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1.
Environ Pollut ; 337: 122508, 2023 Nov 15.
Artículo en Inglés | MEDLINE | ID: mdl-37673322

RESUMEN

The toxicity of acrylamide (AA) has continuously attracted wide concerns as its extensive presence from both environmental and dietary sources. However, its hepatic metabolic transformation and metabolic fate still remain unclear. This study aims to unravel the metabolic profile and glutathione (GSH) mediated metabolic fate of AA in liver of rats under the dose-dependent exposure. We found that exposure to AA dose-dependently alters the binding of AA and GSH and the generation of mercapturic acid adducts, while liver as a target tissue bears the metabolic transformation of AA via regulating GSH synthesis and consumption pathways, in which glutamine synthase (GSS), cytochrome P450 2E1 (CYP2E1), and glutathione S-transferase P1 (GSTP1) play a key role. In response to high- and low-dose exposures to AA, there were significant differences in liver of rats, including the changes in GSH and cysteine (CYS) activities and the conversion ratio of AA to glycidamide (GA), and liver can affect the transformation of AA by regulating the GSH-mediated metabolic pathway. Low-dose exposure to AA activates GSH synthesis pathway in liver and upregulates GSS activity and CYS content with no change in γ-glutamyl transpeptidase 1 (GGT1) activity. High-dose exposure to AA activates the detoxification pathway of GSH and increases GSH consumption by upregulating GSTP1 activity. In addition, molecular docking results showed that most of the metabolic molecules transformed by AA and GA other than themselves can closely bind to GSTP1, GSS, GGT1, N-acetyltransferase 8, and dimethyl sulfide dehydrogenase 1. The binding of AA-GSH and GA-GSH to GSTP1 and CYP2E1 enzymes determine the tendentiousness between toxicity and detoxification of AA, which exerts a prospective avenue for targeting protective role of hepatic enzymes against in vivo toxicity of AA.


Asunto(s)
Acrilamida , Citocromo P-450 CYP2E1 , Ratas , Animales , Acrilamida/toxicidad , Acrilamida/metabolismo , Simulación del Acoplamiento Molecular , Estudios Prospectivos , Acetilcisteína/metabolismo , Glutatión Transferasa/metabolismo , Hígado/metabolismo , Metaboloma , Glutatión/metabolismo , Compuestos Epoxi/metabolismo
2.
Biomed Khim ; 68(3): 177-189, 2022 Jun.
Artículo en Ruso | MEDLINE | ID: mdl-35717582

RESUMEN

Cyclooxygenase and lipoxygenase derived lipid metabolites of polyunsaturated fatty acids (PUFAs), as well as their role in the inflammation, have been studied quite thoroughly. However, cytochrome P450 derived lipid mediators, as well as their participation in the regulation of the inflammation, need deeper understanding. In recent years, it has become known that PUFAs are oxidized by cytochrome P450 epoxygenases to epoxy fatty acids, which act as the extremely powerful lipid mediators involved in resolving inflammation. Recent studies have shown that the anti-inflammatory mechanisms of ω-3 PUFAs are also mediated by their conversion to the endocannabinoid epoxides. Thus, it is clear that a number of therapeutically relevant functions of PUFAs are due to their conversion to PUFA epoxides. However, with the participation of cytochrome P450 epoxygenases, not only PUFA epoxides, but also other metabolites are formed. They are further are converted by epoxide hydrolases into pro-inflammatory dihydroxy fatty acids and anti-inflammatory dihydroxyeicosatrienoic acids. The study of the role of PUFA epoxides in the regulation of the inflammation and pharmacological modeling of the activity of epoxide hydrolases are the promising strategies for the treatment of the inflammatory diseases. This review systematizes the current literature data of the fatty acid epoxides, in particular, the endocannabinoid epoxides. Their role in the regulation of inflammation is discussed.


Asunto(s)
Compuestos Epoxi , Ácidos Grasos Omega-3 , Antiinflamatorios , Citocromo P-450 CYP2J2 , Sistema Enzimático del Citocromo P-450/metabolismo , Endocannabinoides/metabolismo , Epóxido Hidrolasas/metabolismo , Compuestos Epoxi/metabolismo , Compuestos Epoxi/farmacología , Ácidos Grasos , Ácidos Grasos Omega-3/metabolismo , Ácidos Grasos Omega-3/farmacología , Humanos , Inflamación/tratamiento farmacológico
3.
Mol Med Rep ; 21(6): 2303-2310, 2020 06.
Artículo en Inglés | MEDLINE | ID: mdl-32323812

RESUMEN

Celastrol and triptolide, chemical compounds isolated from Tripterygium wilfordii hook (also known as thunder god vine), are effective against rheumatoid arthritis (RA). Celastrol targets numerous signaling pathways involving NF­κB, endoplasmic reticulum Ca2+­ATPase, myeloid differentiation factor 2, toll­like receptor 4, pro­inflammatory chemokines, DNA damage, cell cycle arrest and apoptosis. Triptolide, inhibits NF­κB, the receptor activator of NF­κB (RANK)/RANK ligand/osteoprotegerin signaling pathway, cyclooxygenase­2, matrix metalloproteases and cytokines. The present review examined the chemistry and bioavailability of celastrol and triptolide, and their molecular targets in treating RA. Clinical studies have demonstrated that T. wilfordii has several promising bioactivities, but its multi­target toxicity has restricted its application. Thus, dosage control and structural modification of T. wilfordii are required to reduce the toxicity. In this review, future directions for research into these promising natural products are discussed.


Asunto(s)
Diterpenos/química , Fenantrenos/química , Tripterygium/química , Triterpenos/química , Artritis Reumatoide/tratamiento farmacológico , Artritis Reumatoide/patología , Quimiocinas/antagonistas & inhibidores , Quimiocinas/metabolismo , Ciclooxigenasa 2/química , Ciclooxigenasa 2/metabolismo , Diterpenos/metabolismo , Diterpenos/farmacología , Diterpenos/uso terapéutico , Compuestos Epoxi/química , Compuestos Epoxi/metabolismo , Compuestos Epoxi/farmacología , Compuestos Epoxi/uso terapéutico , Humanos , FN-kappa B/antagonistas & inhibidores , FN-kappa B/metabolismo , Triterpenos Pentacíclicos , Fenantrenos/metabolismo , Fenantrenos/farmacología , Fenantrenos/uso terapéutico , Transducción de Señal/efectos de los fármacos , Receptor Toll-Like 4/antagonistas & inhibidores , Receptor Toll-Like 4/metabolismo , Tripterygium/metabolismo , Triterpenos/metabolismo , Triterpenos/farmacología , Triterpenos/uso terapéutico
4.
Chembiochem ; 21(14): 1976-1980, 2020 07 16.
Artículo en Inglés | MEDLINE | ID: mdl-32181956

RESUMEN

Human drug-metabolizing cytochrome P450 monooxygenases (CYPs) have enormous substrate promiscuity; this makes them promising tools for the expansion of natural product diversity. Here, we used CYP3A4 for the targeted diversification of a plant biosynthetic route leading to monoterpenoid indole alkaloids. In silico, in vitro and in planta studies proved that CYP3A4 was able to convert the indole alkaloid vinorine into vomilenine, the former being one of the central intermediates in the ajmaline pathway in the medicinal plant Rauvolfia serpentina (L.) Benth. ex Kurz. However, to a much larger extent, the investigated conversion yielded vinorine (19R,20R)-epoxide, a new metabolite with an epoxide functional group that is rare for indole alkaloids. The described work represents a successful example of combinatorial biosynthesis towards an increase in biodiversity of natural metabolites. Moreover, characterisation of the products of the in vitro and in planta transformation of potential pharmaceuticals with human CYPs might be indicative of the route of their conversion in the human organism.


Asunto(s)
Citocromo P-450 CYP3A/metabolismo , Rauwolfia/química , Alcaloides de Triptamina Secologanina/metabolismo , Compuestos Epoxi/química , Compuestos Epoxi/metabolismo , Humanos , Alcaloides Indólicos/química , Alcaloides Indólicos/metabolismo , Modelos Moleculares , Conformación Molecular , Rauwolfia/metabolismo , Alcaloides de Triptamina Secologanina/química , Estereoisomerismo , Especificidad por Sustrato
5.
Int J Mol Sci ; 21(5)2020 Mar 09.
Artículo en Inglés | MEDLINE | ID: mdl-32182938

RESUMEN

Hepatocellular carcinoma (HCC) is a leading cause of cancer death. The multikinase inhibitor sorafenib is widely used for systemic therapy in advanced HCC. Sorafenib might affect epoxyeicosanoids, as it is also a potent inhibitor of the soluble epoxide hydrolase (sEH), which catalyzes the conversion of epoxides derived from long-chain polyunsaturated fatty acids (PUFAs), such as arachidonic acid (AA) and omega-3 docosahexaenoic acid (DHA), into their corresponding diols. Experimental studies with AA-derived epoxyeicosatrienoic acids (EETs) showed that they can promote tumor growth and metastasis, while DHA-derived 19,20-epoxydocosapentaenoic acid (19,20-EDP) was shown to have anti-tumor activity in mice. In this pilot study, we assessed the effect of sorafenib treatment on the presence of lipid mediators, such as EETs, in blood of the patients with HCC using the lipidomics technology. We found a significant increase in 11,12-EET and 14,15-EET levels in HCC patients treated with sorafenib. Furthermore, while not significant in this small sample set, the data presented indicate that sorafenib can also increase the level of omega-3 DHA-derived 19,20-EDP. While the effect on EETs might hamper the anti-tumor effect of sorafenib, we hypothesize that supplementation of DHA in sorafenib-treated HCC patients could increase the level of 19,20-EDP and thereby enhance its anti-tumor effect.


Asunto(s)
Carcinoma Hepatocelular/tratamiento farmacológico , Carcinoma Hepatocelular/metabolismo , Ácidos Grasos Omega-3/metabolismo , Ácidos Grasos Omega-6/metabolismo , Neoplasias Hepáticas/tratamiento farmacológico , Neoplasias Hepáticas/metabolismo , Sorafenib/uso terapéutico , Anciano , Antineoplásicos/uso terapéutico , Ácido Araquidónico/metabolismo , Ácidos Docosahexaenoicos/metabolismo , Eicosanoides/metabolismo , Epóxido Hidrolasas/metabolismo , Compuestos Epoxi/metabolismo , Femenino , Humanos , Masculino , Persona de Mediana Edad , Proyectos Piloto
6.
Nat Commun ; 11(1): 971, 2020 02 20.
Artículo en Inglés | MEDLINE | ID: mdl-32080175

RESUMEN

Triptolide is a trace natural product of Tripterygium wilfordii. It has antitumor activities, particularly against pancreatic cancer cells. Identification of genes and elucidation of the biosynthetic pathway leading to triptolide are the prerequisite for heterologous bioproduction. Here, we report a reference-grade genome of T. wilfordii with a contig N50 of 4.36 Mb. We show that copy numbers of triptolide biosynthetic pathway genes are impacted by a recent whole-genome triplication event. We further integrate genomic, transcriptomic, and metabolomic data to map a gene-to-metabolite network. This leads to the identification of a cytochrome P450 (CYP728B70) that can catalyze oxidation of a methyl to the acid moiety of dehydroabietic acid in triptolide biosynthesis. We think the genomic resource and the candidate genes reported here set the foundation to fully reveal triptolide biosynthetic pathway and consequently the heterologous bioproduction.


Asunto(s)
Sistema Enzimático del Citocromo P-450/genética , Sistema Enzimático del Citocromo P-450/metabolismo , Diterpenos/metabolismo , Fenantrenos/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Tripterygium/genética , Tripterygium/metabolismo , Abietanos/metabolismo , Antineoplásicos Fitogénicos/biosíntesis , Vías Biosintéticas/genética , Medicamentos Herbarios Chinos/metabolismo , Compuestos Epoxi/metabolismo , Perfilación de la Expresión Génica , Genoma de Planta , Humanos , Ingeniería Metabólica , Metaboloma , Oxidación-Reducción , Filogenia , Plantas Medicinales/genética , Plantas Medicinales/metabolismo
7.
Biomolecules ; 10(1)2020 01 09.
Artículo en Inglés | MEDLINE | ID: mdl-31936432

RESUMEN

Chirality is a common phenomenon, and it is meaningful to explore interactions between stereoselective bio-macromolecules and chiral small molecules with preclinical and clinical significance. Protopanaxadiol-type ginsenosides are main effective ingredients in ginseng and are prone to biotransformation into a pair of ocotillol C20-24 epoxide epimers, namely, (20S,24S)-epoxy-dammarane-3,12,25-triol (24S-PDQ) and (20S,24R)-epoxy dammarane-3,12,25-triol (24R-PDQ) that display stereoselective fate in vivo. However, possible molecular mechanisms involved are still unclear. The present study aimed to investigate stereoselective ADME (absorption, distribution, metabolism and excretion) characteristics of PDQ epimers based on molecular docking analysis of their interaction with some vital proteins responsible for drug disposal. Homology modeling was performed to obtain 3D-structure of the human isoenzyme UGT1A8, while calculation of docking score and binding free energy and ligand-protein interaction pattern analysis were achieved by using the Schrödinger package. Stereoselective interaction was found for both UGT1A8 and CYP3A4, demonstrating that 24S-PDQ was more susceptible to glucuronidation, whereas 24R-PDQ was more prone to oxidation catalyzed by CYP3A4. However, both epimers displayed similarly strong interaction with P-gp, a protein with energy-dependent drug-pump function, suggesting an effect of the dammarane skeleton but not C-24 stereo-configuration. These findings provide an insight into stereo-selectivity of ginsenosides, as well as a support the rational development of ginseng products.


Asunto(s)
Citocromo P-450 CYP3A/metabolismo , Compuestos Epoxi/metabolismo , Glucuronosiltransferasa/metabolismo , Sapogeninas/metabolismo , Miembro 1 de la Subfamilia B de Casetes de Unión a ATP/química , Miembro 1 de la Subfamilia B de Casetes de Unión a ATP/metabolismo , Dominio Catalítico , Citocromo P-450 CYP3A/química , Compuestos Epoxi/química , Glucurónidos/química , Glucurónidos/metabolismo , Glucuronosiltransferasa/química , Humanos , Simulación del Acoplamiento Molecular , Oxidación-Reducción , Panax/química , Panax/metabolismo , Sapogeninas/química , Estereoisomerismo , Triterpenos/química , Triterpenos/metabolismo , Damaranos
8.
Prostaglandins Other Lipid Mediat ; 147: 106385, 2020 04.
Artículo en Inglés | MEDLINE | ID: mdl-31698143

RESUMEN

Cytochrome P450 (CYP) metabolism of arachidonic acid (ARA) produces epoxy fatty acids (EpFAs) such as epoxyeicosatrienoic acids (EETs) that are known to exert protective effects in inflammatory disorders. Endogenous EpFAs are further metabolized into corresponding diols by the soluble epoxide hydrolase (sEH). Through inhibition of sEH, many studies have demonstrated the cardioprotective and renoprotective effects of EpFAs; however, the role of sEH inhibition in modulating the pathogenesis of neuroinflammatory disorders is less well described. In this review, we discuss the current knowledge surrounding the effects of sEH inhibition and EpFA action in neuroinflammatory disorders such as Parkinson's Disease (PD), stroke, depression, epilepsy, and Alzheimer's Disease (AD), as well as the potential mechanisms that underlie the therapeutic effects of sEH inhibition.


Asunto(s)
Fármacos del Sistema Nervioso Central/farmacología , Enfermedades del Sistema Nervioso Central/tratamiento farmacológico , Sistema Enzimático del Citocromo P-450/metabolismo , Epóxido Hidrolasas/antagonistas & inhibidores , Compuestos Epoxi/metabolismo , Ácidos Grasos/metabolismo , Animales , Enfermedades del Sistema Nervioso Central/metabolismo , Modelos Animales de Enfermedad , Evaluación Preclínica de Medicamentos , Epóxido Hidrolasas/metabolismo , Humanos
9.
Chin J Nat Med ; 17(8): 575-584, 2019 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-31472894

RESUMEN

3-Hydroxy-3-methylglutaryl-CoA synthase (HMGS) is the first committed enzyme in the MVA pathway and involved in the biosynthesis of terpenes in Tripterygium wilfordii. The full-length cDNA and a 515 bp RNAi target fragment of TwHMGS were ligated into the pH7WG2D and pK7GWIWG2D vectors to respectively overexpress and silence, TwHMGS was overexpressed and silenced in T. wilfordii suspension cells using biolistic-gun mediated transformation, which resulted in 2-fold increase and a drop to 70% in the expression level compared to cells with empty vector controls. During TwHMGS overexpression, the expression of TwHMGR, TwDXR and TwTPS7v2 was significantly upregulated to the control. In the RNAi group, the expression of TwHMGR, TwDXS, TwDXR and TwMCT visibly displayed downregulation to the control. The cells with TwHMGS overexpressed produced twice higher than the control value. These results proved that differential expression of TwHMGS determined the production of triptolide in T. wilfordii and laterally caused different trends of relative gene expression in the terpene biosynthetic pathway. Finally, the substrate acetyl-CoA was docked into the active site of TwHMGS, suggesting the key residues including His247, Lys256 and Arg296 undergo electrostatic or H-bond interactions with acetyl-CoA.


Asunto(s)
Diterpenos/metabolismo , Regulación de la Expresión Génica de las Plantas , Hidroximetilglutaril-CoA Sintasa/genética , Hidroximetilglutaril-CoA Sintasa/metabolismo , Fenantrenos/metabolismo , Proteínas de Plantas/metabolismo , Tripterygium/metabolismo , Acetilcoenzima A/metabolismo , Secuencia de Aminoácidos , Vías Biosintéticas , Dominio Catalítico , Compuestos Epoxi/metabolismo , Hidroximetilglutaril-CoA Sintasa/química , Modelos Moleculares , Triterpenos Pentacíclicos , Proteínas de Plantas/química , Proteínas de Plantas/genética , Interferencia de ARN , Terpenos/metabolismo , Tripterygium/enzimología , Tripterygium/genética , Triterpenos/metabolismo
10.
Planta ; 250(5): 1613-1620, 2019 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-31388830

RESUMEN

MAIN CONCLUSION: A novel GA13-oxidase ofTripterygium wilfordii, TwGA13ox, is a 2-oxoglutarate-dependent dioxygenase. It specifically catalyzes the conversion of GA9to GA20, but not GA4to GA1. Gibberellins (GAs) play essential roles in plant growth and development. Previous characterization of GA20- and GA3-oxidases yielded a large number of genetic elements that can interconvert different GAs. However, enzymes that catalyze the 13-hydroxylation step are rarely identified. Here, we report that the GA13-oxidase of Tripterygium wilfordii, TwGA13ox, is a 2-oxoglutarate-dependent dioxygenase instead of reported cytochrome P450 oxygenases, among 376 differential proteins in comparative proteomics. Phylogenetic analysis showed that the enzyme resides in its own independent branch in the DOXC class. Unexpectedly, it specifically catalyzes the conversion of GA9 to GA20, but not GA4 to GA1. Contrary to the previous research, TwGA13ox transcriptional expression was upregulated ~ 146 times by exogenous application of methyl jasmonate (MeJA). RNAi targeting of TwGA13ox in T. wilfordii led to an 89.9% decrease of triptolide, a diterpenoid epoxide with extensive anti-inflammatory and anti-tumor properties. In subsequent MeJA supplementation experiments, triptolide production increased 13.4-times. TwGA13ox displayed root-specific expression. Our results provide a new GA13-oxidase from plants and elucidate the metabolic associations within the diterpenoid biosynthetic pathway (GAs, triptolide) at the genetic level.


Asunto(s)
Acetatos/farmacología , Ciclopentanos/farmacología , Dioxigenasas/metabolismo , Regulación Enzimológica de la Expresión Génica , Giberelinas/metabolismo , Oxidorreductasas/metabolismo , Oxilipinas/farmacología , Tripterygium/enzimología , Vías Biosintéticas , Dioxigenasas/genética , Diterpenos/metabolismo , Compuestos Epoxi/metabolismo , Regulación de la Expresión Génica de las Plantas , Ácidos Cetoglutáricos/metabolismo , Oxidorreductasas/genética , Fenantrenos/metabolismo , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Tripterygium/genética
11.
Biol Chem ; 400(7): 939-950, 2019 06 26.
Artículo en Inglés | MEDLINE | ID: mdl-30844759

RESUMEN

Since ancient times, people have been attracted by dyes and they were a symbol of power. Some of the oldest dyes are indigo and its derivative Tyrian purple, which were extracted from plants and snails, respectively. These 'indigoid dyes' were and still are used for coloration of textiles and as a food additive. Traditional Chinese medicine also knows indigoid dyes as pharmacologically active compounds and several studies support their effects. Further, they are interesting for future technologies like organic electronics. In these cases, especially the indigo derivatives are of interest but unfortunately hardly accessible by chemical synthesis. In recent decades, more and more enzymes have been discovered that are able to produce these indigoid dyes and therefore have gained attention from the scientific community. In this study, group E monooxygenases (styrene monooxygenase and indole monooxygenase) were used for the selective oxygenation of indole (derivatives). It was possible for the first time to show that the product of the enzymatic reaction is an epoxide. Further, we synthesized and extracted indigoid dyes and could show that there is only minor by-product formation (e.g. indirubin or isoindigo). Thus, group E monooxygenase can be an alternative biocatalyst for the biosynthesis of indigoid dyes.


Asunto(s)
Biocatálisis , Colorantes/química , Carmin de Índigo , Oxigenasas de Función Mixta/metabolismo , Biotransformación , Compuestos Epoxi/metabolismo , Indoles/metabolismo
12.
Nat Prod Res ; 33(19): 2837-2844, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-30513208

RESUMEN

New aurone epoxide, 2,10-oxy-10-methoxysulfuretin (14), and new auronolignan (15), named cotinignan A, were isolated by silica gel column and semipreparative HPLC chromatography from the methylene chloride/methanol extract of Cotinus coggygria Scop. heartwood. In addition, thirteen known secondary metabolites namely sulfuretin, 2,3-trans-fustin, fisetin, butin, butein, taxifolin, eriodictyol, 3',5,5',7-tetrahydroxyflavanone, 3',4',7-trihydroxyflavone, 3-O-methyl-2,3-trans-fustin, 3-O-galloyl-2,3-trans-fustin, ß-resorcylic acid and 3-O-ß-sitosterol glucoside were isolated as well. Their structures were elucidated by 1D and 2D NMR, HR-ESI-MS, IR and UV. Ten out of eleven isolated flavonoids possess 7, 3' and 4' hydroxy groups. These structural features could be considered as chemotaxonomic characteristic of flavonoids from C. coggygria. Cotinignan A (15) represents new subclass of secondary metabolites - auronolignans.


Asunto(s)
Anacardiaceae/química , Benzofuranos/química , Compuestos Epoxi/química , Madera/química , Anacardiaceae/metabolismo , Benzofuranos/metabolismo , Chalconas/química , Chalconas/metabolismo , Cromatografía Líquida de Alta Presión , Compuestos Epoxi/metabolismo , Flavanonas/química , Flavanonas/metabolismo , Flavonoides/análisis , Flavonoides/química , Flavonoides/metabolismo , Flavonoles , Hidroxibenzoatos/química , Hidroxibenzoatos/metabolismo , Estructura Molecular , Extractos Vegetales/química , Metabolismo Secundario , Madera/metabolismo
13.
Chem Biol Interact ; 281: 32-36, 2018 Feb 01.
Artículo en Inglés | MEDLINE | ID: mdl-29273563

RESUMEN

Acorus calamus has a rich history in natural medicine, and offers many health benefits. The plant has anti-inflammatory, antimicrobial, diuretic, antiurolithiatic and other properties. Moreover, various parts, especially the rhizome and roots, are sources of a range of bioactive phenolic compounds with beneficial effects on the cardiovascular system. This review article summarizes the current knowledge of the chemical composition of different parts of A. calamus and their roles in the prevention and treatment of cardiovascular diseases. However, as no human studies have been performed, the review only includes in vitro and animal studies. The paper also briefly reviews the toxicity of A. calamus and its products for human health, especially regarding the cardiovascular system.


Asunto(s)
Enfermedades Cardiovasculares/prevención & control , Aceites Volátiles/uso terapéutico , Acorus/química , Acorus/metabolismo , Derivados de Alilbenceno , Animales , Anisoles/química , Anisoles/metabolismo , Anisoles/uso terapéutico , Peso Corporal/efectos de los fármacos , Enfermedades Cardiovasculares/tratamiento farmacológico , Compuestos Epoxi/química , Compuestos Epoxi/metabolismo , Compuestos Epoxi/toxicidad , Humanos , Hipolipemiantes/química , Hipolipemiantes/uso terapéutico , Aceites Volátiles/química , Aceites Volátiles/metabolismo , Fenoles/química , Fenoles/uso terapéutico
14.
Plant J ; 93(1): 50-65, 2018 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-29086455

RESUMEN

Tripterygium wilfordii, which has long been used as a medicinal plant, exhibits impressive and effective anti-inflammatory, immunosuppressive and anti-tumor activities. The main active ingredients are diterpenoids and triterpenoids, such as triptolide and celastrol, respectively. A major challenge to harnessing these natural products is that they are found in very low amounts in planta. Access has been further limited by the lack of knowledge regarding their underlying biosynthetic pathways, particularly for the abeo-abietane tri-epoxide lactone triptolide. Here suspension cell cultures of T. wilfordii were found to produce triptolide in an inducible fashion, with feeding studies indicating that miltiradiene is the relevant abietane olefin precursor. Subsequently, transcriptome data were used to identify eight putative (di)terpene synthases that were then characterized for their potential involvement in triptolide biosynthesis. This included not only biochemical studies which revealed the expected presence of class II diterpene cyclases that produce the intermediate copalyl diphosphate (CPP), along with the more surprising finding of an atypical class I (di)terpene synthase that acts on CPP to produce the abietane olefin miltiradiene, but also their subcellular localization and, critically, genetic analysis. In particular, RNA interference targeting either both of the CPP synthases, TwTPS7v2 and TwTPS9v2, or the subsequently acting miltiradiene synthase, TwTPS27v2, led to decreased production of triptolide. Importantly, these results then both confirm that miltiradiene is the relevant precursor and the relevance of the identified diterpene synthases, enabling future studies of the biosynthesis of this important bioactive natural product.


Asunto(s)
Transferasas Alquil y Aril/metabolismo , Diterpenos/metabolismo , Fenantrenos/metabolismo , Tripterygium/enzimología , Transferasas Alquil y Aril/genética , Vías Biosintéticas , Compuestos Epoxi/metabolismo , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Medicinales , Interferencia de ARN , Tripterygium/genética
15.
Theranostics ; 7(8): 2250-2260, 2017.
Artículo en Inglés | MEDLINE | ID: mdl-28740548

RESUMEN

The weakened tumour colonization of attenuated Salmonella has severely hampered its clinical development. In this study, we investigated whether an anti-inflammation and antiangiogenesis compound triptolide could improve the efficacy of VNP20009, a highly attenuated Salmonella strain, against mice melanoma. By comparing the effects of conventional VNP20009 monotherapy and a combination therapy that uses both triptolide and VNP20009, we found that triptolide significantly improved the tumour colonization of VNP20009 by reducing the number of infiltrated neutrophils in the melanoma, which led to a larger necrotic area in the melanoma. Moreover, the combination therapy suppressed tumour angiogenesis by reducing the expression of VEGF in a synergistic manner, retarding the growth of the melanoma. Our study revealed that triptolide could significantly enhance the antitumour effect of VNP20009 by modulating tumour angiogenesis and the host immune response, providing a new understanding of the strategy to improve Salmonella-mediated tumour therapy.


Asunto(s)
Diterpenos/metabolismo , Diterpenos/farmacología , Inmunosupresores/metabolismo , Inmunosupresores/farmacología , Melanoma/terapia , Fenantrenos/metabolismo , Fenantrenos/farmacología , Salmonella/efectos de los fármacos , Salmonella/crecimiento & desarrollo , Animales , Terapia Biológica/métodos , Terapia Combinada/métodos , Modelos Animales de Enfermedad , Compuestos Epoxi/metabolismo , Compuestos Epoxi/farmacología , Melanoma/microbiología , Melanoma/patología , Ratones Endogámicos C57BL , Necrosis , Neovascularización Patológica/tratamiento farmacológico , Neutrófilos/inmunología , Resultado del Tratamiento , Factor A de Crecimiento Endotelial Vascular/antagonistas & inhibidores
16.
Phytother Res ; 31(7): 1090-1096, 2017 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-28509400

RESUMEN

Triptolide (TP) is an active ingredient isolated from Tripterygium wilfordii Hook. f. (TWHF), which is a traditional herbal medicine widely used for the treatment of rheumatoid arthritis and autoimmune disease in the clinic. However, its adverse reactions of hepatotoxicity and nephrotoxicity have been frequently reported which limited its clinical application. The aim of this study was to investigate the mechanism of glycyrrhetinic acid (GA) effecting on the elimination of TP in HK-2 cells and the role of the efflux transporters of P-gp and multidrug resistance-associated proteins (MRPs) in this process. An ultra performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC-ESI-MS) analytical method was established to determine the intracellular concentration of TP. In order to study the role of efflux transporters of P-gp and MRPs in GA impacting on the accumulation of TP, the inhibitors of efflux transporters (P-gp: verapamil; MRPs: MK571) were used in this study. The results showed that GA could enhance the elimination of TP and reduce the TP accumulation in HK-2 cells. Verapamil and MK571 could increase the intracellular concentration of TP; in addition, GA co-incubation with verapamil significantly increased the TP cellular concentration compared with the control group. In conclusion, GA could reduce the accumulation of TP in HK-2 cells, which was related to P-gp. This is probably one of the mechanisms that TP combined with GA to detoxify its toxicity. Copyright © 2017 John Wiley & Sons, Ltd.


Asunto(s)
Miembro 1 de la Subfamilia B de Casetes de Unión a ATP/metabolismo , Diterpenos/metabolismo , Ácido Glicirretínico/farmacología , Proteínas Asociadas a Resistencia a Múltiples Medicamentos/metabolismo , Fenantrenos/metabolismo , Línea Celular , Cromatografía Líquida de Alta Presión , Compuestos Epoxi/metabolismo , Humanos , Espectrometría de Masas , Proteína 2 Asociada a Resistencia a Múltiples Medicamentos , Plantas Medicinales/química , Propionatos/farmacología , Quinolinas/farmacología , Tripterygium/química , Verapamilo/farmacología
17.
Plant J ; 89(3): 429-441, 2017 02.
Artículo en Inglés | MEDLINE | ID: mdl-27801964

RESUMEN

Tripterygium wilfordii (Celastraceae) is a medicinal plant with anti-inflammatory and immunosuppressive properties. Identification of a vast array of unusual sesquiterpenoids, diterpenoids and triterpenoids in T. wilfordii has spurred investigations of their pharmacological properties. The tri-epoxide lactone triptolide was the first of many diterpenoids identified, attracting interest due to the spectrum of bioactivities. To probe the genetic underpinning of diterpenoid diversity, an expansion of the class II diterpene synthase (diTPS) family was recently identified in a leaf transcriptome. Following detection of triptolide and simple diterpene scaffolds in the root, we sequenced and mined the root transcriptome. This allowed identification of the root-specific complement of TPSs and an expansion in the class I diTPS family. Functional characterization of the class II diTPSs established their activities in the formation of four C-20 diphosphate intermediates, precursors of both generalized and specialized metabolism and a novel scaffold for Celastraceae. Functional pairs of the class I and II enzymes resulted in formation of three scaffolds, accounting for some of the terpenoid diversity found in T. wilfordii. The absence of activity-forming abietane-type diterpenes encouraged further testing of TPSs outside the canonical class I diTPS family. TwTPS27, close relative of mono-TPSs, was found to couple with TwTPS9, converting normal-copalyl diphosphate to miltiradiene. The phylogenetic distance to established diTPSs indicates neo-functionalization of TwTPS27 into a diTPS, a function not previously observed in the TPS-b subfamily. This example of evolutionary convergence expands the functionality of TPSs in the TPS-b family and may contribute miltiradiene to the diterpenoids of T. wilfordii.


Asunto(s)
Transferasas Alquil y Aril/genética , Liasas Intramoleculares/genética , Proteínas de Plantas/genética , Tripterygium/genética , Abietanos/química , Abietanos/metabolismo , Transferasas Alquil y Aril/clasificación , Transferasas Alquil y Aril/metabolismo , Secuencia de Aminoácidos , Diterpenos/química , Diterpenos/metabolismo , Compuestos Epoxi/química , Compuestos Epoxi/metabolismo , Perfilación de la Expresión Génica/métodos , Liasas Intramoleculares/metabolismo , Estructura Molecular , Monoterpenos/química , Monoterpenos/metabolismo , Familia de Multigenes , Fenantrenos/química , Fenantrenos/metabolismo , Filogenia , Proteínas de Plantas/metabolismo , Raíces de Plantas/enzimología , Raíces de Plantas/genética , Homología de Secuencia de Aminoácido , Tripterygium/enzimología
18.
Proc Natl Acad Sci U S A ; 113(43): 12232-12237, 2016 10 25.
Artículo en Inglés | MEDLINE | ID: mdl-27791009

RESUMEN

Macrophages are central in coordinating immune responses, tissue repair, and regeneration, with different subtypes being associated with inflammation-initiating and proresolving actions. We recently identified a family of macrophage-derived proresolving and tissue regenerative molecules coined maresin conjugates in tissue regeneration (MCTR). Herein, using lipid mediator profiling we identified MCTR in human serum, lymph nodes, and plasma and investigated MCTR biosynthetic pathways in human macrophages. With human recombinant enzymes, primary cells, and enantiomerically pure compounds we found that the synthetic maresin epoxide intermediate 13S,14S-eMaR (13S,14S-epoxy- 4Z,7Z,9E,11E,16Z,19Z-docosahexaenoic acid) was converted to MCTR1 (13R-glutathionyl, 14S-hydroxy-4Z,7Z,9E,11E,13R,14S,16Z,19Z-docosahexaenoic acid) by LTC4S and GSTM4. Incubation of human macrophages with LTC4S inhibitors blocked LTC4 and increased resolvins and lipoxins. The conversion of MCTR1 to MCTR2 (13R-cysteinylglycinyl, 14S-hydroxy-4Z,7Z,9E,11E,13R,14S,16Z,19Z-docosahexaenoic acid) was catalyzed by γ-glutamyl transferase (GGT) in human macrophages. Biosynthesis of MCTR3 was mediated by dipeptidases that cleaved the cysteinyl-glycinyl bond of MCTR2 to give 13R-cysteinyl, 14S-hydroxy-4Z,7Z,9E,11E,13R,14S,16Z,19Z-docosahexaenoic acid. Of note, both GSTM4 and GGT enzymes displayed higher affinity to 13S,14S-eMaR and MCTR1 compared with their classic substrates in the cysteinyl leukotriene metabolome. Together these results establish the MCTR biosynthetic pathway and provide mechanisms in tissue repair and regeneration.


Asunto(s)
Ácidos Docosahexaenoicos/metabolismo , Inflamación/metabolismo , Lípidos/genética , Regeneración/genética , Vías Biosintéticas/genética , Ácidos Docosahexaenoicos/genética , Compuestos Epoxi/química , Compuestos Epoxi/metabolismo , Humanos , Inflamación/genética , Metabolismo de los Lípidos/genética , Lípidos/sangre , Ganglios Linfáticos/crecimiento & desarrollo , Ganglios Linfáticos/metabolismo , Macrófagos/metabolismo , Estructura Molecular , Estereoisomerismo , Cicatrización de Heridas/genética
19.
Chembiochem ; 17(18): 1693-7, 2016 09 15.
Artículo en Inglés | MEDLINE | ID: mdl-27383542

RESUMEN

Engineered enzyme variants of potato epoxide hydrolase (StEH1) display varying degrees of enrichment of (2R)-3-phenylpropane-1,2-diol from racemic benzyloxirane. Curiously, the observed increase in the enantiomeric excess of the (R)-diol is not only a consequence of changes in enantioselectivity for the preferred epoxide enantiomer, but also to changes in the regioselectivity of the epoxide ring opening of (S)-benzyloxirane. In order to probe the structural origin of these differences in substrate selectivity and catalytic regiopreference, we solved the crystal structures for the evolved StEH1 variants. We used these structures as a starting point for molecular docking studies of the epoxide enantiomers into the respective active sites. Interestingly, despite the simplicity of our docking analysis, the apparent preferred binding modes appear to rationalize the experimentally determined regioselectivities. The analysis also identifies an active site residue (F33) as a potentially important interaction partner, a role that could explain the high conservation of this residue during evolution. Overall, our experimental, structural, and computational studies provide snapshots into the evolution of enantioconvergence in StEH1-catalyzed epoxide hydrolysis.


Asunto(s)
Biocatálisis , Evolución Molecular Dirigida , Epóxido Hidrolasas/metabolismo , Compuestos Epoxi/metabolismo , Solanum tuberosum/enzimología , Compuestos Epoxi/química , Hidrólisis , Simulación del Acoplamiento Molecular , Estructura Molecular
20.
Nat Prod Commun ; 11(5): 673-6, 2016 May.
Artículo en Inglés | MEDLINE | ID: mdl-27319148

RESUMEN

Theobroxide has been isolated from culture filtrates of Lasiodiplodia theobromae as a potato tuber-inducing compound. In this study, the metabolism of theobroxide was investigated using cowpea as an experimental model and [2H3-7]theobroxide as a substrate for analyzing a metabolite, which revealed that theobroxide applied exogenously to the roots was converted into 3-O-ß-D-glucopyranosyltheobroxide.


Asunto(s)
Ciclohexanos/metabolismo , Compuestos Epoxi/metabolismo , Fabaceae/metabolismo , Monosacáridos/metabolismo , Ascomicetos/química , Ciclohexanos/aislamiento & purificación , Compuestos Epoxi/aislamiento & purificación , Monosacáridos/aislamiento & purificación
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