Cardiovascular diseases related to abnormal lipid metabolism are very frequent in industrialised countries. In Italy, for instance, according to the data from the World Health Organization published on April 2011, the number of decreases in Italy for cardiovascular diseases reached 18.65% of the overall mortality. Our knowledge of the relationships between cholesterol and coronary heart disease stem from epidemiological studies conducted over the past few years. The conclusions reached in these studies indicate that the development of severe coronary atherosclerosis and coronary heart disease are closely correlated with serum cholesterol levels (Breuer, H. W. M.; European Cardiology, 2005; 1-6).
Platelets play an important, but often under-recognized role in cardiovascular disease. For example, the normal response of the platelet can be altered, either by increased pro-aggregatory stimuli or by diminished anti-aggregatory substances to produce conditions of increased platelet activation/aggregation occurring in cardiovascular disease states both on a chronic (e.g. stable angina pectoris) and acute basis (e.g. acute myocardial infarction). In addition, platelet hyperaggregability is also associated with risk factors of coronary artery disease (e.g. smoking, hypertension, and hypercholesterolemia). Finally, the utility of an increasing range of anti-platelet therapies in the management of the above disease states further emphasizes the pivotal role platelets play in the pathogenesis of cardiovascular disease. A recently published paper provides a comprehensive overview of the normal physiologic role of platelets in maintain homeostasis, the pathophysiologic processes that contribute to platelet dysfunction in cardiovascular disease and the associated role and benefits of anti-platelet therapies (Kottke-Marchant K.: Cleveland Clinic Journal of Medicine; 2009 April; 76(1): 1-7).
Evidence is accumulating that most of the degenerative diseases that afflict humanity have their origin in deleterious free radical reactions. These diseases include atherosclerosis, cancer, inflammatory joint disease, asthma, diabetes, senile dementia and degenerative eye disease. The process of biological ageing might also have a free radical basis. Most free radical damage to cells involves oxygen free radicals or, more generally, activated oxygen species (AOS) which include non-radical species such as singlet oxygen and hydrogen peroxide as well as free radicals. The AOS can damage genetic material, cause lipid peroxidation in cell membranes, and inactivate membrane-bound enzymes. Antioxidant supplementation of our diet is needed to ensure a more healthy elderly population (Aust N Z J Ophthalmol. 1995 February; 23(1):3-7).
Omega-3 polyunsaturated fatty acids (n-3 PUFA) have demonstrated a beneficial effect in the prevention of cardiovascular events (Aarsetoey H. et al.; Cardiology Research and Practice, Volume 2012: 1-16), possibly by means of an antiinflammatory, antithrombotic and antiarrhythmic mechanism (Sethi S. et al.; Blood 2002:100:-1340-6; Billman G E, et al.; Circulation 3 1999: 99:2452-7). The hypolipidic effect was the first detected, so at first these drugs had been used for the treatment of dislipidemic disorders, while the antiinflammatory, antithrombotic, antiatherosclerotic and antiarrhythmogenic effects have been found later. GISSI-Prevention trial (Lancet 1999 354: 447-55) was the first trial demonstrating the efficacy and tolerability of n-3 PUFAs in post-myocardial infarction patients. According to the evidence in literature, today n-3 PUFAs are indicated for the primary and secondary prevention of ischemic cardiopathy and sudden cardiac death (SCD) (Mori T A, Beilin L J. Long-chain omega-3 fatty acids, blood lipids and cardiovascular risk reduction. Curr. Opin. Lipidol. 2001; 12:11-7). In Nutrition and Dietary Supplements 2011 September 14; 93-100 it is described the role of n-3 series polyunsaturated fatty acids in cardiovascular disease prevention.
Resveratrol (trans-3,4′,5,-trihydroxystilbene) is a polyphenol molecule located in the skins of black grapes. It is known that it has cardioprotective effects, acting as inhibitor of platelet aggregation (Szmitko P E, et al. Circulation January 2005, 111 (2) p 10-11; Das D K, et al. “Resveratrol in cardioprotection: a therapeutic promise of alternative medicine”; Mol. Interv., 2006, 6 (1): 36-47). It also acts as antioxidant and skin protecting agent (Afaq, Farrukh et al. “Botanical antioxidants in the prevention of photocarcinogenesis and photoaging”; Experimental Dermatology, 2006, 15 (9): 678-84). Resveratrol has been intensively studied recently, in relation to the known beneficial properties of red wine, of which it is one of the fundamental ingredients (Life Sci., 71, 2145-52, 2002). Numerous studies have demonstrated an anticarcinogenic activity of resveratrol, whose mechanisms of action of which can be subdivided as follows: inhibition of activation of transcription factor NF-kB, capable of regulating the expression of various genes involved in inflammatory and carcinogenic processes (Lancet, 341, 1103-1104, 1993; Science, 275, 218.220, 1997; Proc. Natl. Acad. Sc., 94, 14138-14143, 1997; Life Science, 61, 2103-2110, 1997; Brit. J. Pharm., 126, 673-680, 1999; J. Imm., 164, 6509-6519, 2000); inhibition of various proteins, including protein kinase C (Stewart, J. R., Ward, N. E., loannides, C. G. and O'Brian, C. A., Resveratrol preferentially inhibits protein kinase C-catalyzed phosphorylation of a cofactor-independent, arginine-rich protein substrate by a novel mechanism, Biochemistry. 1999, 38, 13244-13251), ribonucleotide reductase (FEBS Lett., 421, 277-279, 1998) and cyclo-oxygenase-2 (COX-2) in mammalian epithelial cells (Steinmetz K L, Tyson C K, Meierhenry E F, Spalding J W, Mirsalis J C. Examination of genotoxicity, toxicity and morphologic alterations in hepatocytes following in vivo or in vitro exposure to methapyrilene. Carcinogenesis. 1988 June; 9(6):959-963)); activation of caspases 2, 3, 6 and 9 (FASEB J., 1613-1615, 2000) and modulation of the gene p53, which is a known tumour suppressor (Soleas G J, Goldberg D M, Grass L, Levesque M, Diamandis E P. Do wine polyphenols modulate p53 gene expression in human cancer cell lines? Clin Biochem 2001; 34: 415-420).
In Free Radic. Res., 33, 105-114, 2000 it is described the antioxidant activity of resveratrol and its ability to counteract the damaging effects produced by various substances and/or conditions that cause intracellular oxidative stress.
In EP1567137B1 it is described the use of resveratrol for treating influenza virus infections.
WO 2011161501 describes solid compositions in form of powders or granulates wherein the active ingredient is insoluble or poorly soluble in water and/or thermolabile and/or having unpleasant organoleptic properties. The method described is a dispersion in a lipid matrix containing a triglyceride, a polyoxyethylene sorbitan ester and ascorbyl palmitate.
Patent application WO 2011120530 discloses solid porous inert carrier compositions comprising a porous silicium dioxide (silicon dioxide) and a release enhancing agent that can be loaded with pharmaceutical oils, among which fish oil. An example of preparation of compositions comprising n-3 PUFA in combination with one or more active ingredients is described in EP2517697. In particular, are described microcapsule suspensions comprising one or more statins in alkyl esters of n-3 PUFA, in which the statins are isolated from contact with the alkyl ester of n-3 PUFA by means of a polymeric membrane that can be easily disintegrated in the gastrointestinal medium.
Most of the methods known in the art, useful for preparing compositions containing resveratrol and n-3 PUFA, include microencapsulation or coating processes which are long and expensive. These studies are focused on finding suitable delivery methods of resveratrol in a single dose within an oily phase.
In the art methods for the solubilization of resveratrol in n-3 PUFA are not known.
Resveratrol, in fact, while keeping very good qualities as stabilizing agent in the prevention or reduction of the n-3 PUFA degradation, is almost insoluble in most of the oil phases used in the protocols for the preparation of medicines and nutraceuticals. Even the addition of certain emulsifiers and/or cosurfactant is not effective in achieving the desired purpose.
Considering the advantages of resveratrol in combination with n-3 PUFA fatty acids for medical or nutraceutical use, it becomes more and more noticeable the need to find an efficient method for solubilizing resveratrol in the oil phase.