RESUMO
Ferroptosis and inflammation induced by cerebral hemorrhage result in an excessive inflammatory response and irreversible neuronal injury. Alleviating ferroptosis might be an effective way to prevent neuroinflammatory injury and promote neural functional recovery. Pyridoxal isonicotinoyl hydrazine (PIH), a lipophilic iron-chelating agent, has been reported to reduce excess iron-induced cytotoxicity. However, whether PIH could ameliorate the effects of hemorrhagic stroke is not completely understood. In the present study, the preventive effects of PIH in an intracerebral hemorrhage (ICH) mouse model were investigated. Neurological score, rotarod test, and immunofluorescence around the hematoma were assessed to evaluate the effects of PIH on hemorrhagic injury. The involvement of ferroptosis and inflammation was also examined in vitro to explore the underlying mechanism. Results showed that administration of PIH prevented neuronal cell death and reduced lipid peroxidation in Erastin-treated PC-12 cells. In vivo, mice treated with PIH after ICH attenuated neurological deficit scores. Additionally, we found PIH reduced ROS production, iron accumulation, and lipid peroxidation around the hematoma peripheral tissue. Meanwhile, ICH mice treated with PIH showed an upregulation of the key ferroptosis enzyme, glutathione peroxidase 4, and downregulation of cyclooxygenase-2. Moreover, PIH administration inhibited proinflammatory polarization and reduced interleukin-1 beta and tumor necrosis factor alpha in ICH mice. Collectively, these results demonstrated that PIH protects mice against hemorrhage stroke, which was associated with mitigation of inflammation and ferroptosis.
Assuntos
Hemorragia Cerebral/tratamento farmacológico , Ferroptose/efeitos dos fármacos , Isoniazida/análogos & derivados , Piridoxal/análogos & derivados , Animais , Apoptose/efeitos dos fármacos , Morte Celular/efeitos dos fármacos , Hemorragia Cerebral/metabolismo , Compostos Férricos/farmacologia , Ferroptose/fisiologia , Inflamação/tratamento farmacológico , Inflamação/prevenção & controle , Ferro/metabolismo , Quelantes de Ferro/farmacologia , Isoniazida/metabolismo , Isoniazida/farmacologia , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Neurônios/metabolismo , Piridoxal/metabolismo , Piridoxal/farmacologiaRESUMO
In this work we report on the antitumor properties of a series of pincer-type metallocomplexes [Hg2(HL-keto)Cl4]n (1), [Hg(HL-keto)I2] (2) and [Mn(HL-zwitterion)Cl2]âMeOH (3âMeOH), derived from N'-(1-(pyridin-2-yl)ethylidene)isonicotinohydrazide (HL) and corresponding metal salts. The Hg(II) and Mn(II) salts are chelated by the keto (HL-keto) or zwitterionic (HL-zwitterion) form of HL, respectively. The cytotoxic effects of these compounds have been accessed against lung adenocarcinoma (A549) and hepatocellular carcinoma (HepG2 and Huh7) cell lines. Complexes 1 and 2 were found to be most efficient against the cell line Huh7 with IC50 value of 2.56 and 9.90 µM, respectively, while they exhibit moderate activity towards cell lines A549 and HepG2, as evidenced from IC50 values in the range 27.98-56.99 µM. Complex 3âMeOH is less efficient towards all the three cell lines with relatively high IC50 values. The mechanisms of the metallocomplexes killing the aforementioned cells were elucidated by flow cytometry, colony formation and polymerase chain reaction (PCR) analysis of apoptosis related expression of the genes. The results of the cytotoxic effects and antitumor activity on different cell lines are affected by the metal nature and the presence of the coordinated halide.
Assuntos
Antineoplásicos/farmacologia , Complexos de Coordenação/farmacologia , Isoniazida/análogos & derivados , Isoniazida/farmacologia , Antineoplásicos/síntese química , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Complexos de Coordenação/síntese química , Inibidor de Quinase Dependente de Ciclina p21/metabolismo , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Estrutura Molecular , Proteína Supressora de Tumor p53/metabolismoRESUMO
NAD+ is a critical molecule that is involved in multiple cellular functions. CD38 is a multifunctional enzyme with NAD+ nucleosidase activity. Our previous work revealed the CD38-dependent interactions of isoniazid (INH), an antituberculosis drug, with NAD+ to form INH-NAD adduct. In the current work, our metabolomic analysis discovered a novel NAD+ adduct with acetylisoniazid (AcINH), a primary INH metabolite mediated by N-acetyltransferase (NAT), and we named it AcINH-NAD. Using Nat1/2(-/-) and Cd38(-/-) mice, we determined that AcINH-NAD formation is dependent on both NAT and CD38. Because NAT is expressed in hepatocytes (HP), whereas CD38 is expressed in Kupffer cells (KC) and hepatic stellate cells (HSC), we explored cell type-specific roles of CD38 in the formation of AcINH-NAD as well as INH-NAD. We found that both INH-NAD and AcINH-NAD were produced in the incubation of INH or AcINH with KC and HSC but not in HP. These data suggest that hepatic nonparenchymal cells, such as KC and HSC, are the major cell types responsible for the CD38-dependent interactions of INH with NAD+ in the liver. SIGNIFICANCE STATEMENT: The current study identified AcINH-NAD as a novel metabolite of INH in the liver. Our work also revealed the essential roles of nonparenchymal cells, including Kupffer cells and hepatic stellate cells, in the CD38-dependent interactions of NAD+ with INH, leading to the formation of both INH-NAD and AcINH-NAD in the liver. These data can be used to guide the future studies on the mechanisms of INH and NAD+ interactions and their contributions to INH-induced liver injury.
Assuntos
ADP-Ribosil Ciclase 1/metabolismo , Antituberculosos/farmacocinética , Isoniazida/análogos & derivados , Fígado/metabolismo , NAD/metabolismo , Animais , Arilamina N-Acetiltransferase/genética , Arilamina N-Acetiltransferase/metabolismo , Células Cultivadas , Células Estreladas do Fígado/metabolismo , Isoenzimas/genética , Isoniazida/farmacocinética , Células de Kupffer/metabolismo , Fígado/citologia , Masculino , Camundongos , Camundongos Knockout , Modelos Animais , Cultura Primária de Células , SuínosRESUMO
Background: A major focus of tuberculosis drug discovery is aimed at the development of novel antibiotics with activity against drug-resistant strains of Mycobacterium tuberculosis. Results: We have synthesized ten isoniazid derivatives and investigated for antibacterial activity toward M. tuberculosis H37Rv and isoniazid-resistant strain SRI 1369. It was revealed that only one compound, isonicotinic acid (1-methyl-1H-pyrrol-2-ylmethylene)-hydrazide (1), is active toward isoniazid-resistant strain with minimum inhibitory concentration value of 0.14 µM. This compound is not cytotoxic toward human liver cells (HepG2; IC50 >100 µM), demonstrates good permeability in Caco-2 cells. Accordingly to the results of plasma protein binding assay, unbound fraction of compound 1, which potentially exhibits pharmacologic effects, is 57.9%. Conclusion: Therefore, isonicotinic acid (1-methyl-1H-pyrrol-2-ylmethylene)-hydrazide is a promising compound for further preclinical studies.
Assuntos
Antituberculosos/antagonistas & inibidores , Antituberculosos/farmacologia , Isoniazida/análogos & derivados , Isoniazida/farmacologia , Mycobacterium tuberculosis/efeitos dos fármacos , Sistema Enzimático do Citocromo P-450/efeitos dos fármacos , Humanos , Ácidos Isonicotínicos/química , Macrófagos , Testes de Sensibilidade Microbiana , Tuberculose/tratamento farmacológicoRESUMO
We studied the response of the extracellular matrix of the lungs and liver in mice with BCGinduced granulomatosis (3 months) after inhalation and intraperitoneal administration of liposome-encapsulated dextrazide (LEDZ): a conjugate of oxidized dextran (40 kDa) and isonicotinic acid hydrazide (INH). LEDZ inhalation proved to be more effective in reducing fibrosis severity, both in the lungs and liver. However, the mechanisms of the antifibrotic effect were different: increased degradation and reduced collagen synthesis in the lungs and reduced collagen synthesis and collagen degradation in the liver. This suggest that drug administration routes and delivery to the target organs are crucially important in the therapy of tuberculosis. The antifibrotic effect depended on LEDZ administration route and was more potent after LEDZ inhalation.
Assuntos
Antituberculosos/administração & dosagem , Granuloma do Sistema Respiratório/tratamento farmacológico , Lipossomos/administração & dosagem , Fibrose Pulmonar/tratamento farmacológico , Animais , Antituberculosos/química , Vacina BCG/efeitos adversos , Dextranos/administração & dosagem , Dextranos/química , Composição de Medicamentos , Granuloma do Sistema Respiratório/etiologia , Isoniazida/administração & dosagem , Isoniazida/análogos & derivados , Isoniazida/química , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Oxirredução , Fibrose Pulmonar/etiologia , Tuberculose Pulmonar/complicações , Tuberculose Pulmonar/prevenção & controleRESUMO
The study examined effectiveness of liposomal form of dextrazide (inhaled or intraperitoneal), free dextrazide (intraperitoneal), and isoniazid (intraperitoneal) in the treatment of BALB/c mice with BCG-induced granulomatosis. The mice were infected with mycobacteria tuberculosis 3 months prior to onset of treatment. The preparations under examinations were administered twice a week over 2 months. The decrease of the number and size of macrophagal granulomas in mice BCG-induced granulomatosis during treatment was determined by the number of living mycobacteria tuberculosis in these granulomas. The most effective treatment was achieved with liposomal form of dextrazide (a conjugate of oxidized dextran with isonicotinic acid hydrazide). Macrophages with captured mycobacteria tuberculosis, dextrazide, and dextrazide-loaded liposomes can be incorporated into granulomas. The antimycobacterial effect of dextrazide is an important factor preventing the destructive processes in granulomas and organs via a decrease in the prodestructive potential of lysosomes in macrophages realized after their migration from granulomas.
Assuntos
Antituberculosos/uso terapêutico , Vacina BCG/efeitos adversos , Dextranos/uso terapêutico , Granuloma do Sistema Respiratório , Isoniazida/uso terapêutico , Pulmão/microbiologia , Tuberculose Pulmonar/tratamento farmacológico , Animais , Antituberculosos/química , Dextranos/química , Combinação de Medicamentos , Granuloma do Sistema Respiratório/induzido quimicamente , Granuloma do Sistema Respiratório/tratamento farmacológico , Granuloma do Sistema Respiratório/etiologia , Granuloma do Sistema Respiratório/microbiologia , Isoniazida/análogos & derivados , Isoniazida/química , Pulmão/efeitos dos fármacos , Pulmão/patologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Mycobacterium tuberculosis/efeitos dos fármacos , Mycobacterium tuberculosis/isolamento & purificação , Mycobacterium tuberculosis/fisiologia , Resultado do Tratamento , Tuberculose Pulmonar/microbiologia , Tuberculose Pulmonar/patologia , Tuberculose Pulmonar/prevenção & controleRESUMO
Monitoring hypochlorite anion (ClO-) in living cells is particularly meaningful and valuable, because over-exposure of the ClO- may cause a potential health hazard towards animals and humans. Considering the special structure and properties of the gemini surfactant, a novel amphiphilic gemini-iridium complex Ir[(ppy-iso)2(bpy-tma2Br2)] (Ir-iso) with isoniazide as a recognition site for ClO- was designed. The Ir-iso possessed an excellent water-solubility as well as a strong ClO- binding capacity, as revealed from the rapid response of emission signal towards ClO-. It was worth noting that such probe had a highly-specific selectivity with a low detection limit (20.5â¯nM) and was suitable in physiological environment. The cell viability assay, cell imaging, and co-location studies further proved that the Ir-iso had little cytotoxicity and was specifically localized in the mitochondria of breast cancer cells, being a promising candidate of chemo-sensor to detect the endogenous ClO- in living cells.
Assuntos
Complexos de Coordenação/química , Ácido Hipocloroso/análise , Isoniazida/análogos & derivados , Substâncias Luminescentes/química , Mitocôndrias/metabolismo , Animais , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Complexos de Coordenação/síntese química , Complexos de Coordenação/toxicidade , Irídio/química , Isoniazida/síntese química , Isoniazida/toxicidade , Limite de Detecção , Substâncias Luminescentes/síntese química , Substâncias Luminescentes/toxicidade , Medições Luminescentes/métodos , Camundongos , Microscopia Confocal/métodosRESUMO
Bacteria belonging to Mycobacterium avium complex are organisms of low pathogenicity that infect immunosuppressed individuals. Infection is treated with an antimicrobial macrolide, Clarithromycin (CAM) or Azitromycin, associated with Ethambutol and Rifabutin during 12 months. Regimen long duration and side effects hinder patient's commitment to treatment favoring emergence of antibiotic resistance. In this present study, we evaluated the activity of JVA, an Isoniazid (INH) derivative, against M. avium 2447, a clinical isolate. We demonstrated that JVA reduces M. avium 2447 growth in macrophages, more efficiently than CAM and INH. In order to explore JVA mechanism of action, we investigated compound properties and performed pH-dependent stability studies. Our results suggest an enhanced ability of JVA to cross biological membranes. Furthermore, we suggest that in acidic conditions of macrophages' phagosomes, where mycobacteria replicate, JVA would be promptly hydrolyzed to INH, delivering the adduct INH-nicotinamide adenine dinucleotide and thus inhibiting M. avium 2447 growth.
Assuntos
Antituberculosos/farmacologia , Isoniazida/análogos & derivados , Complexo Mycobacterium avium/efeitos dos fármacos , Infecção por Mycobacterium avium-intracellulare/tratamento farmacológico , Análise de Variância , Animais , Hidrazonas/farmacologia , Macrófagos/microbiologia , Camundongos Endogâmicos C57BL , Testes de Sensibilidade Microbiana , Complexo Mycobacterium avium/crescimento & desenvolvimentoRESUMO
The formation of isonicotinyl-nicotinamide adenine dinucleotide (INH-NAD+) by the mycobacterial catalase-peroxidase enzyme, KatG, was known to be the major component of the mode of action of isoniazid (INH), an anti-tuberculosis drug. However, there are other enzymes that may catalyze this reaction. We have previously reported that neutrophil myeloperoxidase (MPO) is capable of metabolizing INH through the formation of INH-NAD+ adduct, which could be attributed to being a possible mode of action of INH. However, eosinophilic infiltration of the lungs is more pronounced and characteristic of granulomas in Mycobacterium tuberculosis-infected patients. Thus, the aim of the present study is to investigate the role of eosinophil peroxidase (EPO), a key eosinophil enzyme, during INH metabolism and the formation of its active metabolite, INH-NAD+ using purified EPO and eosinophils isolated from asthmatic donors. UV-Vis spectroscopy revealed INH oxidation by EPO led to a new product (λmaxâ¯=â¯326â¯nm) in the presence of NAD+. This adduct was confirmed to be INH-NAD+ using LC-MS analysis where the intact adduct was detected (m/zâ¯=â¯769). Furthermore, EPO catalyzed the oxidation of INH and formed several free radical intermediates as assessed by electron paramagnetic resonance (EPR) spin-trapping; a carbon-centred radical, which is considered to be the reactive metabolite that binds with NAD+, was found when superoxide dismutase was included in the reaction. Our findings suggest that eosinophilic EPO may also play a role in the pharmacological activity of INH through the formation of INH-NAD+ adduct, and supports further evidence that human cells and enzymes are capable of producing the active metabolite involved in tuberculosis treatment.
Assuntos
Peroxidase de Eosinófilo/metabolismo , Eosinófilos/enzimologia , Isoniazida/análogos & derivados , Isoniazida/metabolismo , NAD/análogos & derivados , NAD/metabolismo , Asma/metabolismo , Asma/patologia , Cromatografia Líquida de Alta Pressão , Espectroscopia de Ressonância de Spin Eletrônica , Eosinófilos/química , Eosinófilos/efeitos dos fármacos , Humanos , Isoniazida/sangue , Isoniazida/química , Isoniazida/farmacologia , Espectrometria de Massas , Mycobacterium tuberculosis/efeitos dos fármacos , Mycobacterium tuberculosis/patogenicidade , NAD/sangue , NAD/química , Oxirredução , Fator de Ativação de Plaquetas/farmacologia , Superóxido Dismutase/metabolismoRESUMO
In a mouse model, rifampicin and isoniazid combination treatment results in cholestatic liver injury that is associated with an increase in protoporphyrin IX, the penultimate heme precursor. Both ferrochelatase (FECH/Fech) and aminolevulinic acid synthase 1 (ALAS1/Alas1) are crucial enzymes in regulating heme biosynthesis. Isoniazid has recently been reported to upregulate Alas1 but downregulate Fech protein levels in mice; however, the mechanism by which isoniazid mediates disruption of heme synthesis has been unclear. Two metabolites of isoniazid, pyridoxal isonicotinoyl hydrazone (PIH, the isoniazid-vitamin B6 conjugate) and hydrazine, have been detected in the urine of humans treated with isoniazid. Here we show that, in primary human hepatocytes and the human hepatocellular carcinoma cell line HepG2/C3A, (1) isoniazid treatment increases Alas1 protein levels but decreases Fech levels; (2) hydrazine treatment upregulates Alas1 protein and Alas1 mRNA levels; (3) PIH treatment decreases Fech protein levels, but not Fech mRNA levels; and (4) PIH is detected after isoniazid treatment, with levels increasing further when exogenous vitamin B6 analogs are coadministered. In addition, the PIH-mediated downregulation of human FECH is associated with iron chelation. Together, these data demonstrate that hydrazine upregulates ALAS1, whereas PIH downregulates FECH, suggesting that the metabolites of isoniazid mediate its disruption of heme biosynthesis by contributing to protoporphyrin IX accumulation.
Assuntos
5-Aminolevulinato Sintetase/metabolismo , Heme/biossíntese , Hidrazinas/farmacologia , Isoniazida/análogos & derivados , Isoniazida/metabolismo , Isoniazida/farmacologia , Piridoxal/análogos & derivados , Animais , Carcinoma Hepatocelular , Ferroquelatase/metabolismo , Células Hep G2 , Hepatócitos/metabolismo , Humanos , Ferro/fisiologia , Fígado/metabolismo , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Protoporfirinas/metabolismo , Piridoxal/farmacologia , Rifampina/metabolismo , Rifampina/farmacologia , Vitamina B 6/metabolismoRESUMO
The development of versatile antitumor agents with tumor-imaging, targeting and therapeutic activity is promising for clinical cancer therapy. Prostate cancer is still the one of the leading threats to males. Current therapies have restricted clinical efficiency for patients with advanced and metastatic prostate cancer. Recent studies demonstrate that monoamine oxidase A (MAOA) levels elevate with prostate cancer aggression and metastasis. In addition, MAOA inhibitor therapies have been reported as an effective means to reduce the metastasis of prostate cancer and extend mouse survival. Thus, these findings provide evidence that MAOA is promising for the treatment of metastatic and advanced prostate cancer. Herein, three isoniazid (INH)-dye conjugates were synthesized by conjugating MAOA inhibitor INH with mitochondria-targeting NIRF heptamethine dyes to improve the therapeutic efficacy of prostate cancer. These INH-dye conjugates could accumulate in PC-3 cellular mitochondria via organic anion transport peptide (OATP), increase ROS generation, and induce cancer cells apoptosis. In prostate cancer bearing xenografts, INH-dye conjugates showed significantly improved tumor-homing characteristics, resulting in potent antitumor activity via a reduction in MAOA activity. These results suggest that INH-dye conjugates have great potential to be used as versatile antitumor agents with prostate cancer targeting, NIR imaging, and potent antitumor efficacy.
Assuntos
Antituberculosos/química , Antituberculosos/uso terapêutico , Isoniazida/análogos & derivados , Isoniazida/uso terapêutico , Inibidores da Monoaminoxidase/química , Inibidores da Monoaminoxidase/uso terapêutico , Neoplasias da Próstata/tratamento farmacológico , Animais , Antituberculosos/farmacologia , Corantes/química , Corantes/farmacologia , Corantes/uso terapêutico , Reposicionamento de Medicamentos/métodos , Humanos , Isoniazida/farmacologia , Masculino , Camundongos Endogâmicos BALB C , Camundongos Nus , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Mitocôndrias/patologia , Monoaminoxidase/metabolismo , Inibidores da Monoaminoxidase/farmacologia , Células PC-3 , Neoplasias da Próstata/metabolismo , Neoplasias da Próstata/patologiaRESUMO
The development of novel drugs is essential for the treatment of tuberculosis and other mycobacterial infections in future. A series of N-alkyl-2-isonicotinoylhydrazine-1-carboxamides was synthesized from isoniazid (INH) and then cyclized to N-alkyl-5-(pyridin-4-yl)-1,3,4-oxadiazole-2-amines. All derivatives were characterised spectroscopically. The compounds were screened for their in vitro antimycobacterial activity against susceptible and multidrug-resistant Mycobacterium tuberculosis (Mtb.) and nontuberculous mycobacteria (NTM; M. avium, M. kansasii). The most active carboxamides were substituted by a short n-alkyl, their activity was comparable to INH with minimum inhibitory concentrations (MICs) against Mtb. of 0.5-2⯵M. Moreover, they are non-toxic for HepG2, and some of them are highly active against INH-resistant NTM (MICs ≥4⯵M). Their cyclization to 1,3,4-oxadiazoles did not increase the activity. The experimentally proved mechanism of action of 2-isonicotinoylhydrazine-1-carboxamides consists of the inhibition of enoyl-ACP reductase (InhA) in a way similar to INH, which is blocking the biosynthesis of mycolic acids. N-Dodecyl-5-(pyridin-4-yl)-1,3,4-oxadiazol-2-amine as the most efficacious oxadiazole inhibits growth of both susceptible and drug-resistant Mtb. strains with uniform MIC values of 4-8⯵M with no cross-resistance to antitubercular drugs including INH. The mechanism of action is not elucidated but it is different from INH. Obtained results qualify these promising derivatives for further investigation.
Assuntos
Antituberculosos/química , Antituberculosos/farmacologia , Isoniazida/análogos & derivados , Isoniazida/farmacologia , Mycobacterium tuberculosis/efeitos dos fármacos , Oxidiazóis/química , Oxidiazóis/farmacologia , Antituberculosos/síntese química , Farmacorresistência Bacteriana , Células Hep G2 , Humanos , Isoniazida/síntese química , Testes de Sensibilidade Microbiana , Oxidiazóis/síntese química , Tuberculose/tratamento farmacológico , Tuberculose/microbiologiaRESUMO
The interaction of pyridoxal isonicotinoyl hydrazone (PIH) and salicylaldehyde isonicotinoyl hydrazone (SIH), two important biologically active chelators, with iron has been investigated by spectrophotometric methods. High iron(III) affinity constants were determined for PIH, logß2=37.0 and SIH, logß2=37.6. The associated redox potentials of the iron complexes were determined using cyclic voltammetry at pH7.4 as +130mV (vs normal hydrogen electrode, NHE) for PIH and +136mV(vs NHE) for SIH. These redox potentials are much higher than those corresponding to iron chelators in clinical use, namely deferiprone, -620mV; desferasirox, -600mV and desferrioxamine, -468mV. Although the positive redox potentials of SIH and PIH are similar to that of EDTA, namely +120mV, the iron complexes of these two hydrazone chelators, unlike the iron complex of EDTA, do not redox cycle in the presence of vitamin C. These properties render PIH and SIH as excellent scavengers of iron, under biological conditions. Both SIH and PIH scavenge mononuclear iron(II) and iron(III) rapidly. These fast kinetic properties of the hydrazone-based chelators provide a ready explanation for the adoption of SIH in fluorescence-based methods for the quantification of cytosolic iron(II).
Assuntos
Aldeídos/química , Compostos Férricos/química , Compostos Ferrosos/química , Hidrazonas/química , Isoniazida/análogos & derivados , Piridoxal/análogos & derivados , Ácido Ascórbico/química , Ácido Edético/química , Técnicas Eletroquímicas/métodos , Eletrodos , Concentração de Íons de Hidrogênio , Quelantes de Ferro/química , Isoniazida/química , Cinética , Oxirredução , Piridoxal/químicaRESUMO
The emergence of multidrug-resistant strains of Mycobacterium tuberculosis (MTB) represents a major threat to global health. Isoniazid (INH) is a prodrug used in the first-line treatment of tuberculosis. It undergoes oxidation by a catalase-peroxidase KatG, leading to generation of an isonicotinoyl radical that reacts with NAD(H) forming the INH-NADH adduct as the active metabolite. A redox-mediated activation of isoniazid using an iron metal complex was previously proposed as a strategy to overcome isoniazid resistance due to KatG mutations. Here, we have prepared a series of iron metal complexes with isoniazid and analogues, containing alkyl substituents at the hydrazide moiety, and also with pyrazinamide derivatives. These complexes were activated by H2O2 and studied by ESR and LC-MS. For the first time, the formation of the oxidized INH-NAD adduct from the pentacyano(isoniazid)ferrate(II) complex was detected by LC-MS, supporting a redox-mediated activation, for which a mechanistic proposition is reported. ESR data showed all alkylated hydrazides, in contrast to non-substituted hydrazides, only generated alkyl-based radicals. The structural modifications did not improve minimal inhibitory concentration (MIC) against MTB in comparison to isoniazid iron complex, providing support to isonicotinoyl radical formation as a requirement for activity. Nonetheless, the pyrazinoic acid hydrazide iron complex showed redox-mediated activation using H2O2 with generation of a pyrazinoyl radical intermediate and production of pyrazinoic acid, which is in fact the active metabolite of pyrazinamide prodrug. Thereby, this strategy can also unveil new opportunities for activation of this type of drug.
Assuntos
Antituberculosos/farmacologia , Complexos de Coordenação/farmacologia , Compostos Ferrosos/farmacologia , Isoniazida/análogos & derivados , Isoniazida/farmacologia , Antituberculosos/síntese química , Antituberculosos/química , Complexos de Coordenação/síntese química , Complexos de Coordenação/química , Compostos Ferrosos/síntese química , Compostos Ferrosos/química , Isoniazida/síntese química , Isoniazida/química , Testes de Sensibilidade Microbiana , Modelos Químicos , Mycobacterium tuberculosis/efeitos dos fármacos , OxirreduçãoRESUMO
Cancer is ultimately the result of cells that hysterically grow and do not die. Cells can experience uncontrolled growth if there are mutations to DNA, and therefore, alterations to the genes involved in cell division. Cancer occurs when a cell's gene mutations make the cell unable to correct DNA damage and is unable to destroy itself. There are over 100 different types of cancer each classified by the type of initially affected cell. Isoniazid, a well-known antitubercular agent has been reported to exhibit some cytotoxic activity. This finding prompt us to carry out this study where isoniazid and its sixteen derivatives were studied for any possible cytotoxic activity against Human astrocytoma SNB-19 cells, human Dukes' type C colorectal adenocarcinoma HCT-15 cells, human Dukes' type D colorectal adenocarcinoma COLO-205 cells, and human prostate adenocarcinoma (grade IV) PC-3 cells. Among the test compounds, SN-07 (a phenacyl derivative with para phenyl substitution) demonstrated slight cytotoxic effects on two types of human colorectal adenocarcinoma cells HCT-15 and COLO-205. Moreover, the acute toxicity of the compounds was also estimated in which some compounds were evaluated with more LD50 values than isoniazid.
Assuntos
Adenocarcinoma/tratamento farmacológico , Antineoplásicos/farmacologia , Astrocitoma/tratamento farmacológico , Neoplasias Encefálicas/tratamento farmacológico , Neoplasias Colorretais/tratamento farmacológico , Isoniazida/farmacologia , Neoplasias da Próstata/tratamento farmacológico , Adenocarcinoma/patologia , Animais , Antineoplásicos/toxicidade , Astrocitoma/patologia , Neoplasias Encefálicas/patologia , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Neoplasias Colorretais/patologia , Relação Dose-Resposta a Droga , Humanos , Concentração Inibidora 50 , Isoniazida/análogos & derivados , Isoniazida/toxicidade , Dose Letal Mediana , Masculino , Camundongos , Gradação de Tumores , Neoplasias da Próstata/patologia , Testes de Toxicidade Aguda/métodosRESUMO
The frontline tuberculosis (Tb) antibiotic isoniazid has been repurposed using a three component complex aimed at increasing the delivery efficiency and adding new avenues to its mechanism of action. This study focuses on pharmacokinetic studies of the isoniazid-sucrose-copper (II)-PEG-3350 complex. The assays include the Plasma Protein Binding Assay (85.8%), Caco-2 Permeability Assay (BâAPapp, 0.13 × 10-6 cm/s), Cytochrome P450 Inhibition Assay (i.e. CYP2B6, IC50 = 7.26 µM), In vitro microsomal Stability Assay (t1/2 NADPH-Dependent > 240 min), and HepG2 Cytotoxicity (no toxicity). The National Cancer Institute's 60 cell line panel is used to measure activity against cancer cells. The percent growth values averaged over all 60 cell lines indicates the complex has no anti-cancer activity, which also suggests a lack of general toxicity. It also provides data for the complexes specificity against Mycobacterium tuberculosis.
Assuntos
Antituberculosos/farmacocinética , Complexos de Coordenação/farmacocinética , Cobre/química , Inibidores do Citocromo P-450 CYP2B6/farmacocinética , Mucosa Intestinal/metabolismo , Isoniazida/farmacocinética , Mycobacterium tuberculosis/efeitos dos fármacos , Antituberculosos/química , Antituberculosos/toxicidade , Células CACO-2 , Sobrevivência Celular/efeitos dos fármacos , Complexos de Coordenação/toxicidade , Inibidores do Citocromo P-450 CYP2B6/química , Inibidores do Citocromo P-450 CYP2B6/toxicidade , Composição de Medicamentos , Meia-Vida , Células Hep G2 , Humanos , Absorção Intestinal , Isoniazida/análogos & derivados , Isoniazida/química , Isoniazida/toxicidade , PermeabilidadeRESUMO
Hydrazone complexes of Cu(II), Co(II), Zn(II), Ni(II) and Pt(II) with N-isonicotinoyl-N'-(3-metoxy-2 hydroxybenzaldehyde)-hydrazone (HL) were synthesized and characterized by different physico-chemical techniques including elemental and thermal analysis, magnetic susceptibility measurements, molar electric conductivity, as well as IR (infrared), ¹H-NMR and 13C-NMR (hydrogen and carbon nuclear magnetic resonance, UV-Vis (ultraviolet-visible), FAB (fast atom bombardment), EPR (electron paramagnetic resonance), and mass spectroscopies. The crystal structure of ligand was determined by single crystal X-ray diffraction studies. Spectral data showed that hydrazone behaves as an ONO tridentate ligand through the azomethine nitrogen, phenolate and keto oxygen atoms. For the copper(II) complexes, metal-ligand bonding parameters were evaluated from the EPR spectra. These parameters indicate the presence of in-plane π bonding. In addition, the f values of complexes 1-4 indicate small distortion from planarity. The effect of these complexes on proliferation of human breast cancer (MCF-7 and SKBR-3), human melanoma (A375), lung adenocarcinoma cells (NCI-H1573) and their antibacterial activity against Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus and Candida albicans strains were studied and compared with those of free ligand. The ligand and complexes 1-3 showed significant antimicrobial activity against the Gram-positive bacteria Staphylococcus aureus and Candida albicans in comparison to the control drugs. The complexes 2-4 could be potential antitumor agents, leading to a significant improvement of the cytotoxic activity when compared with HL.
Assuntos
Anti-Infecciosos/síntese química , Anti-Infecciosos/farmacologia , Antineoplásicos/síntese química , Antineoplásicos/farmacologia , Isoniazida/síntese química , Isoniazida/farmacologia , Metais/química , Bactérias/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Cobalto/química , Cobre/química , Fungos/efeitos dos fármacos , Humanos , Isoniazida/análogos & derivados , Testes de Sensibilidade Microbiana , Modelos Moleculares , Estrutura Molecular , Níquel/química , Platina/química , Solubilidade , Análise Espectral/métodos , Zinco/químicaRESUMO
The formation of isonicotinyl-nicotinamide adenine dinucleotide (INH-NAD(+)) via the mycobacterial catalase-peroxidase enzyme, KatG, has been described as the major component of the mode of action of isoniazid (INH). However, there are numerous human peroxidases that may catalyze this reaction. The role of neutrophil myeloperoxidase (MPO) in INH-NAD(+) adduct formation has never been explored; this is important, as neutrophils are recruited at the site of tuberculosis infection (granuloma) through infected macrophages' cell death signals. In our studies, we showed that neutrophil MPO is capable of INH metabolism using electron paramagnetic resonance (EPR) spin-trapping and UV-Vis spectroscopy. MPO or activated human neutrophils (by phorbol myristate acetate) catalyzed the oxidation of INH and formed several free radical intermediates; the inclusion of superoxide dismutase revealed a carbon-centered radical which is considered to be the reactive metabolite that binds with NAD(+). Other human metabolites, including N-acetyl-INH, N-acetylhydrazine, and hydrazine did not show formation of carbon-centered radicals, and either produced no detectable free radicals, N-centered free radicals, or superoxide, respectively. A comparison of these free radical products indicated that only the carbon-centered radical from INH is reducing in nature, based on UV-Vis measurement of nitroblue tetrazolium reduction. Furthermore, only INH oxidation by MPO led to a new product (λmax=326nm) in the presence of NAD(+). This adduct was confirmed to be isonicotinyl-NAD(+) using LC-MS analysis where the intact adduct was detected (m/z=769). The findings of this study suggest that neutrophil MPO may also play a role in INH pharmacological activity.
Assuntos
Antituberculosos/metabolismo , Isoniazida/análogos & derivados , Isoniazida/metabolismo , NAD/análogos & derivados , Neutrófilos/efeitos dos fármacos , Peroxidase/metabolismo , Antituberculosos/farmacologia , Humanos , Hidrazinas/química , Isoniazida/química , Isoniazida/farmacologia , NAD/química , NAD/metabolismo , Ativação de Neutrófilo/efeitos dos fármacos , Neutrófilos/citologia , Neutrófilos/enzimologia , Peroxidase/química , Cultura Primária de Células , Superóxido Dismutase/química , Superóxido Dismutase/metabolismo , Superóxidos/química , Acetato de Tetradecanoilforbol/farmacologiaRESUMO
Queen health is crucial to colony survival of social bees. Recently, queen failure has been proposed to be a major driver of managed honey bee colony losses, yet few data exist concerning effects of environmental stressors on queens. Here we demonstrate for the first time that exposure to field-realistic concentrations of neonicotinoid pesticides during development can severely affect queens of western honey bees (Apis mellifera). In pesticide-exposed queens, reproductive anatomy (ovaries) and physiology (spermathecal-stored sperm quality and quantity), rather than flight behaviour, were compromised and likely corresponded to reduced queen success (alive and producing worker offspring). This study highlights the detriments of neonicotinoids to queens of environmentally and economically important social bees, and further strengthens the need for stringent risk assessments to safeguard biodiversity and ecosystem services that are vulnerable to these substances.
Assuntos
Abelhas/efeitos dos fármacos , Isoniazida/análogos & derivados , Praguicidas/farmacologia , Animais , Abelhas/anatomia & histologia , Feminino , Isoniazida/farmacologia , MasculinoRESUMO
A simple and efficient procedure was employed for the synthesis of N'-(1,4-naphtho-quinone-2-yl) isonicotinohydrazide (NIH) by the reaction of 2-hydroxy-1,4-naphthaquinone (lawsone) and isonicotinoyl hydrazine in methanol using ultrasonic irradiation. Lawsone is the principal dye, isolated from the leaves of henna (Lawsonia inermis). Structural modification was done on the molecule aiming to get a more active derivative. The structure of the parent compound and the derivative was characterized by elemental analyses, infrared, electronic, (1)H, (13)C NMR and GC-MS spectra. The fluorescence spectral investigation of the compound was studied in DMSO and ethanol. Single crystal X-ray diffraction studies reveal that NIH crystallizes in monoclinic space group. The DNA cleavage study was monitored by gel electrophoresis method. The synthesized compound was found to have significant antioxidant activity against DPPH radical (IC50=58 µM). The in vitro cytotoxic studies of the derivative against two human cancer cell lines MCF-7 (human breast cancer) and HCT-15 (human colon carcinoma cells) using MTT assay revealed that the compound exhibited higher cytotoxic activity with a lower IC50 value indicating its efficiency in killing the cancer cells even at low concentrations. These results suggest that the structural modifications performed on lawsone could be considered a good strategy to obtain a more active drug.