RESUMO
Clinical studies suggest that diets rich in ω-3 polyunsaturated fatty acids (PUFAs) provide beneficial anti-inflammatory effects, in part through their conversion to bioactive metabolites. Here we report on the endogenous production of a previously unknown class of ω-3 PUFA-derived lipid metabolites that originate from the crosstalk between endocannabinoid and cytochrome P450 (CYP) epoxygenase metabolic pathways. The ω-3 endocannabinoid epoxides are derived from docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) to form epoxyeicosatetraenoic acid-ethanolamide (EEQ-EA) and epoxydocosapentaenoic acid-ethanolamide (EDP-EA), respectively. Both EEQ-EAs and EDP-EAs are endogenously present in rat brain and peripheral organs as determined via targeted lipidomics methods. These metabolites were directly produced by direct epoxygenation of the ω-3 endocannabinoids, docosahexanoyl ethanolamide (DHEA) and eicosapentaenoyl ethanolamide (EPEA) by activated BV-2 microglial cells, and by human CYP2J2. Neuroinflammation studies revealed that the terminal epoxides 17,18-EEQ-EA and 19,20-EDP-EA dose-dependently abated proinflammatory IL-6 cytokines while increasing anti-inflammatory IL-10 cytokines, in part through cannabinoid receptor-2 activation. Furthermore the ω-3 endocannabinoid epoxides 17,18-EEQ-EA and 19,20-EDP-EA exerted antiangiogenic effects in human microvascular endothelial cells (HMVEC) and vasodilatory actions on bovine coronary arteries and reciprocally regulated platelet aggregation in washed human platelets. Taken together, the ω-3 endocannabinoid epoxides' physiological effects are mediated through both endocannabinoid and epoxyeicosanoid signaling pathways. In summary, the ω-3 endocannabinoid epoxides are found at concentrations comparable to those of other endocannabinoids and are expected to play critical roles during inflammation in vivo; thus their identification may aid in the development of therapeutics for neuroinflammatory and cerebrovascular diseases.
Assuntos
Anti-Inflamatórios/sangue , Endocanabinoides/metabolismo , Compostos de Epóxi/sangue , Etanolaminas/sangue , Ácidos Graxos Ômega-3/metabolismo , Amidoidrolases/metabolismo , Animais , Encéfalo/metabolismo , Bovinos , Citocromo P-450 CYP2J2 , Sistema Enzimático do Citocromo P-450/metabolismo , Avaliação Pré-Clínica de Medicamentos , Epóxido Hidrolases/metabolismo , Compostos de Epóxi/farmacologia , Compostos de Epóxi/uso terapêutico , Etanolaminas/farmacologia , Etanolaminas/uso terapêutico , Humanos , Metabolismo dos Lipídeos , Camundongos , Microglia/metabolismo , Neovascularização Patológica/prevenção & controle , Agregação Plaquetária/efeitos dos fármacos , Ratos , Vasodilatação/efeitos dos fármacosRESUMO
Our objective was to assess the dynamics of monoepoxides derived from polyunsaturated fatty acids (MEFAs), and their response to n-3 PUFA supplementation, in the setting of acute tissue injury and inflammation (cardiac surgery) in humans. Patients (479) undergoing cardiac surgery in three countries were randomized to perioperative fish oil (EPA + DHA; 8-10 g over 2-5 days preoperatively, then 2 g/day postoperatively) or placebo (olive oil). Plasma MEFAs derived from n-3 and n-6 PUFAs were measured 2 days postoperatively. Based on serial measures in a subset of the placebo group, levels of all MEFAs declined substantially following surgery (at postoperative day 2), with declines ranging from 37% to 63% (P < 0.05 each). Compared with placebo at postoperative day 2, levels of EPA- and DHA-derived MEFAs were 40% and 18% higher, respectively (P ≤ 0.004). The n-3 PUFA supplementation did not significantly alter the decline in n-6 PUFA-derived MEFAs. Both enrollment level and changes in plasma phospholipid EPA and DHA were associated with their respective MEFAs at postoperative day 2 (P < 0.001). Under the acute stress of cardiac surgery, n-3 PUFA supplementation significantly ameliorated the reduction in postoperative n-3 MEFAs, but not n-6 MEFAs, and the degree of increase in n-3 MEFAs related positively to the circulating level of their n-3 PUFA precursors.
Assuntos
Procedimentos Cirúrgicos Cardíacos , Compostos de Epóxi/metabolismo , Ácidos Graxos Insaturados/metabolismo , Óleos de Peixe/farmacologia , Compostos de Epóxi/sangue , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
BACKGROUND: Oxylipins mediate inflammation, vascular tension, and more. Their presence in lipoproteins could explain why lipoproteins mediate nearly identical activities. METHODS: To determine how oxylipins are distributed in the lipoproteins of hypertriglyceridemic subjects, and whether omega-3 fatty acids alter them in a manner consistent with improved cardiovascular health, we recruited 15 dyslipidemic subjects whose levels of low density lipoprotein cholesterol (LDL-C) were at goal but who remained hypertriglyceridemic (200-499 mg/dL). They were treated them with the indicated dose of 4 g/d omega-3 acid ethyl esters (P-OM3) for 8 weeks. Measured oxylipins included mid-chain alcohols (HETEs, HEPEs and HDoHEs), ketones (KETEs), epoxides (as EpETrEs, EpETEs, and EpDPEs). RESULTS: At baseline, arachidonate-oxylipins (HETEs, KETEs, and EpETrEs) were most abundant in plasma with the greatest fraction of total abundance (mean |95% CI|) being carried in high density lipoproteins (HDL); 42% |31, 57| followed by very low density lipoproteins (VLDL); 27% |20, 36|; and LDL 21% |16, 28|. EPA- and DHA-derived oxylipins constituted less than 11% of total. HDL carried alcohols and epoxides but VLDL was also rich in ketones. Treatment decreased AA-derived oxylipins across lipoprotein classes (-23% |-33, -12|, p = 0.0003), and expanded EPA-(322% |241, 422|, p<0.0001) and DHA-derived oxylipins (123% |80, 176|, p<0.0001). CONCLUSIONS: Each lipoprotein class carries a unique oxylipin complement. P-OM3 treatment alters the oxylipin content of all classes, reducing pro-inflammatory and increasing anti-inflammatory species, consistent with the improved inflammatory and vascular status associated with the treatment. TRIAL REGISTRATION: ClinicalTrials.gov NCT00959842.
Assuntos
Ácidos Graxos Ômega-3/uso terapêutico , Inibidores de Hidroximetilglutaril-CoA Redutases/uso terapêutico , Hipolipemiantes/uso terapêutico , Lipoproteínas/sangue , Oxilipinas/sangue , Ácido Araquidônico/sangue , Compostos de Epóxi/sangue , Feminino , Humanos , Hipertrigliceridemia/sangue , Hipertrigliceridemia/tratamento farmacológico , Cetonas/sangue , Lipoproteínas/metabolismo , Lipoproteínas HDL/sangue , Lipoproteínas LDL/sangue , Lipoproteínas VLDL/sangue , Masculino , Pessoa de Meia-Idade , Oxilipinas/metabolismoRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Tripterygium wilfordii HOOK F (TWHF) is a traditional Chinese medicine used in the treatment of various autoimmune diseases and inflammatory disorders including rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), and skin diseases. Triptolide (TP) is one of the main active ingredients of this traditional Chinese medicine. MC002 is a novel semi-synthetic derivate of TP which is highly water soluble, acts as a prodrug and is converted to TP in vivo. AIM OF THIS STUDY: A sensitive, rapid method for the simultaneous determination of TP and its chemo-unstable prodrug MC002 in dog blood was developed and validated using electrospray ionization (ESI) liquid chromatography-tandem mass spectrometry (LC-MS/MS). Using this method, a pharmacokinetic study of MC002 and TP following an intravenous drip infusion of 0.2mg/kg MC002 in dogs was performed. MATERIALS AND METHODS: Chemo-degradation of the prodrug in blood samples was inhibited by the addition of a small amount of sodium fluoride solution before using liquid-liquid extraction with ethyl acetate. The concentrations of MC002 and TP in dog blood were determined using the LC-MS/MS method. RESULTS: The quantitative method showed good precision and stability and is suitable for the assay of biological samples. The pharmacokinetic study showed that the elimination of MC002 was faster than that of TP, and the concentrations and AUC0-t values of TP were higher than MC002. MC002 can rapidly convert to TP in vivo. CONCLUSIONS: This validated method was successfully applied in a pharmacokinetic study of MC002 following an intravenous drip infusion in dogs. With the development of this new prodrug of TP as a promising anti-cancer drug, this method is suitable for its further analysis in clinical studies.
Assuntos
Anti-Inflamatórios não Esteroides/sangue , Antineoplásicos/sangue , Diterpenos/sangue , Glicina/análogos & derivados , Fenantrenos/sangue , Pró-Fármacos/análise , Animais , Antineoplásicos/farmacocinética , Cromatografia Líquida , Diterpenos/farmacocinética , Cães , Compostos de Epóxi/sangue , Compostos de Epóxi/farmacocinética , Feminino , Glicina/sangue , Glicina/farmacocinética , Masculino , Fenantrenos/farmacocinética , Pró-Fármacos/farmacocinética , Espectrometria de Massas em TandemRESUMO
Glycidol fatty acid esters (GEs) have been identified as contaminants in refined edible oils. Although the possible release of glycidol (G) from GEs is a concern, little is known about the conversion of GEs to G in the human body. This study addressed the toxicokinetics of glycidol linoleate (GL) and G in male Crl:CD(SD) rats and cynomolgus monkeys. Equimolar amounts of GL (341 mg/kg) or G (75 mg/kg) were administered by gavage to each animal. G was found in both species after the G and GL administration, while plasma GL concentrations were below the lower limit of quantification (5 ng/ml) in both species. In rats, the administration of GL or G produced similar concentration-time profiles for G. In monkeys, the C(max) and AUC values after GL administration were significantly lower than those after G administration. The oral bioavailability of G in monkeys (34.3%) was remarkably lower than that in rats (68.8%) at 75 mg/kg G administration. In addition, plasma G concentrations after oral administration at three lower doses of GL or G were measured in both species. In monkeys, G was detected only at the highest dose of G. In contrast, the rats exhibited similar plasma G concentration-time profiles after GL or G administration with significantly higher G levels than those in monkeys. In conclusion, these results indicate that there are remarkable species differences in the toxicokinetics of GEs and G between rodents and primates, findings that should be considered when assessing the human risk of GEs.
Assuntos
Compostos de Epóxi/farmacocinética , Compostos de Epóxi/toxicidade , Ácido Linoleico/farmacocinética , Ácido Linoleico/toxicidade , Ácidos Linoleicos/farmacocinética , Ácidos Linoleicos/toxicidade , Propanóis/farmacocinética , Propanóis/toxicidade , Administração Oral , Animais , Área Sob a Curva , Disponibilidade Biológica , Diglicerídeos/química , Relação Dose-Resposta a Droga , Avaliação Pré-Clínica de Medicamentos , Compostos de Epóxi/sangue , Ácido Linoleico/sangue , Ácidos Linoleicos/sangue , Macaca fascicularis , Masculino , Propanóis/sangue , Ratos , Ratos Sprague-Dawley , Especificidade da EspécieRESUMO
In this study, 2 metabolites of imperatorin, imperatorin hydroxylate (IMH) and imperatorin epoxide (IME), were identified for the first time in dog plasma. A sensitive, specific, and accurate high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was then developed for the simultaneous quantification of imperatorin and its 2 metabolites in dog plasma. Separation was achieved on an Agilent ZORBAX Extend-C(18) column (2.1 mm × 50 mm, 3.5 µm) at 30 °C. The mobile phase consisted of 0.02% ammonium acetate solution-methanol with a gradient program at a flow rate of 0.3 mL/min. Detection was performed using an electrospray ionization source operating in positive ion multiple reaction monitoring mode and by monitoring the ion transitions from 271 to 203 m/z for imperatorin, 309.4-224.1 m/z for IMH, 287-203 m/z for IME, and 441.3-325.2 m/z for simvastatin (the internal standard). Good linearity was shown over the concentration range of 1-500 ng/mL for imperatorin, and 0.2-500 ng/mL for IMH and IME. The validated method was successfully applied to a pharmacokinetic study of imperatorin in beagle dogs. The pharmacokinetic profiles of imperatorin and its 2 metabolites showed sex differences after the i.v. administration of imperatorin at a dose of 5 mg/kg.
Assuntos
Cromatografia Líquida , Medicamentos de Ervas Chinesas/metabolismo , Medicamentos de Ervas Chinesas/farmacocinética , Furocumarinas/sangue , Furocumarinas/farmacocinética , Espectrometria de Massas em Tandem , Acetatos/química , Administração Oral , Animais , Biotransformação , Calibragem , Cromatografia Líquida/normas , Cães , Medicamentos de Ervas Chinesas/administração & dosagem , Medicamentos de Ervas Chinesas/química , Compostos de Epóxi/sangue , Fezes/química , Feminino , Furocumarinas/administração & dosagem , Furocumarinas/química , Hidroxilação , Injeções Intravenosas , Modelos Lineares , Espectroscopia de Ressonância Magnética , Masculino , Metanol/química , Estrutura Molecular , Ratos , Ratos Sprague-Dawley , Padrões de Referência , Sensibilidade e Especificidade , Fatores Sexuais , Espectrometria de Massas por Ionização por Electrospray , Espectrometria de Massas em Tandem/normasRESUMO
SCOPE: Acrylamide (AA), classified as a genotoxic carcinogen, is generated by heating foods. We studied whether the food matrix modulates bioavailability and/or biotransformation and investigated kinetics and biological effectiveness of AA in rats. METHODS AND RESULTS: AA was given to the animals at a daily intake level of AA containing foods for up to 9 days, resulting in an exposure of 50 or 100 µg AA/kg body weight (b.w.)/day. Positive controls received the same dosages of AA in water, negative controls just water. As biomarkers urinary mercapturic acids, hemoglobin adducts, plasma levels of AA and glycidamide (GA) and DNA integrity in white blood cells and hepatocytes were measured. Altogether, no significant differences in bioavailability of AA from water and the different food matrices were observed. Only with bread crust, biomarkers indicated a slightly reduced bioavailability. Monitoring glycidamide valine adduct adducts did not provide evidence for treatment-related significantly enhanced GA-haemoglobin adduct formation in blood although glycidamide mercapturic acid excretion in urine indicated significant GA formation. CONCLUSIONS: The results suggest AA at dietary intake levels, exceeding estimated human mean intake by a factor of at least 100 to become detoxified in Sprague-Dawley rats to a major extent through glutathione coupling.
Assuntos
Acrilamida/administração & dosagem , Acrilamida/toxicidade , Carcinógenos/toxicidade , Ingestão de Alimentos , Alimentos , Água/administração & dosagem , Acetilcisteína/sangue , Acetilcisteína/toxicidade , Acetilcisteína/urina , Acrilamida/sangue , Acrilamida/urina , Animais , Disponibilidade Biológica , Biomarcadores/sangue , Biotransformação , Pão , Carcinógenos/administração & dosagem , Carcinógenos/metabolismo , Dano ao DNA , Compostos de Epóxi/sangue , Compostos de Epóxi/toxicidade , Compostos de Epóxi/urina , Hemoglobinas/metabolismo , Masculino , Ratos , Ratos Sprague-Dawley , Solanum tuberosumRESUMO
The aim of this paper is to develop and validate a rapid and sensitive LC-APCI/MS method for the determination of triptolide (TP) in plasma and to study the pharmacokinetic properties of TP in Beagle dogs. Sample preparation consisted of liquid-liquid extraction of interests. with ethyl acetate from dog plasma. The analytes and internal standard prednisolone were well separated on a Zorbax Extend-C18 analytical column. Plasma TP was detected by selected-ion monitoring (SIM) of LC-APCI/MS as its deprotonated molecular ions [M - H] - at m/z 358.9. Pharmacokinetic studies were undertaken in dogs following an iv dose of 0.05 mg x kg(-1) of TP or an ig dose of 0.05, 0.08, 0.1 mg x kg(-1), separately. The pharmacokinetic parameters were calculated by DAS software. Calibration curves were linear over the concentration range of 1 - 200 ng x mL(-1) of TP with the within- and between-batch precisions less than 10%. The within and between-batch accuracy was 95.0% to 105.0%. Recovery of LC-MS method for TP in plasma was over 75%. The T1/2beta was (2.5 +/- 0.8) h after intravenous administration of TP at the dose of 0.05 mg x kg(-1). There were no significant differences in T(max), T1/2 alpha and T1/2 beta among the three ig dosage groups. AUC and C(max) increased proportionally with doses. The absolute bioavailability of TP after ig administration of 0.05 mg x kg(-1) was (75 +/- 17)%. The LC-MS method for determination of triptolide in dog plasma was sensitive and rapid. It was showed that the elimination of triptolide was rapid. The absolute bioavailability of triptolide given orally was high.