RESUMO
Tyrosinase is a copper-containing enzyme involved in the biosynthesis of melanin pigment. While the excess production of melanin causes hyperpigmentation of human skin, hypopigmentation results in medical conditions like vitiligo. Tyrosinase inhibitors could be used as efficient skin whitening agents and tyrosinase agonists could be used for enhanced melanin synthesis and skin protection from UV exposure. Among a wide range of tyrosinase-regulating compounds, natural and synthetic derivatives of furochromenones, such as 8-methoxypsoralen (8-MOP), are known to both activate and inhibit tyrosinase. We recently reported a synthetic approach to generate a variety of dihydrofuro[3,2-c]chromenones and furo[3,2-c]chromenones in a metal-free condition. In the present study, we investigated these compounds for their potential as antagonists or agonists of tyrosinase. Using fungal tyrosinase-based in vitro biochemical assay, we obtained one compound (3k) which could inhibit tyrosinase activity, and the other compound (4f) that stimulated tyrosinase activity. The kinetic studies revealed that compound 3k caused 'mixed' type tyrosinase inhibition and 4f stimulated the catalytic efficiency. Studying the mechanisms of these compounds may provide a basis for the development of new effective tyrosinase inhibitors or activators.
Assuntos
Inibidores Enzimáticos , Monofenol Mono-Oxigenase , Monofenol Mono-Oxigenase/antagonistas & inibidores , Monofenol Mono-Oxigenase/metabolismo , Inibidores Enzimáticos/farmacologia , Inibidores Enzimáticos/química , Inibidores Enzimáticos/síntese química , Cinética , Humanos , Metoxaleno/farmacologia , Metoxaleno/química , Ativadores de Enzimas/química , Ativadores de Enzimas/farmacologiaRESUMO
Divergence of gene function is a hallmark of evolution, but assessing such divergence in one species or between species requires information on functional alterations of the alleles and homologs. Here, we explore the functional divergence of two paralogs, CYP6AE19 and CYP6AE20, from Helicoverpa armigera, and two close orthologs, CYP6B8 and CYP6B7, from two related species (Helicoverpa zea and H. armigera); although there is high sequence identity within each pair of enzymes, the latter P450 of each pair has lost metabolic competence towards the plant allelochemical xanthotoxin. Multiple chimeric and single/double site mutants were created by exchanging the diverse substrate recognition sites (SRSs) and amino acids within each pair of P450s. Heterologous expression in Sf9 cells and in vitro metabolism studies showed that the exchange of SRS4 swapped the activity of CYP6AE19 and CYP6AE20, and subsequent site-directed mutagenesis demonstrated that the CYP6AE20 V318M substitution causes a gain-of-function towards xanthotoxin. Meanwhile, a single amino acid substitution (L489P) in SRS6 was found to swap activity between the CYP6B orthologs. Sequence alignments of CYP6AE paralogs and all reported insect xanthotoxin-metabolizing P450s suggest M318 and P489 are essential for the catalytic activities of CYP6AE paralogs and CYP6B orthologs, respectively, but P450s in different subfamilies may have different mechanisms towards the same substrate. Our findings demonstrate that a single amino acid substitution can suffice to alter substrate metabolism and this functional divergence resulting from natural mutations will help to further our understanding of the process of natural selection of P450 genes and their role in insect-host plant interactions.
Assuntos
Inseticidas , Mariposas , Aminoácidos , Animais , Sistema Enzimático do Citocromo P-450/metabolismo , Insetos/metabolismo , Inseticidas/metabolismo , Metoxaleno/química , Metoxaleno/metabolismo , Mariposas/metabolismoRESUMO
Acetylcholinesterase (AChE) inhibitor is the first choice for the treatment of Alzheimer's disease (AD), but it has some defects, such as dose limitation and unsatisfactory long-term treatment effect. Recent studies have shown that butyrylcholinesterase (BuChE) inhibitors or double acetyl and butyryl cholinesterase inhibitors have better curative effects on AD, and the side effects are lower than those of specific AChE inhibitors. Dual target cholinesterase inhibitors have become a new hotspot in the research of anti-AD drugs. Herein, the synthesis and bioactivities of BuChE inhibitors were reviewed.
Assuntos
Butirilcolinesterase/farmacologia , Inibidores da Colinesterase/farmacologia , Acridinas/química , Acridinas/farmacologia , Doença de Alzheimer/tratamento farmacológico , Butirilcolinesterase/efeitos adversos , Butirilcolinesterase/química , Inibidores da Colinesterase/efeitos adversos , Inibidores da Colinesterase/química , Humanos , Metoxaleno/análogos & derivados , Metoxaleno/química , Metoxaleno/farmacologia , Relação Estrutura-AtividadeRESUMO
PUVA is a treatment that combines oral methoxypsoralen (8-MOP) with ultraviolet radiation A (UVA). It is used for severe psoriasis and the early stages of T-cell lymphoma. X-rays are an effective treatment for skin cancers. Both treatments are in higher doses used to treat skin malignancies and simultaneously increase the risk of keratinocyte cancer. The main objective of this study was to test whether a few PUVA or X-ray treatments could delay the development of ultraviolet radiation (UVR)-induced skin tumors in a well-established hairless mouse model. Three groups of immunocompetent mice (total, N = 75) were included in the study. All groups were UVR-exposed during the study period. In addition, one group was treated with PUVA and another group was treated with X-rays at days 45, 52, 90 and 97. A control group was treated with UVR only. We recorded when the first, second and third skin tumors were induced in each mouse. Skin tumors developed significantly earlier in both the PUVA and X-ray groups (median, 188 days) than in the control mice (median, 215 days; p < 0.001). Therefore, a few X-ray and PUVA treatments both significantly accelerated the development of skin tumors in hairless mice, compared to UVR controls. Neither treatment showed a delay of UVR-induced skin tumors and caution should be exercised before applying these treatments to sun-damaged skin.
Assuntos
Metoxaleno/efeitos adversos , Neoplasias Cutâneas/etiologia , Raios Ultravioleta , Animais , Hospedeiro Imunocomprometido , Estimativa de Kaplan-Meier , Metoxaleno/química , Metoxaleno/farmacologia , Camundongos , Camundongos Pelados , Neoplasias Induzidas por Radiação/etiologia , Pele/patologia , Neoplasias Cutâneas/mortalidade , Neoplasias Cutâneas/patologia , Pigmentação da Pele/efeitos dos fármacos , Pigmentação da Pele/efeitos da radiação , Raios XRESUMO
8-Methoxypsoralen (8-MOP) serves as a PUVA (psoralen + UV-A) agent in the treatment of certain skin diseases. Derivatives of 8-MOP with cationic aromatic substituents at the five positions were synthesized and characterized by steady-state, femtosecond and nanosecond spectroscopy as well as cyclic voltammetry. The aromatic substituents' positive charge increases the water solubility and the affinity toward intercalation into DNA. The aromatic substituents were supposed to lower the psoralen S1 energy and thereby suppress a photo-induced electron transfer (PET) with guanine-bearing DNA. Such a suppression of this PET is expected to increase the propensity of psoralens to photo-addition to DNA. For derivatives bearing methylpyridinium residues, femtosecond spectroscopy revealed an intramolecular PET occurring on the picosecond time scale. This PET precludes the population of the triplet state. As triplet states are the precursor state for the photo-addition to DNA, their intermolecular PET renders these derivatives ineffective in terms of PUVA. For two derivatives bearing trimethylphenylammonium moieties, such an intramolecular PET does not occur and the triplet state is populated. Surprisingly, these compounds also exhibit no PUVA activity. Based on these findings, implications for further optimization of PUVA agents are discussed.
Assuntos
Furocumarinas , DNA/química , Ficusina , Furocumarinas/química , Metoxaleno/química , ÁguaRESUMO
The psoralens 8-methoxypsoralen (8-MOP), 4,5',8-trimethylpsoralen (TMP) and 5-methoxypsoralen (5-MOP) find clinical application in PUVA (psoralen + UVA) therapy. PUVA treats skin diseases like psoriasis and atopic eczema. Psoralens target the DNA of cells. Upon photo-excitation psoralens bind to the DNA base thymine. This photo-binding was studied using steady-state UV/Vis and IR spectroscopy as well as nanosecond transient UV/Vis absorption. The experiments show that the photo-addition of 8-MOP and TMP involve the psoralen triplet state and a biradical intermediate. 5-MOP forms a structurally different photo-product. Its formation could not be traced by the present spectroscopic technique.
Assuntos
DNA/química , Furocumarinas/química , Metoxaleno/química , Fotoquímica/métodos , Trioxsaleno/química , Dano ao DNA , Humanos , Cinética , Preparações Farmacêuticas , Teoria Quântica , Espectrofotometria Infravermelho , Espectrofotometria Ultravioleta , Raios UltravioletaRESUMO
An efficient one-pot synthesis of 4-(1H)-quinolones through an orthogonal engagement of diverse o-haloaryl ynones with ammonia in the presence of Cu(I), involving tandem Michael addition and ArCsp2-N coupling, is presented. The substrate scope of this convenient protocol, wherein ammonium carbonate acts as both an in situ ammonia source and a base toward diverse 2-substituted 4-(1H)-quinolones, has been mapped and its efficacy validated through concise total synthesis of bioactive natural products pseudanes (IV, VII, VIII, and XII), graveoline, graveolinine, and waltherione F.
Assuntos
Amônia/química , Cetonas/química , Metoxaleno/análogos & derivados , Quinolinas/síntese química , Quinolonas/síntese química , Metoxaleno/síntese química , Metoxaleno/química , Estrutura Molecular , Quinolinas/química , Quinolonas/químicaRESUMO
A novel series of graveolinine derivatives were synthesized and evaluated as potential anti-Alzheimer agents. Compound 5f exhibited the best inhibitory activity for acetylcholinesterase (AChE) and had surprisingly potent inhibitory activity for butyrylcholinesterase (BuChE), with IC50 values of 0.72⯵M and 0.16⯵M, respectively. The results from Lineweaver-Burk plot and molecular modeling study indicated non-competitive inhibition of AChE by compound 5f. In addition, these derivatives showed potent self-induced ß-amyloid (Aß) aggregation inhibition. Moreover, 5f didn't show obvious toxicity against PC12 and HepG2 cells at 50⯵M. Finally, in vivo studies confirmed that 5f significantly ameliorates the cognitive performances of scopolamine-treated ICR mice. Therefore, these graveolinine derivatives should be thoroughly and systematically studied for the treatment of Alzheimer's disease.
Assuntos
Doença de Alzheimer/tratamento farmacológico , Inibidores da Colinesterase/farmacologia , Metoxaleno/análogos & derivados , Acetilcolinesterase/metabolismo , Doença de Alzheimer/metabolismo , Peptídeos beta-Amiloides/antagonistas & inibidores , Animais , Comportamento Animal/efeitos dos fármacos , Butirilcolinesterase/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Inibidores da Colinesterase/síntese química , Inibidores da Colinesterase/química , Relação Dose-Resposta a Droga , Electrophorus , Células Hep G2 , Cavalos , Humanos , Masculino , Metoxaleno/síntese química , Metoxaleno/química , Metoxaleno/farmacologia , Camundongos , Estrutura Molecular , Células PC12 , Fragmentos de Peptídeos/antagonistas & inibidores , Ratos , Relação Estrutura-AtividadeRESUMO
A comprehensive cocrystal study for the insoluble natural pharmaceutical compound xanthotoxin (XT) was conducted, in which xanthotoxin-para aminobenzoic acid (XT-PABA) and xanthotoxin-oxalic acid (XT-OA) cocrystals were obtained. The xanthotoxin cocrystals were characterized by powder X-ray diffraction, thermal analysis, and FT-IR spectra, and the crystal structures were determined by single-crystal X-ray diffraction. Crystal structures and thermal analysis showed that XT-OA was more stable than XT-PABA. Energy framework calculation indicated that H-bond and π···π interactions generated in XT-OA were stronger than that in XT-PABA and xanthotoxin. The powder dissolution experiments of xanthotoxin and its cocrystals suggested the XT-OA cocrystal might be applied as an alternative formulation of API, on account of its enhanced solubility and stability in the hydrochloric acid buffer solution (pH 1.2). The cocrystallization engineering can prolong the enhanced apparent solubility via modulating the stability.
Assuntos
Metoxaleno/química , Solubilidade/efeitos dos fármacos , Ácido 4-Aminobenzoico/química , Cristalização/métodos , Cristalografia por Raios X/métodos , Estabilidade de Medicamentos , Ligação de Hidrogênio , Ácido Oxálico/química , Difração de Pó/métodos , Pós/química , Difração de Raios X/métodosRESUMO
Two DNA-cross-linking reagents, bis-chloroethylnitrosourea and 8-methoxypsoralen, are used to covalently cross-link interstrand base pairs in DNA bonds that, in part, define colloidal crystals engineered with DNA. The irreversible linkages formed increase the chemical and thermal stability of the crystals and do not significantly affect their long-range order, as evidenced by small-angle X-ray scattering data. The post-modified crystals are stable in environments that the pre-modified structures are not, including solvents that encompass a broad range of polarities from ethanol to hexanes, and in aqueous media at pH 0 and 14. Interestingly, the cross-linked DNA bonds within these crystals still retain their flexibility, which is reflected by a solvent-dependent reversible change in lattice parameter. Since these organic cross-linking reagents, in comparison with inorganic approaches (use of silver ions or SiO2), have marginal effects on the composition and properties of the crystals, they provide an attractive alternative for stabilizing colloidal crystals engineered with DNA and make them potentially useful in a broader range of media.
Assuntos
Reagentes de Ligações Cruzadas/química , DNA/química , Etilnitrosoureia/química , Ouro/química , Metoxaleno/química , Coloides/química , Cristalização , Etilnitrosoureia/análogos & derivados , Difração de Nêutrons , Tamanho da Partícula , Espalhamento a Baixo Ângulo , Propriedades de SuperfícieRESUMO
Linear furocoumarins, also known as psoralens, are clinically useful photo-activated pharmaceuticals employed to address hyperproliferative skin diseases. Seven diverse cytotoxic pharmacophores have been synthetically attached to 8-methoxypsoralen via a 5-amino functionality. The resulting unique set of compounds was evaluated for dark and light toxicity against PAM212 keratinocytes in culture.
Assuntos
Sobrevivência Celular/efeitos dos fármacos , Escuridão , Luz , Metoxaleno/farmacologia , Fármacos Fotossensibilizantes/farmacologia , Células Cultivadas , Humanos , Queratinócitos/efeitos dos fármacos , Queratinócitos/efeitos da radiação , Metoxaleno/química , Fármacos Fotossensibilizantes/química , Dermatopatias/patologiaRESUMO
Anethum sowa L. has been used as a spice herb in the Asian and European culinary systems to add flavour and taste. The studied plant has diverse folkloric medicinal value. Present study was designed to isolate phytochemicals from the hexane, chloroform and ethyl acetate extracts of the roots by various chromatographic techniques. Based on spectral analysis (IR, LC-MS, NMR) the isolated compounds were identified as physcione (1), ß-sitosterol (2), stigmasterol (3), 2-oxo-3-propyl-2H-chromene-7-carboxylic acid (4), bergapten (5), 3-ethyl-7-hydroxy-2H-chromen-2-one (6) and graveolone (7). The mentioned compounds have been isolated for the first time from the roots part of the plant. Based on extensive literature review, physcione and bergapten were inferred to exhibit crucial bioactivities including inhibitory efficacy against various forms of cancer. Accordingly, in the present research approach molecular docking investigations of the isolated phytochemicals have been robustly executed with different oncogenes that have been reported to be actively involved in various forms of carcinoma. In silico investigations encompassing molecular docking analysis and drug-likeness profiling was executed to estimate the potential therapeutic tendencies of the phytochemicals targeted towards effective cancer therapy. Current investigation offers meaningful know-how pertaining to potential anticancer activities of the phytochemicals extracted from the roots of Anethum sowa L. and might open up new revenues towards effective drug development against cancer.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Apiaceae/química , Simulação de Acoplamento Molecular , Extratos Vegetais/farmacologia , Raízes de Plantas/química , 5-Metoxipsoraleno/química , 5-Metoxipsoraleno/isolamento & purificação , 5-Metoxipsoraleno/farmacologia , Antineoplásicos Fitogênicos/química , Antineoplásicos Fitogênicos/isolamento & purificação , Benzopiranos/química , Benzopiranos/isolamento & purificação , Benzopiranos/farmacologia , Ácidos Carboxílicos/química , Ácidos Carboxílicos/isolamento & purificação , Ácidos Carboxílicos/farmacologia , Proliferação de Células/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Metoxaleno/análogos & derivados , Metoxaleno/química , Metoxaleno/isolamento & purificação , Metoxaleno/farmacologia , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Sitosteroides/química , Sitosteroides/isolamento & purificação , Sitosteroides/farmacologia , Estigmasterol/química , Estigmasterol/isolamento & purificação , Estigmasterol/farmacologiaRESUMO
The objective is to evaluate methoxsalen as an in vitro phenotyping tool in comparison to ABT as a nonspecific inactivator of P450 mediated metabolism. The reversible inhibition of methoxsalen and ABT against the P450, FMO, AO, MAO-A and -B, enzymes were evaluated using standard marker probe reactions. The time-dependent inhibition of P450 enzymes was evaluated in human liver microsomes. CES1 activities were determined by monitoring the depletion of known substrate, the clopidogrel. The metabolism of P450 substrates in the presence and absence of methoxsalen or ABT was evaluated in human liver microsomes. Methoxsalen is a direct inhibitor and inhibited the activities (>90%) of all enzymes at a concentration of 300 µM except for CYP2C9. Methoxsalen is also a potent time-dependent inhibitor of all P450 enzymes except for CYP2C19 (moderate) at a concentration of 300 µM. Methoxsalen inhibited the metabolism of P450 substrates in the pre-incubation mode. ABT is a potent TDI of several P450 except for CYP2C19 (47%) and CYP2C9 (27%). The results indicate that methoxsalen is a potent pan P450 inhibitor than ABT and can be a better tool in distinguishing P450 mediated metabolism form non-P450 metabolism in human liver microsomes.
Assuntos
Sistema Enzimático do Citocromo P-450/química , Metoxaleno/química , Microssomos Hepáticos/metabolismo , Triazóis/química , Clopidogrel/metabolismo , Inibidores das Enzimas do Citocromo P-450 , Humanos , Fenótipo , Isoformas de Proteínas/antagonistas & inibidoresRESUMO
This study aimed to verify the applicability of a proposed photosafety screening system based on a reactive oxygen species (ROS) assay and a cassette-dosing pharmacokinetic (PK) study to chemicals with wide structural diversity. The orally taken chemicals, erythromycin, gatifloxacin, 8-methoxypsoralen (MOP), pirfenidone (PFD), trifluoperazine (TFP), and voriconazole (VRZ), were selected as test compounds. The ROS assay was conducted to evaluate their photoreactivity, and all test compounds excluding erythromycin generated significant ROS under simulated sunlight exposure. According to the ROS data, TFP had potent photoreactivity, and the photoreactivity of 4 other compounds was judged to be moderate. Regarding the oral cassette-dosing PK test in rats, the skin deposition of MOP, PFD, and VRZ was relatively high, and gatifloxacin and TFP exhibited moderate skin deposition properties. Based on the ROS and PK data of test compounds, PFD and TFP were judged to be potent phototoxic compounds, and MOP and VRZ were deduced to have phototoxic risk. The predicted phototoxic risk of test compounds by proposed screening was mostly in agreement with observed in vivo phototoxicity in the rat skin. The proposed screening system could provide reliable photosafety information on orally administered compounds with wide structural diversity.
Assuntos
Dermatite Fototóxica/diagnóstico , Pele/efeitos dos fármacos , Testes de Toxicidade Aguda/métodos , Administração Oral , Animais , Dermatite Fototóxica/etiologia , Dermatite Fototóxica/patologia , Dermatite Fototóxica/prevenção & controle , Estudos de Viabilidade , Masculino , Metoxaleno/administração & dosagem , Metoxaleno/química , Metoxaleno/toxicidade , Piridonas/administração & dosagem , Piridonas/química , Piridonas/toxicidade , Ratos , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/metabolismo , Medição de Risco/métodos , Pele/metabolismo , Pele/efeitos da radiação , Relação Estrutura-Atividade , Distribuição Tecidual , Trifluoperazina/administração & dosagem , Trifluoperazina/química , Trifluoperazina/toxicidade , Raios Ultravioleta/efeitos adversos , Voriconazol/administração & dosagem , Voriconazol/química , Voriconazol/toxicidadeRESUMO
The navel orangeworm (Amyelois transitella) and the fungus Aspergillus flavus constitute a facultative mutualism and pest complex in tree nut and fruit orchards in California. The possibility exists that the broad detoxification capabilities of A. flavus benefit its insect associate by metabolizing toxicants, including hostplant phytochemicals and pesticides. We examined this hypothesis by conducting laboratory bioassays to assess growth rates and survivorship of pyrethroid-resistant (R347) and susceptible (CPQ) larval strains on potato dextrose agar diet containing almond meal with and without two furanocoumarins, xanthotoxin and bergapten, found in several hostplants, and with and without two insecticides, bifenthrin and spinetoram, used in almond and pistachio orchards. Additionally, fungi were incubated in liquid diets containing the test chemicals, and extracts of these diets were added to almond potato dextrose agar (PDA) diets and fed to larvae to evaluate the ability of the fungus to metabolize these chemicals. Larvae consuming furanocoumarin-containing diet experienced higher mortality than individuals on unamended diets, but adding A. flavus resulted in up to 61.7% greater survival. Aspergillus flavus in the diet increased development rate > two-fold when furanocoumarins were present, demonstrating fungal enhancement of diet quality. Adding extracts of liquid diets containing xanthotoxin and fungus decreased mortality compared to xanthotoxin alone. On diets containing bifenthrin and spinetoram, however, mortality increased. These results support the hypothesis that A. flavus enhances navel orangeworm performance and contributes to detoxification of xenobiotics. Among practical implications of our findings, this mutualistic association should be considered in designing chemical management strategies for these pests.
Assuntos
Aspergillus flavus/metabolismo , Mariposas/efeitos dos fármacos , Micotoxinas/toxicidade , 5-Metoxipsoraleno/química , 5-Metoxipsoraleno/toxicidade , Animais , Tolerância a Medicamentos , Larva/efeitos dos fármacos , Larva/crescimento & desenvolvimento , Macrolídeos/química , Macrolídeos/toxicidade , Metoxaleno/química , Metoxaleno/toxicidade , Mariposas/crescimento & desenvolvimento , Micotoxinas/químicaRESUMO
Eight alkaloids (1â»8) were isolated from Ruta graveolens, and their herbicide activities were evaluated through in vitro, semivivo, and in vivo assays. The most relevant results were observed for Compounds 5 and 6â»8 at 150 µM, which decreased dry biomass by 20% and 23%, respectively. These are significant results since they presented similar values with the positive control, commercial herbicide 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU). Based on the performed assays, Compound 5 (graveoline) is classified as an electron-transport inhibitor during the light phase of photosynthesis, as well as a plant-growth regulator. On the other hand, Compounds 6â»8 inhibited electron and energy transfers, and are also plant-growth inhibitors. These phytotoxic behaviors based on acridone and quinolone alkaloids may serve as a valuable tool in the further development of a new class of herbicides since natural products represent an interesting alternative to replace commercial herbicides, potentially due their low toxicity.
Assuntos
Alcaloides/isolamento & purificação , Metoxaleno/análogos & derivados , Fotossíntese/efeitos dos fármacos , Ruta/química , Alcaloides/química , Alcaloides/farmacologia , Biomassa , Diurona/farmacologia , Transporte de Elétrons , Herbicidas/farmacologia , Metoxaleno/química , Metoxaleno/isolamento & purificação , Metoxaleno/farmacologiaRESUMO
Oral therapy with 8-methoxypsoralen (8-MOP) may cause major side effects, whereas the topical treatment might not be much effective due to the low penetration induced by typical formulations. Therefore, the objectives of this work are the development and characterization of a nanoemulsion (NE) containing 8-MOP together with an ex vivo permeation study, monitored by a validated HPLC-Fluo method, to determine the amount of drug retained in viable skin (epidermis (E) and dermis (D)) and in stratum corneum (SC). The optimized conditions for NE formulation were achieved by full factorial designs (25 and 32): 60â¯s and 60% of ultrasound time and potency, respectively; 10â¯mL of final volume; 2% v/v of oil phase (clove essential oil); and 10% m/v of Poloxamer 407. The NE showed mean droplet diameter of 24.98⯱â¯0.49â¯nm, polydispersity index (PDI) of 0.091⯱â¯0.23, pH values of 6.54⯱â¯0.06, refractive index of 1.3525⯱â¯0.0001 and apparent viscosity of 51.15⯱â¯3.66â¯mPa at 20⯰C. Droplets with nanospherical diameters were also observed by transmission electron microscopy (TEM). Ex vivo permeation study showed that 8.5% of the applied 8-MOP dose permeated through the biological membranes, with flux (J) of 1.35⯵gâ¯cm-2â¯h-1. The drug retention in Eâ¯+â¯D and in SC was 10.15⯱â¯1.36 and 1.95⯱â¯0.71⯵gâ¯cm-2, respectively. Retention in viable skin induced by the NE was almost two-fold higher than a compounded cream (5.04⯱â¯0.30⯵gâ¯cm-2). These results suggested that the developed NE is a promising alternative for 8-MOP topical therapy when compared to commercial formulations.
Assuntos
Metoxaleno/administração & dosagem , Nanopartículas/administração & dosagem , Fármacos Fotossensibilizantes/administração & dosagem , Pele/metabolismo , Administração Cutânea , Animais , Óleo de Cravo/administração & dosagem , Óleo de Cravo/química , Óleo de Cravo/farmacocinética , Composição de Medicamentos , Sistemas de Liberação de Medicamentos , Estabilidade de Medicamentos , Emulsões , Metoxaleno/química , Metoxaleno/farmacocinética , Nanopartículas/química , Permeabilidade , Fármacos Fotossensibilizantes/química , Fármacos Fotossensibilizantes/farmacocinética , Poloxâmero/administração & dosagem , Poloxâmero/química , Poloxâmero/farmacocinética , Absorção Cutânea , Solubilidade , SuínosRESUMO
Coumarins are abundant in Umbelliferae and Rutaceae plants possessing varied pharmacological activities. The objectives of this study are to develop and validate the method for determination of six coumarins in rat plasma by liquid chromatography coupled with tandem mass spectrometry (LC-MS) and identify the metabolites of bergapten by ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS), respectively. Data-dependent acquisition mode (DDA) was applied to trigger enhanced product ion (EPI) scans by analyzing multiple reaction monitoring (MRM) signals. An efficient data processing method "key product ions (KPIs)" was used for rapid detection and identification of metabolites as an assistant tool. The time to reach the maximum plasma concentration ( Tmax) for the six compounds ranged from 1 to 6 h. A total of 24 metabolites of bergapten were detected in vitro and in vivo. The results could provide a basis for absorption and metabolism of coumarins.
Assuntos
Medicamentos de Ervas Chinesas/química , Metoxaleno/análogos & derivados , 5-Metoxipsoraleno , Animais , Cromatografia Líquida de Alta Pressão , Cumarínicos/sangue , Cumarínicos/química , Cumarínicos/farmacocinética , Medicamentos de Ervas Chinesas/farmacocinética , Metoxaleno/sangue , Metoxaleno/química , Metoxaleno/farmacocinética , Ratos , Ratos Sprague-Dawley , Espectrometria de Massas em TandemRESUMO
Bergapten (5-methoxypsoralen), a coumarin-derivate compound isolated from Ficus hirta roots, was evaluated for its anti-inflammatory and proresolution activities in a tail-cutting-induced zebrafish larvae model. Bergapten was evaluated using a caudal fin-wounded transgenic zebrafish line "Tg(corola: eGFP)" to visualize the effects of the recruitment and clearance of neutrophils and macrophages at the injury site. We found that bergapten significantly suppressed the recruitment of neutrophils and macrophages toward the injury site, as well as promoted the clearance of neutrophils and macrophages from the wound site. We also investigated the reactive oxygen species (ROS) and nitric oxide (NO) level of bergapten in a tail-cutting-induced inflammation zebrafish model. The Results revealed that bergapten effectively inhibited the tail-cutting-induced production of ROS and NO in zebrafish larvae. This study reported for the first time the potential anti-inflammatory and proresolution activities of bergapten in an in vivo zebrafish model, suggesting that bergapten may be a potential candidate for inflammation therapy.
Assuntos
Anti-Inflamatórios/farmacologia , Inflamação/tratamento farmacológico , Macrófagos/efeitos dos fármacos , Metoxaleno/análogos & derivados , Neutrófilos/efeitos dos fármacos , Cicatrização/efeitos dos fármacos , 5-Metoxipsoraleno , Animais , Anti-Inflamatórios/química , Modelos Animais de Doenças , Feminino , Ficus/química , Inflamação/imunologia , Macrófagos/imunologia , Masculino , Metoxaleno/química , Metoxaleno/farmacologia , Neutrófilos/imunologia , Óxido Nítrico/antagonistas & inibidores , Óxido Nítrico/imunologia , Raízes de Plantas/química , Espécies Reativas de Oxigênio/antagonistas & inibidores , Espécies Reativas de Oxigênio/imunologia , Peixe-Zebra/imunologiaRESUMO
Methoxsalen (MTS) is a natural bioactive compound found in a variety of plants that has many known biofunctions; however, its effects on osteoporosis and related mechanisms are not clear. This study examined whether MTS exhibited preventive effects against postmenopausal osteoporosis. Female C3H/HeN mice were divided into four groups: Sham, ovariectomy (OVX), OVX with MTS (0.02% in diet), and OVX with estradiol (0.03 µg/day, s.c). After 6 weeks, MTS supplementation significantly increased femur bone mineral density and bone surface along with bone surface/total volume. MTS significantly elevated the levels of serum formation markers (estradiol, osteocalcin and bone-alkaline phosphatase) such as estradiol in OVX mice. Tartrate resistant acid phosphatase staining revealed that MTS suppressed osteoclast numbers and formation in femur tissues compared with the OVX group. Supplementation of MTS slightly up-regulated osteoblastogenesis-related genes (Runx-2, osterix, osteocalcin, and Alp) expression, whereas it significantly down-regulated inflammatory genes (Nfκb and Il6) expression in femur tissue compared with the OVX group. These results indicate that MTS supplementation effectively prevented OVX-induced osteoporosis via enhancement of bone formation and suppression of inflammatory response in OVX mice. Our study provides valid scientific information regarding the development and application of MTS as a food ingredient, a food supplement or an alternative agent for preventing postmenopausal osteoporosis.