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1.
Eur J Med Chem ; 216: 113315, 2021 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-33711763

RESUMO

The synthesis and in vitro anti-HIV activity of a novel series of phosphoramidate pronucleotides including a S-pivaloyl-2-thioethyl (tBuSATE) group as biolabile phosphate protecting group are reported. Such constructs, obtained through different phosphorus chemistries, are characterized by the association of two different anti-HIV nucleoside analogues linked to the phosphorus atom respectively by the sugar residue and the exocyclic amino function of the nucleobase. In vitro, comparative anti-HIV evaluation demonstrates that such original prodrugs are able to allow the efficient intracellular combination release of a 5'-mononucleotide as well as another nucleoside analogue. In human T4-lymphoblastoid cells, the pronucleotide 1 shows remarkable antiviral activity with an EC50 in the nanomolar range (0.6 ηM) and without additional cytotoxicity. In addition, these two pronucleotide models exhibit higher selectivity index than the equimolar mixture of their constitutive nucleoside analogues opening the way to further studies with regard to the current use of drug combinations.


Assuntos
Nucleotídeos/química , Fármacos Anti-HIV/química , Fármacos Anti-HIV/metabolismo , Fármacos Anti-HIV/farmacologia , Antivirais/química , Antivirais/metabolismo , Antivirais/farmacologia , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Portadores de Fármacos/química , HIV-1/efeitos dos fármacos , Humanos , Nucleotídeos/metabolismo , Nucleotídeos/farmacologia , Pró-Fármacos/química , Pró-Fármacos/metabolismo , Pró-Fármacos/farmacologia
2.
Eur J Med Chem ; 215: 113276, 2021 Apr 05.
Artigo em Inglês | MEDLINE | ID: mdl-33611186

RESUMO

A series of novel amphiphilic paclitaxel (PTX) small molecule prodrugs, PTX-succinic anhydride-cystamine (PTX-Cys), PTX-dithiodipropionic anhydride (PTX-SS-COOH) and PTX-succinic anhydride-cystamine-valine (PTX-SS-Val) were designed, synthesized and evaluated against cancer cell lines. Compared with paclitaxel, these prodrugs contained water-soluble groups such as amino, carboxyl and amino acid, which improved the aqueous solubility of the prodrugs. More importantly, the valine was introduced in PTX-SS-Val molecule and made the molecule conform to the structural characteristics of intestinal oligopeptide transporter PEPT1 substrate. Thus the oral bioavailability of prodrug could be improved because of the mediation of PEPT1 transporter. These small molecule paclitaxel prodrugs could self-assemble into nanoparticles in aqueous solution, which effectively improved the solubility of paclitaxel, and had certain stability in pH 6.5, pH 7.4 buffer solutions and simulated gastrointestinal fluids. Some of these prodrugs, especially for PTX-Cys and PTX-SS-Val, exhibited nearly equal or slightly better anticancer activity when compared to paclitaxel. Further studies on PTX-Cys and PTX-SS-Val showed that both had good intestinal absorption in the rat single-pass intestinal perfusion (SPIP) experiments. Oral pharmacokinetic experiments showed that PTX-SS-Val could effectively improve the oral bioavailability of PTX.


Assuntos
Antineoplásicos Fitogênicos/farmacocinética , Absorção Intestinal/fisiologia , Paclitaxel/farmacocinética , Pró-Fármacos/farmacocinética , Tensoativos/farmacocinética , Animais , Antineoplásicos Fitogênicos/síntese química , Antineoplásicos Fitogênicos/metabolismo , Estabilidade de Medicamentos , Humanos , Mucosa Intestinal/metabolismo , Células MCF-7 , Masculino , Nanopartículas/química , Nanopartículas/metabolismo , Paclitaxel/síntese química , Paclitaxel/metabolismo , Transportador 1 de Peptídeos/metabolismo , Pró-Fármacos/síntese química , Pró-Fármacos/metabolismo , Ratos Wistar , Tensoativos/síntese química , Tensoativos/metabolismo
3.
Eur J Med Chem ; 215: 113288, 2021 Apr 05.
Artigo em Inglês | MEDLINE | ID: mdl-33640763

RESUMO

Kinesin spindle protein (KSP) is expressed only in cells undergoing cell division, and hence represents an attractive target for the treatment of cancer. Several KSP inhibitors have been developed and undergone clinical trial, but their clinical use is limited by their toxicity to rapidly proliferating non-cancerous cells. To create new KSP inhibitors that are highly selective for cancer cells, we optimized the amino acid moiety of S-trityl-l-cysteine (STLC) derivative 1 using in silico modeling. Molecular docking and molecular dynamics simulation were performed to investigate the binding mode of 1 with KSP. Consistent with the structure activity relationship studies, we found that a cysteine amino moiety plays an important role in stabilizing the interaction. Based on these findings and the structure of GSH, a substrate of γ-glutamyltransferase (GGT), we designed and synthesized the prodrug N-γ-glutamylated STLC derivative 9, which could be hydrolyzed by GGT to produce 1. The KSP ATPase inhibitory activity of 9 was lower than that of 1, and LC-MS analysis indicated that 9 was converted to 1 only in the presence of GGT in vitro. In addition, the cytotoxic activity of 9 was significantly attenuated in GGT-knockdown A549 cells. Since GGT is overexpressed on the cell membrane of various cancer cells, these results suggest that compound 9 could be a promising prodrug that selectively inhibits the proliferation of GGT-expressing cancer cells.


Assuntos
Antineoplásicos/farmacologia , Cisteína/farmacologia , Dibenzocicloeptenos/farmacologia , Cinesina/antagonistas & inibidores , Pró-Fármacos/farmacologia , Compostos de Tritil/farmacologia , Animais , Antineoplásicos/síntese química , Antineoplásicos/metabolismo , Bovinos , Linhagem Celular Tumoral , Cisteína/síntese química , Cisteína/metabolismo , Dibenzocicloeptenos/síntese química , Dibenzocicloeptenos/metabolismo , Humanos , Cinesina/metabolismo , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Estrutura Molecular , Pró-Fármacos/síntese química , Pró-Fármacos/metabolismo , Ligação Proteica , Relação Estrutura-Atividade , Termodinâmica , Compostos de Tritil/síntese química , Compostos de Tritil/metabolismo , gama-Glutamiltransferase/metabolismo
4.
Carbohydr Polym ; 255: 117393, 2021 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-33436222

RESUMO

In this paper, a novel redox-responsive nanoparticles has been designed for targeted delivery of docetaxel (DTX). Chondroitin sulfate (CS) was used to construct the nanoparticles due to the ability of tumor targeting through binding with CD44 receptor that overexpresses on the surfaces of various tumor cells. A redox-responsive small-molecular DTX prodrug was prepared through modifying with cystamine containing disulfide bonds (Cys-DTX). Then the DTX prodrug was grafted to the CS to construct the amphiphilic polymer (CS-ss-DTX). Further, Cys-DTX/CS-ss-DTX nanoparticles were formed by self-assembly of amphiphilic polymer and incorporation of free Cys-DTX prodrug. This category of nanosized DTX delivery system was expected for not only exhibiting high permeability and cytotoxicity of Cys-DTX prodrug, but also targeting transportation of encapsulated redox-responsive Cys-DTX prodrug. According to results of related researches on physicochemical properties and biological evaluation, the novel redox-responsive Cys-DTX/CS-ss-DTX nanoparticles increased amount of DTX released from the nanoparticles in reductive environment, improved permeability in tumor tissues, enhanced cytotoxicity and decreased side effects compared with free DTX. All of these results showed that this kind of Cys-DTX/CS-ss-DTX nanoparticles were worthy of being expectation in tumor chemotherapy in future.


Assuntos
Antineoplásicos/farmacologia , Sulfatos de Condroitina/química , Docetaxel/farmacologia , Glicoconjugados/farmacologia , Melanoma Experimental/tratamento farmacológico , Pró-Fármacos/farmacologia , Neoplasias Cutâneas/tratamento farmacológico , Animais , Antineoplásicos/química , Antineoplásicos/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Docetaxel/química , Docetaxel/metabolismo , Portadores de Fármacos/síntese química , Portadores de Fármacos/metabolismo , Portadores de Fármacos/farmacologia , Liberação Controlada de Fármacos , Feminino , Glicoconjugados/química , Glicoconjugados/metabolismo , Humanos , Células MCF-7 , Melanoma Experimental/metabolismo , Melanoma Experimental/patologia , Camundongos , Micelas , Nanopartículas/administração & dosagem , Nanopartículas/química , Nanopartículas/ultraestrutura , Oxirredução , Pró-Fármacos/química , Pró-Fármacos/metabolismo , Neoplasias Cutâneas/metabolismo , Neoplasias Cutâneas/patologia , Carga Tumoral/efeitos dos fármacos
5.
ACS Appl Mater Interfaces ; 13(1): 245-256, 2021 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-33373182

RESUMO

Carrier-free pure drug self-assembled nanosystems have been proposed as a promising strategy for synergetic anticancer therapy. Herein, we purposefully designed and synthesized disulfide-modified glutathione (GSH)-responsive natural pentacyclic triterpene betulinic acid (BA) with better biodegradability and biocompatibility to construct carrier-free photosensitive prodrugs BA-S-S/Ce6 NPs for synergistically enhanced and biosafe photochemotherapy. The molecular dynamics simulation elucidates the possible coassembly mechanism that the coplanar arrangement of BA-S-S dimeric may be primarily responsible for the formation of a long lamella-like or spherical morphology. The density functional theory calculations demonstrate that the reduced energy gap (ΔEST) of Ce6 facilitates the improved singlet oxygen generation of BA-S-S/Ce6 nanoparticles (NPs). The assembled prodrugs exhibited remarkable GSH-responsive property and multiple favorable therapeutic features, leading to enhanced synergistic antitumor efficacy without noticeable toxicity. Additionally, evaluation of the antitumor efficacy of another tetracyclic triterpene stigmasterol (ST)-mediated ST-S-S/Ce6 NPs further confirmed the effectiveness of this rational design. This work provides a promising insight for exploring the pure drug self-assembly behavior and construction of GSH-responsive carrier-free triterpenoid prodrugs toward improved multiple combination antitumor therapies.


Assuntos
Antineoplásicos/uso terapêutico , Glutationa/metabolismo , Nanopartículas/uso terapêutico , Neoplasias/tratamento farmacológico , Triterpenos Pentacíclicos/uso terapêutico , Pró-Fármacos/uso terapêutico , Animais , Antineoplásicos/química , Antineoplásicos/metabolismo , Teoria da Densidade Funcional , Sinergismo Farmacológico , Feminino , Luz , Camundongos Endogâmicos BALB C , Modelos Químicos , Simulação de Dinâmica Molecular , Nanopartículas/química , Triterpenos Pentacíclicos/química , Triterpenos Pentacíclicos/metabolismo , Fotoquimioterapia , Fármacos Fotossensibilizantes/química , Fármacos Fotossensibilizantes/uso terapêutico , Porfirinas/química , Porfirinas/uso terapêutico , Pró-Fármacos/química , Pró-Fármacos/metabolismo , Oxigênio Singlete/metabolismo , Estigmasterol/análogos & derivados , Estigmasterol/metabolismo , Estigmasterol/uso terapêutico
6.
J Med Chem ; 63(24): 15785-15801, 2020 12 24.
Artigo em Inglês | MEDLINE | ID: mdl-33320012

RESUMO

Mutations in the human PANK2 gene are implicated in neurodegenerative diseases such as pantothenate kinase-associated neurodegeneration (PKAN) and result in low levels of coenzyme-A (CoA) in the CNS due to impaired production of phosphopantothenic acid (PPA) from vitamin B5. Restoration of central PPA levels by delivery of exogenous PPA is a recent strategy to reactivate CoA biosynthesis in PKAN patients. Fosmetpantotenate is an oral PPA prodrug. We report here the development of a new PANk2-/- knockout model that allows CoA regeneration in brain cells to be evaluated and describe two new series of cyclic phosphate prodrugs of PPA capable of regenerating excellent levels of CoA in this system. A proof-of-concept study in mouse demonstrates the potential of this new class of prodrugs to deliver PPA to the brain following oral administration and confirms incorporation of the prodrug-derived PPA into CoA.


Assuntos
Ácido Pantotênico/análogos & derivados , Pró-Fármacos/química , Animais , Barreira Hematoencefálica/efeitos dos fármacos , Barreira Hematoencefálica/metabolismo , Encéfalo/metabolismo , Coenzima A/metabolismo , Ciclização , Modelos Animais de Doenças , Meia-Vida , Hepatócitos/citologia , Hepatócitos/metabolismo , Humanos , Gotículas Lipídicas/química , Gotículas Lipídicas/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Neurodegeneração Associada a Pantotenato-Quinase/tratamento farmacológico , Neurodegeneração Associada a Pantotenato-Quinase/patologia , Ácido Pantotênico/química , Ácido Pantotênico/metabolismo , Ácido Pantotênico/uso terapêutico , Pró-Fármacos/metabolismo , Pró-Fármacos/uso terapêutico , Relação Estrutura-Atividade
7.
J Med Chem ; 63(13): 6991-7007, 2020 07 09.
Artigo em Inglês | MEDLINE | ID: mdl-32515595

RESUMO

The antiviral efficacy of many nucleoside analogues is strongly dependent on their intracellular activation by host cellular kinases to yield ultimately the bioactive nucleoside analogue triphosphates (NTP). The metabolic conversion of nucleoside analogues into their triphosphates often proceeds insufficiently. We developed a nucleoside triphosphate (NTP) delivery system (the TriPPPro approach), in which the γ-phosphate is covalently modified by two different biodegradable masking units, one is the acyloxybenzyl (AB) moiety and the other is the alkoxycarbonyloxybenzyl (ACB) group. Such compounds formed NTPs with high selectivity by an enzyme-triggered mechanism in human T-lymphocyte CEM cell extracts loosing first the AB moiety, followed by the ACB group. This enables the bypass of all steps of the intracellular phosphorylation. This approach was applied here to convert some modestly active or even inactive nucleoside analogues into powerful biologically active metabolites. Potent antiviral activity profiles were obtained depending on the lipophilicity of the TriPPPro-NTP prodrugs against HIV-1 and HIV-2 replication in cultures of infected wild-type CD4+ CEM T-cells and more importantly in thymidine kinase-deficient CD4+ T-cells (CEM/TK-). This TriPPPro strategy offers high potential for future antiviral and antitumoral chemotherapies.


Assuntos
Interações Hidrofóbicas e Hidrofílicas , Nucleosídeos/química , Nucleosídeos/metabolismo , Polifosfatos/química , Pró-Fármacos/química , Pró-Fármacos/metabolismo , Fármacos Anti-HIV/química , Fármacos Anti-HIV/metabolismo , Fármacos Anti-HIV/farmacologia , HIV-1/efeitos dos fármacos , HIV-1/fisiologia , HIV-2/efeitos dos fármacos , HIV-2/fisiologia , Humanos , Nucleosídeos/farmacologia , Replicação Viral/efeitos dos fármacos
8.
J Med Chem ; 63(11): 6003-6027, 2020 06 11.
Artigo em Inglês | MEDLINE | ID: mdl-32421343

RESUMO

We disclose a study on nucleoside triphosphate (NTP) analogues in which the γ-phosphate is covalently modified by two different biodegradable masking units and d4T as nucleoside analogue that enable the delivery of d4TTP with high selectivity in phosphate buffer (pH 7.3) and by enzyme-triggered reactions in human CD4+ T-lymphocyte CEM cell extracts. This allows the bypass of all steps normally needed in the intracellular phosphorylation. These TriPPPro-nucleotides comprising an acyloxybenzyl (AB; ester) or an alkoxycarbonyloxybenzyl (ACB; carbonate) in combination with an ACB moiety are described as NTP delivery systems. The introduction of these two different groups led to the selective formation of γ-(ACB)-d4TTPs by chemical hydrolysis and in particular by cell extract enzymes. γ-(AB)-d4TTPs are faster cleaved than γ-(ACB)-d4TTPs. In antiviral assays, the compounds are highly active against HIV-1 and HIV-2 in wild-type CEM/O cells and more importantly in thymidine kinase-deficient CD4+ T-cells (CEM/TK-).


Assuntos
Fármacos Anti-HIV/farmacologia , HIV-1/efeitos dos fármacos , HIV-2/efeitos dos fármacos , Nucleotídeos/química , Pró-Fármacos/química , Animais , Fármacos Anti-HIV/química , Fármacos Anti-HIV/farmacocinética , Linfócitos T CD4-Positivos/citologia , Linfócitos T CD4-Positivos/metabolismo , Esterases/metabolismo , Meia-Vida , Humanos , Hidrólise , Fígado/enzimologia , Nucleotídeos/metabolismo , Nucleotídeos/farmacologia , Pró-Fármacos/metabolismo , Pró-Fármacos/farmacologia , Suínos
9.
J Med Chem ; 63(13): 6741-6747, 2020 07 09.
Artigo em Inglês | MEDLINE | ID: mdl-32410451

RESUMO

Effective delivery to the brain limits the development of novel glioblastoma therapies. Here, we introduce conjugation between platinum(IV) prodrugs of cisplatin and perfluoroaryl peptide macrocycles to increase brain uptake. We demonstrate that one such conjugate shows efficacy against glioma stem-like cells. We investigate the pharmacokinetics of this conjugate in mice and show that the amount of platinum in the brain after treatment with the conjugate is 15-fold greater than with cisplatin after 5 h.


Assuntos
Encéfalo/metabolismo , Compostos Macrocíclicos/química , Peptídeos/química , Platina/química , Platina/metabolismo , Pró-Fármacos/metabolismo , Transporte Biológico , Linhagem Celular , Humanos
10.
Am J Physiol Cell Physiol ; 319(1): C129-C135, 2020 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-32374677

RESUMO

The purpose of this study was to evaluate a new pharmacological strategy using a first-generation succinate prodrug, NV118, in peripheral blood mononuclear cells (PBMCs) obtained from subjects with carbon monoxide (CO) poisoning and healthy controls. We obtained human blood cells from subjects with CO poisoning and healthy control subjects. Intact PBMCs from subjects in the CO and Control group were analyzed with high-resolution respirometry measured in pmol O2 per second per 10-6 PBMCs. In addition to obtaining baseline respiration, NV118 (100 µM) was injected, and the same parameters of respiration were obtained for comparison in PBMCs. We measured mitochondrial dynamics with microscopy with the same conditions. We enrolled 37 patients (17 in the CO group and 20 in the Control group for comparison) in the study. PMBCs obtained from subjects in the CO group had overall significantly lower respiration compared with the Control group (P < 0.0001). There was a significant increase in respiration with NV118, specifically with an increase in maximum respiration and respiration from complex II and complex IV (P < 0.0001). The mitochondria in PBMCs demonstrated an overall increase in net movement compared with the Control group. Our results of this study suggest that the therapeutic compound, NV118, increases respiration at complex II and IV as well as restoration of mitochondrial movement in PBMCs obtained from subjects with CO poisoning. Mitochondrial-directed therapy offers a potential future strategy with further exploration in vivo.


Assuntos
Intoxicação por Monóxido de Carbono/metabolismo , Permeabilidade da Membrana Celular/fisiologia , Leucócitos Mononucleares/metabolismo , Mitocôndrias/metabolismo , Pró-Fármacos/metabolismo , Ácido Succínico/metabolismo , Permeabilidade da Membrana Celular/efeitos dos fármacos , Respiração Celular/efeitos dos fármacos , Respiração Celular/fisiologia , Humanos , Leucócitos Mononucleares/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Pró-Fármacos/administração & dosagem , Ácido Succínico/administração & dosagem
11.
J Med Chem ; 63(9): 4732-4748, 2020 05 14.
Artigo em Inglês | MEDLINE | ID: mdl-32275415

RESUMO

Screening of a GSK-proprietary library against intracellular Mycobacterium tuberculosis identified 1, a thioalkylbenzoxazole hit. Biological profiling and mutant analysis revealed that this compound is a prodrug that is bioactivated by the mycobacterial enzyme MymA. A hit-expansion program including design, synthesis, and profiling of a defined set of analogues with optimized drug-like properties led to the identification of an emerging lead compound, displaying potency against intracellular bacteria in the low micromolar range, high in vitro solubility and permeability, and excellent microsomal stability.


Assuntos
Antituberculosos/farmacologia , Proteínas de Bactérias/metabolismo , Benzoxazóis/farmacologia , Mycobacterium tuberculosis/efeitos dos fármacos , Oxigenases/metabolismo , Pró-Fármacos/farmacologia , Animais , Antituberculosos/síntese química , Antituberculosos/metabolismo , Benzoxazóis/síntese química , Benzoxazóis/metabolismo , Linhagem Celular Tumoral , Humanos , Camundongos , Testes de Sensibilidade Microbiana , Microssomos Hepáticos/efeitos dos fármacos , Estrutura Molecular , Pró-Fármacos/síntese química , Pró-Fármacos/metabolismo , Relação Estrutura-Atividade
12.
Org Biomol Chem ; 18(15): 2877-2885, 2020 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-32236231

RESUMO

The nutraceutical Nicotinamide Riboside (NR), an efficacious biosynthetic precursor to NAD, is readily metabolized by the purine nucleoside phosphorylase (PNP). Access to the PNP-stable versions of NR is difficult because the glycosidic bond of NR is easily cleaved. Unlike NR, NRH, the reduced form of NR, offers sufficient chemical stability to allow the successful functionalisation of the ribosyl-moiety. Here, we report on a series of NRH and NR derived amino acid conjugates, generated in good to excellent yields and show that O5'-esterification prevents the PNP-catalyzed phosphorolysis of these NR prodrugs.


Assuntos
Aminoácidos/metabolismo , Niacinamida/análogos & derivados , Pró-Fármacos/metabolismo , Purina-Núcleosídeo Fosforilase/metabolismo , Aminoácidos/química , Biocatálise , Estrutura Molecular , Niacinamida/química , Niacinamida/metabolismo , Pró-Fármacos/química , Purina-Núcleosídeo Fosforilase/química
13.
Yakugaku Zasshi ; 140(3): 369-376, 2020.
Artigo em Japonês | MEDLINE | ID: mdl-32115554

RESUMO

The first-pass hydrolysis of oral ester-type prodrugs in the liver and intestine is mediated mainly by hCE1 and hCE2 of the respective predominant carboxylesterase (CES) isozymes. In order to provide high blood concentrations of the parent drugs, it is preferable that prodrugs are absorbed as an intact ester in the intestine, then rapidly converted to active parent drugs by hCE1 in the liver. In the present study, we designed a prodrug of fexofenadine (FXD) as a model parent drug that is resistant to hCE2 but hydrolyzed by hCE1, utilizing the differences in catalytic characteristics of hCE1 and hCE2. In order to precisely predict the intestinal absorption of an FXD prodrug candidate, we developed a novel high-throughput system by modifying Caco-2 cells. Further, we evaluated species differences and aging effects in the intestinal and hepatic hydrolysis of prodrugs to improve the estimation of in vivo first-pass hydrolysis of ester-type prodrugs. Consequently, it was possible to design a hepatotropic prodrug utilizing the differences in tissue distribution and substrate specificity of CESs. In addition, we successfully established three useful in vitro systems for predicting the intestinal absorption of hCE1 substrate using Caco-2 cells. However, some factors involved in estimating the bioavailability of prodrugs in human, such as changes in recognition of drug transporters by esterification, and species differences of the first-pass hydrolysis, should be comprehensively considered in prodrug development.


Assuntos
Ésteres/metabolismo , Pró-Fármacos/metabolismo , Administração Oral , Disponibilidade Biológica , Hidrolases de Éster Carboxílico/fisiologia , Ésteres/administração & dosagem , Humanos , Hidrólise , Absorção Intestinal , Isoenzimas/fisiologia , Fígado/metabolismo , Pró-Fármacos/administração & dosagem , Especificidade da Espécie
14.
ACS Appl Mater Interfaces ; 12(10): 11307-11319, 2020 Mar 11.
Artigo em Inglês | MEDLINE | ID: mdl-32048820

RESUMO

Gene-directed enzyme-prodrug therapy (GDEPT) is a promising approach for cancer therapy, but it suffers from poor targeted delivery in vivo. Polyethylenimine (PEI) is a cationic polymer efficient in delivering negatively charged nucleic acids across cell membranes; however, it is highly toxic in vivo. Hence, we efficiently reduced PEI toxicity without compromising its transfection efficiency by conjugating it with poly(d,l-lactic-co-glycolic acid) (PLGA) and poly(ethylene glycol) (PEG) as triblock copolymers through a multistep synthetic process. The synthesized nanoparticles showed efficient delivery of loaded nucleic acids to tumor cells in vitro and in vivo in mice. We used this nanoparticle to deliver a rationally engineered thymidine kinase (TK)-p53-nitroreductase (NTR) triple therapeutic gene against hepatocellular carcinoma (HCC), where p53 tumor suppressor gene is mutated in more than 85% of cancers. TK-p53-NTR triple gene therapy restores p53 function and potentiates cancer cell response to delivered prodrugs (ganciclovir (GCV) and CB1954). We used SP94 peptide-functionalized PLGA-PEG-PEI nanoparticles for the optimal delivery of TK-p53-NTR therapeutic gene in vivo. The nanoparticles prepared from the conjugated polymer showed high loading efficiency for the DNA and markedly enhanced TK-NTR-mediated gene therapy upon the simultaneous coexpression of p53 by the concurrent rescue of the endogenous apoptotic pathway in HCC cells of both p53-mutant and wild-type phenotypes in vitro. In vivo delivery of TK-p53-NTR genes by SP94-targeted PLGA-PEG-PEI NP in mice resulted in a strong expression of suicide genes selectively in tumors, and subsequent administration of GCV and CB1954 led to a decline in tumor growth, and established a superior therapeutic outcome against HCC. We demonstrate a highly efficient approach that exogenously supplements p53 to enable synergy with the outcome of TK-NTR suicide gene therapy against HCC.


Assuntos
Antineoplásicos , Carcinoma Hepatocelular/metabolismo , Terapia Genética/métodos , Nanopartículas/química , Polietilenoimina/química , Animais , Antineoplásicos/química , Antineoplásicos/metabolismo , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Feminino , Neoplasias Hepáticas/metabolismo , Camundongos , Camundongos Nus , Nitrorredutases/genética , Nitrorredutases/metabolismo , Poliésteres/química , Polietilenoglicóis/química , Pró-Fármacos/metabolismo , Pró-Fármacos/farmacologia , Timidina Quinase/genética , Timidina Quinase/metabolismo , Transfecção , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo
15.
Molecules ; 25(3)2020 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-32012733

RESUMO

The aim of this study was to develop a prodrug of ubiquinol-10 (UqH-10), the active form of ubiquinone-10 (Uq-10), for oral delivery. Bioavailability of UqH-10 is hampered by its high susceptibility to oxidation and water-insolubility. We prepared three novel N,N-dimethylglycine ester derivatives of UqH-10, including a 1-monoester (UqH-1-DMG), 4-monoester (UqH-4-DMG), and 1,4-bis-ester (UqH-DMG), and assessed their physicochemical properties in vitro and in vivo. UqH-DMG spontaneously formed an aqueous micelle solution comprising 20 nm particles at 36.5 °C. Cationic UqH-DMG formed nano-sized (5 nm) mixed-micelles with taurocholic acid. Reconversion of the derivatives to UqH-10 was accelerated in human liver microsomes. The oral bioavailability of UqH-10 after administration of UqH-derivatives or Uq-10 was determined in fasted and postprandial rats secreting normal and high levels of bile, respectively. In fasted rats, plasma UqH-10 after UqH-derivatives administration reached Cmax at 2-3 h and after Uq-10 administration, it remained low. The AUC0-24h of UqH-10 after UqH-derivatives administration was 2-3-fold higher than that after Uq-10 administration. In postprandial rats, the Tmax of UqH-10 after UqH-derivatives administration was an hour earlier than after Uq-10 administration. In conclusion, cationic UqH-derivatives are convenient prodrugs that enhance UqH-10 bioavailability by forming nanosized mixed-micelles with intestinal bile acids.


Assuntos
Ânions/química , Ácidos e Sais Biliares/química , Cátions/química , Absorção Intestinal/efeitos dos fármacos , Micelas , Pró-Fármacos/administração & dosagem , Ubiquinona/administração & dosagem , Administração Oral , Animais , Ânions/metabolismo , Ácidos e Sais Biliares/metabolismo , Disponibilidade Biológica , Transporte Biológico , Masculino , Nanopartículas , Oxirredução , Pró-Fármacos/química , Pró-Fármacos/metabolismo , Ratos , Ratos Sprague-Dawley , Ubiquinona/química , Ubiquinona/metabolismo
16.
PLoS One ; 15(1): e0227104, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31914458

RESUMO

Despite the availability of highly effective direct-acting antiviral (DAA) regimens for the treatment of hepatitis C virus (HCV) infections, sustained viral response (SVR) rates remain suboptimal for difficult-to-treat patient populations such as those with HCV genotype 3, cirrhosis or prior treatment experience, warranting development of more potent HCV replication antivirals. AT-527 is the hemi-sulfate salt of AT-511, a novel phosphoramidate prodrug of 2'-fluoro-2'-C-methylguanosine-5'-monophosphate that has potent in vitro activity against HCV. The EC50 of AT-511, determined using HCV laboratory strains and clinical isolates with genotypes 1-5, ranged from 5-28 nM. The active 5'-triphosphate metabolite, AT-9010, specifically inhibited the HCV RNA-dependent RNA polymerase. AT-511 did not inhibit the replication of other selected RNA or DNA viruses in vitro. AT-511 was approximately 10-fold more active than sofosbuvir (SOF) against a panel of laboratory strains and clinical isolates of HCV genotypes 1-5 and remained fully active against S282T resistance-associated variants, with up to 58-fold more potency than SOF. In vitro, AT-511 did not inhibit human DNA polymerases or elicit cytotoxicity or mitochondrial toxicity at concentrations up to 100 µM. Unlike the other potent guanosine analogs PSI-938 and PSI-661, no mutagenic O6-alkylguanine bases were formed when incubated with cytochrome P450 (CYP) 3A4, and AT-511 had IC50 values ≥25 µM against a panel of CYP enzymes. In hepatocytes from multiple species, the active triphosphate was the predominant metabolite produced from the prodrug, with a half-life of 10 h in human hepatocytes. When given orally to rats and monkeys, AT-527 preferentially delivered high levels of AT-9010 in the liver in vivo. These favorable preclinical attributes support the ongoing clinical development of AT-527 and suggest that, when used in combination with an HCV DAA from a different class, AT-527 may increase SVR rates, especially for difficult-to-treat patient populations, and could potentially shorten treatment duration for all patients.


Assuntos
Antivirais/farmacologia , Guanosina/farmacologia , Hepacivirus/efeitos dos fármacos , Hepatite C/tratamento farmacológico , Pró-Fármacos/farmacologia , Animais , Antivirais/química , Antivirais/metabolismo , Antivirais/farmacocinética , Linhagem Celular , Descoberta de Drogas , Avaliação Pré-Clínica de Medicamentos , Feminino , Guanosina/análogos & derivados , Guanosina/metabolismo , Guanosina/farmacocinética , Haplorrinos , Hepacivirus/genética , Hepatócitos/efeitos dos fármacos , Hepatócitos/metabolismo , Hepatócitos/virologia , Humanos , Masculino , Camundongos , Pró-Fármacos/química , Pró-Fármacos/metabolismo , Pró-Fármacos/farmacocinética , Ratos
18.
Nanoscale ; 12(4): 2793-2809, 2020 Jan 28.
Artigo em Inglês | MEDLINE | ID: mdl-31961354

RESUMO

In the field of nanomedicine, nanostructured nanoparticles (NPs) made of self-assembling prodrugs emerged in the recent years with promising properties. In particular, squalene-based drug nanoparticles have already shown their efficiency through in vivo experiments. However, a complete pattern of their stability and interactions in the blood stream is still lacking. In this work we assess the behavior of squalene-adenosine (SQAd) nanoparticles - whose neuroprotective effect has already been demonstrated in murine models - in the presence of fetal bovine serum (FBS) and of bovine serum albumin (BSA), the main protein of blood plasma. Extensive physicochemical characterizations were performed using Small Angle Neutron Scattering (SANS), cryogenic transmission electron microscopy (Cryo-TEM), circular dichroism (CD), steady-state fluorescence spectroscopy (SSFS) and isothermal titration calorimetry (ITC) as well as in silico by means of ensemble docking simulations with human serum albumin (HSA). Significant changes in the colloidal stability of the nanoparticles in the presence of serum albumin were observed. SANS, CD and SSFS analyses demonstrated an interaction between SQAd and BSA, with a partial disassembly of the nanoparticles in the presence of BSA and the formation of a complex between SQAd and BSA. The interaction free energy of SQAd nanoparticles with BSA derived from ITC experiments, is about -8 kcal mol-1 which is further supported in silico by ensemble docking simulations. Overall, our results show that serum albumin partially disassembles SQAd nanoparticles by extracting individual SQAd monomers from them. As a consequence, the SQAd nanoparticles would act as a circulating reservoir in the blood stream. The approach developed in this study could be extended to other soft organic nanoparticles.


Assuntos
Adenosina/química , Nanopartículas/química , Albumina Sérica/metabolismo , Esqualeno/química , Adenosina/metabolismo , Animais , Sítios de Ligação , Coloides , Estabilidade de Medicamentos , Humanos , Camundongos , Nanopartículas/metabolismo , Pró-Fármacos/química , Pró-Fármacos/metabolismo , Ligação Proteica , Albumina Sérica/química , Esqualeno/metabolismo
19.
Colloids Surf B Biointerfaces ; 188: 110739, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31901623

RESUMO

This study aims to develop in situ microemulsion-gel (ME-Gel) obtained from hydroxypropyl methylcellulose (HPMC) films for transdermal administration of Zidovudine (AZT). Firstly, HPMC films containing propylene glycol (PG) and eucalyptus oil (EO) were obtained and characterized. Later, a pseudo-ternary phase diagram composed of water, EO, tween 80 and PG was obtained and one microemulsion (ME) with a similar proportion of the film components was obtained. ME was transformed in ME-Gel by the incorporation of HPMC. Finally, HPMC films were hydrated with Tween 80 solution to yield in situ ME-Gel and its effect on AZT skin permeation was compared with HPMC film hydrated with water (F5hyd). The results showed that the ME and ME-Gel presented a droplet size of 16.79 and 122.13 µm, respectively, polydispersity index (PDI) < 0.39 and pH between 5.10 and 5.40. The incorporation of HPMC resulted in viscosity about 2 times higher than the use of ME. The presence of AZT did not alter the formulation properties. The in situ ME-Gel promoted a two-fold increase in the permeated amount of AZT compared to F5hyd. The results suggest that it was possible to obtain an ME-Gel in situ from HPMC films and that its effect on transdermal permeation of AZT was significant.


Assuntos
Metilcelulose/química , Pró-Fármacos/química , Zidovudina/química , Administração Cutânea , Animais , Emulsões/administração & dosagem , Emulsões/química , Emulsões/metabolismo , Óleo de Eucalipto/administração & dosagem , Óleo de Eucalipto/química , Óleo de Eucalipto/metabolismo , Géis/administração & dosagem , Géis/química , Géis/metabolismo , Masculino , Metilcelulose/administração & dosagem , Metilcelulose/metabolismo , Tamanho da Partícula , Pró-Fármacos/administração & dosagem , Pró-Fármacos/metabolismo , Propilenoglicol/administração & dosagem , Propilenoglicol/química , Propilenoglicol/metabolismo , Ratos , Ratos Wistar , Pele/química , Pele/metabolismo , Absorção Cutânea , Propriedades de Superfície , Zidovudina/administração & dosagem , Zidovudina/metabolismo
20.
Adv Mater ; 32(4): e1904011, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31793717

RESUMO

Biodegradable nanoprodrugs, inheriting the antitumor effects of chemotherapy drugs and overcoming the inevitable drawback of side effects on normal tissues, hold promise as next-generation cancer therapy candidates. Biodegradable nanoprodrugs of transferrin-modified MgO2 nanosheets are developed to selectively deliver reactive oxygen species to cancer cells for molecular dynamic therapy strategy. The nanosheets favor the acidic and low catalase activity tumor microenvironment to react with proton and release nontoxic Mg2+ . This reaction simultaneously produces abundant H2 O2 to induce cell death and damage the structure of transferrin to release Fe3+ , which will react with H2 O2 to produce highly toxic ·OH to kill tumor cells.


Assuntos
Nanopartículas/química , Espécies Reativas de Oxigênio/metabolismo , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Humanos , Peróxido de Hidrogênio/metabolismo , Peróxido de Hidrogênio/uso terapêutico , Peróxido de Hidrogênio/toxicidade , Óxido de Magnésio/química , Neoplasias/tratamento farmacológico , Neoplasias/patologia , Pró-Fármacos/metabolismo , Pró-Fármacos/uso terapêutico , Pró-Fármacos/toxicidade , Espécies Reativas de Oxigênio/uso terapêutico , Espécies Reativas de Oxigênio/toxicidade , Transferrinas/química
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