Potentiation of antinociceptive effect of NSAIDs by a specific lipooxygenase inhibitor, acetyl 11-keto-beta boswellic acid.

The present study was aimed to assess the combined effects of cyclooxygenase and 5-lipoxygenase (COX/5-LOX) inhibitors in different animal models of nociception. Naproxen, nimesulide and rofecoxib are well-established antinociceptive agents acting via COX inhibition. AKBA (acetyl-keto-beta-boswellic acid) is a 5-LOX inhibitor. AKBA (50-200 mg/kg) produced a dose dependent and significant antinociceptive effect in different animal models of nociception. Based on the earlier reports from our laboratory, sub effective doses of all the three COX Inhibitors were selected and they were administered (naproxen, 5 mg/kg; nimesulide, 1 mg/kg; and rofecoxib, 1 mg/kg) with AKBA (100 mg/kg). This produced a more significant antinociceptive effect as compared to per se effect observed in all the three models of nociception. However, the effect of combination of nimesulide with AKBA was more pronounced as compared to naproxen and rofecoxib and their combination with AKBA. The present finding provided an evidence for the potentiation of antinociceptive effect of NSAIDs with AKBA. Such a combination may help to reduce the therapeutic doses of conventional NSAIDs and also reduce side effects (gastric, cardiac and renal) that are popularly associated with the NSAIDs.

Effect of 5-lipoxygenase inhibitor against lipopolysaccharide-induced hypothermia in mice.

Bacterial endotoxin produces sepsis associated with alterations in body temperature (fever or hypothermia). The intraperitoneal administration of bacterial endotoxin, lipopolysaccharide (LPS; 50 microg/mouse) led to a decrease in colonic temperature starting 1 hr after the injection. The hypothermic effect was accompanied by a significant increase in hypothalamic leukotriene B4 (LTB4) and prostaglandin E2 (PGE2) levels. 5-lipoxygenase inhibitor, zileuton (200 and 400 mg/kg, po) administered 30 min before LPS challenge significantly prevented hypothermia. However, non-selective cyclooxygenase inhibitor, indomethacin (10, 20 mg/kg, po) did not reverse the hypothermic response. Further, pretreatment of mice with zileuton prevented LPS-stimulated increase in hypothalamic LTB4 levels and caused a relatively small increase in PGE2 levels. Indomethacin had no effect on LTB4 levels but it reduced PGE2 levels. These results suggest a possible involvement of leukotrienes in LPS-induced hypothermia and the potential protective role of 5-lipoxygenase inhibitors in endotoxemia.

Effect of 5-lipoxygenase inhibition on events associated with inflammatory bowel disease in rats.

Leukotrienes play a part in inflammatory response. The unique role of the enzyme 5-lipoxygenase (5-LOX) in the production of leukotrienes makes it a likely therapeutic target for inflammatory conditions like asthma, rheumatoid arthritis, psoriasis, and inflammatory bowel disease (IBD). The aim of the present study was to evaluate the effect of zileuton, an orally active selective 5-LOX inhibitor against the events associated with dextran sodium sulphate-induced colitis in a rat model of IBD. The animals were administered simultaneously zileuton (100mg/kg) or sulphasalazine (100mg/kg) orally for 7 days. On day eight, rats were sacrificed, and distal colon isolated to determine myeloperoxidase activity, in vivo superoxide dismutase activity, prostaglandin E2 levels and histological examination. Both zileuton and sulphasalazine significantly prevented the development of inflammatory events associated with colitis. The effect of zileuton was more pronounced towards reducing myeloperoxidase activity and increasing PGE2 levels in distal colon. The results show that chemotactic leukotrienes are responsible for inflammatory surge in damaged colon and, zileuton, significantly improved healing by inhibition of neutrophil recruitment and indirectly through increase in prostaglandins at the site of inflammation. It is suggested that inhibitors of 5-LOX enzyme may have useful therapeutic role in the treatment of chronic intestinal inflammation.

Enzyme selectivity of new cyclooxygenase-2/5 lipoxygenase inhibitors using molecular modeling approach.

We have studied the conformational flexibility of three 5-keto-substituted 7-tert-butyl-2,3-dihydro-3,3-dimethylbenzofurans (DHDMBFs) which show dual cyclooxygenase (COX) and 5-lipoxygenase (LOX) inhibition and are potential candidates as antiinflammatory agents and analgesics. The conformations were studied by systematic search, molecular mechanics (MM) and simulated annealing molecular dynamics (SAMD) techniques. We also studied several structure based parameters and distribution of molecular electrostatic potential (MEP) around these molecules. All the three compounds were docked in the active cavity of cyclooxygenase-2 (COX-2) using graphical and energy grid search techniques. The complex geometries were optimized by MM. The results on conformational flexibility, inter-atomic distances and angles, MEP distribution and points of contacts with peptide side chains in active cavity have been used to understand the mechanistic cause of differential action of these molecules.

Effecto del clonixinato de lisina y la indometacina sobre la actividad de la lipooxigenasa y la ciclooxigenasa de colon aislado de pacientes con neoplasias / Effect of lysine clonixinate and indomethacin on the lipooxygenase and cycloogygenase activity of clon isolated from cancer patients

Las ulceraciones de la mucosa gástrica y colónica producida por los antiinflamatorios no esteroides (AINES) son producidas por la reducción de la síntesis de prostaglandinas (PGs) por inhibición de la ciclooxigenasa. Por otro lado se ha demosntrado que los productos de la 5-lipoxigenasa (5-LO) son agentes ulcerogénicos. En algunos casos el uso de AINES se acompaña de una mayor producción de productos de la 5-LO debido a un exceso de sustrato (ácido araquidónico) al estar bloqueada la ciclooxigenasa. Cuando ocurre esto último aumentan los efectos lesivos de los AINES ya que no sólo disminuyen las PGS citoprotectoras, sino que también aumentan los productos de la 5-LO. El objeto de este trabajo, fue estudiar en segmentos de colon aislado de pacientes con neoplasias el efecto del Clonixinato de lisina (CL) y de la indometacina (INDO) sobre la síntesis de PGs e hidroxiácidos (HETEs). Las concentraciones de CL (4 y 6 mug/ml) y de indo (0,035 y 0,035 y 0,35 mug/ml utilizadas, se corresponden con los valores plasmáticos alcanzados con dosis terapéuticos orales de ambas drogas. Se observó que el CL con ninguna de las dosis utilizadas modificó la producción basal de PGE(2). Por el contrario la INDO inhibió significativamente la síntese de PGE(2) con ambas dosis. Llamó la atención que el CL tanto a 4 mug/ml como 6 mug/ml produjo una profunda disminución en la producción de 5-HETE, efecto sólo observado con la concentración más alta de INDO. Este resultado indicaría una probable acción inhibitoria sobre la 5-LO, primeira enzima del camino metabólico del ácido araquidónico hacia formación de HETEs y Lts. Concluimos que el CL en dosis terapéuticas tiene un mecanismo de acción diferente al de los AINES clásicos. Los datos obtenidos en este estudo explicarían la baja incidencia de efectos gastrointestinales lesivos observados con CL.

Synthesis of hydroxy ureas as inhibitors of 5-lipoxygenase

A novel series of N-aryl-N-hydroxyurea derivatives [III] was synthesized for possible biological activity as inhibitors of 5-lipoxygenase

Effect of lipoxygenase and cyclo-oxygenase inhibition on oleic acid-induced lung oedema

Luego de una infusión de ácido oleico (AO) durante 10 minutos, se estudió la contribución de los metabolitos secundarios de la cicloxigénasa y de la lipoxigenasa en la hemodinamia y en la formación de edema en 21 pulmones aislados de conejo, mediante el registro de los cambios de la Tasa de Filtración de Líquido (TFL) y de la Presión de la Arteria Pulmonar (PAP). Cincuenta minutos antes de la infusión de AO se petrataron 7 pulmones con indometacina (inhibidor de cicloxigenasa), y 7 con diestilcarbamacina (inhibidor de lipoxigenasa). Siete no fueron retratados (grupo control). A los 12 minutos después de la infusión de AO (7.6ñ2.3 mg x min vs. 2.3ñ0.8 mg x min**-1 y 0.96ñ0.8 mg x min**-1 respectivamente) (p<0.01) se obtuvo que la TFL en el grupo de indometacina resultó mucho mayor que en el grupo control de indometacina resultó mucho mayor que en el grupo control y que en el grupo tratado con dietilcarbamacina (DEC). A los 20 minutos de la infusión, la TFL en los pulmones control resultó significativamente mayor que el valor correspondiente al grupo de DEC (4.2ñ0.5 mg x min vs. 1.6ñ1.0 mg x min**-1) (p<0.01). La Presión Media de la Arteria Pulmonar (PMAP) aumentó tanto en el grupo control como en el grupo de indometacina (de 16.0ñ2.0 Torr a 24.3ñ3.7 Torr luego de transcurrir 20 minutos de la infusión de AO y de 14.4ñ2.5 Torr a 24.6ñ3.6 Torr a los 10 minutos de la infusión de AO respectivamente); sin embargo, el valor de PMAP en el grupo de DEC no tuvo un cambio significativo al transcurrir 30 minutos luego de la infusión de AO (de 14.7ñ1.5 Torr a 16.0 ñ2.3 Torr) (p>0.05). Por lo tanto, concluimos que la inhibición selectiva de los metabolitos de la 5-lipoxigenasa (leucotrienos) pueden desempeñar un papel protector en el edema inducido por AO, mientras que la inhibición selectiva de la cicloxigenasa puede traer como consecuencia un efecto nocivo en la permeabilidad endotelial y en la hemodinamia en nuestra condición experimental