MXPA06011060A - Use of a serine palmitoyltransferase (spt) inhibitor to treat atherosclerosis and dyslipidemia - Google Patents

Abstract

The present invention relates to methods of treating atherosclerosis, dyslipidemia, other cardiovascular diseases and related diseases, such as diabetes, using a serine palmitoyltransferase (SPT) inhibitor. The present invention also relates to pharmaceutical compositions and kits that comprise a serine palmitoyltransferase (SPT) inhibitor, optionally with another pharmaceutical agent.

Description

USE OF A SERINE PALM1TOIL TRANSFERASE (SPT) INHIBITOR TO TREAT ATEROSCLEROSIS AND DISLIPIDEMIA Field of the Invention The present invention relates to methods of using a compound that is a serine palmitoyl transferase (SPT) inhibitor to elevate certain levels of plasma lipids, including high density lipoprotein (HDL) cholesterol, and lowering other plasma lipid levels such as low density lipoprotein (LDL) cholesterol and triglycerides, and consequently, to treat diseases affected by low levels of HDL cholesterol and / or high levels of LDL cholesterol and triglycerides, such as atherosclerosis, dyslipidemia, hypercholesterolemia, hypertriglyceridemia, cardiovascular diseases and related diseases such as diabetes. The present invention also relates to pharmaceutical compositions and kits comprising an SPT inhibitor and a second therapeutic agent. Background of the Invention Atherosclerosis and its associated coronary artery disease (CAD) is the leading cause of mortality in the industrialized world. Despite attempts to modify secondary risk factors (for example, tobacco use, obesity, lack of exercise) and treatment of dyslipidemia with modifications in diet and drug therapy, coronary heart disease (CHD) remains the same. most common cause of death in the United States, where cardiovascular disease accounts for 44% of deaths, with 53% of these associated with coronary atherosclerotic heart disease. The pathological sequence that leads to atherosclerosis and coronary heart disease is well known. The earliest stage in this sequence is the formation of "lipid striae" in the carotid, coronary, and cerebral arteries and in the aorta. These lesions are yellow due to the presence of lipid deposits found mainly within smooth muscle cells and in macrophages of the intima of the arteries and the aorta. In addition, it has been postulated that the majority of the cholesterol found within the lipid striae, in turn, gives rise to the development of "fibrous plaques", which are composed of accumulated cells of smooth muscle of the intimate layer, loaded with lipids and surrounded by extracellular lipids, collagen, elastin and proteoglycans. The cells plus the matrix form a fibrous plug that covers a deeper deposit of cellular debris and more extracellular lipids. Lipids are mainly free cholesterol and esterified. The fibrous plaque forms slowly, and is likely to calcify and necrose over time, progressing to a "complicated lesion", which involves arterial occlusion and the tendency toward mural thrombosis and muscle spasms of the arterial walls that characterize atherosclerosis advanced The risk of the development of atherosclerosis and related cardiovascular disease has been shown to be strongly correlated with certain levels of plasma lipids. In recent years, the leaders of the medical profession have placed renewed emphasis on lowering plasma cholesterol levels, and low-density lipoprotein (LDL) cholesterol, in particular. It is known that levels above "normal" are significantly lower than those perceived up to now. As a result, large segments of the Western population now realize that they are at particularly high risk. These independent risk factors include glucose intolerance, hypertrophy of the left ventricle, hypertension, and belonging to the male sex. Cardiovascular disease is especially widespread among diabetic subjects, at least in part because of the existence of multiple independent risk factors in this population. The successful treatment of hyperlipidemia in the general population, and in diabetic subjects in particular, is therefore of exceptional medical importance. Although elevated LDL cholesterol may be the most recognized form of dyslipidemia, is by no means the only significant contributor to CHD associated with lipids. Low HDL-C is also a known risk factor for CHD (D.J. Gordon et al., "High-density Lipoprotein Cholesterol and Cardiovascular Disease", Circulation (1989) 79: 8-15). High levels of LDL cholesterol and triglycerides correlate positively, while high levels of HDL cholesterol correlate negatively with the risk of developing cardiovascular diseases. Therefore, dyslipidemia is not a unit risk profile of CHD but may be constituted by one or more lipid aberrations. There are no completely satisfactory lipid modulation therapies.

Niacin can significantly increase HDL cholesterol, but it has serious tolerance problems, which reduces compliance. Fibrates and HMG-CoA reductase inhibitors lower LDL cholesterol but raise HDL cholesterol only modestly (-10-12%). As a result, there is a significant unmet medical need for a well-tolerated agent, which can lower plasma LDL levels and / or raise plasma HDL levels (i.e., improve the patient's plasma lipid profile), this mode reversing or slowing the progression of atherosclerosis. Therefore, although there is a diversity of anti-atherosclerosis therapies, there is a continuing need and a continuing search for alternative therapies for the treatment of atherosclerosis and dyslipidemia. Serine palmitoyl transferase (SPT) catalyses the first stage involved in the synthesis of sphingolipids (Figure 1). SPT condenses palmitic acid from palmitoyl-coenzyme A with serine to produce cephalosphinganine, the initial precursor of the unique aminolipid structure that is characteristic of all sphingolipids (K. Hanada et al., J. Biol.Chem. 1997; 272 ( 51): 32108-14). The SPT is composed of two different subunits, LCB1 and LCB2 (B. Weiss and W. Stoffel, Eur.J.Biochem, 1997, 249 (1): 239-47, see also WO 99/49021). The LCB1 and LCB2 genes are essential for cell survival and changes in SPT activity result in defective development of the fruit fly and filamentous fungi (J. Cheng et al., Mol. Cell. Biol. 2001; 21 (18): 6198-209; and T. Adachi-Yamada et al., Mol. Cell. Biol. 1999; 19 (10): 7276-86), and hereditary sensory neuropathy type I in humans ( JL Dawkins et al., Nat. Genet.2001; 27 (3): 309-12, and K. Bejaoui et al., Nat. Genet., 2001; 27 (3): 261-2). Sphingomyelin is one of the main phospholipids in plasma lipoproteins and cell membranes. In vitro studies have shown that sphingomyelin and related sphingolipids are proatherogenic in a variety of circumstances and a positive correlation has been identified between the content of sphingomyelin (SM) in plasma and the incidence of coronary artery disease (X. Jiang et al. ., Arterioscler.Thromb.Vasc.Biol., 2000; 20: 2614-2618, and RD Williams, et al., J. Lipid Res. 1986. 27: 763-770). SM and its derivatives accumulate in human and experimental atherosclerotic lesions (S.L. Schissel et al., J Clin Invest., 1996; 98 (6): 1455-64). The intermediates of the synthesis of SM, in particular, ceramide, also have independent propriogenic properties. Ceramide plays an important role in the aggregation of lipoproteins and can promote foam cell formation (K. J. Williams and I. Tabas, Arterioscler, Thromb. Vasc. Biol. 1995; 15: 551-561). Although direct mechanistic links between MS and atherosclerosis have not been established, available in vitro data suggest that SM may have the following proatherogenic properties. First, the increased SM content of HDL and triglyceride-rich lipoproteins, for example, has been shown to hamper reverse cholesterol transport and the elimination of triglyceride-rich lipoproteins by interfering with lecithin: cholesterol acyltransferase (LCAT) activities (DJ Bolin et al. A. Jonas, J. Biol. Chem. 1996; 271 (32): 19152-8) and lipoprotein lipase (LPL) (I. Arimoto et al., J. Lipid Res. 1998; 39 (1): 143-51; I. Arimoto et al., Lipids 33: 773-779 (1996); and H. Saito et al., Biochimica et Biophysica Acta 1486 (2000) 312-320), respectively. It has also been shown that SM in macrophage membranes interferes with the reverse transport of cholesterol (A.R. Leventhal et al., J.Biol.Chem., 2001; 276 (48): 44976-83). Second, the SM-rich lipoproteins can be converted into foam cell substrates by sphingomyelinase in the arterial wall (SL Schissel et al., J. Biol. Chem. 1998; 273 (5): 2738-46), thereby promoting the formation of foam cells. Third, ceramide and the related products of synthesis and decomposition of SM are potent regulators of cell proliferation, activation and apoptosis (M. Maceyka et al., Biochim, Biophys, Acta. 2002; 1585 (2-3): 193-201) and therefore may affect plate growth and stability. Other proatherogenic effects of sphingolipids include the observation that SM in LDL enhances the reactivity of LDL with sphingomyelinase, which is released by macrophages in the arterial wall (Ts Jeong et al., J. CI in.Invest, 1998; 101 (4) : 905-912). This process results in the aggregation of LDL and the subsequent formation of foam cells (S.L. Schissel et al., J.Clin.Invest. 1996; 98 (6): 1455-1464). It is also known that the increased content of sphingomlelin in plasma membranes reduces the reverse cholesterol transport, hindering the transfer of cellular cholesterol to HDL (R. Kronqvist et al., Eur.J.Biochem, 1999; 262: 939-946). In addition, SPT activation is strongly implicated in Fas-mediated apoptosis, which could promote plate destabilization. Activation of Fas causes apoptosis in macrophages (PM Yao and I. Tabas, J.Biol.Chem., 2000; 275: 23807-23813) and smooth muscle cells (AC Knapp et al., Athero., 2000; 152: 217- 227). Activation of Fas depends on the de novo synthesis of ceramide, a SPT product and a precursor of SM (A. Cremesti et al., J.Biol.Chem, 2001; 276: 23954-23961). The genes that regulate cholesterol synthesis contain sterols regulatory elements (SREs) in their promoter regions (J.D. Horton, J.L. Goldstein and M.S. Brown, J. Clin.Research 2002; 109 (9): 1125-31). Through several intermediate steps, SREs are controlled by intracellular free cholesterol (M.S. Brown and J.L. Goldstein, Cell., 1997; 89 (3): 331-40). SM, a major component of the plasma membrane, has a high affinity for free cholesterol (TS Worgall et al., J. Biol. Chem. 200; 277 (6): 3878-85; and V. Puri et al., J. Biol. Chem. 2003; 278 (23): 20961-70). It has been reported that the depletion of MS by treatment with sphingomyelinase causes an increased translocation of cholesterol to the endoplasmic reticulum and the suppression of cleavage of SREBP (binding proteins to sterols regulatory elements) (S. Sheek, MS Brown and JL Goldstein, Proc. Nati, Acad. Sci. United States 1997; 94 (21): 11179-83). Recent discoveries showed that the inhibition of sphingolipid biosynthesis causes suppression of the expression of lipogenic genes in cells of the Chinese Hamster ovary (T.S. Worgall et al., Arterioscler, Thromb. Vasc. Biol. 2004; 24: 943-948). It is known that SPT inhibitors block the production of ceramide and the resulting apoptosis in cardiomyocytes (D. Dyntar et al., Diabetes 2001; 50: 2105-2113) and pancreatic insulin-producing β cells (M. Shimabukuro et al. , Proc.Natl.Acad: Sci. 1998; 95 (5): 2498-2502). Inhibition of SPT prevents apoptosis of pre-diabetic fa / fa rat islets (M. Shimabukuro et al., J.Biol.Chem 1998, 273 (49): 32487-90). Recent discoveries also showed that palmitate inhibits the expression of preproinsulin genes by ceramide biosynthesis. The inhibition of SPT recovered the expression of preproinsulin in rat islet culture and improved insulin production (C.L. Kelpe et al., J.Biol.Chem., 2003; 278 (32): 30015-21). Mithiocin is a known inhibitor of serine palmitoyl transferase (SPT) (K. Hanada et al., Biochem.Pharmacol., 2000; 59: 1211-1216; and JK Chen et al., Chemistry &Biology 1999; 6: 221- 235) isolated from fungi (Y. Miyake et al., Biochem. Biophys. Res. Commun. 1995; 211 (2): 396-403), which is commercially available, and is known to have potent immunosuppressive activity ( T. Fujita et al., J. Antibiot. (Tokyo) 1994; 47 (2): 208-15). Myroxine has been shown to possess immunomodulatory properties independent of its ability to inhibit SPT and by inhibiting growth in T-lymphocytes. WO 01/80903 describes the detection and treatment of atherosclerosis based on plasma sphingomyelin concentration. The documents WO 02/074924 and U.S. 2002/0197654, Thromb.

Haemost., 2001; 86: 1320-1326; describe a. method for comparatively measuring the level of normal and hyperproliferative expression of serine palmitoyl transferase in a mammalian cell and uses thereof, such as detecting cancer or treating restenosis. The U.S. 2003/9996022 describes methods and compositions useful for treating or preventing cardiovascular or cerebrovascular disease through the use of agents that interfere with the production and / or biological activities of sphingolipids and their metabolites, particularly sphingosine (SPH) and sphingosine-1-phosphate (S-1-P). WO 01/80715 describes methods for identifying compounds useful for preventing acute clinical vascular events in a subject. U.S. Patent 6,613,322; the U.S. 2003/0026796 and WO 99/11283 disclose methods for treating a subject suffering from atherosclerotic vascular disease comprising administering to the subject an amount of a zinc sphingomyelinase inhibitor effective to decrease the extracellular activity of zinc sphingomyelinase in the subject. The document by Tae-Sik Park et al., Circulation. 2004; 110: 3465-3471, describes the reduction of atherogenesis in Knockout mice for Apo-E by inhibiting sphingomyelin synthesis. The document by M. Hojjatl et al., JBC Papers in Press, published on December 6, 2004, as Original Document M412348200, describes the effect of myocin on the metabolism of sphingolipids in plasma and atherosclerosis in mice deficient in apoE. Summary of the Invention The present invention provides the following therapeutic methods: methods for lowering plasma lipids, comprising administering a therapeutically effective amount of a serine palmitoyl transferase (SPT) inhibitor to a mammal in need thereof; methods for raising high density lipoprotein (HDL) particles, comprising administering a therapeutically effective amount of a serine palmltoyl transferase (SPT) inhibitor to a mammal in need thereof; methods for lowering very low density lipoprotein (VLDL) particles and low density lipoprotein (LDL) particles, comprising administering a therapeutically effective amount of a serine palmitoyl transferase (SPT) inhibitor to a mammal in need thereof; methods for lowering plasma triglyceride particles, comprising administering a therapeutically effective amount of a serine palmltoyl transferase (SPT) inhibitor to a mammal in need thereof; methods for lowering serum total cholesterol levels, comprising administering a therapeutically effective amount of a serine palmitoyl transferase (SPT) inhibitor to a mammal in need thereof; methods for improving the plasma lipid profile, comprising administering a therapeutically effective amount of a serine palmltoyl transferase (SPT) inhibitor to a mammal in need thereof; methods for inhibiting plaque formation, comprising administering a therapeutically effective amount of a serine palmitoyl transferase (SPT) inhibitor to a mammal in need thereof; methods for reducing plaque size, comprising administering a therapeutically effective amount of a serine palmitoyl transferase (SPT) inhibitor to a mammal in need thereof; methods for reducing the size of an atherosclerotic lesion, comprising administering a therapeutically effective amount of a serine palmitoyl transferase (SPT) inhibitor to a mammal in need thereof; methods for reducing the size of a macrophage foam cell, comprising administering a therapeutically effective amount of a serine palmitoyl transferase (SPT) inhibitor to a mammal in need thereof; methods for preventing plaque rupture, comprising administering a therapeutically effective amount of a serine palmitoyl transferase (SPT) inhibitor to a mammal in need thereof; methods for treating dyslipidemia, comprising administering a therapeutically effective amount of a serine palmitoyl transferase (SPT) inhibitor to a mammal in need thereof; methods for treating atherosclerosis, comprising administering a therapeutically effective amount of a serine palmitoyl transferase (SPT) inhibitor to a mammal in need thereof; methods for treating diabetes, comprising administering a therapeutically effective amount of a serine palmitoyl transferase (SPT) inhibitor to a mammal in need thereof; methods for treating metabolic syndrome, comprising administering a therapeutically effective amount of a serine palmitoyl transferase (SPT) inhibitor to a mammal in need thereof; and finally, methods for treating inflammation, comprising administering a therapeutically effective amount of a serine palmitoyl transferase (SPT) inhibitor to a mammal in need thereof. More particularly, the present invention provides methods such that the SPT inhibitor is myrocin. In addition, the present invention provides pharmaceutical compositions comprising: a) a compound that is an inhibitor of serine palmitoyl transferase (SPT); and b) a second compound useful for the treatment of atherosclerosis or dyslipidemia. More particularly, the present invention provides compositions such that the second compound is an HMG-CoA reductase inhibitor, an HMG-CoA synthase inhibitor, an inhibitor of the expression of the HMG-CoA reductase gene, an expression inhibitor. of the HMG-CoA synthase gene, a CETP inhibitor, a bile acid sequestrant, an inhibitor of cholesterol absorption, a cholesterol biosynthesis inhibitor, a squalene synthetase inhibitor, a fibrate, niacin, a niacin combination and lovastatin and an antioxidant. Even more particularly, the present invention provides such compositions in which the second compound is an inhibitor of HMG-CoA reductase. Still more particularly, the present invention provides such compositions in which the second compound is lovastatin, simvastatin, pravastatin, fluvastatin, atorvastatin, rivastatin, rosuvastatin or pitavastatin. The present invention also provides such compositions in which the second compound is a CETP inhibitor. More particularly, the present invention provides compositions such that the second compound is torcetrapib. The present invention also provides compositions in which the SPT inhibitor is myriocin. In addition, the present invention provides kits comprising: a) a serine palmltoyl transferase (SPT) inhibitor and a pharmaceutically acceptable excipient, carrier or diluent in a first unit dosage form; b) a second compound that is useful for the treatment of atherosclerosis or dyslipidemia and a pharmaceutically acceptable excipient, carrier or diluent in a second unit dosage form; and c) a means for containing the first and second unit dosage forms. More particularly, the present invention provides such kits in which the second compound is an inhibitor of HMG-CoA reductase, an inhibitor of HMG-CoA synthase, an inhibitor of the expression of the HMG-CoA reductase gene, an inhibitor of the expression of the HMG-CoA synthase gene, a CETP inhibitor, a bile acid sequestrant, an inhibitor of cholesterol absorption, an inhibitor of cholesterol biosynthesis, an inhibitor of squalene, a fibrate, niacin, a combination of niacin and lovastatin and an antioxidant; and a pharmaceutically acceptable excipient, vehicle or diluent in a second unit dosage form; wherein the amounts of first and second compounds result in a therapeutic effect. Even more particularly, the present invention provides such kits in which the second compound is an HMG-CoA reductase inhibitor. More particularly, the present invention provides such kits in which the second compound is lovastatin, simvastatin, pravastatin, fluvastatin, atorvastatin, rivastatin, rosuvastatin or pitavastatin. In addition, the present invention provides such kits in which the second compound is a CETP inhibitor. More particularly, the present invention provides such kits in which the second compound is torcetrapib. In addition, the present invention provides such kits in which the SPT inhibitor is myriocin. The present invention also provides the use of a serine palmitoyl transferase (SPT) inhibitor for the manufacture or preparation of a medicament for the treatment of a mammal in need thereof, as described above. As indicated above, in clinical studies, plasma sphingomyelin (SM) levels have been correlated with the occurrence of coronary heart disease, regardless of plasma cholesterol levels. Myrocin is a potent inhibitor of serine palmitoyl transferase (SPT), the rate-limiting enzyme in the biosynthesis of ceramide and sphingomyelin (SM). In the present invention, it has been discovered that the inhibition of de novo biosynthesis of SM, using myriocin, improves the lipid profile and reduces atherogenesis in the knockout (KO) mouse for ApoE. Therefore, the present invention is directed to the use of SPT inhibitors to treat atherosclerosis, dyslipidemia and related diseases.

BRIEF DESCRIPTION OF THE DRAWINGS The present invention is further described by the following non-limiting examples, which refer to the accompanying Figures 1-14, of which small details are given below. In the present invention, the inhibition of SPT has been evaluated by measuring sphingomyelin, ceramide or sphinganine in plasma and tissues as a biomarker of inhibition. The present studies refer to an effect of a specific and commercially available SPT inhibitor, myriocin, in the reduction of lipid level and in the prevention of atherosclerosis in the knockout (KO) mouse for ApoE, a model conducive to Atherosclerosis Figure 1. Sphingomyelin Biosynthetic Route. Serine palmitoyl transferase (SPT) is the. first stage limiting the speed of the biosynthesis of esflngolípidos. Myroiocin specifically inhibits the SPT reaction. Figures 1-A, 1-B, 1-C, 1-D and 1-E. Gene expression and enzymatic activity of SPT. Figure 2-2B. Effect of myriocin on the distribution of lipoproteins in the plasma of KO mice for ApoE fed a Western diet for 4 weeks. HDL-high density lipoproteins (Figure 2-B); LDL-low density lipoproteins (Figure 2-A); VLDL-very low density lipoproteins (Figure 2). The SPT inhibitor, myriocin, was administered to KO mice for ApoE fed Western diet, mixed with the diet for 4 weeks at doses of 0 (control), 0.1, 0.3, and 1.0 mg / kg / day. Myxiocin caused a dose-dependent rise in HDL cholesterol and reduced lipoproteins containing apoB, LDL and VLDL (Figure 2). Figures 3-3A. Total concentrations of cholesterol and triglycerides in the plasma of KO mice for ApoE fed a Western diet for 4 weeks in the presence of myriocin. The KO control mice for ApoE were fed only Western diet. Cholesterol (Figure 3A) and total triglycerides (Figure 3B) in plasma were also reduced by myriocin. Figures 4-4A. Effect of myocycin on sphingomyelin in plasma and liver in KO mice for ApoE fed a Western diet for 4 weeks. Sphingomyelin was analyzed by LC / MS (liquid chromatography coupled to mass spectrometry). In addition, plasma and liver sphingomyelin concentrations (a potential biomarker based on mechanism) were reduced in a dose-dependent manner. Figures 5-5A. Effect of myocin on the development of lesions in the femoral artery with cuff of KO mice for ApoE fed a Western diet for 4 weeks. Changes in the lipid profile were accompanied by a significant reduction of atherosclerotic lesions in the cuffed femoral artery model (Arteriosclerosis and Thrombosis, Vol. 13, 1874-1884, 1993). Serum amyloid A levels in plasma were also determined. The atherosclerotic lesions (black bars) and the size of macrophages (gray bars) in the femoral artery were quantified. Figures 6-6B. Effect of myriocin on the distribution of lipoproteins in plasma of KO mice for ApoE fed with a Western diet for 12 weeks. In a single dose study, the SPT inhibitor, myriocin (0.3 mg / kg) was administered to KO mice for ApoE fed Western diet mixed with the diet for 12 weeks. The cholesterol profile in lipoproteins was examined using plasma isolated by FPLC (fast protein liquid chromatography). Treatment with mycocin reduced VLDL and LDL cholesterol, and increased HDL, respectively (Figure 6, A and B) when compared to KO mice for ApoE fed Western diet. The lipoprotein cholesterol levels of KO for ApoE mice treated with myriocin were comparable to that of KO mice for ApoE fed normal feed. The C57BI / 6J control mice showed a very low total plasma cholesterol. Figures 7-7A. Effect of myriocin (0, 3 mg / kg, mixed with the diet) on the total cholesterol and triglyceride concentrations of KO mice for ApoE fed a Western diet for 12 weeks. Plasma total cholesterol levels of KO mice for ApoE fed Western diet plus myriocin were lowered when compared to the Western diet fed group (Figure 7). In addition, myocynin treatment lowered plasma triglyceride levels (Figure 7A). The plasma total cholesterol and triglyceride levels of KO for ApoE mice treated with myriocin were comparable to that of KO mice for ApoE fed normal feed. On the other hand, wild-type C57BI / 6J mice showed low levels of total cholesterol and triglycerides in plasma. Figure 8-B. Effect of myriocin (0.3 mg / kg, mixed with the diet) on sphingomyelin (SM) concentrations in the liver, plasma and aorta of KO ApoE mice fed a Western diet for 12 weeks. Treatment with mycocin reduced the accumulation of MS in the liver (Figure 8). The KO mice for ApoE fed with the Western diet showed the highest levels of SM in plasma. Treatment with mycocin lowered plasma SM in KO mice for ApoE fed Western diet (Figure 8A). Small differences were observed between the aortas of different treatments. However, there were statistically significant differences between KO mice for ApoE fed Western diet and C57BI / 6J control mice; myriocin decreased SM levels in the aorta (Figure 8B). Figures 9-9A. Effect of mlriocin (0.3 mg / kg, mixed with the diet) on sphinganine concentrations in liver and aorta in KO mice for ApoE fed a Western diet for 12 weeks. Sphinganine levels were significantly increased in KO mice for ApoE fed Western diet as well as in KO mice for ApoE fed normal feed compared to the C57BI / 6J control mice. Treatment with mycocin reduced liver sphincganin levels when compared to KO mice for ApoE fed Western diet (Figure 9). In the aorta, sphinganine levels in the KO mice for ApoE treated with myroxine, KO mice for ApoE fed with normal feed, and control mice C57BI / 6J were lower than in the KO mice for ApoE fed Western diet (Figure 9 A). Figure 10. Effect of myriocin on the deposition of lipids in the aortas of KO mice for ApoE fed with the Western diet. Staining with Oil Red O on surfaces of the aortas showed that myocycin treatment reduced the diffusion of the atherosclerotic lesion in KO mice for ApoE fed Western diet (Figure 10). Figure 11. Effect of myriocin (0.3 mg / kg, mixed with the diet) on the formation of the total macrophage area and lesion in the root of the aorta. Figure 12. Effect of myriocin (0.3 mg / kg, mixed with the diet) on the formation of the macrophage area and total lesion in the brachiocephalic artery. The KO mice for ApoE were fed a Western diet in the absence or presence of myriocin. The KO mice for ApoE and the C57BI / 6J control mice fed normal feed were sacrificed. The atherosclerotic lesion (black bars) and the size of macrophages (gray bars) were quantified in the cross sections of the aortic root and the brachiocephalic artery using Image Pro Plus (Figures 11-12). Figure 13-13A. Proportion of SM / PC and ceramide concentrations in plasma. Treatment with myobinocin reduced ceramide levels and was not associated with any change in the SM / PC ratio. Figure 14. Incorporation of T lymphocytes in the lesion of the aortic root.

The accumulation of T cells was not affected by treatment with myriocin. Detailed Description of the Invention The present invention relates to methods for treating atherosclerosis, dyslipidemia, other cardiovascular diseases and related diseases, such as diabetes, using a compound that is a serine palmitoyl transferase (SPT) inhibitor. In addition, the present invention provides pharmaceutical compositions and kits comprising a serine palmitoyl transferase (SPT) inhibitor. According to the present invention, atherosclerosis, dyslipidemia, other cardiovascular diseases and related diseases, such as diabetes, can be treated by administering to a patient having or at risk of having said diseases, a therapeutically effective amount of a serine palmitoyl transferase inhibitor ( SPT). As shown in the Examples below, it has been demonstrated in the present invention that the content and production of SM increased proportionally in plasma, liver and aorta of KO mice for ApoE fed Western diet compared to KO mice for ApoE fed with conventional feed and the C57BI / 6J control mice. Mycocin, a specific SPT inhibitor, inhibited Novo synthesis of MS in the liver and aorta; this was associated with reductions in ceramide and SM in plasma that were not accompanied by changes in the SM / PC ratio. The inhibition of SM synthesis led to the lowering of cholesterol and triglycerides in plasma. These changes were associated with drastic anti-atherosclerotic effects in vivo. The depletion of SM was also associated with an elevation of HDL. The in vitro data suggest that the increased content of SM in lipoproteins can inhibit key enzymes involved in the metabolism of lipoproteins. It has also been shown that SM in the macrophage membranes prevented the reverse transport of cholesterol. It is conceivable that the depletion of SM could lead to the activation of reverse cholesterol transport and contribute to the elevation of HDL cholesterol, which is consistent with the observations of the present invention. In the present invention, inhibition of SM synthesis has been shown to be associated with significant reductions in the formation of atherosclerotic lesions in KO mice for ApoE. Since the plaque formation in KO mice for ApoE is lipid-directed, the anti-atherogenic effects observed were probably indirect, due to the normalization of plasma lipids as a result of the inhibition of SM synthesis by the liver. However, local inhibition of SM production in the aorta has also been demonstrated. The KO mice for ApoE fed the Western diet, treated with myriocin, showed a plasma lipid profile similar to that of KO mice for ApoE fed conventional feed, but their lesions were significantly smaller. Taken together, these findings suggest that the anti-atherogenic effects of myocin could be attributed, in part, to the local inhibition of SPT in the arterial wall. Thus, inhibition of SPT by myroxin in KO mice for ApoE effectively inhibited SM synthesis, an effect that was associated with an improved plasma lipid profile, and significant anti-atherogenic activity. Consistent with these observations are clinical reports indicating that MS is an independent risk factor for coronary heart disease and a plasma marker of coronary artery disease. The present invention shows that SPT and potentially other key enzymes that regulate SM synthesis could represent a novel class of molecular targets for the prevention of dyslipidemia, atherosclerosis and related diseases. The term "therapeutically effective amount" means an amount of a compound or combination of compounds that treats a disease; improves, attenuates or eliminates one or more symptoms of a particular disease; or prevents or delays the onset of one or more symptoms of a disease. The term "patient" means animals, such as dogs, cats, cows, horses, sheep, geese and humans. Particularly preferred patients are mammals, including humans of both sexes. The term "pharmaceutically acceptable" means that the substance or composition must be compatible with the other ingredients of a formulation, and not injurious to the patient. The terms "treating", "treating" or "treatment" include preventive (for example, prophylactic) and palliative treatment. The term "serine palmitoyl transferase (SPT) inhibitor" means a compound or a pharmaceutically acceptable salt thereof, which inhibits or blocks the enzyme, serine palmltoyl transferase (SPT). It is also contemplated that any additional pharmaceutically active compound used in combination with serine palmltoyl transferase (SPT) inhibitor may be a pharmaceutically acceptable salt of the additional active compound. The term "SPT inhibitor" includes, for example, natural or synthetic amino acid polypeptides, proteins, small synthetic organic molecules, or deoxy- or ribo-nucleic acid sequences, which bind to serine palmitoyl transferase with an affinity of about 20. times or more, compared to other proteins or nucleic acids. For example, but not by way of limitation, polyclonal or monoclonal antibodies (including classical or phage display antibodies) induced against the protein serine palmitoyl transferase or a peptide fragment thereof or nucleic acid probes that hybridize with serine mRNA Palmitoyl transferase are suitable for use in the present invention. The term "selective" means that a ligand binds with a greater affinity to a particular receptor when compared to the binding affinity of the ligand with another receptor. Preferably, the binding affinity of the ligand for the first receptor is about 50% or greater than the binding affinity for the second receptor. More preferably, the binding affinity of the ligand to the first receptor is about 75% or greater than the binding affinity to the second receptor. Even more preferably, the binding affinity of the ligand to the first receptor is about 90% or greater than the binding affinity to the second receptor. Serine palmitoyl transferase (SPT) inhibitors can be identified, for example, by screening a library of compounds. Methods for identifying enzyme inhibitors are well known to those skilled in the art. Specific procedures that can be used to identify inhibitors of serine palmitoyl transferase (SPT) are presented in other publications, such as WO 01/80913; U.S. 2002/0197654; K. Hanada, T. Hará and M. Nishijima, J. Biol. Chem., March 24, 2000; 275 (12): 8409-15; and K. Gable et al., J. Biol. Chem., March 17, 2000; 275 (11): 7597-603; which are hereby incorporated by reference in this document. Novel inhibitors are discovered using methods that measure the enzymatic activity of serine palmitoyl transferase.

Examples of known serine palmitoyl transferase (SPT) inhibitors include myocin, which is commercially available, D-cycloserine, sphingofungin B, sphingofungin C and viridiofungins. Other SPT inhibitors will be well known to those skilled in the art, for example, those described in WO 01/80903, such as lipoxamycin and haloalanines (JK Chen, Chemistry &Biology, April 1999, Vol. 6: 221 -235; and US 2002/0197654). The term "pharmaceutically acceptable salts" includes the salts of compounds that are, within the scope of medical judgment, suitable for use with patients without excessive toxicity, irritation, allergic response and the like, which correspond to a reasonable benefit / risk ratio and effective for its intended use, as well as the zwitterionic forms of the compounds, where possible. The term "salts" refers to Inorganic and organic salts of compounds. These salts can be prepared in situ during the final isolation and purification of a compound, or by separately reacting a purified compound with a suitable organic or inorganic acid or base, as appropriate, and isolating the salt formed in this way. Representative salts include the salts hydrobromide, hydrochloride, sulfate, bisulfate, nitrate, acetate, oxalate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate, tosylate, besylate, esylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate. , mesylate, glucoheptonate, lactobionate, and laurylsulfonate, and the like. These may include cations based on alkalis and alkaline earth metals, such as sodium, lithium, potassium, calcium, magnesium, and the like, as well as non-toxic ammonium, quaternary ammonium, and amine cations including, but not limited to, ammonium, tetramethylammonium, tetraethylammonium , methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine and the like. See, for example, S.M. Berge, et al., "Pharmaceutical Salts", J. Pharm Sci, 66: 1-19 (1977). An inhibitor of serine palmitoyl transferase (SPT) can contain asymmetric or chiral centers, and therefore, exist in different stereoisomeric forms. It is contemplated that all stereoisomeric forms, as well as mixtures thereof, including racemic mixtures, form part of the present invention. In addition, the present invention contemplates all geometric and positional isomers. For example, if a compound contains a double bond, both cis and trans forms, as well as mixtures, are contemplated. Mixtures of isomers, including stereoisomers, can be separated into their individual isomers based on their physical and chemical differences by methods well known to those skilled in the art, such as chromatography and / or fractional crystallization. The enantiomers can be separated by converting the enantiomeric mixture into a diastereomeric mixture by reaction with an appropriate optically active compound (eg, alcohol), separating the diastereomers, and converting (eg, hydrolyzing) the individual diastereomers to the corresponding pure enantiomers. In addition, some of the compounds of this invention can be atropisomers (e.g., substituted biaryls) and are considered as part of this invention. An inhibitor of serine palmitoyl transferase (SPT) can exist in unsolvated as well as solvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like. The present invention contemplates and encompasses both solvated and unsolvated forms. It is also possible that a serine palmitoyl transferase (SPT) inhibitor may exist in different tautomeric forms. All tautomers of a serine palmitoyl transferase (SPT) inhibitor are contemplated. It is also intended that the invention described herein encompass compounds that are synthesized in vitro using laboratory techniques, such as those well known to synthetic chemists.; or synthesized using in vivo techniques, such as through metabolism, fermentation, digestion, and the like. It is also contemplated that the compounds can be synthesized using a combination of in vivo and in vitro techniques. The present invention also includes isotopically-labeled compounds, which are identical to the non-isotopically labeled compounds, except for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number. which is usually found more abundantly in nature. Examples of isotopes that can be incorporated into compounds identified by the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, such as 2H, 3H, 13C, 14C, 15N, 18O, 17O, 31P, 32P , 35S, 18F, 135L and 36CI, respectively. Inhibitors of SPT and pharmaceutically acceptable salts thereof, which contain the aforementioned isotopes and / or other isotopes of other atoms are within the scope of this invention. Certain isotopically-labeled compounds of the present invention, for example those in which radioactive isotopes such as 3H and 14C are incorporated, are useful in drug and / or tissue substrate distribution assays. The tritiated isotopes, i.e., 3H, and carbon-14, i.e., 14C, are particularly preferred for their ease of preparation and detectability. In addition, replacement with heavier isotopes such as deuterium, i.e., 2H, can produce certain therapeutic advantages as a result of increased metabolic stability, for example increased half-life in vivo or reduced dosage requirements and, thus, may be preferred. in some circumstances. Isotopically-labeled compounds can generally be prepared by substituting an isotopically non-labeled reagent for an isotopically readily available unlabeled reagent. Metabolic syndrome, also known as Syndrome X or insulin resistance, refers to a common clinical condition that is defined as the presence of increased insulin levels in conjunction with other disorders including visceral obesity, hyperlipidemia, dyslipidemia, hyperglycemia, hypertension, and potentially hyperuricemia and renal dysfunction. A serine palmitoyl transferase (SPT) inhibitor is administered to a patient in a therapeutically effective amount. A serine palmitoyl transferase (SPT) inhibitor can be administered alone or as part of a pharmaceutically acceptable composition. In addition, a compound or composition can be administered all at once, such as, for example, by bolus injection, several times, such as by a series of tablets, or delivered substantially uniformly over a period of time, as for example using transdermal delivery. It is also indicated that the dose of the compound can be modified over time. A serine palmitoyl transferase (SPT) inhibitor can be administered using an immediate release formulation, a controlled release formulation, or combinations thereof. The term "controlled release" includes sustained release, delayed release and combinations thereof. A serine palmitoyl transferase (SPT) inhibitor and other pharmaceutically active compounds, if desired, can be administered to a patient orally, rectally, parenterally (eg, intravenously, intramuscularly or subcutaneously). ), intracisternally, intravaginally, intraperitoneally, intravesicularly, locally (for example, powders, ointments, or drops), or in the form of a buccal or nasal spray. Compositions suitable for parenteral injection may comprise physiologically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions, emulsions or emulsions, or may comprise sterile powders for reconstitution into sterile injectable solutions or dispersions. Examples of excipients, diluents, solvents or suitable aqueous and non-aqueous vehicles include water, ethanol, polyols (propylene glycol, polyethylene glycol, glycerol, and the like), suitable mixtures thereof, triglycerides, including vegetable oils such as olive oil, or injectable organic esters such as ethyl oleate. A preferred excipient is Miglyol®. Proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersions and / or by the use of surfactants. These compositions may also contain adjuvants such as preservatives, moisturizers, emulsifiers, and / or dispersing agents. The pollution prevention of. the compositions by microorganisms can be made by the addition of various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, and the like. It may also be desirable to include isotonic agents, for example sugars, sodium chloride, and the like. Prolonged absorption of injectable pharmaceutical compositions can be brought about by the use of agents capable of delaying absorption, for example, aluminum monostearate, and / or gelatin. Solid dosage forms for oral administration include capsules, tablets, powders, and granules. In said solid dosage forms, the active compound is mixed with at least one usual inert excipient (or vehicle), such as sodium citrate or dicalcium phosphate or (a) fillers or diluents, such as, for example, starches, lactose, sucrose, mannitol , or silicic acid; (b) binders, such as, for example, carboxymethyl cellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, or gum arabic; (c) humectants, such as, for example, glycerol; (d) disintegrating agents, such as for example, agar-agar, calcium carbonate, potato starch or tapioca, alginic acid, certain complex silicates or sodium carbonate; (e) solution retardants, such as paraffin; (f) absorption accelerators, such as, for example, quaternary ammonium compounds; (g) humidifying agents, such as, for example, cetyl alcohol or glycerol monostearate; (h) adsorbents, such as kaolin or bentonite; and / or (i) lubricants, such as, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules and tablets, the dosage forms may also comprise buffering agents. Solid compositions of a similar type can also be used as fillers in soft or hard filled gelatin capsules, using excipients such as lactose, milk sugar, as well as high molecular weight polyethylene glycols, and the like. Solid dosage forms such as tablets, dragees, capsules and granules can be prepared with coatings or shells, such as enteric coatings and others well known in the art. They may also contain opacifying agents, and may also be of such composition that they release the active compound or compounds in a delayed manner. Examples of embedded compositions that may be used are polymeric substances and waxes. The active compounds may also be in microencapsulated form, if appropriate, with one or more of the aforementioned excipients. Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs. In addition to the active compounds, the liquid dosage form may contain inert diluents, commonly used in the art, such as water or other solvents, solubilizing agents and emulsifiers, such as for example ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, ethyl, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils, in particular, cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, oil Sesame seeds, Miglyol®, glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols, sorbitan esters of fatty acids, or mixtures of these substances, and the like. In addition to said inert diluents, the composition may also include adjuvants, such as wetting agents, emulsifiers, and suspending agents, sweeteners, flavors, and perfuming agents.

The suspensions, in addition to the active compound, may contain suspending agents, such as for example ethoxylated isostearyl alcohols, polyoxyethylene sorbltol or sorbitan esters, microcrystalline cellulose, aluminum metahydroxyl, bentonite, agar-agar or tragacanth, or mixtures of these substances, and the like. . Compositions for rectal or vaginal administration can be prepared by mixing a serine palmitoyl transferase (SPT) inhibitor and any additional compounds with suitable non-irritating carriers or excipients, such as cocoa butter, polyethylene glycol or a suppository wax, which are solid at normal room temperature , but liquid at body temperature, and therefore, they melt in the rectum or in the vaginal cavity and release the compound. Dosage forms for topical administration of a serine palmltoyl transferase (SPT) inhibitor include ointments, powders, sprays and inhalers. The compound (s) is mixed under sterile conditions with a physiologically acceptable excipient, and any preservatives, buffers, and / or propellants that may be required. Ophthalmic formulations, eye ointments, powders, and solutions are also contemplated within the scope of this invention. A serine palmitoyl transferase (SPT) inhibitor can be administered to a patient at dosage levels in the range of about 0.1 to about 7,000 mg per day. A preferred dosage range is from about 1 to about 100 mg per day. The specific dosage and dosage range that can be used depend on several factors, including the requirements of the patient, the severity of the condition or disease being treated, and the pharmacological activity of the compound being administered. The determination of the dosing intervals and optimal dosages for a particular patient is within the usual capabilities of a person skilled in the art in view of this description. The present invention relates to the use of serine palmitoyl transferase (SPT) inhibitors to treat atherosclerosis, dyslipidemia and other cardiovascular diseases. The methods of treatment of the present invention may also include combination therapy, wherein other pharmaceutically active compounds useful for the treatment of atherosclerosis, dyslipidemia, or other cardiovascular diseases are used in combination with a serine palmitoyl transferase (SPT) inhibitor. In one embodiment of the present invention, a patient having or at risk of having atherosclerosis may be administered a combination of: 1) serine palmitoyl transferase (SPT) inhibitor; and 2) an additional compound useful for treating atherosclerosis, dyslipidemia, or other cardiovascular diseases, or combinations of compounds useful in treating these diseases. In addition, a serine palmitoyl transferase (SPT) inhibitor can be administered in combination with other pharmaceutical agents such as inhibitors of cholesterol biosynthesis and cholesterol absorption inhibitors, especially HMG-CoA reductase inhibitors and HMG-CoA synthase inhibitors, gene expression inhibitors of HMG-CoA reductase and synthase, CETP inhibitors, bile acid sequestrants, fibrates, inhibitors of ACAT (acyl-coenzyme A cholesterol acyltransferase), squalene synthetase inhibitors, antioxidants and niacin. An inhibitor of serine palmitoyl transferase (SPT) can also be administered in combination with naturally occurring compounds that act to lower plasma cholesterol levels. These naturally occurring compounds are usually referred to as nutraceuticals, and include, for example, garlic extract, Benecol®, and niacin. A slow-release form of niacin is available and is known as Niaspan. Niacin can also be combined with other therapeutic agents such as lovastatin, which is an inhibitor of HMG-CoA reductase and is described below. This combination therapy is known as ADVICOR ™ (Kos Pharmaceuticals Inc.). Any inhibitor of cholesterol absorption can be used as the second compound in the combination aspect of the present invention. The expression "inhibition of cholesterol absorption" refers to the ability of a compound to prevent cholesterol contained in the lumen of the intestine from entering the intestinal cells and / or passing from within the intestinal cells into the bloodstream. Said cholesterol absorption inhibiting activity is easily determined by those skilled in the art according to conventional assays (for example, J. Lipid Res. (1993) 34: 377-395). Inhibitors of cholesterol absorption are known to those skilled in the art and are described, for example, in PCT WO 94/00480. An example of a recently approved cholesterol absorption inhibitor is ZETIA ™ (ezetimibe) (Merck / Schering-Plow). Any HMG-CoA reductase inhibitor can be used as an additional compound in the combination therapy aspect of the present invention. The term "HMG-CoA reductase inhibitor" refers to a compound that inhibits the biotransformation of hydroxymethylglutaryl-coenzyme A to mevalonic acid catalyzed by the enzyme HMG-CoA reductase. Such inhibition can be readily determined by one skilled in the art according to conventional tests, (e.g., Methods of Enzymology, 71: 455-509 (1981); and the references cited in this document). A variety of these compounds are described and mentioned below. U.S. Patent No. 4,231,938 discloses certain compounds isolated after culturing a microorganism belonging to the Aspergillus genus, such as lovastatin. In addition, U.S. Patent No. 4,444,784 discloses synthetic derivatives of the aforementioned compounds, such as simvastatin. In addition, U.S. Patent No. 4,739,073 discloses certain substituted Intents, such as fluvastatin. In addition, U.S. Patent No. 4,346,227 describes derivatives of ML-236B, such as pravastatin. In addition, European Patent No. 491,226 shows certain pyridyldihydroxyheptenoic acids, such as rivastatin. In addition, U.S. Patent Nos. 4,681,893 and 5,273,995 disclose certain 6- [2- (substituted pyrrol-1-yl) -alkull] -pyran-2-ones, such as atorvastatin and the hemichaltion salt of the same (Lipltor®). Those skilled in the art will know of other HMG-CoA reductase inhibitors, such as rosuvastatin and pitavastatin. Examples of marketed products containing HMG-CoA reductase inhibitors that can be used in combination with compounds of the present invention include Baycol®, Lescol®, Lipitor®, Mevacor®, Pravachol® and Zocor®.

Any HMG-CoA synthase inhibitor can be used as the second compound in the combination therapy aspect of this invention. The term "HMG-CoA synthase inhibitor" refers to a compound that inhibits the biosynthesis of hydroxymethylglutaryl-coenzyme A from acetyl-coenzyme A and acetoacetyl-coenzyme A, catalyzed by the enzyme HMG-CoA synthase. Such inhibition can be readily determined by one skilled in the art in accordance with conventional assays (eg, Methods of Enzymology, 35: 155-160 (1975); and Methods of Enzymology, 110: 19-26 (1985); cited in this document). A variety of these compounds are described and mentioned below. U.S. Patent No. 5,120,729 discloses certain beta-lactam derivatives. U.S. Patent No. 5,064,856 discloses certain spiro-lactone derivatives prepared by culturing microorganism MF5253. U.S. Patent No. 4,847,271 discloses certain oxetane compounds, such as 11- (3-hydroxymethyl-4-oxo-2-oxetail) -3,5,7-trimethyl-2,4-undecadienoic acid derivatives . Those skilled in the art will know of other HMG-CoA synthase inhibitors. Any compound that decreases the expression of the HMG-CoA reductase gene can be used as an additional compound in the combination therapy aspect of this invention. These agents can be transcription inhibitors of HMG-CoA reductase that block transcription of DNA or translation inhibitors that prevent translation of the mRNA encoding the HMG-CoA reductase in the protein. Such inhibitors can directly affect transcription or translation, or they can be biotransformed into compounds having the aforementioned attributes by one or more enzymes of the cholesterol biosynthesis cascade, or they can lead to the accumulation of an isoprene metabolite having the above activities mentioned. Such regulation is readily determined by those skilled in the art in accordance with conventional assays (Methods of Enzymology, 110: 9-19 1985). Several such compounds are described and mentioned below, although other inhibitors of HMG-CoA reductase gene expression will be known to those skilled in the art. U.S. Patent No. 5,041,432 discloses certain substituted lanosterol derivatives. Other oxygenated stellates that suppress the biosynthesis of HMG-CoA reductase are described by E.l. Mercer (Prog. Lip. Res., 32: 357-416 1993). Any compound having activity as a CETP inhibitor can serve as a second compound in the combination therapy aspect of the present invention. The term "CETP inhibitor" refers to a compound that inhibits transport mediated by cholesteryl ester transfer protein (CETP) by transporting various cholesterol esters and triglycerides from HDL to LDL and VLDL. Such inhibition of CETP is readily determined by those skilled in the art in accordance with conventional tests (e.g., U.S. Patent No. 6,140,343). Those skilled in the art will know a variety of CETP inhibitors, for example, those described in the commonly assigned US Patent No. 6,140,343 and the commonly assigned United States Patent No. 6,197. .786. The CETP inhibitors described in these patents include compounds, such as [2R, 4S] 4 - [(3,5-bis-trifluoromethyl-benzyl) -methoxycarbonyl-amino] -2-ethyl-6-trifluoromethyl ethyl ester. -3,4- dihydro-2H-quinoline-1 -carboxylic acid, which is also known as torcetrapib. U.S. Patent No. 5,512,548 discloses certain polypeptide derivatives having activity as inhibitors of CETP, while certain CETP-inhibiting rosenonolactone derivatives, and phosphate-containing cholesteryl ester analogs are disclosed in J. Antibiot., 49 (8): 815-816 (1996), and Bioorg. Med. Chem. Lett.; 6: 1951-1954 (1996), respectively. Any ACAT inhibitor can serve as an additional compound in the combination therapy aspect of this invention. The term "ACAT inhibitor" refers to compounds that inhibit the intracellular esterification of dietary cholesterol by the enzyme acyl CoA: cholesterol acyltransferase. Such inhibition can be readily determined by one skilled in the art according to conventional tests, such as the procedure of Heider et al. described in Journal of Lipid Research., 24: 1127 (1983). A variety of these compounds are described and mentioned below. However, other ACAT inhibitors will be known to those skilled in the art. U.S. Patent No. 5,510,379 discloses certain carboxysulfonates, while WO 96/26948 and WO 96/10559 both disclose urea derivatives having ACAT inhibitory activity. Any compound that has activity as an inhibitor of squalene synthetase can serve as an additional compound in the aspect of combination therapy of the present invention. The expression "squalene synthetase inhibitor" refers to a compound that inhibits the condensation of two molecules of farnesylpyrrophosphate to form squalene, a reaction that is catalyzed by the enzyme squalene synthetase. Such inhibition is readily determined by those skilled in the art according to conventional methodology (Methods of Enzymology, 15: 393-454 (1969); and Methods of Enzymology, 110: 359-373 (1985); and references cited therein. document). A summary of squalene synthetase inhibitors has been compiled in Curr. Op. Ther. Patents, 861-4, (1993). Other compounds that are marketed for hyperlipidemia, including hypercholesterolemla and which are intended to help prevent or treat atherosclerosis include bile acid sequestrants, such as Welchol®, Colestld®, LoCholest® and Questran®; and fibric acid derivatives, such as Atromid®, Lopid®, and Tricor®. These compounds can also be used in combination with a serine palmitoyl transferase (SPT) inhibitor. The inhibition of SPT may be beneficial not only for atherosclerosis, but also for conditions such as type II diabetes, lipotoxicity and insulin sensitivity. It has been shown that chronic exposure to fatty acids due to obesity or hyperglycemia causes pancreatic β-cell apoptosis (lipotoxicity) and the interruption of the insulin response by ceramide generation (M. Shimabukuro et al., Proc Nati Acad Sci US A. 1998; 95: 2498-502). Diabetes can be treated by administering to a patient having diabetes (especially type II), insulin resistance, impaired tolerance to glucose, or the like, or any of the diabetic combinations such as neuropathy, nephropathy, retinopathy, or cataracts, a therapeutically amount effective of an SPT inhibitor in combination with other agents (e.g., insulin) that can be used to treat diabetes. This includes the classes of antidiabetic agents (and specific agents) described in this document. Any glycogen phosphorylase inhibitor can be used as the second agent in combination with an SPT inhibitor of the present invention. The expression "inhibitor of glycogen phosphorylase" refers to compounds that inhibit the bioconversion of glycogen to glucose-1-phosphate which is catalyzed by the enzyme glycogen phosphorylase. Said activity of glycogen phosphorylase inhibition is easily determined by those skilled in the art according to conventional tests (for example, J. Med. Chem. 41 (1998) 2934-2938). A variety of glycogen phosphorylase inhibitors are known to those skilled in the art, including those described in WO 96/39384 and WO 96/39385. Any aldose reductase inhibitor can be used in combination with an SPT inhibitor of the present invention. The expression "aldose reductase inhibitor", refers to compounds that inhibit the bioconversion of glucose to sorbitol, which is catalyzed by the enzyme aldose reductase. The inhibition of aldose reductase is readily determined by those skilled in the art using conventional assays (eg, J. Malone, Diabetes, 29: 861-864 (1980). "Red Cell Sorbitol, and Indicator of Diabetic Control"). A variety of aldose reductase inhibitors are known to those skilled in the art. Any sorbitol dehydrogenase inhibitor can be used in combination with an SPT inhibitor of the present invention. The term "sorbitol dehydrogenase inhibitor" refers to compounds that inhibit the bioconversion of sorbitol to fructose which is catalyzed by the enzyme sorbitol dehydrogenase. Said sorbitol dehydrogenase inhibition activity is easily determined by those skilled in the art according to conventional tests (eg, Analyt, Biochem (2000) 280: 329-331). A variety of sorbitol dehydrogenase inhibitors are known, for example, U.S. Patent Nos. 5,728,704 and 5,866,578 disclose compounds and a method for treating or preventing diabetic complications by inhibiting the enzyme sorbitol dehydrogenase. Any glucosidase inhibitor can be used in combination with an SPT inhibitor of the present invention. A glucosidase inhibitor inhibits the enzymatic hydrolysis of complex carbohydrates by glycoside hydrolases, for example, amylase or maltase, in simple, bioavailable sugars, eg, glucose. The rapid metabolic action of the glucosidases, particularly after the intake of high levels of carbohydrates, results in a state of alimentary hyperglycemia, which in diabetic or adipose subjects, leads to an enhanced secretion of insulin, increased synthesis of fats and a reduction in the degradation of fats. After such hyperglycemia, hypoglycaemia frequently occurs due to the increased levels of insulin present. In addition, it is known that the chyme that remains in the stomach promotes the production of gastric juices, which initiate or favor the development of gastritis or duodenal ulcers. Accordingly, it is known that glucosidase inhibitors are useful in accelerating the passage of carbohydrates through the stomach and inhibiting the absorption of glucose from the intestine. In addition, the conversion of carbohydrates to fatty tissue lipids and the subsequent incorporation of dietary fats into fatty tissue deposits is reduced or delayed accordingly, with the concomitant benefit of reducing or avoiding the damaging abnormalities resulting therefrom. Said glucosidase inhibition activity is easily determined by those skilled in the art according to conventional tests, (e.g., Biochemistry (1969) 8: 4214). A generally preferred glucosidase inhibitor includes an amylase inhibitor. An amylase inhibitor is a glucosidase inhibitor that inhibits the enzymatic degradation of starch or glycogen to maltose. Said amylase inhibition activity is easily determined by those skilled in the art in accordance with conventional assays (eg, Methods Enzymol. (1955) 1: 149). The inhibition of said enzymatic degradation is beneficial in reducing the amounts of bioavailable sugars, including glucose and maltose, and the attendant concomitant conditions resulting therefrom. A variety of glucosldase inhibitors are known to one skilled in the art, and examples are given below. Preferred glucosidase inhibitors are the inhibitors which are selected from the group consisting of acarbose, adiposine, voglibose, miglltol, emiglitate, camiglibose, tendamistate, trestatin, pradimicin-Q and salbostatin. The glucosidase inhibitor, acarbose, and the various amino sugar derivatives related thereto are described in U.S. Patent Nos. 4,062,950 and 4,174,439 respectively. The glucosidase inhibitor, adiposine, is described in U.S. Patent No. 4,254,256. The glucosidase inhibitor, vogllbosa, 3,4-dideoxy-4 - [[2-hydroxy-1- (hydroxymethyl) etyl] amino] -2-C- (hydroxymethyl) -D-epi-inositol, and the various pseudoaminoazúcares N Substitutes related thereto are described in U.S. Patent No. 4,701,559. The glucosidase inhibitor, miglitol, (2 /? 3 /? 4 5S) -1- (2-hydroxyethyl) -2- (hydroxymethyl) -3,4,5-piperidinetriol, and the various 3,4,5 -trihydroxypiperidines related thereto are described in U.S. Patent No. 4,639,436. The glucosidase inhibitor, emiglltate, p- [2 - [(2 3R, 4 5S) -3,4,5-trihydroxy-2- (hydroxymethyl) piperidino] ethoxy] ethyl benzoate, the various derivatives related thereto and pharmaceutically acceptable acid addition salts thereof, are described in U.S. Patent No. 5,192,772. The glucosidase inhibitor, MDL-25637, 2,6-dideoxy-7-O-β-D-glucopyranosyl-2,6-imino-D-glycero-L-glucoheptol, the various homodsaccharides related thereto and the pharmaceutically acceptable acid addition salts thereof are described in U.S. Patent No. 4,634,765. The glucosidase inhibitor, camiglibose, 6-deoxy-6 - [(2R, 3R, 4f?, 5S) -3,4,5-trihydroxy-2- (hydroxymethyl) piperidino] -D-glucopyranoside sesquihydrate , the deoxynojirimycin derivatives related thereto, the various pharmaceutically acceptable salts thereof, and synthetic methods for the preparation thereof, are disclosed in U.S. Patent Nos. 5,157,116 and 5,504,078. The glucosidase inhibitor, salbostatin, and the various pseudosaccharides related thereto, are described in U.S. Patent No. 5,091,524. A variety of amylase inhibitors are known to a person skilled in the art. The tendamistat amylase inhibitor and the various cyclic peptides related thereto are described in U.S. Patent No. 4,451,455. The amylase inhibitor AI-3688 and the various cyclic polypeptides related thereto are described in U.S. Patent No. 4,623,714. The amylase inhibitor, trestatin, constituted by a mixture of trestatin A, trestatin B and trestatin C, and the various trehalose-containing amino sugars related thereto are disclosed in U.S. Patent No. 4,273,765. Additional anti-diabetic compounds, which can be used as the second agent in combination with an SPT inhibitor of the present invention include, for example, the following: biguanides (e.g., metformin), insulin secretagogues (e.g., sulfonylureas and glinides) ), glitazones, PPAR agonists? no glitazone, PPARβ agonists, DPP-IV inhibitors, PED5 inhibitors, GSK-3 inhibitors, glucagon antagonists, f-1-6-BPase inhibitors (metabasis / Sankyo), GLP-1 analogues (AC 2993 , also known as exendin-4), insulin and insulin mimics (Merck natural products). Other examples could include inhibitors of PKC-ß and AGE breakers. As described above, a serine palmitoyl transferase (SPT) inhibitor can be administered alone or with other pharmaceutically active compounds. The other pharmaceutically active compounds may claim to treat the same disease as the serine palmitoyl transferase inhibitor (SPT) or a different disease. If the patient is going to receive or receive multiple pharmaceutically active compounds, the compounds can be administered simultaneously or sequentially in any order. For example, in the case of tablets, the active compounds can be in a tablet or in different tablets, which can be administered at the same time or sequentially in any order. In addition, it must be admitted that the composition can be of different forms. For example, one or more. Compounds can be supplied by one tablet, while another is administered by injection or orally in the form of syrup. All combinations, supply procedures and administration sequences are contemplated. Since one aspect of the present invention contemplates the treatment of the aforementioned diseases with a combination of pharmaceutically active agents that can be administered separately, the invention further relates to combining different pharmaceutical compositions in the form of a kit. For example, a kit can comprise two different pharmaceutical compositions comprising: 1) an inhibitor of serine palmitoyl transferase (SPT); and 2) a second pharmaceutically active compound. The kit also comprises a container for the different compositions, such as a divided bottle, or a package of divided aluminum foil. Additional examples of containers include syringes, boxes, bags, and the like. Typically, a kit comprises instructions for the administration of the different components. The kit form is particularly advantageous when the different components are preferably administered in different dosage forms (eg, oral and parenteral) are administered in different dosage ranges, or when the titration of the individual components of the combination is desired by the Prescribing doctor. An example of a klt is a blister pack. Blister packs are well known in the packaging industry and are widely used for the packaging of dosage unit dosage forms (tablets, capsules, and the like). The blister packs are generally constituted by a sheet of relatively rigid material coated with a sheet of preferably transparent plastic material. During the packaging process, gaps are formed in the plastic sheet. The holes have the size and shape of the tablets or capsules to be packaged. Then, the tablets or capsules are placed in the recesses and a sheet of relatively rigid material is sealed against the plastic sheet by the side of the sheet opposing the direction in which the recesses were formed. As a result, the tablets or capsules are hermetically sealed in the recesses between the paper, the plastic sheet and the sheet. Preferably, the strength of the sheet is such that the tablets or capsules can be removed from the blister pack by manually applying pressure to the voids to thereby form an opening in the sheet at the location of the pocket. The tablet or capsule can then be withdrawn through said opening. It may be desirable to provide a reminder in the kit, for example, in the form of numbers close to the tablets or capsules, in which the numbers correspond to the days of the regimen in which the specified tablets or capsules are to be ingested. Another example of this reminder is a calendar printed on the card, for example, as follows: "First Week, Monday, Tuesday, ... etc ... Second Week, Monday, Tuesday," etc. Other variants of reminders will be readily apparent. A "daily dose" can be a single tablet or capsule or several pills or capsules to be taken on a given day. In addition, a daily dose of a serine palmitoyl transferase inhibitor (SPT) can be constituted by a tablet or capsule, while a dose of the second compound can be constituted by several tablets or capsules and vice versa. The reminder should remember this and help the correct administration of the active agents. In another embodiment of the present invention, a dispenser designed to dispense the daily doses one at a time in the order of its intended use is provided. Preferably, the dispenser is equipped with a reminder, in order to further facilitate compliance with the dosing regimen. An example of such a reminder is a mechanical counter, which indicates the number of daily doses that have been dispensed. Another example of such a reminder is a battery-operated memory chip associated with a liquid crystal reader, or an audible warning signal which, for example, reads aloud the date on which the last daily dose was taken and / or remember when you should take the next dose. All documents cited in this document are hereby incorporated by reference. The examples presented below are intended to illustrate particular embodiments of the invention, and are not intended to limit the scope of the specification, including the claims, in any way. Some abbreviations used in this application are described below: SM, sphingomyelin; SPT, serine palmitoyl transferase; LCAT, lecithin: cholesterol acyl transferase; LPL, lipoprotein lipase; PC, plasma phosphatidylcholine; RT-PCR, real-time polymerase chain reaction ApoE, Apolipoprotein E; WD, ApoE knockout mice fed diet feed Western WD + myr, ApoE knockout mice fed Western diet plus mlriocin; Normal, KO mice for ApoE fed normal or conventional feed; C57BI / 6J, wild type control mice fed normal or conventional feed; KO, knockout; TG, triacylglycerol; SRE regulatory elements of esteral; SREBP, protein binding element regulator of esterales. STD, conventional feed; LC / MS, Liquid Chromatography / Mass Spectroscopy Examples Materials - Cholesterol R1, triglyceride reagent and bovine serum albumin (BSA, without ultra-free fatty acids) were purchased from Roche Diagnostics Corporation (Indianapolis, IN). The chromatography column Superase 6HR was purchased from Pharmacia Biotech (Buckinghamshire, England). Sphinganine, sphingomyelin (brain), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine, and ceramide were purchased from Avanti Polar Lipids (Alabaster, AL). Myriocin, 1,2-hexadecanoiol, psychosine, serine, palmitoyl CoA, and OH Red O were obtained from Sigma (St. Lous, MO). The IHC-Cinc-Tris fixative was purchased from PharMingen (San Diego, CA). The normal feed and Western diet feed for rodents were obtained from Research Diet (New Brunswick, NJ). Water, acetonitrile and butyl alcohol (normal) quality for HPLC were from Mallinkrodt (Paris, Kentucky). The formic acid (90%) was from Aldrich (Milwaukee, Wisconsin). Ammonium acetate (P.M. 7-09), trimethylpentane, tetrahydrofuran, acetone, diclo-methane, and 2-propanol were obtained from EM Science (Gibbstown, New Jersey). The amyloid A serum ELISA kit for mice was obtained from Biosource (Camarillo, CA). Animal experiments - C57BI / 6J and KO male mice for ApoE in C57BI / 6J environment were obtained from Jackson Laboratoty (Bar Harbor, ME) or Taconic (Germantown, NY) (plaque formation in KO mice for ApoE is lipid-directed ( AS Plump et al., Cell., 1992; 71 (2): 343-53)). The myriocin was mixed with the Western diet containing 0.21% cholesterol and 21% fats. Mice 8-12 weeks old received 0.3 mg myocin / kg / day for 12 weeks (Table 1). KO mice for ApoE of 10-12 weeks of age (n = 8) were pre-fed on a Western diet for 2 weeks and the mice received various concentrations of myriocin for 4 weeks mixed with the diet. The control groups consisted of KO mice for ApoE fed normal diet or Western diet without mycocin, and C57BI / 6J mice fed normal diet. Each week body weight and feed were measured to examine feed intake. For the cuffed femoral artery model, KO mice were anaesthetized for male ApoE 8-10 weeks of age and the right femoral artery was dissected from its environment. A non-strangulating polyethylene sleeve (portex, 0.40 mm internal diameter, 0.80 mm external diameter, and 1.5 mm long) was placed, slightly tight around the right femoral artery. Table 1. Experimental design Group 1 2 3 4 Ñ 16 16 16 16 Cepa ApoE KO ApoE KO ApoE KO C57BI / 6J Conventional Western Western Conventional Diet Miriocin 0.3 mg / kg - Expression and activity of SPT - total RNA was isolated from the liver and aorta using Trizol (Invitrogen CA). The mRNA levels of LCB1 and LCB2 were measured by real-time polymerase chain reaction (PCR) in an ABI Prism 7900HT sequence detection system (Applied Biosystems, Foster City, CA). The following primers and series of probes were used: LCB1, forward primer 5'-CCGCTCCTTCGTGGTTGA-3 ': reverse primer, 5'-GAGGTAACGAAGCAGAAAAGCAG-3': probe, 5'FAM-TCAGCGGCTCTCCGGTCAAGGAT-3 '; LCB2, forward primer, 5'-CTGGATGAGGCTCACAGCATT-3 ', reverse primer, 5'- CCTCAGGATCCAGGCCAA-3', probe, 5'FAM- CCTTCAGGGCGAGGCGTGGTAGAT-3 '. The optimal number of cycles was established for each gene product with uniform amplification. Each level of mRNA was expressed in proportion to RND ribosomal 18s or as a ratio to GAPDH RNA. Liver tissues from each group were homogenized and the activity of SPT was measured using serine 14C and palmltoyl CoA as substrates and a thin layer chromatography (TLC) analysis (K. Gable et al., J. Biol. Chem. 2000; 275 (11): 7597-603). Analysis of sphingolipids and phospholipids by LC / MS and HPLC - Total lipids were extracted by the modified Bligh-Dyer extraction procedure (EG Bligh and W J. Dyer, Can. J. Med. Sci. 1959; 37: 911-917 and DK Perry, A. Bielawska and YA Hannun, Methods Enzymol., 2000; 312: 22-31). A Quattro Ultima quadrupole mass spectrometer from Micromass (Waters Corp., Mllford, Massachusetts) was used with a conventional Z-spray ™ ion source, set to an electrospray positive ionization mode, with the MassLynx ™ software running version 3.5, for quantitative determinations. The source conditions were typically the following: 3.5 kV capillary, 110 ° C source temperature, 325 ° C desolvation temperature. The multipliers are adjusted to 650 V and the residence time for each analyte was 100 milliseconds. Ionic transitions from precursor to product were established through direct infusion of each compound in the mass spectrometer. The following ionic transitions were used for quantification. Sphingomyelin (704 -> 184 m / z), sphinganine (302 -> 284 m / z), ceramide (566 -> 264 m / z) and psychosine (462 -> 282 m / z) as a standard internal. For the instrument, at a cellular shock pressure of 2 x 10"3 mbar (2 x 10" 4 kPa) of argon, the cone and collision voltages were as follows: Sphingomyelin (45V, 25eV), sphinganine (45V, 15eV), ceramide (45V, 25eV) and psychosine (45V, 25eV). The liquid chromatography system consisted of Shimadzu twin pumps (Columbia, Maryland) of HPLC LC-10ADvp with an SCL-10Avp controller (flow rate 0.2 ml / minute), and a CTC PAL automatic sampler from LEAP Technologies (Carrboro, North Carolina). For the quantitative procedure, the column of analysis was a Phenomenex (Torrance, California) Polar-RP column (2.0 x 150 mm, 4 μm) with a MetaGuard Polaris C8 2.0 mm safety column from MetaChem (Torrance, California ) directly connected (5 μm). Mobile phase A consisted of water / acetonitrile / formic acid (60/40/0, 1) and mobile phase B was propanol. The HPLC pumps were programmed with a gradient for each injection to supply 98% mobile phase A (0-1 minute), mobile phase A a! 30% (1-2 minutes), mobile phase A at 30% (2-4 minutes), and mobile phase A at 98% (4-4.5 minutes). A sample volume of 2 μl was injected into the LC / MS / MS system. The final chromatographic retention times for sphingomyelin, sphinganine, psychosine (internal standard) and ceramide were 4.84 minutes, 5.43 minutes, 4.92 minutes, and 5.31 minutes, respectively. The lipid extracts were analyzed by HPLC and evaporative light scattering detector to determine the levels of sphingomyelin and phosphatidylcollna (PC) in plasma (R. Homan and MK Anderson, J. Chromatogr. B. Biomed, Sci. Appl. 1998; 708: 21-6). Measurement of serum lipids and amyloid A in plasma - mice were sacrificed by inhalation of CO2 and blood was collected through cardiac puncture. Total cholesterol and triglyceride concentrations in plasma were determined enzymatically in a Cobas Mira Plus autoanalyzer using Cholesterol R1 and Triglyceride Reagents, respectively (Roche Diagnostics, Indiana, USA). The colorimetric changes were measured at 500 nm. The lipoproteins were separated from the mouse plasma by rapid protein liquid chromatography using a Superase 6HR column. The distribution of cholesterol between lipoproteins was determined by an online post-column analysis (K.A. Kieft, T.M. Bocan and B.R. Krause, J. Lipid Res. 1991; 32: 859-66). Serum Amyloid A (SAA) protein in plasma was measured by ELISA according to the manufacturer's instructions (Biosource). Vascular pathology - for a quantitative analysis of the diffusion of the atherosclerotic lesion, the mice sacrificed with saline solution were perfused and the aorta was isolated from the heart to the iliac bifurcation by sectioning small extensions of the artery and dissecting the tunica adventitia. After 24 hours of fixation with 10% buffered formalin, the aorta was opened longitudinally and fastened on the black wax. The lipids were stained with Oil Red O and photographs were taken. The percentage of aorta stained with Oil Red O was determined by Image Pro Plus image analysis software. For histological analysis, the mice were perfused and fixed in Cinc-Tris fixative, the sections embedded in paraffin were stained with Masson's Trichrome. The intimal layer macrophages were stained immunohistochemically using MAC-2 antibody (M3 / 38 clones from Cedarlane Laboratories Limited), counterstained with Verhoeff's elastic stain. The T lymphocytes were stained immunohistochemically with rat CD3 antibody (clones CD3-12, Serotec). The thickness of the lesion and the area occupied by the macrophages were determined using Image Pro Plus software. Statistics - The results are expressed in the form of the mean ± ETM. The statistical significance of the differences between the values of the mean was analyzed using the matched T-test. Comparisons between several groups were determined by one-way ANOVA with Dunnet's post hoc analysis using PRISM 2.01. If there was a significant difference between the groups, multiple comparisons of free distribution were made to find significance between the groups. When the typical errors of the mean were not equal, a nonparametric test (Mann-Whitney) was used to calculate the level of significance. The results were considered significant at P <; 0.05. The following Procedures were used in the indicated figures. Figure 1. Sphingomyelin Biosynthetic Route. Serine palmltoyl transferase (SPT) is the first step limiting the speed of synthesis of sphingolipids. Myroiocin specifically inhibits the SPT reaction. Figures 1-A, 1-B, 1-C, 1-D and 1-E. Gene expression and enzymatic activity of SPT. The mice were treated with 0.3 mg / mg / day of myriocin for 12 weeks by mixing it with Western diet feed. The liver was isolated, and total and homogenate mRNA were prepared without cells. The expression of SPT mRNA was quantified by quantitative RT-PCR. The expression was described as a ratio of mRNA of LCB1 (A) or LCB2 (B) to 18s RNA or to GAPDH RNA of rodents (n = 5, P> 0.05). The SPT activity of the homogenate without cells (C) was measured with 14C-labeled serine and palmitoyl-coenzyme A as substrates, and analyzed by TLC. The relative amounts of 3-ketosphinganine were determined by densitometry examination. The values indicated were the mean ± SEM (n = 3, * P <0.05). Figure 2. Distribution of lipoproteins in plasma of KO mice for ApoE fed Western diet. After 4 weeks of treatment with myroxine mixed with the diet, the mice were sacrificed and the plasma was isolated for the lipoprotein composition. The ßVLDL (A), LDL (B) and HDL (C) lipoproteins were separated from the mouse plasma by fast protein liquid chromatography (FPLC) using a Superase 6HR column. The distribution of cholesterol among the lipoproteins was determined by an in-line post-column analysis (Kieft, K.A., T.M.A. Bocan, B.R. Krause, J. Lipid Res. 1991.32: 859-866). The indicated values are the mean ± ETM (n = 8, * P < 0.01). Figure 3. Cholesterol and triglycerides in plasma of KO mice for ApoE fed a Western diet. After 4 weeks of treatment with myroxine mixed with the diet, the mice were sacrificed and the plasma isolated. The total cholesterol (A) and triglyceride (B) concentrations in plasma were determined enzymatically in a Cobas Mira Plus autoanalyzer using procedures of Cholesterol R1 and Triglyceride Reagents, respectively. The colorimetric changes were measured at 500 nm. The indicated values are the mean ± ETM (n = 8, * P < 0.01). Figure 4. Sphingomyelin in plasma and liver in KO mice for ApoE fed a Western diet. After 4 weeks of treatment with myroxine mixed with the diet, the mice were sacrificed and the plasma and liver were isolated. The total lipids were extracted by Chloroform: Methanol: Water (1: 1: 0.9) followed by phase separation. Sphingomyelin levels in plasma (A) and liver (B) were determined by LC / MS. The indicated values were the mean ± SEM (n = 5, p <0.05). Figure 5. Development of lesion in the femoral artery with cuff of KO mice for ApoE Fed with a Western Diet. The mice were anesthetized and the right femoral artery was dissected from its surroundings. A polyethylene cuff (Portex, internal diameter 0.40 mm, external diameter 0.80 mm, and length 1.5 mm) was placed lightly around the right femoral artery. The mice with KO cuff for ApoE were fed a Western diet mixed with myriocin at various concentrations for 4 weeks. The mice were sacrificed and the femoral artery was isolated and embedded in paraffin. The cross sections of the femoral artery were stained with Masson's Trichrome or Mac II antibody. The atherosclerotic lesion (black bar) and the size of macrophages (gray bar) in the femoral artery were quantified using Image Pro Plus software (Figure 5A). Serum amyloid A levels in plasma were determined by colorimetric ELISA (Figure 5B). The values indicated were the means ± ETM (n = 6-8, * P <0.05). The bar represents 100 μm. Figure 6. Distribution of Lipoproteins in Plasma of KO Mice for ApoE Fed with a Western Diet. After 12 weeks of treatment with myroxine mixed with the diet (0, 3 mg / kg / day), the mice were sacrificed and the plasma isolated for the composition of llpoproteins. The ßVLDL (A), LDL (B) and HDL (C) lipoproteins were separated from the mouse plasma by fast protein liquid chromatography (FPLC) using a Superase 6HR column. The distribution of cholesterol between the lipoproteins was determined by an online post-column analysis (Kieft, K.A., T.M.A.

Bocan, B.R. Krause, J. Lipid Res. 1991, 32: 859-866.). The reported values are the mean ± SEM (n = 5, * P <0.01, Western diet versus other study groups, n = 5, #P <0.05, Western diet plus myrocine versus normal feed ). Figure 7. Concentration of Cholesterol and Triglycerides in plasma. After 12 weeks of myocycin treatment, the mice were sacrificed and the plasma isolated. The plasma concentrations of cholesterol (A) and total triglycerides (B) were determined enzymatically in a Cobas Mira Plus autoanalyzer using R1 Cholesterol and Triglyceride Reagents procedures, respectively. The colorimetric changes were measured at 500 nm. The indicated values are the mean ± ETM (n = 5, * P < 0.01). Figure 8. Concentrations of Sphingomyelin in liver, plasma and aorta. After 12 weeks of treatment with myroxine, the mice were sacrificed and plasma, liver and aorta were isolated. The total lipids were extracted by a modified Blier-Dyer procedure. Sphingomyelin levels in liver (A), plasma (B), and aorta (C) were determined by LC / MS. The Indicated values are the mean ± ETM (n = 5, * P <0, Q5). Figure 9. Concentrations of sphinganine in the liver and aorta. After 12 weeks of treatment with myroxine, the mice were sacrificed and the plasma, liver and aorta were isolated. The total lipids were extracted by the Blier-Dyer procedure, the sphinganine levels in liver (A) and in aorta (B) were determined by LC / MS. The indicated values are the mean ± ETM (n = 5, * P < 0.05). Figure 10. Deposition of Lipids in AOrtas of KO mice for ApoE fed with Western Diet. KO mice were fed ApoE with Western diet in the presence or absence of myocin for 12 weeks. The mice were sacrificed and fixed with 10% buffered formalin for 24 hours. The aorta was dissected from the heart to the iliac bifurcation, opened along the ventral surface and held in a bottom of black wax. The accumulated lipids were visualized by staining with Oil Red O. The areas of the atherosclerotic lesion were quantified using Image Pro Plus and were represented as a percentage of the lesion area versus the total area of the aorta. The values indicated are the mean ± ETM (n = 4, WD vs. WD + myrvocine or normal feed, * P <0.01, Normal vs. WD + myriocin, #P < 0.01). The bar represents 1 cm. Figures 11 and 12. Formation of atherosclerotic lesions in the root of the aorta and in the brachiocephalic artery. KO mice for ApoE fed Western diet in the absence or presence of myriocin, KO mice for ApoE and C57BI / 6J mice fed normal feed were sacrificed and fixed with Zinc-Tris. The cross section of the brachiocephalic artery was stained by Masson's Trichrome and MAC-2 antibody and counterstained with Verhoeff's elastic stain. The atherosclerotic lesion (black bars) and the size of the macrophage (gray bars) in the brachiocephalic artery and at the root of the aorta were quantified using Image Pro Plus. The values indicated were the mean ± SEM (n = 5, * P <0.01, WD versus WD plus myxocin, #P <0.01, normal versus WD plus myriocin). The bar represents 100 μm. Figure 13. Proportion of SM / PC and ceramide concentrations in Plasma. The concentrations of SM and PC in plasma were determined, and the SM / PC ratio (Figure 13A) was calculated using HPLC. Plasma ceramide levels (Figure 13B) were analyzed by LC / MS / MS. The values are the mean ± SEM (n = 5; * P <0.01, Western diet versus Western diet plus myxocin, #P <0.05, conventional diet versus Western diet plus myrocin). Figure 14. Incorporation of T lymphocytes in the lesion of the aortic root. The cross section of the aortic root was stained with rat CD3 antibody and developed by diaminobenzidine (brown color) to detect incorporated T lymphocytes. The sections were counterstained with Harris hematoxylin (blue). The incorporation of T lymphocytes was quantified by measuring the number of T lymphocytes of the intimal layer in the root of the aorta (Figure 14). The values are the mean ± ETM (n = 5; * P < 0.05, Western diet versus normal feed; #P < 0.05, Western diet plus myriocin versus conventional diet). The bar represents 50 μm. The following results were obtained as indicated in the aforementioned Figures: Gene Expression and SPT Enzymatic Activity - RT-PCR analysis showed that myriocin had no effect on mRNA expression of LCB1 and LCB2 (Fig. 1A, B , D and E) in the liver. Compared to C57BI / 6J mice, SPT activity was increased in KO mice for ApoE fed a Western diet and normal feed for 12 weeks by 60% (n = 3, P <0.05) and 43% ( n = 3, p <0.05), respectively (Fig. 1C). Myriocin drastically reduced the activity of SPT in the liver of KO mice for ApoE fed Western diet (66% decrease compared to the KO mouse for ApoE fed Western diet without treatment and 48% compared to control group C57BI / 6J (n = 3, P <0.05)). Therefore, treatment with myocin had no effect on the expression of SPT, but was extremely effective in decreasing the enzymatic activity of SPT in the liver. Lipid composition. Treatment with mycocin significantly lowered plasma cholesterol and triglyceride levels in a dose-dependent manner (Figure 3). Plasma cholesterol levels were significantly affected by the inhibition of sphingolipid biosynthesis. At 0.1 mg myocyte / kg / day, the plasma cholesterol level was reduced by 46% compared to the control without myocin and reached a maximum of 76% decrease at a dose of 0.3 mg mycorino / kg / day (Figure 3A). In comparison with cholesterol, the degree of the reducing effect of TG by myriocin was smaller. Although there was no effect of myriocin on plasma TG levels at 0.1 mg / kg / day, TG levels in plasma were reduced by 44% at 0.3 mg / kg / day (Figure 3B). In addition, myriocin drastically reduced VLDL and LDL cholesterol by 83% and 63%, respectively (Figure 2A, B). In contrast, HDL cholesterol was increased 2.1 fold by Inhibiting SM synthesis (Figure 2C). Therefore, mycocin, an SPT inhibitor, significantly influenced the lipid profile in plasma.

To examine the effect of mlriocin on sphingolipid biosynthesis, sphingomyelin (SM) levels in plasma and liver were measured by LC / MS. In plasma, a treatment with myroxine at 1 mg / kg / day lowered sphingomyelin levels by 70% (Figure 4A). In contrast, sphingomyelin levels in the liver were decreased to a maximum of 46% with myroxine at 0.1 mg / kg / day. In the liver at 1 mg / kg / day, SM levels were comparable to those in the untreated group (Fig. 4B). Therefore, the inhibition of SPT during 4 weeks lowered the general lipid levels in plasma and liver in a dose-dependent manner. To investigate whether myrocyne was effective in reducing sphingomyelin biosynthesis, various concentrations of mycocin were administered to KO mice for ApoE for 4 weeks, and plasma SM levels were examined. HPLC analysis of plasma lipids showed that myriocin drastically reduced plasma SM levels in a dose-dependent manner (Table 2). At the highest dose of myroxine (1 mg / kg / day), plasma sphingomyelin levels were reduced by 70% compared to the control without myocin. Carrier, the myriocin mixed with the diet was effective in inhibiting the biosynthesis of sphingomyelin and lowering plasma sphingomyelin levels. Since the levels of MS in relation to the phospholipids have been considered as a risk factor for coronary artery disease, plasma PC levels were determined by HPLC analysis. Although there were significant changes in plasma SM levels, plasma PC levels did not change to the same extent by myriocin. Accordingly, the molar ratio of SM / PC was lowered by myocin in a dose-dependent manner (Table 2). Therefore, myrhoxine exerted a profoundly lowering effect of MS without significantly affecting the biosynthesis or PC degradation. Table 2. Levels of sphingomyelin (SM), phostatidylcollna (PC) in plasma of KO mice for ApoE.

WD, KO mice for ApoE fed Western diet; myr, myriocin. to myriocin was administered mixed with the diet for 4 weeks b P < 0.05, n = 10, versus a WD group Atherogenesis in the cuffed femoral artery - to determine the lipid-lowering effect of myriocin on atherogenesis, the femoral artery of KO mice was surrounded for ApoE using a non-polyethylene cuff Strangler In addition to the diet with high lipid content (Western diet) the strangulation with artery cuff accelerates the development of atherosclerosis. After 4 weeks of Western diet, the femoral artery with cuff of KO mice for ApoE developed lesions similar to atherosclerotic until an almost total occlusion of the lumen, mainly composed of macrophages (Fig. 5). In contrast, treatment with myroxine (0.1 mg / kg / day) reduced the development of atherosclerotic lesions and the accumulation of macrophages by 43% and 47% respectively (Figure 5A). At a myroxine dose of 0.3 mg / kg / day, the area of injury was reduced by more than 98% compared to the control without myocinin (Figure 5A). Plasma SAA levels that reflect the involvement of the inflammatory response were also measured. Treatment with myocycin reduced the SAA in plasma by 84% (Figure 5B). Therefore, myriocin reduces the atherogenesis of the femoral artery with cuff of KO mice for ApoE by a lipid lowering effect and the reduction of inflammatory protein levels. Cholesterol and triglycerides in plasma - To determine the effect of myxocin on lipoprotein metabolism, the lipoprotein cholesterol profile was examined using plasma isolated by FPLC (fast resolution liquid chromatography). Treatment with myriocin lowered ßVLDL and LDL cholesterol by 51% and 35%, respectively (Figure 6A, B), compared to KO mice for ApoE fed Western diet. By contrast, the content of HDL cholesterol was increased by 54% (Figure 6C). The distribution of cholesterol in the lipoproteins of KO mice for ApoE fed with conventional feed was comparable to that of the KO mice for ApoE treated with myriocin. Compared with the KO mice for ApoE, the WT C57BI / 6J wild type mice showed a very low total plasma cholesterol level. The majority of the cholesterol content in C57BI / 6J mice was in HDL (55.3 mg / dl in total plasma cholesterol, 58.4 mg / dl). In addition, myriocin lowered plasma apoB levels, which were comparable to those of the group fed conventional feed. Since levels of apoB in plasma, especially levels of apoB 100, in LDL correlate with atherogenesis (K. Skalen et al., Nature.; 417: 750-4), the reducing effect of apoB may contribute to the prevention of atherogenesis by myocin. Therefore, the inhibition of sphingolipid biosynthesis has a significant effect on the distribution of cholesterol in plasma lipoproteins. Since the SM content of lipoproteins affects the activities of the enzymes involved in lipid metabolism in vitro, it was questioned whether the inhibition of sphingolipid biosynthesis affected total cholesterol and triglyceride (TG) levels in plasma. Cholesterol (Fig. 7A) and TG (Fig. 7B) in plasma were the highest in KO mice for ApoE fed Western diet and the lowest in C57BI / 6J control mice with KO mice for ApoE fed with feed conventional between them. Myriocin showed significant lipid lowering activity by carrying both parameters to the levels of KO mice for ApoE fed conventional feed. Myriocin lowered cholesterol and TG in plasma by 41% and 45%, respectively (Figure 7). Therefore, it appears that myriocin lowered the overall lipid levels by affecting the activity of the enzymes involved in lipid metabolism. Biosynthesis of sphingolipids. Although the levels of SM are determined both by synthesis and by degradation, in the experimental system of the invention, the changes of SM were generally associated with changes in the activity of SPT and production of sphinganine, thus highlighting the role of the route Synthetic dependent SPT. Specifically, the levels of SM in the liver of C57BI / 6J mice were significantly lower than those of KO mice for ApoE fed Western diet (both treated with myriocin and fed conventional feed). Treatment with mycocin significantly lowered the accumulation of MS in liver compared to KO mice for ApoE fed Western diet (Figure 8A). In addition, KO mice for ApoE fed Western diet showed the highest levels of plasma SM, 33 times higher than C57BI / 6J mice and more than twice as high as KO mice for ApoE fed conventional feed (Figure 8B). Treatment with myocycin reduced plasma SM in KO mice for ApoE fed Western diet by 64% taking it to the level of its counterparts fed conventional feed. Small differences were observed between aortas of different treatments. However, there were statistically significant differences between KO mice for ApoE fed Western diet and C57BI / 6J control mice. The mlriocin decreased the SM levels by 20% (Figure 8C). Therefore, the inhibition of SPT by myriocin drastically affected the production of MS and the accumulation in the liver, plasma and aorta. Certain characteristics of SM can determine its fate and its potential role in atherosclerosis. It is known that secretory SMase causes hydrolysis of SM to generate ceramide, which stimulates lipoprotein aggregation and the formation of foam cells (S.L. Schissel et al., J. Biol. Chem. 1998; 273: 2738-46). A high proportion of SM / PC in lipoproteins determines their susceptibility to SMase. The ratio of SM / PC in plasma was measured using HPLC. Although, compared to the Western diet-fed group, plasma SM and PC levels were substantially lower in the myocynin-treated group, myocin did not affect the ratio of SM / PC in plasma (Figure 13A). On the other hand, treatment with myroxine reduced ceramide levels by 60%, which is comparable to the group fed conventional feed (Figure 13B). The lowest levels of ceramide were found in the control group C57BI / 6J. Therefore, the reduction of mycein-induced SM accumulation was accompanied by a substantial reduction in ceramide levels and was not associated with any change in the SM / PC ratio. Synthesis, accumulation and characteristics of the SM. To determine whether the inhibition of SPT activity resulted in the inhibition of SM production, the amount of sphinganine, an intermediate in SM synthesis (Figure 1) and a downstream marker of SPT activity, was measured. that can not be influenced by the degradation of SM by SMase. In the liver, sphinganine levels increased significantly in KO mice for ApoE fed Western diet as well as in KO mice for ApoE fed with conventional feed compared to C57BI / 6J control mice indicating an increase in the rate of SM synthesis in This model of atherosclerosis. Treatment with mycocin lowered sphinganine levels in the liver by 42% compared to KO mice for ApoE fed Western diet (Figure 9A). In the aorta, sphinganine levels in the KO mice for ApoE treated with myroxine, KO mice for ApoE fed with conventional feed, and control mice C57BI / 6J were reduced by 45%, 54% and 63%, respectively, in comparison with the group fed Western diet (Figure 9B). Sphinganine in plasma was below detectable levels. Therefore, treatment with mycocin inhibited the synthetic route of MS in both the liver and the aorta. Atherosclerosis Staining with Oil Red O on the surfaces of the aortas showed that the treatment with myriochne reduced the diffusion of the atherosclerotic lesion in KO mice for ApoE fed with the Western diet by 93%. If compared to KO mice for ApoE fed with normal feed, the atherosclerotic lesion of the KO mice for ApoE treated with myriocin was reduced by 75% (Figure 10). In addition, the growth of atherosclerotic lesions in the brachiocephalic artery and the area of the aortic valve were also significantly inhibited (Figures 11 and 12). At the root of the aorta, the area of injury was reduced by 44% and the area of macrophages decreased by 31% by treatment with myocin. In contrast, no change was observed between the KO mice for ApoE fed Western diet and the KO mice for ApoE fed normal feed (Figure 11). In the brachiocephalic artery, myocyte treatment leads to a 76% decrease in the area of injury and a 74% decrease in the area of macrophages (Figure 12). One point, the lesions in KO mice for ApoE fed with the Western diet, treated with myriocin, did not develop a necrotic nucleus. The myocynin treatment did not affect the accumulation of T cells (Figure 14). Therefore, the inhibition of SPT had substantial effects by lowering the lipids and anti-atherogenic. All publications, including but not limited to, issued patents, patent applications and journal articles, cited in this application, are incorporated by reference in their entirety herein. Although the invention has been described above with reference to the described embodiments, those skilled in the art will readily understand that specific detailed experiments are only illustrative of the invention.

Claims (15)

1. Use of a serine palmitoyl transferase inhibitor (SPT) for the preparation of a medicament for raising high density lipoprotein (HDL) particles in a mammal in need thereof.

2. Use of a serine palmitoyl transferase inhibitor (SPT) for the preparation of a medicament for lowering very low density lipoprotein (VLDL) particles and low density lipoprotein (LDL) particles in a mammal in need thereof.

3. Use of a serine palmitoyl transferase (SPT) inhibitor for the preparation of a medicament for lowering plasma triglyceride particles in a mammal in need thereof.

4. Use of a serine palmitoyl transferase inhibitor (SPT) for the preparation of a medicament for lowering serum total cholesterol levels in a mammal in need thereof.

5. Use of a serine palmitoyl transferase inhibitor (SPT) for the preparation of a medicament for improving the plasma lipid profile in a mammal in need thereof.

6. Use of a serine palmitoyl transferase (SPT) inhibitor for the preparation of a medicament for reducing the size of plaque in a mammal in need thereof.

7. Use of a serine palmitoyl transferase inhibitor (SPT) for the preparation of a medicament for reducing the size of an atherosclerotic lesion in a mammal in need thereof.

8. Use of a serine palmitoyl transferase (SPT) inhibitor for the preparation of a medicament for the treatment of a disease or condition selected from dyslipidemia, atherosclerosis, diabetes, metabolic syndrome or inflammation in a mammal in need thereof.

9. The use as in any one of the preceding claims, wherein the SPT inhibitor is myrocin.

10. A pharmaceutical composition comprising: a) a compound that is an inhibitor of serine palmitoyl transferase (SPT); and b) a second compound useful for the treatment of atherosclerosis or dyslipidemla.

11. A kit comprising: a) a serine palmitoyl transferase (SPT) inhibitor and a pharmaceutically acceptable excipient, carrier or diluent in a first unit dosage form; b) a second compound that is useful for the treatment of atherosclerosis or dyslipidemia and a pharmaceutically acceptable excipient, carrier or diluent in a second unit dosage form; and c) a means for containing the first and second unit dosage forms.

12. The composition of claim 10 or the kit of claim 11, wherein the second compound is an inhibitor of HMG-CoA reductase, an inhibitor of HMG-CoA synthase, or an inhibitor of the expression of the HMG-CoA reductase gene , an inhibitor of the expression of the HMG-CoA synthase gene, a CETP inhibitor, a bile acid sequestrant, an inhibitor of cholesterol absorption, an inhibitor of cholesterol biosynthesis, an inhibitor of squalene synthetase, a fibrate, niacin, a combination of niacin and lovastatin and an antioxidant; and a pharmaceutically acceptable excipient, vehicle or diluent in a second unit dosage form; wherein the amounts of first and second compounds result in a therapeutic effect.

13. The composition or kit of claim 12, wherein the second compound is an HMG-CoA reductase inhibitor selected from lovastatin, simvastatin, pravastatin, fluvastatin, atorvastatin, rivastatin, rosuvastatin or pitavastatin.

14. The composition or kit of claim 12, wherein the second compound is an inhibitor of CETP, which is torcetrapib.

15. The composition of claim 10 or the kit of claim 11, wherein the SPT inhibitor is myriocin.