PTT) leverages the robust photothermal conversion capabilities of the amphiphilic photothermal reagent PF. This is followed by the temperature-triggered release of differentiation-inducing agent 13-cis-retinoic acid and chemo-drug doxorubicin to synergistically eliminate the residual lesions. This nanotherapeutic strategy facilitatesin vivotargeted delivery and PTT under the supervision of NIR-II fluorescence, and it also enhances the chemotherapeutic response through differentiation induction of poorly differentiated cancer cells. In the NB tumor model, this co-delivery strategy effectively inhibited tumor growth and significantly prolonged the survival of the mice.
Asunto(s)
Doxorrubicina , Neuroblastoma , Neuroblastoma/tratamiento farmacológico , Neuroblastoma/patología , Neuroblastoma/metabolismo , Animales , Ratones , Línea Celular Tumoral , Humanos , Doxorrubicina/farmacología , Doxorrubicina/química , Doxorrubicina/administración & dosificación , Terapia Fototérmica , Sistemas de Liberación de Medicamentos , Antineoplásicos/química , Antineoplásicos/farmacología , Antineoplásicos/administración & dosificación , Ratones Desnudos , Tretinoina/química , Tretinoina/farmacología , Tretinoina/administración & dosificación , Nanopartículas/química , Diferenciación Celular/efectos de los fármacos , Metaloproteinasa 14 de la Matriz/metabolismoRESUMEN
As a prescription drug, retinoic acid is listed as a banned cosmetic additive in the EU and China regulations. Currently, spectrophotometric methods, including thin-layer chromatography (TLC), high-performance liquid chromatography (HPLC), and HPLC-MS/MS, are commonly used for the determination of retinoic acid. As these conventional methods require complex pretreatment and are time-consuming, chemical derivatization combined with paper spray ionization mass spectrometry was developed for the fast detection of retinoic acid in cosmetics. N,N-dimethylpiperazine iodide (DMPI) was utilized as a derivatization reagent. Carboxylic acid in retinoic acid was derivatized to carry a positive charge and was subjected to mass spectrometry analysis. Results showed that compared with non-derivatized compounds, the detection limit was increased by about 50 times. The linearity in the range of 0.005-1 µg·mL-1 was good. The limit of detection (LOD) was 0.0013 µg·mL-1, and the limit of quantification (LOQ) was 0.0043 µg·mL-1. The recoveries of spiked samples were in the range of 95-105%, and the RSDs were below 5%. Derivatization and paper spray ionization MS render a quick, sensitive, and accurate method for the detection of retinoic acid in a complex matrix.
Asunto(s)
Cosméticos , Tretinoina , Tretinoina/análisis , Tretinoina/química , Cosméticos/química , Cosméticos/análisis , Límite de Detección , Papel , Cromatografía Líquida de Alta Presión/métodos , Espectrometría de Masas en Tándem/métodos , Espectrometría de Masas/métodosRESUMEN
Nonalcoholic fatty liver disease (NAFLD) is characterized by excessive lipid accumulation, steatosis and fibrosis. Sympathetic nerves play a critical role in maintaining hepatic lipid homeostasis and regulating fibrotic progression through adrenergic receptors expressed by hepatocytes and hepatic stellate cells; however, the use of sympathetic nerve-focused strategies for the treatment of NAFLD is still in the infancy. Herein, a biomimetic nanoplatform with ROS-responsive and ROS-scavenging properties was developed for the codelivery of retinoic acid (RA) and the adrenoceptor antagonist labetalol (LA). The nanoplatform exhibited improved accumulation and sufficient drug release in the fibrotic liver, thereby achieving precise codelivery of drugs. Integration of adrenergic blockade effectively interrupted the vicious cycle of sympathetic nerves with hepatic stellate cells (HSCs) and hepatocytes, which not only combined with RA to restore HSCs to a quiescent state but also helped to reduce hepatic lipid accumulation. We demonstrated the excellent ability of the biomimetic nanoplatform to ameliorate liver inflammation, fibrosis and steatosis. Our work highlights the tremendous potential of a sympathetic nerve-focused strategy for the management of NAFLD and provides a promising nanoplatform for the treatment of NAFLD.
Asunto(s)
Células Estrelladas Hepáticas , Enfermedad del Hígado Graso no Alcohólico , Enfermedad del Hígado Graso no Alcohólico/tratamiento farmacológico , Enfermedad del Hígado Graso no Alcohólico/metabolismo , Animales , Ratones , Células Estrelladas Hepáticas/efectos de los fármacos , Células Estrelladas Hepáticas/metabolismo , Hepatocitos/efectos de los fármacos , Hepatocitos/metabolismo , Ratones Endogámicos C57BL , Tretinoina/farmacología , Tretinoina/química , Tretinoina/uso terapéutico , Masculino , Receptores Adrenérgicos/metabolismo , Humanos , Biomimética/métodos , Materiales Biomiméticos/química , Materiales Biomiméticos/farmacología , Hígado/efectos de los fármacos , Hígado/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Nanopartículas/químicaRESUMEN
The molecular activation mechanism of the nuclear retinoid X receptors (RXRs) crucially involves ligand-induced corepressor release and coactivator recruitment which mediate transcriptional repression or activation. The ability of RXR to bind diverse coactivators suggests that a coregulator-selective modulation by ligands may open an avenue to tissue- or gene-selective RXR activation. Here, we identified strong induction of peroxisome proliferator-activated receptor γ coactivator 1α (PGC1α) binding to RXR by a synthetic agonist but not by the endogenous ligand 9-cis retinoic acid. Structure-guided diversification of this lead resulted in a set of three structurally related RXR agonists with different ability to promote PGC1α recruitment in cell-free and cellular context. These results demonstrate that selective modulation of coregulator recruitment to RXR can be achieved with molecular glues and potentially open new therapeutic opportunities by targeting the ligand-induced RXR-PGC1α interaction.
Asunto(s)
Coactivador 1-alfa del Receptor Activado por Proliferadores de Peroxisomas gamma , Receptores X Retinoide , Coactivador 1-alfa del Receptor Activado por Proliferadores de Peroxisomas gamma/metabolismo , Humanos , Receptores X Retinoide/agonistas , Receptores X Retinoide/metabolismo , Ligandos , Tretinoina/farmacología , Tretinoina/química , Tretinoina/metabolismo , Relación Estructura-Actividad , Alitretinoína/farmacología , Alitretinoína/química , Alitretinoína/metabolismo , Unión Proteica , Células HEK293RESUMEN
BACKGROUND: Bone tissue engineering endows alternates to support bone defects/injuries that are circumscribed to undergo orchestrated process of remodeling on its own. In this regard, hydrogels have emerged as a promising platform that can confront irregular defects and encourage in situ bone repair. METHODS: In this study, we aimed to develop a new approach for bone tissue regeneration by developing an alginate based composite hydrogel incorporating selenium doped biphasic calcium phosphate nanoparticles, and retinoic acid. The fabricated hydrogel was physiochemically evaluated for morphological, bonding, and mechanical behavior. Additionally, the biological response of the fabricated hydrogel was evaluated on MC3T3-E1 pre-osteoblast cells. RESULTS: The developed composite hydrogel confers excellent biocompatibility, and osteoconductivity owing to the presence of alginate, and biphasic calcium phosphate, while selenium presents pro osteogenic, antioxidative, and immunomodulatory properties. The hydrogels exhibited highly porous microstructure, superior mechanical attributes, with enhanced calcification, and biomineralization abilities in vitro. SIGNIFICANCE: By combining the osteoconductive properties of biphasic calcium phosphate with multifaceted benefits of selenium and retinoic acid, the fabricated composite hydrogel offers a potential transformation in the landscape of bone defect treatment. This strategy could direct a versatile and effective approach to tackle complex bone injuries/defects and present potential for clinical translation.
Asunto(s)
Alginatos , Regeneración Ósea , Hidrogeles , Selenio , Tretinoina , Regeneración Ósea/efectos de los fármacos , Hidrogeles/química , Hidrogeles/farmacología , Alginatos/química , Tretinoina/farmacología , Tretinoina/química , Animales , Ratones , Selenio/química , Selenio/farmacología , Osteogénesis/efectos de los fármacos , Hidroxiapatitas/química , Hidroxiapatitas/farmacología , Calcificación Fisiológica/efectos de los fármacos , Inmunomodulación/efectos de los fármacos , Línea Celular , Osteoblastos/efectos de los fármacos , Ingeniería de Tejidos/métodos , Nanopartículas/química , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Andamios del Tejido/químicaRESUMEN
Increasing the penetration and accumulation of antitumor drugs at the tumor site are crucial in chemotherapy. Smaller drug-loaded nanoparticles (NPs) typically exhibit increased tumor penetration and more effective permeation through the nuclear membrane, whereas larger drug-loaded NPs show extended retention at the tumor site. In addition, cancer stem cells (CSCs) have unlimited proliferative potential and are crucial for the onset, progression, and metastasis of cancer. Therefore, a drug-loaded amphiphilic peptide, DDP- and ATRA-loaded Pep1 (DA/Pep1), is designed that self-assembles into spherical NPs upon the encapsulation of cis-diamminedichloroplatinum (DDP) and all-trans retinoic acid (ATRA). In an acidic environment, DA/Pep1 transforms into aggregates containing sheet-like structures, which significantly increases drug accumulation at the tumor site, thereby increasing antitumor effects and inhibiting metastasis. Moreover, although DDP treatment can increase the number of CSCs present, ATRA can induce the differentiation of CSCs in breast cancer to increase the therapeutic effect of DDP. In conclusion, this peptide nanodelivery system that transforms in response to the acidic tumor microenvironment is an extremely promising nanoplatform that suggests a new idea for the combined treatment of tumors.
Asunto(s)
Neoplasias de la Mama , Nanopartículas , Péptidos , Tretinoina , Femenino , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/patología , Neoplasias de la Mama/metabolismo , Concentración de Iones de Hidrógeno , Péptidos/química , Humanos , Animales , Nanopartículas/química , Tretinoina/química , Tretinoina/farmacología , Tretinoina/farmacocinética , Portadores de Fármacos/química , Línea Celular Tumoral , Ratones , Células Madre Neoplásicas/efectos de los fármacos , Células Madre Neoplásicas/metabolismo , Cisplatino/química , Cisplatino/farmacología , Cisplatino/farmacocinética , Antineoplásicos/química , Antineoplásicos/farmacología , Ratones Endogámicos BALB CRESUMEN
Given the worldwide problem posed by enteric pathogens, the discovery of safe and efficient intestinal adjuvants combined with novel antigen delivery techniques is essential to the design of mucosal vaccines. In this work, we designed poly (lactic-co-glycolic acid) (PLGA)-based nanoparticles (NPs) to codeliver all-trans retinoic acid (atRA), novel antigens, and CpG. To address the insolubility of the intestinal adjuvant atRA, we utilized PLGA to encapsulate atRA and form a "nanocapsid" with polydopamine. By leveraging polydopamine, we adsorbed the water-soluble antigens and the TLR9 agonist CpG onto the NPs' surface, resulting in the pathogen-mimicking PLPCa NPs. In this study, the novel fusion protein (HBf), consisting of the Mycobacterium avium subspecies paratuberculosis antigens HBHA, Ag85B, and Bfra, was coloaded onto the NPs. In vitro, PLPCa NPs were shown to promote the activation and maturation of bone marrow-derived dendritic cells. Additionally, we found that PLPCa NPs created an immune-rich microenvironment at the injection site following intramuscular administration. From the results, the PLPCa NPs induced strong IgA levels in the gut in addition to enhancing powerful systemic immune responses. Consequently, significant declines in the bacterial burden and inflammatory score were noted in PLPCa NPs-treated mice. In summary, PLPCa can serve as a novel and safe vaccine delivery platform against gut pathogens, such as paratuberculosis, capable of activating both systemic and intestinal immunity.
Asunto(s)
Nanopartículas , Paratuberculosis , Animales , Nanopartículas/química , Paratuberculosis/inmunología , Paratuberculosis/prevención & control , Ratones , Tretinoina/química , Tretinoina/farmacología , Mycobacterium avium subsp. paratuberculosis/inmunología , Copolímero de Ácido Poliláctico-Ácido Poliglicólico/química , Antígenos Bacterianos/inmunología , Antígenos Bacterianos/química , Células Dendríticas/inmunología , Células Dendríticas/efectos de los fármacos , Intestinos/inmunología , Intestinos/microbiología , Ratones Endogámicos C57BL , Femenino , Adyuvantes Inmunológicos/farmacología , Adyuvantes Inmunológicos/química , Adyuvantes Inmunológicos/administración & dosificación , Vacunas Bacterianas/inmunología , Ratones Endogámicos BALB CRESUMEN
Cancer stem cells (CSCs) are one of the determinants of tumor heterogeneity and are characterized by self-renewal, high tumorigenicity, invasiveness, and resistance to various therapies. To overcome the resistance of traditional tumor therapies resulting from CSCs, a strategy of double drug sequential therapy (DDST) for CSC-enriched tumors is proposed in this study and is realized utilizing the developed double-layered hollow mesoporous cuprous oxide nanoparticles (DL-HMCONs). The high drug-loading contents of camptothecin (CPT) and all-trans retinoic acid (ATRA) demonstrate that the DL-HMCON can be used as a generic drug delivery system. ATRA and CPT can be sequentially loaded in and released from CPT3@ATRA3@DL-HMCON@HA. The DDST mechanisms of CPT3@ATRA3@DL-HMCON@HA for CSC-containing tumors are demonstrated as follows: 1) the first release of ATRA from the outer layer induces differentiation from CSCs with high drug resistance to non-CSCs with low drug resistance; 2) the second release of CPT from the inner layer causes apoptosis of non-CSCs; and 3) the third release of Cu+ from DL-HMCON itself triggers the Fenton-like reaction and glutathione depletion, resulting in ferroptosis of non-CSCs. This CPT3@ATRA3@DL-HMCON@HA is verified to possess high DDST efficacy for CSC-enriched tumors with high biosafety.
Asunto(s)
Camptotecina , Cobre , Células Madre Neoplásicas , Humanos , Porosidad , Camptotecina/química , Camptotecina/farmacología , Animales , Cobre/química , Línea Celular Tumoral , Células Madre Neoplásicas/efectos de los fármacos , Tretinoina/química , Tretinoina/farmacología , Nanopartículas/química , Ratones , Antineoplásicos/química , Antineoplásicos/farmacología , Portadores de Fármacos/química , Apoptosis/efectos de los fármacos , Neoplasias/tratamiento farmacológico , Neoplasias/patología , Liberación de FármacosRESUMEN
Recent years have witnessed substantial progress in cancer immunotherapy, specifically T cell-based therapies. However, the application of T cell therapies has been primarily limited to hematologic malignancies, with limited success in the treatment of solid tumors. The main challenge in treating solid tumor is immune escape, which is characterized by reduced antigenicity, diminished immunogenicity, and the development of suppressive tumor immune microenvironments. To address these obstacles and restore T cell-mediated anti-tumor responses, a novel nanoparticle formulation known as PRA@Oxa-c16 is developed. This innovative approach combines retinoic acid and Pt(IV) to specifically target and overcome immune escape. Notably, the therapeutic efficacy of PRA@Oxa-c16 primarily relies on its ability to induce anti-tumor T cell responses, in contrast to the cytotoxicity associated with conventional chemotherapeutic agents. When combined with an immune checkpoint blockade, anti-programmed death-ligand 1 antibody, PRA@Oxa-c16 effectively eliminates solid tumors and induces immune memory responses, which prevent tumor metastasis and recurrence. This promising approach holds great potential for enhancing the treatment of solid tumors with T cell-based immunotherapy.
Asunto(s)
Antígeno B7-H1 , Linfocitos T CD8-positivos , Inmunoterapia , Nanomedicina , Profármacos , Profármacos/química , Profármacos/uso terapéutico , Profármacos/farmacología , Inmunoterapia/métodos , Linfocitos T CD8-positivos/inmunología , Animales , Nanomedicina/métodos , Ratones , Línea Celular Tumoral , Antígeno B7-H1/inmunología , Antígeno B7-H1/antagonistas & inhibidores , Antígeno B7-H1/metabolismo , Nanopartículas/química , Neoplasias/terapia , Neoplasias/inmunología , Humanos , Platino (Metal)/química , Tretinoina/química , Tretinoina/farmacología , Tretinoina/uso terapéutico , Microambiente Tumoral/efectos de los fármacos , Activación de Linfocitos/efectos de los fármacos , Antineoplásicos/química , Antineoplásicos/uso terapéutico , Antineoplásicos/farmacologíaRESUMEN
BACKGROUND AND OBJECTIVE: All-trans retinoic acid (ATRA), an effective differentiation inducer, has been applied clinically to treat acute promyelocytic leukemia (APL). Unfortunately, it is not as potent in other kinds of acute myeloid leukemia (AML). Ethacrynic acid (EA), a classical powerful diuretic, can increase reactive oxygen species (ROS) contents, which can assist ATRA in inducing differentiation in AML cells. Here, we investigated the effect of EA combined with ATRA (EA+RA) on some AML cells except APL. METHODS: Apoptosis and differentiation were determined by morphology, cell viability, Annexin-V assay and CD11c expression. Western blot analysis and the detection of ROS and mitochondrial transmembrane potentials (MMP) were used to investigate the mechanisms. RESULTS: AML cells exhibited differentiation and/or apoptosis after EA+RA treatment. EA+RA increased the intracellular ROS contents. EA+RA-induced apoptosis was accompanied by MMP attenuation and caspase-3/7 activation. EA+RA-induced differentiation was along with MEK/ERK and Akt activation and increased expression of PU.1, CCAAT/enhancer-binding protein ß (C/EBPß) and C/EBPε. N-acetyl-L-cysteine (NAC), an antioxidant, thoroughly reduced EA+RA-increased ROS, and also inhibited MMP attenuation, the activation of caspase- 3/7, MEK/ERK and Akt pathways, the elevation of PU.1 and C/EBPs, and apoptosis and differentiation. However, MEK or PI3K specific inhibitors only suppressed EA+RA-triggered differentiation and the elevation of PU.1 and C/EBPs, but not ROS levels. CONCLUSION: EA+RA induced cell apoptosis through ROS dependent MMP attenuation and caspase 3/7 activation while inducing differentiation by ROS-MEK/ERK-PU.1/C/EBPs and ROS-Akt-PU.1/C/EBPs pathways. In summary, it may provide innovative ATRA-based combination therapy strategies for AML patients via ROS.
Asunto(s)
Antineoplásicos , Apoptosis , Diferenciación Celular , Ensayos de Selección de Medicamentos Antitumorales , Ácido Etacrínico , Leucemia Mieloide Aguda , Especies Reactivas de Oxígeno , Tretinoina , Humanos , Apoptosis/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Tretinoina/farmacología , Tretinoina/química , Diferenciación Celular/efectos de los fármacos , Leucemia Mieloide Aguda/tratamiento farmacológico , Leucemia Mieloide Aguda/patología , Leucemia Mieloide Aguda/metabolismo , Ácido Etacrínico/farmacología , Ácido Etacrínico/química , Antineoplásicos/farmacología , Antineoplásicos/química , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Estructura Molecular , Proliferación Celular/efectos de los fármacos , Relación Estructura-Actividad , Células Tumorales CultivadasRESUMEN
Compound 1 is a potent rexinoid that is highly effective in cancer chemoprevention but elevates serum triglycerides. In an effort to separate the lipid toxicity from the anticancer activity of 1, we synthesized four new analogs of rexinoid 1, of which three rexinoids did not elevate serum triglycerides. Rexinoids 3 and 4 are twice as potent as rexinoid 1 in binding to Retinoid X receptor (RXR). All-trans retinoic acid (ATRA) plays a key role in maintaining skin homeostasis, and rexinoids 3-6 are highly effective in upregulating the genes responsible for the biosynthesis of ATRA. Inflammation plays a key role in skin cancer, and rexinoids 3 and 4 are highly effective in diminishing LPS-induced inflammation. Rexinoids 3 and 4 are highly effective in preventing UVB-induced nonmelanoma skin cancer (NMSC) without displaying any overt toxicities. Biophysical studies of rexinoids 3 and 5 bound to hRXRα-ligand binding domain (LBD) reveal important conformational and dynamical differences in the ligand binding domain.
Asunto(s)
Neoplasias Cutáneas , Tetrahidronaftalenos , Humanos , Tetrahidronaftalenos/química , Ligandos , Receptores X Retinoide/metabolismo , Tretinoina/química , Tretinoina/metabolismo , Neoplasias Cutáneas/tratamiento farmacológico , Neoplasias Cutáneas/prevención & control , Inflamación/tratamiento farmacológico , Inflamación/prevención & control , TriglicéridosRESUMEN
To achieve the co-delivery of doxorubicin (DOX) and small interfering RNA (siRNA) targeting B-cell lymphoma 2 (siBcl-2), lactose acid (LA) and all-trans retinoic acid (ATRA) double grafted N,N,N-trimethyl chitosan (TMC) nanoparticles (GTA NPs) were developed. The relative viability of QGY-7703 cells was decreased to 81.3% when the concentration of GTA NPs was 0.1 mg/mL, but no toxicity to normal cells was observed, indicating that the GTA NPs selectively inhibited the proliferation of tumor cells. With DOX loaded into the hydrophobic core and siRNA condensed onto the hydrophilic shell, GTA/DOX/siRNA NPs were prepared. The GTA/DOX/siRNA NPs possessed high cellular uptake via receptor-mediated endocytosis. Owing to multiple cooperative antitumor effects of DOX, siBcl-2, and GTA NPs, GTA/DOX/siRNA NPs had superior in vitro and in vivo antitumor efficiency to other formulations. These findings provide a guideline for the combined applications of multiple synergistic antitumor manners.
Asunto(s)
Antineoplásicos/farmacología , Quitosano/química , Doxorrubicina/farmacología , Portadores de Fármacos/química , ARN Interferente Pequeño/farmacología , Tretinoina/química , Animales , Antineoplásicos/química , Apoptosis/efectos de los fármacos , Línea Celular Tumoral , Doxorrubicina/química , Sistemas de Liberación de Medicamentos/métodos , Endocitosis/efectos de los fármacos , Femenino , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Ratones , Nanopartículas/química , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , ARN Interferente Pequeño/químicaRESUMEN
In vitro spatiotemporal control of cell differentiation is a critical issue in several biomedical fields such as stem cell therapy and regenerative medicine, as it enables the generation of heterogeneous tissue structures similar to those of their native counterparts. However, the simultaneous control of both spatial and temporal cell differentiation poses important challenges, and therefore no previous studies have achieved this goal. Here, the authors develop a cell differentiation biomolecular electron controller ("Biomoletron") composed of recombinant proteins, DNA, Au nanoparticles, peptides, and an electrically released complex with retinoic acid (RA) to spatiotemporally control SH-SY5Y cell differentiation. RA is only released from the Biomoletron when the complex is electrically stimulated, thus demonstrating the temporal control of SH-SY5Y cell differentiation. Furthermore, by introducing a patterned Au substrate that allows controlling the area where the Biomoletron is immobilized, spatiotemporal differentiation of the SH-SY5Y cell is successfully achieved. Therefore, the proposed Biomoletron-mediated differentiation method provides a promising strategy for spatiotemporal cell differentiation control with applications in regenerative medicine and cell therapy.
Asunto(s)
Azurina/química , ADN/química , Oro/química , Neuronas/citología , Péptidos/química , Tretinoina/farmacología , Diferenciación Celular/efectos de los fármacos , Línea Celular , Tratamiento Basado en Trasplante de Células y Tejidos , Péptidos de Penetración Celular/química , Fenómenos Electromagnéticos , Humanos , Nanopartículas del Metal , Neuronas/efectos de los fármacos , Oligopéptidos/química , Medicina Regenerativa , Análisis Espacio-Temporal , Tretinoina/químicaRESUMEN
Here, we found that all-trans retinoic acid (ATRA), the most biologically active metabolite of vitamin A, strengthens the anti-viral defense mechanism of E6-associated protein (E6AP) that downregulates hepatitis C virus (HCV) Core levels via ubiquitin-dependent proteasomal degradation. For this effect, ATRA downregulated both protein and enzyme activity levels of DNA methyltransferase 1 and 3b and activated E6AP expression via promoter hypomethylation in HepG2 cells but not in Hep3B cells, in which p53 was absent. Ectopic p53 expression but not E6AP overexpression restored the ability of ATRA to downregulate HCV Core levels in Hep3B cells, suggesting a direct role of p53 in the E6AP-mediated ubiquitination of HCV Core. ATRA also downregulated HCV Core levels during HCV infection in Huh7D cells to inhibit virus replication, providing theoretical basis for the clinical application of ATRA against HCV infection.
Asunto(s)
Regulación hacia Abajo , Hepacivirus/metabolismo , Hepatitis C/metabolismo , Tretinoina/química , Metilación de ADN , Relación Dosis-Respuesta a Droga , Células Hep G2 , Humanos , Complejo de la Endopetidasa Proteasomal/metabolismo , Proteolisis , Ubiquitina/metabolismo , Ubiquitina-Proteína Ligasas/metabolismo , Ubiquitinación/efectos de los fármacos , Proteínas del Núcleo Viral/metabolismo , Replicación Viral/efectos de los fármacosRESUMEN
Here, we assessed the efficacy and safety of Nano lipid carrier (NLC) drug delivery system containing tretinoin (NLC-TRE) in comparison with the conventional 0.05% tretinoin cream (TRE cream) in mild to moderate acne vulgaris. A stable and appropriate NLC-TRE formulation was prepared using a high-pressure homogenizer and particle characterization and physicochemical properties were evaluated under accelerated conditions. Efficacy assessment was performed via a split-face clinical study, by comparing the number of acne lesions, porphyrin production and skin biophysical parameters in both sides of the face randomly treated with NLC-TRE and TRE cream. Plasma concentration of tretinoin after topical application of NLC-TRE was measured for primary safety evaluation. We acquired a stable, spherical nanoparticles with particle size of 118.5 nm, PI equal to 0.485 and ZP of - 44.7 mV. The rate of decrease of acne lesions was significantly higher in NLC- TRE side (p value < 0.001). The size and intensity of porphyrin production in pilosebaceous follicles were significantly reduced only on NLC-TRE side (p value < 0.01). The plasma concentration of the tretinoin, after 8 weeks' application remained lower than the toxic levels. The NLC-TRE formula provides better efficiency and good loading capacity of TRE in the drug delivery system.
Asunto(s)
Acné Vulgar , Porfirinas , Acné Vulgar/tratamiento farmacológico , Acné Vulgar/patología , Método Doble Ciego , Portadores de Fármacos , Humanos , Piel/patología , Tretinoina/química , Tretinoina/uso terapéuticoRESUMEN
Cationic nanocapsules represent a promising approach for topical delivery purposes. We elaborated on a novel formulation based on the cationic nanocapsules to enhance the pharmacodynamic efficacy, user compliance, and photostability of tretinoin (TTN). To achieve this goal, TTN nanocapsules were prepared by the nanoprecipitation method. In order to statistically optimize formulation variables, a Box-Behnken design, using Design-Expert software, was employed. Three independent variables were evaluated: total weight of the cationic acrylic polymer (X 1), oil volume (X 2), and TTN amount (X 3). The particle size and encapsulation efficiency percent (EE%) were selected as dependent variables. The optimal formulation demonstrated spherical morphology under scanning electron microscopy (SEM), optimum particle size of 116.3 nm, and high EE% of 83.2%. TTN-loaded nanocapsules improved photostability compared to its methanolic solution. The in vitro release study data showed that tretinoin was released in a sustained manner compared to the free drug. The ex vivo skin permeation study demonstrated that greater drug deposition into the epidermal region rather than the deep skin was observed with a gel containing TTN-loaded nanocapsules than that of drug solution, respectively. The skin irritation test revealed that the nanoencapsulation of the drug decreased its irritancy compared to the free drug. These results revealed the promising potential of cationic nanocapsules for topical delivery of tretinoin.
Asunto(s)
Cationes/química , Nanocápsulas/química , Piel/metabolismo , Tretinoina/administración & dosificación , Tretinoina/química , Administración Cutánea , Animales , Química Farmacéutica/métodos , Portadores de Fármacos/química , Composición de Medicamentos/métodos , Sistemas de Liberación de Medicamentos/métodos , Masculino , Tamaño de la Partícula , Ratas , Ratas Wistar , Absorción Cutánea/fisiología , Solubilidad/efectos de los fármacosRESUMEN
Despite recent advances in the treatment of human colon cancer, the chemotherapeutic efficacy against colon cancer is still unsatisfactory. The complexity in colorectal cancer treatment leads to new research in combination therapy to overcome multidrug resistance in cancer and increase apoptosis. The objective of the present research work was to develop polyplexes for co-delivery of plasmid DNA with retinoic acid against colorectal cancer cell line (HCT-15). Plain polyplexes were prepared using chitosan and hyaluronic acid solution (0.1% w/v), whereas retinoic acid polyplexes were prepared using ethanol: water (1:9 v/v) system. The particle size was observed in the order of chitosan solution > blank polyplex > retinoic acid-loaded polyplex. Encapsulation efficiency of retinoic acid was found to be 81.51 ± 4.33% for retinoic acid-loaded polyplex formulation. The drug release was observed to be in a controlled pattern with 72.23 ± 1.32% release of retenoic acid from polyplex formulation. Cell line studies of the formulation displayed better cell inhibition and low cytotoxicity for the retinoic acid-loaded polyplexes in comparison to pure retinoic acid, thus demonstrating better potential action against colorectal cancer cell line HCT-15. Retinoic acid-loaded polyplexes indicated higher potential for the delivery of the active whereas the cell line studies displayed the efficacy of the formulation against colorectal cancer cell line HCT-15.
Asunto(s)
Neoplasias Colorrectales/tratamiento farmacológico , Portadores de Fármacos , Nanoestructuras/química , Tretinoina/administración & dosificación , Línea Celular Tumoral , Neoplasias Colorrectales/patología , Portadores de Fármacos/síntesis química , Portadores de Fármacos/química , Portadores de Fármacos/farmacocinética , Composición de Medicamentos/métodos , Liberación de Fármacos , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Ácido Hialurónico/síntesis química , Ácido Hialurónico/química , Ácido Hialurónico/farmacocinética , Nanoestructuras/uso terapéutico , Tamaño de la Partícula , Polímeros/química , Polímeros/farmacología , Espectroscopía Infrarroja por Transformada de Fourier , Tretinoina/química , Tretinoina/farmacocinéticaRESUMEN
The vitamin A derivative, retinoid acid (RA) is key player in guiding adaptive mucosal immune responses. However, data on the uptake and metabolism of vitamin A within human immune cells has remained largely elusive because retinoids are small, lipophilic molecules which are difficult to detect. To overcome this problem and to be able to study the effect of vitamin A metabolism in human immune cell subsets, we have synthesized novel bio-orthogonal retinoid-based probes (clickable probes), which are structurally and functionally indistinguishable from vitamin A. The probes contain a functional group (an alkyne) to conjugate to a fluorogenic dye to monitor retinoid molecules in real-time in immune cells. We demonstrate, by using flow cytometry and microscopy, that multiple immune cells have the capacity to internalize retinoids to varying degrees, including human monocyte-derived dendritic cells (DCs) and naïve B lymphocytes. We observed that naïve B cells lack the enzymatic machinery to produce RA, but use exogenous retinoic acid to enhance CD38 expression. Furthermore, we showed that human DCs metabolize retinal into retinoic acid, which in co-culture with naïve B cells led to of the induction of CD38 expression. These data demonstrate that in humans, DCs can serve as an exogenous source of RA for naïve B cells. Taken together, through the use of clickable vitamins our data provide valuable insight in the mechanism of vitamin A metabolism and its importance for human adaptive immunity.
Asunto(s)
Linfocitos B/inmunología , Química Clic/métodos , Células Dendríticas/inmunología , Vitamina A/metabolismo , ADP-Ribosil Ciclasa 1/metabolismo , Inmunidad Adaptativa , Células Cultivadas , Técnicas de Cocultivo , Cobre/metabolismo , Citometría de Flujo , Colorantes Fluorescentes , Humanos , Tretinoina/química , Tretinoina/metabolismo , Regulación hacia Arriba , Vitamina A/químicaRESUMEN
Glioblastoma multiforme (GBM) is the most aggressive glioma. The treatment response is always low, and the condition is typically rapidly fatal. The undifferentiated and self-renewal characteristics of cancer stem cells (CSCs) have been reported, and their potential contribution may cause tumor initiation, recurrence, metastasis, and therapeutic resistance. In particular, glioblastoma stem-like cells exhibit highly invasive properties and drug resistance, serving as a model for the development of novel therapeutic strategies. Induction therapy provides an alternative therapeutic strategy to eliminate the stem cell properties of CSCs and enhance therapeutic sensitivity. The differentiated cells may lose their self-renewal ability, downregulate stem cell-related genes and drug resistance genes, and enhance anticancer drug sensitivity. Therefore, the purpose of this study is to establish a niche for glioblastoma stem-like cell selection as a platform and facilitate the assessment of differentiation therapy on GBM cancer stem-like colonies by retinoic acid (RA) with temozolomide (TMZ)-loaded gold nanoparticles (GNPs) associated with low-intensity ultrasound (LIUS). Herein, a hyaluronic acid-based material system was used to isolate GBM cancer stem-like colonies. Colony formation, size determination, stem cell-related marker expression, and GBM cancer stem-like cell marker expression with the culture period were identified. The effect of TMZ on GBM stem-like colonies on HA-based material systems was also determined, and the results revealed that drug resistance was highly enhanced in GBM colonies compared with that in the control cell population. In addition, GBM colonies also exhibited a significant increase in breast cancer resistance protein expression, which is consistent with the drug resistance effect. Furthermore, several factors, including LIUS, RA, and GNPs, were used to determine the possibility of induction therapy. RA with TMZ-loaded GNP-associated LIUS stimulation exhibited a significant and synergistic effect on the differentiation effect and drug sensitivity enhancement. The GBM cancer stem-like colony system presents an opportunity for the development of new therapeutic strategies, and this study provides an alternative differentiation therapy for malignant tumors.
Asunto(s)
Antineoplásicos/farmacología , Portadores de Fármacos/química , Glioblastoma/tratamiento farmacológico , Nanopartículas del Metal/química , Temozolomida/farmacología , Tretinoina/farmacología , Antineoplásicos/química , Diferenciación Celular/efectos de los fármacos , Línea Celular Tumoral , Liberación de Fármacos , Resistencia a Antineoplásicos/efectos de los fármacos , Sinergismo Farmacológico , Oro/química , Humanos , Ácido Hialurónico/química , Quimioterapia de Inducción , Células Madre Neoplásicas/efectos de los fármacos , Esferoides Celulares/efectos de los fármacos , Temozolomida/química , Tretinoina/química , Ondas UltrasónicasRESUMEN
Differentiation therapy is attracting increasing interest in cancer as it can be more specific than conventional chemotherapy approaches, and it has offered new treatment options for some cancer types, such as treating acute promyelocytic leukaemia (APL) by retinoic acid. However, there is a pressing need to identify additional molecules which act in this way, both in leukaemia and other cancer types. In this work, we hence developed a novel transcriptional drug repositioning approach, based on both bioinformatics and cheminformatics components, that enables selecting such compounds in a more informed manner. We have validated the approach for leukaemia cells, and retrospectively retinoic acid was successfully identified using our method. Prospectively, the anti-parasitic compound fenbendazole was tested in leukaemia cells, and we were able to show that it can induce the differentiation of leukaemia cells to granulocytes in low concentrations of 0.1 µM and within as short a time period as 3 days. This work hence provides a systematic and validated approach for identifying small molecules for differentiation therapy in cancer.