Ischemic injury occurs when the flow of blood is reduced or blocked to an area of the body and can cause significant tissue damage by generation of reactive oxygen species (ROS), activation of apoptotic pathways and through induction of the inflammatory response. Restoration of blood flow and reperfusion of the blocked site, while essential, can generate a second injury that itself needs to be controlled. Together the two injuries are termed ischemia/reperfusion (I/R) injury. This type of injury is frequently encountered in medicine and is a major medical problem. Therapeutic strategies to combat I/R injury include the introduction of compounds that can scavenge ROS or can induce metabolic pathways with the effect of inhibiting apoptosis. Caffeic Acid Phenethyl Ester (CAPE), a polyphenolic compound found in propolis, has been shown to protect a variety of cells types against ROS in vitro and has also been shown to induce a variety of genes including hemeoxygenase 1 (HMOX-1) , an enzyme that has been implicated in a cytoprotective pathway. Despite showing significant cytoprotection of cells against oxidant stress in vitro, CAPE is readily hydrolyzed in plasma and is also quickly removed from circulation. This result may explain the limited cytoprotective effects of CAPE in vivo. We have synthesized a series of CAPE amide derivatives, including Caffeic Acid Phenethyl Amide (CAPA), with the aim of improving CAPE’s stability properties while maintaining the cytoprotective effects of the parent compound. We found that CAPA, in addition to 2 other amide derivatives, were able to protect human umbilical vein endothelial cells (HUVEC) against ROS to a similar degree as CAPE. In addition, we have observed significant improvement in plasma stability of CAPA over CAPE at multiple temperatures. The elimination half-life of CAPA from the systemic circulation was also seen to be significantly improved over CAPE following intravenous administration to male Sprague-Dawley rats. The longer residence time of CAPA over CAPE in circulation may potentially result in greater cytoprotection in vivo. / text

To determine the level of salt tolerance of the medicinal plant Echinacea, the physiological and biochemical characteristics of E. purpurea, E. pallida and E. angustifolia exposed to different NaCl levels (0, 50, 75, and 100 mM) were evaluated under hydroponic culture. Dry weights of shoots and roots were not affected by salinity; however E. purpurea and E. pallida exhibited higher survival rate than E. angustifolia, which also showed high salt injury index and electrolyte leakage compared to the other two species. Gas exchange (photosynthetic rate, stomatal conductance, and transpiration rate) showed a decline with increasing salt concentrations in all species with a more pronounced reduction in E. angustifolia. E. purpurea was able to retain more Na+ in the roots than the other two species showing its capacity to regulate Na+ translocation to shoots (Na+ exclusion). Moreover, the activities of two major antioxidant enzymes; superoxide dismutase (SOD) and ascorbate peroxidase (APX) were increased by salinity in E purperea, while the activities were decreased in E. angustifolia. The characteristic phytochemical profiles of caffeic acid derivatives (CADs) and alkamides/ketones were obtained for the three species, and quantitative changes were determined. Cichoric acid, the major CAD in E. purpurea, was increased with salinity up to 75 mM NaCl. A relative increase in alkamides and CADs was recorded in E. angustifolia, while in E. pallida, the level of echinacoside and major ketones (22 and 24) decreased, suggesting that the medicinal value of this species was compromised by salt stress. First evidence of salt-induced changes in alkamides and ketones in Echinacea was demonstrated in this study. Activity of phenylalanine ammonia-lyase (PAL), the major enzyme involved in the biosynthesis of CADs, was increased only in the roots of E. purpurea, further reflecting the differences in salt tolerance between species. It can be concluded that Echinacea species exhibited a limited degree of salt tolerance; however, E. purpurea showed a higher tolerance than E. pallida and E. angustifolia. This tolerance was mainly attributed to the increase in Na+ exclusion capacity, antioxidant activities and PAL activity.

Feeding sub-therapeutic levels of antibiotics to livestock has been associated withdevelopment and spread of antibiotic resistant bacteria. The present experiment was conductedto investigate the effect of antibiotic alternatives (caffeic acid, chlorogenic acid, and carbadox)on the microbial ecology of swine feces in vitro.Minimum inhibitory concentrations of caffeic and chlorogenic acids were determined forseveral pathogens using macrobroth and agar dilution techniques. Gram-negative bacteria werenot inhibited. Caffeic acid inhibited four Staphylococcus aureus strains at 200 ppm or less, andtwo Clostridium perfringens strains at 300 ppm. Chlorogenic acid inhibited all S. aureus strainsat 500 ppm, and one C. perfringens strain at 400 ppm.Effects of antibiotic alternatives on fecal microbial ecology were determined using an invitro incubation. Caffeic acid lowered total anaerobes, Bifidobacteria, Escherichia coli, andpercent E. coli (pandlt;0.01). Chlorogenic acid lowered total anaerobes, Bifidobacteria, andlactobacilli (pandlt;0.01), and increased acetate concentration (pandlt;0.0001). Carbadox lowered totalanaerobes, Bifidobacteria, E. coli, and coliforms (pandlt;0.01), and lowered acetate, propionate,butyrate, valerate, and total volatile fatty acid concentrations (pandlt;0.01). It can be concluded thataddition of caffeic acid, chlorogenic acid, or carbadox effected bacterial and chemicalcomponents of the microbial ecology of swine feces.

To determine the level of salt tolerance of the medicinal plant Echinacea, the physiological and biochemical characteristics of E. purpurea, E. pallida and E. angustifolia exposed to different NaCl levels (0, 50, 75, and 100 mM) were evaluated under hydroponic culture. Dry weights of shoots and roots were not affected by salinity; however E. purpurea and E. pallida exhibited higher survival rate than E. angustifolia, which also showed high salt injury index and electrolyte leakage compared to the other two species. Gas exchange (photosynthetic rate, stomatal conductance, and transpiration rate) showed a decline with increasing salt concentrations in all species with a more pronounced reduction in E. angustifolia. E. purpurea was able to retain more Na+ in the roots than the other two species showing its capacity to regulate Na+ translocation to shoots (Na+ exclusion). Moreover, the activities of two major antioxidant enzymes; superoxide dismutase (SOD) and ascorbate peroxidase (APX) were increased by salinity in E purperea, while the activities were decreased in E. angustifolia. The characteristic phytochemical profiles of caffeic acid derivatives (CADs) and alkamides/ketones were obtained for the three species, and quantitative changes were determined. Cichoric acid, the major CAD in E. purpurea, was increased with salinity up to 75 mM NaCl. A relative increase in alkamides and CADs was recorded in E. angustifolia, while in E. pallida, the level of echinacoside and major ketones (22 and 24) decreased, suggesting that the medicinal value of this species was compromised by salt stress. First evidence of salt-induced changes in alkamides and ketones in Echinacea was demonstrated in this study. Activity of phenylalanine ammonia-lyase (PAL), the major enzyme involved in the biosynthesis of CADs, was increased only in the roots of E. purpurea, further reflecting the differences in salt tolerance between species. It can be concluded that Echinacea species exhibited a limited degree of salt tolerance; however, E. purpurea showed a higher tolerance than E. pallida and E. angustifolia. This tolerance was mainly attributed to the increase in Na+ exclusion capacity, antioxidant activities and PAL activity.

Philosophiae Doctor - PhD / This study explores the roles of exogenously applied nitric oxide, exogenously applied caffeic acid and salt stress on the antioxidant system in cereal (exemplified by maize) and legume (using soybean as an example) plants together with their influence on membrane integrity and cell death.This study investigates changes in H2O2 content, root lipid peroxidation, root cell death and antioxidant enzymatic activity in maize roots in response to exogenously applied nitric oxide (NO) and salt stress. This part of the study is based on the partially understood interaction between NO and reactive oxygen species (ROS) such as H2O2 and the role of antioxidant enzymes in plant salt stress responses. The results show that application of salt (NaCl) results in elevated levels of H2O2 and an increase in lipid peroxidation, consequently leading to increased cell death. The study also shows that by regulating the production and detoxification of ROS through modulation of antioxidant enzymatic activities, NO plays a pivotal role in maize responses to salt stress. The study argues for NO as a regulator of redox homeostasis that prevents excessive ROS accumulation during exposure of maize to salinity stress that would otherwise be deleterious to maize. This study extends the role of exogenously applied NO to improve salt stress tolerance in cereals crops (maize) further to its role in enhancing salt stress tolerance in legumes. The effect of long-term exposure of soybean to NO and salt stress on root nodule antioxidant activity was investigated to demonstrate the role of NO in salt stress tolerance. The results show that ROS scavenging antioxidative enzymes like SOD, GPX and GR are differentially regulated in response to exogenous application of NO and salt stress. It remains to be determined if the NOinduced changes in antioxidant enzyme activity under salt stress are sufficient to efficiently reduce ROS accumulation in soybean root nodules to levels close to those of unstressed soybean root nodules. Furthermore, this study investigates the effect of long-term exposure of soybean to exogenous caffeic acid (CA) and salt stress, on the basis of the established role of CA as an antioxidant and the involvement of antioxidant enzymes in plant salt stress responses. The effect of CA on soybean nodule number, biomass (determined on the basis of nodule dry weight, root dry weight and shoot dry weight), nodule NO content, and nodule cyclic guanosine monophosphate (cGMP) content in response to salt stress was investigated. Additionally, CA-induced changes in nodule ROS content, cell viability, lipid peroxidation and antioxidant enzyme activity as well as some genes that encode antioxidant enzymes were investigated in the presence or absence of salt stress. The study shows that long-term exposure of soybean to salt stress results in reduced biomass associated with accumulation of ROS, elevated levels of lipid peroxidation and elevated levels of cell death. However, exogenously applied CA reversed the negative effects of salt stress on soybean biomass, lipid peroxidation and cell death. CA reduced the salt stress-induced accumulation of ROS by mediating changes in root nodule antioxidant enzyme activity and gene expression. These CA-responsive antioxidant enzymes were found to be superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione peroxidase (GPX), and glutathione reductase (GR), which contributed to the scavenging of ROS in soybean nodules under salt stress. The work reported in Chapter 2 has been published in a peer-reviewed journal [Keyster M, Klein A, Ludidi N (2012) Caspase-like enzymatic activity and the ascorbate-glutathione cycle participate in salt stress tolerance of maize conferred by exogenously applied nitric oxide. Plant Signaling and Behavior 7: 349-360]. My contribution to the published paper was all the work that is presented in Chapter 2,whereas the rest of the work in the paper (which is not included in Chapter 2) was contributed by Dr Marshall Keyster.

Yes / Caffeic acid phenethyl ester (CAPE) is a polyphenolic active compound present in popular apiproduct, ‘propolis’ obtained from beehives. Though it has broad therapeutic capability, the bioavailability of CAPE is limited due to poor solubility. In this study, we report novel cocrystals of CAPE engineered using coformers such as caffeine (CAF), isonicotinamide (INIC), nicotinamide (NIC) with enhanced solubility and bioavailability of CAPE. The cocrystals were prepared by microwave-assisted cocrystallization and characterized using PXRD, DSC and Raman spectroscopy. PXRD and DSC confirm the successful formation and phase purity of CAPE-CAF, CAPE-INIC and CAPE-NIC cocrystals. Raman spectra of CAPE cocrystals complement these results in confirming the formation of novel crystalline phases. CAPE-NIC cocrystal was further subjected to X-ray crystallography to understand its molecular arrangement and hydrogen bonding in the crystal structure. The CAPE-NIC cocrystal structure is found to be stabilized by a rare 1,2-benzenediol-amide heterosynthon. Cocrystallization of CAPE with NIC improved its aqueous solubility and pharmacokinetic profile thereby demonstrating 2.76 folds escalation in bioavailability. / We thank UKIERI: UK-India Education and Research Initiative (TPR26) and EPSRC (EP/J003360/1, EP/L027011/1) for providing financial support during this study.

Causada por protozoários do gênero Leishmania, a leishmaniose é uma doença infecciosa que afeta milhões de pessoas em todos os continentes. São grandes os desafios no tratamento da leishmaniose porque os fármacos atualmente disponíveis apresentam alta toxicidade, efeitos colaterais, alto custo e resistência parasitária. Sendo assim, faz-se necessário o desenvolvimento de novas drogas para o tratamento da doença de maneira mais eficiente. A investigação de novas alternativas terapêuticas, novos alvos bioquímicos, em particular a arginase em de Leishmania (L) amazonensis, é considerada um alvo interessante na busca de novos compostos leishmanicidas. A via bioquímica em que está envolvida a arginase é fundamental para a manutenção do ciclo de vida do parasito, sendo responsável pela hidrólise de L-arginina em L-ornitina em mamíferos. Muitos compostos oriundos de produtos naturais tem sido relatados como inibidores da arginase de L. (L.) amazonensis e a presença de grupos hidroxila em suas estruturas mostraram ser importantes na atividade inibidora da enzima. O objetivo desse trabaho estudar a interação bioquímica entre a enzima arginase de L. (L.) amazonensis, extratos vegetais, flavonoides e derivados do ácido cafeico. O extrato acetônico de folhas de Qualea grandiflora apresentou maior capacidade inibitória da arginase (IC50 = 1,7 ± 01 µg/mL) dentre os extratos analisados. Dentre os flavonoides, o composto mais potente foi a taxifolina (IC50 = 1,7 ± 0,2 µM). O verbascosídeo foi o derivado do ácido cafeico com maior potencial de inibição da arginase (IC50 = 0,5 ± 0,2 µM). As estruturas desses compostos podem servir como referência no desenvolvimento de novas drogas para o controle da leishmaniose, que tenham como alvo a arginase. / Caused by protozoa of the genus Leishmania, leishmaniosis is an infectious disease that affects millions of people on all continents. The currently available drugs are highly toxic, expensive, have side effects and parasitic resistance and for these reasons there is a need of developing new drugs to more effective treatment of the disease. The research of new therapeutic alternatives, new biochemical targets, in particular the arginase of Leishmania (Leishmania) amazonensis is considered an interesting target in the search for new compounds leishmanicidal. The biochemistry of arginase pathway is essential for the survival of the parasite, is also responsible for hydrolysis of L- arginine to L- ornithine in mammals. Many compounds from natural products has been reported as inhibitors of arginase L. (L.) amazonensis, and the presence of hydroxyl groups in their structures shown to be important for inhibitory activity of the enzyme. This work studies the biochemical interaction between the arginase enzyme, plant extracts, flavonoids and derivatives of caffeic acid. The acetone extract of Qualea grandiflora leaves showed higher inhibitory capacity of arginase (IC50 = 1.7 ± 01 µg/mL) from the analyzed extracts. Among the flavonoids, the most potent compound was taxifolin (IC50 = 1.7 ± 0.2 µM). The verbascosideo was the derivative of caffeic acid with greater potential for arginase inhibition (IC50 = 0.5 ± 0.2 µM) The structures of these compounds can serve as a reference in developing new drugs for the control of leishmaniosis, which target arginase.

Os neutrófilos representam a primeira linha de defesa do hospedeiro, atuando na contenção e eliminação de patógenos. Contudo, alterações na vida média, no excessivo recrutamento e ativação dos neutrófilos estão associados a danos teciduais e a doenças inflamatórias e autoimunes. A modulação das funções efetoras dos neutrófilos pode auxiliar no tratamento de tais patologias. Neste sentido, os produtos naturais constituem uma importante fonte de novas substâncias imunomoduladoras. Um estudo recente demonstrou que, a inibição do metabolismo oxidativo de neutrófilos pelo extrato etanólico bruto das folhas de Baccharis dracunculifolia (EEBBd) correlaciona-se com a proporção entre ácido cafeico (CaA) e outros compostos fenólicos contidos nesta amostra. Para dar prosseguimento à investigação do potencial imunomodulador do EEBBd e do CaA, os objetivos do presente estudo foram avaliar o efeito modulador desses produtos naturais: (i) em três funções efetoras de neutrófilos humanos - fagocitose, atividade microbicida e metabolismo oxidativo estimulado por agentes independentes de receptores (forbol-12-miristato-13-acetato; PMA) e dependentes apenas de receptores Fcgama (imunocomplexos não-opsonizados; IC) ou de receptores Fcgama associados a receptores do complemento (imunocomplexos opsonizados com complemento; IC-SHN); (ii) na atividade da mieloperoxidase (MPO); (iii) na expressão de receptores de membrana; (iv) e na captura (scavenger) de H2O2 e HOCl. O CaA foi mais efetivo do que o EEBBd em inibir a atividade da MPO e em capturar H2O2 e HOCl. A análise in silico revelou que o CaA bloqueia a entrada do sítio ativo da MPO através da interação com os resíduos Gln-91, His-95 e Arg-239; os dois últimos resíduos são essenciais para clivar o H2O2 e para estabilizar o sítio ativo, respectivamente. A eficiência dos agentes utilizados para estimular o metabolismo oxidativo, medido por quimioluminescência dependente de lucigenina e de luminol, ocorreu na seguinte ordem: PMA > IC-SHN > IC. Embora o PMA tenha sido o agente mais efetivo em estimular o metabolismo oxidativo, ambas as amostras (EEBBd e CaA) inibiram com maior intensidade esta função celular estimulada por PMA do que a mesma função estimulada por IC-SHN e IC. Além disso, ambas, EEBBd e CaA, não alteraram os níveis de expressão dos receptores TLR2, TLR4, CD16, CD32, e CD11b/CD18. Nas maiores concentrações avaliadas, EEBBd (50 ug/mL) e CaA (90 ug/mL) não foram citotóxicos para os neutrófilos. O EEBBd inibiu intensamente a capacidade fagocítica e reduziu discretamente a capacidade microbicida dos neutrófilos frente à Candida albicans. Portanto, o CaA contribui para ação inibitória do EEBBd no metabolismo oxidativo e na atividade da MPO, mas não na capacidade fagocítica e microbicida de neutrófilos. Por fim, o efeito imunomodulador do CaA e do EEBBd não é mediado por alterações na viabilidade celular ou na expressão de receptores de membrana em neutrófilos. O conjunto de resultados obtidos pode auxiliar na elucidação do mecanismo de ação destes produtos naturais sobre as funções efetoras de neutrófilos, bem como no desenvolvimento de novos fármacos para o tratamento de doenças inflamatórias mediadas pela ativação exacerbada de neutrófilos. / Neutrophils represent the first line of host defense that acts in the containment and clearance of pathogens. However, changes in life span and excessive recruitment and activation of neutrophils are associated with tissue damage and inflammatory and autoimmune diseases. Modulation of the effector functions of neutrophils can help to treat these pathologies. In this sense, natural products constitute an important source of novel immunomodulating compounds. A recent study has demonstrated that the neutrophil oxidative metabolism inhibition by the crude ethanol extract of Baccharis dracunculifolia (EEBBd) leaves correlates with the ratio of caffeic acid (CaA) to other phenolic compounds that exist in it. To continue investigating the immunomodulating potential of EEBBd and CaA, the present study aimed to examine whether these natural products modulate: (i) three effector functions of human neutrophils phagocytosis, microbial killing, and oxidative metabolism elicited by a receptor-independent stimulus (phorbol-12-myristate-13-acetate; PMA) and by receptor-dependent stimuli that bind Fcgama receptors alone (non-opsonized immune complexes; IC) or in combination with complement receptors (complement-opsonized immune complexes; IC-SHN); (ii) myeloperoxidase (MPO) activity; (iii) expression of membrane receptors; (iv) H2O2 e HOCl scavenging. CaA was more effective than EEBBd in inhibiting MPO activity and scavenging H2O2 and HOCl. The in silico analysis revealed that CaA blocks the entrance of the active site of MPO through the interaction with Gln-91, His-95, and Arg-239; the two last residues are essential to cleave H2O2 and stabilize the active site, respectively. The agents used to stimulate the oxidative metabolism, as measured by the lucigenin- and luminol-dependent chemiluminescence assays, acted in the following ranking order of efficiency: PMA > IC-SHN > IC. Although PMA was the most efficient agent at stimulating the neutrophil oxidative metabolism, both samples (EEBBd and CaA) suppressed the PMA-induced oxidative metabolism more effectively than they suppressed the same cell function elicited by IC-SHN and IC. Both EEBBd and CaA did not alter the levels of TLR2, TLR4, CD16, CD32, and CD11b/CD18 receptors expression. At the highest concentrations tested, EEBBd (50 ug/mL) and CaA (90 ug/mL) were not toxic towards neutrophils. EEBBd strongly diminished the phagocytic capacity and slightly reduced the Candida albicans killing ability of neutrophils. In conclusion, CaA contributes to the EEBBd inhibitory action on the oxidative metabolism and MPO activity but not on the phagocytic capacity and microbial killing ability of neutrophils. Furthermore, the immunomodulating effect of CaA and EEBBd is not mediated by alterations in either cell viability or expression of membrane receptors in neutrophils. Together, the results can help to unravel the mechanism of action of these natural products on the effector functions of neutrophils, and to develop new drugs to treat inflammatory diseases mediated by neutrophil overactivation.

Tyrimo tikslas: ištirti kavos rūgšties fenetilo esterio (CAFE) poveikį išemijos paveiktų žiurkės inkstų mitochondrijų funkcijoms. Uždaviniai: įvertinti tiesioginį in vitro CAFE poveikį inkstų mitochondrijų funkcijoms; CAFE poveikį išemijos (20 min) in vitro paveiktų inkstų mitochondrijų funkcijoms; kvėpavimo grandinės I komplekso aktyvumui; mitochondrijų gebėjimui kaupti Ca2+. Metodai. Wistar veislės žiurkių patinėliai buvo skirstomi į 4 grupes: kontrolinė grupė, 20 min trukmės išemijos, CAFE 22 mg/kg ir CAFE 34 mg/kg. CAFE buvo leidžiamas 1,5 h prieš sukeliant išemiją. Mitochondrijos buvo išskiriamos diferencinio centrifugavimo būdu. Mitochondrijų kvėpavimo greitis mitochondrijoms oksiduojant I ir II komplekso substratus buvo registruojamas poliarografiškai Klarko tipo elektrodu. Ca2+ sugėrimas buvo matuojamas fluorimetriškai. Mitochondrijų kvėpavimo grandinės I komplekso aktyvumas buvo tiriamas spektrofotometriškai. Rezultatai: CAFE (0,7 – 4,5 M) didina mitochondrijų kvėpavimo greitį 2-oje metabolinėje būsenoje nuo 15 % iki 34 % ir neveikė mitochondrijų kvėpavimo greičio 3-ioje metabolinėje būsenoje (VADP). Didesnės koncentracijos (5,2 - 6,5 µM) slopina mitochondrijų kvėpavimo greitį VADP (16 % ir 43 % atitinkamai). Tiriant CAFE poveikį 20 min išemijos in vitro paveiktų mitochondrijų funkcijoms, nustatyta, jog 22 mg/kg ir 34 mg/kg CAFE, intraperitonealinė injekcija 1,5 h prieš sukeliant inkstų išemiją, 20 % (p<0,05) pagerino mitochondrijų kvėpavimo greitį VADP bei... [toliau žr. visą tekstą] / The aim of investigation: to analyse the effect of caffeic acid phenethyl ester (CAPE) on kidney mitochodrial functions. Objectives: to evaluate direct in vitro effect of CAPE on kidney mitochondrial functions; the impact of CAPE on ischemia (20 min) in vitro affected kidney mitochondrial functions; on the mitochondrial respiratory chain complex I activity and on the mitochondria capability to accumulate Ca2+. Methods. Wistar rats were pretreated intraperitoneal with 22 mg/kg and 34 mg/kg of CAPE. Animals were divided into 4 groups: control group, 20 min of ischemia, CAPE 22 mg/kg group and CAPE 34 mg/kg group. Mitochondria were isolated by means of differential centrifugation. The mitochondrial respiration rate while oxidizing complex I and II dependent substrates was registered polarographically with Clark-type electrode. Ca2+ accumulation was measured fluorometrically. Activity of complex I was measured spectrophotometrically. Results: the results shown that CAPE 0,7 - 4,5 µM increases mitochondrial State 2 respiration rate by 15 - 34 % and has no effect on the State 3 respiration rate. Higher concentrations (5,2 - 6,5 µM) decreased mitochondrial State 3 respiration rate by 16% and 43%, respectively. Pretreatment with CAPE (22 mg/kg and 34 mg/kg) increased (by 20%) the ischemia suppressed mitochondrial State 3 respiration rate and respiratory control index. CAPE had no effect on succinate oxydation. Pretreatment with CAPE (22 mg/kg and 34 mg/kg) increased Ca2+... [to full text]

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O complexo multienzimático NADPH oxidase é um dos componentes de maior relevância neste contexto, pois é uma das principais fontes de ânion superóxido no organismo animal. Sendo expresso em inúmeros tecidos, incluindo leucócitos e células do tecido endotelial, o desenvolvimento de inibidores eficientes deste complexo enzimático poderá significar uma nova terapêutica para o tratamento de doenças inflamatórias crônicas. Com objetivo de explorar a relação entre a estrutura molecular, as propriedades químicas e atividades biológicas, utilizamos o éster fenetílico do ácido cafeico (CAPE) como inibidor do complexo enzimático NADPH oxidase e comparamos sua eficácia com o seu precursor ácido cafeico e os derivados, éster fenetílico do ácido cinâmico e o ácido clorogênico, correlacionando-os a respeito a sua hidrofobicidade, propriedades redox e inibição do complexo NADPH oxidase em leucócitos ativados. A hipótese seria de que um aumento da hidrofobicidade provocado pela esterificação do ácido cafeico poderia facilitar o seu acesso à membrana celular e assim alterar seu efeito como possível inibidor de NADPH Oxidase. Os resultados, em ensaios in vitro, mostraram que as alterações na hidrofobicidade não provocaram alterações significativas no potencial de oxidação e potencial antioxidantes dos compostos testados. Quando testados em leucócitos ativados (modelos ex vivo), a esterificação provocou uma melhora significativa na capacidade de inibição do complexo NADPH oxidase. Este potente efeito se propagou às EROs decorrentes de ânion superóxido e produzidas por leucócitos, como peróxido de hidrogênio e ácido hipocloroso, entretanto, sem alterar a capacidade fagocítica dos leucócitos. Os resultados deste estudo mostram que nos ensaios celulares o CAPE foi o composto mais potente em relação ao seu precursor ácido e ácido clorogênico, sendo significativamente mais efetivo na inibição da produção das EROs. Da mesma forma, CAPE foi o inibidor mais eficaz da expressão de TNF-α e IL-10 por Staphylococcus aureus células estimuladas. Em conclusão, a presença do grupo catecol e a maior hidrofobicidade, do CAPE, foram essenciais para os efeitos biológicos, confirmando nossa hipótese. Considerando-se o envolvimento da NADPH-oxidases na génese e progressão de doenças inflamatórias, CAPE deve ser considerada como uma droga anti-inflamatória promissora. / Several diseases have their genesis and / or progression related to unregulated production of oxidants intermediates. The multienzymatic complex NADPH oxidase is one of the most important components in this context because it is a major source of superoxide anion in animal organisms. It is expressed in numerous tissues, including leukocytes and endothelial tissue cells. Developing effective inhibitors of this enzyme complex may indicate a new therapy for the treatment of chronic inflammatory diseases. Several studies have described numerous anti-inflammatory properties attributed to caffeic acid phenethyl ester (CAPE), an active component found in propolis. In order to explore the relationship between the molecular structure, chemical properties and biological activities, we used CAPE to inhibit the enzyme complex NADPH oxidase and compare its efficacy with the its precursor caffeic acid and derivatives, phenethyl ester of cinnamic acid and chlorogenic acid, correlating them with regard to hydrophobicity, redox properties and inhibition of NADPH oxidase complex on activated leukocytes. The hypothesis was that an increase of hydrophobicity caused by the esterification of caffeic acid could facilitate access to cell membranes and thereby alter its effect as a possible inhibitor of NADPH oxidase. The results (in vitro), showed that the changes in hydrophobicity did not provoke significant changes in the oxidation potential and antiradical potency of the tested compounds. But when tested in activated leukocytes (ex vivo), the esterification caused a significant improvement in the ability to inhibit the NADPH oxidase complex. This potent inhibition effect resulted also in the blockage of production of hypochlorous acid, however, without altering the phagocytic ability of leukocytes. The results of this study show that in cellular assays, CAPE was the most potent compound in comparison to caffeic acid and chlorogenic acid, significantly more effective in inhibiting the production of ROS. Likewise, CAPE was the most effective inhibitor of expression of TNF-α and IL-10 in Staphylococcus aureus stimulated cells. In conclusion, the presence of the catechol moiety and the higher hydrophobicity of CAPE were essential for the biological effects. Considering the involvement of NADPH oxidases in the genesis and progression of inflammatory diseases, CAPE should be considered as a promising anti-inflammatory drug.