Patent Abstract:
the present invention refers to thymoquinone , a main constituent of the volatile oil of nigella sativa , and its protective effect against sepsis syndrome morbidity , mortality and associated organ dysfunctions . in particular , the present invention refers to thymoquinone for use in the prevention and / or treatment of sepsis syndrome . the present invention further refers to a pharmaceutical composition and a kit .

Detailed Description:
fig1 schematically shows the main pathologic mechanisms in the early and later phases of mods . elam - 1 indicates endothelial - leukocyte adhesion molecule 1 ; paf , platelet - activating factor ; tnf - α , tumor necrosis factor - α ; vcam - 1 , vascular cell adhesion molecule 1 ( wang and ma , 2008 ). fig3 shows kaplan - meier survival analyses following lps challenge ( fig3 a ) or e . coli challenge ( fig3 b ). fig4 shows the results of biochemical blood analyses following e . coli administration . blood urea nitrogen ( marker of kidney dysfunction ) ( fig4 a ); serum creatinine ( marker of kidney dysfunction ) ( fig4 b ); alanine transferase ( marker of liver dysfunction ) ( fig4 c ); plasma albumin ( marker of oncotic pressure ) ( fig4 d ); total creatinine kinase ( marker of tissue injury ) ( fig4 e ); total lactate dehydrogenase ( marker of tissue injury ) ( fig4 f ). fig5 shows the results of immunological serum analyses following e . coli administration . tnf - α ( fig5 a ); il - 1 α ( fig5 b ); il - 2 ( fig5 c ); il - 10 ( fig5 d ). two groups of animals ( ten mice each ) weighing ˜ 25 g were treated intraperitoneally with 1 or 2 mg / kg ( single dose ) of thymoquinone prepared in 10 % dmso ; a control group ( treated with only 10 % dmso ) was also simultaneously run . four hours after thymoquinone treatment , all groups were challenged with 2 mg / kg endotoxin ( lipopolysaccharide , lps ) prepared in normal saline . mortality of groups was followed for 4 days , and survival ( primary endpoint ) was calculated using kaplan - meier analysis . as shown in fig3 a , thymochinone provided a protective effect against mortality in endotoxin induced sepsis with about 80 % ( 1 mg / kg ) and 90 % ( 2 mg / kg ) protection efficacy , respectively ( p & lt ; 0 . 01 ). two groups of animals ( twelve mice each ) weighing ˜ 25 g were treated intraperitoneally with 0 . 75 or 1 mg / kg / day ( for three days ) of thymoquinone prepared in 10 % dmso ; a control group ( treated with only 10 % dmso ) was also simultaneously run . four hours after thymoquinone treatment , all groups were challenged with 2 μl / g e . coli ( atcc - 25992 , 1 . 5 × 10 cfu / ml ) prepared in normal saline . mortality in treated groups was followed for 2 days , and survival ( primary endpoint ) was calculated using kaplan - meier analysis . as shown in fig3 b , thymochinone provided a protective effect against mortality in sepsis induced by gram - negative bacteria with about 90 % ( 0 . 75 or 1 mg / kg ) protection efficacy ( p & lt ; 0 . 01 ). these data demonstrate that thymoquinone has a very potent protective effect against sepsis mortality that reaches up to 80 - 90 %. this activity is apparently not related to any antimicrobial activity of thymoquinone as evident by a similar protective effect seen following lps administration . in another set of experiments and after 6 h of e . coli administration , animals were lightly anesthetized with ether , and blood samples were collected , centrifuged and the resultant plasma were stored at − 80 ° c . and used within 48 h . the levels of tnf - α , il - 1α , il - 2 and il - 10 were determined using an elisa technique . important biochemical parameters including blood serum creatinine , blood urea nitrogen , alanine transferase ( alt ), albumin , and total lactate dehydrogenase ( ldh ) and creatinine kinase ( ck ) were also assessed in treated animals using commercially available kits according to the manufacturer &# 39 ; s directions . the data show that induced sepsis also causes injuries and disruption in the biochemical functions of many organs such as kidney and liver that was manifested as an elevation in serum levels of serum creatinine and blood urea nitrogen , alt , and total ldh and ck . fig4 shows the organ protective effects of thymoquinone in sepsis caused by e . coli bacteria . important reductions are seen in kidney dysfunction biomarkers ( measured by blood urea nitrogen and serum creatinine levels ), liver dysfunction biomarkers ( measured by plasma albumin and alaninine transferase ) with improvement in albumin level , and in total creatinine kinase and lactate dehydrogenease ( markers for tissue injury ). fig5 shows the immunomodulatory effects of thymoquinone in sepsis induced by e . coli bacteria . significant reductions are seen in the levels of mediators and biomarkers of inflammation including tnf - α , il - 1α , il - 2 and il - 10 . allometric scaling relates dosing information to total body weight across species using a power equation . given an allometric relationship in animals , one can use this relationship to predict a starting dose in man that may yield desirable exposures : dose man = dose animal *( wt man / wt animal ) 0 . 75 . on the basis of the results obtained from the mouse model , effective dosages of thymoquinone in humans were estimated using allometric scaling . according to the body weight ratio of man to mouse , a similar effect is to be expected in humans at dosages between 0 . 05 and 0 . 25 mg / kg . due to allometry , the effective dosages estimated for humans are smaller than that obtained for mice . 1 . cepinskas g , wilson j x . inflammatory response in microvascular endothelium in sepsis : role of oxidants . j clin biochem nutr 2008 ; 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