Oxidised lipides and use thereof for treating inflammatory diseases and disorders

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to pharmaceutical compositions containing synthetic oxidised lipids.

EFFECT: developing the method for using the oxidised lipids for treating and preventing the inflammation associated with the endogenous oxidised lipid.

39 cl, 11 tbl, 15 ex, 25 dwg

 

The technical field to which the invention relates.

The present invention relates to new oxidized lipids and methods of use of oxidized lipids for the treatment or prophylaxis of inflammation associated with endogenous oxidized lipids. The methods of the present invention can be used to treat or prevent inflammatory diseases and disorders, such as, for example, atherosclerosis and related disorders, autoimmune diseases or disorders, and proliferative diseases or disorders.

Cardiovascular diseases represent a major risk factor for health in industrialized countries. Atherosclerosis, the most common cardiovascular disease is the main cause of heart attacks, strokes and gangrene limbs and, as a consequence, the main cause of death in the United States. Atherosclerosis is a complex disease caused by cells of many types and molecular factors (for more details see publication Ross, 1993, Nature 362:801-809). The process resulting from the destruction of endothelial cells and smooth muscle cells (SMC) of the arteries, includes the formation of deposits, consisting of fibrous, adipose and fibrous tissue, or plaques, preceding and accompanying inflammation. Destabilization of plaques can the t to cause further complications, such as rupture of blood vessels and venous thrombosis, resulting inflammatory and fibroproliferative response to many forms of damage. For example, it is assumed that the shear stress is the cause of frequent formation of atherosclerotic plaques in the blood circulatory system in the areas of occurrence of turbulent blood flow, such as branching vessels and structures of irregular shape.

First observed the phenomenon in the formation of atherosclerotic plaques is the adhesion of cells to areas of inflammation, such as macrophages generated from monocytes, endothelial layer of blood vessels and infiltration into the subendothelial space. Elevated levels of LDL in the plasma cause the deposition of lipids on the walls of blood vessels, resulting in adjacent endothelial cells produce oxidized low-density lipoprotein (LDL). In addition, the capture lipoprotein extracellular matrix leads to progressive oxidation of LDL lipoxygenase, active oxygen, peroxynitrite and/or myeloperoxidase. These oxidized LDL is then absorbed in large quantities by monocytes via phagocytic receptors expressed on their surfaces.

Filled with lipids monocytes and smooth muscle cells (SMC), referred to as kantamneni cells are the main component of fat is o strips. The interaction between Santanyi cells, endothelial cells and surrounding cells smooth muscle causes a state of chronic local inflammation, which ultimately leads to activation of endothelial cells, increased apoptosis of macrophages, proliferation and migration of smooth muscle cells and formation of fibrous plaques (Hajjar, D.P. and Haberland, M.E, J. Biol. Chem. 1997, Sep 12; 272(37):22975-78). As a result of destruction of plaque and blood clots is a blockage of the blood vessels, which prevents the blood flow and causes ischemia, a condition characterized by lack of oxygen in the tissues due to inadequate perfusion. When blocked the flow of blood to the heart through these arteries, the subject emerges “heart attack”; when there is occlusion of cerebral arteries, this subject arises stroke. When narrowing of the arteries that supply blood to the limbs, there is a strong pain, deteriorating physical mobility and may need amputation.

Oxidized LDL is involved in the pathogenesis of atherosclerosis and atherothrombosis, with effects on monocytes and smooth muscle cells, causing apoptosis of endothelial cells and disrupting the balance of anticoagulant in the endothelium. Oxidized LDL also inhibits anti-atherogenic HDL-associated destruction of oxidized phospholipids (Mertens, A and Holvoet, P. FASEB J. 2001 Oct; 15(12):2073-84). This Association is confirmed by many studies demonstrating the presence of oxidized LDL in the plaques in different animal models of atherogenesis and slow atherogenesis as a result of inhibition of oxidation of pharmacological and/or genetic means (see, for example, Witztum J. and Steinberg, D, Trends Cardiovasc Med 2001 Apr-May; 11(3-4):93-102 to familiarize yourself with the current scientific literature). Indeed oxidized LDL and malondialdehyde (MDA)-modified LDL has recently been proposed as an accurate marker of blood to determine the 1-St and 2-nd stage of coronary heart disease (U.S. patent No. 6309888 issued by Holvoet et al., and U.S. patent No. 6255070 issued by Witztum et al.).

Reduction of oxidation and the activity of LDL formed the basis of a number of proposed clinical methods of treatment and prevention of cardiovascular diseases. Bucala and others (Bucala et al., U.S. patent No. 5869534) described the ways modulation perechisleniya lipids by reducing the end product of progressive glycosylation, representing lipid characteristic of education xantana cells due to aging, disease, and diabetes. Tang and others (Tang et al., at Incyte Pharmaceuticals, Inc., U.S. patent No. 5945308) described a method for the identification and clinical application of the receptor for oxidized LDL person in the treatment of cardiovascular and autoimmune diseases, and t is the train of cancer.

Atherosclerosis and autoimmune diseases

Given the proposed role of a strong inflammatory fibroproliferative reaction in the case of atherosclerosis and ischemia, many scientists have attempted to determine the autoimmune component of vascular lesion. In the case of autoimmune diseases, the immune system recognizes and attacks the components that usually are not antigenic (autoantigens), in addition to the impact on penetrating into the body of foreign antigens. Autoimmune diseases are classified as a disease mediated by autoantibodies or cells. Typical autoimmune disease mediated by autoantibodies, are male and idiopathic thrombocytic purpura (ITP), and typical cladophoraceae diseases are Hashimoto thyroiditis and (juvenile) diabetes type I.

The recognition that immunopositive processes predominate in areas of atherosclerotic lesions, has made possible the study of lymphocytes and macrophages in the early stages, namely fat strips. It was found that these cells, which comprise the majority population of CD4+ cells (other cells are CD8+ cells)are more numerous than macrophages in the early stages of the lesions when compared with the later stages of defeat, when the ratio m is applied on the reverse. When studying the obtained results have questions about how does the phenomenon of primary immune sensitization potential antigen or an alternative is a simple epiphenomenon of previously induced local tissue damage. Regardless of the factors determining the content of the plaques at the initial stage of formation of these cells in areas of inflammation, they apparently characterize the activated condition defined by the concomitant expression of HLA-DR major histocompatibility complex class II receptor, interleukin (IL)and total leukocyte antigens (CD45R0) and integrin very late antigen-1 (VLA-1).

The inflammatory reaction that occurs in the early stages of atherosclerotic lesions may be primary triggering event, causing the production of different cytokines by local cells (i.e. endothelial cells, macrophages, smooth muscle cells and the cells of the zone of inflammation), or can be a form of protective reaction of the immune system on a dangerous process. Some cytokines, which, as has been established, more intensively are produced by resident cells include TNF-α, IL-1, IL-2, IL-6, IL-8, IFN-γ and chemoattractant of monocytes peptide-1 (MCP-1). In addition, it was established that the overexpression of platelet-derived growth factor (PDGF)and insulin-like growth factor (IGF) all cellular components in atherosclerotic plaques, that, apparently, increases prior inflammatory response in costimulated actions mitogenic and chemotactic factors. The Uemura and others (Uyemura et al., Cross-regulatory roles of IL-12 and IL-10 in atherosclerosis. J. Clin Invest. 1996, 97; 2130-2138) has established a pattern of T-cell type 1 cytokines in atherosclerotic lesions of humans, characterized by a strong expression of IFN-γ, but not mRNA of IL-4 compared with normal arteries. In addition, the affected area was detected overexpression of IL-12, growth factor T cells produced mainly by activated monocytes and selective inducer of the pattern of Th1 cytokines, as evidenced by the large number of its main heterodimeric forms of mRNA R and R40 (his dominant inducible protein).

Similarly, explicit evidence of the predominance of the cellular immune system in the atherosclerotic plaque, there are also numerous data confirming the participation of local humoral immune system. So, in the plaques was found deposition of immunoglobulins and the complement components in addition to increased expression of receptors for C3b and C3Bi in resident macrophages.

Valuable evidence of the involvement of immunopositive inflammation and atherosclerosis were obtained from studies of animal models. In mice with impaired immunological what aktivnosti (no MHC class I) atherosclerosis develops faster compared to immunocompetent mice. In addition, the introduction of the C57BL/6 mice (Emeson E.E., Shen M.L., Accelerated atherosclerosis in hyperlipidemic C57BL/6 mice treated with cyclosporin A. Am. J. Pathol 1993; 142:1906-1915) and new Zealand white rabbits (Roselaar SE, Schonfeld G., Daugherty A. Enhanced development of atherosclerosis in cholesterol fed rabbits by suppression of cell mediated immunity. J. Clin. Invest. 1995; 96:1389-1394) cyclosporine A, a potent suppressor of transcription of IL-2, caused significantly more severe atherosclerosis in the “normal” “load” lipoprotein. Recent research can shed light on the possible role of the immune system in combating spontaneous inflammatory process in the atherosclerotic plaque.

Atherosclerosis is not a classical autoimmune disease, although some of its manifestations, such as the formation of plaques, which seal the blood vessels, can be attributed to aberrant immunological reactivity. In classical autoimmune disease is very often possible to identify sensibilizarse autoantigen, attacked by the immune system and components of the immune system that recognize this autoantigen (humoral, i.e. autoantibodies, or cells, i.e. lymphocytes). In addition, it is known that this disease can be induced in healthy animals as a result of passive transfer of these components of the immune system, or in the case of humans, the disease can be transmitted from the pain is Oh pregnant woman to her child. Many of these features are absent in atherosclerosis. In addition, this disease is common risk factors such as hypertension, diabetes, reduced physical activity, Smoking, and other specified disease affects the elderly and is characterized by different genetic preference in comparison with classical autoimmune diseases.

The treatment of autoimmune inflammatory diseases may be directed to the suppression or modification of General and/or specific to a disease of the immune reactivity. For example, Aello (Aiello, U.S. patent No. 6034102 and 6114395) described the use of estrogen-like compounds for the treatment and prevention of atherosclerosis and the development of atherosclerotic lesions by inhibiting recruitment of cells to the area of inflammation. Similarly Medford and others (Medford et al., U.S. patent No. 5846959) described methods of preventing the formation of oxidized PUFA to ensure serdechno cardiovascular and other inflammatory diseases caused by intercellular adhesion molecule VCAM-1. In addition, Falb (Falb, U.S. patent No. 6156500) described a number of signaling molecules and molecules intercellular adhesion, are present in large numbers in the atherosclerotic plaque, as potential targets for anti-inflammatory therapy.

Since oxidized LDL certainly is involved in the pathogenesis is therosclerosis (see above), it was investigated the effect of the main components of the plaques on autoimmunity in health the development of atheromatous disease.

Immunological reactivity of the oxidized LDL. It is known that oxidized LDL (Ox LDL) exerts a chemotactic effect on T-cells and monocytes. It is also known that the Ox LDL and its by-products induce the expression of factors such as a chemotactic factor for monocytes 1, the secretion of colony stimulating factor and activating properties of platelets, which are potent stimulators of growth.

The active participation of immune response cells in atherosclerosis was justified by Stepmom S. and others (Stemme, S., et al., Proc. Natl. Acad. Sci. USA 1995; 92:3893-97)identified by the CD4+ clones plaques, reacting to Ox LDL as incentives. Clones corresponding to Ox LDL (4 of 27)were produced mainly γ-interferon, but not IL-4. Need to figure out whether this reaction by way of contacting the above-mentioned clones of T cells with immune system cells by stimulating immunogen (Ox LDL), or this reaction serves as a means of combating inactive atherosclerotic process.

Data related to the participation of humoral mechanisms and their significance, are much more controversial. In one recent study, it was reported elevated levels of antibodies about the Yves MDA-LDL, which is the metabolite oxidation of LDL in women suffering from heart disease and/or diabetes (Dotevall, et al., Clin. Sci. 2001 Nov; 101(5):523-31). Other researchers have demonstrated antibodies capable of recognizing many epitopes in oxidized LDL, indicating that the immune response and lipid components of apolipoprotein (Steinerova, A., et al., Physiol Res. 2001; 50(2): 131-41) in the case of atherosclerosis and other diseases, such as diabetes, renovascular syndrome, uremia, rheumatic fever and systemic lupus erythematosus. In several scientific publications have been noted elevated levels of antibodies to Ox LDL as atherosclerosis (defined by the degree of carotid artery stenosis, the severity of peripheral vascular disease etc). Recently, Scherer and others (Sherer et al., Cardiology 2001; 95(1):20-4) demonstrated elevated levels of antibodies to cardiolipin, beta 2GPI and OxLDL, with ischemic heart disease. Thus, there appears to be a consensus on the presence of antibodies to Ox LDL in the form of immune complexes in the atherosclerotic plaques, although the actual significance of this discovery is still not installed.

There is a hypothesis that antibodies to Ox LDL play an active role in lipoprotein metabolism. So, it is known that immune complexes of Ox LDL and corresponding antibodies more efficiently absorbed by macrophages in suspension compared to Ox LDL. From the top is open, you cannot draw any conclusions about the pathogenesis of atherosclerosis, because so far you have not answered the question whether the accelerated uptake of Ox LDL by macrophages favorable or harmful factor.

Important data concerning the importance of the humoral immune system in the process of atherogenesis, scientists are in the study of animal models. It is established that hyperimmunization rabbits with the absence of the LDL receptor homologous oxidized LDL induces the production of a large number of antibodies against Ox LDL, resulting in a significant decrease in the size of atherosclerotic lesions compared with the control group, which were injected with saline phosphate buffer (PBS). The reduction in plaque formation was also achieved by immunization of rabbits with liposomes with a high content of cholesterol while producing antibodies against cholesterol, and this effect was accompanied by a 35% decrease in cholesterol-containing lipoprotein very low density.

Thus, in laboratory and clinical studies have comprehensively demonstrated the pathogenic role of components of oxidized LDL and their value as autoantigens in the case of atherosclerosis and other diseases.

Mediated mucosal immunomodulation in the treatment of autoimmune diseases

In the recent past have been develo the Tana new methods and pharmaceutical preparations suitable for the treatment of autoimmune diseases (and related mediated T-cell inflammatory diseases, such as allograft rejection and neurological diseases caused by retroviruses). These treatments modulate the immune system, inducing tolerance to the disease, when administered orally or through the mucous membrane, for example, by inhalation, when used as tolerogenic proteins, antigens witnesses or suppress the disease of fragments or analogs of proteins or antigens witnesses. Such methods are described, for example, in U.S. patent No. 5935577 issued to Faneru and other Proteins and antigens-witnesses discussed below (for a General introduction to the tolerance induced through the mucous membrane, see publication Nagler-Anderson, C., Crit. Rev. Immunol. 2000; 20(2):103-20). It is established that intravenous administration of autoantigens (and their fragments containing the immunodominant epitope region of their molecules) causes suppression of the immune response under the action of the mechanism, referred to as clonal tolerance. Clonal tolerance deactivates only the T-cells of the immune attack that is specific to a particular antigen, resulting in a significant reduction in immune response to this antigen. Thus, T cells, stimuliruuschie autoim annou reaction and which is specific to autoantigen with induced tolerance, no longer proliferate under the influence of this antigen. This decrease in proliferation also reduces the immune response, determine the symptoms of autoimmune disease (such as damage to the nervous tissue, as seen in the case of multiple sclerosis). In addition, there is evidence that oral administration of autoantigens (or immunodominant fragments) as a single dose or in much larger quantities than those that run “active suppression”, can also induce tolerance due to anergy or deletion clone).

In scientific literature there is also described a method of treatment based on the active suppression. The mechanism of active suppression is different from the mechanism of clonal tolerance. The method is fully described in the patent application PCT/US93/01705, includes oral administration or introduction through the mucous membrane antigens that are specific to the tissue undergoing autoimmune effects. These antigens are referred to as “antigen-witnesses.” This treatment causes the induction of regulatory (suppressor) T cells in the lymphoid tissue associated with the gut (GALT), lymphoid tissue associated with the bronchi (BALT), or more often in the lymphoid tissue associated with the mucous membranes (MALT) (MALT includes GALT and BALT). These regulatory latkepalooza in the blood or lymphatic tissue and then migrate to the organ or tissue, amazed autoimmune disease, and inhibit autoimmune attack on the affected organ or tissue. T-cells detected by antigen-witness (who know at least one antigenic determinant of an antigen-witness used to identify them), directionally affect the hearth autoimmune attack, where they mediate local allocation of certain immune factors and cytokines, such as TGF-β is a growth factor (TGF-β), interleukin-4 (IL-4) and/or interleukin-10 (IL-10). Of the above factors TGF-β is an antigen-nonspecific immunosuppressive factor due to the fact that it suppresses the immune attack regardless of the antigen that triggers the attack (however, since tolerization antigen witness achieved by oral administration or by introduction through the mucous membrane, induces the release of TGF-β only in the immediate vicinity of autoimmune attack, it does not provide systemic immunosuppression). IL-4 and IL-10 are also antigen-non-specific immunoregulatory cytokines. IL-4, in particular, enhances the response of T cells-helper type 2 (Th2), i.e. the effect on the precursors of T cells and induces their differentiation with the formation preferably Th2-cells due to the reactions of Th1-cells. IL-4 also indirectly inhibits exacerbate the group caused Th1-cells. IL-10 is a direct inhibitor of the reactions Th1-cells. After oral tolerization mammal suffering from an autoimmune disease, using antigen-witnesses observed elevated levels of TGF-β, IL-4 and IL-10 in the hearth of the autoimmune attack (Chen, Y. et al., Science, 265:1237-1240, 1994). The mechanism of the suppression of antigen-witness confirmed von Garreton and others (von Herreth et al., J. Clin. Invest., 96:1324-1331, September 1996).

Oral immunomodulation, causing oral tolerance was effectively used in animal models for the treatment of inflammatory bowel disease by oral administration of probiotic bacteria (Dunne, C., et al., Antonie Van Leeuwenhoek 1999 Jul-Nov; 76(1-4):279-92), autoimmune glomerulonephritis by oral administration glomerular basal membrane (Reynolds, J. et al., J. Am. Soc. Nephrol. 2001 Jan; 12(1):61-70), experimental allergic encephalomyelitis (EAE, which is equivalent to multiple sclerosis or MS) by oral administration of the basic protein of myelin (MBP), adjuvant and collagen arthritis by oral administration to the subject accordingly collagen and HSP-65. In Boston Autoimmune company has conducted several experiments with participation of persons for the prevention of diabetes, multiple sclerosis, rheumatoid arthritis and uveitis. The results of experiments involving human subjects were less in akatsuki compared with experiments on animals, other than man, however, there has been some success in the prevention of arthritis.

Was also investigated immunomodulation by induction of oral tolerance to autoantigens found in the defeat of atherosclerotic plaques. The study of epitopes recognized by T-cells and Ig titers in clinical and experimental models of atherosclerosis identified three antigen candidate for suppression of inflammation in atheromatous lesions: oxidized LDL induced by stress itsokay protein HSP 65 and cardiodepressive beta-protein 2GP1. In the application for U.S. patent No. 09/806400 in the name of Shoenfeld and others (filed September 30, 1999), which is fully included in this description of the invention, the described reduction of approximately 30% of atherogenesis in the arteries in genetically predisposed transgenic mice with absence of LDL receptor-RD that oral was administered oxidized LDL person. However, this protective effect was achieved as the result of oral administration of the crude antigenic preparation comprising centrifuged, filtered and purified LDL from human serum, kotory were subjected to prolonged oxidation of Cu++or malondialdehyde (MDA). Although there was significant inhibition of atherogenesis due to oral tolerance, were not identificirebisatvis lipid antigens or immune and genetic components of LDL. Other of the obstacles were the instability inherent in the crude oxidized LDL in vivo due to the enzymatic activity, the uptake of oxidized LDL by the liver, immune mechanisms and cell heterogeneity, characterized by various donors. It is likely that sustainable, better defined similar oxidized LDL will ensure a more effective immunomodulation (e.g. due to oral tolerance).

Induction of immune tolerance and the subsequent prevention or inhibition of autoimmune inflammatory processes has been demonstrated with the introduction of suppressive antigens through the mucous membrane in addition to the mucous membrane of the intestine. The membrane around the eyes and the mucous membrane of the nasal cavity and intestines are exposed to many invasions, and exposure of autoantigens and have mechanisms of immunoreactivity. So, Rossi and others (Rossi, et al., Scand J. Immunol 1999 Aug; 50(2):177-82) found that the introduction of gliadin through the nose was as effective as intravenous administration in the weakening of the immune response to the antigen in the model diseases of abdominal cavity organs in mice. Similarly the introduction through the nose antigen receptor of acetylcholine was more effective than oral administration to slow down and reduce muscle weakness and proliferation of specific lymphocytes in mo who ate gravis in mice (Shi, F.D. et al., J. Immunol. 1999 May 15; 162(10):5757-63). Therefore, immunogenic compounds intended for insertion through the mucous membrane, for intravenous or intraperitoneal administration, must be optimally adapted to the ways of administration through the nose and other membranes.

Thus, there is an urgent need for new, well-defined, synthetic derivatives of oxidized phospholipids and related substances with higher metabolic stability and effective immunomodulation caused by, for example, oral, intravenous, intraperitoneal injection and injection through the mucous membrane.

Synthesis of oxidized phospholipids

Modified phospholipids have diverse applications. For example, the phospholipids that carry active compounds soluble in lipids, can be entered in compositions intended for percutaneous and transmembrane introduction (U.S. patent No. 5985292 issued Fournerou et al.), and derivatives of phospholipids can be introduced into liposomes and Biofactory intended for drug delivery (see, for example, U.S. patent No. 6261597 and 6017513, issued respectively to kurtze, Bebedero and others, and U.S. patent No. 4614796). In U.S. patent No. 5660855 described lipid structures of cholesterol, derived aminomalonate, which are enclosed in liposomes and fit the La directional influence on the cells or tissue of smooth muscle. These drugs reduce the restenosis of arteries methods RTSA. The use of liposomes for the treatment of atherosclerosis is further described in the application WO 95/23592 hope and Rodriguez, who suggested that the pharmaceutical compositions of the single-layer liposomes containing phospholipids. Liposomes, as described in the application WO 95/23592, you can optimize the flow of cholesterol from atherosclerotic plaques and are usually oxidized phospholipids.

It is known that derivatives of modified phospholipids, imitating the structure of the platelet activating factor (PAF), provide pharmaceutically active action in case of different disorders and diseases that affect functions such as vascular permeability, blood pressure, heart, etc. There is an assumption that one group of these derivatives may possess anticancer activity (U.S. patent No. 4778912 issued to Inoue et al.). In U.S. patent No. 4329302 described synthetic compounds phosphoglycerides, namely derivatives of lysolecithin, which mediate the activation of platelets. Although the compounds described in U.S. patent No. 4329302 are a simple 1-On-alkilany ether or acylphosphatase 1-O-fatty acids, found that the acylation short circuit lysolecithin allows to obtain compounds that activate platelets, in contrast to the acylation of a long chain, and that is rostau 1-On-alkilany ether biologically superior to acyl derivatives of 1-O-fatty acids from the point of view of simulation PAF.

The influence of patterns of different phospholipids on their biological activity was investigated Tokumura and others (Tokumura et al., Journal of Pharmacology and Experimental Therapeutics. July 1981, Vol. 219, No. 1) and described in U.S. patent No. 4827011 issued to Wisner and others, in relation to hypertension.

Another group of the ether derivatives of modified phospholipids described in Chinese patent No. 642665 issued to Berchtold. It was found that these ether derivatives of modified phospholipids suitable for use in chromatographic separation, but may have some physiological effect.

Oxidation of phospholipids occurs in vivo under the influence of free radicals and as a result of enzymatic reactions that often occur in the atherosclerotic plaque. The production of oxidized phospholipids in vitro usually includes a simple chemical oxidation of natural LDL or phospholipid component of LDL. Researchers studying the role of oxidized LDL, used, for example, iron ions and ascorbic acid (Itabe, H., et al., J. Biol. Chem. 1996; 271:33208-217)and copper sulfate (George, J. et al., Atherosclerosis. 1998; 138:147-152; Ameli, S. et al., Arteriosclerosis Thromb Vasc. Biol. 1996; 16:1074-79) to obtain molecules oxidized or moderately oxidized phospholipid, similar molecules associated with components of the plaques. It was found that similarly obtained identical molecules AU is oantigen, associated with atherogenesis (Watson A.D et al., J. Biol. Chem. 1997; 272:13597-607), and is able to induce a protective immune tolerance against atherogenesis (application for U.S. patent No. 09/806400 name Shoenfeld and others, filed September 30, 1999) in mice. Similarly Bunk (Koike engineering Germany, U.S. patent No. 5561052) describes a method of obtaining oxidized lipids and phospholipids using copper sulfate and peroxide-dismutase to obtain oxidized arachidonic or linoleic acid and oxidized LDL for diagnostic purposes. Davis and others (Davies et al., J. Biol. Chem. 2001, 276:16015) describe the use of oxidized phospholipids as agonists activated proliferation receptors in peroxisomes.

1-Palmitoyl-2-(5-oxovalerate)-sn-glycero-3-phosphocholine (POVPC, see example I for review with the description of two-dimensional patterns) and its derivatives, such as 1-Palmitoyl-2-glutaryl-sn-glycero-3-phosphocholine (PGPC), are typical examples of esterified oxidized phospholipids, which were investigated in connection with atherogenesis (see, for example, publications Boullier et al., J. Biol Chem. 2000, 275:9163; Subbanagounder et al., Circulation Research, 1999, pp. 311). It was also investigated the effect of other structural analogues related to this class of oxidized phospholipids (see, for example, Subbanagounder et al., Arterioscler. Thromb. Nasc. Biol. 2000, pp. 2248; Leitinger et al., Proc. Nat. Ac. Sci. 1999, 96:12010).

However, a disadvantage of the application in vio above oxidized phospholipids is the recognition, the binding and metabolism of the active ingredient in the body, resulting in importance dosing considerations and sustainability after injection.

In addition, the methods of oxidation are not sufficiently specific, resulting in the formation of different oxidation products, which makes necessary further purification or the use of untreated antigenic compounds. This problem becomes even more complicated in the case of natural LDL, even if he is cleared.

Thus, there is a recognized need for new synthetic oxidized phospholipids, improved methods of synthesis of these phospholipids and their use as immunomodulators in the absence of the above limitations.

The invention

One object of the present invention is a compound of General formula I

where

n is an integer from 1 to 6, and, if n=1, Cn, Bn, Rn, R n and Y are absent;

each of the B1B2, ...Bn-1and Bnindependently selected from the group consisting of oxygen, sulfur, nitrogen, phosphorus and silicon, each of the specified nitrogen, phosphorus and silicon substituted by at least one Deputy, selected from the group consisting of hydrogen, unshared pair of electrons, alkyl, halogen, cycloalkyl, the Rila, hydroxy, digitoxin, alkoxy, aryloxy, diarylike, dialkoxy and oxo;

each And1And2, ...An-1and Anindependently selected from the group consisting of CR"R"', C=O and C=S;

Y is chosen from the group consisting of hydrogen, alkyl, aryl, cycloalkyl, carboxy, sugar, phosphoric acid, phosphorylcholine, phosphorylethanolamine, phosphorylcholine, phosphorylethanolamine, fosfaurilirutisa, ethylphosphonic, phosphorylmethyl, phosphorylates, fosforilirovaniya, phosphorylmethyl, phosphorylethanolamine-N-lactose, phosphoethanolamine-N-[methoxy(propylene glycol)], postinst-4-phosphate, postinst-4,5-biphosphate, pyrophosphate, phosphatidylethanolamine, dinitrobenzenamine and fosfoglitserin; and

each of X1, X2,..., Xn-1independently is a saturated or unsaturated hydrocarbon of the General formula II

where

m is an integer from 1 to 26; and

Z is chosen from the group consisting of

where

W is chosen from the group consisting of oxygen, sulfur, nitrogen and phosphorus, each of the specified nitrogen and phosphorus substituted by at least one Deputy, selected from the group consisting of hydrogen, unshared pair of electrons, alkyl, halogen, cycloalkyl, aryl, hydroxy, digitoxin, alkoxy, aryloxy,diarylike, dialkoxy and oxo; and

at least one of X1, X2,..., Xn-1Z is not hydrogen;

and where

each of R1, R'1, R2, ...Rn-1, Rn, R'neach of R" and R"' and each of Ra, R'a, Rb, R b, ...Rm-1, R'm-1, Rmand R'mindependently selected from the group consisting of hydrogen, alkyl, alkenyl, quinil, cycloalkyl, aryl, heteroaryl, heteroalicyclic group, halogen, trihalomethyl, hydroxy, alkoxy, aryloxy, digitoxin, dialkoxy, diarylike, phosphonate, phosphate, phosphinyl, sulfonyl, sulfinil, sulfonamida, amide, carbonyl, thiocarbonyl, C-carboxy, O-carboxy, C-carbamate, N-carbamate, C-dicarboxy, S-dicarboxy and amino, or alternatively at least two of R1, R'1, R2, ...Rn-1, Rnand R'nand/or at least two of Ra, R'a, Rb, R b, ...Rm-1, R'm-1, Rmand R'mform at least one four-, five - or six-membered aromatic, heteroaromatic, alicyclic or heteroalicyclic ring; and

each of the1With2, ...Cn-1Cnand each of CA, Cb, ...Cm-1and Cmis chiral or organization of the achiral carbon atom, each chiral carbon atom has the S-configuration and/or R-configuration;

its pharmaceutically acceptable salt, prodrug, hitratio MES.

In accordance with other features of preferred embodiments of the invention, at least one And1And2... and An-1mean CR"R"', and at least one of A1And2... and An-1associated with X1, X2... or Xn-1that includes Z are not hydrogen.

In accordance with other features of preferred embodiments of the invention n is equal to 3 and at least one of A1and a2mean CR"R"'. Preferably And2mean CR"R"' and X2includes Z are not hydrogen. Further preferably both And1and a2mean CR"R"'.

In accordance with other features of preferred embodiments of the invention Z is selected from the group consisting of

where W means preferably oxygen and both R" and R"' are independently selected from the group consisting of hydrogen and alkyl.

In accordance with other features of preferred embodiments of the invention n is 1 and at least one of R1and R'1mean phosphate, or phosphonate.

In accordance with other features of preferred embodiments of the invention n is equal to 5 or 6 and at least one of R1, R'1and at least one of Rnand R'nform at least the one heteroalicyclic ring, for example, monosaccharide ring.

Another object of the present invention is a pharmaceutical composition containing as active ingredient the above-described compound and a pharmaceutically acceptable carrier.

In accordance with other features of preferred embodiments of the invention the pharmaceutical composition is packaged in a packaging material and identified in print, made in the packaging material or label printed on the specified packaging material, as a drug intended for the treatment or prophylaxis of inflammation associated with endogenous oxidized lipid, as described in detail below.

In accordance with other features of preferred embodiments of the invention the pharmaceutical composition further contains at least one additional compound suitable for the treatment or prophylaxis of inflammation associated with endogenous oxidized lipid, as described in detail below.

Another object of the present invention is a method of treatment or prophylaxis of inflammation associated with endogenous oxidized lipid, which includes an introduction to the needy to the subject a therapeutically effective amount of at least one oxidized lipid, which allows to treat or predator is to take the subject of inflammation, associated with endogenous oxidized lipid.

In accordance with other features of preferred embodiments of the invention, the oxidized lipid is chosen from the group consisting of oxidized phospholipid, platelet activating factor, plasmalogen, substituted or unsubstituted hydrocarbon containing 3-30 carbon atoms and the terminal of the oxidized groups, oxidized sphingolipid, oxidized glycolipid, oxidized membrane lipid and any analog or derivative.

In accordance with other features of preferred embodiments of the invention, the oxidized lipid is the above-mentioned General formula I.

In accordance with other features of preferred embodiments of the invention, the oxidized lipid is chosen from the group consisting of 1-Palmitoyl-2-azelaoyl-sn-glycero-3-phosphocholine, 1-hexadecyl-2-azelaoyl-sn-glycero-3-phosphocholine, 1-Palmitoyl-2-glutaryl-sn-glycero-3-phosphocholine (PGPC), 1-Palmitoyl-2-(5-oxovalerate)-sn-glycero-3-phosphocholine (POVPC), 1-Palmitoyl-2-(9-Oconnor)-sn-glycero-3-phosphocholine, 1-hexadecyl-2-acetyl-sn-glycero-3-phosphocholine, 1-octadecyl-2-acetyl-sn-glycero-3-phosphocholine, 1-hexadecyl-2-buterol-sn-glycero-3-phosphocholine, 1-octadecyl-2-buterol-sn-glycero-3-phosphocholine, 1-Palmitoyl-2-acetyl-sn-glycero-3-phosphocholine, 1-octadecenyl-2-acetyl-sn the gli is EPO-3-phosphocholine, 1-hexadecyl-2-(homogametic)-sn-glycero-3-phosphocholine, 1-hexadecyl-2-arachidonoyl-sn-glycero-3-phosphocholine, 1-hexadecyl-2-eicosapentaenoic-sn-glycero-3-phosphocholine, 1-hexadecyl-2-docosahexaenoyl-sn-glycero-3-phosphocholine, 1-octadecyl-2-methyl-sn-glycero-3-phosphocholine, 1-hexadecyl-2-butanoyl-sn-glycero-3-phosphocholine, Lyso PAF C16, Lyso PAF C18, 1-O-1'-(Z)-hexadecanoyl-2-[12-[(7-nitro-2-1,3-benzoxadiazole-4-yl)amino]-dodecanoyl]-sn-glycero-3-phosphocholine, 1-On-1'-(Z)-hexadecanoyl-2-oleoyl-sn-glycero-3-phosphocholine, 1-On-1'-(Z)-hexadecanoyl-2-arachidonoyl-sn-glycero-3-phosphocholine, 1-On-1'-(Z)-hexadecanoyl-2-docosahexaenoyl-sn-glycero-3-phosphocholine, 1-On-1'-(Z)-hexadecanoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine, 1-On-1'-(Z)-hexadecanoyl-2-arachidonoyl-sn-glycero-3-phosphoethanolamine and 1-On-1'-(Z)-hexadecanoyl-2-docosahexaenoyl-sn-glycero-3-phosphoethanolamine.

In accordance with other features of preferred embodiments of the invention the method further includes introducing to the subject a therapeutically effective amount of at least one additional compound intended for the treatment or prophylaxis of inflammation associated with endogenous oxidized LDL.

At least one additional compound is preferably chosen from the group comprising an inhibitor of HMGCoA-reductase (statins), mucosal adjuvant, corticosteron is d, the tool steroid anti-inflammatory therapy, a means of nonsteroidal anti-inflammatory therapy, analgetic, a growth factor, a toxin, HSP, beta-2-glycoprotein I, an inhibitor of protein, providing transfer complex cholesterolemia ether (CETP), agonist proliferating activated receptor peroxisome (PPAR), protivotromboznoe agent, an antiproliferative agent, ezetimib, nicotinic acid, an inhibitor of squalene, ApoE Milano and any derivative and similar.

Inflammation of the present invention associated with diseases and disorders such as, for example, idiopathic inflammatory diseases or disorders, chronic inflammatory diseases or disorders, acute inflammatory diseases or disorders, autoimmune diseases or disorders, infectious diseases or disorders, inflammatory malignant diseases or disorders, inflammatory diseases or disorders associated with transplantation, inflammatory degenerative diseases or disorders, diseases or disorders caused by hypersensitivity, inflammatory cardiovascular diseases or disorders, inflammatory cerebrovascular diseases or disorders, diseases or disorders peripheral vascular, inflammatory diseases or disorders jelly is, inflammatory gastrointestinal diseases or disorders, inflammatory skin diseases or disorders, inflammatory diseases or disorders of the liver, inflammatory neurological diseases or disorders, inflammatory diseases or disorders of the musculoskeletal system, inflammatory diseases or disorders of the kidney, inflammatory diseases or disorders of the reproductive system, inflammatory systemic diseases or disorders, inflammatory diseases or disorders of connective tissue, inflammatory tumors, necrosis, inflammatory diseases or disorders associated with the implant, inflammatory processes associated with aging, diseases or disorders caused by immune, proliferative diseases and disorders, and inflammatory lung disease or violations, as described in detail below.

The present invention allows to overcome the currently known drawbacks due to the creation of new synthetic oxidized lipids, to eliminate the limitations associated with presently known synthetic oxidized lipids and methods of treatment or prophylaxis of inflammation associated with endogenous oxidized lipid, synthetic oxidized lipids.

Except where specifically about uverennyh cases, all technical and scientific terms used in this description have the meanings commonly used by experts in the field to which this invention. The following describes acceptable methods and substances, although the practice or testing of the present invention it is possible to use methods and materials similar or equivalent described in this description of the invention. In the case of a conflict, you must contact the patent with the patent specification, including definitions of terms. In addition, substances, methods and examples are illustrative only and do not limit the scope of the invention.

Brief description of drawings

The present invention is described only in the form of an example with reference to the accompanying drawings. When detailed consideration of the drawings it should be emphasized that the individual parts are provided only as an example and to illustrate the preferred embodiments of the present invention and are presented with the most appropriate and easily understood description of the principles and conceptual objects of the present invention. In this regard it should be noted that the aim was not more detailed description of the structural details of the invention than is necessary for an informed understanding of the invention, with which ecialists in this area should be clear from the description of the invention in conjunction with the drawings, how to implement several variants of this invention.

Figure 1 shows the block diagram showing the synthesis of a simple ester of 2,5'-aldehyde-lecithin, 1-hexadecyl-2-(5'-oxapentane)-sn-glycero-3-phosphocholine (for D-ALLE) or 3-hexadecyl-2-(5'-oxapentane)-sn-glycero-1-phosphocholine (for L-ALLE) (ALLE) by the method of synthesis according to the present invention.

Figure 2 is a block diagram showing the synthesis of POVPC of the present invention.

Figure 3 shows a graph showing the inhibition of the initial stage of atherogenesis in mice with absence of apoE using intraperitoneal immunization mixed D - and L-isomers ALLE. Mice Apo-E KO at the age of 5-7 weeks were immunized with 150 μg/mouse mixed D - or L-isomers ALLE associated with purified derivative of tuberculin protein (ALLE L+D) (n=6), only cleaned derived protein of tuberculin (PPD) (n=5) were immunized or not (control group) (n=7). Atherogenesis is expressed as the area of atheromatous lesions in the aortic sinus in 4.5 weeks after the 4th immunization.

Figure 4 shows a graph showing the inhibition of the initial stage of atherogenesis in mice Apo-E KO oral introduction ALLE. Mice Apo-E KO at the age of 6-7,5 weeks oral introduced mixed D - and L-isomers ALLE: 10 μg/mouse (ALLE L+D 10 µg) (n=11) or 1 mg/mouse (ALLE L+D 1 mg) (n=11) or PBS (control group) (n=12) in a day h is for 5 days. Atherogenesis is expressed as the area of atheromatous lesions in the aortic sinus in 8 weeks after the last oral administration.

Figure 5 shows a graph showing the inhibition of the initial stage of atherogenesis in mice Apo-E KO oral and natalina the introduction of L-ALLE. Mice APO-E KO at the age of 7-10 weeks oral was administered 1 mg/mouse L-ALLE a day for 5 days (OT L-ALLE) (n=11) or were injected in the nose 10 μg/mouse L-ALLE a day for 3 days (NT L-ALLE) (n=11). Control mice orally was administered the same volume (0.2 ml) PBS (PBS ORAL) (n=12). Atherogenesis is expressed as the area of atheromatous lesions in the aortic sinus in 8 weeks after the last oral or nasal administration.

Figure 6 depicts a graph showing the suppression of immune reactivity against antigens of atherosclerotic plaques induced by oral administration of synthetic oxidized phospholipids L-ALLE and POVPC. Mice APO-E KO at the age of 6 weeks oral was administered 1 mg/mouse L-ALLE (L-ALLE) (n=2) or POVPC (POVPC) (n=3) in 0.2 ml PBS or only PBS (control group) (n=3) in a day for 5 days. One week after the last oral administration mice were immunized with a single subcutaneous injection of 50 μg of antigen oxidized LDL person. 7 days after received T cells from lymph nodes of inguinal region in accordance with the OPI is a W, see the following section “Materials and methods”, which was subjected to sensitizing antigen ox-LDL person for analysis of cell proliferation in vitro. Proliferation, which is an indicator of immune reactivity, expressed as the ratio of incorporation of labeled thymidine into the DNA of T-cells in the presence and absence of antigen ox-LDL person (stimulation index, S.I.).

7 depicts a graph showing the inhibition of the development of late stage of atherogenesis in mice APO-E KO by oral administration of synthetic oxidized phospholipids D-ALLE, L-ALLE or POVPC. Mice APO-E KO at the age of 24.5 weeks oral was administered 1 mg/mouse L-ALLE (L-ALLE) (n=11), D-ALLE (D-ALLE) (n=9) or POVPC (POVPC) (n=10) in a day for 5 days every 4 weeks for 12 weeks. Control mice orally was administered the same volume (0.2 ml) PBS (control group) (n=10) in accordance with the same scheme introduction. Atherogenesis is expressed as the area of atheromatous lesions in the aortic sinus 12 weeks after the first oral administration compared with estimates of lesions in non-immunized mice at the age of 24.5 weeks before the first oral administration (murdered in the 0-th time period).

On Fig depicts a graph showing the reduction in the content of triglycerides in VLDL in mice APO-E KO, induced by oral administration manufactured the definition of oxidized phospholipids D-ALLE, L-ALLE or POVPC. Mice APO-E KO at the age of 24.5 weeks oral was administered 1 mg/mouse L-ALLE (triangle) (n=11), D-ALLE (inverted triangle) (n=9) or POVPC (square) (n=10) in a day for 5 days every 4 weeks for 12 weeks. Control mice orally was administered the same volume (0.2 ml) PBS (circle) (n=10) in accordance with the same scheme introduction. The content of the triglyceride (Tg, mg/ml) was measured after 9 weeks after the time t=0 the enzymatic colorimetric method in the VLDL fractions after separation of the collected blood samples FPLC method in accordance with the description given in the following section “Materials and methods”.

Figure 9 depicts a graph showing the reduction of cholesterol in VLDL in mice APO-E KO, induced by oral administration of synthetic oxidized phospholipids D-ALLE, L-ALLE or POVPC. Mice APO-E KO at the age of 24.5 weeks oral was administered 1 mg/mouse L-ALLE (triangle) (n=11), D-ALLE (perevernutyi triangle) (n=9) or POVPC (square) (n=10) in a day for 5 days every 4 weeks for 12 weeks. Control mice orally was administered the same volume (0.2 ml) PBS (circle) (n=10) in accordance with the same scheme introduction. Cholesterol (cholesterol, mg/ml) was measured after 9 weeks after the time t=0 the enzymatic colorimetric method in the VLDL fractions after separation assembled who's blood samples FPLC method in accordance with the description, see the following section “Materials and methods”.

Figure 10 shows a two-dimensional structure of 1-hexadecyl-2-(5'-carboxybutyl)-sn-glycero-3-phosphocholine (CI-201, compound (VII), 1-hexadecyl-2-(5',5'-dimethoxyphenoxy)-sn-glycero-3-phosphocholine (compound VIIIa) and 1-hexadecyl-2-(5',5'-deoxypentose)-sn-glycero-3-phosphocholine (compound VIIIb).

Figure 11 depicts a graph showing the inhibition of the initial stage of atherogenesis in mice APO-E KO when oral administration of CI-201. Mice APO-E KO at the age of 12 weeks oral introduced CI-201: 0.025 mg/mouse (n=14) or 0.2 ml PBS (control group) (n=15) every day for 8 weeks (5 times a week). Atherosclerosis is expressed in the form of a square sarmatskogo lesions in the aortic sinus after 11 weeks after the first oral administration.

On Fig a-d presents photographs showing the expression levels of the cytokine in the aorta of mice that were injected ALLE, CI-201, it ETHYLACETYLENE derived (Et-acetal), it methylacetylene derived (IU-acetal), oxLDL or PBS. In particular, figures 12A and 12b shows an increase in the level of expression of IL-10 in the aorta of mice treated with ALLE, CI-201, Et-acetal, IU-acetal and oxLDL compared with control mice (PBS) and reduced levels of expression of IFN-gamma in aorta mice, which were injected ALLE, CI-201, BF-acetal and oxLDL compared with mice treated with PBS; the figures 12C and 12d dormancy is the result of a decrease in the expression of IL-12 in mice which was introduced ALLE, CI-201 and Et-acetal, compared with the group treated with PBS. Mice APO-E KO at the age of 10-12 weeks oral was administered 1 mg/mouse/0.2 ml of the test antigen (ALLE, CI-201, Et-acetal, IU-acetal) or 0.1 mg/mouse/0.2 ml oxLDL or injected 0.2 ml PBS. Oral administration produced 5 times a day, and the expression of cytokines was evaluated 8 weeks after the last oral administration.

On Fig depicts a bar graph showing the attenuation of atherogenesis in mice LDL-RD oral introduction oxLDL. Mice LDL-RD oral was administered PBS or 10, 100 and 1000 μg/dose oxLDL 5 times a day. Atherogenesis is expressed as the area of atheromatous lesions in the aortic sinus in 5 weeks after the last oral administration.

On Fig a-b depicts a bar graph showing the inhibition of atherogenesis in mice APO-E KO when oral administration of CI-201. Mice Aro-E TO oral was administered PBS (control group) or 0.1, 1 and 10 μg/dose of CI-201 in the form of three series at the beginning of each month, 5 times a day in each series. Atherogenesis is expressed as the area of atheromatous lesions in the aortic sinus 12 weeks after the first oral administration. Figure 14a shows the degree of atherosclerosis in each group. Figure 14b shows the strong influence of low doses of CI-201 on atherosclerosis compared to the “base” group (userswindows in the 0-th day) and to trolley group.

On Fig a-b depicts a bar graph showing increased levels of IL-10 in serum (figure 15A) and preventing the increase of SAA (figure 15b) in mice APO-E KO immunized with CI-201. Mice Aro-E TO oral was administered PBS (control group) or CI-201 5 times a day. Serum was collected at the beginning of the experiment, after 2 weeks and 4 weeks after the first oral administration. The levels of markers were evaluated in accordance with the description given in the following section “Materials and methods”.

On Fig a-b presents pictures (figure 16A) and chart (figure 16b), which shows the levels of expression of the cytokine in the aorta of mice treated with CI-201 or PBS. In particular, figures 16A and 16b shows an increased expression level of IL-10 in the aorta of mice treated with CI-201, compared to the control mice (PBS), and reduced levels of expression of IFN-gamma in aorta mice treated with CI-201, compared with mice that were administered PBS. Mice Aro-E TO oral was administered 1 mg/mouse CI-201 or 0.2 ml/mouse in PBS 5 times a day. The expression of anti-inflammatory cytokine IL-10 and Pro-inflammatory cytokine IFN-γ was determined in 8 weeks after the last oral administration.

On Fig presents photographs showing the results of oral administration of CI-201 targeted effect on the aorta, mice APO-E KO. Although the introduction of CI-201, affect and the mouth, caused increased expression level of IL-10 and decreased levels of expression of IFN-gamma compared with the introduction of the PBS, there were no differences in the expression of cytokines in the spleen and small intestine between the group treated with CI-201, and a control group treated with PBS.

On Fig shows a diagram of the study to assess the attenuation of adjuvant-induced arthritis (AIA) in rats which had previously introduced CI-201.

On Fig depicts a bar chart showing the results of oral administration of CI-201 rats suffering from adjuvant-induced arthritis (AIA), an estimate of the swelling of the paws. The Lewis rats orally introduced CI-201 or PBS (control group) 5 times a day, and then was intradermally injected with a suspension of tuberculosis microbacteria.

On Fig shows a diagram of the study to assess the attenuation of adjuvant-induced arthritis (AIA) in rats, which were constantly introduced CI-201.

On Fig depicts a bar chart showing the results of oral administration of CI-201 Lewis rats suffering from AIA-induced arthritis, an estimate of the swelling of the paws. The Lewis rats orally introduced CI-201 or PBS (control group) 5 times a day, after which they induced arthritis and constantly oral introduced CI-201 3 times a week.

On Fig shows comparative graphs showing the evaluation of arthritis, controllable is in the process of development of arthritis in rats which was introduced CI-201 in different concentrations, compared with rats that were administered PBS.

On Fig shows comparative graphs showing the percentage of rats with symptoms of arthritis after injection of PBS (control group) and CI-201 in different concentrations.

On Fig depicts a bar chart showing the effect on early stage of atherogenesis in mice APO-E KO, induced by oral administration of pre-oxidised compounds V. Female mice APO-E KO at the age of 8-10 weeks orally injected with a connection V: 5 mg/mouse (n=6), 1 mg/mouse (n=6), 0.2 mg/mouse (n=6) or PBS (control group) (n=7) in a day for 5 days. Atherogenesis is expressed as the area of atheromatous lesions in the aortic sinus in 8 weeks after the last oral administration.

On Fig depicts a bar chart showing the effect on atherogenesis in mice APO-E KO, induced by oral administration of pre-oxidised compounds V. Mice APO-E KO at the age of 23-26 weeks was slaughtered at the beginning of the experiment (base group B.L., n=10) or were injected them with PBS (control group, n=11) or 0.1 μg/dose of a compound V (n=10) in the form of three series at the beginning of each month 5 times a day in each series. Atherogenesis is expressed as the area of atheromatous lesions in the aortic sinus 12 weeks after the first oral in which edenia.

Description of the preferred embodiments of the invention

The present invention relates to compositions containing oxidized lipids and methods of use thereof for the treatment or prophylaxis of inflammation associated with endogenous oxidized lipids. In particular, the objects of the present invention are (i) new oxidized lipids; (ii) pharmaceutical compositions containing these lipids; (iii) the use of new oxidized lipids, and other oxidized lipids for the treatment or prophylaxis of inflammation associated with endogenous oxidized lipids, and, as a consequence, for the treatment or prevention of diseases and disorders characterized by inflammation, which include, but are not limited to, atherosclerosis, cardiovascular disease, cerebrovascular disease, peripheral vascular disease, stenosis, restenosis, stenosis in the stent, autoimmune diseases or disorders, inflammatory diseases or disorders, infectious diseases or disorders, and proliferative diseases or disorders.

Principles and methods of implementing the present invention can be better understood with reference to the accompanying drawings and the description.

Before proceeding to a detailed description of at least one variant of implementation and the attainment, it should be noted that this invention is not limited to application, discussed in detail in the following description or illustrated by examples. The invention includes other embodiments of or may be implemented in different ways. Furthermore, it should be understood that the phraseology and terminology used in the description of the invention, serves only to describe and does not restrict the scope of the invention.

Experimental and clinical data suggest direct involvement of oxidized LDL (ox LDL) and LDL components in the etiology of the increased inflammatory response in the case of atherosclerosis. Was demonstrated as the cellular and humoral immune reactivity against oxidized LDL, is associated with plaques, suggesting a significant autoimmune component against oxidized LDL in the process of atherogenesis. Thus, against LDL, oxidized LDL and their components were directed numerous methods of prevention and treatment of heart disease, cardiovascular disease and peripheral vascular disease.

In previously performed studies of the role of oxidized LDL and its components in the immune response to endogenous (e.g., associated with plaques) oxidized LDL was used crude antigenic preparation is aderrasi spin-on, filtered and purified LDL serum of a person who was subjected to a long process of oxidation of Cu++or MDA, or synthesized analogs of oxidized LDL. Because phospholipids are considered to be active components of LDL, studies using synthesized analogs of oxidized LDL typically include the use of oxidized phospholipids (for example, POVPC and PGPC).

Although previously it was thought that by oral administration of oxidized LDL can be achieved 30% reduction of atherogenesis, suggesting the protective effect of oxidized LDL, mainly due to oral tolerance, were not identified specific lipid antigens or immunogenic components of LDL. Another obstacle that had previously experienced researchers, is the volatility inherent in the crude oxidized LDL in vivo, due to the enzymatic activity and uptake of oxidized LDL by the liver and the immune mechanisms of the cells. Such volatility inherent in the use of in vivo synthetic derivatives of oxidized LDL, such as POVPC and PGPC (described above).

So still was not identified a direct correlation between exogenous oxidized LDL or its components and endogenous oxidized LDL in relation to immunomodulation. Not previously have also been found analogs of oxidized LDL, which are not p is Issa them instability and other restrictions, characteristic for the introduction of oxidized LDL, is able to modulate the immune and/or inflammation associated with endogenous oxidized LDL and other endogenous oxidized lipid.

In the creation process of the present invention, it has been assumed that the synthesized oxidized lipids in General and analogs of oxidized LDL, in particular, can modulate immune reactivity against endogenous oxidized lipids in General and oxidized LDL, in particular, and, thus, can be used for the treatment or prevention of many diseases and disorders characterized by inflammation and/or altered immune response, such as, for example, atherosclerosis and related diseases or disorders, and other diseases and disorders associated with endogenous oxidized lipid.

The inflammation that occurs in the process of atherogenesis, often leads to complications such as the destruction of platelets and thrombosis (Libby et al., Inflammation and atherosclerosis. Circulation 2002; 105:1135-1143).

The presence of activated T-lymphocytes at the site of atherosclerotic lesions person may indicate their involvement in the onset and progression of the disease (Ross R. Atherosclerosis-an inflammatory disease. NEJM. 1999; 340:115-126). The main class of T-lymphocytes, CD4+, can be differentiated along the lines of Th1 or Th2, which differ functionally produced what Fokina: interferon (IFN)-γ, secretively Th1 cells, and interleukin (IL)-4, secretively Th2 cells. The main inducers of Th1 cells and Th2, respectively), IL-12 and IL-10 (A. Daugherty and Rateri DL., T lymphocytes in atherosclerosis the Yin-Yang of Th1 and Th2 Influence on lesion formation, Circ. Res. 2002; 90:1039-1040; Hansson G.K., Vaccination against atherosclerosis science or fiction. Circulation. 2002; 106:1599-1601).

It is established that T-lymphocytes are isolated from whole blood of subjects suffering from acute coronary syndrome, or obtained from plaques of the carotid artery, specifically learn Ox LDL and proliferate under the influence of Ox LDL (Hansson G.K. Immune mechanisms in atherosclerosis. Arterioscler Thromb Vasc. Biol. 2001; 21:1876-90). Ox LDL and by-products of oxidized lipids (e.g., oxidized phospholipids) are present in atherosclerotic plaques (Witztum 2001, supra).

Therefore, despite the fact that oxidative modification of LDL may be a necessary condition for the rapid accumulation of LDL in macrophages and education xantana cells, it can also induce immunogenic epitopes in the molecule LDL, causing the formation of antibodies against Ox LDL.

Therefore, the epitopes of oxidized LDL serve as important ligands mediating the binding and excretion damaged by oxidation of lipoprotein particles and apoptotic cells and induce the natural immune response, causing their destruction. On the other hand, the epitopes of oxidized LDL may be involved in immune activation, which'hara is cherished growing atherosclerotic plaque.

Given the above, the authors of the present invention concluded that the compounds that can serve as epitopes of oxidized LDL, can modulate the immune response, providing a rather favourable than harmful effect on atherogenesis. In other words, it was found that introduction, preferably oral, analogs of oxidized LDL, such as oxidized phospholipids causes tolerance to endogenous oxidized LDL formed in the process of atherogenesis and, thus, reduces inflammation and slows the development of atherogenesis.

Recently published data confirming the possibility of using immunomodulation as a new therapeutic approach to the treatment of atherosclerosis (Nicoletti et al., Induction of neonatal tolerance to oxidized lipoprotein reduces atherosclerosis in Apo E knockout (Apo-E KO) mice. Mol. Med.2000; 6(4):283-290). It was found that intraperitoneal injection of oxidized LDL newborn mice (APO-E KO) caused T-cell tolerance due to deletion clone, weakening of the immune response to oxidized LDL and reduced susceptibility to atherosclerosis.

In the pathogenesis of atherosclerosis and many other diseases and disorders participates acquired and natural immunity. Taking into account the high content of lipids in the location of the injury in these lipids may be possible Misha is s for the immune response, caused by atherosclerosis. Recently, it was shown that the natural T-killer cells (NKT) can recognize lipid antigens presented by CD1 molecules. The CD1 molecules present lipid antigens to T cells in contrast to evolutionary related molecules of the major histocompatibility complex (MHC) class I and II, which show the peptide antigens. However, like the molecules of MHC class I molecules CD1 include a heavy chain associated with a light chain β2-microglobulin (β2M). Crystal structure of two isoforms CD1, human CD1b and CD1d mouse, characterized by a common organization of domains, resembling molecules MHC class I. In particular, antigennegative site in CD1 is hydrophobic, forming channels (CD1b) or pockets (mouse CD1d), which may be hydrocarbon chains of lipids. The narrow opening between the α helices allows you to display the polar part of the lipid in the area available for recognition receptors on T cells (TCR). This system facilitates the binding of different lipid molecules attached to different polar head groups, thus creating a large pool of potential CD1-presented antigens (Zeng et al., Crystal structure of mouse CD1: an MHC-like fold with a large considered are hydrophobic binding groove. Science 1997; 277:339-345).

The CD1 molecules bind antigens alien lipids as they examine endosome compartment is infected antigen presenting cells. Unlike T cells that recognize CD1-restricted alien lipids, CD1-restricted T cells that interact with proteins that function as “autorefraction”that can be quickly stimulated to perform helper and effector functions when interacting with CD1-expressing antigen-presenting cells. Functional differences between subpopulations CD1-restricted T cells and the ways in which these cells affect inflammatory and tolerogenic action of dendritic cells and activate natural killer cells and other lymphocytes, allow you to define how the CD1-restricted T cells regulate antimicrobial responses, antitumor immunity and the balance between tolerance and autoimmunity (Vincent et al., Understanding the function of CD1-restricted T cells. Nat. Immunol. 2003; 4:517-23).

Tupin and others (Tupin et al., CD1d-dependent Activation of NKT Cells Aggravates Atherosclerosis. J. Exp. Med. 2004; 199:417-22) investigated the role of CD1d-restricted natural T killer cells in atherosclerosis using mice with absence of the apolipoprotein E (apoE(-/-) mice and APOE(-/-), crossed with mice, CD1d(-/-), (CD1d(-/-)apoE(-/-)), which saw a 25% decrease in lesions compared with mice apoE(-/-). Introduction alpha-galactosylceramide, synthetic glycolipid, activating the natural T-killer cells via CD1d, caused a 50% increase is an increase in the size of lesions in mice apoE(-/-) and thus does not affect the size of lesions in mice apoE(-/-)CD1d(-/-). The results show that the activation of CD1d-restricted natural T killer cells aggravates atherosclerosis. Gow and others (Zhou et al., Editing of CD1d-bound lipid antigens by endosomal lipid transfer proteins. Science. 2004; 303:523-7) found that mice with no prosaposin, the precursor of a family of proteins that carry endosome lipids (LTP), are characterized by specific defects CD1d-mediated presentation of antigens and they have no natural T killer cells Vα14. In vitro saposin were extracted Monomeric lipids from membranes and CD1, thus stimulating the loading and editing of lipids in CD1. Temporary CD1 complexes, lipid and LTP involve the creation of a model “dragging”, in which the exchange of lipids between CD1 and LTP occurs on the basis of their respective affinity for lipids. LTP create a previously unknown link between lipid metabolism and immunity and, apparently, have a strong influence on the range of autoantigens, tumor antigens and antigens of microbial lipids.

Activated macrophages (M2) type 2 is an alternate path leading to the classical activation of macrophages. The M2 cells are antigen-presenting cells (APCS), which are present in a thin layer of the intestine in the form of a part associated with the intestinal immune system. Cells M2 react on the expression of IL-10 instead of the classic reactions of macrophages acetocina Th1, as is described below.

Activated macrophages are used as antigen-presenting cells (APC). Recognition of antigens by T-cells is the main event that control of the acquired immune response.

Classic by IFN-γ-dependent activation of the macrophage cells of the Th1-type is a well-known cell-mediated immunity. Activation of macrophages depends on products specifically activated T-cells, helper cells, Th1 lymphocytes and natural killer cells, in particular, IFN-γ and the network of cytokines, including IL-12 and IL-18 produced by ARS. In the last decade has been the recognition of the concept of alternative pathway activation of macrophages by cytokines of the Th2-type IL-4 and IL-13 with IL-10, which is considered defines a specific phenotype of macrophages, compatible with another role in humoral immunity and recovery.

IL-4 and IL-13 enhance the expression of mannose receptor and MHC molecules of class II macrophages that stimulate endocytosis and presentation of antigens.

Immunoglobulins and immune complexes can be contacted with the activating and any abscopal receptors for Fc and complement. In addition, ligation of Fc-receptor has a significant effect on the release of cytokines, such as IL-12/IL-10 and IL-4, antigen-presenting cells and other cells of the innate and acquired immune si theme (Gordon S. Alternative Activation of Macrophages, Nat. Rev. Immunol. 3:23-34; 2003).

Macrophages, activated causing inflammation stimuli (e.g., IFN-γ) and put in immune complexes, have quite the opposite effect instead of the response to Th1: at elevated levels of IL-12 and secondary levels of IL-10 is a significant decrease in levels of IL-12 and increased levels of IL-10. IL-10 exerts an immunosuppressive effect on macrophages (Anderson, C.F. and Mosser, D.M. A Novel Phenotype for an Activated Macrophages: the Type 2 Activated Macrophage J. Leukoc. Biol.72:101-106; 2002). IL-10 acts on the receptor plasma membrane, different from the receptors for IL-4 and IL-13, however different its influence on gene expression of macrophage, including a stronger inhibition of antigen-presenting and effector functions along with activation of selected genes or functions (Gordon S. Alternative Activation of Macrophages. Nat. Rev. Immunol. 3:23-34; 2003).

Therefore, we can conclude that in addition to the impact on atherosclerosis and other diseases that are directly associated with oxidized LDL, immunomodulation and anti-inflammatory effect caused by the (synthetic) analogs of oxidized LDL, can be used for the treatment and prevention of other diseases and disorders that are directly or indirectly associated with endogenous oxidized LDL and other oxidized lipids. This conclusion was confirmed by several studies is mi, the aim was to immunotherapy of autoimmune diseases such as rheumatoid arthritis (RA), diabetes type I and multiple sclerosis, by modulation of the individual immune pathways involved in inflammation, or acquisition of tolerance to different antigens (Bielekova et al. Encephalitogenic potential of the myelin basic protein peptide (amino acids 83-99) in multiple sclerosis: Results of a phase II clinical trial with an altered peptide ligand. Nat Med. 2000;6:1167-1175; Kappos et al. Induction of a non-encephalitogenic type 2 T helper-cell autoimmune response in multiple sclerosis after administration of an altered peptide ligand in a placebo-controlled, randomized phase II trial. Nat. Med. 2000;6:1176-1182).

Thus, there are numerous data that confirm the relationship between lipids, inflammation and the immune system and suggest a direct link between them.

Trying to improve the treatment of inflammations, diseases and disorders associated with oxidized lipids, the authors of the present invention have created a new synthetic oxidized phospholipids and structurally related compounds, have no limitations that are characteristic of oxidized LDL and other famous oxidized phospholipids and lipids (as identified above).

As shown in the “Examples”section, when implementing this izobreteniya in practice, it was confirmed that oral administration and/or introduction through the mucous membrane of new analogs of oxidized LDL module which induces immune and/or inflammatory response to endogenous oxidized LDL, resulting in the attenuation of inflammatory responses in inflammatory diseases such as atherosclerosis and rheumatoid arthritis. The obtained results clearly show the influence of exogenous oxidized lipids on inflammatory and immune processes involving endogenous oxidized lipid.

Thus, one object of the present invention are new compounds that mimic the immunomodulatory effects induced by oxidized LDL, and/or inflammation associated with oxidized LDL and/or other oxidized lipids, in the absence of constraints that are characteristic of oxidized LDL and other oxidized lipids, so they can be used to treat inflammatory diseases and disorders caused by oxidized lipids, when administered orally or through the introduction of the mucous membrane.

Since it is known that oxidized phospholipids are the active components of ox LDL, and, in addition, biological membranes typically include phospholipids, mainly phosphoglyceride, the compounds of the present invention have a structure based on oxidized phospholipids in General and oxidized phosphoglycerides in particular.

All compounds of the present invention have a General formula I

where

n is an integer from 1 to 6, and if the n=1, Cn, Bn, Rn, R n and Y are absent;

each of the B1B2, ...Bn-1and Bnindependently selected from the group consisting of oxygen, sulfur, nitrogen, phosphorus and silicon, each of the specified nitrogen, phosphorus and silicon substituted by at least one Deputy, selected from the group consisting of hydrogen, unshared pair of electrons, alkyl, halogen, cycloalkyl, aryl, hydroxy, digitoxin, alkoxy, aryloxy, diarylike, dialkoxy and oxo;

each And1And2, ...An-1and Anindependently selected from the group consisting of CR"R"', C=O and C=S;

Y is chosen from the group consisting of hydrogen, alkyl, aryl, cycloalkyl, carboxy, sugar, phosphoric acid, phosphorylcholine, phosphorylethanolamine, phosphorylcholine, phosphorylethanolamine, fosfaurilirutisa, ethylphosphonic, phosphorylmethyl, phosphorylates, fosforilirovaniya, phosphorylmethyl, phosphorylethanolamine-N-lactose, phosphoethanolamine-N-[methoxy(propylene glycol)], postinst-4-phosphate, postinst-4,5-biphosphate, pyrophosphate, phosphatidylethanolamine, dinitrobenzenamine and fosfoglitserin; and

each of X1, X2,..., Xn-1independently is a saturated or unsaturated hydrocarbon of the General formula II

where

m is an integer from 1 to 26; and

Z the choice is up from the group, consisting of

where

W is chosen from the group consisting of oxygen, sulfur, nitrogen and phosphorus, and each of the specified nitrogen and phosphorus substituted by at least one Deputy, selected from the group consisting of hydrogen, unshared pair of electrons, alkyl, halogen, cycloalkyl, aryl, hydroxy, digitoxin, alkoxy, aryloxy, diarylike, dialkoxy and oxo; and

at least one of X1, X2,..., Xn-1Z is not hydrogen;

and where

each of R1, R'1, R2, ...Rn-1, Rn, R'neach of R" and R"' and each of Ra, R'a, Rb, R b, ...Rm-1, R'm-1, Rmand R'mindependently selected from the group consisting of hydrogen, alkyl, alkenyl, quinil, cycloalkyl, aryl, heteroaryl, halogen, trihalomethyl, hydroxy, alkoxy, aryloxy, digitoxin, dialkoxy, diarylike, phosphonate, phosphate, phosphinyl, sulfonyl, sulfinil, sulfonamida, amide, carbonyl, thiocarbonyl, C-carboxy, O-carboxy, C-carbamate, N-carbamate, C-dicarboxy, S-dicarboxy and amino, or alternatively at least two of R1, R'1, R2, ...Rn-1, Rnand R'nand/or at least two of Ra, R'a, Rb, R b, ...Rm-1, R'm-1, Rmand R'mform at least one four-, five - or six-membered aromatic, hetaeras is aromatic, alicyclic or heteroalicyclic ring; and

each of the1With2, ...Cn-1Cnand each of CA, Cb, ...Cm-1and Cmis chiral or organization of the achiral carbon atom, and each chiral carbon atom has the S-configuration and/or R-configuration;

and include pharmaceutically acceptable salt, prodrug, hydrate or MES.

The specialist in this area should be clear that the possibility of introducing all of the substituents (e.g., R1-RnRa-Rm, R", R"') these provisions depends on the valency and chemical compatibility substituent, the substituted position and other substituents. Therefore, the scope of the present invention includes all possible substitute for any provisions.

Used here is the term “alkyl” means a saturated aliphatic hydrocarbon, including groups with straight and branched chain. The alkyl group preferably has 1-20 carbon atoms. The specified numeric interval, for example, “1-20” means that the group, in this case the alkyl group, may contain 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc. up to 20 carbon atoms inclusive. More preferably, the alkyl has an average chain length ranging from 1 to 10 carbon atoms. Most preferably, except where otherwise stated, the alkyl I have is lower alkyl, comprising 1-4 carbon atoms. The alkyl group may be substituted or unsubstituted. In the case of substitution of a replacement group may be, for example, cycloalkyl, alkenyl, quinil, aryl, heteroaryl, heteroalicyclic group, halogen, hydroxy, alkoxy, aryloxy, digitoxin, dialkoxy, diarylike, sulfinil, sulfonyl, cyano, nitro, azide, sulfonyl, sulfinil, sulfonamide, phosphonyl, phosphinyl, oxo, carbonyl, thiocarbonyl, urea, thiourea, O-carbamyl, N-carbamyl, thiocarbamyl, N-thiocarbamyl, C-amido, N-amido, C-carboxy, O-carboxy, sulfonamide and amino in these values.

“Cycloalkyl” group means a group with a monocyclic or condensed ring consisting of carbon atoms (i.e. with rings that share an adjacent pair of carbon atoms), in which one or more of the rings does not have a fully conjugate system of PI electrons. Non-limiting examples cycloalkyl groups include cyclopropane, CYCLOBUTANE, cyclopentane, cyclopentene, cyclohexane, cyclohexadiene, Cycloheptane, cycloheptatrien and adamantane. Cycloalkyl group can be substituted or unsubstituted. In the case of substitution of a replacement group may be, for example, alkyl, alkenyl, quinil, aryl, heteroaryl, heteroalicyclic group, halogen, hydroxy, alkoxy, aryloxy, digitoxin, tialc the XI, diarylike, sulfinil, sulfonyl, cyano, nitro, azide, sulfonyl, sulfinil, sulfonamide, phosphonyl, phosphinyl, oxo, carbonyl, thiocarbonyl, urea, thiourea, O-carbamyl, N-carbamyl, thiocarbamyl, N-thiocarbamyl, C-amido, N-amido, C-carboxy, O-carboxy, sulfonamide and amino in these values.

“Alchemilla” group means an alkyl group that contains at least two carbon atoms and at least one carbon-carbon double bond.

“Alchemilla” group means an alkyl group that contains at least two carbon atoms and at least one carbon-carbon triple bond.

“Aryl group” means a group with a monocyclic or condensed ring consisting of carbon atoms (i.e. with rings that share an adjacent pair of carbon atoms), which is a fully conjugate system of PI electrons. Non-limiting examples of aryl groups include phenyl, naphthalenyl and anthracene. The aryl group may be substituted or unsubstituted. In the case of substitution of a replacement group may be, for example, alkyl, alkenyl, quinil, cycloalkyl, aryl, heteroaryl, heteroalicyclic group, halogen, hydroxy, alkoxy, aryloxy, digitoxin, dialkoxy, diarylike, sulfinil, sulfonyl, cyano, nitro, azide, sulfonyl, sulfinil, sulfone is ID, phosphonyl, phosphinyl, oxo, carbonyl, thiocarbonyl, urea, thiourea, O-carbamyl, N-carbamyl, thiocarbamyl, N-thiocarbamyl, C-amido, N-amido, C-carboxy, O-carboxy, sulfonamide and amino in these values.

“Heteroaryl group” means a group with a monocyclic or condensed ring (i.e. with rings that share an adjacent pair of atoms), which contains in the ring one or more atoms, such as, for example, nitrogen, oxygen and sulfur, and optionally includes a fully conjugate system of PI electrons. Non-limiting examples of heteroaryl groups include pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrimidine, quinoline, isoquinoline and purine. Heteroaryl group may be substituted or unsubstituted. In the case of substitution of a replacement group may be, for example, alkyl, alkenyl, quinil, cycloalkyl, aryl, heteroaryl, heteroalicyclic group, halogen, hydroxy, alkoxy, aryloxy, digitoxin, dialkoxy, diarylike, sulfinil, sulfonyl, cyano, nitro, azide, sulfonyl, sulfinil, sulfonamide, phosphonyl, phosphinyl, oxo, carbonyl, thiocarbonyl, urea, thiourea, O-carbamyl, N-carbamyl, thiocarbamyl, N-thiocarbamyl, C-amido, N-amido, C-carboxy, O-carboxy sulfonamide and amino in these values.

“Heteroalicyclic” group SN which includes a group monocyclic or condensed ring, which contains in the ring one or more atoms such as nitrogen, oxygen and sulfur. These rings may also have one or more double bonds. However, such rings are not completely Kongeramo system PI-electrons. Heteroalicyclic group can be substituted or unsubstituted. In the case of substitution of a replacement group can be, for example, the unshared pair of electrons, alkyl, alkenyl, quinil, cycloalkyl, aryl, heteroaryl, heteroalicyclic group, halogen, hydroxy, alkoxy, aryloxy, digitoxin, dialkoxy, diarylike, sulfinil, sulfonyl, cyano, nitro, azide, sulfonyl, sulfinil, sulfonamide, phosphonyl, phosphinyl, oxo, carbonyl, thiocarbonyl, urea, thiourea, O-carbamyl, N-carbamyl, thiocarbamyl, N-thiocarbamyl, C-amido, N-amido, C-carboxy, O-carboxy, sulfonamide and amino in these values. Typical examples are piperidine, piperazine, tetrahydrofuran, tetrahydropyran, morpholine and the like.

“Hydroxyl group” means the group-HE.

“Azide” refers to the group-N=N.

“CNS” group means an-O-alkyl and-O-cycloalkyl group for these values.

“Aryloxy group” means-O-aryl and-O-heteroaryl group in these values.

“Digitately group” means the group-SH.

“Dialkoxy is owned group means-S-alkyl group, and-S-cycloalkyl group for these values.

“Diarylike” group means a-S-aryl and-S-heteroaryl group in these values.

“Carbonyl group” means the group-C(=O)-R, where R is hydrogen, alkyl, alkenyl, cycloalkyl, aryl, heteroaryl (associated carbon atom in the ring) or heteroalicyclic group (associated carbon atom in the ring) for these values.

“Aldehyde” group means a carbonyl group, in which R is hydrogen.

“Thiocarbonyl” group means the group-C(=S)-R in which R has the values defined here.

“C-carboxy” group refers to the group-C(=O)-O-R, in which R has the values defined here.

“O-carboxy” group refers to the group RC(=O)-O-, in which R has the values defined here.

“Oxo” group means a group =O.

Group “carboxylic acid” means-carboxyl group, in which R is hydrogen.

“Halogen” means fluorine, chlorine, bromine or iodine.

“Trihalomethyl group” means the group-CX3in which X means a halogen in these values.

“Sulfonylurea group” means the group-S(=O)-R in which R has the values defined here.

“Sulfonylurea group” means the group-S(=O)2-R, in which R has the values defined here.

“S-sulphonamido group” means the group-S(=O)2-NR2in which R has the shown value.

“N-sulphonamido group” means the group RS(=O)2-NR, in which R has the values defined here.

“Carbonilla group” means the group-OC(=O)-NR2in which each of R have the values defined here.

“N-carbonilla” group means a group ROC(=O)-NR-, in which each of R have the values defined here.

“Thiocarbamyl group” means the group-OC(=S)-NR2in which R has the values defined here.

“N-thiocarbamyl” group means a group ROC(=S)NR-, where each R has the values defined here.

“Amino group” means the group-NR2in which each of R have the values defined here.

“C-amido” group refers to the group-C(=O)-NR2in which each of R have the values defined here.

“N-amido” group refers to the group RC(=O)-NR-, in which each of R have the values defined here.

“Urea” refers to the group-NRC(=O)-NR2in which each of R have the values defined here.

“Guanidino group” means the group-RNC(=N)-NR2in which each of R have the values defined here.

“Guanella group” means the group R2NC(=N)in which each of R have the values defined here.

“Nitro group” means the group-NO2.

“Cyano group” means the group-C≡N.

The term “phosphonyl” or “phosphonate” refers to the group-P(=O) (OR) 2in which R has the above values. The term “phosphate” refers to the group-O-P(=O)(OR)2in which each of R have the above values.

“Phosphoric acid” means a phosphate group, in which each of R means hydrogen.

The term “phosphinyl” means the group-PR2in which each of R have the above values.

The term “thiourea” means the group-NR-C(=S)-NR-, in which each of R have the above values.

The term “saccharide” means one or more units of sugar, namely the level of sugar with an open-chain or cyclic sugar level (for example, links on the basis of pyranose or furanose), and includes any monosaccharide, disaccharide and oligosaccharide except where otherwise noted.

As can be seen from the above General formula I, the compounds of the present invention have a backbone consisting of 1-6 carbon atoms, in which at least one of these carbon atoms covalently attached to a hydrogen atom, a hydrocarbon group (alkyl, aryl, etc.), carboxyl group (for example, acyl, carboxylic acid, etc. or phosphoryl group (which is also known as a phosphate group or simply phosphate) and other 1-5 carbon atoms backbone covalently attached to a hydrocarbon chain (X1-Xn-1with the help of heteroatom (1-Bnin you is privedennoi of General formula (I). These hydrocarbon chain may include saturated or unsaturated, substituted or unsubstituted chain, optionally interrupted aromatic, alicyclic, heteroalicyclic and/or heteroaromatic parts listed in this description of the invention and shown in General formula II, whereby at least one of these chain ends oxidized group defined as above and Z are not hydrogen.

Used here is the term “hydrocarbon” means a compound that consists of hydrogen atoms and carbon atoms, covalently associated with each other. In a saturated hydrocarbon-all Sa-Cmcovalently attached to the adjacent atoms by using a simple Sigma-bond. Unsaturated hydrocarbon of at least two neighboring atom CA-Cmconnected to each other by a double bond or triple bond.

All hydrocarbon chains of the present invention may include from 1 to 26 carbon atoms, more preferably from 3 to 26 carbon atoms. Hydrocarbon chain with a terminal of the oxidized group Z may have a lower length and preferably include from 3 to 10 carbon atoms, more preferably from 3 to 6 carbon atoms excluding carbon atoms in the oxidized group.

LDL is the lipoprotein consisting of functionally different parts (components is). Among these parts are the phospholipids, which are known to play an important role in the effects of oxidized LDL on diseases associated with the formation of plaques.

Used here is the term “part” or “component” means the main part of the functionally active molecule that is associated with another molecule while maintaining its activity. Phospholipids are natural substances containing at the end of the chain nonpolar lipid group, and a highly polar vospitatelnuyu group. The most common phospholipids in nature are phosphoglyceride that consist of a glycerol backbone with attached acyl parts of fatty acids. Phosphoglyceride, such as acylphosphatase 1,2-O-fatty acids and their oxidized modifications such as POVPC and PGPC, have been used in studies of atherogenesis, as it was described above.

In different biological processes such as inflammation, in addition to LDL, phospholipids, and phosphoglycerides, participate, and other lipids. Such lipids include, for example, sphingolipids, glycolipids and other membrane lipids.

The above compounds of the present invention is created mainly on the basis of the structure of phosphoglycerides, in accordance with which in the preferred embodiment, izaberete is ia n in the General formula I is 3. Such compounds are defined in this description of the invention as analogs of oxidized phosphoglyceride, while all of the compounds of the present invention are defined as oxidized lipids and include their analogs and derivatives.

Used here is the term “analogs” refers to compounds that are structurally related to the target molecule (e.g., oxidized phospholipids, oxidized LDL, and so on) and therefore may have the same biological activity.

The term “derivative” means target molecules that have been chemically modified, but remains the main part, for example, target molecules, which were replaced by the additional or different substituents, the target molecules, some of which has been oxidized or hydrolyzed, and the like.

Given the volatility inherent acyl groups of fatty acids in natural phosphoglyceride and other structurally related compounds, which occurs due to high exposure to rapid hydrolysis in biological systems under the action of phospholipase A1(see, for example, “A Textbook of Drug Design and Development”, Povl Krogsgaard-Larsen and Hans Bundgaard, eds., Harwood Academic Publishers, chapter 13, pages 478-480), compounds of the present invention were created with the inclusion of at least one part of the simple ester O-IRN the x acids, as shown in the above General formula I, in which, when n is 3, at least one of A1and a2preferably is a group of CR"R"'. Compounds in which one of the A1and a2is a group of CR"R"'are defined in this description of the invention as monoesterification analogues of phosphoglyceride, while compounds in which each of A1and a2is CR"R"'are defined as diesterification analogues of phosphoglyceride and are characterized by improved stability in vivo, especially in comparison with the presently known synthetic oxidized phosphoglyceride (for example, POVPC and PGPC).

As shown in the above General formula I, when n is 3, at least one of X1and X2is a hydrocarbon chain that ends with an oxidized group, when Z is not hydrogen. However, as in the natural derivatives of oxidized LDL chain oxidized alkyl is typically located in the second position and, as installed, the biological activity of several phospholipids directly depends on its structure (see “prior art” for details on this issue), in another preferred embodiment of the present invention X2is a hydrocarbon chain, which ends on islenos group.

As further shown in the above General formula II, oxidized group can be, for example,

(aldehyde),

(carboxylic acid) and

(acetal), and their derivatives, such as, for example, any carboxyl or thiocarboxamide derived (for example, ester carboxylic acids, in which W stands for oxygen, and R means alkyl, aryl, cycloalkyl and similar groups), imino-derivatives (in which W means nitrogen atom), aminophosphonate (in which W stands for oxygen, and R" means Amin), phosphine or phosphonate derivatives, and many others that have been described above.

One example of a new esterified oxidized phosphoglyceride of the present invention is a simple ester of 2,5'-allegedlly (ALLE): 1-hexadecyl-2-(5'-oxapentane)-sn-glycero-3-phosphocholine (D-ALLE), 3-hexadecyl-2-(5'-oxapentane)-sn-glycero-1-phosphocholine (L-ALLE) and their racemic mixture, synthesis and application of which is described in more detail in the following section “Examples”.

However, since it is known that aldehydes are unstable compounds that are easily oxidized, preferred examples of new esterified oxidized phosphoglycerides of the present invention include acid is produced in the water 1-hexadecyl-2-(5'-carboxybutyl)-sn-glycero-3-phosphocholine (also referred to hereafter as IC-201) and its corresponding acetals 1-hexadecyl-2-(5',5'-dimethoxyphenoxy)-sn-glycero-3-phosphocholine and 1-hexadecyl-2-(5',5'-deoxypentose)-sn-glycero-3-phosphocholine (see figure 10 for exploring two-dimensional structural formula), synthesis and application of which is described in more detail in the following section “Examples”.

In addition to the above oxidized lipids isolated from phosphoglycerides, the scope of the present invention also includes oxidized lipids, selected, for example, from sphingolipids. Analogues in the form of oxidized sphingolipids of the present invention have the above-mentioned General formula I, in which n is 3, Y represents hydrogen, In2mean NH and a2means C=O, with a hydrocarbon chain with a terminal of the oxidized group attached to the amide, denoted by X2or With1.

Although the oxidized phosphoglyceride selected from glycerol, which is a monosaccharide molecule, and oxidized sphingolipids isolated from sphingosine, amerosport, we can assume that oxidized phospholipids isolated from other biologically widespread structural units of alcohol have the same effect. In addition, since no correlation was observed between the distance of the location of the oxidized part and vospalitelnoe part in oxidized phospholipids, it is believed that oxidized lipids isolated from the frame, comprising 4-6 carbon atoms, must retain structural characteristics similar to the characteristics of oxidized FOSFA is of glycerides and thus, should have the same antigenicity and immunomodulatory activity, so that they can be used similarly derived oxidized phosphoglycerides discussed in this description.

A preferred example of such a structural element of alcohol is a structural element of a monosaccharide, such as glucose, aritra and trait.

In another preferred variant of the invention, the compounds of the present invention wcscat to 6 carbon atoms in the chain backbone. The carbon atoms in the chain backbone can be linearly attached to each other with the formation of the skeleton of a monosaccharide with an open circuit or alternative education heteroalicyclic backbone of monosaccharide, namely the skeleton of the pyranose or furanose, while in the above General formula is one of R1and R'1covalently attached to one of Rnor R'nsimple essential communication (communication R-O-R).

The alternate connection of the present invention may contain 4-6 carbon atoms in the chain backbone, which form neskorena ring, namely, four-, five - or six-membered carbon or heteroalicyclic ring, while in the above General formula I one of R1and R'1covalently attached to one of Rnor R'ndifferent light is s (for example, Sigma-bond π-bond, carboxyl link, simple ether bond, thioester bond, and any other communications).

As further shown in the above General formula I, Y represents the phosphoryl part (for example, phosphorylcholine, phosphorylethanolamine etc) or neosvoennoy part (for example, hydrogen, acyl or alkyl). When Y means neosvoennoy part, the compound obtained is not a phospholipid, and a connection diglyceride, for n=3, or any other compound derived from alcohol, for example a compound selected from a monosaccharide.

So as not reported any activity phosphoryl group in respect of the immunomodulatory activity of oxidized LDL, it can be assumed that such neosporine connection must retain structural characteristics similar to the above-mentioned oxidized phospholipids, therefore, must have antigenicity and immunomodulatory activity and can therefore be used similarly derived oxidized phospholipid discussed in this description.

In one embodiment of the present invention, Y represents a saccharide as described above, and thus, the connection of the present invention is oxidized analogue glycolipids.

In another embodiment of the invention the connection t is aetsa oxidized analogue of any of membrane lipid.

The preferred structural features discussed above for oxidized phosphoglycerides apply to all of the above compounds. Therefore, in a preferred embodiment of the present invention, at least one And1And2... and An-1is a group of CR"R"', and the connection includes at least one esterified side chain. Given the instability of the O-acyl side chain, it is desirable that at least one of the oxidized groups in the X1-Xn-1was associated with such esterified side chain.

Although natural phospholipids and oxidized phospholipids typically include the O-acyl chains, there is evidence suggesting that thiol derivatives of oxidized phospholipids, which include, for example, S-acyl chains may have the same biological activity (see, for example, publications Reddy et al. Antitumor ether lipids: an improved synthesis of ilmofosine and an entioselective synthesis of an ilmofosine analog. Tetrahedron Letters. 1994;17:2679-2682; Batia and Hajdu. Stereospecific synthesis of ether and thioether phospholipids. The use of L-glyceric acid as a chiral precursor phospholipids. J. Org. Chem. 1988;53:5034-5039; Bosies et al. Lipids. 1987;22:947; Bosies et al. Ger. Offen. DE 3,906,952 [C.A. 1991, 114,102394w]; and Herrmann et al. NCI-EORTC Symposium on New Drugs in Cancer Therapy, Amsterdam, March 1992]. Because thiols characterized high biological stability, such compounds can be very useful.

In one embodiment, the done by the means of the present invention at least one of In 1-Bnmeans sulfur, with at least one of the side chains is etiolirovannoy S-acyl, or S-alkyl chain. In another embodiment of the invention at least one of X1-Xn-1including oxidized group, associated with such etiolirovannoy side chain.

Alternative every one of In1-Bnmay represent a biologically compatible heteroatom, which is not an oxygen atom, and sulfur, such as, for example, nitrogen, phosphorus or silicon, as shown in the above General formula I.

In addition to the structural features discussed in this description of the invention, the compounds of the present invention can be further substituted in any position, for example, any carbon atom in the side chain or at any carbon atom in the skeleton. Although there are many possible substituents described above that are included in the scope of the present invention, preferred substituents are, for example, halogen and aryl.

Although the compounds of the present invention were created on the basis of oxidized phospholipids, such as phosphoglyceride, the inventors believe that a simple oxidized hydrocarbon chain, which is optionally attached to a polar group, must have the same antigenicity and immunomodulary activity, as described above, the s analogs of oxidized phospholipid.

Such oxidized uglevodorodnaya chain is a common feature of the metabolites of arachidonic acid. Arachidonic acid is a polyunsaturated fatty acid with 20 carbon atoms, which is formed in vivo by enzymatic hydrolysis of phospholipids containing this acid. Selected arachidonic acid is oxidized in a number of important physiologically active substances defined lipoxygenase, and after a number of additional enzymatic reactions these physiologically active substances are metabolized in the family of classical prostaglandins (PG), prostacyclins (PGI2) and thromboxanes (TX) And2that are active in many biological pathways exchange. All of these metabolites have a common characteristic, which is the chain of six carbon atoms and ending with oxidizable double bonds.

As indicated above and will be further demonstrated in the Examples section, the importance of the presence of oxidized groups in analogs of oxidized LDL, which are designed to simulate the immunomodulation induced by ox LDL. Thus, in comparative studies, for example, it was shown that the connection V, oxygenated compound, the corresponding CI-201 (compound VII)is inactive, while CI-201 is active (see, for example, note the ry XIV and XV in the following “Examples”section). In addition, on the basis of route of metabolism of arachidonic acid can be assumed that other oxidized phospholipids are the same way exchange, resulting in the selection of the oxidized side chain. As will be described below, the oxidized side chain preferably contains from 3 to 7 carbon atoms and therefore a similar chain of six carbon atoms, metabolites of arachidonic acid. In addition, the above mechanism CD1, which includes involvement of lipids in the immune system, shows that the hydrophilic head group, that is, the parent group of a carbon-C2 and/or carbon-C3 in CD1-d, is most likely antigenic determinant, we present the immune system, as it is part represented by the groove CD1, which hides the hydrophobic portion of the molecule, indicating a role hydrophilic epitope in the position of the carbon-C2.

In oxidized phospholipids, such as phosphoglyceride, oxidized side chain attached to phosphoglyceromutase the core. However, as stated above, phosphoglycerides skeleton doesn't have any particular role.

In a preferred embodiment of the present invention n is 1, the compound of the present invention has a simple hydrocarbon chain with a terminal of the oxidized group. Although this okislennya is a simple hydrocarbon chain is non-polar, it can be attached to a polar group, such as phosphoryl group, in this case, as shown in the above General formula I, when n is 1, at least one of R1and R'1is a phosphate or phosphonate group. The alternative at least one of R1and R'1you can choose from other biocompatible polar groups, such as, for example, peptides, saccharides and the like.

Depending on the substituents, all the carbon atoms in the above compounds, namely1-Cnand Sa-Cm, can be chiral or organization of the achiral. Any chiral carbon atom present in the compounds of the present invention, may have the R-configuration, S-configuration, or to be racemic. Thus, the scope of the present invention includes any combination of chiral and racemic carbon atoms, including all possible stereoisomers, optical isomers, enantiomers and anomer. As shown in the following section the Examples, the compounds of the present invention can be synthesized while maintaining the configuration of the original substance. Compounds of the present invention can be further selectively synthesized with stereochemical details of oxidized groups. In selecting appropriate starting compounds and the appropriate synthesis conditions can determine the shape of the optical purity (for example, the inclusion of chiral and/or racemic carbon atoms) and the stereoisomers obtained soedinenii. In the case of obtaining racemic mixtures, it is possible to use known methods for separation of optical or stereoisomers. Such methods are described, for example, in the publication “Organic chemistry, fourth Edition by Paula Yurkanis Bruice, page 180-185 and page 214, Prentice Hall, Upper Sadde River, NJ 07458”.

In the scope of the present invention further includes any pharmaceutically acceptable salt, prodrug, hydrate and solvate of the above compounds.

The term “prodrug” means a vehicle that turns into active connection (active original medicinal substance) in vivo. Prodrugs are usually used to facilitate the introduction of the original medicinal substance. Prodrugs, for example, can be biologically suitable for oral administration, while the original medicinal substance may not be suitable for such an introduction. The prodrug may also have better solubility compared to the original medicinal substance in pharmaceutical compositions. In addition, prodrugs often use for prolonged release of the active soedineniya in vivo. Non-limiting examples of prodrugs may be the compound of the present invention, having one or more parts of the carboxylic acid, which BB is car Ried out in the form of ester (the“prodrug”). Such prodrug is hydrolyzed in vivo to the formation of free connection (original medicinal substance). Selected ester may affect both the characteristics of solubility and hydrolysis rate of the prodrug.

The term “pharmaceutically acceptable salt” means a prewired kind of parent compound and its counterion, which is usually used to modify the characteristics of solubility of the parent compound and/or to reduce the irritation caused by the body-source connection, without altering the biological activity and properties of the input connections. Non-limiting examples of pharmaceutically acceptable salts can be a carboxylate anion and a cation, which includes, but is not limited to, ammonium, sodium, potassium and the like.

The term “MES” means a complex of variable stoichiometry (e.g., di-, tri-, Tetra-, Penta-, hexa and so on), which is formed of dissolved substance (compound of the present invention) and a solvent, whereby the solvent should not affect the biological activity of the solute. Acceptable solvents include, for example, ethanol, acetic acid and the like.

The term “hydrate” means the above MES, in which the solvent is water.

As will be described in detail below, but what's the connection of the present invention have a high immunomodulatory activity and can therefore be used in different therapeutic applications. The use of these compounds in therapeutic applications include the introduction of these compounds per se or in the form of pharmaceutical compositions, in which such a compound is mixed with an acceptable carriers or excipients.

Thus, another object of the present invention is a pharmaceutical composition which contains as an active ingredient any of the above compounds of General formula I and a pharmaceutically acceptable carrier.

Used here is the term “pharmaceutical composition” means a product containing one or more of the above active ingredients, together with other chemical components such as physiologically acceptable carriers and excipients. The aim of creating a pharmaceutical composition is to facilitate the introduction of the compound into the body of the subject.

The term “active ingredient” means a compound (e.g., ALLE, CI-201 and other compounds corresponding to the above General formula (I)possess biological activity.

The terms “physiologically acceptable carrier” and “pharmaceutically acceptable carrier”, which have interchangeable values, indicate carrier or a diluent that does not cause significant irritation to an organism and does not affect the biological activity and properties the and input connections. Adjuvant is included in the definition of these terms.

The term “filler” means an inert substance added to a pharmaceutical composition to further facilitate the introduction of the active ingredient. Non-limiting examples of fillers include calcium carbonate, calcium phosphate, various sugars and starches, cellulose derivatives, gelatin, vegetable oils and polyethylene glycols.

Methods of obtaining and administering drugs described in the publication “Remington''s Pharmaceutical Sciences,” Mack Publishing Co., Easton, P.A., latest edition, which is incorporated in this description by reference.

Acceptable routes of administration can include, for example, oral and rectal administration, the introduction through the mucous membrane, especially in the nose, intestinal or parenteral administration, including intramuscular, subcutaneous and intramedullary injections as well as intrathecal, direct intraventricular, intravenous, intraperitoneal, nasal, or intraocular injections.

Alternative pharmaceutical composition can be entered not systemically and locally, for example, in the form of injection of the pharmaceutical composition directly into the affected tissue of the subject.

In a preferred embodiment of the present invention the pharmaceutical compositions are used to modulate the immune and/or vocal the positive reactions when introduced through the mucous membrane.

In another preferred embodiment of the present invention the pharmaceutical composition is intended for modulation of the immune and/or inflammatory response when administered orally.

The pharmaceutical compositions of the present invention is preferably intended for nasal or intraperitoneal injection, as described in detail below.

The pharmaceutical compositions of the present invention can be obtained by methods well known in this field, for example by mixing, dissolving, granulating, production drops, grinding into powder, emulsifying, encapsulating, wrapping or lyophilization.

Pharmaceutical compositions intended for use in the present invention, can be obtained in the usual way using one or more physiologically acceptable carriers comprising excipients and auxiliaries, which facilitate processing of the active ingredents drugs and can be used in pharmaceutical. A particular drug depends on the selected method of administration.

To obtain injectable active ingredients of the pharmaceutical compositions can be obtained in aqueous solutions, preferably in physiologically compatible buffers, such is how the solution Hank, the ringer's solution or fiziologicheskii solution. For insertion through the mucous membrane of the use of penetrating substances corresponding to overcome the barrier. Such penetrating substances is well known in this field.

Pharmaceutical composition for oral administration can be obtained by combining the active compounds with pharmaceutically acceptable carriers well known in this field. Such carriers enable you to obtain a pharmaceutical composition in the form of tablets, pills, coated tablets, capsules, liquids, gels, syrups, slurries, suspensions and the like, which are intended for ingestion by the subject. Pharmaceutical preparations for oral administration can be obtained using a solid excipient, optionally grinding the resulting mixture and produce pellets of a specified mixture after adding, if desired, acceptable auxiliary substances for the manufacture of tablets or core tablets. Acceptable excipients are, in particular, sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as, for example, corn starch, wheat starch, rice starch, potato starch, gelatin, tragant, methylcellulose, hypromellose, sodium-carbometalation; and/or physiologically when mimie polymers, such as polyvinylpyrrolidone (PVP). If desired, can be added substances that improve raspadaemost, such as cross-crosslinked polyvinylpyrrolidone, agar, alginic acid or its salt, such as sodium alginate.

At the core of pills cause reasonable coverage. For this purpose you can use concentrated sugar solutions, which may not necessarily contain the Arabian gum, talc, polyvinylpyrrolidone, gel, carbopol, polyethylene glycol, titanium dioxide, solutions varnish and acceptable organic solvents or solvent mixtures. In tablets or dragee coatings for identification or to characterize the different combinations of doses of active compounds can be added dyes or pigments.

Pharmaceutical compositions which can be used orally include hard capsules made of gelatin, and also soft sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. The hard capsules can contain the active ingredients in a mixture with fillers, such as lactose, binders, such as starch, and lubricating agents such as talc or magnesium stearate, and optionally a stabiliser. In soft capsules, the active ingredients can be dissolved or suspended in an acceptable l is bidah, such as fatty acids, liquid paraffin, or liquid polyethylene glycols. In addition, can be added stabilizers. All preparations for oral administration should be in a unit dosage form suitable for the selected method of administration.

Compositions for transbukkalno introduction can be in the form of tablets or pellets obtained in the usual way.

Medicines for inhalation through the nose, containing the active ingredients of the present invention, is usually administered in the form of an aerosol spray from a container under pressure, or a nebulizer with the use of acceptable propellant, such as DICHLORODIFLUOROMETHANE, Trichlorofluoromethane, dichlorotetrafluoroethane or carbon dioxide. In the case of a pressurized aerosol, the device may be provided with a valve for supplying a measured quantity. Capsules and cartridges manufactured, for example, from gelatin, which are intended for use in the dosing device may contain a powder mix of the compound and acceptable powder base such as lactose or starch.

The pharmaceutical composition of the present invention may be intended for parenteral administration, such as injection loading dose of the substance or continuous influence. Preparations for injection can be obtained in the metered addition to the state form, for example in ampoules or containers designed for multiple doses, with optional added preservative. These compositions can be represented as suspensions, solutions or emulsions in maslanyj or aqueous diluents, and may include forming agents, such as suspendida, stabilizing and/or dispersing agents.

Pharmaceutical compositions for parenteral administration include aqueous solutions of the active drug in a water soluble form. In addition, suspensions of the active ingredients can be obtained in the form of injectable suspensions in oil or water-based. Acceptable lipophilic solvents or carriers include fatty acids, such as sesame oil, or synthetic fatty acids, such as etiloleat, triglycerides, or liposomes. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. The slurry may optionally contain acceptable stabilizers or agents that increase the solubility of the active ingredients, the result can be obtained in highly concentrated solutions.

Alternatively, the active ingredient may be in powder form, intended for dilution immediately before use in an acceptable solvent is, such as, for example, sterile pyrogen-free aqueous solution.

The pharmaceutical composition according to the present invention can be also obtained in the form of compositions for rectal administration, such as suppositories or retention enema, using, for example, conventional bases for suppositories, such as cocoa butter and other glycerides.

The pharmaceutical compositions of the present invention include compositions where the active ingredients are present in an effective amount sufficient to achieve the objectives. In particular, a therapeutically effective amount refers to an amount of active ingredients, which can effectively prevent, eliminate or reduce the symptoms of the disease (such as atherosclerosis) or extend the life of the subject being treated.

Specialists in this field can easily identify therapeutically effective amount, particularly given the detailed description of the invention.

For any preparation used in the implementation of the methods according to this invention, a therapeutically effective amount or dose can be calculated first by performing the in vitro and exploring the culture of cells. For example, the dose can be determined using animal models, allowing to establish the desired concentration or t is R. Such information can be used for more accurate determination of doses suitable for treatment of humans.

Toxicity and therapeutic efficacy of the active ingredients discussed in this description of the invention, it is possible to define a standard pharmaceutical methods in vitro using cell cultures or experimental animals. Data obtained from these analyses in vitro studies of cell cultures and animals, can be used to determine the range of doses that are suitable for the treatment of humans. The dose may vary depending on the dosage form and route of administration. Exact drug, route of administration and dosage can be chosen by the attending physician taking into account the state of need of the subject (see, for example, the publication Fingl, et al., 1975, in “The Pharmacological Basis of Therapeutics, Ch.1, p.1).

The largest doses and intervals between doses of the medicinal product can be defined individually for achievements in plasma or brain levels of the active ingredient sufficient for the induction or suppression of angiogenesis (minimal effective concentration, MONTHS). The minimum effective concentration may be different for each drug, but may be determined on the basis of data obtained in vitro. The dose required to achieve the minimum effective concentration is AI, depend on individual characteristics and route of administration. For determination of the concentration in plasma is possible to use known methods of detection.

Depending on the severity of the disease and response to treatment, it is possible to produce single or multiple dose, and the treatment can last from several days to several weeks, until full recovery or mitigate symptoms of the disease.

The number of input composition, no doubt, depends on the state of the subject in need of treatment, the severity of the lesion, the method of administration, the decision of the attending physician, etc.

The compositions of the present invention optionally can be in the packaging or desirudin device, such as set and approved by Management under the control over products and medicines (FDA), which may include one or more dosage forms containing the active ingredient. Packing can be a metal or plastic foil, such as a blister pack. To the packaging or the dosing device may be accompanied by instructions for administration. Packing or Desiree device may also be provided with a notification corresponding to the form prescribed by a governmental Agency regulating the manufacture, use or sale of pharmaceutical PR the preparations which should contain information about the authorization data by the Agency of the form of the compositions or fitness for a human or animal. Such a notification may, for example, be marked, approved by Management under the control over products and medicines of the USA for their medication as prescribed, or the liner about the suitability. In addition, there can be obtained a composition containing the drug of the present invention together with a compatible pharmaceutical carrier, which is placed in an appropriate container and labeled with an indication of disease, treatment for which they are intended, as will be described in detail below.

Thus, in a preferred embodiment of the present invention the pharmaceutical composition is packaged in a packaging material and point in a paper liner or packing material that the composition is intended for the treatment or prophylaxis of inflammation associated with endogenous oxidized lipid. A list of common diseases and disorders associated with such inflammation, below.

As will be described in more detail below, the pharmaceutical composition of the present invention may further include additional connection, suitable for the treatment or prevention of the above inflammation.

As described in detail in Nigel is blowing the “Examples”section, the new compounds of the present invention effectively modulate the immune response and/or inflammation associated with endogenous oxidized LDL, thus causing a weakening of the symptoms of diseases associated with endogenous oxidized LDL. The obtained results clearly show that (i) the immune and/or inflammatory reaction to endogenous oxidized LDL, in particular, and endogenous oxidized lipids can be modulated by any compound structurally related oxidized lipid; and (ii) compounds capable of modulating the immune and/or inflammatory response to oxidized lipids, can be used for the treatment or prophylaxis of inflammation associated with endogenous oxidized lipids.

Thus, another object of the present invention is a method of treatment or prophylaxis of inflammation associated with endogenous oxidized lipid. The method according to this object of the present invention is carried out by introducing the needy to the subject a therapeutically effective amount of one or more oxidized lipids.

Used here is the term “endogenous oxidized lipid” means one or more oxidized lipids that are present or formed in vivo in inflammatory and other processes, OPOS duemig cells or humoral system.

The term “method” means the methods, tools, techniques, and procedures for achieving these goals include, but are not limited to, methods, tools, techniques, and procedures that are known in this field or can be easily developed on the basis of known methods, means, techniques and procedures by specialists in the chemical, pharmacological, biological, biochemical and medical fields.

Used here is the term “treatment or prevention” means the removal, essentially inhibiting, slowing or reversal of disease, weakening of the clinical symptoms of the disease or prevent the clinical symptoms of the disease.

Entities that may be appointed such treatment, are subjects suffering from diseases or disorders associated with inflammation, which are described in detail below. The preferred subjects of the present invention include mammals, such as dogs, cats, sheep, pigs, horses and cattle. The subjects of the present invention preferably are people.

The term “oxidized lipid” means natural or preferably synthesized compound having the General structural features with the natural lipid oxidized lipid and any component of the AMI, parts, analogues and derivatives. For example, oxidized LDL consists of several functionally and structurally different parts, and in the definition of this term includes any of the synthesized compounds having the General structural characteristics with any of these parts. In the definition of this term further includes any derivative of such analogs.

Typical examples of oxidized lipids include, but are not limited to, oxidized phospholipids, analogs of platelet activating factor, analogs of plasmalogen, substituted or unsubstituted hydrocarbons containing 3-30 carbon atoms and leaf oxidized group, oxidized analogs of sphingolipids, oxidized analogues of glycolipids, oxidized analogues of membrane lipids and any of its analogs or derivatives.

Phospholipids in General and phosphoglyceride, in particular, are well known lipids, which are also components of oxidized LDL. Phosphoglyceride are derived fosfoglitserin, which contains one or more acyl groups of fatty acids or acyl groups attached to phosphoglyceromutase the skeleton.

Synthesized oxidized phospholipids can be used effectively in the implementation of the method according to this object of the present invention. Typical examples of known synthetic oxidized phospholipids including the indicate, not limited to, 1-Palmitoyl-2-azelaoyl-sn-glycero-3-phosphocholine, 1-hexadecyl-2-azelaoyl-sn-glycero-3-phosphocholine, 1-Palmitoyl-2-glutaryl-sn-glycero-3-phosphocholine (PGPC), 1-Palmitoyl-2-(5-oxovalerate)-sn-glycero-3-phosphocholine (POVPC) and 1-Palmitoyl-2-(9-Oconnor)-sn-glycero-3-phosphocholine.

More preferred examples of synthetic oxidized phospholipids include the above compounds of the present invention, in which n=1. These compounds defined in the present description of the invention as substituted or unsubstituted hydrocarbon containing 3-30 carbon atoms and leaf oxidized group.

Other compounds that have a structure related oxidized phosphoglycerides, and therefore can be used effectively in the implementation of this and other objects of the present invention are analogs of platelet activating factor (PAF).

PAF are 1-alkyl-2-acetyl-sn-glycero-3-phosphocholine, natural glycerolipid associated simple ether bond. The alkyl chain in position sn-1 is usually unsaturated alkyl having 16-18 carbon atoms. Some well-known analogues of PAF are usually characterized by the substitution of the acyl part in position sn-2 long-chain acyl part (for example, acyl fatty acid). Additional analogs of PAF include oxidative modification is s in position sn-1 Of unsaturated-alkyl chain or in position sn-2 acyl chain fatty acids.

Typical examples of known analogues PSF, which can be used in the present invention include, but are not limited to, 1-Palmitoyl-2-(9-Oconnor)-sn-glycero-3-phosphocholine, 1-hexadecyl-2-acetyl-sn-glycero-3-phosphocholine, 1-octadecyl-2-acetyl-sn-glycero-3-phosphocholine, 1-hexadecyl-2-buterol-sn-glycero-3-phosphocholine, 1-octadecyl-2-buterol-sn-glycero-3-phosphocholine, 1-Palmitoyl-2-acetyl-sn-glycero-3-phosphocholine, 1-octadecenyl-2-acetyl-sn-glycero-3-phosphocholine, 1-hexadecyl-2-(homogametic)-sn-glycero-3-phosphocholine, 1-hexadecyl-2-arachidonoyl-sn-glycero-3-phosphocholine, 1-hexadecyl-2-eicosapentaenoic-sn-glycero-3-phosphocholine, 1-hexadecyl-2-docosahexaenoyl-sn-glycero-3-phosphocholine, 1-octadecyl-2-methyl-sn-glycero-3-phosphocholine, 1-hexadecyl-2-butanoyl-sn-glycero-3-phosphocholine, Lyso C16 PAF and Lyso PAF C18. However, in the context of the present invention can also be used any other PAF analogues or derivatives thereof.

Additional compounds that have a structure related oxidized phosphoglycerides, and therefore can be effectively used on this and other objects of the present invention are analogs of plasmalogen.

Plasmalogen are 1-alkyl-2-acetyl-sn-glycero-3-phosphatidyl natural glycerolipid associated simple ether bond, in which the alkyl chain in position sn-1 usually is is intense. Some well-known analogues of plasmalogens usually characterized by the substitution of the acyl part in position sn-2 long-chain acyl part (for example, acyl fatty acid), and further include oxidative modification in position sn-1 or in position sn-2.

Typical examples of known analogues of plasmalogen, which can be used in the context of the present invention, include, but are not limited to, 1-On-1'-(Z)-hexadecanoyl-2-[12-[(7-nitro-2-1,3-benzoxadiazole-4-yl)amino]dodecanoyl]-sn-glycero-3-phosphocholine, 1-On-1'-(Z)-hexadecanoyl-2-oleoyl-sn-glycero-3-phosphocholine, 1-On-1'-(Z)-hexadecene-1-arachidonoyl-sn-glycero-3-phosphocholine, 1-On-1'-(Z)-hexadecanoyl-2-docosahexaenoyl-sn-glycero-3-phosphocholine, 1-On-1'-(Z)-hexadecanoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine, 1-On-1'-(Z)-hexadecanoyl-2-arachidonoyl-sn-glycero-3-phosphoethanolamine and 1-On-1'-(Z)-hexadecanoyl-2-docosahexaenoyl-sn-glycero-3-phosphoethanolamine. However, in the context of the present invention can also be used any other analogues of plasmalogens or their derivatives.

Used here is the term “inflammation associated with endogenous oxidized lipid” refers to the inflammation that is associated with the formation or presence of in vivo one or more oxidized lipids (such as oxidized LDL, oxidized membrane lipids etc).

Inflammation is the protective reaction of the body to injury. The inflammatory response is mediated by several cytokines, including IFN-γ and IL-10. IFN-γ is involved in the pathogenesis of various autoimmune and chronic inflammatory conditions. On the other hand, IL-10 inhibits the production of IFN-γ activated immunocompetent cells, such as T m M2, from which it follows that this cytokine (IL-10) is the main anti-inflammatory “shield”.

Severe inflammation often is destructive and leads to numerous diseases and disorders. As described in detail above, a strong inflammatory reaction usually associated with epitopes of oxidized lipids.

As shown in the following “Examples”section, modulation of the immune response to oxidized LDL using synthetic analogs of oxidized LDL anti-inflammatory effect. Specified anti-inflammatory action can be used to treat or prevent diseases or disorders caused by inflammation, which is associated with endogenous oxidized LDL or any other endogenous oxidized lipid. Such diseases and disorders include, for example, diseases or disorders caused by the formation of plaques, which include, but are not limited to, atherosclerosis, atherosclerotic cardiovascular disease, cerebrovascular is s disease, peripheral vascular disease, stenosis, restenosis, stenosis in the stent, autoimmune diseases or disorders, neurodegenerative diseases or disorders, proliferative diseases or disorders and the processes associated with aging.

Thus, typical examples of diseases or disorders caused by inflammation, which in turn is associated with endogenous oxidized lipids, and therefore suitable for treatment by the method according to the present invention, include, for example, idiopathic inflammatory diseases or disorders, chronic inflammatory diseases or disorders, acute inflammatory diseases or disorders, autoimmune diseases or disorders, infectious diseases or disorders, inflammatory malignant diseases or disorders, inflammatory diseases or disorders associated with transplantation, inflammatory degenerative diseases or disorders, diseases or disorders caused by hypersensitivity, inflammatory cardiovascular diseases or disorders, inflammatory cerebrovascular diseases or disorders, diseases or disorders peripheral vascular, inflammatory diseases or disorders of the glands, inflammatory gastrointestinal diseases or disorders, inflammatory skin is haunted diseases or disorders, inflammatory diseases or disorders of the liver, inflammatory neurological diseases or disorders, inflammatory diseases or disorders of the musculoskeletal system, inflammatory diseases or disorders of the kidney, inflammatory diseases or disorders of the reproductive system, inflammatory systemic diseases or disorders, inflammatory diseases or disorders of connective tissue, inflammatory tumors, necrosis, inflammatory diseases or disorders associated with the implant, inflammatory processes associated with aging, diseases or disorders caused by immune, proliferative diseases and disorders, and inflammatory pulmonary diseases or disorders, as described below.

Non-limiting examples include hypersensitivity hypersensitivity type I hypersensitivity type II hypersensitivity type III hypersensitivity type IV hypersensitivity of immediate type, antitelomerase hypersensitivity, hypersensitivity, mediated by immune complex hypersensitivity mediated by T-lymphocytes, delayed-type hypersensitivity, hypersensitivity mediated by T-lymphocytes-helper, hypersensitivity, mediated cytotoxics the mi T-lymphocytes, hypersensitivity mediated T-lymphocytes, and hypersensitivity mediated T-lymphocytes.

Non-limiting examples of inflammatory cardiovascular diseases or disorders include obtenerse diseases or disorders, atherosclerosis, valvular defect, stenosis, restenosis, stenosis in the stent, myocardial infarction, coronary heart disease, acute coronary syndromes, congestive heart failure, angina, coronary infarction, thrombosis, granulomatous's granulomatosis, Takayasu's arteritis, Kawasaki syndrome, an autoimmune disease or disorder directed against factor VIII, necrotizing vasculitis of small vessels, the microscopic disorder, syndrome charge and Strauss, legomania focal necrotizing glomerulonephritis, Crescent glomerulonephritis, antiphospholipid syndrome, antialienation heart failure, thrombotic purple, autoimmune hemolytic anemia, autoimmune heart disease, Chagas disease and autoimmunity against T-lymphocyte helper.

Stenosis is obtenerse disease vascular network, which typically occurs due to the formation of atherosclerotic plaques and increased platelet activity, and mainly affects the vascular system of the heart.

Restenosis is the progress yuusha re obturation, which often occurs after removal of obduracy in stenotic vascular system. In those cases, when the open state of the vessel may be provided with mechanical support stent can occur stenosis in the stent that causes obstruction of the restored vessel.

Non-limiting examples of cerebrovascular diseases or disorders include stroke, inflammation of the blood vessels of the brain, intracerebral hemorrhage and vertebral arterial insufficiency.

Non-limiting examples of diseases and disorders of the peripheral vessels include gangrene, diabetic vasculitis, ischemic bowel disease, thrombosis, diabetic retinopathy and diabetic nephropathy.

Non-limiting examples of autoimmune disorders include all diseases caused by an immune reaction, such as antibodies or cretaceoustertiary immune response to autoantigen and the like. Typical examples are chronic rheumatoid arthritis, of still's disease, systemic lupus erythematosus, scleroderma, a complex disease of connective tissue, Nowotny polyarteritis, polymyositis/dermatomyositis, Sjogren syndrome, syndrome behceta, multiple sclerosis, autoimmune diabetes, Hashimoto's disease, psoriasis, primary myxedema, pernicious anemia, myasthenia gravis, x is oncesi active hepatitis, autoimmune hemolytic anemia, idiopathic thrombocytic purpura, uveitis, vasculitis and heparininduced thrombocytopenia.

Non-limiting examples of inflammatory diseases or disorders of the glands include diseases or disorders of the pancreas, diabetes type I, diseases or disorders of the thyroid gland, graves ' disease, thyroiditis, spontaneous autoimmune thyroiditis, Hashimoto thyroiditis, the idiopathic myxedema, autoimmune disease of the ovary, autoimmune male infertility, autoimmune prostatitis and autoimmune pluriglandular syndrome type I.

Non-limiting examples of inflammatory gastrointestinal diseases or disorders include colitis, REIT, Crohn's disease, a chronic inflammatory disease of the small bowel, irritable bowel syndrome, a chronic inflammatory disease of the colon, coeliac disease, ulcerative colitis, ulcers, skin ulcers, bedsores, ulcers, peptic ulcer, buccal ulcers, nasal ulcer, esophageal ulcer, duodenal ulcer and ulcer of the gastrointestinal tract.

Non-limiting examples of vospaliteleny skin diseases or disorders include acne and autoimmune bullous skin disease.

Non-limiting examples of inflammatory diseases or disorders of the liver include the indicate autoimmune hepatitis, cirrhosis of the liver and biliary cirrhosis.

Non-limiting examples of inflammatory neurological diseases or disorders include multiple sclerosis, Alzheimer's disease, Parkinson's disease, myasthenia gravis, motor neuropathy, Guillain-Barre syndrome, autoimmune neuropathy, myasthenic syndrome Lambert-Eaton paraneoplastic neurological disease or disorder, paraneoplastic cerebellar atrophy, nephronophthisis syndrome stiffness, progressive cerebellar atrophy, Rasmussen encephalitis, amyotrophic lateral sclerosis, horey of Sydenham syndrome Gilles de La Tourette, the autoimmune polyendocrinopathy, neuropathy, due to impaired immunity, acquired neuromyotonia, multiple arthrogryposis, Huntington's disease, dementia caused by AIDS, amyotrophic lateral sclerosis (AML), multiple sclerosis, stroke, inflammatory disease of the retina, inflammatory eye disease, retro-bulbar neuritis, gubavica encephalopathy, migraine, headache, histamine headache syndrome and stiffness.

Non-limiting examples of inflammatory diseases or disorders of connective tissue include autoimmune myositis, primary Sjogren syndrome, an autoimmune disease or disorder of smooth muscle myositis, tendin is t, inflammation of the ligaments, leakage, inflammation of joints, synovial inflammation, syndrome carpal, arthritis, rheumatoid arthritis, osteoarthritis, ankylosing spondylitis, inflammation of the skeletal system, autoimmune ear disease and autoimmune disease of the inner ear.

Non-limiting examples of inflammatory diseases or disorders of the kidneys include autoimmune interstitial nephritis and/or kidney cancer.

Non-limiting examples of inflammatory diseases or disorders of the reproductive system include the repeated loss of the fetus, a cyst, a disease or disorder associated with the menstrual cycle.

Non-limiting examples of inflammatory systemic diseases or disorders include systemic lupus erythematosus, systemic sclerosis, septic shock, toxic shock syndrome, and cachexia.

Non-limiting examples of infectious diseases or disorders include chronic infectious diseases or disorders, subacute infectious diseases or disorders, acute infectious diseases or disorders, viral diseases or disorders, bacterial diseases or disorders, diseases or disorders caused by protozoa, parasitic diseases or disorders, fungal, mycoplasmal diseases or disorders, gangrene, sepsis, diseases or is arsenia, caused by insects, influenza, tuberculosis, malaria, acquired immunodeficiency syndrome and severe acute respiratory illness.

Non-limiting examples of inflammatory diseases or disorders associated with transplantation, include graft rejection, chronic graft rejection, subacute graft rejection, acute graft rejection, Verhoture graft rejection and graft-versus-host. Typical implants include prosthetic implant, breast implant, silicone implant, dental implant, implant penis, heart implant, artificial joint, a device for recovery of broken bones, bone implant, the implant for drug delivery, a catheter, a pacemaker, artificial heart, artificial heart valve, an implant that releases the drug, the electrode and the breathing tube.

Non-limiting examples of inflammatory tumors include malignant tumor, benign tumor, a solid tumor, metastatic tumor, and solid tumor.

Non-limiting examples of inflammatory lung diseases and disorders include asthma, allergic asthma, emphysema, chronic obtenerse lung disease, sarcoidosis and bronchitis.

Examples of proliferative diseases or disorders is a cancer.

In the scientific literature recently described the use of phospholipids and phospholipid metabolites for the treatment or prevention of diseases and syndromes, such as, for example, oxidative stress in the aging process (Onorato J.M., et al., Annal NY. Acad. Sci. 1998 Nov. 20; 854:277-90), rheumatoid arthritis (RA) (Paimela L, et al., Ann. Rheum. Dis. 1996 Aug; 55(8):558-9), a disease of Steele (Savolainen A, et al., 1995; 24(4):209-11), inflammatory disease of the large intestine (IBD) (Sawai T, et al., Pediatr Surg. Int. 2001 May; 17(4):269-74) and kidney cancer (Noguchi S, et al., Buiochem Biophys. Res. Commun. 1992 Jan 31; 182(2):544-50), which further confirms the suitability of analogs of oxidized LDL for the treatment or prevention of the above diseases or disorders.

In accordance with the method of the present invention, the oxidized lipids can enter needy subject in different ways, which include, for example, oral and rectal administration, the introduction through the mucous membrane, especially in the nose, intestinal or parenteral administration, including intramuscular, subcutaneous and intramedullary injections as well as intrathecal, direct intraventricular, intravenous, intraperitoneal, nasal, or intraocular injections. However, as described in detail in this description of the invention and further illustrated in the Examples section, the preferred methods introduction the deposits are oral, through the mucous membrane, nasal, intradermal (subcutaneous and intraperitoneal routes of administration.

In one embodiment of the invention intraperitoneally injected with 0.1-100 mg/kg of oxidized lipid in an acceptable carrier, which includes, but is not limited to, PBS or glycerol, one to three times per week in accordance with the scheme of constant or alternating injection.

In another embodiment of the invention in the nose impose 0.1-100 mg/kg of oxidized lipid in an acceptable carrier, which includes, but is not limited to, PBS or glycerol, one to three times per week in accordance with the scheme of constant or alternating injection.

In another embodiment of the invention subcutaneously injected with 0.1-100 mg/kg of oxidized lipid in an acceptable carrier, which includes, but is not limited to, PBS or glycerol, one to three times per week in accordance with the scheme of constant or alternating injection.

In another embodiment, the invention is orally administered at 0.1-100 mg/kg of oxidized lipid in an acceptable carrier, which includes, but is not limited to, PBS or glycerol, one to three times per week in accordance with the scheme of constant or alternating injection.

The method according to the present invention may further include inserting the above compositions together with one or more additional compounds, rednaznachenie for the treatment or prevention of the above inflammation, associated with endogenous oxidized lipid.

The method according to the present invention may include joint introduction carried out prior to, simultaneously with or after the introduction of oxidized lipids, a therapeutically effective amount of one or more additional compounds, with the pharmaceutical composition of the present invention may contain in addition to the compounds of the present invention such additional connections.

Typical examples of additional compounds, which are intended for the treatment or prophylaxis of inflammation associated with the above endogenous oxidized lipid, and therefore suitable for use in this embodiment of the present invention, include, but are not limited to, inhibitors of HMGCoA-reductase (statins), mucosal adjuvants, corticosteroids, tools steroid anti-inflammatory therapy, means of non-steroidal anti-inflammatory therapy, analgesics, growth factors, toxins, inhibitors of the protein, providing transfer complex cholesterolemia ether (CETP), agonists proliferating activated receptor peroxisomes (PPAR), protivoerozionnye tools, antiproliferative tools, ezetimib, nicotinic acid inhibitors of squalene, ApoE Milano, HSP, beta-2-glycoprotein I and any derivative and Ana the exhaust gas.

Inhibitors of HMGCoA-reductase (statins) are well-known drugs that effectively reduce the levels of LDL-cholesterol in the inhibition of the enzyme that regulates the rate of production of cholesterol, and increased excretion by the liver LDL-cholesterol present in the blood. Non-limiting examples of commonly prescribed statins include atorvastatin, fluvastatin, lovastatin, pravastatin and simvastatin.

Ezetimibe is the first agent in a new class of inhibitors of cholesterol absorption, which effectively and selectively inhibits pogashenie dietary and biliary cholesterol at the brush borders of intestinal epithelium, without affecting the absorption of triglycerides or fat-soluble vitamins. Thus, ezetimibe reduces overall intake of cholesterol in the liver and induces increased expression of LDL receptors, which contributes more to remove LDL-C from plasma.

Peroxisome is odnomikronnoi organelle that is present in almost all eukaryotic cells. One of the most important metabolic processes mediated by peroxisome is β-oxidation of fatty acids with long and very long chain. Peroxisome also involved in the synthesis of bile acids, cholesterol and plasmalogen, the metabolism of amino acids and purine.

Nicotinic acid is one is known by the agent, reducing the levels of total cholesterol, LDL-cholesterol and triglycerides and increases the levels of HDL-cholesterol. There are three types of medicines on the basis of nicotinic acid: means of immediate action, delayed action and prolonged action. Nicotinic acid or Niacin, is a water-soluble vitamin, improves all lipoproteins when the doses in excess of the need for this vitamin.

Squalene, an isoprenoid compound that is structurally similar to beta-carotene, is promezhutochnym metabolite in the synthesis of cholesterol. The human body absorbs about 60 percent of dietary squalene. He moved in serum, being associated with VLDL, and distributed in all human tissues, with maximum concentrations in the skin, where it is one of the main components of the surface lipids of the skin. Inhibitors of squalene (e.g., monooxygenase and synthase) serve as inhibitors of the biosynthesis of cholesterol.

Agonists proliferating activated receptor (PPAR)such as fibrates, are members of the superfamily of nuclear receptors, activated fatty acids, which play an important role in the metabolism of lipids and glucose and are used for the treatment of metabolic diseases associated with obesity such as hyperlipemia, resistance to insulin and coronary heart disease. Fibrates are usually effectively reduce elevated levels of triglycerides and cholesterol in plasma and act as agonists of PPAR. The most pronounced effect of fibrates is to reduce the concentration of lipoproteins with high triglycerides (TRL) in the plasma. Levels of LDL-cholesterol (LDL-C) is usually reduced in subjects with elevated baseline concentrations in plasma, and the levels of HDL-cholesterol (HDL-C) are usually raised at low initial concentrations in the plasma. Non-limiting examples of commonly prescribed fibrates include bezafibrat, gemfibrozil and fenofibrate.

Inhibitors of the protein, providing transfer complex cholesterolemia ether (CETP) plays an important role in atherogenesis by reducing the accumulation of complex cholesterolemia ester in macrophages and arterial walls, resulting in reduced education xantana cells and the absorption of cholesterol. The most promising currently known inhibitor SETR is avasimibe.

ApoA-I Milano, commonly used in recombinant form complex with phospholipid (etc-216), causes significant regression of atherosclerosis of the coronary arteries.

Found that co-administration of adjuvants mucosa is important to prevent the invasion of infectious agents through p. the surface of the mucous membranes. In the initial stages of induction of the immune response of mucosal absorption of oral or nasal insertion of antigens is achieved thanks to the unique combination antigen-discharging cells, M-cells located in folliculosebaceous epithelium (FAE) inductive sites. After the successful absorption of antigens are processed immediately and are represented underlying dendritic cells (DC).

Non-limiting examples of means nonsteroidal anti-inflammatory therapies include oxicam, such as piroxicam, isoxicam, tenoxicam, sudoxicam and CF-14304; the salicylates, such as aspirin, disalcid, benorilate, trilisate, safarin, solprin, diflunisal and fendosal; derivatives of acetic acid, such as diclofenac, fenclofenac, indomethacin, sulindac, tolmetin, isoxepac, furofenac, tiopinac, zidometacin, acemetacin, fentiazac, zomepirac, clidanac, oxepin, felbinac and Ketorolac; fenamate, such as marennikova, meclofenamate, flutamida, niflumova and taftalenovaya acid; derivatives of propionic acid, such as ibuprofen, naproxen, benoxaprofen, flurbiprofen, Ketoprofen, fenoprofen, fenbufen, indoprofen, pirprofen, carprofen, oxaprozin, pranoprofen, miroprofen, tioxaprofen, suprofen, alminoprofen and tiaprofenic acid; pyrazoles such as phenylbutazone, EXIF button, feprazone, azapropazone and trimeton.

Non-limiting examples of means steroidal anti-inflammatory therapy include, but are not limited to, corticosteroids such as hydrocortisone, hydroxytriazine, alpha-methyl dexamethasone, dexamethasone phosphate, dipropionate beclomethasone, valerate clobetasol, desonide, desoximetasone, acetate hypertension, dexamethasone, dichloride, diacetate diflorasone, valerate of diflucortolone, Flatiron, the acetonide of fluchloralin, fludrocortisone, pialat flumetazon, acetonide fluocinolone, fluocinonide, complicated butyl esters fluorine, fluocortolone, acetate of fluprednidene (fluprednidene), flurandrenolide, halcinonide, hydrocortisone acetate, butyrate hydrocortisone, methylprednisolone, triamcinolone acetonide, cortisone, ortodoxo, Platonic fludrocortisone, diacetate diflorasone, flurandrenolide, fludrocortisone, diacetate diflorasone, acetonide flurandrenolide, Madison, antenatal, aminated, betamethasone and the balance of its esters, chloroprednisone, acetate chloroprednisone, clocortolone, clestine, dichloride, difluprednate, fluchloralin, flunisolide, formation, flaperon, fluprednisolone, hydrocortisone valerate, cyclopentylpropionate hydrocortisone, hydrocortamate, meprednisone, paramethasone, prednisolone, prednisone, beclomethasone dipropionate,triamcinolone, and mixtures thereof.

Non-limiting examples of analgesics (painkillers) include aspirin and other salicylates (such as choline salicylate and magnesium), ibuprofen, Ketoprofen, naproxen sodium, and acetaminophen.

The growth factors are hormones, fulfilling a number of functions, including regulation of the formation of molecules intercellular adhesion, the change in cell proliferation, increased vascularization, increased collagen synthesis, regulation of bone metabolism and modify cell migration in this area. Non-limiting examples of growth factors include insulin-like growth factor 1 (IGF-1), transformylase β-growth factor (TGF-β), bone morphogenic protein (BMP) and the like.

Non-limiting examples of toxins include choleragen, which also doubles as an adjuvant.

Non-limiting examples of antiproliferative funds include alkilirutmi agent, such as nitrogen mustard, ethylenimine and methylmelamine, alkylsulfonate, nitrosoanatabine and threaten; an antimetabolite, such as an analogue of folic acid, an analogue of pyrimidine and purine analog; natural product such as a Vinca alkaloid, epipodophyllotoxin, antibiotic, enzyme, Texan and modifier biological reactions; various agents, such as coordination complex of platinum, anthracenedione, anthracyclin, substituted urea, production of the annual methylhydrazine or adrenal cortical suppressor; hormone or an antagonist such as adrenocorticosteroid, a progestin, an estrogen, an antiestrogen, an androgen, an antiandrogen or an analogue of gonadotropin-releasing hormone. Specific examples of chemotherapeutic agents include, for example, nitrogen mustard, epipodophyllotoxin, an antibiotic, a coordination complex of platinum, bleomycin, doxorubicin, paclitaxel, etoposide, cyclophosphamide 4-HE and cisplatin.

Samasta HSP consists of approximately 25 proteins of different molecular weight with highly conserved structures. Almost all people have cellular and humoral immune responses against microbial hitokage protein 60 (HSP60). Because of the high degree of antigenic homology existing between microbial (bacterial or parasitic) and human HSP60, immunity against microbes risk of cross-reactivity with human HSP60 expressed by endothelial cells of arteries under stress. Valid autoimmunity against the modified autologous HSP60 can also run this process (Wick et al., Atherosclerosis as an autoimmune disease: an update. TRENDS in Immunology. 2001; 22(12):665-669). HSP participates as autoantigen target in several experimental autoimmune diseases (acrit, diabetes type I). Antibodies against HSP65 and against HSP60 were clearly associated with atheromatous lesions in humans. Research done is by using rabbits and mice, show that the occurrence of HSP65-induced immune response as a result of immunization with the recombinant protein or mixture of Mycobacterium tuberculosis with a high content of HSP65 enhances atherogenesis. Using autoimmune processes against HSP65 as a possible antigen candidate, which causes a state of immunological tolerance in the “tolerization” mucous membrane, to block these reactions, the authors of the present invention found the weakening of the initial stage of atherosclerosis in mice that oral was administered HSP65, compared with mice that oral was administered BSA or PBS (Harats et al., Oral tolerance with heat shock protein 65 attenuates mycobacterium tuberculosis dosage and high fat diet driven atherosclerosis lesions. J. Am. Coll. Cardiol. 2002; 40:1333-1338); this result was confirmed by Maron, which showed that nasal vaccination with HSP reduces the inflammatory process associated with atherosclerosis (Maron et al., Mucosal administration of heat shock protein-65 decreases atherosclerosis and inflammation in aortic arch of low density lipoprotein receptor-deficient mice. Circulation. 2002; 106:1708-1715).

Beta-2-glycoprotein I (2GPI) is fosfolipidozawisimah protein that serves as a target for prothrombotic antibodies against phospholipid. Recently it was found that this protein causes immunopositive reaction and enhances atherosclerosis in mice. β-Antibodies to beta-2-GPI possess the ability is activated monocytes and endothelial cells and can induce immune responses 2GPI in mice with an increased risk of atherosclerosis, accelerating the development of this disease. When 2GPI-reactive lymph node and spleen cells transferred into mice with absence of the LDL receptor, they stimulated the formation of fatty streaks, confirming direct proatherogenic role 2GPI-specific lymphocytes. Induction of immunological tolerance to 2GPI in the preliminary oral administration of antigen significantly reduce the degree of atherosclerotic lesions. Thus, 2GPI is a putative modulator of atherosclerotic plaques, and therefore can probably be used as immunomodulator development of plaques. Oral administration 2GPI inhibits the reactivity of the cells of the lymph node in relation 2GPI in mice immunized against human protein. In cells of the lymph node 2GPI-tolerant mice immunized against 2GPI, increased production of IL-4 and IL-10 at the initiation of the corresponding protein. Thus, oral administration of 2GPI is effective suppression of atherogenesis in mice (George et al. Suppression of early atherosclerosis in LDL-receptor deficient mice by oral tolerance with beta2-glycoprotein I. Cardiovasc. Res. 2004; 62(3):603-9).

Additional objectives, advantages and distinctive features of the present invention will be obvious to specialists in this field the tee when reading the following examples, do not limit the invention. In addition, all variants of implementation and the objects of the present invention, discussed in the description of the invention and expressed in the following claims, find experimental confirmation in the following examples.

Examples

The following examples, which, together with the description of the invention illustrate the invention without limiting its scope.

The nomenclature used in the description of the invention and laboratory procedures, refers to the biochemical and immunological methods. Such methods are extensively described in the scientific literature. See, for example, the publication "Cell Biology: A Laboratory Handbook", Volumes I-III Cellis, J. E., ed. (1994); "Current Protocols in Immunology" Volumes I-III Coligan J. E., ed. (1994); Stites et al. (eds), "Basic and Clinical Immunology" (8th Edition), Appleton&Lange, Norwalk, CT (1994); Mishell and Shiigi (eds), "Selected Methods in Cellular Immunology", W. H. Freeman and Co., New York (1980); existing immunoassays are also fully described in the patent and scientific literature; see, for example, U.S. patents№№ 3791932, 3839153, 3850752, 3850578, 3853987, 3867517, 3879262, 3901654, 3935074, 3984533, 3996345, 4034074, 4098876, 4879219, 5011771 and 5281521 and publication “Methods in Enzymology” Vol. 1-317, Academic Press; Marshak et al., which are fully incorporated into this description by reference. In the description of the invention also includes other General references. It is believed that these methods are well known in this field and represent who go only for the convenience of the reader. All information contained in the publications included in this description by reference.

Materials and General methods

Animals

Mice with absence of APO-E. Mouse in absence of the apolipoprotein E (APO-E KO)used in these experiments, are prone to atherosclerosis in C57BL/6J-Apoetmlunc. In mice, homozygous in respect of Apoe mutationstmluncthere is a considerable increase in levels of total cholesterol in plasma, which affects neither age nor sex. Fat strips in the proximal aorta was detected at the age of 3 months. Lesions increase with age and progress with the formation of lesions that contain less lipid, but more elongated cells, typical of a later stage prematureclaritin defeat.

Create a line. Mutant line Apoetmluncwas created in the laboratory of Dr. Nobuyo Maeda at the University of North Carolina at chapel hill. Used cell line E14Tg2a ES isolated from 129 mice. Used plasmid was named pNMC109, and the original line was designated T-89. Line C57BL/6J strain was obtained from ten back-crossing mice with Apoe mutationtmluncmice C57BL/6J (Plump et al., Severe hypercholesterolemia and atherosclerosis in apolipoprotein-E deficient mice created by homologous recombination in ES cells. Cell 1992; 71: 343-353; Zhang et al. Spontaneous hypercholesterolemiaand arterial lesions in mice lacking apolipoprotein E. Science 1992; 258: 468-471).

Mice were kept in hospital for animals Sheba (Tel Hashomer, Israel) with a 12-hour cycle of alternation of light and dark at 22-24°C and were fed laboratory food with normal fat content (laboratory food to Purina rodent No. 5001), including 0,027% cholesterol, and about 4.5% of the total fat, water was given on demand.

Mouse LDL-RD. Mouse LDL-RD [LDLr<mlHer>LDL-/-(C57B/6 50% JSL 25% I129 25%)] at the age of 8-12 weeks were obtained from hospital for animals Hadassah (Hadassah Hospital, Israel).

Rats Lewis. Male Lewis rats aged 9-11 weeks were provided by Harlan laboratories (Israel).

Immunization

I. Intraperitoneal immunization ALLE. Similar simple ether phospholipid (ALLE D+L) was combined with purified derivative protein of tuberculin (PPD). The original solution ALLE (D+L) was dissolved in ethanol (99 mg/ml). 5 mg ALLE (D+L) (a 50.5 μl of the original solution) was diluted to 5 mg/ml of 0.25 m solution of phosphate buffer, pH 7,2, stirring at 0°C. (bath with ice). 1.5 mg of D - and L-ALLE in 300 μl of phosphate buffer was added to 0.6 mg PPD dissolved in 300 μl of phosphate buffer. Hydrochloride of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (5 mg; Sigma, St Louis. MO), dissolved in 50 μl of water was added with stirring for 20 minutes at 4°C. the Remaining active sites were blocked with 100 μl of 1 M solution of glycine. United compounds were dialyzed against saline phosphate buffer(PBS), brought up to 3 ml with PBS, and stored at 4°C. Immunization 0.3 ml (150 µg) of antigen per mouse were performed intraperitoneally 4 times in 2 weeks.

II. Subcutaneous immunization with oxidized LDL person. Oxidized LDL person has been collected from human plasma (density 1,019-1,063 g/ml) ultracentrifugation and oxidized Cu during the night (which was added 15 μl of 1 mm CuSO4each ml of LDL, pre-diluted to a concentration of 1 mg/ml). Oxidized LDL were dialyzed against PBS and filtered. For immunization with oxidized LDL was dissolved in PBS and mixed with an equal volume of incomplete adjuvant's adjuvant. Immunization was performed as a single subcutaneous injection of 50 μg of antigen/mouse in 0.2 ml volume. Mice were immunized once in one to three days after the last oral administration and squashed 7-10 days after immunization.

Determination of cholesterol. After the end of the experiment using cardiac puncture received 1-1,5 ml of blood, to each sample was added to 1000 units/ml heparin, and determined the levels of total cholesterol in plasma automated enzymatic method (Boehringer Mannheim, Germany).

The analysis method FPLC. Analysis of cholesterol and lipids in lipoproteins by liquid chromatography protein a quick resolution performed in the system for FPLC (Pharmacia LKB FRAC-200, Pharmacia, Peapack, NJ) with a column filled with supersoil 6 HR 10/30 (Amersham Pharmacia Biotech, Inc. Peapack, NJ). In the sampling tube automatic sampler required minimum sample volume of 300 ál (blood collected from 3 mice was diluted in ratio 1:2 and filtered before loading) to fill 200 ál loop for the sample. Collected fractions 10-40, each of which consisted of 0.5 ml 250 ál sample from each fraction was mixed with freshly prepared cholesterol or triglyceride reagent, incubated for 5 minutes at 37°C and analyzed by the spectrophotometric method at 500 nm.

Assessment of atherosclerosis. Quantification of atherosclerotic lesions in the form of fatty strips produced by calculating the size of the lesions in the aortic sinus in accordance with the description given in the scientific literature (Paigen et al., Quantitative assessment of atherosclerotic lesions in mice. Atherosclerosis 1987; 68:231-140), and calculate the value of the lesions in the aorta. After perfusion Tris-buffer with EDTA in saline animals were removed the heart and the aorta and gently scraped peripheral fat. The upper section of the heart was immersed in the environment of the EAST (10,24 wt./wt.% polyvinyl alcohol; 4.26 deaths wt./wt.% polyethylene glycol; 85,50 wt./wt.% inert ingredients) and froze. Analyzed every second slice (thickness 10 μm) sinus of the aorta (400 μm). The distal part of the aortic sinus were identified on the three doors of the valve, which are the Estai connect the aorta to the heart. Sections were evaluated in relation to deposits of fat strips after staining with oil red O. the Area of damage on a single slice was evaluated on the grid (Rubin et al. Inhibition of early atherogenesis in transgenic mice by human apolipoprotein A-I. the Nature 1991; 353:265-267) by counting unidentified numbered samples performed by the observer. The aorta is dissected from the heart and removed the surrounding adventitia tissue. The aorta were fixed and stained vessels Sudan in accordance with the description given in the scientific literature (Bauman and Mangold, J. Org. Chem. 31: 498, 1966).

Measurement and quantification of atherosclerotic lesions in the plasma. The levels of total cholesterol and total triglycerides in plasma were measured by the method COBAS MIRA. The dead animals were removed hearts and crisisi aortic root were stained with oil red O. the Area of atherosclerotic lesions was determined by computer analysis (Image Pro Plus) and the results were confirmed by examination under a microscope.

Analyses of cell proliferation. Mice orally introduced ALLE, POVPC or PBS similar to the description of the method of assessment of atherosclerosis and were immunized through one day after the last oral administration of oxidized LDL obtained from the above purified LDL person.

Proliferation was determined eight days after immunization with oxidized LDL as follows: spleen or limfaticheskie nodes was obtained by grinding the tissue on a sieve with a mesh size of 100 mesh. (lymph nodes with completed immunization and spleen without immunization). Erythrocytes were literally cold sterile double-distilled water (6 ml) for 30 seconds and add 2 ml of 3.5% NaCl. Added incomplete medium (10 ml), cells were centrifuged for 7 minutes at a speed of 1700 rpm, re-suspended in RPMI medium and proizvoditi counting in hemocytometer when diluted in the ratio of 1:20 (10 μl cells + 190 ál Trypanosoma blue). Proliferation was measured by introducing a [3H]-thymidine into the DNA of three identical samples of 100 µl of erythrocyte mass (2,5×106cells/ml) in 96-well titration the microplate. Added three of the same sample oxidized LDL (0-10 μg/ml, 100 µl/well), cells were incubated for 72 hours (37°C, 5% CO2and humidity of about 98%) was added 10 μl of3[H]-thymidine (0.5 to MX/well). Cells were incubated another day, then harvested, transferred to glass fiber filters using a harvester (Brandel) and made calculations in a β-counter (Lumitron). For analysis of cytokines collected supernatant without adding3[H]-thymidine and were analyzed using ELISA method.

A separate group of mice orally introduced ALLE or PBS and were immunized oxidized LDL by the method described above through one day after oral administration of the last dose. Lymph nodes of inguinal region (obtained after 8 days after immunization) were removed from 3 mice in each group for the study of cell proliferation. 1×106cells per ml were incubated in the form of three identical samples within 72 hours in 0.2 ml of culture medium in the cell tiralongo microplate in the presence of 10 μg/ml oxidized LDL. Proliferation was measured by introducing a [3H]-thymidine into DNA during the last 12 hours of incubation. The results were expressed as stimulation index (S.I): the ratio of the average radioactivity (counts per minute) of the antigen to the average background radioactivity (counts per minute)obtained in the absence of antigen. The standard deviation was always equal to <10% of the average number of pulses per minute.

Evaluation of inflammatory markers in serum. Serum was separated by centrifugation and stored at -70°C. Analysis of inflammatory markers was performed by the method of ELISA (IL-10; R&D and SAA; BIOSOURCE).

Analysis by the method of RT-PCR. Aorta, spleen and small intestine were removed from immunized and non-immunized mice (under sterile conditions) and frozen in liquid nitrogen. Bodies were crushed on the sieve and received RNA using the Rneasy kit (QIAGEN). Arazzi RNA was investigated in the spectrophotometer and normalized relative to β-actin. Reverse transcription of RNA into cDNA and polymerase reaction synthesis chain (PCR) using the seed was performed using the kit to perform RT-PCR in one tube Titan (ROCHE). The results were obtained on 1% agar is testing the gel and documented on film.

The statistical analysis. The ANOVA analysis one variable was used to compare independent values. As a statistically significant value was adopted value of p<0,05.

Example I

Synthesis of immunomodulatory antigen presenting a simple ester of 2,5'-allegedlly (ALLE and POVPC

ALLE

1-hexadecyl-2-(5'-oxapentane)-sn-glycero-3-phosphocholine

POVPC

1-hexadecanoyl-2-(5'-oxovalerate)-sn-3-phosphocholine

Synthesis of a simple ester of 2,5'-allegedlly (ALLE)

A simple ester of 2,5'-allegedlly (ALLE) was synthesized in accordance with the modified General methods of synthesis of analogues of simple ether of lecithins, described in the publications of H. Eibl, et al., Ann. Chem. 709:226-230(1967), W.J. Baumann and H.K. Mangold, J. Org. Chem. 31498 (1996), E. Baer and Buchnea JBC. 230447 (1958), G. Halperin et al., Methods in Enzymology 129838-846 (1986). Following the method relates to the production of compounds, two-dimensional image which is presented in figure 1.

Simple ether hexadecylamine. D-Acetoglyceride (4 g) for the synthesis of L-ALLE or L-acetoglyceride for the synthesis of D-ALLE, powdered potassium hydroxide (about 10 g) and hexadecylamine (9.3 g) in benzene (100 ml) primatively and was heated under reflux for 5 hours, removing the water formed by azeotropic distillation (compare publication W.J. Baumann ad H.K. Mangold, J. Org. Chem. 29:3055, 1964 and F. Paltauf, Monatsh. 99:1277, 1968). The volume of solvent was gradually reduced to about 20 ml, the mixture was cooled to room temperature and was dissolved in simple ether (100 ml). The resulting solution was washed with water (2×50 ml) and solvent was removed under reduced pressure. To the residue was added 100 ml of a mixture methanol:water:concentrated hydrochloric acid (90:10:5), and the mixture was heated under reflux for 10 minutes. The product was extracted with simple ether (200 ml) and then washed with water (50 ml), 10% sodium hydroxide solution (20 ml) and again water (20 ml volumes) to a neutral state. The solvent was removed under reduced pressure and the product (8.8 g) was led from hexane, thus obtaining of 7.4 g of pure 1-hexadecyl-glicerynowego ester (compound I, figure 1) for the synthesis of D-ALLE or 3-hexadecylpyridinium ether for synthesis of L-ALLE.

5-Hexanicotinate. A mixture of 5-HEXEN-1-ol (12 ml) and dry pyridine (25 ml) was cooled to a temperature of -4°C to -10°C in an ice bath salt. Was added dropwise methanesulfonanilide (10 ml) for 60 minutes and kept the mixture at 4°C for 48 hours. Added ice (20 g), and the mixture was left to stand for 30 minutes and the product was extracted by a simple ether (200 ml). The organic phase is washed with water (20 ml), 10% hydrochloric acid, 10% of concrete is sodium bicarbonate (20 ml) and again water (20 ml). The solvent is evaporated and the crude product was chromatographically on silica gel 60 (100 g)using a mixture of CHCl3:EtOAc (80:20) as eluent, resulting in 14 g of 5-hexanitrocobaltate.

1 Hexadecylamine-3-trityloxy-2-propanol (for D-ALLE) or 3-hexadecylamine-1 trityloxy-2-propanol (for L-ALLE) (compound II). 1-Hexadecyloxypropyl (for D-ALLE) or 3-hexadecyloxypropyl (for L-ALLE) (7.9 g), triphenylarsine (8,4 g) and dry pyridine (40 ml) was heated at 100°C for 12 hours. The mixture was cooled, was added 300 ml of a simple ether and 150 ml of cooled with ice water and the reaction mixture was transferred into a separating funnel. The organic phase is then washed with 50 ml ice water, 1% potassium carbonate solution (to the ground state) and 50 ml of water, then dried over anhydrous sodium sulfate. The solvent is evaporated, the residue was dissolved in 150 ml of warm petroleum ether and the resulting solution was cooled at 4°C over night. The precipitate was filtered, the filtrate was evaporated and the residue precrystallization from 20 ml of ethyl acetate at -30°C, while receiving of 8.2 g of 1-hexadecylamine-3-trityloxy-2-propanol (for D-ALLE) or 3-hexadecylamine-1 trityloxy-2-propanol (for L-ALLE) (compound II, figure 1), melting point 49°C.

1-Hexadecyl-2-(5'-hexenyl)gliterry ether (for D-ALLE) or 3-hexadecyl-2-(5'-hexenyl)gliterry ether(for L-ALLE) (compound IV). 1 Hexadecylamine-3-trityloxy-2-propanol (for D-ALLE) or 3-hexadecylamine-1 trityloxy-2-propanol (for L-ALLE) (compound II, figure 1) (5.5 g) was atrificial 5-hexanicotinate using powdered potassium hydroxide in benzene solution as above described. The crude product 1-hexadecylamine-2-(5'-hexenoate)-sn-3-tritylation (for D-ALLE) or 3-hexadecylamine-2-(5'-hexenoate)-sn-3-tritylation (for L-ALLE) (compound III, figure 1) was dissolved in 100 ml of a mixture methanol:water:concentrated hydrochloric acid (90:10:5) and the reaction mixture was heated under reflux for 6 hours. The product was extracted with simple ether, washed with water and the solvent was removed. The residue was dissolved in petroleum ether (100 ml) and kept at 4°C during the night, resulting in the deposition occurred mostly of triphenylcarbinol. The product was filtered and solvent was removed from the filtrate, after which the crude product was chromatographically on silica gel 60 (40 g)using a mixture of chloroform:petroleum ether (1:1) as eluent, resulting in 1.8 g of pure 1-hexadecyl-2-(5'-hexenyl)glicerynowego ether (for D-ALLE) or 3-hexadecyl-2-(5'-hexenyl)glicerynowego ether (for L-ALLE) (compound IV, figure 1).

1-Hexadecyl-2-(5'-hexenyl)-sn-glycero-3-phosphocholine (for D-ALLE) or 3-hexadecyl--(5'-hexenyl)-sn-glycero-1-phosphocholine (for L-ALLE) (compound V). The following method is a modification of the method described in the publication of H. Eibl, et al. Ann. Chem 709:226-230, 1967.

A solution of 1-hexadecyl-2-extirpative ether (for D-ALLE) or 3-hexadecyl-2-extirpative ether (for L-ALLE) (compound IV, figure 1) (2 g) in dry chloroform (15 ml) was added dropwise to a stirred cooled solution (from -4°C to -10°C) dry triethylamine (3 ml) and 2-bromatological (1.25 ml, obtained as described below) in dry chloroform (15 ml) for 1 hour. The mixture was stirred at room temperature for 6 hours and then was heated up to 40°C for 12 hours. The obtained dark brown solution was cooled to 0°C was added a 0.1 M solution of potassium chloride (15 ml). The mixture was left to warm to room temperature and was stirred for 60 minutes. Was added methanol (25 ml) and chloroform (50 ml), the organic phase was washed with 0.1 M hydrochloric acid (20 ml) and water (20 ml). The solvent is evaporated, the crude product was dissolved in methanol (15 ml), the solution was transferred into a test tube for culture and cooled in an ice bath salt. Added cold trimethylamine (3 ml, -20°C) and tightly corked test tube. The mixture was heated to 55°C for 12 hours, cooled to room temperature and the solvent evaporated in a stream of nitrogen. The residue was extracted with a mixture of chloroform:meta is ol (2:1) (25 ml) and washed with 1 M potassium carbonate solution (10 ml) and water (2×10 ml). The solvent was removed under reduced pressure and the crude products were chromatographically on silica gel 60 (20 g)using a mixture of chloroform:methanol (60:40), the result of which was obtained 1.5 g of 1-hexadecyl-2-(5'-hexenyl)-sn-glycero-3-phosphocholine (for D-ALLE) or 3-hexadecyl-2-(5'-hexenyl)-sn-glycero-1-phosphocholine (for L-ALLE) (compound V, figure 1). The structure of compound V was confirmed by NMR and mass spectrometry.

1-Hexadecyl-2-(5'-oxapentane)-sn-glycero-3-phosphocholine (for D-ALLE) or 3-hexadecyl-2-(5'-oxapentane)-sn-glycero-1-phosphocholine (for L-ALLE) (compound VI). A solution of compound V (0.5 g) in formic acid (15 ml) and 30% hydrogen peroxide (3.5 ml) was stirred at room temperature overnight. The reaction mixture was diluted with water (50 ml) and was extracted with a mixture of chloroform:methanol (2:1) (5×50 ml). The solvent is evaporated under reduced pressure, the residue was mixed with methanol (10 ml) and water (4 ml) and stirred at room temperature for 60 minutes. Then added 80% phosphoric acid (2 ml) and metaperiodate potassium (0.8 g). The mixture was stirred at room temperature overnight, diluted with water (50 ml) and was extracted with a mixture of chloroform:methanol (2:1) (50 ml). The organic phase was washed with a 10% solution of sodium bisulfite (10 ml) and water (10 ml). The solvent was removed under reduced pressure, the crude product was chromatographically on when likehere 60 (10 g), using a mixture of chloroform:methanol (1:1) as eluent, resulting in 249 mg 1-hexadecyl-2-(5'-oxapentane)-sn-glycero-3-phosphocholine (for D-ALLE) or 3-hexadecyl-2-(5'-oxapentane)-sn-glycero-1-phosphocholine (for L-ALLE) (compound VI, figure 1)with a value of Rf0,15 (system for TLC, chloroform:methanol:water (60:40:8)) and is characterized by a positive reaction with dinitrophenylhydrazine. The chemical structure of compound VI was confirmed by NMR and mass spectrometry.

In accordance with an alternative method ethylene group, was turned into an aldehyde group by ozonation and catalytic hydrogenation of calcium carbonate on palladium.

Getting 2-bromatological. 2-Bromatological was obtained by adding dropwise have just surpassed 2-bromoethanol (0.5 M, obtained by the method described in the publication Gilman Org. Synth. 12:117, 1926) to a cooled with ice to a solution that only distilled phosphorus oxychloride (0.5 M) in dry chloroform for 1 hour, then the reaction mixture was heated under reflux for 5 hours and subjected to vacuum distillation (boiling point : 66-68°C under a pressure of 0.4-0.5 mm Hg). The reagent was stored until use (-20°C) in nitrogen atmosphere in a small sealed ampoules (W.H. Hansen et al. Lipids 17(6):453-459, 1982).

Synthesis of 1-hexadecyl-2-(5'-carboxybutyl)-sn-glycero-3-fo is tholine (CI-201). 1-Hexadecyl-2-(5'-oxapentane)-sn-glycero-3-phosphocholine (compound VI obtained in the above way), 0.55 g (0.001 mol), was dissolved in t-BuOH (30 ml). Was added dropwise a solution of NaClO2(0.9 g, 0.01 mol) and NaH2PO4(0.96 g, 0.07 mol) in 25 ml of water for 30 minutes and the mixture was stirred at room temperature for 3 hours. The reaction mixture was acidified to pH=3 with concentrated hydrochloric acid and was extracted with a mixture of chloroform:methanol (2:1). The organic phase was separated and the solvent evaporated. The residue was purified by chromatography on silica gel using a mixture of chloroform:methanol:water (70:27:23), the result of which was obtained 1-hexadecyl-2-(5'-carboxybutyl)-sn-glycero-3-phosphocholine (0,42 g, yield 72%). Chemical structure (compound VII, figure 10) was confirmed by NMR and mass spectrometry.

Synthesis of 1-hexadecyl-2-(5',5'-dimethoxyphenoxy)-sn-glycero-3-phosphocholine. 1-Hexadecyl-2-(5'-oxapentane)-sn-glycero-3-phosphocholine (compound VI obtained in the above way), 0.50 g (0.89 mmol), was dissolved in formic acid (15 ml) was added 30% hydrogen peroxide (3.5 ml). The reaction mixture was stirred over night at room temperature. Was added water (50 ml) and product was extracted with a mixture of chloroform:methanol (2:1) (2×50 ml). The organic phase is washed with 10% aqueous sodium bicarbonate solution (10 ml) and water (10 ml) and RAS shall foretell was removed under reduced pressure. The residue (0.4 g) was dissolved in methanol (10 ml), was added 10% aqueous sodium hydroxide solution (4 ml) and the reaction mixture was stirred at room temperature for 1 hour. Then added 80% phosphoric acid (2 ml) and metaperiodate potassium (0.8 g) and continued stirring the reaction mixture overnight. Was added a mixture of chloroform:methanol (2:1) (50 ml), the organic phase is washed with 10% aqueous solution of sodium bisulfite (10 ml) and water (10 ml) and the solvent was removed in vacuum. The residue (0.3 g) was purified by chromatography on silica gel (10 g)using a mixture of chloroform:methanol (60:40-40:60) as the gradient eluent, the result of which was obtained 1-hexadecyl-2-(5',5'-dimethoxyphenoxy)-sn-glycero-3-phosphocholine (0.25 g, yield 46%). Chemical structure (compound VIIIa, figure 10) was confirmed by NMR and mass spectrometry.

Synthesis of 1-hexadecyl-2-(5',5'-deoxypentose)-sn-glycero-3-phosphocholine. The crude 1-hexadecyl-2-(5'-oxapentane)-sn-glycero-3-phosphocholine (compound VI obtained as described above), 50 mg (0,088 mmol), was dissolved in ethanol (10 ml) under nitrogen atmosphere. Added triethylorthoformate (0,053 ml, 0,0476 g, 0.32 mmol) and 3 drops of concentrated sulfuric acid and the reaction mixture was stirred over night at room temperature. Then added dichloromethane (75 ml) and the reaction mixture was transferred into a separating funnel, after which avatele washed with water (75 ml), 2.5% aqueous sodium bicarbonate solution (75 ml) and water (75 ml) and dried over anhydrous sodium sulfate. The reaction mixture was filtered and the solvent was removed in vacuum, while receiving 50 mg of the crude 1-hexadecyl-2-(5',5'-deoxypentose)-sn-glycero-3-phosphocholine. The structure was confirmed by NMR and mass spectroscopy (compound VIIIb, figure 10).

Synthesis of 1-hexadecanoyl-2-(5'-oxovalerate)-sn-3-glycerophosphocholine (POVPC). A mixture of 1-hexadecanoyl-sn-3-glycerophosphocholine (compound I, figure 2) (3 g), 5-hexenoic acid (1.2 ml), 1,3-dicyclohexylcarbodiimide (DCC, of 4.05 g) and N,N-dimethylaminopyridine (DMP, 1.6 g) in dichloromethane (100 ml, freshly distilled from phosphorus pentoxide) was thoroughly stirred for 4 days at room temperature. Then the mixture was chromatographically on silica gel 60 (40 g) and the product, 1-hexadecanoyl-2-(5'-hexanoyl)-sn-3-glycerophosphocholine (2.8 g, compound II, figure 2), was suirable a mixture of chloroform:methanol (25:75). Eluent was dissolved in a mixture of 30% hydrogen peroxide:formic acid (4:15) and the solution was stirred over night at room temperature. Was added water (50 ml), the product was extracted with a mixture of chloroform:methanol (2:1) (100 ml) and the organic phase is washed with water. The solvent is evaporated under reduced pressure, the residue was dissolved in methanol (15 ml) and 10% ammonia solution (5 ml) and the resulting solution was stirred pikantnoi temperature for 6 hours. The crude 1-hexadecanoyl-2-(5',6'-dihydroxy)hexanoyl-sn-3-glycerophosphocholine (compound III, figure 2) (the structure was confirmed by NMR and mass spectrometry) were subjected to further interaction without purification. To the solution was added 80% phosphoric acid (3 ml) and metaperiodate sodium (1 g), the mixture was stirred at room temperature over night and was extracted with a mixture chloroform:methanol (2:1). The product was purified by chromatography on silica gel 60 (20 g)using a mixture of chloroform:methanol (25:75) as eluent. Received 850 mg 1-hexadecanoyl-2-(5'-oxovalerate)-sn-3-glycerophosphocholine (POVPC, compound IV, figure 2), which were characterized by chromatographic mobility of lecithin when performing TLC and positive interaction with dinitrophenylhydrazine. The structure was determined by NMR and mass spectrometry.

Alternative ethylene group was converted to the aldehyde by ozonation and catalytic hydrogenation of calcium carbonate on palladium.

Example II

Specific inhibition of atherogenesis in genetically predisposed mice (APO-E KO) immunization against L-ALLE + D-ALLE

The authors of the present invention found that immunization sustainable esterified synthetic ALLE, which is a component of LDL, reduces the formation of atherosclerotic plaques in prednisolone the different mice. 19 female mice Aro E/57 at the age of 5-7 weeks were divided into 3 groups. In group a (n=6) mice were immunized intraperitoneally in accordance with the description given in the section “Materials and methods”, 150 μg/mouse L-ALLE + D-ALLE once in 2 weeks (0.3 ml/mouse), repeating this procedure 4 times. In group b (n=6) mice were immunized with purified derived protein (PPD) toxin tuberculin once in 2 weeks (0.3 ml/mouse). In group C (n=7) mice were not immunized. Mice in all three groups took blood samples before immunization (time 0) and after one week after the second immunization for the detection of antibodies against ox LDL antibodies against ALLE and lipid profile. Assessment of atherosclerosis produced by the method described above in 4.5 weeks after the 4th immunization. Mice in all groups were weighed every 2 weeks throughout the experiment. All mice received normal feed, containing 4.5 wt.% fat (0.02% cholesterol), and water needs.

Table 1
Inhibition of atherogenesis in the immunization of mice APO-E KO via ALLE
GroupImmunization
L-ALLE+D-ALLE 150 µg/
mouse N=6
PPD
N=5
Control
group
without
immunize the AI N=7
The statistical-
Kie
data
Time 0Weight17,3±0,517,3±0,717,8±0,4P=0,780
Cholesterol435±47436±49413±44P=0,919
Triglyceride118±9112±10120±14P=0,865
EndWeight20,5±0,521,6±0,220,3±0,5P=0,123
Cholesterol299±18294±153044±22P=0,937
Triglyceride57±353±466±4P=0,075
The magnitude of time is possible (μm 2)101000±8276179500±13449210833±26714P=0.005
TGF-β pmol/ml12032±230813963±94412825±2340P=0,831
Note: the “mass” means the mass in grams; cholesterol” means the level of cholesterol in plasma and “triglyceride” refers to the level of triglycerides in plasma, expressed in mg/DL.

As shown in figure 3, the results in table I indicate a significant decrease in atheromatous lesions, measured in the tissues of the heart ALLE-immunized mice compared to PPD-immunized mice and unimmunized control mice. We detected no significant effect on other measured parameters, such as weight gain, levels of triglycerides or cholesterol in plasma or immunocompetent measured on the basis of the levels of the immunosuppressive cytokine TGF-β. Thus, immunization with synthetic ALLE, which is a component of oxidized LDL (the racemic mixture of D and L forms), substantially (>50%) protects genetically predisposed mice Aro-E TO the occurrence of the at acclerations defeat. Significant, but less than significant decrease in belascoaran was observed in mice immunized with PPD.

Example III

Inhibition of atherogenesis in genetically predisposed mice (APO-E KO) as a result of oral administration of L-ALLE and D-ALLE

Intraperitoneal immunization ester analogues components belascoaran resulted in effective inhibition of atherogenesis in mice APO-E KO (figure 1). We studied the possibility of oral administration of L - and D-ALLE to suppress atherogenesis. 34 male mice APO-E KO aged 8-10 weeks were divided into three groups. In group a (n=11) mice orally was administered via a stomach tube L-ALLE + D-ALLE suspended in PBS containing 5% ethanol (1 mg/mouse)every other day for 5 days. In group b (n=11) mice orally was administered 10 μg/mouse L-ALLE+D-ALLE suspended in PBS containing 5% ethanol, every other day for 5 days (0.2 ml/mouse). Mice in group C (n=12) was administered PBS (containing the same amount of ethanol that in groups a+B). In mice took blood samples to oral administration (time 0) and at the end of the experiment (end) to determine the lipid profile. Atherosclerosis was assessed in aortic sinus by the method described above in 8 weeks after the last oral administration. Mice were weighed every 2 weeks throughout the experiment. All mice received normal feed, containing 4.5 the AC.% fat (0.02% cholesterol), and water needs.

Table 2
Inhibition of atherogenesis in mice Aro-E TO the result of oral administration of L-ALLE and D-ALLE
GroupPBS
N=12
1 mg
ALLE N=11
10 µg ALLE
N=11
Statistical data
Time 0Weight20,7±0,621,5±0,821,1±0,8P=0,794
Cholesterol373±25379±23378±31R=0,983
Triglyceride1289890P=0,829
EndWeight27,3±0,427,4±0,524,1±0,8P<0,001
Cholesterol 303±17249±24321±15P=0,031
Triglyceride81±478±893±6P=0,146
The size of the lesion (μm2)176000±1373585278±11633103889±14320P<0,001
TGF-β pmol/ml14696±135213388±148918010±1373P=0.07
Note: the “mass” means the mass in grams; cholesterol” means the level of cholesterol in plasma and “triglyceride” refers to the level of triglycerides in plasma, expressed in mg/DL.

As shown in figure 4, the results are shown in table 2, ukazavaet to a significant weakening in the development of atherosclerosis, measured in tissues of mice that oral was administered a low dose (10 μg-1 mg/mouse) ALLE compared to unimmunized control mice. We detected no significant effect on other measured parameters, the e as weight gain, the levels of triglyceride or cholesterol in plasma or immunocompetent measured on the basis of the levels of the immunosuppressive cytokine TGF-β. Thus, oral administration of synthetic ALLE, allamagoosa component of oxidized LDL, significantly (>50%) protects genetically predisposed mice APO-E KO from atherosclerosis, and such protection comparable to the protection achieved with intraperitoneal immunization (see figure 1).

Example IV

Inhibition of atherogenesis in genetically predisposed mice (APO-E KO) in the induction of immunomodulation in oral and natalina the introduction of L-ALLE

Mechanisms of immunomodulation mediated mucosa, are active in the mucous membrane of the nose and intestines. Therefore, compared the effectiveness of suppression of atherogenesis in mice APO-E KO when natalina and oral vvedenim L - and D-ALLE. 34 male mice APO-E KO at the age of 7-10 weeks were divided into 3 groups. In group a (n=11) mice orally was administered L-ALLE suspended in PBS containing 5% ethanol (1 mg/mouse/0.2 ml)every other day for 5 days. In group b (n=11) mice were injected in the nose in the manner described under “Materials and methods”, 10 μg/mouse/10 µg L-ALLE suspended in PBS every other day for 3 days. Mice in group C (n=12) oral and nasal were administered PBS (containing the same amount of ethanol that in groups a+Century In mice took blood samples to oral administration (time 0) and at the end of the experiment (end) to determine the lipid profile. Atherosclerosis was assessed in aortic sinus as described above in 8 weeks after the last oral administration. Mice were weighed every 2 weeks throughout the experiment. All mice received normal feed, containing 4.5 wt.% fat (0.02% cholesterol), and water needs.

Table 3
The influence of the oral and nasal introduction of L-ALLE on rates of metabolism and atherogenesis in mice APO-E KO
GroupOral administration of ALLE
1 mg
(N=11)
Nasal introduction
ALLE
10 mcg (N=11)
Oral/ nasal introduction PBS
(N=12)
Statistical
data
Time 0Weight21,1±0,821,1±0,722,1±0,9P=0,624
Cholesterol362±27353±31351±27 P=0,952
Triglyceride144143138P=0,977
EndWeight23,3±1,124,2±0,224,0±0,5P=0,558
Cholesterol418±43328±18343±25P=0,084
Triglyceride82±774±679±5P=0,630
The size of the lesion (μm2)76944±1707282708±10986135455±12472P=0.007
Note: the “mass” means the mass in grams; cholesterol” means the level of cholesterol in plasma and “triglyceride” refers to the level of triglycerides in plasma, expressed in mg/DL.

As shown in figure 5, the results are shown in table 3, which led to an effective inhibition of atherogenesis, measured in tissues of mice that were injected in the nose low dose (10 μg/mouse) ALLE compared to neionizirovanne control mice. Nasal introduction like oral introduction had no significant effect on other measured parameters, such as weight gain, levels of triglycerides or cholesterol in the plasma. Thus, synthetic ALLE, which is a component of oxidized LDL, significantly (>50%) protects genetically predisposed mice APO-E KO against atherosclerosis as oral, and natalina introduction.

Example V

Suppression of specific immune response against oxLDL in genetically predisposed mice (APO-E KO) by oral administration of L-ALLE or POVPV

Immunomodulation induced by injection through the mucous membrane of the oxidized analogues LDL, may be mediated by suppression of specific immune responses to antigens associated with plaques. POVPC (1-hexadecanoyl-2-(5'-oxovalerate)-sn-glycerophosphocholine) is a non-terrestrial analogue of oxidized LDL, which unlike ALLE subject to destruction in the liver and intestine. Lymphocyte proliferation in response to oral administration of POVPC and more stable analogue ALLE measured in mice APO-E KO. 8 male mice APO-E KO at the age of 6 weeks were divided into 3 groups. In group a (n=2) mice as described above through VC the exploration probe was administered 1 mg/mouse L-ALLE, suspended in 0.2 ml PBS every other day for 5 days. In group b (n=3) mice as described above oral was administered 1 mg/mouse POVPC, suspended in 0.2 ml PBS every other day for 5 days. Mice in group C (n=3, control group) orally was administered 200 μl PBS every other day for 5 days. Immune reactivity stimulated by immunization with oxidized LDL person in the manner described under “Materials and methods”, the day after the last oral administration. One week after immunization mice were removed lymph nodes for analysis of proliferation. All mice received normal feed, containing 4.5 wt.% fat (0.02% cholesterol), and water needs.

Table 4
Suppression of immune response to ox-LDL person in mice APO-E KO in the preliminary oral administration of synthetic oxidized LDL (ALLE and POVPC)
The stimulation index (SI)
PBSPOVPCL-ALLEStatistical data
33,1±6,110,6±2,37,3±2,3P<0,01
N=3N=2
-68%-78%

As shown in figure 6, the results in table 4 indicate significant suppression of immune reactivity against the antigen oxidized LDL person, measured on the basis of inhibition of cell proliferation in the lymph nodes of mice APO-E KO. Lymphocytes of mice that oral was administered inhibiting atherogenesis dose (1 mg/mouse) ALLE or POVPC was characterized by low stimulation index after immunization ox-LDL compared to control mice (PBS). Since the induction of immunomodulation by oral administration like nasal introduction had no significant effect on other measured parameters, such as weight gain, levels of triglycerides or cholesterol in plasma or immunocompetent (see tables 1, 2 and 3), the results obtained indicate the specific suppression of immune reactivity against ox-LDL. Thus, oral administration of synthetic L-ALLE, which is a component of oxidized LDL, is an effective method of reducing cellular immune responses to immunogenic and atherogenic components of plaques in genetically prednisolone the different mice APO-E KO. Figure 4 also shows a similar, but less effective inhibition of proliferation when administered orally less resistant synthetic POVPC, which is a component of LDL.

Example VI

Inhibition of atherogenesis in genetically predisposed mice (APO-E KO) as a result of oral administration of D - and L-isomers ALLE and POVPC

As it was found that oral administration of ALLE and POVPC inhibits the initial stage of atherogenesis and immune reactivity against the antigen LDL person associated with the plaques were produced comparing the ability of D - and L-isomers of essential analog LDL and non-terrestrial analogue POVPC to suppress the development of atherogenesis in older mice. Their influence on triglyceride and cholesterol in the VLDL fraction was also controlled by the method of FPLC. 57 male mice APO-E KO at the age of 24.5 weeks were divided into 5 groups. In group a (n=11) mice as described above was administered via a stomach tube 1 mg/mouse L-ALLE suspended in 0.2 ml PBS every other day for 5 days. In group b (n=9) mice as described above oral was administered 1 mg/mouse D-ALLE suspended in 0.2 ml PBS every other day for 5 days. In group C (n=10) mice as described above through the gastric tube was injected 1 mg/mouse POVPC, suspended in 0.2 ml PBS every other day for 5 days. The control group D (n=10) oral was administered PBS (containing the same amount is canola, in groups a, b, C). The basic group was slaughtered at time = 0. Subjects antigens oral was administered every 4 weeks (5 oral introductions; every other day), beginning of 24.5 weeks of age, for 12 weeks (3 series oral injections).

In mice took blood samples to oral administration (time 0), after 2 series of oral administration and at the end of the experiment (end) to determine the lipid profile, fractionation of lipids and plasma levels. Atherosclerosis was assessed as described above in the aortic sinus and the aorta. Animals were removed spleen for analysis of proliferation after 12 weeks after the first oral administration. Mass was recorded every 2 weeks throughout the experiment. All mice received normal feed, containing 4.5 wt.% fat (0.02% cholesterol), and water needs.

P=0,481
Table 5
Inhibition of atherogenesis in mice Aro-E TO the result of oral administration of L-ALLE, D-ALLE and POVPC
Period of timeMeasured valuePBS (n=10)L-ALLE 1 mg (n=11)D-ALLE 1 mg (n=9)POVPC 1 mg (n=10)Base group (t=0 (n=8) The statistical-
the cue data
Time 0Weight28,1±0,529±0,629,8±0,729,6±0,729,8±1,1P=0,445
Cholesterol413±27413±23409±28401±21393±16P=0,976
Triglyceride67±563±863±467±771±8P=0,946
EndWeight28,5±0,629,7±0,530,4±0,829,9±0,5-P=0,177
Cholesterol365±15391±18394±15358±28-
Triglyceride84±483±494±485±3-P=0,207
The lesions in the aortic sinus (μm2)369688± 32570233056± 12746245938± 20474245750± 20423225714± 5869P<0,001
The aorta
(% of
the total area)
4,5of 5.44,58,31,4P=0.002
Note: the “mass” means the mass in grams; cholesterol” means the level of cholesterol in plasma and “triglyceride” refers to the level of triglycerides in plasma, expressed in mg/DL.

As shown in figure 7, the results in table 5 indicate that effective inhibition of a late stage of atherogenesis, measured in tissues of older mice after prolonged oral administration of D - and L-isomers ALL and POVPC dose, equal to 1 mg/mouse, compared with the control mice that oral was administered PBS. Oral administration of these compounds had no significant effect on other measured parameters, such as weight gain, levels of total triglycerides or cholesterol in the plasma. Thus, synthetic D, L-ALLE and POVPC, are components of oxidized LDL have antiatherogenic activity, almost completely protected from the development of atherogenesis (compared with estimates of the lesion at the age of 24.5 weeks) genetically predisposed mice APO-E KO. Very surprising is the fact that the inhibition of atherogenesis analogs of oxidized LDL is accompanied by a significant decrease in the levels of VLDL cholesterol and triglycerides measured using FPLC (figures 8 and 9).

Example VII

Inhibition of atherogenesis in genetically predisposed mice (APO-E KO) as a result of oral administration of CI-201

We studied the ability to suppress atherogenesis sustainable form phospholipid esterified acid derivative ALLE, CI-201. Male mice APO-E KO at the age of 12 weeks were divided into two groups. In group a (n=14) mice orally was administered via a stomach tube CI-201 (0.025 mg/dose), suspended in PBS, every day for 8 weeks (5 times a week). Mice in group b (n=15) were administered PBS (control group). Atherosclerosis was assessed by you is opisannym way. All mice received normal feed, containing 4.5 wt.% fat (0.02% cholesterol), and water needs.

As shown in figure 11, the results indicate a significant slowing the development of atherosclerosis in the tissues of mice that oral was administered low doses of CI-201, compared to unimmunized control mice (PBS). The lesions in the aortic sinus in the group treated with CI-201, was equal 125192±19824 μm2and in the control group (treated with PBS) was equal 185400±20947 μm2that points to 33% (P=0,051) reduction of lesions in the aortic sinus by oral administration of low doses of CI-201. The expression of IL-10 (determined by the method of RT-PCR) in the aorta was 40% higher in the group treated with CI-201, compared with the control group. Elevated levels of expression of IL-10 in the target organ, the aorta, confirm the anti-inflammatory effect induced by oral administration of CI-201. Thus, a stable synthetic CI-201 oxidized LDL induces immunomodulation and anti-inflammatory effect when administered orally.

Example VIII

Expression of cytokines in the aorta of mice APO-E KO immunized with oxidized phospholipids (ALLE, CI-201, Et-acetal, IU-acetal and oxLDL)

The influence of ALLE, CI-201, its corresponding acetylenic derived Et-acetal and Me-acetal (compound IIa and IIb, figure 10) and oxLDL on the expression of cytokines in the body-the mission is, the aorta was determined by the method described above RT-PCR. Mice Aro-E TO oral was administered 1 mg/mouse ALLE, 1 mg/mouse CI-201, 1 mg/mouse Et-acetal, 1 mg/mouse IU-acetal, 0.1 mg/mil oxLDL or 0.2 ml/mil PBS. Oral administration produced 5 times a day. The expression of anti-inflammatory cytokine IL-10 and Pro-inflammatory cytokines IFN-γ and IL-12 were determined 8 weeks after the last oral administration.

As shown in figures 12A and 12b, in mice which were injected ALLE, CI-201, Et-acetal, IU-acetal and oxLDL were observed to have elevated levels of expression of IL-10 compared with the control group treated with PBS. As shown in figures 12C and 12d, there was observed the opposite effect on the expression level of IFN-γ and IL-12. Decreased levels of expression of IFN-γ were detected in mice that were injected ALLE, CI-201, BF-acetal and oxLDL, and decreased levels of IL-12 were detected in mice that were injected ALLE, CI-201, Et-acetal and oxLDL.

Example IX

Inhibition of atherogenesis in mice LDL-RD in the induction of immunomodulation by oral administration oxLDL

To demonstrate that the above synthetic oxidized phospholipids induce the same effect as oxidized LDL person, a model was created to assess the impact of oxLDL on the development of atherosclerosis in mice.

Mice LDL-RD aged 8-12 weeks were divided into different groups according to age, body weight and lipid profile (olstein and triglyceride). Each group was injected oxLDL in increasing doses (10, 100 or 1000 mg/dose, dissolved in PBS in a total volume of 0.2 ml PBS), albumin (100 μg/dose, dissolved in PBS in a total volume of 0.2 ml PBS) or PBS (0.2 ml) 5 times a day. The day after the last oral administration mice were given according to the needs of food, contributing to the development of atherosclerosis (“Western diet”), and within five weeks contained in 12-hour cycle of alternation of light and darkness.

Mice were killed after 6.5 weeks after the first oral administration and determined the area of atherosclerotic plaques in the aortic sinus as described above.

The effect of oxLDL on the metabolic rates are shown in the following table 6. As shown in table 6, oxLDL, without affecting the body weight or cholesterol levels, in a dose of 100 mcg/dose significantly (P<0,05) reduced triglyceride levels compared to control groups, which were administered PBS and albumin.

Table 6
The impact of the introduction of oxLDL on the metabolic rates in mice LDL-RD
Period of timeIndexGroup a
OxLDL
1000 ug/dose
Group
OxLDL
100
mcg/dose
The group With OxLDL 10 μg/dose Group D saworo-accurate human albuminGroup
E
PBS
Statistics-
tices-
Kie
data
T=0Weight (g)28,1±0,627,3±0,626,9±0,528,5±0,627,9±0,5N.S.*
Cholesterol (mg/DL)161±10160±8156±9163±10162±7N.S.*
Triglyceride
(mg/DL)
154±12145±11152±10140±14155±12N.S.*
EndWeight29,0±0,726,8±0,427,6±0,729,4±1,829,1±0,6N.S.*
Cholesterol (mg/DL)1541±175 1372±1221458±1011589±761554±121N.S.*
Triglyceride
(mg/DL)
344120250303306P<0,001**
* N.S. not statistically significant value
** Was performed by analysis of variance of rank correlations according to one characteristic of the Kruskal Wallis, data are presented as mean values.
Note: the “mass” means the mass in grams; cholesterol” means the level of cholesterol in plasma and “triglyceride” refers to the level of triglycerides in plasma, expressed in mg/DL.

Slow atherogenesis by oral administration OxLDL shown in figure 13 and in table 7. As shown in figure 13, the injection of 100 μg/dose and 1000 μg/dose oxLDL significantly reduces (P<0.001) lesions in the aortic sinus by 45% compared to control groups (which were administered PBS or serum albumin human (HAS)).

Table 7
The impact of the introduction of OxLDL on the horse atherosclerotic lesions in the aortic sinus
IndexGroup And OxLDL 1000 μg/doseGroup In OxLDL 100 mcg/doseThe group With OxLDL
10 μg/dose
Group D serum albumin manGroup E PBSStatistics-
tices-
Kie
data
Defeat
in the sinus
aorta (μm2)
37750± 489038304± 444345568± 330977604± 503969712± 6797P<0,001

Example X

Inhibition of atherogenesis in genetically predisposed mice (APO-E KO) as a result of oral administration of CI-201

As a model prevent the development of atherogenesis used mice APO-E KO at the age of 26-28 weeks (ARO-E -/-<tm1Unc> [C57B6J]). Mice were divided into different groups according to age, body weight and lipid profile (cholesterol and triglyceride). One group, which was slaughtered in the beginning of the experiment, served as the “base” group. All other groups orally was administered increasing doses of CI-201 (0.1, 1 or 10 μg/dose, rastvornogo in PBS containing 0.05% ethanol in a total volume of 0.2 ml PBS). The control group was administered PBS (0.05% ethanol, 0.2 ml).

We the am was introduced CI-201 or PBS in the form of three series at the beginning of each month, with each series consisted of 5 oral introductions, running through the day. All mice received normal feed, containing 4.5 wt.% fat (0.02% cholesterol), and water needs, while mice were kept in 12-hour cycle of alternation of light and darkness.

After 12 weeks, mice were killed and opredelyali size of atherosclerotic plaques in the aortic sinus as described above.

The impact of the introduction of CI-201 on the metabolic rates shown in table 8. The results show that CI-201 does not affect the body weight and lipid profile in experimental mice.

Table 8
The impact of the introduction of CI-201 on the metabolic rates in mice APO-E KO
Period of timeIndexGroup a CI-201 10 μg/doseGroup CI-201 1 μg/doseThe group With CI-201 0.1 µg/doseGroup D PBSBand E basicStatistics-
RCM data
T=0Weight (g)26,6±0,526,4±0,426,6±0,426,5±0,4 26,7±0,5N.S.*
Cholesterol (mg/DL)321±23323±20313±14316±12315±20N.S.*
Triglyceride (mg/DL)88±487±487±680±484±8N.S.*
EndWeight29,6±0,529,0±0,428,8±0,728,1±1,8-N.S.*
Cholesterol (mg/DL)344±25382±24406±39354±24-N.S.*
Triglyceride (mg/DL)76±496±8107±1391±7- N.S.*
* N.S. not statistically significant value
Note: the “mass” means the mass in grams; cholesterol” means the level of cholesterol in plasma and “triglyceride” refers to the level of triglycerides in plasma, expressed in mg/DL.

Slow atherogenesis by oral administration of CI-201 shown in figures 14a-b and needlemouse table 9.

Table 9
The effect of CI-201 in the area of atherosclerotic lesions in the aortic sinus
IndexGroup a CI-201 10 μg/doseGroup CI-201
1 mcg/dose
The group With CI-201 0.1 µg/doseGroup D
PBS
Band E basicStatistical
data
The lesions in the aortic sinus (μm2)272483±
20505
295729±
20909
228000±
25772
328491±
21920
218602±
29248
P<0,005

The results in table 9 show that the introduction of CI-201 completely inhibits the development of the disease, while the horses lesions in the aortic sinus of mice, which were administered different doses of CI-201, were similar to lesions in the base group.

In contrast, in mice that were administered PBS, was observed 50% is statistically significant (p<0.01) increase in atherosclerotic lesions in the aortic sinus compared with the reference group (328491±21920 μm2in the group which was administered PBS, compared to 218602±29248 μm2in the base group).

As shown in figure 14a, all doses of CI-201 inhibited disease development, the most effective dose was the minimum dose of 0.1 μg/dose. As shown in figure 14b, the group that was injected with 0.1 μg/dose of CI-201, was observed in 92% statistically significant (P<0.05) reduction of atherosclerotic lesions in the aortic sinus compared with the group which was administered PBS (328491±21920 μm2in the group treated with PBS, compared with 228000±25772 μm2in the group treated with CI-201).

Example XI

The increase of inflammatory markers in the serum of mice APO-E KO immunized with CI-201

Given the significant inhibition of atherosclerosis by oral administration of CI-201, which, as described above, is not accompanied induced change in body weight or lipid profile, investigated the effect of oral administration of CI-201 on the levels of inflammatory markers in serum to study its mechanism of action.

As established in the experimental and liniceski research IL-10 is a major protective cytokine growth and stability of the plaque. For example, Caligiuri other (Caligiuri et al., Interleukin-10 deficiency = MKD increases atherosclerosis, thrombosis, and low-density lipoproteins in apolipoprotein E knockout mice. Mol. Med. 2003; 9(1-2):10-17) recently reported that the magnitude of the lesion significantly increased in mice with absence of the APO-E and IL-10 compared with control mice, with increased proteolytic and procoagulant activity, indicating that IL-10 can reduce atherosclerosis and improves the stability of the plaque.

Another marker of acute inflammatory condition is serum amyloid A (SAA), a highly sensitive marker of inflammation, the amount of which can increase 1000-fold during inflammation. SAA as well as CRP (C-reactive protein) is synthesized by the liver under the influence of IL-1, IL-6 and TNF (Balke and Ridker, Novel clinical markers of vascular wall inflammation, Circ. Res. 2001; 89:763-771). Found that SAA is expressed in place of atherosclerotic lesion cells of several types (Meek et al., Expression of apolipoprotein serum amyloid A mRNA in human atherosclerotic lesions and cultured vascular cells: implications for serum amyloid A function. Proc. Natl. Acad. Sci. USA 1994; 91:3186-3190; Uhlar and Whitehead. Serum amyloid A, the major vertebrate acute-phase reactant. Eur. J. Biochem 1999; 265:501-523).

Inflammation initiate primarily activated blood monocytes and tissue macrophages in the location of the stimulus that causes inflammation. Activated macrophages secrete a number PE the primary mediators of inflammation, the most important of which are members of the families of cytokines IL-1 and TNF, which trigger the release of a whole series of secondary cytokines and chemokines (IL-6, IL-8 and MCP). Chemotactic activity of these molecules directs leukocytes to the site of inflammation, where they in turn produce other Pro-inflammatory cytokines.

Mice Aro-E TO oral was administered 0.1 mg/mouse CI-201 or 0.2 ml/mouse in pBS 5 times a day. In mice took the serum before the start of the experiment (day 0), at the end of the experiment (two weeks) and two weeks after the end of the experiment (4 weeks) and determined the levels of inflammatory markers IL-10 and serum amyloid A (SAA).

The obtained data are presented in figure 15A (for levels of IL-10) and figure 15b (SAA levels).

As shown in figure 15A, the end of the experiment (2 weeks) showed a significant increase in the levels of IL-10 in serum, and after another 2 weeks (4 weeks) revealed a decrease in levels. In the control group, which was administered PBS, throughout the experiment there were no recorded changes in the levels of IL-10 in serum.

As shown in figure 15b, the levels of SAA in serum was significantly increased in the control group, while no change was observed levels of SAA in serum in the group, which was introduced CI-201.

The results obtained clearly show that the increase in the levels of IL-10 in serum of CI-201 and duceret anti-inflammatory response which can suppress Pro-inflammatory response, which showed elevated levels of SAA. Systemic inflammation, characterized by high levels of SAA, can stimulate the destabilization of atherosclerotic plaques in addition to possible direct effects on atherogenesis. These results also confirm more direct impact CI-201 on inflammatory processes.

Example XII

Expression of cytokines in different organs of mice APO-E KO immunized with CI-201

The impact of immunization CI-201 on expressio cytokine in the target organ, the aorta, as well as in the spleen, liver, kidney and small intestine were determined using the above RT-PCR method. Mice Aro-E TO oral was administered 1 mg/dose of CI-201 or 0.2 ml/mouse in PBS 5 times a day. The expression of anti-inflammatory cytokine IL-10 and Pro-inflammatory cytokine IFN-γ was determined in 8 weeks after the last oral administration. The data obtained is shown in figures 16A-b and on figure 17.

As shown in figures 16A and 16b, in mice which were injected CI-201, was observed elevated levels of anti-inflammatory cytokine IL-10 compared with the control group, which was administered PBS, and the opposite effect, namely a reduced expression level, was charakteren for proinflammatory cytokine IFN-γ in the group, which was introduced CI-201.

Strengthening protivovospalitel the Noah reaction, as evidenced by elevated levels of IL-10, accompanied by a weakening of the Pro-inflammatory response, as evidenced by reduced levels of IFN-γ, emphasizes the role of immunomodulation induced CI-201, which is expressed in the switching response from Th1 to Th2 in the aorta, as well as its anti-inflammatory effect.

While CI-201 amplifies the inflammatory response in the target organ, the aorta, this effect is not present in other organs. As shown in figure 17, there is no difference in the expression of cytokines in the spleen and small intestine between the group, which was introduced CI-201, and the control group, which was administered PBS. This suggests that orally administered antigen affects Peyer's plaques are in the specified authority. In liver and kidney was not observed changes in the expression of cytokines (data not shown).

The results obtained above suggest that the analogs of oxidized phospholipids according to the present invention inhibit atherosclerosis by activating a way that affects both the immune system and the inflammation site. However, it is possible that other mechanisms are also involved in the achievement of the strongest inhibitory effect.

Example XIII

Inhibition of rheumatoid arthritis in rats suffering from adjuvant-induced arthritis, the result of the E. oral administration of CI-201

Rheumatoid arthritis (RA) is a severe autoimmune disease characterized by chronic inflammation and destruction of joints. Adjuvant-induced arthritis (AIA) is the first experimental model of arthritis (Pearson. Development of arthritis, periarthritis and periostitis in rats given adjuvant. Proc. Soc. Exp. Biol. Med. 1956; 91:95-101; Pearson and Wood. Studies of polyarthritis and other lesions induced in rats by injection of mycobacterial adjuvant. I. general clinical and pathologic characteristics and some modifying factors. Arthritis Rheum 1959; 2: 440-459).

The morphology of the lesions at the initial stage of the AIA defines cell-mediated immunity (CMI). For infiltration of lymphocytes should swelling, deposition of fibrin and necrosis, accompanied by proliferation of synoviocytes and fibroblasts and activation of osteoblasts and osteoclasts. Inflammatory infiltrate in areas of joint damage AIA contains T-cells activated by specific antigens. The Th1 cytokines, such as IL-17, IFN-γ and TNF-α, expressed in the initial stage, AIA, together with cytokines characteristic of activated macrophages. At a later stage of the disease increased levels of IL-4, IL-6 and JE (homolog protein 1 chemoatractant monocytes mouse) and TGF-β. Local secretion of proteolytic enzymes and/or oxygen radicals, which cause gradual destruction of collagen types II and IX, matrix and eventually bone (Van Eden and Waksman. Immune regulation in adjuvant-induced arthritis. Possible implicatons for innovative therapeutic strategies in arthritis. Arthritis Rheum. 2003; 48(7):1788-1796).

Several attempts immunotherapy of autoimmune diseases such as rheumatoid arthritis (RA), diabetes type I and multiple sclerosis, by modulation of the individual immune pathways involved in inflammation, or acquisition of tolerance to various antigens has shown that this approach may be promising (Bielekova et al. Encephalitogenic potential of the myelin basic protein peptide (amino acids 83-99) in multiple sclerosis: Results of a phase II clinical trial with an altered peptide ligand.Nat Med.2000;6:1167-1175; Kappos et al. Induction of a non-encephalitogenic type 2 T helper-cell autoimmune response in multiple sclerosis after administration of an altered peptide ligand in a placebo-controlled, randomized phase II trial.Nat. Med.2000;6:1176-1182).

Many of the subjects of rheumatoid arthritis is accompanied by cardiovascular diseases, and these diseases are the main cause of increased mortality. It is believed that autoantibodies against cardiolipin (CL) and modified by the oxidation of low-density lipoprotein (LDL, oxidized copper), including LDL modified by malondialdehyde (MDA-LDL), can serve to predict cardiovascular disease. It was determined that subjects suffering from rheumatoid arthritis, there are elevated levels of autoantibodies against oxidized copper low-density lipoprotein, malondialdehyde-modified low-density lipoprotein and cardiolipin (Cvetkovic et al. Increased levelsof autoantibodies are against copper-oxidized low density lipoprotein, malondialdehyde-modified low density lipoprotein and cardiolipin in patients with rheumatoid arthritis. Rheumatology. 2002; 41:988-995). In addition, there are data on the presence of oxidized low density lipoprotein in the synovial fluid of subjects suffering from rheumatoid arthritis (Dai et al. Evidence for oxidized low density lipoprotein in synovial fluid from rheumatoid arthritis patients. Free It. Res. 2000; 32(6): 479-486).

As it was found that CI-201 can effectively modulate the immune response to Ox LDL and increase anti-inflammatory reaction were investigated its influence on the development of arthritis.

The male mice Lewis at the age of nine weeks oral was administered different doses of CI-201 (4 mg/kg or 0.4 mg/kg) or PBS every other day for 5 times. Then caused adjuvant-induced arthritis using intradermal injections of 0.1 ml suspension of tuberculosis microbacteria. The severity of arthritis was monitored by measuring the swelling of the paws and mobility of animals. Study design is shown in figure 18. The results are shown in figure 19.

As shown in figure 19, the preliminary introduction of a higher dose (4.0 mg/kg) CI-201 caused a significant decrease in the swelling of the paws of rats compared with control rats, which were administered PBS.

While rats that were administered PBS, hardly moved using only the hind legs, the mobility of rats which had previously introduced a higher dose of CI-201, was close to the mobility Zdor the o rats.

To assess the effect of continuous administration of CI-201 health AIA-induced rats Lewis male Lewis rats aged 9 weeks before the induction of AIA with an intradermal injection of 0.1 ml suspension of tuberculosis microbacteria 5 times a day orally was administered to the specified drug and continued to constantly enter the specified medication three times a week for 30 days. Study design is shown in figure 20. The results are shown in figures 21-23.

As shown in figures 21-23, the continuous introduction of high doses of CI-201 significantly slowed down the development of arthritis in all investigated parameters.

The obtained results clearly show that addition of atherosclerosis CI-201 can have an anti-inflammatory effect on the classical inflammatory disease of RA.

CD4+ T cells-helper cells and macrophages penetrate through the synovial membrane (SM) in the case of chronic, destructive rheumatoid arthritis and probably play a major role in the promotion and preservation of the disease process. Of CD4+ T cells can be distinguished Th1 subpopulation characterized by the predominant production of IFN-γ. The predominance of Th1-cells proinflammatory type is a condition of occurrence of RA (Schmidt-Weber et al. Cytokine gene activation in synovial membrane, regional lymth nodes, and spleen during the course of rat adjuvant arthritis. Cell. Immunol. 1999; 195:53-65). Macrophages also strongly activated in Pospolita the EBM process, due to rheumatoid arthritis (RA), both locally and systemically.

The similarity of the inflammatory response that occurs in atherosclerosis and arthritis, confirms the assumption that CI-201 induces an inflammatory reaction in the case of AIA, a similar reaction as that described above for atherosclerosis.

Therefore, we can conclude that the introduction of CI-201 causes an increase in IL-10 level in the AIA model, in turn, IL-10 can suppress Pro-inflammatory cytokines, thus weakening the course of the disease, as demonstrated less swelling of the legs and the best mobility. Therefore, the results suggest that the analogs of oxidized phospholipids according to the present invention can serve as a new family of therapeutic agents for the treatment of rheumatoid arthritis and other autoimmune and/or inflammatory diseases.

Example XIV

Oral administration of pre-oxidized compounds V are genetically predisposed mice (APO-E KO) - the influence of oxidized groups on the inhibition of atherogenesis

Influence of oxidized groups in and ALLE, CI-201 investigated by comparing the effects of oral administration and ALLE, CI-201 at the initial stage of atherogenesis and subsequent formation of atherosclerotic plaques with the influence of pre-oxidized derivative pointed to by the x substances, namely, compound V (1-hexadecyl-2-(5'-hexenyl)-sn-glycero-3-phosphocholine, example I).

25 female mice APO-E KO aged 8-10 weeks were divided into 4 groups. Each group orally was administered 5 mg/mouse connection V, suspended in 0.2 ml PBS (group a, n=6), 1 mg/mouse connection V”, suspended in 0.2 ml PBS (group b, n=6), 0.2 mg/mouse connection V, suspended in 0.2 ml PBS (group C, n=6), and PBS (group D, control group, n=7) every other day for 5 days. Eight weeks after the last oral administration, the mice were killed. In mice took blood samples before the start of the experiment (time 0) and at the end of the experiment (end) to determine the lipid profile. Atherosclerosis was assessed in the heart as described above. All mice received normal feed, containing 4.5 wt.% fat (0.02% cholesterol), and water needs.

The impact of the introduction of a connection V on rates of metabolism and atherogenesis shown in table 10. The influence of the connection V on atherogenesis further shown in figure 24.

Table 10
The influence of pre-oxidized derivative of compound V on the metabolic rates and the area of atherosclerotic lesions in the aortic sinus of mice APOE KO
Period of timeMeasure the th indicator PBS (n=7)Connection V 5 mg (n=6)Connection V 1 mg (n=6)Connection V 0.2 mg (n=6)Statistical data (value
R)
Time 0Weight18,4±0,418,0±0,318,3±0,318,5±0,30,826
Cholesterol197±19210±18198±23214±210,902
Triglyceride53±1052±452±550±70,993
EndWeight20,7±0,419,8±0,319,8±0,220,1±0,30,161
Cholesterol387±20431±23398±23 409±80,455
Triglyceride70±380±891±4**102±5*0,001
The lesions in the aortic sinus (μm2)314764± 14458307291± 22689361166± 24068334622± 261000,352
Note: the “mass” means the mass in grams; cholesterol” means the level of cholesterol in plasma and “triglyceride” refers to the level of triglycerides in plasma, expressed in mg/DL.
* P<0,001 compared with the group treated with PBS.
**P<0.05 compared with the group treated with PBS.

As shown in figure 24, while oral administration of oxidized compounds CI-201 and ALLE significantly inhibited atherogenesis in mice APO-E KO, the introduction of pre-oxidized derivative compounds V had no effect on atherogenesis, which shows the importance of the presence of oxidized groups in the treatment of atherogenesis.

Example XV

Oral administration of the pre is satisfactory oxidized compounds V are genetically predisposed mice (APO-E KO) - influence of oxidized groups on the development of atherogenesis

Next explored the possibility of using pre-oxidized compounds V to prevent the development of atherogenesis in the above model using mice APOE KO. Mice APO-E KO at the age of 23-26 weeks (ARO-E -/-<tm1Unc>[C57B/6J]) were divided into different groups according to age, body weight and lipid profile (cholesterol and triglyceride). One group, which was slaughtered in the beginning of the experiment, served as the “base” group (B.L., n=10). The second group was injected compound V (0,1 μg/dose, n=10), dissolved in PBS containing 0.05% ethanol in a total volume of 0.2 ml PBS. The control group was administered PBS (0.05% ethanol, 0.2 ml) (n=11).

Mice were administered compound V or PBS in the form of three series at the beginning of each month, with each series consisted of 5 oral introductions, running through the day. All mice received normal feed, containing 4.5 wt.% fat (0.02% cholesterol), and water needs, while mice were kept in 12-hour cycle of alternation of light and darkness.

After 12 weeks, mice were killed and determined the lipid profile and size of atherosclerotic plaques in the aortic sinus as described above.

The results given in needlemouse table 11 and figure 25 clearly shows that oral administration of compound V did not affect the development of atherogenesis.

Table 11
The influence of pre-oxidized derivative of compound V on the metabolic rates and the area of atherosclerotic lesions in the aortic sinus of mice APOE KO
Period of timeIndexConnection V
0.1 mg/dose
PBSBase groupStatistics (p value)
Time 0Weight (g)26,1±0,726,2±0,626,2±0,70,710
Cholesterol (mg/DL)401±27393±32406±190,936
Triglyceride (mg/DL)128±8129±6125±70,926
EndWeight (g)28,1±0,528,0±0,5-0,967
Cholesterol (mg/DL)292±16277±37-0,717
Triglyceride (mg/DL)81±390±4-0,094
The lesions in the aortic sinus (μm2)2338156± 32206206647±15293137451±189750,011*
* No statistically significant difference between mice treated with compound V, and the control mice treated with PBS.

Although the invention is described on the basis of specific variants of its obshestvinie, it is clear that professionals in this field should be visible many alternatives, modifications and variations of the present invention. Therefore, the present invention applies to all such alternatives, modifications and variations that are included in the scope of the attached claims.

All publications, patents and patent applications listed in the description of the invention, and fully incorporated in the description of the invention as SS is the CTL, as if each individual publication, patent or patent application were specifically included in this description by reference. In addition, citation and identification of any reference in this application shall not be construed as an admission that such reference characterizes the prior art relative to the present invention.

Additional interest links

(not mentioned in the description of the invention)

Wick G, Schett G, Amberger A, Kleindienst R, Xu Q. atherosclerosis Is an immunologically mediated disease? Immunol Today 1995; 16: 27-33.

Libby P, Hansson GK. Involvement of the immune system in human atherogenesis: current knowledge and gets questions. Lab Invest 1991; 64: 5-15.

Steinberg D, Parathasarathy S, Carew TE, Khoo JC, Witztum JL. Beyond cholesterol. Modifications of low-density lipoprotein that increase its atherogenicity. N Engl J Med 1989; 320: 915-924.

Witztum J. The oxidation hypothesis of atherosclerosis. Lancet 1994; 344: 793-795.

Palinski W, Miller E, Witztum JL. Immunization of low density lipoprotein (LDL) receptor-deficient rabbits with homologous malondialdehyde-modified LDL reduces atherogenesis. Proc Natl Acad Sci USA. 1995;92: 821-825.

George J, Afek A, Gilburd B, Levy Y, Levkovitz H, Shaish A, Goldberg I, Kopolovic Y, Wick G, Shoenfeld Y, Harats D. Hyperimmunization of ApoE deficient mice with homologous oxLDL suppresses early atherogenesis. Atherosclerosis. 1998; 138: 147-152.

Weiner H, Freidman A, Miller A. Oral tolerance: immunologic mechanisms and treatment of animal and human organ specific autoimmune diseases by oral administration of autoantigens. Annu Rev Immunol 1994; 12: 809-837.

Palinski W, Ord VA, Plump AS, Breslow JL, Steinberg D, Witztum JL. Apo-E-deficient mice are a model of lipoprotein oxidation in atherogenesis. Demonstration of oxidation-specific epitopes in lesions and high titers of autoantibodies are to malondialdehyde-lysine in serum. rterioscler Thromb 1994; 14: 605-616

Roselaar SE, Kakkanathu PX, Daugherty A. Lymphocyte populations in atherosclerotic lesions of apo-E -/- and LDL receptor -/- mice; Decreasing density with disease progression. Arterioscler Thromb Vase Biol 1996; 16: 1013-1018.

Palinski W, Ylä-Herttuala S, Rosenfeld ME, Butler SW, Socher SA, Parthasarathy S, Curtiss LK, Witztum JL. Antisera and monoclonal antibodies specific for epitopes generated during oxidative modification of low density lipoprotein. Arteriosclerosis 1990; 10: 325-335.

Ou z, Ogamo A., L. Guo, Y. Konda, Harigaya Y. and Nakagawa Y. Anal. Biochem. 227: 289-294, 1995.

E. Baer and Buchnea JBC. 230,447,1958.

Quintana FJ, Carmi P, Mor F and Cohen R. Inhibition of adjuvant arthritis by a DNA vaccine encoding human heat shock protein 60. J. Immunol.2002; 169:3422-3428.

Cobelens PM, Heijnen CJ, Nieuwenhois EES et al. Treatment of adjuvant induced arthritis by oral administration of mycobacterial Hsp65 during disease. Arthritis Rheum. 2000;43(12):2694-2702.

Cobelens PM, Kavelaars A, Van Der Zee R, et al. Dynamics of mycobacterial HSP65-induced T-cell cytokine expression during oral tolerance induction in adjuvant arthritis. Rheumatology. 2002;41:775-779.

1. Pharmaceutical composition having immunomodulatory activity, containing as active ingredient a therapeutically effective amount of the compounds of General formula I:
,
where n is the integer 3;
In1is oxygen or sulfur;
each of In2and In3is oxygen;
each And1and a2is CH2;
Y is chosen from the group consisting of hydrogen, phosphoric acid, phosphorylcholine and phosphorylethanolamine; and
each of X1and X2independently means a saturated or unsaturated hydrocarbon of the General formula II:

where m is an integer from 2 to 19; and
Z is chosen from the group consisting of:

where W is oxygen; and
at least one of X1and X2includes Z, which is not
hydrogen; and
where each of R1, R2, R3,andis hydrogen;
each of R" and R"' are independently selected from the group consisting of hydrogen and alkyl;
each of Ra, R'a, Rb, R b,..., Rm-1, R m-1, Rm and R m is independently selected from the group consisting of hydrogen, communication and hydroxy-group; and
or its stereoisomer, optical isomer, racemic mixtures, pharmaceutically acceptable salt,
and pharmaceutically acceptable carrier,
moreover, the mentioned pharmaceutical composition is packaged in a packaging material and identified in print, made in the specified packaging material or packaging material, as intended for use in the treatment or prevention of inflammation associated with endogenous oxidized lipid, and specified the inflammation associated with the disease or disorder selected from the group consisting of inflammatory gastrointestinal diseases or disorders, proliferative diseases or disorders and diabetes.

2. The pharmaceutical composition according to claim 1, where the specified inflammatory gastro-kishen is s disease or disorder selected from the group consisting of colitis, ileitis, Crohn's disease, chronic inflammatory disease of the small bowel, syndrome of irritated large intestine, chronic inflammatory diseases of the bowel, celiac disease, ulcerative colitis, ulcers, ulcerations of the skin, sores, ulcers, peptic ulcers, ulcers cheeks, nasopharyngeal ulcers, esophageal ulcers, duodenal ulcers and gastrointestinal ulcers.

3. The pharmaceutical composition according to claim 2, where the specified inflammatory gastrointestinal disease or disorder selected from the group consisting of colitis and chronic inflammatory diseases of the large intestine.

4. The pharmaceutical composition according to claim 2, where the specified inflammatory gastrointestinal disease or disorder selected from the group consisting of Crohn's disease and ulcerative colitis.

5. The pharmaceutical composition according to claim 1, where the specified proliferative disease or disorder is a malignant tumor.

6. The pharmaceutical composition according to claim 1, where the specified disease or disorder is a diabetes.

7. Pharmaceutical composition having immunomodulatory activity, containing as active ingredient a therapeutically effective amount of the compounds of General formula:

where n is the integer 3;
In 1is oxygen or sulfur;
each of In2and In3is oxygen;
each of the A1and a2is CH2;
Y is chosen from the group consisting of hydrogen, phosphoric acid, phosphorylcholine and phosphorylethanolamine; and
each of X1and X2independently means a saturated or unsaturated hydrocarbon of the General formula II:

where m is an integer from 2 to 19; and
Z is chosen from the group consisting of:

where W is oxygen; and
at least one of X1and X2includes Z, which is not
hydrogen; and
where each of R1, R2, R3,andis hydrogen;
each of R" and R"' are independently selected from the group consisting of hydrogen and alkyl;
each of Ra, R'a, Rb, R b,..., Rm-1, R m-1, Rm and R m is independently selected from the group consisting of hydrogen, communication and hydroxy-group;
or its stereoisomer, optical isomer, racemic mixtures, pharmaceutically acceptable salt,
and pharmaceutically acceptable carrier,
moreover, the mentioned pharmaceutical composition is packaged in a packaging material and identified in print, made in the specified packaging material or packaging material, as intended to reduce titoki is a, selected from the group consisting of interleukin-6, interleukin-17, macrophage chemotactic protein-1 (MCP-1), macrophage chemotactic protein-3 (MCP-3) and tumor necrosis factor-α (TNF-α).

8. Pharmaceutical composition having immunomodulatory activity, containing as active ingredient a therapeutically effective amount of the compounds of General formula:

where n is the integer 3;
B1is oxygen or sulfur;
each of In2and In3is oxygen;
each of the A1and a2is CH2;
Y is chosen from the group consisting of hydrogen, phosphoric acid, phosphorylcholine and phosphorylethanolamine; and
each of X1and X2independently means a saturated or unsaturated hydrocarbon of the General formula II:

where m is an integer from 2 to 19; and
Z is chosen from the group consisting of:

where W is oxygen; and
at least one of X1and X2includes Z, which is not
hydrogen; and
where each of R1, R2, R3,andis hydrogen;
each of R" and R"' are independently selected from the group consisting of hydrogen and alkyl;
each of Ra, R'a, Rb, R b,..., Rm-1, R m-1, Rm and R m is not avisio selected from the group consisting of hydrogen, communication and hydroxy-group; and
or its stereoisomer, optical isomer, a racemic mixture,
pharmaceutically acceptable salt,
and pharmaceutically acceptable carrier,
moreover, the mentioned pharmaceutical composition is packaged in a packaging material and identified in print, made in the specified packaging material or packaging material, as intended for use in the treatment of diseases or disorders, which is a favorable decrease in the level of cytokine selected from the group consisting of interleukin-6, interleukin-17, macrophage chemotactic protein-1 (MCP-1), macrophage chemotactic protein-3 (MCP-3) and tumor necrosis factor-α (TNF-α).

9. The pharmaceutical composition according to any one of claims 1 to 8, containing the compound, where X2includes Z are not hydrogen.

10. The pharmaceutical composition according to claim 9, where the specified Z is selected from
the group consisting of

11. The pharmaceutical composition of claim 10, where Y is phosphorylcholine.

12. The pharmaceutical composition according to claim 11, where In1represents oxygen.

13. The pharmaceutical composition according to item 12, where X2is a saturated hydrocarbon with the General formula II:

where m is the integer 3;
Z is where W is oxygen; and
each of R, Ra, R'a, Rb, R b,..., Rm-1, R m-1, Rm and R m is hydrogen.

14. The pharmaceutical composition according to item 13, where X1is a saturated hydrocarbon with the General formula II:

where m is an integer from 11 to 15;
Z is N; and
each of R, Ra, R'a, Rb, R b,..., Rm-1, R m-1, Rm and R m is hydrogen.

15. The pharmaceutical composition according to 14, where the specified connection selected from the group consisting of 1-hexadecyl-2-(4'-carboxy)butyl-sn-glycero-3-phosphocholine, 1-dodecyl-2-(4'-carboxy)butyl-sn-glycero-3-phosphocholine and their pharmaceutically acceptable salts.

16. The pharmaceutical composition according to 14, containing the compound of formula III:

or its pharmaceutically acceptable salt.

17. The pharmaceutical composition according to any one of claims 1 to 8, containing a compound selected from the group consisting of:
1-hexadecyl-2-(5'-oxapentane)-sn-glycero-3-phosphocholine;
3-hexadecyl-2-(5'-oxapentane)-sn-glycero-1-phosphocholine;
1-hexadecyl-2-(4'-carboxy)butyl-sn-glycero-3-phosphocholine;
1-hexadecyl-2-(4'-carboxy)butyl-sn-glycero-3-phosphoethanolamine;
1-hexadecyl-2-(6'-carboxy)hexanol-sn-glycero-3-phosphocholine;
1-hexadecyl-2-(6'-carboxy)hexanol-sn-glycero-3-phosphoethanolamine;
1-hexadecyl-2-(3'-carboxy)propyl-sn-glycero-3-phosphocholine;
1-hexadecyl-2-(3'-carboxy)propyl-sn-CH is zero-3-phosphoethanolamine;
1-hexadecyl-2-(4'-carboxymethyl)butyl-sn-glycero-3-phosphocholine;
1-hexadecyl-2-(5',5'-dioxyphenyl)-sn-glycero-3-phosphocholine;
1-hexadecyl-2-(5',5'-dimethoxyphenyl)-sn-glycero-3-phosphocholine;
1-hexadecyl-2-(5',6'-dihydroxy)hexyl-sn-glycero-3-phosphocholine;
1-dodecyl-2-(4'-carboxy)butyl-sn-glycero-3-phosphocholine;
1-hexadecyl-2-(4'-carboxy)butyl-sn-glycero-3-phosphate;
1-hexadecyl-2-(4'-carboxy)butylpyrazine;
1-(CIS-9-hexadecynyl)-2-(4'-carboxy)butyl-sn-glycero-3-phosphocholine;
1-(15'-carboxy)pentadecyl-2-(4'-carboxy)butyl-sn-glycero-3-phosphocholine;
1 -(15'-carboxy)pentadecyl-2-(4'-carboxy)butyl-sn-glycero-3-phosphoethanolamine;
1-S-hexadecyl-2-(4'-carboxy)butyl-sn-glycero-3-phosphocholine;
1-S-hexadecyl-2-(4'-carboxy)butyl-sn-glycero-3-phosphoethanolamine;
and their pharmaceutically acceptable salts.

18. A method of treating or preventing inflammation associated with endogenous oxidized lipid, and specified the inflammation associated with the disease or disorder selected from the group consisting of inflammatory gastrointestinal diseases or disorders, proliferative diseases or disorders and diabetes, and the method comprises the administration to a subject in need, a therapeutically effective amount of at least one oxidized lipid of General formula I:

where n is C the small number 3;
In1is oxygen or sulfur;
each of In2and In3is oxygen;
each And1and a2independently selected from the group consisting of CH2and C=O;
Y is chosen from the group consisting of hydrogen, phosphoric acid, phosphorylcholine and phosphorylethanolamine; and
each of X1and X2independently means a saturated or unsaturated hydrocarbon of the General formula II:

where m is an integer from 1 to 26; and
Z is chosen from the group consisting of:

where W is oxygen; and
at least one of X1and X2includes Z are not hydrogen; and
where each of R1, R2, R3,andis hydrogen;
each of R" and R"' are independently selected from the group consisting of hydrogen and alkyl;
each of Ra, R'a, Rb, R b,..., Rm-1, R m-1, Rm and R m is independently selected from the group consisting of hydrogen, communication and hydroxy-group;
or its stereoisomer, optical isomer, racemic mixtures, pharmaceutically acceptable salt,
thus, carrying out the treatment or prevention of the specified entity of inflammatory diseases or disorders associated with an endogenous oxidized lipid.

19. The method according to p, where the specified inflammatory gastro-Ki is acnee a disease or disorder selected from the group consisting of colitis, ileitis, Crohn's disease, chronic inflammatory disease of the small bowel, syndrome of irritated large intestine, chronic inflammatory diseases of the bowel, celiac disease, ulcerative colitis, ulcers, ulcerations of the skin, sores, ulcers, peptic ulcers, ulcers cheeks, nasopharyngeal ulcers, esophageal ulcers, duodenal ulcers and gastrointestinal ulcers.

20. The method according to claim 19, where the specified inflammatory gastrointestinal disease or disorder selected from the group consisting of colitis and chronic inflammatory diseases of the large intestine.

21. The method according to claim 19, where the specified inflammatory gastrointestinal disease or disorder selected from the group consisting of ulcerative colitis and Crohn's disease.

22. The method according to p, where the specified proliferative disease or disorder is a malignant tumor.

23. The method according to p, where the specified disease or disorder is a diabetes.

24. The way to reduce the level of cytokine selected from the group consisting of interleukin-6, interleukin-17, macrophage chemotactic protein-1 (MCP-1), macrophage chemotactic protein-3 (MCP-3) and tumor necrosis factor-α (TNF-α) in the subject, and the method includes the introduction to the subject an effective amount of at least one okislennaya General formula I:

where n is the integer 3;
In1is oxygen or sulfur;
each of In2and In3is oxygen;
each And1and a2independently selected from the group consisting of CH2and C=O;
Y is chosen from the group consisting of hydrogen, phosphoric acid, phosphorylcholine and phosphorylethanolamine; and
each of X1and X2independently means a saturated or unsaturated hydrocarbon of the General formula II:

where m is an integer from 1 to 26; and
Z is chosen from the group consisting of:

where W is oxygen; and
at least one of X1and X2includes Z are not hydrogen; and
where each of R1, R2, R3,andis hydrogen;
each of R" and R"' are independently selected from the group consisting of hydrogen and alkyl;
each of Ra, R'a, Rb, R b,..., Rm-1, R m-1, Rm and R m is independently selected from the group consisting of hydrogen, communication and hydroxy-group;
or its stereoisomer, optical isomer, racemic mixtures, pharmaceutically acceptable salts.

25. A method of treating diseases or disorders in which is a favorable decrease in the level of cytokine selected from the group consisting of interleukin-6, interleukin-17, mo is alternova chemotactic protein-1 (MCP-1), macrophage chemotactic protein-3 (MCP-3) and tumor necrosis factor-α (TNF-α), which includes an introduction to the subject an effective amount of at least one oxidized lipid of General formula I:

where n is the integer 3;
In1is oxygen or sulfur;
each of In2and In3is oxygen;
each And1and a2independently selected from the group consisting of CH2and C=O;
Y is chosen from the group consisting of hydrogen, phosphoric acid, phosphorylcholine and phosphorylethanolamine; and
each of X1and X2independently means a saturated or unsaturated hydrocarbon of the General formula II:

where m is an integer from 1 to 26; and
Z is chosen from the group consisting of:

where W is oxygen; and
at least one of X1and X2includes Z are not hydrogen; and
where each of R1, R2, R3,andis hydrogen;
each of R" and R"' are independently selected from the group consisting of hydrogen and alkyl;
each of Ra, R'a, Rb, R b,..., Rm-1, R m-1, Rm and R m is independently selected from the group consisting of hydrogen, communication and hydroxy-group;
or its stereoisomer, optical isomer, racemic mixtures, pharmaceutically when mlemos salt.

26. The method according to any of PP-25, where at least one of A1and a2is CH2.

27. The method according to p, where a2is CH2.

28. The method according to p, where a1and a2is CH2.

29. The method according to any of PP-25, where X2includes Z are not hydrogen.

30. The method according to clause 29, where the specified Z is selected from the group consisting of

31. The method according to item 30, where Y is phosphorylcholine.

32. The method according to p, where1is oxygen.

33. The method according to p, where X2is a saturated hydrocarbon with the General formula II:

where m is the integer 3;
Z is,
where W is oxygen; and
each of R, Ra, R'a, Rb, R b,..., Rm-1, R m-1, Rm and R m is hydrogen.

34. The method according to p, where X1is a saturated hydrocarbon with the General formula II:

where m is an integer from 11 to 15;
Z is N; and
each of R, Ra, R'a, Rb, R b,..., Rm-1, R m-1, Rm and R m is hydrogen.

35. The method according to clause 34, where specified, the oxidized lipid is selected from the group consisting of 1-hexadecyl-2-(4'-carboxy)butyl-sn-glycero-3-phosphocholine, 1-dodecyl-2-(4'-carboxy)butyl-sn-glycero-3-phosphocholine and their pharmaceutically acceptable salts.

36. The method according to clause 34, where the specified oxidized lipid represents Obedinenie formula III:

or its pharmaceutically acceptable salt.

37. The method according to any of PP-25, where specified, the oxidized lipid is selected from the group consisting of: 1-Palmitoyl-2-azelaoyl-sn-glycero-3-phosphocholine, 1-hexadecyl-2-azelaoyl-sn-glycero-3-phosphocholine, 1-Palmitoyl-2-glutaryl-sn-glycero-3-phosphocholine (PGPC), 1-Palmitoyl-2-(5-oxovalerate)-sn-glycero-3-phosphocholine (POVPC), 1-Palmitoyl-2-(9-Oconnor)-sn-glycero-3-phosphocholine, 1-hexadecyl-2-acetyl-sn-glycero-3-phosphocholine, 1-octadecyl-2-acetyl-sn-glycero-3-phosphocholine, 1-hexadecyl-2-buterol-sn-glycero-3-phosphocholine, 1-octadecyl-2-buterol-sn-glycero-3-phosphocholine, 1-Palmitoyl-2-acetyl-sn-glycero-3-phosphocholine, 1-octadecenyl-2-acetyl-sn-glycero-3-phosphocholine, 1-hexadecyl-2-(homogametic)-sn-glycero-3-phosphocholine, 1-hexadecyl-2-arachidonoyl-sn-glycero-3-phosphocholine, 1-hexadecyl-2-eicosapentaenoic-sn-glycero-3-phosphocholine, 1-hexadecyl-2-docosahexaenoyl-sn-glycero-3-phosphocholine, 1-octadecyl-2-methyl-sn-glycero-3-phosphocholine, 1-hexadecyl-2-butanoyl-sn-glycero-3-phosphocholine, Lyso C16 PAF, Lyso PAF C18, 1-On-1'-(Z)-hexadecanoyl-2-[12-[(7-nitro-2-1,3-benzoxadiazole-4-yl)amino]dodecanoyl]-sn-glycero-3-phosphocholine, 1-On-1'-(Z)-hexadecanoyl-2-oleoyl-sn-glycero-3-phosphocholine, 1-On-1'-(Z)-hexadecanyl-2-arachidonoyl-sn-glycero-3-phosphocholine, 1-O-1'-(Z)-hexadecanoyl-2-docosahexaenoyl-sn-glycero-3-phosphocholine, 1-O-1'-(Z)-hexa is azenil-2-oleoyl-sn-glycero-3-phosphoethanolamine, 1-O-1'-(Z)-hexadecanoyl-2-arachidonoyl-sn-glycero-3-phosphoethanolamine, 1-On-1'-(Z)-hexadecanoyl-2-docosahexaenoyl-sn-glycero-3-phosphoethanolamine, 1-S-hexadecyl-2-(4'-carboxy)butyl-sn-glycero-3-phosphocholine and 1-S-hexadecyl-2-(4'-carboxy)butyl-sn-glycero-3-phosphoethanolamine.

38. The method according to p, which additionally includes the introduction of a specified subject a therapeutically effective amount of at least one additional compound capable of treating or preventing a specified inflammation, where said additional compound is an inhibitor of HMGCoA reductase inhibitor (statin).

39. The method according to any of PP-25, where specified, the oxidized lipid is selected from the group consisting of:
1-hexadecyl-2-(5'-oxapentane)-sn-glycero-3-phosphocholine;
3-hexadecyl-2-(5'-oxapentane)-sn-glycero-1-phosphocholine;
1-hexadecyl-2-(4'-carboxy)butyl-sn-glycero-3-phosphocholine;
1-hexadecyl-2-(4'-carboxy)butyl-sn-glycero-3-phosphoethanolamine;
1-hexadecyl-2-(6'-carboxy)hexanol-sn-glycero-3-phosphocholine;
1-hexadecyl-2-(6'-carboxy)hexanol-sn-glycero-3-phosphoethanolamine;
1-hexadecyl-2-(3'-carboxy)propyl-sn-glycero-3-phosphocholine;
1-hexadecyl-2-(3'-carboxy)propyl-sn-glycero-3-phosphoethanolamine;
1-hexadecyl-2-(4'-carboxymethyl)butyl-sn-glycero-3-phosphocholine;
1-hexadecyl-2-(5',5'-dioxyphenyl)-sn-glycero-3-phosphocholine;
1-hexadecyl-2-(5',5 dimethoxyphenyl)-sn-glycero-3-phosphocholine;
1-hexadecyl-2-(5',6'-dihydroxy)hexyl-sn-glycero-3-phosphocholine;
1-dodecyl-2-(4'-carboxy)butyl-sn-glycero-3-phosphocholine;
1-hexadecyl-2-(4'-carboxy)butyl-sn-glycero-3-phosphate;
1-hexadecyl-2-(4'-carboxy)butylpyrazine;
1-(CIS-9-hexadecynyl)-2-(4'-carboxy)butyl-sn-glycero-3-phosphocholine;
1-(15'-carboxy)pentadecyl-2-(4'-carboxy)butyl-sn-glycero-3-phosphocholine;
1-(15'-carboxy)pentadecyl-2-(4'-carboxy)butyl-sn-glycero-3-phosphoethanolamine;
1-S-hexadecyl-2-(4'-carboxy)butyl-sn-glycero-3-phosphocholine; 1-S-hexadecyl-2-(4'-carboxy)butyl-sn-glycero-3-phosphoethanolamine;
and their pharmaceutically acceptable salts.



 

Same patents:

FIELD: chemistry.

SUBSTANCE: invention relates to a method of producing a lipid preparation which contains phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS) and phosphatidylinositol (PI), and sphyngomyelin (SM), a composition containing a mixture of said lipids and use thereof. The method involves steps of removing non-lipid material from bovine milk by dispersing the lipids in a mixture of a polar organic solvent and a nonpolar solvent, followed by separation of the lipid fraction and removing the solvent from said fraction, a step of de-oiling the obtained lipid fraction in order to remove nonpolar lipids by dissolving in acetone or supercritical CO2 and a step of filtering and drying the obtained lipids. In the lipid composition obtained using said method, the ratio of phospholipids is comparable with that of phospholipids in human breast milk. Disclosed is use of the composition obtained using the disclosed method as a source of human breast milk fat for making infant formula.

EFFECT: method enables to obtain a human breast milk substitute by processing readily available material and use thereof to make infant formula.

8 cl, 5 tbl, 1 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: claimed invention relates to application for production of medication for prevention and/or treatment of radiolesion or lesion of trisubstituted glycerol compound, corresponding to formula where X is selected from phosphate and sulfate; R1 is selected from C16-C20alkyl, R2 is selected from C1-C3alkyl and C1-C3hydroxyalkyl, R3 is selected from hydrogen and C1-C3 alkyl, R4 is selected from C1-C3alkyl and C3-C6cycloalkyl, R5 is selected from hydrogen and methyl, or its enantiomer, or diastereomer, or pharmaceutically acceptable salt, and at least one pharmaceutically acceptable filling agent.

EFFECT: claimed is novel application of known compounds.

12 cl, 3 dwg, 5 ex, 5 tbl

FIELD: chemistry.

SUBSTANCE: in the proposed methods, the hydroxyl group of the formula I compound is substituted with the corresponding residue so that use of column chromatography is avoided, , where A1 is CH2, R1 is alkyl, R3 is a residue of phosphoric acid, phosphorylcholine, phosphoryl ethanolamine, phosphorylserine, phosphoryl cardiolipin, phosphoryl inosite, ethylphosphocholine, phosphoryl methanol, phosphoryl ethanol, phosphoryl propanol, phosphoryl butanol, phosphoryl ethanolamine-N-lactose, phosphoethanolamine-N-[methoxy(propyleneglycol)], phosphoinosite-4-phosphate, phosphoinosite-4,5-biphosphonate, pyrophosphate, phosphoethanolaminediethylene-triaminepentaacetate, dinitrophenyl phosphoethanolamine, phosphogycerine or a reactive phosphorus-containing group which can convert to the said residues, or hydrogen.

EFFECT: design of methods of producing phospholipids, which avoid use of chromatography.

22 cl, 5 ex

FIELD: medicine.

SUBSTANCE: invention concerns the new synthetic oxidised lipids and ways of application of the oxidised lipids for treatment and inflammation prevention, associated with endogenous oxidised lipid.

EFFECT: rising of treatment efficiency.

54 cl, 25 dwg, 15 ex

FIELD: biotechnology.

SUBSTANCE: claimed method includes interaction of mixtures of natural phosphatides or components thereof, for instance, soybean or egg lecithin or animal phospholipids, or synthetic phosphatides, by reaction thereof with D phospholipase having transphosphatidylase activity in water medium.

EFFECT: simplified method for isolation of pure phosphatides of improved purity with increased yield.

28 cl, 13 ex

The invention relates to the field of medicine and for the application of hydrogenated soy phosphatidylcholine (Phospholipon-N) as an active ingredient, providing a surface-active properties of drugs for substitution of the surfactant therapy
The invention relates to an improved method for producing a water-soluble amino acid derivatives of fullerene that can be used in pharmacology and Microbiology

The invention relates to organic chemistry, namely the chemistry of fullerenes

The invention relates to new derivatives of phospholipids, namely alkylphosphates or ALCOHOLATES, kalinovy residue which is part of a heterocyclic ring, the method of obtaining a class of these compounds, as well as to medicines and the way to obtain medicines that contain compounds as biologically active substances

Antibodies // 2482131

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to immunology. There are presented the antibodies that bind to the P-selectin glycoprotein ligand-1 (PSGL-1), as well as methods for stimulating the death of activated T-cells and simulating T-cell immune response in a patient by the use of the antibodies under the invention, and pharmaceutical compositions containing the antibodies under the invention. There are also disclosed nucleic acids, expression vectors and host cells for producing the antibodies under the invention.

EFFECT: invention may can find further application in therapy of the PSGL-1 associated diseases.

35 cl, 4 ex, 3 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to biotechnology, more specifically preparing a tumour necrosis factor receptor and may be used in medicine. Genetic engineering technique is used to produce mutant TNFRp75 bound to a tumour necrosis factor and lymphotoxins substantially consisting of N-terminal 257 amino acid residues TNFRp75 wherein the N-terminal residue Glu92 is substituted by Asn, His, Ser or Ala and wherein the N-terminal residue Trp89 is optionally substituted by Tyr or Phe. The produced mutant is used to construct a fused protein with an additional amino acid fragment specified in a constant area of human immunoglobulin and one of five functional areas of albumin found on C-terminal of the soluble mutant TNFRp75. The produced mutant TNFRp75 and the fused protein is used as a part of a pharmaceutical composition for treating the diseases associated with TNFα overexpression which involve rheumatoid arthritis, psoriasis, sclerodermatitis, Sjogren syndrome, Strumpell-Marie disease, lupus erythematosus, acute disseminated myositis and syndrome similar to systemic lupus erythematosus.

EFFECT: invention enables producing soluble TNFRp75 mutant able to be bound to tumour necrosis factor and lymphotoxins at a high affinity degree.

9 cl, 12 dwg, 10 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: claimed invention relates to novel derivatives of imidazo[4,5-c]chinoline of general formula or to its pharmaceutically acceptable salts, where R1 represents straight-chained C1-C6alkyl, possibly substituted with one substituent, selected from C1-C3alkoxy; Z1 represents C2-C6alkylene; X1 represents NR5 or >NCOR5; Y1 represents C1-C6alkylene; R3 represents C1-C6alkyl, possibly substituted with C1-C6alkoxy; R5 represents hydrogen, piperidinyl, possibly substituted by piperidinyl nitrogen with group R10, group C1-C6alkyl, where the last group is possibly substituted with one substituent, independently selected from NR7R8 or R9; or R5 represents C1-C6alkylene, which can be bound with carbon atom in C2-C6alkylene group Z1 with formation of piperidine ring; each of R7 and R8 independently represents tetrahydropyranyl, piperidinyl, possibly substituted by piperidinyl nitrogen atom with group R10a, C1-C6alkyl, where the last group is possibly substituted with one group, independently selected from OR12; or R7 and R8 together with nitrogen atom, to which they are bound, form 4-7-membered saturated heterocyclic ring, selected from asetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, azepanyl, 1,4-oxazepanyl and 1,4-diazepanyl, where heterocyclic ring is possibly substituted with one or two substituents, independently selected from S(O)qR15, OR15, CO2R15, COR15, CONR15R16, NR15CO2R16, pyrimidinyl and C1-C6alkyl, where the last group is possibly substituted with one group, independently selected from OR18 and CO2R18; R9 represents S(O)qR20; R10 and R10a independently represent COR2 or group C1-C6alkyl; each of R12, R15, R16, R18, R20 and R24 independently represents hydrogen or C1-C6alkyl; q equals 2; m and n both equal 0; and A represents phenyl. Invention also relates to method of obtaining formula (I) compound, based on it pharmaceutical composition, and to method of treating said pathological conditions.

EFFECT: obtained are novel derivatives of imidazo[4,5-c]chinoline, useful modulation of TLR7 activity.

17 cl, 18 dwg, 81 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry and concerns an ergogenic formulation possessing adaptogenic, hepatoprotective and immunomodulatory action, and may be used in hepatic diseases, in decreases in immunity and increase in performance efficiency and tolerance. The composition contains burnut extract, zinc aspartate, magnesium aspartate, vitamin B6, water in certain proportions of the ingredients.

EFFECT: composition possess higher pharmacological activity and bioavailability.

3 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: group of inventions refers to medicine, namely immunology and may be used for treating rheumatoid arthritis. That is ensured by the use of a pharmaceutical composition containing an active principle presented by a monoclonal antibody which recognises a CD6 human leukocyte differentiation antigen and produced by secreting hybridoma IOR-TIA with deposition No. ECACC 96112640, and an applicable excipient. What is also presented is a method for the use of the pharmaceutical composition in the form of injections, as well as the use of a monoclonal antibody which recognises the CD6 human leukocyte differentiation antigen, for preparing a therapeutic agent.

EFFECT: group of inventions provides a prolonged therapeutic effect in the patients suffering rheumatoid arthritis after a short period of administration.

3 cl, 3 dwg, 3 ex

FIELD: medicine.

SUBSTANCE: bacterial Lactobacillus casei CNNM 1-1518 strain is used for preparing an oral composition for increasing humoral immunity induced in influenza vaccination, preferentially in patients aged 65 years and older. The composition is found in the form of an enzyme milk product, in the form of a foodstuff or a food additive, and with the content Lactobacillus casei CNNM 1-1518, preferentially 1x108 to 1x109 CFU/ml.

EFFECT: intensified humoral response induced by influenza vaccination.

10 dwg, 1 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine, specifically pharmacology, and may be used for correction of immune disorders in pathological conditions associated with Th1-dependent type of immune response (chronic, smoldering and recurrent infectious, as well as oncological diseases).

EFFECT: water-soluble polysaccharides of molecular weight 310 and 490 kDa recovered from compendial raw calendula are macrophage inflammatory properties activators, and Th-dependent type of immune response, and may extend the range of herbal products able to stimulate immune response in infectious-inflammatory processes and oncological disease.

2 ex, 4 tbl

Liquid formulations // 2470631

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to medicine and describes a pharmaceutical concentration for dissolution before oral introduction, containing S1P receptor modulator or agonist specified in a group involving 2-amino-2-[2-(4-octylphenylethyl)]propane-1,3-diol, 2-amino-2-[4-(benzyloxyphenylthio)-2-chlorophenyl]ethyl-1,3-propanediol or proper phosphate, and 1-{4-[1-(4-cyclohexyl-3-trifluoromethylbenzyloxyimino)ethyl]-2-ethylbenzyl}azetidine-3-carboxylic acid, or their pharmaceutically acceptable salts, respectively, and 65 to 99 wt % propylene glycol, and optionally one or more other solvents, one or more aromatisers and/or one or more preserving agents; all the ingredients are added up to 100 wt %. What is also described is a pharmaceutical solution containing the concentrate, the use of the concentrate and the method for treating an individual in need of immune system suppression with the use of said concentrate.

EFFECT: invention provides physical stability over long periods of time.

9 cl, 6 ex

FIELD: chemistry.

SUBSTANCE: invention relates to biologically active peptide complexes with immunomodulating and antiviral activity. The disclosed peptide complexes have a three-dimensional structure wherein X1 is absent or contains at least 1 amino acid; R1 and R2 are peptide chains which contain amino acid residues, His or Cys, capable of reacting with transition metal ions, wherein R1 contains up to 5 amino acid residues or is absent; R2 contains up to 3 amino acid residues or is absent.

EFFECT: peptide complexes rich in histidine and, primarily alloferon peptides with Zn ions, enable to produce preparations with a directed mechanism of action and enable their design in accordance with the understanding of the structure of the medicinal target.

3 cl, 7 dwg, 2 tbl, 6 ex

FIELD: medicine.

SUBSTANCE: complex therapy, including introduction of antibiotics, is performed. Additionally carried out is immunomodulating therapy 2 ml of 0.2% superlimf during and for 5-7 days after operation 2 times per day.

EFFECT: reduction of treatment terms due to normalisation of functional state of immune and antioxidant systems in said category of patients.

7 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to medicine and is intended for prevention of hepatobiliary dysfunctions in children, living in conditions of contamination of atmospheric air with phenol, formaldehyde, methanol. Complex of medications ia applied. Eslidin in introduced perorally in 14-day course in age dosage from 3 to 7 years - 1 capsule 2 times per day, over 7 years - 1 capsule 3 times per day. Canephron N is introduced perorally, diluted in small amount of water, 2 times per day, to children of pre-school age in dose 10 drops, of school age - 20 drops per day with 10 day course; Flamin is introduced perorally 30 minutes before meal in dose 1-2 tablets per day with 10 day course; Jungle is introduced perorally during or after meal in dose 1 tablet 1 time per day with 21 day course; Immunal is introduced perorallyt 1 time per day with 10 day course in age dosage from 1 year to 6 years - 5-10 drops, from 6 to 12 years - 10-15 drops, over 12 years - in dose 20 drops. Course is repeated 1 time per year in children with light degree of disease course and 2 times per year in case of middle-severe disorders.

EFFECT: method makes it possible to ensure and prolong remission and reduce frequency of recurrences of hepatobiliary dysfunctions in children.

10 tbl, 1 ex

Up!