Docosahexaenoic acid derivatives and their application as drugs


FIELD: medicine.

SUBSTANCE: invention refers to new compounds of formula (I) where X is carboxylic acid, carboxylates, carboxylic anhydride, diglyceride, triglyceride, phospholipid, or carboxamides, or to any their pharmaceutically acceptable salt. The invention particularly refers to (4Z, 7Z, 10Z, 13Z, 16Z, 19Z)-ethyl 2-ethyldocosa-4,7,10,13,16,19-hexanoate. The invention also refers to a food lipid composition and to a composition for diabetes, for reducing insulin, blood glucose, plasma triglyceride, for dislipidemia, for reducing blood cholesterol, body weight and for peripheral insulin resistance, including such compounds. Besides, the invention refers to methods for treating and/or preventing diabetes, dislipidemia, peripheral insulin resistance, body weight reduction and/or weight gain prevention, insulin, blood cholesterol, blood glucose and/or plasma triglyceride reduction.

EFFECT: higher clinical effectiveness.

61 cl, 4 tbl, 16 dwg, 5 ex

 

The technical FIELD TO WHICH the PRESENT INVENTION

The invention relates to compounds of General formula (I):

and their use as pharmaceuticals, in particular for the treatment of diabetes of the 2nd type and pre-diabetic States. Also the present invention relates to pharmaceutical compositions comprising the compounds of formula (I), and to the composition of fatty acids, including the compounds of formula (I).

Background FOR the PRESENT INVENTION

The increasing incidence of diabetes of the 2nd type in the entire world is a huge medical problem and puts the need of implementing a successful preventive and therapeutic measures. The increasing incidence of obesity and overweight, which are strongly correlated with diabetes of the 2nd type, the negative impact on the treatment of diabetes and increases the risk of diseases associated with hypertension, dyslipidemia and atherosclerosis.

Pathophysiological condition that precedes the development of type 2 diabetes-type is associated with a reduced effect of insulin on peripheral tissues, this phenomenon is called insulin resistance. These fabrics are mainly muscle, adipose tissue and liver tissue. When type 2 diabetes type insulin resistance most charactername muscle tissue. Syndrome, in which the observed insulin resistance, elevated blood pressure, dyslipidemia and inflammatory condition called metabolic syndrome. The prevalence of metabolic syndrome in the adult population in developed countries is 22-39% (Meighs 2003).

Currently, the most promising way to reduce metabolic syndrome is a change in lifestyle, which consists in reducing weight, reducing the consumption of saturated fat, increasing physical activity, in combination with appropriate pharmacotherapy. When healthy diets that allow you to avoid unnecessary energy consumption, saturated fats are replaced with mono - and polyunsaturated fatty acids. In the prevention of type 2 diabetes type especially its effectiveness has proven omega-3 polyunsaturated fatty acids found in oily fish, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA).

EPA and DHA affect various physiological processes such as regulation of lipid levels in plasma, cardiovascular and immune function, insulin action, development of the nervous system and visual function. There is evidence of the role of these acids in the prevention and correction of ischemic heart disease, dyslipidemia, type 2 diabetes type insulin resistance is hypertension (Simonopoulos 1999; Geleijnse 2002; Storlien 1998). Recent studies have shown that omega-3 polyunsaturated fatty acids are important mediators of gene expression, acting through nuclear receptors, such as receptors, activating peroxisome proliferation (PPARs), controlling the expression of genes related to carbohydrate and lipid metabolism and lipogenesis (Jump 2002). Receptors PPARs are nuclear receptors that play an important role in the development of obesity-related metabolic disorders such as hyperlipidemia, insulin resistance and coronary heart disease.

Three subtypes of receptors, α, γ and δ, have different expression patterns and recognize various components of lipoproteins, and regulate lipid homeostasis based on the needs of specific tissues. PPARα increases the catabolism of fatty acids in the liver and is the molecular target of fibrates to reduce the level of lipids. PPARγ plays a role in the differentiation of adipocytes and mediates the activity of insulin-sensitizing agents - preparations of thiazolidinediones (glitazones), the mechanisms of this mediation are poorly understood (Chih-Nao 2003; Yki-Jarvinen 2004).

Recently, drugs that act as ligands of PPARγ receptor, were used in the treatment of type 2 diabetes type (Yki-Jarvinen 2004). These compounds, called preparations of thiazolidinediones or glitazones are drugs, to the verge reversiruyut insulin resistance, which is the pathophysiological basis for the development of metabolic syndrome and type 2 diabetes type. These compounds, of which rosiglitazone and pioglitazone are available as medicines to reduce the concentration of glucose in fasting and postprandial glucose concentration (as was shown with the help of test glucose tolerance (IGT), insulin levels in plasma and the concentration of free fatty acids. In this respect, glitazone act as insulin sensitizers.

However, these improvements are usually accompanied by weight gain and increased mass of adipose tissue (Adams 1997). The use of preparations of thiazolidinediones is not only associated with weight gain, but some patients also experience a delay of a liquid and increase of plasma volume, resulting in peripheral edema. The increase of body weight and edema can lead to cardiac activity, this was the reason that the Management under the control over products and medicines (FDA) has included the warning information rosiglitazone (Avandia) and pioglitazone (Takeda). These adverse effects limit the use of glitazones especially in patients with ischemic heart disease. Clearly, there is a need to develop new drugs that have a positive impact on resistance to what Sulina, but reduce the body weight and do not contribute to water retention in the body.

Effect of polyunsaturated fatty acids (PUFAs) on receptors PPARs is not only the result of the structure of fatty acids and affinity to the receptor. Also important factors affecting the concentration of nonesterified fatty acids (NEFA). On NEFA pool is affected by the concentration of exogenous fatty acids coming into the cell, and the number of endogenously synthesized fatty acids, their removal through incorporation into lipids, and their way of oxidation (Pawar 2003).

Although omega-3 fatty acids are weak agonists of PPARs in comparison with pharmacological agonists as tioglitazone, these fatty acids showed improved glucose and increase insulin sensitivity (Storlien 1987). Report was made that the adipocytes were more sensitive to insulin and transported more glucose when the diet was increased, the ratio of polyunsaturated fatty acids to saturated fatty acids (Field 1990). Together, these data indicate that the C20 and C22 fatty acids called EPA and DHA, may play a role in preventing the development of insulin resistance.

Because of their limited stability in vivo and the absence of biological specificity PUFAs are not really widely used therapeutic agents. Several of the research groups were performed chemical modification of n-3 polyunsaturated fatty acids in order to change or improve their metabolic effects.

For example, lipid-lowering effect of EPA was reinforced by the introduction of methyl or ethyl in the α - or β-position EPA (Vaagenes 1999). These compounds also reduce the level of free fatty acids in plasma, whereas ethyl ester EPA has no such effect.

In recent work published by L. Larsen (Larsen 2005), the authors show that the α-methyl derivatives of EPA and DHA increase the activation of the nuclear receptor PPARα and thus the expression of L-FABP in comparison with EPA/DHA. EPA with the ethyl group in α-position activates PPARα with the same strength as α-methyl-EPA. The authors suggest that a slow catabolism of these α-methyl fatty acids may contribute to their increased effects due to reduced β-oxidation in mitochondria, leading to paroksizmalnom oxidation.

It was shown that alpha-methyl-EPA is a stronger inhibitor of platelet aggregation than EPA, both in vitro (Larsen 1998)and in vivo (Willumsen 1998). Abstract of Japan patent, publication number 05-00974, reported DHA, substituted in the alpha position of the Oh-group, but only as an intermediate product. About the research potential of the pharmaceutical effects of this compound has not been reported.

The company Laxdale Limited also described the use of alpha-substituted derivatives of EPA in the treatment of psychiatric disorders and disorders of the Central nervous system (US 6689812).

None of these modified fatty acids were not, however, satisfactory pharmaceutical activity and none of them appeared on the pharmaceutical market.

SUMMARY of the INVENTION

The aim of the present invention to provide new derivatives of DHA, which have a therapeutic effect.

On the basis of the present invention presents a number of aspects in the attached claims. Some of these aspects are the following aspects:

1. New connections, i.e. some α-substituted derivatives of polyunsaturated fatty acids.

2. New compounds for use as drugs and for use in therapy.

3. Composition of fatty acids or pharmaceutical composition that includes the new connection.

4. Composition of fatty acids, including new compounds for use as drugs and for use in therapy.

5. The use of new compounds for obtaining a medicinal product for the prevention and/or treatment of diabetes in humans or in animals.

6. The use of new compounds for obtaining a medicinal product for the treatment and/or prevention of obesity or overweight.

7. The use of new compounds for obtaining medicines descontrola reduction of body weight and/or to prevent gaining weight.

8. The use of new compounds for obtaining a medicinal product for the treatment and/or prevention of diseases associated with amyloidosis.

9. The use of new compounds for obtaining a medicinal product for the treatment or prophylaxis of multiple risk factors for cardiovascular diseases.

10. The use of new compounds to obtain drugs for prevention of cerebral circulation, cerebral or transient ischaemic attacks related to atherosclerosis of certain arteries.

11. Method specific treatment of diabetic diseases, preferably of type 2 diabetes type.

12. Method of controlling the loss of body weight, prevent gaining weight and/or treatment and/or prevention of obesity or overweight.

13. The method of treatment and/or prevention of diseases associated with amyloidosis

14. The method of treatment or prophylaxis of multiple risk factors for cardiovascular diseases.

15. The way to prevent violations of cerebral circulation, cerebral or transient ischaemic attacks related to atherosclerosis of certain arteries.

16. Methods of obtaining new analogues of fatty acids of the present invention

The present invention relates to the compound of formula (I):

where - R1and R2are the same or different and may be selected from the group consisting of hydrogen atom, hydroxyl group, alkyl group, halogen atom, alkoxygroup, alloctype, acyl group, alkenylphenol group, alkenylphenol group, aryl group, allylthiourea, alkoxycarbonyl group, alkylsulfonyl group, alkylsulfonyl group, amino group, alkylamino; and

- X means carboxyl group, carboxylate group, or carboxamido group or any pharmaceutically acceptable salt, MES, complex or prodrug, provided that:

the compound of formula (I) is not (fully-Z)-4,7,10,13,16,19-docosahexaenoic acid (DHA), alpha-methyl DHA, methyl ester, alpha-methyl DHA, acylovir ether, alpha-methyl DHA or ethyl ester of alpha-hydroxy-DHA.

Conditions apply to the following cases:

when R1is a hydrogen atom, R2is not a hydrogen atom;

when R2is a hydrogen atom, R1is not a hydrogen atom;

when R1is a methyl group, R2is not a hydrogen atom and X is a carboxyl group, methylcarbazole or ethylcarboxylate;

when R2is a methyl group, R1is not a hydrogen atom, and X is not carboxyl the Noah group, methylcarbazole or ethylcarboxylate;

when R1is a hydroxyl group, R2is not a hydrogen atom and X is not ethylcarboxylate; and

when R2is a hydroxyl group, R1is not a hydrogen atom and X is not ethylcarboxylate.

In connection corresponding to the present invention, the mentioned alkyl group may be selected from the group consisting of methyl, ethyl, n-sawn, ISO-propyl, n-butilkoi, second-butilkoi, n-hexylene and benzyl groups; the specified halogen atom may be selected from the group consisting of fluorine, chlorine, bromine and iodine; this alkoxygroup can be selected from the group consisting of methoxy, ethoxy-, propoxy-, isopropoxy-, second -, butoxy, phenoxy-, benzyloxy-, och2CF3and co2CH2Och3groups; this alloctype may be selected from acetoxy, propionoxy and butirosin; specified Alchemilla group can be selected from the group consisting of allyl, 2-butanediol and 3-hextile groups; specified Alchemilla group can be selected from the group consisting of propargyl, 2-botinelli and 3-hextile groups; mentioned aryl group is a phenyl group; this allylthiourea can be selected from the group consisting of methylthio, atilt the o-, isopropylthio and paneltop; specified alkoxycarbonyl group can be selected from the group consisting of methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl and butoxycarbonyl groups; specified alkylsulfonyl group can be selected from the group consisting of methanesulfonyl, ethanolamines and isopropanolamines groups; specified alkylsulfonyl group can be selected from the group consisting of methanesulfonyl, acanaloniidae and isopropanolamines groups; this alkylamino can be selected from the group consisting of methylamino-, dimethylamino-, ethylamino and diethylamino groups; specified carboxylate group can be selected from the group consisting of ethylcarboxylate, methylcarbamate, n-propellerblade, isopropylcarbonate, n-butylcarbamoyl, sec-butylcarbamoyl and n-lexiscanlexiscan; specified carboxamidine group can be selected from the group consisting of primary carboxamides, N-methylcarbamate, N,N-dimethylcarbamate, N-ethylcarbodiimide and N,N-diethylcarbamoyl groups.

In one embodiment of the present invention R1and R2selected from the group consisting of a hydrogen atom, hydroxy-group, alkyl group, halogen atom, alkoxygroup, allylthiourea, alkylsulfonyl GRU is dust, alkylsulfonyl group, amino group and alkylamino.

In another embodiment, the present invention R1and R2selected from the group consisting of a hydrogen atom, hydroxy-group, C1-C7alkyl group, halogen atom, a C1-C7alkoxygroup, C1-C7allylthiourea, C1-C7alkylsulfonyl group, C1-C7alkylsulfonyl group, amino group and C1-C7alkylamino. The specified C1-C7the alkyl group can be methyl, ethyl or benzyl group; specified halogen atom may be fluorine or iodine: the specified C1-C7alkoxygroup can be methoxy or ethoxypropane; the specified C1-C7allylthiourea may be methylthio, ethylthio or phenylthiourea; the specified C1-C7alkylsulfonyl group can be ethanolamines group; the specified C1-C7alkylsulfonyl group can be acanaloniidae group; the specified C1-C7alkylamino can be ethylamino or diethylaminopropyl; and X may mean ethylcarboxylate or carboxamido group.

In another embodiment, the present invention R1and R2selected from the group consisting of a hydrogen atom, a C2-C7alkyl group, an atom Gal is gene C1-C7alkoxygroup, C1-C7allylthiourea, C1-C7alkylsulfonyl group, C1-C7alkylsulfonyl group, amino group and C1-C7alkylamino; and X is a carboxylate. Specified With2-C7the alkyl group may be ethyl or benzyl group; specified halogen atom may be fluorine or iodine; the specified C1-C7alkoxygroup can be methoxy or ethoxypropane; the specified C1-C7allylthiourea may be methylthio, ethylthio or phenylthiourea; the specified C1-C7alkylsulfonyl group can be ethanolamines group; the specified C1-C7alkylsulfonyl group can be acanaloniidae group; the specified C1-C7alkylamino can be ethylamino or diethylaminopropyl; and X can mean ethylcarboxylate.

In the compound corresponding to formula (I) of the present invention, R1and R2may be the same or different. When they are different, the compounds of formula (I) can exist in stereoisomeric forms. It is understood therefore that this invention includes all optical isomers of compounds of formula (I) and mixtures thereof, including racemates.

Consequently, where R1differs from R2, the present invention includes soybean is inane formula (I), which is racemic or enantiomerically pure, or as (S) or (R) enantiomer. Consequently, where R1differs from R2, the present invention includes compounds of formula (I), which are racemic or enantiomerically pure, or as (S) or (R) stereoisomer.

Within the present invention are the enantiomers of the compounds of formula (I)as defined above. In addition, the enantiomers of derivatives of DHA of the present invention may be in the form of a carboxylic acid or its pharmaceutically acceptable salt, any ester, anhydride or amide (primary, secondary, tertiary). Derived acid may be in the form of a phospholipid, or tri-, di - or monoglyceride.

In one embodiment, the compounds of formula (I)corresponding to the present invention, one of R1and R2means2-C7alkyl group, for example, ethyl or benzyl, and the other denotes a hydrogen atom. Preferably, the alkyl group is ethyl.

In another embodiment, compounds of formula (I)corresponding to the present invention, one of R1and R2means alkoxygroup, for example, ethoxy - or methoxy group and the other denotes a hydrogen atom.

In another embodiment, compounds of formula (I)corresponding to this izobreteny is, one of R1and R2means a halogen atom, for example, fluorine or iodine, and the other denotes a hydrogen atom.

In another embodiment, compounds of formula (I)corresponding to the present invention, one of R1and R2means allylthiourea, for example, ethylthio-, methyltin or phenylthiourea, and another mean a hydrogen atom. Preferably allylthiourea is ethylthiourea.

In another embodiment, compounds of formula (I)corresponding to the present invention, one of R1and R2means alkylsulfonyl group, for example ethylsulfonyl, and another mean a hydrogen atom.

In another embodiment, compounds of formula (I)corresponding to the present invention, one of R1and R2means the amino group and the other denotes a hydrogen atom.

In another embodiment, compounds of formula (I)corresponding to the present invention, one of R1and R2means alkylamino, for example, ethylamino or diethylaminopropyl, and another mean a hydrogen atom.

In a further embodiment, compounds of formula (I)corresponding to the present invention, R1and R2are the same and mean C1-C7is an alkyl group, preferably a methyl or ethyl group.

In the preferred VA is Ianto implementation of the compounds of formula (I), X represents a carboxylate, for example, ethylcarboxylate.

The connection corresponding to the present invention can exist in the form of a phospholipid, three-, di - or monoglyceride or in the form of the free acid.

Alpha-substituted derivatives of DHA, corresponding to the present invention, pharmaceutically were very active. In particular, derivatives of fatty acids corresponding to the present invention, have great potential for use in the treatment and/or prevention of diabetes and pre-diabetic States.

Another aspect of the present invention relates to the compound of formula (I) for use as a medicine.

The present invention also relates to a method for obtaining compounds of formula (I). For example, the compound of formula (I) can be obtained from the (fully-Z)-4,7,10,13,16,19-docosahexaenoic acid (DHA). DHA can be of vegetable, microbial and/or animal origin (for example, fat sea fish). Another important advantage of the compounds of formula (I) is that the analogues of fatty acids can be obtained directly on the basis of (fully-Z)-4,7,10,13,16,19-docosahexaenoic acid (DHA).

In a preferred embodiment of the present invention analogues of fatty acids of the formula (I) obtained on the basis of DHA, where specified DHA obtained at least from the underwater source of vegetable, microbial and animal origin or their combination. The present invention includes, therefore, derivative, obtained from fat microbial origin containing DHA. Specified DHA derived from marine oil, such as fish oil.

Another aspect of the present invention relates to a pharmaceutical composition comprising a compound of formula (I) as an active ingredient. The pharmaceutical composition may include a pharmaceutically active carrier. Accordingly, the pharmaceutical composition corresponding to the present invention, designed for oral administration, for example, in the form of a capsule or sachet powder. The appropriate daily dosage of the compounds of formula (I)corresponding to the present invention is from 10 mg to 10 g, in particular from 100 mg to 1 g of the specified connection.

In addition, the present invention relates to the composition of fatty acids comprising the compound of formula (I). At least 60% or at least 90% of the weight of the composition of fatty acids can make the specified connection. Composition of fatty acids may optionally include (fully-Z)-5,8,11,14,17-eykozapentaenovuyu acid (EPA) (an-2)-4,7,10,13,16,19-docosahexaenoic acid (DHA),-Z)-6,9,12,15,18-kanakesanturai acid (NDA)and/or (fully-Z)-7,10,13,16,19-docosapentaenoic acid (DPA). Girn the e acid may be present in the form of derivatives. Composition of fatty acids corresponding to the present invention may optionally include pharmaceutically acceptable antioxidant, such as tocopherol. Within the present invention is also the composition of the fatty acids described above, for use as pharmaceuticals.

In an additional aspect, the present invention relates to the use of compounds corresponding to formula (I), for the manufacture of a medicinal product for controlling body weight reduction and/or prevention of gaining weight; for the manufacture of a medicinal product for the treatment and/or prevention of obesity or overweight; for the manufacture of a medicinal product for the prevention and/or treatment of diabetes in animals, especially type 2 diabetes type; for the manufacture of a medicinal product for the treatment and/or prevention of diseases associated with amyloidosis; for the manufacture of a medicinal product for the treatment or prophylaxis of multiple risk factors for cardiovascular diseases, especially for the treatment of elevated lipid levels in the blood, for the manufacture of a medicinal product for the prevention of violations of cerebral circulation, cerebral or transient ischaemic attacks related to atherosclerosis of certain arteries.

In addition, the present invention relates to sposobu control body weight reduction and/or prevention of gaining weight; the method of treatment and/or prevention of obesity or overweight; the method of prevention and/or treatment of diabetes, especially type 2 diabetes type; the method of treatment and/or prevention of diseases associated with amyloidosis; the method of treatment or prophylaxis of multiple risk factors for cardiovascular disease; the way to prevent violations of cerebral circulation, cerebral or transient ischaemic attacks related to atherosclerosis of certain arteries, where the pharmaceutically effective amount of the compounds of formula (I) is administered to a human or animal. Accordingly, the compound of formula (I) is used orally in humans or animals.

BRIEF DESCRIPTION of DRAWINGS

Figure 1 is a schematic description of theory of the pool of free fatty acids.

The figure 2 describes the models and methods used in the present invention to demonstrate the effects on the metabolic syndrome and type 2 diabetes type.

The figure 3 shows the concentration of free fatty acids of various compounds of the present invention, in the liver tissue of animals treated with these compounds at a concentration of 1.5% of the total fat content.

The figure 4 shows the intracellular concentration of DHA in the liver tissue of animals receiving different compounds corresponding to this is th invention, at a concentration of 1.5% of the total fat content.

The figure 5 shows the affinity of binding to receptor PPARγ various compounds of the present invention.

The figure 6 shows the affinity of binding to receptor PPARα various compounds of the present invention.

The figure 7 shows the affinity of binding to receptor RXRα various compounds of the present invention.

The figure 8 shows the release of luciferase transfected cells treated with various compounds disclosed in this invention,

The figure 9 shows the plan of the experiment unit 4.

The figure 10 shows the change of body weight within 2 weeks of experimental diet after 8 weeks of a diet high in fat.

The figure 11 shows the results of luciferase activity, i.e. endogenous PPARγ activity.

The figure 12 shows the endogenous luciferase activity in various compounds of the present invention, compared with DHA.

The figure 13 shows a typical curve of decrease in blood glucose levels before and after animals with insulin resistance was given connection, reducing insulin resistance.

In figures 14, 15 and 16 shows the different effects derived DHA corresponding to this is th invention, on the metabolic syndrome and insulin resistance.

DETAILED description of the INVENTION

In the research work leading to the present invention were obtained new derivatives of DHA, which showed high pharmaceutical activity.

Fatty acids penetrate the cell passively or through G-protein conjugate transport systems, such as transport proteins fatty acids. Inside the cells they temporarily bind binding proteins (proteins, fatty acid binding, FABP), which play an important role in the direction of fatty acids to the different cells for metabolism and gene expression (Pawar & Jump 2003). (Figure 1 cell of the liver).

Esterification fatty acids in triglycerides, polar lipids, cholesterol esters and beta-oxidation (mitochondrial and peroxisomal) requires the conversion of fatty acids to acyl COA the thioethers. Other routes of metabolism, as microsomal NADPH-dependent minoocycline and the synthesis of eicosanoids use free fatty acids as substrates. All of these reactions, apparently, affect the concentration in the cell free fatty acids (nonesterified) and, thus, on the number and type of fatty acids that can be used as ligands to nuclear receptors. Since it is known that receptors PPARs tie the season with free fatty acids, it seems a fair assumption that the composition of the pool of fatty acids is an important factor in the activity of PPAR.

On the composition of the pool of free fatty acids is affected by the concentration of exogenous fatty acids entering the cells, and the rate of their removal through metabolism pathways listed above. Because fatty acids with short and medium chains are required for these pathways of metabolism, practically only polyunsaturated fatty acids with long chains will be free to bind to nuclear receptors. In addition, an important factor may also be the structure of the fatty acids. Even if a series of mono - and polyunsaturated fatty acids showed an affinity for the receptor PPARα, EPA and DHA showed the highest binding ability in experiments with cells rat liver (Pawar & Jump 2003).

Search candidates fatty acids available for genetic modification of proteins by interacting with nuclear receptors, such PPARs, important to acknowledge that these fatty acids will enrich the pool of free fatty acids.

DHA, which is included in the cell, quickly transformed into acyl-COA the thioethers and incorporated into phospholipids, resulting in the intracellular concentration of DHA is relatively low. These DHA-COA is also a substrate for β-oxidation initially peroxisome that mean is it for retroconversion of DHA EPA see figure 1. Due to the rapid incorporation into neutral lipids and oxidation of DHA will not linger long in the pool of free fatty acids. As a consequence, the effect of DHA on gene expression may be limited.

The present invention aims rather the accumulation of fatty acids derivatives in the pool of free fatty acids than incorporation into phospholipids. The inventors surprisingly found that the introduction of at least one of the substituent in α-position DHA results in a lower oxidation rate and a lower incorporation into neutral lipids. This leads to increased influence on gene expression, as derived DHA will accumulate in the liver tissue, muscle tissue, fat cells and stimulate the activity of nuclear receptors to a greater extent than DHA.

Different substituents corresponding to the present invention, will give varying degrees of affinity derivatives to receptors. It is also possible that changes in affinity for proteins, fatty acid binding, lead to changes in biological activity of these α-substituted derivatives of DHA formula (I). Overall, these changes result in improved therapeutic effect derived DHA of the present invention, compared with DHA.

EPA (fully-Z)-5,8,11,14,17-eicosapentaenoic acid) was previously alkylated at the α - and β-position with the purpose of the inhibition of mitochondrial β-position. DHA is not oxidized in the mitochondria, and is included in the phospholipids. In peroxisome certain amount of DHA reconvertible in EPA. The substituent in the α-position of EPA and DHA will affect various metabolic pathways. Previously it was shown that α-methyl-EPA and β-methyl-EPA included in phospholipids and triglycerides, and α-ethyl-EPA no (Larsen 1998). In this study derivatives were tested as substrates and/or inhibitors of enzymes that play a role in the cascade of eicosanoids. Since most of the substrates for these enzymes is a fatty acid released from phospholipids, was desired in order derivatives were incorporated into phospholipids. In contrast, as mentioned above, we wanted to derivatives not included in lipids and accumulated in the NEFA pool.

In this description, the abbreviation "PRB-x", where x is an integer, will be used in the description of specific compounds of the present invention. Listed below are the structural formulas and common names of these compounds.

PRB-1 ethyl ester of α-methylacetoacetate acid

PRB-2 ethyl ester of α-acidogenicity acid

PRB-3 ethyl ester of α-toxicologically acid

PRB-ethyl 4 the ether α-forodolcegusto acid

PRB-5 ethyl ester of α,α-dimethylpolysiloxene acid

PRB-6 ethyl ester of α-dimethylpolysiloxene acid

PRB-7 ethyl ester of α-diethyldithiocarbamic acid

PRB-8 ethyl ester of α,α-diethyldodecanamide acid

PRB-9 ethyl ester of α-benzylideneacetone acid

PRB-10 ethyl ester of α-econsultantresources.com acid

PRB-11 ethyl ester of α-titaniloksalata acid

PRB-12 ethyl ester of α-hydroxydocosahexaenoic acid

PRB-13 amide α-methylacetoacetate acid

PRB-14 ethyl ester of α-methoxyacetophenone acid

PRB-15 ethyl ester of α-iodocholesterol acid

PRB-17 ethyl ester of α-aminomonosaccharide acid

PRB-18 (4R,5S)-3-docosahexaenoyl-4-methyl-5-phenyl-oxazolidin-2-he

PRB-19 (4R,5S)-3-[(S)-α-acidogenicity]-4-methyl-5-phenyl-oxazolidin-2-he

PRB-20 ethyl ester of (S)-(+)-α-acidogenicity acid

PRB-21 (4S,5R)-3-docosahexaenoyl-4-methyl-5-phenyl-oxazolidin-2-he

PRB-22 (4S,5R)-3-[(R)-α-acidogenicity]-4-methyl-5-phenyl-oxazolidin-2-he

PRB-23 ethyl ester (R)-(-)-α-acidogenicity acid

PRB-24 ethyl ester of 2-(1,3-Dioxo-1,3-dihydro-isoindole-2-yl)-docosahexaenoic acid

PRB-25 ethyl ester of α-ethylaminoethanol acid

PRB-26 ethyl ester of α-diethylaminotoluene acid

PRB-1 corresponds to the compound of formula (I), where R1or R2means methyl and second means hydrogen, and X means ethylcarboxylate.

PRB-2 corresponds to the compound of formula (I), where R1or R2means ethyl and second means hydrogen, and X means ethylcarboxylate.

PRB-3 corresponds to the compound of formula (I), where R1or R2means ethoxypropan and second means hydrogen, and X means ethylcarboxylate.

PRB-4 corresponds to the compound of formula (I), where R1or R2means fluorine, and the other is hydrogen, and X means ethylcarboxylate.

PRB-5 corresponds to the compound of formula (I), where R1and R2means ethyl, and X means ethylcarboxylate.

PRB-6 corresponds to the compound of formula (I), where R1or R2means metalcorp, and X means ethylcarboxylate.

PRB-7 corresponds to the compound of formula (I), where R1or R2means ethylthiourea and the other is hydrogen, and X means ethylcarboxylate.

PRB-8 corresponds to the compound of formula (I), where R1and R2means ethyl and the other is hydrogen, and X means ethylcarboxylate.

PRB-9 corresponds to the compound of formula (I), where R1or R2means benzyl and the other is hydrogen, and X means ethylcarboxylate.

PRB-10 corresponds to the compound of formula (I), where R1or R2means econsulting and the other is hydrogen, and X means ethylcarboxylate.

PRB-11 corresponds to the compound of formula (I), where R1or R2means phenylthiourea and the other is hydrogen, and X means ethylcarboxylate.

PRB-12 corresponds to the compound of formula (I), where R1or R2means a hydroxy-group and the other is hydrogen, and X means ethylcarboxylate.

PRB-13 corresponds to the compound of formula (I), where R1or R2means methyl and the other is hydrogen, and x is the primary carboxamide.

PRB-14 corresponds to the compound of formula (I), where R1or R2means a methoxy group and dragojevica hydrogen, and X means ethylcarboxylate.

PRB-15 corresponds to the compound of formula (I), where R1or R2is iodine and the other is hydrogen, and X means ethylcarboxylate.

PRB-17 corresponds to the compound of formula (I), where R1or R2means the amino group and the other is hydrogen, and X means ethylcarboxylate.

PRB-20 corresponds to the (S) stereoisomer of the compounds of formula (I), where R1or R2means ethyl and the other is hydrogen, and X means ethylcarboxylate.

PRB-23 corresponds to the (R) stereoisomer of the compounds of formula (I), where R1or R2means ethyl and the other is hydrogen, and X means ethylcarboxylate.

PRB-24 corresponds to the compound of formula (I), where R1or R2means N-phthalimide and the other is hydrogen, and X means ethylcarboxylate.

PRB-25 corresponds to the compound of formula (I), where R1or R2means ethylamino and the other is hydrogen, and x is the primary carboxamide.

PRB-26 corresponds to the compound of formula (I), where R1or R2means diethylaminopropyl and the other is hydrogen, and X means ethylcarboxylate.

PRB-2 is the most preferred compound corresponding to the present invention. Other preferred compounds disclosed in this invention are PRB-5, PR-7 and PRB-8.

It should be understood that the present invention includes any pharmaceutically acceptable salt, solvate, complexes or prodrugs of the compounds of formula (I).

"Prodrugs" are substances, which may or may not possess pharmacological activity as such, but may be introduced into the body (e.g., oral or parenteral), where they will be exposed to bioactivate (for example, metabolized), which will result in the received agent of the present invention, which is pharmacologically active.

Where X is a carboxylic acid, the present invention also includes salts of carboxylic acids. The corresponding pharmacologically acceptable salts carboxypropyl include metal salts, such as, for example aluminium, alkali metal salts such as lithium, sodium or potassium, salts of alkaline earth metals such as calcium, magnesium and ammonium salts and substituted ammonium salts.

"Therapeutically effective amount" means an amount of therapeutic agent, which gives the expected result. As individual needs may vary, determination of optimal amounts of each oxide adduct of nitrite is within the competence of a person skilled in the art. As a rule, the scheme applied for the treatment of illness is any and violations compounds and/or compositions of the present invention is selected in accordance with a variety of factors, including the age, weight, sex, diet, and General condition of the patient.

Under "drug" means a compound corresponding to the formula (I), in any form, suitable for use for medical purposes, for example in the form of a medicinal product, pharmaceutical product, or product, diet product, food product or food additive.

In the context of this specification, the term "therapy" also includes "prevention", unless there are specific indications to the contrary. Similarly apply the terms "therapeutic" and "therapeutically".

Treatment includes any therapeutic application, which can be useful for a person or animal. The treatment preferably mammals. As the treatment of the person, and the treatment of animals is within the present invention. Treatment may be the treatment of an existing pathological condition or it can be preventive. The treatment may be carried out in relation to human adults, juvenile subjects, children, fetuses, or parts (e.g., organ, tissue, cell or molecule of nucleic acid). Under "chronic treatment" means treatment that lasts for weeks or years.

The term "therapeutically or pharmaceutically active amount" means the amount which the cut is ltate gives the desired pharmacological and/or therapeutic effects. The connection corresponding to the present invention, may, for example, to be included in the food product, dietary Supplement, nutritional Supplement or diet product. Alpha-substituted derivatives of DHA and EPA (or DHA) can be linked together or merged in triglyceride form by esterification between a mixture of alpha-derivatives, EPA and glycerol catalyzed using Novozym 435 (commercially available lipase from Candida antarctica immobilized form).

The compounds of formula (I) have activity as pharmaceuticals, in particular as triggers activity of nuclear receptors. Thus, the present invention also relates to compounds of formula (I), pharmaceutically acceptable salts, solvate, complex, or their prodrugs as defined above, for use as medicaments and/or for use in therapy. Preferably new connection, or a pharmaceutically acceptable salt, solvate, complex or prodrug of the present invention may be used:

- for the prevention and/or treatment of diabetes in humans and animals; for controlling body weight reduction and/or prevention of gaining weight;

- for the prevention and/or treatment of obesity or overweight in humans or in animals;

- for the treatment and/or prevention is IKI diseases, related amyloidosis;

- for the treatment or prophylaxis of multiple risk factors for cardiovascular disease;

- to prevent violations of cerebral circulation, cerebral or transient ischaemic attacks related to atherosclerosis of certain arteries.

- for the treatment of tuberculosis or HIV.

There are two forms of diabetes. One form is the diabetes of the 1st type, which is known as insulin-dependent diabetes mellitus (IDDM), and another form is type 2 diabetes type, which is known as insulin-independent diabetes mellitus (NDDDM). Diabetes of the 2nd type is associated with obesity/overweight and lack of exercise, the beginning of his often gradual, usually 2 diabetes type occurs in adults and is caused by reduced insulin sensitivity. This leads to a compensatory increase the production of insulin. This stage before the development of the diabetes of the 2nd type is called metabolic syndrome and is characterized by hyperinsulinemia, insulin resistance, obesity, impaired glucose tolerance, elevated blood lipids, hypercoagulopathy, dyslipidemia and inflammation, often leading to atherosclerosis of the arteries. Later, when disturbed secretion of insulin, develop diabetes of the 2nd type.

In a preferred embodiment, the connection with testwuide the formula (I), can be used for the treatment of diabetes of the 2nd type. Compounds corresponding to formula (I)may also be used to treat other types of diabetes selected from the group comprising metabolic syndrome, secondary diabetes, such as pancreatic, extrapancreatic/endocrine or diabetes, caused by the action of drugs, or special forms of diabetes, such as lipotropics, Metodicheskie or disease caused by a violation of insulin receptors. The present invention also includes the treatment of diabetes of the 2nd type. Accordingly, the compounds of formula (I)as defined above, can activate nuclear receptors, preferably PPAR (receptors, activating peroxisome proliferation) α and/or γ.

The compounds of formula (I) may also be used for treatment and/or prevention of obesity. Obesity is usually associated with increased insulin resistance, and obese people are at high risk of developing diabetes of the 2nd type, which is a major risk factor for cardiovascular disease. Obesity is a chronic disease that affects a growing number of people in Western countries, obesity is associated not only with the moral suffering, but also with decreased life span and numerous problems, for example, diabetes di is beta, insulin resistance and high blood pressure. Thus, the present invention meets a long-felt need in creating medicines that will decrease the overall weight of the body or the amount of adipose tissue in people suffering from obesity to lose weight before their ideal body weight and no significant unwanted side effects.

Compounds corresponding to formula (I)may also be used for the prevention and/or treatment of diseases associated with amyloidosis. Related amyloidosis pathological condition or disease associated with deposition of amyloid, preferably as a result of plaque formation include Alzheimer's disease or dementia, Parkinson's disease, amyotrophic lateral sclerosis, spongiforme encephalopathy, Krefeld-Jakob disease, cystic fibrosis, primary and secondary renal amyloidosis, IgA nephropathy, the amyloid deposits in the arteries, myocardium and nervous tissue. These diseases can be sporadic, inherited or even associated with infections such as tuberculosis or HIV, and often appear only at a later stage of life, even if non-hereditary forms can appear much earlier. Each disease is associated with a particular protein or a combination of these proteins, which are believed to cause patologica the fir States, associated with the disease. Treatment-related amyloidosis diseases can be either short-term (acute)and long-term (chronic).

The compounds of formula (I) can also be used for treatments directed at reducing amyloid accumulation, prevent protein misfolding, which can lead to the formation of so-called plaques, treatment aimed at reducing the production of the protein precursor, such as β-protein (beta-amyloid protein), and prevention and/or treatment aimed at inhibiting or slowing the formation of protein clusters or plaques. Warning tangles by using compounds of the formula (I)as defined above, is also included in the present invention. In one embodiment, the new compound, pharmaceutically acceptable salt, solvate, complex or prodrug, as defined above, are used to treat tuberculosis or HIV (human immunodeficiency virus).

Further, the compounds of formula (I) can be used in patients with symptoms of atherosclerosis of arteries supplying the brain, for example with ischemic stroke or transient ischemic attacks with the aim of reducing the risk of future, possibly fatal attack.

The compounds of formula (I) can also be used for the treatment of the Oia elevated levels of lipids in the blood.

In addition, the compounds of formula (I)as defined above, can be used for treatment and prophylaxis of multiple risk factors for cardiovascular diseases such as high blood pressure, hypertriglyceridemia and high coagulation activity of factor VII. Preferably the compounds of formula (I) used for the treatment of elevated blood lipids in humans.

The compounds of formula (I) and pharmaceutically acceptable salts, solvate prodrugs or complexes can be used by themselves, but usually will be introduced in the form of pharmaceutical compositions in which compounds of formula (I) (active ingredient) is associated with a pharmaceutically acceptable excipient, diluent or carrier.

The present invention thus also relates to pharmaceutical compositions comprising a therapeutically effective amount of the compounds of formula (I) of the present invention and a pharmaceutically acceptable carrier, solvent or excipient (including combinations thereof).

This pharmaceutical composition, which comprises or consists of a therapeutically effective amount of pharmaceutically active agent. The pharmaceutical composition preferably includes a pharmaceutically acceptable carrier, solvent or excipient (including combinations thereof). Applicable to the s carriers or diluents for therapeutic use are well known in the pharmaceutical field. The choice of a pharmaceutical carrier, excipient or solvent depends on the intended route of administration and standard pharmaceutical practice. The pharmaceutical composition can include (or additionally), the carrier, excipient or solvent, any suitable binder(s), lubricating substance(s), suspendisse substance(s), coating agent(s), solubilizing substance(s).

Pharmaceutical compositions within the present invention can include one or more of the following chemicals: preservatives, solubilizing agents, stabilizing agents, moisturizing agents, emulsifiers, sweeteners, colorants, flavouring agents, odorants (compounds of the present invention can be in the form of pharmaceutically acceptable salts, buffers, coating agents, antioxidants, suspendresume substances, adjuvants, fillers and solvents.

The pharmaceutical composition corresponding to the present invention, preferably formulated for oral administration to humans and animals. The pharmaceutical composition can also be prepared for any route of administration in which the active ingredients can be effectively absorbed and used, for example, intravenously, subcutaneously, intramuscularly, and tramazoline, rectally, vaginally, or topically.

In a specific embodiment of the present invention the pharmaceutical composition may be in the form of capsules or in the form of microcapsules, forming a powder or in the form of sachets of powder. The capsule can be flavored. This implementation also includes the capsule and the capsule and encapsulated composition of fatty acids corresponding to the present invention, scented. Flavored capsule is more attractive to the consumer. For the above-mentioned therapeutic uses the dosage will, of course, vary depending on the used compound, the route of administration, the desired treatment and the extent of violations of the organism.

The pharmaceutical composition may be formulated in such a way that the daily dosage will be from 10 mg to 10, Preferably the pharmaceutical composition is made up in such a way that the daily dosage of the above-mentioned composition ranges from 5 mg to 5, More preferably the pharmaceutical composition is made up in such a way that the daily dosage of the above-mentioned composition ranges from 100 mg to 1 g Under the daily dosage is the dosage within 24 hours. The dosage will, of course, vary depending on the used compound, the route of administration, telemag the treatment and extent of violations of the organism. The doctor usually determines the dosage which will be most suited to this subject. Dose and frequency of use for each individual patient can vary and will depend on many factors, including the activity of the used compounds, the metabolic stability and length of action of the compound, the age, body weight, General state of health, sex, diet, mode and time of administration, rate of excretion, combination of drugs, the severity of the pathological condition and individual therapy. Substance and/or pharmaceutical composition of the present invention can be introduced into the body from 1 to 10 times a day, for example 1 or 2 times a day. Oral and parenteral administration, the daily dose of a substance can be injected once or may be divided into multiple doses.

A further aspect of the present invention relates to the composition of fatty acids, including the compounds of formula (I). Composition of fatty acids, including the compounds of formula (I), increases the natural biological effects of DHA, which are the result of regulation of gene expression, and derivatives corresponding to the present invention, will be accumulated in the pool of free fatty acids.

Composition of fatty acids may contain from 60 to 100% by weight of compounds of formula (I), all percentages by weight osnovy who are on the total weight of the composition of fatty acids. In a preferred embodiment of the present invention at least 80% by weight of the composition of fatty acids are the compounds of formula (I). More preferably the compounds of formula (I) comprise at least 90% by weight of the composition of fatty acids. More preferably the compounds of formula (I) account for more than 95% by weight of the composition of fatty acids. Composition of fatty acids may include at least one of the fatty acids: (a fully-Z)-5,8,11,14,17-eicosapentaenoic acid (EPA), (a fully-Z)-4,7,10,13,16,19-docosahexaenoic acid (DHA),-Z)-6,9,12,15,18-hanakotoba acid (NDA) and (fully-Z)-7,10,13,16,19-docosapentaenoic acid (DPAn-3), (a fully-Z)-8,11,14,17-eicosatetraenoic acid (Stage-3), or combinations thereof. In addition, the composition of fatty acids may include (fully-Z)-4,7,10,13,16-docosapentaenoic acid (DPAn-6) and/or (fully-Z)-5,8,11,14-eicosatetraenoic acid (ARA) or their derivatives. Composition of fatty acids can also include these fatty acids or their combination in the form of derivatives. Derivatives substituted as derived DHA formula (I)as defined above.

Composition of fatty acids corresponding to the present invention, may include (a fully-Z omega-3)-6,9,12,15,18 - generateinmemory acid (NDA) or their derivatives in the amount of at least 1% by weight or share of the ve from 1 to 4% by weight.

In addition, the composition of fatty acids corresponding to the present invention, may include omega-3 fatty acids, not EPA and not DHA, which are 20, 21, or 22 carbon atoms, or their derivatives in the amount of at least 1.5% by weight in total of at least 3% by weight.

In specific embodiments, the implementation of the present invention, the composition of fatty acids is a pharmaceutical composition, a food composition or dietary composition. Composition of fatty acids can also include an effective amount of pharmaceutically acceptable antioxidant. Preferably the antioxidant is tocopherol or a mixture of Tocopherols. In a preferred embodiment, the composition of fatty acids, also contains tocopherol or a mixture of Tocopherols in an amount up to 4 mg per 1 g of the total weight of the composition of fatty acids. Preferably the composition of fatty acids include the amount of Tocopherols from 0.2 to 0.4 mg per 1 g of the composition.

Another aspect of the present invention relates to the composition of fatty acids, any pharmaceutically acceptable salt, MES, prodrug or complex, which include the compounds of formula (I)as defined above, for use as a medicine and/or therapy. The composition of fatty acids can be used to counter the surveillance and/or treatment of such pathological conditions, employ the compounds of formula (I)as described above.

When the composition of fatty acids is used as a medicine, it will be introduced into the organism a therapeutically or pharmaceutically active amount.

In a preferred embodiment, the composition of fatty acids in humans or animals is for oral administration.

The present invention relates also to the use of compounds of formula (I), or pharmaceutically acceptable salts of MES, prodrug or complex, as defined above, for the manufacture of a medicinal product for controlling body weight reduction and/or prevention of gaining weight; for the manufacture of a medicinal product for the treatment and/or prevention of obesity or overweight; for the manufacture of a medicinal product for the prevention and/or treatment of diabetes in humans or animals; for the manufacture of a medicinal product for the treatment and/or prevention of diseases associated with amyloidosis; for the manufacture of a medicinal product for the treatment or prevention multiple risk factors for cardiovascular diseases such as high blood pressure, hypertriglyceridemia and high coagulation activity of factor VII; for the manufacture of a medicinal product for the treatment of tuberculosis or HIV; Dneprospetsstal drugs for prevention of cerebral circulation, cerebral or transient ischaemic attacks related to atherosclerosis of some of the arteries; for the manufacture of a medicine for lowering triglycerides in the blood of mammals and/or increase the level of high density lipoprotein in patients; or for the manufacture of a medicinal product for the treatment and/or prevention of metabolic syndrome. All of these options for implementation include the use of a composition of fatty acids, as defined above, including the components of the formula (I) for the production of medicines, as mentioned above. The present invention also relates to the use of alpha-hydroxy-DHA for the production of medicines, as indicated above.

The present invention also relates to a method of controlling body weight reduction and/or prevention of gaining weight, where the composition of fatty acids comprising at least the compound of formula (I)as defined above, is administered to a human or animal.

In addition, the present invention relates to a method of treatment and/or prevention of obesity or overweight, where the composition of fatty acids comprising at least the compound of formula (I)as defined above, is administered to a human or animal.

In a preferred embodiment of the present invention present from Britanie relates to a method for prevention and/or treatment of diabetes, where the composition of fatty acids comprising at least the compound of formula (I)as defined above, is administered to a human or animal. Preferably diabetes mellitus is diabetes of the 2nd type.

Other aspects of the present invention relate to:

the method of treatment and/or prevention of diseases associated with amyloidosis;

the method of treatment or prophylaxis of multiple risk factors for cardiovascular disease;

the way to prevent violations of cerebral circulation, cerebral or transient ischaemic attacks related to atherosclerosis of certain arteries;

where the composition of fatty acids comprising at least the compound of formula (I)as defined above, is administered to a human or animal.

Derivatives of fatty acids of the formula (I) can be obtained most effectively on the basis of DHA. If the starting material is not pure DHA (i.e. not 100% DHA), the final composition of fatty acids will contain a mixture of derivatives of DHA, as defined above, and a number of other fatty acids, not DHA, where these fatty acids are substituted in the same way as new analogues of fatty acids of the formula (I). Such variants of implementation is also included in the present invention.

In another embodiment of the present invention the compounds of formula (I) obtained on the basis of (full the STU-Z)-4,7,10,13,16,19-docosahexaenoic acid (DHA), where specified DHA derived from a source of vegetable origin, source of microbial and/or animal origin or their combinations. Preferably specified DHA derived from fish oil.

Fatty acid composition can also be obtained from a source of vegetable, microbial or animal origin or their combination. Thus, the present invention also includes the composition of fatty acids prepared from microbial fat.

The present invention relates to a method for the preparation of new analogues of fatty acids of the formula (I)as defined above.

DHA derived from biological sources: fat sea fish, microbial fat, vegetable fat. All possible raw materials are mixtures of fatty acids in triglyceride form, where DHA is only a fraction of fatty acids. Typically, the concentration of DHA is 40% in microbial fat and 10-25% in the fat of marine fish. Contains DHA vegetable fats are in development, and in the future is expected to produce fats with a high concentration of DHA.

The first step of the process will always be the conversion of triglycerides into free fatty acids or monetary. Preferred esters are complex ethyl or methyl ester. Thus, fatty acids are bound in the triglyceride, are separated from one another, and OTDELENIE is made possible. There are several ways of separating DHA from other fatty acids, the most frequently used method of molecular distillation, in which the fatty acids are separated by evaporation, and the method of separation by precipitation with urea, in which the fatty acids are separated according to the degree of unsaturation. Separation of fatty acid is also produced by the following methods: method using silver nitrate, in which the fatty acids are separated according to the degree of unsaturation, the esterification reaction, catalyzed by lipases, in combination with molecular distillation and countercurrent extraction with carbon dioxide in supercritical state.

The most important task associated with obtaining the pure DHA. is a branch of this fatty acid from other highly unsaturated fatty acids containing from 20 to 22 carbon atoms, present in all sources. These fatty acids have properties that are so similar to DHA that none of the methods listed above, does not provide a sufficient degree of separation. For some microbial fats containing large amounts of DHA and very low levels of highly unsaturated fatty acids containing from 20 to 22 carbon atoms, the application of the method of molecular distillation alone or in combination with other methods can provide more than 90% purity.

Fats, with whom containing a series of large amount of DHA, also contain significant amounts of highly unsaturated fatty acids containing from 20 to 22 carbon atoms, such as EPA (20:5n-3), n-3DPA (22:5n-3), HPA (21:5n-3) and others. The only existing way of separating DHA from such fatty acids is high performance liquid chromatography as a stationary phase using silica gel or silica gel impregnated with silver nitrate as the mobile phase used is selected organic solvents or carbon dioxide in a supercritical state. Using this method, you receive DHA with more than 97% purity. However, it should be noted that the processing cost becomes high with increasing concentration, for example, a production cost 97% DHA more than five times the technological cost 90% DHA.

DHA, having a purity of 90, 95 or 97%, contains very small amounts of other fatty acids. For example, DHA, having a purity of 97%, contains n-3DPA (22:5n-3), but also long-chain fatty acids such as EPA (20:5n-3), HPA (21:5n-3) and others. However, other fatty acids will react in the same way as DHA and will be received alpha-substituted derivatives.

Using organic synthesis possible method of purification, so as DHA and n-6DPA (and 22:5n-6, which is generally present in very low concentrations) are the only known fat sour the AMI, on the basis of which you can obtain gamma-lactones by cyclization with the first double bond. Can be applied to the following path: lactonization, then cleaning and hydrolysis, but this method is more expensive than HPLC (high performance liquid chromatography).

In one embodiment, the compounds of formula (I), where R1(or R2is hydrogen obtained by using the following methods (Scheme 1). Accordingly adapted, these methods can be used to obtain compounds represented by the General formula (I), where R1and R2are, for example, With1-C7alkyl group, benzyl, halogen, benzyl, alkenyl or quinil.

Compounds represented by the General formula (I), where R1is hydrogen and R2stands With1-C7alkyl group, benzyl, halogen, benzyl, alkenyl, quinil obtained by reaction of DHA ester with a strong dinucleophiles base, i.e. with substances such as lithium Diisopropylamine or geometrization potassium/sodium in a solvent such as tetrahydrofuran, diethylether at temperatures from -60 to -78°C, to obtain enolate ether (way 1/1 st process).

(Scheme 1)

The ester enolate is reacted with an electrophilic reagent, such alkylhalides, examples of which I have are ethyl iodide, benzylchloride, allalou, examples of which are methyl benzoyl, carboxyl anhydride, an example of which is acetic anhydride, or an electrophilic reagent of halogenization, an example of which is N-torbenson sulfonamid (NFSI), etc. to obtain the monosubstituted derivative (way 2/2 nd process). Ester then hydrolyzed in a solvent, such as ethanol or methanol to the carboxylic acid derivative by adding a base such as lithium hydroxide/sodium in water at temperatures from 15 to 40°C.

Condensation of Clausena ethyl ester of DHA occurs during the processing of the ethyl ester of DHA strong base. This product condensation may possess interesting biological activity. Thus, in one embodiment, the present invention describes the condensation of (intermediate) products mentioned above, and the use of this product for the treatment and/or prevention of diseases of the present invention.

In another embodiment, compounds represented by the General formula (I), are synthesized using the following methods (Scheme 2).

(Scheme 2)

Compounds represented by the General formula (I), where R1is hydrogen and R2denotes hydroxy, alkoxygroup, acyloxy obtained p is the reaction of DHA ester with a strong dinucleophiles base, i.e. with substances such as lithium Diisopropylamine or geometrization potassium/sodium in a solvent such as tetrahydrofuran, diethylether at temperatures from -60 to -78°C, to obtain enolate ether (4 way/process 4). This ester enolate reacts with this oxygen source, as dimethyldioxirane, 2-(phenylsulfonyl)-3-phenyloxazolidine, molecular oxygen with various additives, as trimethylphosphite or different catalysts like Ni(II) complex ester of alpha-hydroxy-DHA ester (method 5/process 5). The reaction of the secondary alcohol with a base such as sodium hydride in a solvent like THF or DMF, gives an alkoxide, which reacts with various electrophilic reagents such as alkyl iodide, for example; methyl iodide, ethyl iodide, benzyl bromide or Allgood, for example; acetylchloride, benzoyl bromide (method 6/process 6). Ester then hydrolyzed in a solvent, such as ethanol or methanol to the carboxylic acid derivative by adding a base such as lithium hydroxide/sodium in water at temperatures from 15 to 40°C (method 7/process 7).

Ether hydroxy-DHA is a convenient intermediate connection for the introduction of other functional groups in α-position to the present invention. Pyroxyline group can be activated by conversion to the halide or tosyl the t to react with various nucleophilic compounds, such as ammonia, amines, thiols, etc. For the conversion of hydroxyl groups into other functional groups can also be applied reaction Mitsunobu (Mitsunobu, Oh, Synthesis, 1981, 1).

Compounds represented by the General formula (I)as defined above, can be synthesized by various combinations of the described methods. The present invention includes the above-mentioned methods.

The present invention also relates to a method for producing a pharmaceutical composition of the present invention, which includes at least a compound of formula (I) or a pharmaceutically acceptable salt, MES, complex or prodrug, as defined above, with a pharmaceutically acceptable auxiliary substance, a solvent or carrier.

Clean entiminae compounds can be obtained by splitting of racemic compounds of formula (I)as defined above. Cleavage of compounds of formula (I) can be carried out using known procedures, for example, by reacting the compounds of formula (I) is the mixture of diastereoisomers, which can be separated by chromatography. After that, the two enantiomers of the compound (I) can be obtained from the divided diastereoisomers, for example, by hydrolysis.

You can also apply stoichiometric chiral compounds OS is enforced asymmetric introduction of substituents in the α-position DHA. The most effective method was the use of chiral oxazoline-2-it. Enolate obtained from chiral N-allocationof, can be blocked by various electrophiles in high stereoregularity manner (Ager, Prakash, Schaad 1996).

Examples

The present invention will be described in more detail using the following examples, which should not be seen as only possible in the present invention. In the examples of the structures were verified by mass spectrometry (MS). It should be noted that derivatives of fatty acids can also be obtained from a source material containing low and medium amounts of DHA (i.e. approximately 40-60% by weight DHA).

The synthesis protocols

Obtaining the ethyl ester of α-methyl-D (PRB-1)

Utility (228 ml of 0.37 mol, 1.6 M in hexane) was added in drops to a stirred solution of Diisopropylamine (59,5 ml, 0.42 mol) in dry THF (800 ml) in an atmosphere of N2at 0°C. the resulting solution was stirred at 0°C for 30 minutes, cooled to -78°C. and then was stirred for an additional 30 minutes before adding dropwise DHA ethyl ester (100 g, 0.28 mol) in dry THF (500 ml) for 2 hours. The dark green solution was stirred at -78°C for 30 minutes before adding Me (28 ml, 0.45 mol). The solution was left for 1.5 hours until they reach the temperature of -20°C, then infused in water is (1.5 l) and extracted with heptane (2×800 ml). The combined organic phases were washed 1 M Hcl (1 l), dried (Na2SO4), filtered and evaporated in vacuum. The product was purified by flash chromatography on silica gel, elution was carried out using heptane/tO (99:1), and was obtained 50 g (48%) of the named compound as a pale yellow oil;

1H-NMR (200 MHz, CDCl3) δ of 1.02 (t, J 7.5 Hz, 3H), 1,20 (d, J 6.8 Hz, 3H), of 1.29 (t, J 7,1 Hz, 3H), of 2.0 to 2.6 (m, 5H), of 2.8-3.0 (m, 10H), 4,17 (t, J 7,1 Hz, 2H), 5,3-5,5 (m, 12H);

MC (electrospray ionization); 393 [M+Na].

Obtaining the ethyl ester of α-ethyl-DHA (PRB-2)

Utility (440 ml, 0.67 mol, 1.6 M in hexane) was added in drops to a stirred solution of Diisopropylamine (111 ml, 0.78 mol) in dry THF (750 ml) in an atmosphere of N2at 0°C. the resulting solution was stirred at -78°C for 45 minutes before adding dropwise DHA ethyl ester (200 g, of 0.56 mol) in dry THF (1.6 l). The addition of ester was completed after 4 hours. The dark green solution was stirred at -78°C for 30 minutes before adding the EtI (65 ml, 0.81 mol). The solution was left to reach 40°C before adding additional quantities EtI (5 ml, 0.06 mol), and then the solution was left up to the age of 15°C (within 3 hours from -78°C), then the mixture was infused into water and extracted with hexane (2x). The combined organic phases were washed 1 M Hcl, water, dried (Na2SO4), Hotfile is delicate and evaporated in vacuum. The product was purified by flash chromatography on silica gel, elution was carried out using heptane/tO (99:1 then 50:1), and was obtained at 52.2 g (20%) of the named compound as a yellow oil.

1H-NMR (200 MHz; CDCl3) δ of 0.8-1.0 (m, 6H), 1,2-1,4 (m, 4H), of 1.5-1.7 (m, 2H), 2,12 (m, 2H), 2,3-2,5 (m, 2H), 2.8 to 3.0 (m, 10H), 4,18 (t, J=7,1 Hz, 2H), 5,3-5,6 (m, 12H);

MC (electrospray ionization); 407 [M+Na].

Obtaining the ethyl ester of α-ethoxy-D (PRB-3)

To a suspension of 60% NaH (84,1 mg, 2.1 mmol) in THF, 5 ml) at -78°C in an atmosphere of N2was added in drops a solution of the ethyl ester of α-hydroxy-DHA (PRB-12) (372 mg, 1.00 mmol) in THF, 5 ml, the mixture was stirred at -78°C for 20 minutes before adding drops of ethyl iodide (of 0.24 ml, a 3.01 mmol). The reaction mixture was gradually brought to room temperature over night. Was added a saturated aqueous solution of NH4Cl, 15 ml, and the mixture was extracted with diethyl ether, 25 ml × 2), organic phase was washed with brine, 25 ml, dried (Na2SO4), filtered and evaporated in vacuo, then was held flash chromatography on silica gel, elution was carried out using heptane/tO (95:5), and was obtained 68 mg (17%) of product as a yellow liquid.

1H NMR (200 MHz, CDCl3) δ were 0.94 (t, J=7.5 Hz, 3H), 1,16-of 1.29 (m, 6H), was 2.05 (quintet, J=7.2 Hz, 2H), 2,50 (m, 2H), was 2.76-2,84 (m, 10H), 3.33 and-of 3.48 (m, 1), 3,53-3,71 (m, 1H), 3,83 (DD, J=6,8 Hz, J=6.2 Hz, 1H), 4,18 (q, J=7,1 Hz, 2H), 5,31-of 5.45 (m, 12H)

13With NMR (50 MHz, Dl3) δ 14,2, 15,1, 20,5, 25,5, 25,6, 25,7, 31,0, 60,8, 66,0, 78,7, 124,1, 127,0, 127,8, 127,9, 128,0 (2 signal), 128,2 (2 signal), 128,5, 130,7, 132,0, 172,5 (3 hidden signal)

MC (electrospray ionization); 423 [M+Na]+

Obtaining the ethyl ester of α-fluorescent-DHA (PRB-4)

To LDA (2.1 ml, 4.2 mol 2 M in THF/heptane/ethylbenzene) in dry THF (10 ml) in an atmosphere of N2at -78°C was added in drops within 15 minutes DHA ethyl ester (1 g, 2.8 mmol) in dry THF (30 ml). Then was added NFSi (1.06 g, 3.4 mmol). The solution was left until they reach room temperature and then stirred for 70 hours. The mixture was infused into water and extracted with hexane (2x). The combined organic phases were washed 1 M HCl, water, dried (Na2SO4), filtered and evaporated in vacuum; MS (electrospray ionization); 397 [M+Na].

Obtaining the ethyl ester of α,α-dimethyl DHA (PRB-5)

Utility (100 ml of 0.17 mol, 1.6 M in hexane) was added in drops to a stirred solution of Diisopropylamine (28 ml, 0.20 mol) in dry THF (100 ml) in an atmosphere of N2at 0°C. the resulting solution was stirred at 0°C for 30 minutes, cooled to -78°C and drops was added a solution of DHA ethyl ester (50 g, 0.14 mol) in dry THF (200 ml). The obtained dark green solution is stirred at -78°C for 30 minutes before adding Me (17 ml, 0.28 and the ol). The solution was left until they reach a temperature of -10°C, and then infused into water and extracted with heptane (2). The combined organic phases were washed 1 M Hcl (1 l), dried (Na2SO4), filtered and evaporated in vacuum.

The procedure was repeated, but the crude product ethyl ester of α-methyl-DHA was used instead of the ethyl ester of DHA. The product was purified by flash chromatography on silica gel, heptane/tO (99:1, then 98:2), and was obtained 31.6 g (59%) of the named compound as a pale yellow oil;

1H-NMR (200 MHz, CDCl3) δ is 1.01 (t, J 7.5 Hz, 3H), of 1.21 (s, 6H), of 1.28 (t, J 7,1 Hz, 3H), of 2.08 (m, 2H), 2,34 (d, J 6.8 Hz, 2H), 2.8 to 3.0 (m, 10H), is 4.15 (q, J 7.5 Hz, 2H), 5,3-5,6 (m, 12H);

13C-NMR (50 MHz, CDCl3) δ 14,7, 21,0, 25,3, 26,0, 26,1, 38,3, 42,8, 60,7, 125,8, 127,4, 128,3, 128,5, 128,6, 128,7, 129,0, 130,7, 132,4, 177,9;

MS (electrospray ionization); 385 [M+H].

Obtaining the ethyl ester of α-thiomethyl-D (PRB-6)

Ethyl ester of α-iodide-DHA (0.5 g, 1.04 mmol) was dissolved in 20 ml of THF in an atmosphere of N2at 0°C. was added MeSNa (80 mg, to 1.14 mmol) and the mixture was stirred for a few minutes before it was diluted with heptane. The organic phase was washed with water (2x), dried (Na2SO4) and evaporated in vacuo. The desired product was purified by flash chromatography, elution was carried out using heptane/tO (30:1), was obtained ethyl ester of α-thiomethyl-DHA in the form of pale yellow is oil. Ethyl ester of α-thiomethyl-DHA was dissolved in 10 ml of EtOH and 10 ml F. To the solution was added LiOH (0.39 g, 9.2 mmol), dissolved in 5 ml of water. The reaction mixture was stirred over night at room temperature, then the mixture was diluted with water and heptane.

The organic phase was extracted with 1 M LiOH (2x) and the combined aqueous phases were acidified with 5 M Hcl and extracted with diethyl ether (2x). The combined organic phases were washed with brine, water, dried (Na2SO4) and evaporated in vacuo, the resulting 183 mg (47%) of the titled compound as a pale yellow oil;

1H-NMR (200 MHz, CDCl3) δ and 0.98 (t, J 6.6 Hz, 3H), 1,95-to 2.65 (m, 7H), 2,72 was 3.05 (m, 10H), 3,12-of 3.43 (m, 1H), 5,20-5,70 (m, 12H), 10,65 (br s, 1H);

13H-NMR (50 MHz, Dl3) δ 14,7, 21,0, 25,9, 26,0, 26,2, 28,8, 125,4, 127,4, 128,1, 128,3, 128,4, 128,7, 128,9, 129,0, 131,6, 132,4, 177,0.

Obtaining the ethyl ester of α-thioethyl-DHA (PRB-7)

Ethyl ester of α-iodide-DHA (11 g, 23 mmol) was dissolved in 100 ml of THF in an atmosphere of N2at 0°C.

Was added EtSNa (2.1 g, 25 mmol) and the mixture was stirred for 1 hour at 0°C. the Reaction was stopped using 1 M Hcl and dilution was carried out with heptane. The organic phase was washed with water (2x), dried (Na2SO4) and evaporated in vacuo. The desired product was purified by flash chromatography, heptane/tO (30:1), and was obtained of 7.3 g (76%) named what about the connection in the form of a pale yellow oil;

1H-NMR (200 MHz, CDCl3) δ of 1.1-1.3 (m, 9H), was 2.05 (m, 2H), 2,3-2,7 (m, 4H), 2,7-2,9 (m, 10H), of 3.25 (m, 1H), 4,17 (q, J 7,1 Hz, 2H), 5,3-5,5 (m, 12H);

MS (electrospray ionization): 439 [M+Na].

Obtaining the ethyl ester of α,α-diethyl-D (PRB-8):

Utility (38,6 ml of 0.62 mol, 1.6 M in hexane) was added in drops to a stirred solution of Diisopropylamine (9.1 ml of 0.65 mol) in dry THF (200 ml) in an atmosphere of N2at 0°C. the resulting solution was stirred at 0°C for 30 minutes, cooled to -78°C and drops was added a solution of ethyl ester of DHA (20,0 g, of 0.56 mol) in dry THF (100 ml). The obtained dark green solution is stirred at -78°C for 30 minutes before adding the EtI (6.8 ml, 0.84 mol). The solution was left until they reach a temperature of -10°C, and then infused into water and extracted with hexane (2x). The combined organic phases were washed 1 M Hcl, dried (Na2SO4), filtered and evaporated in vacuum.

The procedure was repeated, but the crude product ethyl ester of α-ethyl-DHA was used instead of the ethyl ester of DHA. The reaction mixture after addition of EtI was left to reach room temperature and stirred over night. The product was purified by flash chromatography on silica gel, elution was carried out using heptane/tO (99:1, then 98:2), and was obtained 10.0 g (43%) of the named compound as a pale yellow oil;

H-NMR (200 MHz, CDCl3) δ of 0.83 (t, J 7.4 Hz, 6H), were 0.94 (t, J 5.8 Hz, 3H), of 1.28 (t, J 7,1 Hz, 3H), and 1.63 (q, 77,4 Hz, 4H), 2,10 (m, 2H), 2,34 (d, J 6.9 Hz, 2H), 2.8 to 3.0 (m, 10H), is 4.15 (q, J 7.5 Hz, 2H), 5,3-5,6 (m, 12H);

13C-NMR (50 MHz, CDCl3) δ 8,9, 14,7, 21,0, 23,1, 25,9, 26,0, 26,2, 27,4, 31.2, 50,1, 60,6, 125,5, 127,4, 128,3, 128,6, 128,9, 130,5, 132,4, 177,1;

MS (electrospray ionization); 413.3 [M+H], 435.3 [M+Na].

Obtaining the ethyl ester of α-benzyl-D (PRB-9)

To a stirred solution of Diisopropylamine (of 0.91 ml, 6,46 mmol) in dry THF (20 ml) under inert atmosphere at 0°C was added drops of n-BuLi (of 1.6 M in hexane, 3,86 ml, 6,18 mmol). The mixture was stirred at 0°C for 30 minutes, brought to -78°C and then stirred at this temperature for 5 minutes. Ethyl ester of DHA (2.0 g, 5,62 mmol) in dry THF (10 ml) was added dropwise and the mixture is stirred at -78°C for 20 minutes, then was added benzyl bromide (0,80 ml, 6,74 mmol). The resulting solution was kept for 3 h while the temperature was 0°C, then subjected distribution between water (100 ml) and heptane (100 ml). The aqueous layer was extracted with heptane and the combined organic layers were washed with 1 M Hcl and dried (Na2SO4). Was concentration under reduced pressure and purification via flash chromatography (Heptane: tO 99:1), the result of 1.05 g (42%) of the named compound as a colourless oil;

1H-NMR (200 MG IS, CDCl3): δ 0,99 (t, 3H), of 1.18 (t, 3H), 2,08-of 2.16 (m, 2H), 2,35-to 2.42 (m, 2H), 2,74 are 2.98 (m, 13H), 4.09 to (q, 4H), 5,38-5,50 (m, 10H), 7,19 and 7.36 (m, 5H);

13C-NMR (50 MHz, CDCl3): δ 14,61, 14,71, 20,99, 25,98, 26,07, 30,07, 38,32, 48,02, 60,88, 126,75, 126,83, 127,46, 128,31, 128,45, 128,53, 128,58, 128,86, 128,77, 129,01, 129,35, 130,55, 132,46, 138,89, 175,39.

MS (electrospray ionization): 447.3 [M+H], 469.3 [M+Na].

Obtaining the ethyl ester of α-econsultancy-D (PRB-10)

To a solution of ethyl ester of α-thioethyl-DHA (0.5 g, 1.3 mmol) in 15 ml l3in an atmosphere of N2at -20°C was added a solution of MSRV (0,22 g, 1.3 mmol) in 10 ml l3. The reaction mixture was stirred for 2 hours at this temperature, then the mixture was filtered and washed with saturated aqueous Panso3. The aqueous phase was extracted twice with HCl3and the combined organic phase was washed with water and with brine, dried over Na2SO4filtered and concentrated. The product was purified by flash chromatography using hexane: tO 8:2, was obtained 0.35 g (70%) of these connections.

1H NMR (200 MHz, CDCl3): δ 0,99 (t, 3H), 1,27-of 1.45 (m, 6H), is 2.09 (m, 2H), 2,79-to 2.94 (m, 14H), 3,55 (m, 1H), 4,25 (q, 2H), lower than the 5.37-5,59 (m, 12H).

13C-NMR (50 MHz, CDCl3): δ 7,97, 14,58, 14,68, 20,95, 23,68, 25,17, 25.93, 26,04, 44,20, 45,15, 62,30, 64,08, 123,91, 124,47, 127,41, 127,86, 128,26, 128,40, 128,44, 128,72, 128,72, 128,96, 129,12, 132,42, 132,47, 174,55.

MS (electrospray ionization): 455.3 [M+Na].

Getting ethyl the new ester of α-thiophenyl-D (PRB-11)

To a solution of ethyl ester of α-iodide-DHA (PRB-15) (3,40 g, 7.05 mmol) in acetone (20 ml) was added in drops a solution of sodium persulfide (1,039 g, 7,86 mmol) in acetone (110 ml). The resulting mixture was stirred at room temperature for half an hour, the mixture was evaporated in vacuo, then was held flash chromatography on silica gel, elution was carried out using heptane/tO 200:1-95:5, and the result of 2.35 g (72%) of product as a yellow liquid.

1H NMR (200 MHz, CDCl3): δ 0,97 (t, J=7.5 Hz, 3H), of 1.18 (t, J=7,1 Hz, 3H), of 2.09 (quintet, J=7,l Hz, 2H), 2,54-of 2.66 (m, 2H), 2,83-of 2.86 (m, 10 H)to 3.67 (dd, J=6,8 Hz, J=8,3 Hz, 1H), 4,12 (quintet, J=7,1 Hz, 2H), 5,24-5,49 (m, N), 7,28-7,33 (m, 3H), 7,46 is 7.50 (m, 2H)

13With NMR (50 MHz, Dl3): δ 14,0, 14,2, 20,5, 25,5, 25,6. 25,7, 29,4, 50,6, 61,1, 125,1, 127,0, 127,7, 127,9, 128,0, 128,3, 128,42, 128,45, 128,9, 131,2, 132,0, 133,0, 133,2, 174,1 (5 hidden signals)

MS (electrospray ionization); 465 [M+H]+, 487 [M+Na]+

Mass spectroscopy high resolution (HRMS) (EI (ionization by electron impact)) calculated for C30H40O2S: 464.2749 found: 464.2741

Obtaining the ethyl ester of α-hydroxy-D (PRB-12)

To a solution of Diisopropylamine (19,76 ml, 140 mmol) in dry THF, 40 ml, in an atmosphere of N2at -78°C was added drops of 1.6 M BuLi in hexane (87,5 ml, 140 mmol). The resulting mixture was stirred at -78°C for 15 minutes before those who, as the drops was added a solution of ethyl ester of DHA (24,99 g, 70,1 mmol) in THF (80 ml). The reaction mixture is dark green was stirred for 1 hour at -78°C. before it was added drops triethylphosphite (12,2 ml, 70,1 mmol) and then O2was passed through the reaction mixture overnight, the reaction mixture was maintained at -78°C for 5 hours and then slowly warmed to room temperature. Was added a saturated aqueous solution of NaHCO3(100 ml)and the mixture was extracted with diethyl ether (200 ml × 2. The organic phase was dried (Na2SO4), filtered and evaporated in vacuo, then was held flash chromatography on silica gel, elution was carried out using heptane/tO 99:1-95:5, and the resulting 4.52 g (17%) of product as a yellow liquid.

1H NMR (200 MHz, CDCl3): δ to 0.92 (t, J=7.5 Hz, 3H), 1,24 (t, J=7,1 Hz, 3H), 2,02 (quintet, J=7,1 Hz, 2H), 2,44-of 2.54 (m, 2H), 2,74-2,87 (m, 10 H), 4,13-4,24 (m, 3H), 5.25 to 5,94 (m, 12H)

13C-NMR (50 MHz, CDCl3): δ 14,0, 14,1, 20,4, 25,4, 25,5, 25,6, 32,0, 61,5, 69,9, 123,3, 126,9, 127,7, 127,9, 128,08, 128,1, 128,2, 128,4, 131,3, 131,8, 174,4 (4 hidden signal)

MS (electrospray ionization); 395 [MH-Na]+

HRMS (ES) calculated for C24H36O3Na: 395.2556 found: 395.2543

Getting amide α-methyl-D (PRB-13)

To a solution of α-methyl-DHA (PRB-1 FA) (3.13 g, 9.1 mmol) and oxalicacid (8,0 is l, the 94.5 mmol) in toluene (90 ml) was added DMF (0.1 ml) and the resulting mixture was stirred at room temperature in an atmosphere of N2within 15 ½ hours. The mixture was then evaporated in vacuo and the residue was dissolved in THF, 100 ml, the solution is cooled to 0°C, then drops was added aqueous NH3(20 ml). Tub with ice was removed and the mixture was stirred at room temperature for 4 hours, then was added 50 ml of water, and the aqueous phase was extracted with diethyl ether, 2×100 ml Organic phase was washed with saturated aqueous NH4Cl, 50 ml, dried (Na2SO4), filtered and evaporated in vacuo, then was held flash chromatography on silica gel, elution was carried out using a CH2CL2/2M NH3in the Meon 97.5:2.5, and the result of 2.51 g (80%) of product as a yellow liquid.

1H NMR (200 MHz, CDCl3) δ of 0.91 (t, J=7.5 Hz, 3H), 1,10 (d, J=9.8 Hz, 3H), 1,94-2,11 (m, 3H), 2,19 to 2.35 (m, 2H), was 2.76-2,77 (m, 10 H), 5,18-of 5.45 (m, 12 H), 6,03 (s, 1H), 6,72 (s, 1H)

13C NMR (50 MHz, CDCl3) δ 14,6, 17,6, 20,8, 25,8, 25,9, 32,0, 41,0, 127,3, 128,1, 128,4, 128,6, 128,8, 130,1, 132,2, 179,6 (8 hidden signals)

MS (electrospray ionization); 342 [M+H]+, 364 [M+Na]+

HRMS (EI) calculated for C23H35NO: 341.2719 found: 341.2707

Obtaining the ethyl ester of α-methoxy-DHA (PRB-14)

To a suspension of 60% NaH (61,1 mg, 1.53 mmol) in THF (5 ml) is tmosphere N 2at -78°C was added by drops a solution of the ethyl ester of α-hydroxy-DHA (PRB-12) (373 mg, 1.00 mmol) in THF (5 ml), the mixture was stirred at -78°C for 20 minutes before the drops was added methyl iodide (0,13 ml of 2.09 mmol). The reaction mixture was gradually brought to room temperature for 5 hours. Was added a saturated aqueous solution of NH4Cl, 15 ml, and the mixture was extracted with diethyl ether (25 ml × 2), the organic phase was washed with brine (25 ml), dried (Na2SO4), filtered and evaporated in vacuo, then was held flash chromatography on silica gel, elution was carried out using heptane/tO (99:1-4:1), and was obtained 136 mg (35%) of product as a yellow liquid.

1H NMR (200 MHz, CDCl3) δ to 0.92 (t, J=7.5 Hz, 3H), 1,24 (t, J=7,1 Hz, 3H), 2,03 (quintet, J=7,3 Hz, 2H), 2,48 (t, J=5.7 Hz, 2H), 2,73-2,82 (m, 10 H)to 3.34 (s, 3H), 3,74 (t, J=6.2 Hz, IH), 4,17 (q, J=7,1 Hz, 2 H), 5,24-5,43 (m, 12H)

13C NMR (50 MHz, CDCl3) δ 14,1, 20,4, 25,4, 25,5, 25,7, 30,6, 57,9, 60,9, 80,8, 123,7, 126,9, 127,71, 127,73, 127,92, 127,94, 128,07, 128,1, 128,2, 128,4, 130,7, 131,8, 171,9 (3 hidden signal)

MS (electrospray ionization); 409 [M+Na]+

HRMS (ES) calculated for C25H38O3Na: 409.2713 found: 409.2711

Obtaining the ethyl ester of α-iodide-DHA (PRB-15)

Diisopropylamine (20 ml, 140 mmol) was dissolved in 150 ml of THF in an atmosphere of N2at -20°C. BuLi (88 ml, 140 mmol, 1,6 M) was drops add the updates to the mixture before as the solution was cooled to -78°C. Ethyl ester of DHA (50 g, 140 mmol) in 250 ml drops of THF was added to the solution and the reaction mixture was stirred for 30 minutes at room temperature. The resulting mixture was added in drops to a solution of I2(42.8 g, 169 mmol) in 400 ml THF in an atmosphere of N2at -70°C. the Reaction was stopped using 1 M Hcl and dilution was carried out with heptane. The organic phase was washed with 10% Na2S2O3(2x), dried (Na2SO4), filtered and evaporated in vacuum. The desired product was purified using flash-chromatography, as eluent was used Heptane/tO (100:1), and was obtained 11.0 g (16%) of the named compound as a pale yellow oil;

MS (electrospray Ionization): 505 [M+Na].

Obtaining the ethyl ester of α-iodide-D (PRB-15)

To a solution of Diisopropylamine (42 ml, 298 mmol) in dry THF (150 ml) in an atmosphere of N2at -78°C drops was added 1.6 M BuLi in hexane (158 ml, 253 mmol). The resulting mixture was stirred at -78°C for 35 minutes, then was added by drops a solution of ethyl ester of DHA (75,05 g, 210 mmol) in THF (300 ml). The obtained dark green reaction mixture was stirred for 30 minutes at -78°C before adding drops of a solution of I2(91,06 g, 359 mmol) in THF (200 ml). The reaction mixture was stirred at -78°C for 2 minutes, then the reaction was stopped with water (200 ml), extraction was carried out with heptane (300 ml). The organic phase was washed with 1 M Hcl (150 ml) and water (200 ml), dried (Na2SO4), filtered and evaporated in vacuum. The crude product was purified by the method of flash-chromatography on silica gel, as eluent was used heptane/tO (100:1), and was obtained 26,14 g (26%) of product as a yellow liquid.

1H NMR (200 MHz, CDCl3) δ were 0.94 (t, J=7.5 Hz, 3H), 1,24 (t, J=7,1 Hz, 3H), 2,04 (quintet, J=7,1 Hz, 2H), 2,69-2,84 (m, 12 H)to 4.17 (q, J=7,l Hz, 2H), 4,22 (t, J=7.9 Hz, 1H), 5,24-5,49 (m, 12 H)

13With NMR (50 MHz, CDCl3) δ 13,7, 14,2, 25,5, 26,0 (2 signal), 25,8, 34,0, 61,7, 126,1, 127,0, 127,4, 127,8, 127,9, 128,0, 128,2, 128,5, 128,5, 131,6, 131,9, 170,9 (4 hidden signal)

MS (electrospray ionization); 505 [M+Na]+

Obtaining the ethyl ester of α-amino-D (PRB-17)

To a solution of ethyl ester of α-phthalimide-DHA (313,5 mg of 0.62 mmol) in ethanol (5 ml) was added hydrazine hydrate (46 μl, 0.95 mmol) and the resulting mixture was delegirovali in the atmosphere N2within 15 ½ hours, followed by evaporation in vacuo, purification was carried out using the method of the flash-chromatography on silica gel, elution was carried out using a CH2CL2:7M NH3in methanol (99:1-95:1), the result is 149 mg (64%) of product as a yellow liquid.

1N MR (200 MHz, CDl 3) δ of 0.91 (t, J=7.5 Hz, 3H), 1,22 (t, J=7,1 Hz, 3H), 1,72 (bs, 2H), 2,02 (quintet, J=7.2 Hz, 2H), 2,39 is 2.46 (m, 2H), 2,73-2,82 (m, 10 H), 3,47 (bs, 1H), 4,13 (q, 2H), 5,23-5.56mm (m, 12 H)

13With NMR (50 MHz, CDCl3) δ 14,1, 20,4, 25,4, 25,5, 25,6, 54,1, 60,8, 124,4, 126,9, 127,7 (2 signal), 127,9, 128,2, 128,3, 128,4, 131,4, 131,9, 189,3 (6 hidden signals)

MS (electrospray ionization); 372 [M+H]+

Obtaining the ethyl ester of (S)-(+)-α-ethyl DHA (PRB-20)

Synthesis of intermediate compound PRB-18:

DHA (3.00 g, and 18.3 mmol) was dissolved in dry CH2CL2(120 ml) at 0°C in an inert atmosphere and were added DMAP (2,45 g of 20.1 mmol) and DCC (3,96 g, 19.2 mmol). The mixture was stirred at 0°C for 20 minutes, then was added (4R,5S)-(+)-4-methyl-5-phenyl-2-oxazolidinone (3,24 g, and 18.3 mmol) and the resulting mixture was stirred at room temperature for 20 hours. The product was filtered and purified by the method of flash chromatography (heptane: ethyl acetate 6:1) gave yield 3.00 g (34%) of intermediate compounds PRB-18 as a colourless oil.

1H-NMR (200 MHz, CDCl3): δ 0,93-1,05 (t+d, 6H), 2,11 (m, 2H), of 2.51 (m, 2H), 2,80-3,00 (m, 10H), was 3.05 (m, 2H), 4,77 (m, 1H), 5,34-of 5.68 (m, 12H), 5,70 (d, 1H), 7,28-to 7.32 (m,2H), 131-1 Al (m, 3H).

Synthesis of intermediate compound PRB-19

Connection PRB-18 (1.80 g, 3,70 mmol) in dry THF (10 ml) was added in drops to a solution of LiHMDS (1M in THF, of 4.00 ml, 4.00 mmol) in dry THF (15 ml) at -78°C in an inert atmosphere. The mixture was stirred at-78°C for 30 minutes, was added EtI (0,89 ml, 11.1 mmol)and the mixture for one hour slowly brought up to 0°C. Then the mixture was stirred at 0°C for 18 hours and subjected distribution between saturated NH4C1 (50 ml) and diethyl ether (50 ml). The aqueous layer was extracted with diethyl ether (50 ml), the combined organic layer was washed with 0.1 M HCl (50 ml) and with brine (50 ml). Drying was carried out over Na2SO4the cleaning method flash chromatography (heptane: ethyl acetate 95:5) gave yield 0.52 g (27%) of intermediate compounds PRB-19 in the form of a colorless oil.

1H-NMR (200 MHz, CDCl3): δ 0.88 to a 1.01 (m, 9H), 1,64-of 1.78 (m, 2H), 2,08 (m. 2H), 2,31 (m, 1H), 2,48 (m, 1H), 2,87 (m, 10H), a 3.87 (m, 1H), and 4.75 (m, 1H), 5,32 (m, 12H), 5,63 (d,.77,1 Hz, 1H), 7,32 (m, 2H), 7,42 (m, 3H).

13C-NMR (50 MHz, CDCl3): δ 7,26, 11,75, 14,67, 14,98, 20,95, 25,57, 25,93, 26,04, 29,93, 44,59, 55,31, 79,10, 125,21, 126,01, 127,17, 127,42, 128,27, 128,50, 128,55, 128,67, 128,95, 129,09, 130,35, 132,42, 133,80, 153,18, 176,25.

MS (electrospray ionization): 538.2 [M+Na]+

Connection PRB-19 (0.25 g, 0,485 mmol) was dissolved in absolute ethanol (5 ml) and the temperature is brought to 0°C in an inert atmosphere. Was added NaOEt (1 M in ethanol, 0.54 ml, 0.54 mmol) and the mixture was stirred at 0°C for 30 minutes and subjected distribution between water and heptane. The aqueous layer was extracted with heptane and the combined organic layer was washed with 0.1 M HCl and dried. Cleanup of meth is remote flash chromatography gave the yield 0.025 g (13%) of the named compound PRB-20 in the form of a colorless oil.

1H-NMR (200 MHz, CDCl3) δ of 0.8-1.0 (m, 6H), 1,2-1,4 (m, 4H), of 1.5-1.7 (m, 2H), 2,12 (m, 2H), 2,3-2,5 (m, 2H), 2.8 to 3.0 (m, 10H), 4,18 (t, 2H), 5,3-5,6 (m, 12H).

MS (electrospray ionization); 407 [M+Na].

[α]D+1,7°(C=1.5, ethanol).

Obtaining the ethyl ester of (R)-(-)-α-ethyl-D (PRB-21):

Synthesis of intermediate compound PRB-21:

DHA (1,00 g of 3.05 mmol) was dissolved in dry CH2CL2(20 ml) at 0°C in an inert atmosphere and were added DMAP (0,41 g, 3.35 mmol) and DCC (0.66 g, 3,20 mmol). The mixture was stirred at 0°C for 20 minutes was added(4S,5R)-(-)-4-methyl-5-phenyl-2-oxazolidinone (0.54 g, of 3.05 mmol) and the mixture was stirred at room temperature for 20 hours. Filtering and purification by the method of flash chromatography (heptane: ethyl acetate 6:1) gave a yield of 1.08 g (73%) of intermediate compounds PRB-21 in the form of a colorless oil.

1H-NMR (200 MHz, CDCl3): δ 0,93-1,05 (t+d, 6H), 2,11 (m, 2H), of 2.51 (m, 2H), 2,80-3.00 (m, 10H), was 3.05 (m, 2H), 4,77 (m, 1H), 5,34-of 5.68 (m,12H), 5,70 (d, 1H), 7,28.to 7.32 (m, 2H), 7,37-7,47 (m, 3H).

Synthesis of intermediate compound PRB-22:

Connection PRB-21 (3.25 g, to 6.67 mmol) in dry THF (15 ml) was added in drops to a solution of LiHMDS (1M in THF, 7,34 ml, 7,34 mmol) in dry THF (35 ml) at -78°C in an inert atmosphere. The mixture was stirred at -78°C for 30 minutes, was added EtI (1.6 ml, 20.0 mmol) and the mixture for one hour slowly brought up to 0°C. Then the mixture was stirred at 0°C for 18 cha is s and subjected distribution between saturated NH 4Cl (50 ml) and diethyl ether (50 ml). The aqueous layer was extracted with diethyl ether (50 ml), the combined organic layer was washed with 0.1 M HCl (50 ml) and with brine (50 ml). Drying was carried out over Na2SO4the cleaning method flash chromatography (heptane: ethyl acetate 95:5) gave yield 1.50 g (44%) intermediate PRB-22 in the form of a colorless oil.

1H-NMR (200 MHz, CDCl3): δ 0.88 to a 1.01 (m, 9H), 1,64-of 1.78 (m, 2H), 2,08 (m, 2H), 2,31 (m, 1H), 2,48 (m, 1H), 2,87 (m, 10H), a 3.87 (m, 1H), and 4.75 (m, 1H), 5,32 (m, 12H), 5,63 (d, J7,l Hz, 1H), 7,32 (m, 2H), 7,42 (m, 3H).

13C-NMR (50 MHz, CDCl3): δ 7,26, 11,75, 14,67, 14,98, 20,95, 25,57, 25,93, 26,04, 29,93, 44,59, 55,31, 79,10, 125,21, 126,01, 127,17, 127,42, 128,27, 128,50, 128,55, 128,67, 128,95, 129,09, 130,35, 132,42, 133,80, 153,18,176,25.

MS (electrospray ionization): 538.2 [M+Na]

Connection PRB-22 (0.25 g, 0,485 mmol) was dissolved in absolute EtOH (5 ml) and the temperature is brought to 0°C in an inert atmosphere. Was added NaOEt (1M in ethanol, 0.54 ml, 0.54 mmol) and the mixture was stirred at 0°C for 30 minutes and the mixture was subjected to distribution between water and heptane. The aqueous layer was extracted with heptane and the combined organic layer was washed with 0.1 M HCl and dried. Cleaning method flash chromatography gave the output (13%) of the named compound PRB-23 as a colourless oil.

1H-NMR (200 MHz; CDCl3) δ of 0.8-1.0 (m, 6H), 1,2-1,4 (m, 4H), of 1.5-1.7 (m, 2H), 2,12 (m, 2H), 2,3-2,5 (m, 2H), 2.8 to 3.0 (m, 10H), 4,18 (t, 2H), 5,3-5,6 (m, 12H);

MS (electrospray ionization); 407 [M+Na].

[α]D-1.3° (C=1.00 m ethanol).

Obtaining the ethyl ester of α-phthalimide-D (PRB-24)

A mixture of ethyl ester of α-hydroxy-DHA (PRB-12) (373,5 mg, 1.00 mmol), phthalimide (178 mg, to 1.21 mmol) and triphenylphosphine (313,9 mg, 1.20 mmol) in THF (10 ml)was cooled to 0°C in an atmosphere of N2then drops was added diisopropylcarbodiimide (0,24 ml of 1.24 mmol). Tub with ice was removed and the reaction mixture was stirred at room temperature for 18 hours, after which it was implemented evaporation in a vacuum and were cleaned by the method of flash-chromatography on silica gel using as eluent a mixture of heptane/ethyl acetate (99:1-95:1), the resulting 323 mg (64%) of product as a yellow liquid.

1H NMR (200 MHz, CDCl3) δ of 0.95 (t, J=7.5 Hz, 3H), 1,22 (t, J=7,1 Hz, 3H), of 2.05 (m, 2H), 2,72-2,84 (m, 11 H), 3,02-3,22 (1H), 4,20 (q, 7=7,1 Hz, 2H), 4,87 (dd, J=11 Hz, J=4.9 Hz, 1H), 5,17-of 5.40 (m, 12H), 7,68 to 7.75 (m, 2H), 7,79 is 7.85 (m, 2H)

13With NMR (50 MHz, CDCl3) δ 14,0, 14,1, 20,4, 25,4, 25,4, 25,5, 27,0, 51,8, 61,7, 123,8, 124,3, 126,9, 127,5, 127,7, 127,9, 127,9, 128,1, 128,1, 128,3, 128,4, 131,6, 131,8, 131,8, 134,0, 167,3, 168,7 (2 hidden signal)

MS (electrospray ionization); 502 [M+H]+, 524 [M+Na]+

Obtaining the ethyl ester of α-acylamino-D (PRB-25) and ethyl ester of α-diethylamino-D (PRB-26)

To a mixture of ethyl ester of α-amino-DHA (PRB-17) (746,5 mg for 2.01 mmol), LiOH·H2O (171,6 mg, 4.09 to mmol) and molsieve 4A (599 mg) in DMF (4 ml) was added ethylbromide (3.0 ml, with 40.2 mmol), the mixture was stirred at room temperature for 71 hours. The mixture was diluted with diethyl ether (100 ml) and filtered. The organic phase was washed with 1 M NaOH (20 ml) and brine rastvorom (20 ml), dried (Na2SO4), filtered and evaporated in vacuo, purification was carried out according to the method of the flash-chromatography on silica gel, elution was carried out using heptane: ethyl acetate (95:5) - CH2CL2:2M NH3in methanol (99:1), the resulting 458 mg (53%) PRB-26 in the form of a yellow liquid and 152 mg (19%) PRB-25 as a yellow liquid.

PRB-26:

1H NMR (200 MHz, CDCl3) δ of 0.89 (t, J=7.5 Hz, 3H), of 1.03 (t, 3H), 1,24 (t, J=7,1 Hz, 6H), was 2.05 (quintet, J=7,1 Hz, 2H), 2,52 (m, 4H), was 2.76-to 2.85 (m, 12 H), the 3.35 (t, 1H), 4,13 (q, J=7,1 Hz, 2 H), 5,28-5,44 (m, 12 H)

13With NMR (75 MHz, CDCl3) δ 14,1, 14,3, 14,4, 20,5, 22,6, 25,5, 25,6, 25,7, 31,9, 44,4, 60,1, 62,9, 127,0, 127,8, 128,05, 128,13, 128,17, 128,22, 128,5, 132,0, 173,3 (5 hidden signals)

Examples

Models and methods used in the present invention to demonstrate the effects on the metabolic syndrome and type 2 diabetes type presented on the figure 2: Five blocks of experiments was performed to clarify, reduce whether derived DHA insulin resistance and/or ameliorate symptoms of the metabolic syndrome. The present invention is not limited pre is set implementation options and examples.

Example 1. Analysis of intracellular free fatty acids (free fatty acids) in the liver cells (block 1 in figure 2)

Prerequisites

In the first block of experiments (see figure 2) liver tissue from animals treated with PRB-1, 2, 5 and 7, were analyzed for the content of nonesterified free fatty acids. Animals were recruited from the fifteenth block experiments (pharmacodynamic effects derived DHA on the animal model of the metabolic syndrome). Animals received DHA (15% fat content of the diet) or derivatives of DHA (1,5% fat content of the diet) for 8 weeks and it was assumed that the intracellular concentrations of DHA and its derivatives DHA stable. The liver tissue was chosen for the reason that liver is the metabolic rate is very high.

Way

The liver samples were homogenized in cold phosphate buffer solution (PBS) and immediately extracted with chloroform: methanol (2:1)containing bottled hydroxytoluene (EIT), 0.2 mm, when using CIS-10-heptadecanoic acid as internal standard. The organic phases were dried over nitrogen, re-dissolved in acetonitrile with 0.1% acetic acid and 10 μm BHT for analysis by high performance liquid chromatography reverse phase (RP-HPLC-MS/MS). Total protein content was measured with the aid of the rd methodology Bio-Rad after homogenization.

System Agilent 1100 was used for reverse-phase column (column Supelco Ascentis C1S, 25 cm × 4.6 mm, inner diameter 5 μm) branch of DHA and its derivatives PRB for 22 minutes. The mobile phase - acetonitrile-water (87+13), containing 0.1% acetic acid. The column temperature was 35°C. For qualitative and quantitative analysis of the eluate was used for mass spectrometric detector "ABI Qtrap-400 three-stage quadrupole mass filter (ion trap) in the monitoring mode multiple ions. Ion pairs were as follows: 327,3/327,3 (DHA), 341,3/341,3 (PRB-1), 355,3/355,3 (PRB-2 and PRB-5), 387,3/387,3 (PRB-7), 267,2/267,2 (I.S. FA 17:1), respectively, under a single resolution. The exposure time was 100 MS, with the exception of FA 17:1, where the exposure time was 200 MS. Accurate verification of isomeric compounds PRB was obtained by the combination of retention time and the relationship of the mass/charge. A standard curve quadratic regression was used for the quantitative analysis after determining using an internal standard.

Results

The concentration of various derivatives of DHA and the DHA concentration were presented as μg per g of the total quantity of protein in liver cells. Figure 3 shows the concentrations of various PRBs from animals that received PRB-1, 2, 5 and 7 at a concentration of 1.5% of the total fat content in the diet with high fat content.

The highest vnutri etona concentration was obtained in the case of PRB-2. High intracellular content was observed in the case of PRB-5, although not to the same extent as in the case of PRB-2. This result is unexpected.

Figure 4 shows the intracellular concentration of DHA in liver tissues from animals that received different PRBs. The content of DHA was reached much higher levels in animals that received PRB-7 compared with the other three derivatives of DHA. In animals that received PRB-2, was observed the lowest concentration of DHA. Apparently, PRB-7 to some extent converted back into DHA. The highest intracellular concentration was observed in the case of PRB-2. This means that the connection PRB-2 will be more available as a ligand for nuclear receptors, thus, it will be observed therapeutic effects through controlling the level of blood glucose and lipid levels in the blood.

Example 2

Computer testing affinity (block 2 on figure 2)

Prerequisites

Nuclear receptors were sequenced and known amino acid sequence for PPARs and other receptors that play a role in the genetic control of glucose and fats. You can use rentgenocraniology and NMR spectroscopy PPAR and to evaluate the kinetics of binding can be used computerized testing affinity of fatty acids to R is the receptors. The geometric shape of the binding, often called methods or provisions binding, include the position of the ligand relative to the receptor and conformational regulation of ligand and receptor. Thus, can be analyzed effective docking of the ligand.

The affinity of the ligand relative to the receptor is determined using two different parameters: the docking of the ligand (derived D) into the binding site of the receptor and the electrostatic binding between some amino acids of the receptor and a carboxyl group or side chain "head" fatty acids (Krumrine).

As is known, the receptor PPARα is more "promiscuous" compared to PPARγ, PPARα is going to take more fatty acids as ligands compared to PPARγ. However, because patients with metabolic syndrome or type 2 diabetes are usually obese or are overweight and have an increased level of lipids in the blood, mainly elevated triglyceride and low cholesterol high density lipoprotein, is an important activating receptor PPARα. The ideal drug for the treatment of metabolic syndrome will act as a ligand for both receptors, preferably with a higher affinity to the receptor PPARγ.

Way

Classification according to which cnyh derived DHA according to their affinity of binding was calculated and presented, as most low affinity binding (LBA) and the average affinity binding (ABE).

All 15 derivatives of DHA (PRB-1-PRB-15) were tested using a computerized methods of docking. Some derivatives, such as PRB-1, PRB-2, PRB-7, PRB-9, PRB-10, PRB-11, PRB-12, PRB-13, PRB-14 and PRB-15 is represented as g and s enantiomers and in this case both were tested. PPARγ ligands rosiglitazone and pioglitazone, both in the r and s form, were tested for comparison. These compounds are registered as a pharmaceutical for the treatment of diabetes.

Results

The results are shown in table 1, which presents the parameters of the low binding energy of a single confirmation (LBE), the average binding energy (ABE) is correctly oriented ICM least 20 confirmed energy (found) of the tested compounds. It was also investigated the affinity for RXRα. Receptor RXRα interacts with the PPAR receptor, forming heterodimer when linking fatty acids.

Figure 5 shows the affinity of binding to receptor PPARγ, which mainly plays a role in the transcription of proteins involved in the control of glucose levels in the blood. Undoubtedly, PRB-2, both in r and s stereoisomeric form has a strong affinity to the receptor PPARγ. PRB-5 has a lower affinity, and PRB-8 has the highest ABE. These data are in a high degree of Noida is generated and can be expressed in a more efficient transcription activated PPARγ gene, responsible for monitoring glucose levels in the blood.

Figure 6 shows the affinity to the nuclear receptor PPARα, which is primarily responsible for metabolizing fat, blood lipids, biology of adipose tissue and weight control. Several derivatives of DHA are high affinity binding, but the highest affinity binding observed in the case PRB8. It is also a highly unexpected result.

Figure 7 shows the affinity to the nuclear receptor RXRα. The physiological result of binding with receptor RXRα just has not been. It is known that RXR binds to the PPAR receptors, thus forming heterodimer, which later triggers the transcription of the gene described.

ND=no docking, with a=double bond in fully-CIS form. r=R enantiomer, s=S enantiomer. ROSI=Rosiglitazone, RIO=Pioglitazone

Some PRBs have high LBE and ABE for receptors PPARα and PPARγ, even compared to the reference compound DHA, but also in comparison with PPARγ ligands by rosiglitazone and pioglitazone, both in r and in s form. This is an interesting observation indicates that some PRBs can be competitors well known antidiabetic medicines to rosiglitazone and pioglitazone.

Ethyl derivatives in the alpha position of these same girn the x acids, as in the r and s form, did not increase the degree of affinity. This was especially confirmed for receptor PPARγ. As already mentioned, the receptor PPARα is more "promiscuous" (contact can many fatty acids). In conclusion: many of the tested derivatives D showed high affinity to receptors PPARα and PPARγ, while the affinity of binding is higher than in the case of rosiglitazone and pioglitazone.

Example 3

Study of the affinity in transfected cells (block 3 in figure 2)

Prerequisites

The release of luciferase correlates with the transcription of genes. The binding of ligand to the nuclear receptor, such as PPARγ induces the transcription of the corresponding gene, thus, there is a release of luciferase. This technique, therefore, provides a measure of the affinity of the ligand to the receptor, and activation of the responsible gene.

Way

Transient transfection of COS cells-I was performed in 6-hole tablets, as described by Graham and van der Eb (Graham). For full studies on transfection in each well was made 5 μg of reporter construct, 2.5 µg of pSV-β-galactosidase as an internal control, and 0.4 µg SG5-Rγ2. Cells were collected after 72 hours, luciferase activity was measured respectively Protocol (Promega). The values of luciferase activity were normalized to the values of the-galactosidase activity. Adipocytes were transfected with D11 differentiation when using 16 ál LipofectaminPlus, 4 μl of Lipofectamine (Life Technologies Inc.), 0.2 µl pSG5-PPARγ and 100 ng RTC Renilla luciferase as a control of transfection efficiency. After transfection cells were incubated for three hours in serum containing medium, and then incubated for 48 hours in the same medium containing the appropriate agents. Luciferase activity was measured in accordance with the manufacturer's recommendations (Dual Luciferase assay, Promega). All transfections were performed three times.

Fatty acids (BRL or DHA) and connection PRBs (initial solutions) were restorany in DMSO to a final concentration of 0.1 M. we Then conducted the dissolution of up to 10 mm in DMSO, stored, carried out in 1.5 ml tubes (homopolymer, plastic tubes), which are filled with argon and stored at -20°C. 10 μm compounds PRBs or fatty acid and DMSO (control) was added to the medium after 5 hours after transfection. Transfected cells were maintained in medium for 24 hours prior to lysis reporter lyse buffer. Linking PRBs or fatty acids with the LBD of PPAR activates the binding of GAL4 with UAS, which in turn stimulates the tk promoter to launch the expression of luciferase. Luciferase activity was measured using a luminometer (TD-20/20 luminometer; Turner Designs, Sunnycvale, CA) and values were normalized to the protein content.

Results

The figure 8 presents the release of luciferase transfected cells, treated with different PRBs. The results indicate that PRB-1, 2, 6, 7, and 14 have a greater effect on the release of luciferase compared to R-3, 5, 9, 10, 11, 12 and 16.

Example 4

Study of the affinity for binding to PRBs and DHA from prone to obesity in animals with metabolic syndrome (box 4 in figure 2)

Prerequisites

To study the affinity PRB-2, 5 and 8 to PPARγ compared to 97% DHA and antidiabetic compounds rosiglitazone used animal model of the metabolic syndrome (prone to obesity mouse strain C57BL/6J), a study of the affinity was carried out by measuring the release of luciferase from the fat cells taken from these animals. Animals (n=8 in each group) for 8 weeks were given a diet high in fats (fats make up 60% of total calories, the same diet was used in block 5). After that, the animals received PRBs in the dose of 1.5% of the total fat content in the diet in the next two weeks. The group of mice treated with rosiglitazone, received 100 mg/kg feed. The control group continued to receive either a diet with a high fat diet or standard diet. Figure 9 shows the plan of study.

Way

After slaughter animals adipose tissue (epididymal or on the skin) was purified from other structures and cut into-inch pieces. Adipose tissue was washed in 0.9% NaCl and processed in 5 ml Krebs-Ringer containing HEPES free fatty acid bovine serum albumin, 200 nm adenosine, 2 nm glucose and 260 U/ml collagenase, for 1.5 hours at 37°C in shaking water bath. After the destruction by collagenase adipocytes were separated from debris by filtration. Cells were then washed in a solution of Krebs-ringer containing HEPES, bovine serum albumin, free fatty acids, 200 nm adenosine, 2 nm glucose, after which the cells were kept on shaking water bath at 37°C for a maximum of 30 minutes prior to electroporation.

Isolated primary adipocytes were transfected by electroporation with the aim of measuring the activity of specific regulatory proliferation of elements in the structure of the peroxisome (PPRE). In this case, we have inserted plasmid, which encodes the cDNA gene for Firefly luciferase under the control of the PPRE of the gene acyl-COA-oxidase. In addition, cells were transfected with a plasmid containing the luciferase cDNA coral Renilla under the control of the constitutively active promoter. Induced PPRE activity of Firefly luciferase was normalized according to the luciferase Renilla luciferase, thus, were adjusted for potential differences in the number of transfected cells. To measure luciferase is ignal we used a Dual-Luciferase(R) Reporter assay System (Promega, USA.

Adipose tissue was enough to highlight the adipocytes for duplicates. Animals of each group were scored with 2-day intervals and for each diet group were obtained from 4 independent transfection.

Results

During the first 8 weeks in mice fed a diet with high fat diet (HF diet) (33,7% fat, w/w), was observed a gradual increase in body weight compared with mice in the control, which received normal diet (4,5% w/w). Over the next two weeks the animals fed a diet high in fat, and animals receiving a diet with high fat diet in combination with rosiglitazone continued to gain weight at approximately the same speed as before. In the case of receiving a diet high in fat with the addition of PRB-8 and PRB-5 speed gaining weight decreased. However, in case of receiving PRB-2 and DHA (5% w/w) weight gain is completely stopped, and the body weight was even lower (figure 10). Food consumption was monitored from time to time (4). In this respect, differences between groups fed a diet high in fat, and groups treated with compounds PRB, were noted.

In the case of receiving a diet with high fat diet in combination with rosiglitazone endogenous activity receptacula PPARγ was approximately 2 times higher than in the sun the x groups, receiving specific diet (11). In addition, these fat cells become more sensitive to further stimulation in vitro with 5 uM of rosiglitazone (stimulation at 5, 12 times) compared with the diet high in fat (stimulation at 1.5 times). This rosiglitazone-sensitizing effect was observed in the groups receiving PRB-2 and PRB-5 (stimulation 2.6 times).

The data obtained in this study clearly showed activity against nuclear receptors PPAR, in particular it concerns the impact on the body weight, the most noticeable results have been obtained with the groups receiving the connection PRB-2. Even animals treated with compounds PRB-5 and PRB-8, not so fast I gained weight, like animals in the group treated with only diet high in fat. Interestingly, animals treated with rosiglitazone, the body weight was increased to the same extent as in animals treated with only diet high in fat. This clearly demonstrates the negative effects of only PPARγ ligands, such as glitazone as there is a risk of increased body weight, even if insulin resistance is reduced. However, when it comes to the activation of PPARγ, in this experiment, measured as luciferase activity, the effect of rosiglitazone more pronounced than the effect of any of the compounds PRBs. Connection PRB-2 and PRB-5 were the more effective, than PRB-8 and DHA (Fig).

Example 5

Pharmacodynamic effects derived DHA in animal model of the metabolic syndrome (box 5 in figure 2)

Prerequisites

Animal model of the metabolic syndrome (mice prone to obesity, strain C57BL/6J) were used to demonstrate the effects on typical laboratory and pathological anatomical features characteristic of the metabolic syndrome. When obtaining a diet high in fat (60% fat) in animals evolved obesity significantly increased the insulin level in the plasma, impaired glucose tolerance, serum increased the level of triglycerides and nonesterified fatty acids were observed to have a fatty liver.

Example 5A

The effects of derivative DHA in mice prone to obesity during the 4-month experimental diets.

Way

All experiments were performed on male mice of the strain C57BL/6, postema C57BL/N (supplier: Charles River, Germany, n=160, experiments a-C, see below) or postema C57BL/6J (provider: the Jackson laboratory, Bar Harbor, ME, USA, n=32, experiment D). The total number of animals used was higher due to culling (n=170 and 36, respectively). In the latter case, the animals were bred for several generations (<20) in the Institute of physiology. In the beginning of the experiment the animal was 14 weeks and their weight was 23.6-27.1, a week n the ed the beginning of the study the animals were sorted into 8 subgroups depending on their body weight. It was scrapped about 5-10% of the animals with the smallest and largest weight, respectively. Animals excluded from the study at this stage were euthanized by a method of cervical dislocation. Full verification of the health status of the mice was performed by supplier Charles River and at the beginning of the studies, serologic tests were performed by the laboratory ANLAB (Prague, Czech Republic). In addition, regular health check-up were performed in special rooms for animals with 3-month intervals during the use of indicator animals were conducted serological studies (ANLAB). Studies have shown that animals were free from infections.

Diet

Animals received three types of experimental diets:

(i) a Mixed diet (ssniffRM-H from SSNIFF Spezialdieten Gmbh, Soest, Germany; see also http://ssniff.de), in which proteins, fats and carbohydrates were respectively 33, 9 and 58% of the energy.

(ii) a Diet high in fat (cHF diet), prepared in the laboratory conditions in which proteins, fats and carbohydrates was, respectively, 15, 59, and 26% of the energy in the diet was added to a well-characterized composition of fatty acids (most of the lipids were isolated from corn oil; see Ruzickova 2004).

(iii) a Diet high in fat, prepared in laboratory conditions, in which of 0.15, 0.5 and 1.5% fat (component RL of ulusnogo oil) were replaced with various compounds PRB (PRB1, PRB2, PRB5, PRB7 and PRB8) or DHA. All these compounds were in the form of ethyl esters obtained from Pronova Biocare in sealed containers. The chemical composition of the compounds PRB was unknown to laboratory workers conducting experiments (Institute of physiology. The Prague Academy Of Sciences, Czech Republic).

After receiving the connection PRB kept in the refrigerator in their original containers. The containers were opened immediately before preparation of the experimental diets. Diet was kept in plastic bags filled with nitrogen, and stored at -70°C in small portions, which was enough to feed animals during the week. Fresh standards were given a 2-day interval or daily.

The plan of study

The study included 4 of the experiment. In each experiment a different connection PRB (or DHA) was mixed with a diet high in fat in three different concentrations of 0.15, 0.5 and 1.5% of the total fat content). Each experiment included a subgroup of mice served as control. Mice were planted in cells 4 and received a mixed diet before 3 months of age, then animals (n=8-13) were randomly divided into groups that received different experimental diets. After two months of receipt of specific diets (5 months of age), animals were left without food during the night and the slow and test was performed glucose tolerance (intraperitoneal injection). Animals were scored in 7-months of age after 4 months of experimental diets.

Research options

The parameters of the study were as follows: weight gain (grams), area under the curve (AUC) in tests of tolerance to glucose (mmol × 180 min), the level of insulin in plasma (ng/ml), the level of triglycerides in serum (TAGs mmol/1) and free fatty acid (NEFA mmol/1).

On Fig shows a typical curve of glucose clearance from the blood before and after giving the animal connection, reducing insulin resistance. Reduction of the area under the curve indicates the level of glucose in the blood decreases faster due to reduced insulin resistance.

Results

The results are presented in tables 2, 3 and 4. (*=significant difference compared to diets high in fat (P<0.05).)

Table 2 shows the effects in animals that received the test compounds PRB at a concentration of 1.5% compared with animals that received a diet high in fat or 97% DHA. The rate of gain in weight was significantly reduced in animals treated with PRB-2, compared to animals that received a diet with a high fat diet (cHF). In this group was slightly reduced food consumption. The most pronounced reduction of the area under the curve test glucose tolerance was observed in that the e group and even in animals, treated with PRB-1. The insulin level in plasma was significantly lower in the group treated with PRB-2, compared with controls cHF, even if the animals treated with PRB-1 and PRB-5 was also observed some effect on this parameter. In the group treated with PRB-2, showed the highest decrease in triglycerides (TAGs) and nonesterified fatty acids (NEFA).

Table 3 shows the effects in animals with the lowest concentration of 0.5%, the tested compounds compared to animals fed a standard diet, a diet with high fat diet (cHF) or 97% DHA. Weight gain was slightly lower in animals treated with PRB-2 and PRB-5. AUC test glucose tolerance, and insulin levels in plasma were, however, significantly lower only in the group treated with PRB-2.

Table 4 presents the results of the diet with the lowest concentration of PRB, 0,15%. In this case, the differences were very small. Weight gain was slightly lower in the groups treated with PRB-1 and R-2, whereas the AUC was significantly lower only in the group treated with PRB-2. The insulin level in the plasma was lower in the groups treated with PRB-1, 2 and 7.

Table 2
The effect of derivative PRB after 4 months of a diet containing 1.5% of PRB
STDFPRB-1PRB-2PRB-5PRB-7DHA
Body weight (grams)32,4±0,749,6±0,644,0±1,5*30,1±1,1*46,3±1,645,9±1,1*47,1±0,7*
Weight gain (grams)7,8±,0,425,2±0,520,2±1,3*6,4±0,8*22,4+1,421,7±0,9*23,0±0,8*
Food consumption (g/mouse/day)3,64±0,042,70±0,022,64±0,032,38±0,05*2,62±0,022,68±0,032,63±0,02
AUC glucose (mm × in)1124±571625±151913±68*982±89*1264±192 1122±732132±288*
The fasting glucose (mg/DL)77±3145±7130±1495±6*136±12120±9138±7
Insulin (ng/ml)1,03±0,095,35±0,362,73±0,330,60±0,18*2,47±0,19*4,42±0,876,55±0,31
Triglycerides (mmol/l)1,41±0,091,45±0,071,58±0,080,71+0,01*1,19±,0,071,15±0,081,91±0,26*
The free fatty acid (nmol/l)0,57±0,050,61±0,040,63±0,03*0,54±0,03*0,72±0,050,82±0,060,98±0,07

Table 3
The effect of derivative PRB after 4 months of diet containing 0.5% PRB
STDcHFPRB-1PRB-2PRB-5PRB-7DHA
Body weight (grams)32,4±0,749,6±0,647,4±0,645,8±1,7of 45.7±1,548,8±0,946,9±0,7*
Weight gain (grams)7,8±0,425,2±0,523,8±0,5of 21.9±0,622,0±1,424,8±0,822,9±0,7*
Food consumption (g/mouse/day)3,64±0,042,70±0,022,67±0,042,69±0,042,63±0,022,69±0,032,70±0,03
AUC glucose (mm x 180 min)1124+571625±151 1596+2051224±72*1581±2311674±2031816±182
The fasting glucose (mg/DL)77±3145±7131±7136±7130±7152±6136±8
Insulin (ng/ml)1,03±0,085,35±0,363,93±0,592,75±0,21*5,12±0,934,10±0,57*of 5.82±0,47
Triglycerides (mmol/l)1,41±0,091,45±0,072,03±0,221,29±0,081,4B±0,171,42±0,081,78±0,08*
The free fatty acid (nmol/l)0,57±0,050,61±0,040,73±0,04*0,75±0,040,77±0,03*0,87±0,040,89±0,03

Table 4
The effect of derivative PRB after 4 months of a diet containing 0.15% of PRB
STDcHFPRB-1PRB-2PRB-5PRB-7DHA
Body weight (grams)32,4±0,749,6±0,647,2±1,346,7±1,148,0±0,847,4±0,8*48,3±0,6
Weight gain (grams)7,8±0,425,2±0,522,9±1,122,8±0,924,2±0,523,2±0,7*24,3±0,8
Food consumption (g/mouse/day)3,64±0,042,70±0,022,63±0,04to 2.57±0,03*2,66±0,022,59±0,022,79±0,03
AUC glucose (m is × 180 min) 1124±571625±1511291±1721071±148*1443±701425±971477±214
The fasting glucose (mg/DL)77+3145±7126±15132±5151+5141±9141±10
Insulin (ng/ml)1,03±0,085,35±0,363,50±0,294,00±0,646,21±0,453,76±0,72*or 4.31±0,39*
Triglycerides (mmol/l)1,41±0,091,45±0,071,75±0,081,42±0,071,B4±0,281,41±0,111,50±0,13
The free fatty acid (nmol/l)0,57±0,050,61±0,040,62±0,04*0,78±0,04*0,71±0,090,96±0,07

In conclusion: the study of the application of PRB-1, 2, 5 and 7 within 4 months we are likely to stay obese animals with insulin resistance and metabolic syndrome showed obvious and unexpected effect of tested compounds PBRs and in particular derived DHA connection PRB-2 on insulin resistance and symptoms of metabolic syndrome, there was a decrease weight, decrease in AUC in the intraperitoneal test, glucose tolerance, and reduced levels of triglycerides and nonesterified fatty acids. Effects were observed in the group that received a dose of PRB 1.5%, and the group treated with the dose of 0.5%. Some effects were noted even in the group treated with the lowest concentration of 0.15%.

The connection test PRB-8 started later, so in the three groups receiving diets containing concentrations of this compound 1,5%, 0,5% and 0.15%, data were obtained only 2 months of research. In the group receiving 1.5%), body weight (BW) was 28.0±0.7 g compared with control 29.6±0.9 AUC was 1031±104 compared with control 1074±91. These differences are small, but the trend is interesting. For these parameters there were no differences between the experimental diets, in which animals received the lowest dose of 0.5% and 0.15%, and control groups. Data obtained with PRB-8 (use within 2 months the Germans), showed that there is a tendency to reduce the weight and AUC.

Example 5b

The effect of derivative DHA on metabolic syndrome and insulin resistance

Way

In another experiment PRB-2, PRB-5 and PRB-7 was tested on animals of the same breed. In this experiment, the animals were first given a diet high in fat (like in the previous experiment 5A) within 8 weeks, the animals developed insulin resistance and metabolic syndrome, and then they received connection PRBs. The initial dose of PRBs 15% fat content, but animals are not tolerated this dose. After 2-week period, the animals received 1.5% of the PRB-2, 5% and 1.5% PRB-5 and 1.5% and 0.5% PRB-7.

Results

The weight loss was very pronounced in animals treated with PRB-2. Even in animals treated with PRB-5, there was a slight decrease in weight, but in a higher dose of 5%. Triglyceride levels decreased in all tested derivatives compared to control animals receiving a diet with a high fat diet (cHF diet). Reduction of the level of nonesterified fatty acids was more pronounced when using PRB-2 and PRB-5, but in the different dose (see Fig).

The level of cholesterol in the blood were reduced in animals treated with PRB-2 and PRB-5. The level of blood glucose was not changed due to the fact that these animals are in a pre-diabetic condition is a normal glucose level because of the high level of secretion of the production of insulin. However, the most important is the fact that the level of insulin in plasma was significantly reduced in the group of animals treated with PRB-2 in much lower concentrations compared with the second very effective DHA derived PRB-5. Even when using PRB-7 was observed effects on the concentration of insulin (see Fig).

When using PRB-2 was observed a statistically significant decrease in the AUC of glucose in blood at all time points compared with baseline values. This means that the level of glucose in the blood decreased after application of PRB-2 at a concentration of 1.5%. When using PRB-5 and PRB-7 some effect was observed, but to a lesser extent than in the case of PRB-2 (see Fig).

These results are very unexpected and speak about the positive effect of these compounds in the metabolic syndrome and type 2 diabetes type. Almost all of these patients are obese or are overweight and the results showed that the drug is promising in reducing weight. The most applicable tools for the treatment of type 2 diabetes type today are thiazolidinone that are strong ligands of PPARγ, thus reducing insulin resistance, which often leads to increased body weight, which is very undesirable for these patients (Yki-Jarvinen 2004).

Reducing the level of triglycerides in the serum of the AOC and is another very important effect, which was shown in these experiments. In patients with metabolic syndrome and type 2 diabetes type often increased the level of triglycerides. The effects of derivative DHA to reduce the level of triglycerides are very positive and again the connection PRB-2 showed the greatest effect at the lowest doses. Very positive effects on insulin levels in plasma and test glucose tolerance are very promising and very unexpected. Taken together, the results obtained with derivatives of DHA, especially with PRB-2, are very encouraging for forming a good base for the development of antidiabetic drugs.

Example 5C

The use of derivatives of DHA in fatty degeneration of the liver

Way

In experiments with derivatives of DHA were selected tissue samples was conducted histological examination. After waxing is produced tissue samples of liver, adipose tissue, skeletal muscle, pancreas, kidneys were stained with eosin-hematoxylin.

Results

Pathological changes in tissues except liver tissue were detected. Animals in the control, which received a diet with high fat content, was observed fatty degeneration of the liver (hepatic steatosis). Fat droplets in the liver were easily distinguishable from normal cells PE the Yeni. In animals treated with compounds of PRB-1, 5, and 7, it was noted the low degree of fatty degeneration of the liver. However, in animals treated with PRB-2 at a concentration of 1.5%was not observed any signs of fatty degeneration of the liver. This observation is extremely important and is of great importance for the treatment of patients with insulin resistance, obesity and type 2 diabetes type. Fatty degeneration of the liver is very common in these patients, which is associated with excess fatty acids and triglycerides, biological markers in the development of insulin resistance and metabolic syndrome. Derived DHA reduce fatty degeneration of the liver, the most pronounced effect showed the connection PRB-2.

DISCUSSION AND CONCLUSION

In this application is considered a new group of compounds that activate nuclear receptors, especially PPARγ and PPARα, thus, providing a range of therapeutic effects in the treatment of insulin resistance, metabolic syndrome, type 2 diabetes type, cardiovascular disease and other diseases associated with atherosclerosis.

Members of this group are derivatives of DHA with different side chains in the alpha position of the molecule. A large number of alpha-substituted derivatives of DHA have been tested and compared with controls, and with pure DHA and EPA. Some the tested compounds showed interesting biological effects, relevant to the creation of anti-diabetic drugs based on them.

Interesting and yet confusing is the fact that the ethyl ester of alpha-ethyl-DHA (PRB-2) was significantly more effective in some of the tests used to demonstrate the effects associated with insulin resistance and due to this disease, which is caused by this pathophysiological condition, these diseases are metabolic syndrome, type 2 diabetes type, cardiovascular disease and other diseases associated with atherosclerosis. The liver tissue of animals receiving various derivatives of DHA (block 1), was saturated ethyl ester, alpha-ethyl-DHA, this indicates that this compound has not been used for the synthesis of triglycerides, eicosanoids, or other intermediates of metabolism. Indirectly this means that the alpha-ethyl-DHA will be available for binding to nuclear receptors such as PPARs.

Testing of affinity to PPARγ and PPARα when using docking technology showed that a large number of derivatives of DHA have affinity to both receptors and last but not least to PPARγ, which is probably the most important nuclear receptor involved in the activation of genes responsible for metabolizing glucose in the blood. Especially alpha-ethyl-DHA (PRB-2) and alpha-DHA (PRB-8) had a strong affinity for these nuclear receptors. Compared to alpha-diethyl-DHA alpha-ethyl-DHA has two stereoisomer, r and s forms. When using the docking technology has been shown that both stereoisomer have the same affinity for PPARγ and PPARα, this means that neither r nor s form does not have advantages over racemic form. Moreover, the racemic form may have advantages over each of the isomers.

When affinity was studied in transfected cells carrying nuclear receptor and item response DNA, it was shown that some compounds PRBs have a strong affinity, measured as the release of luciferase. The best results were observed in the case of alpha-ethyl-DHA (PRB-2) and PRB-6, 7, and 14.

Five derivatives of DHA was tested on mice of the strain C57BL/6, have developed insulin resistance and metabolic syndrome when receiving a diet high in fat. The compound alpha-ethyl-DHA (PRB-2) was tested in three separate experiments, whereas PRB-1, 5, and 7 were tested in two experiments and the compound alpha-diethyl-DHA (PRB-8) was tested in one experiment. All derivatives showed significant biological effects. However, in the case of alpha-ethyl-DHA (PRB-2) was observed most promising effects: decrease of body weight, AUC in intraperitoneal test glucose tolerance, insulin levels in plasma and decrease the con is entrale triglycerides and nonesterified fatty acids in the serum. These results were obtained with doses of 1.5% and 0.5%. At the lowest dose of 0,15% of the effects were not clearly expressed. The compound alpha-ethyl-DHA (PRB-2) in the dose of 1.5% was also effective in the prevention of fatty degeneration of the liver, which often develops in people and animals with insulin resistance and metabolic syndrome.

The compound alpha-ethyl-DHA (PRB-2) acts 10-30 times stronger than pure DHA. Comparison results derived DHA with the original molecule DHA potency was highly unexpected.

As alpha-ethyl-DHA (PRB-2) apparently works by binding to nuclear receptors PPARγ and PPARα, the connection not only has an interesting therapeutic effects on the metabolism of carbohydrates and fats, not least in patients with insulin resistance, metabolic syndrome and diabetes of the 2nd type, but also reduces the weight and have an overall anti-inflammatory effect. Directly or through the influence factry risk compound alpha-ethyl-DHA (PRB-2) can prevent the development of cardiovascular diseases such as myocardial infarction and ischemic stroke, and this compound can reduce the mortality rate from cardiovascular diseases.

The pharmaceutical agents which act as ligands of PPARγ, are already present in the pharmaceutical market, but even if e and compounds have a positive effect on carbohydrate metabolism, they are characterized by side effects such as elevated triglycerides, increased body weight and swelling. Provided in this application alpha-substituted derivatives of DHA exert a combined effect on the receptor PPARγ and PPARα, which may be an advantage for patients with insulin resistance, metabolic syndrome and type 2 diabetes type. In addition, the combined influence affects the level of blood lipids, inflammation, atherosclerosis and, consequently, on cardiovascular diseases.

The present invention is not limited to the shown variants of implementation and examples.

Sources of information

Simonopoulos AR. Essential fatty in health and chronic disease. Am J ClinNutr 1999; 70 (Suppl):560S-569S.

Geleijnse JM, Giltay EJ, Grobbee DE, et al. Blood pressure response to fish oil supplementation: metaregression analysis of randomized trials. J Hypertension 2002; 20:1493-1499.

Storlien LH, Hulbert AJ, Else and PL. Polyunsaturated fatty acids, membrane function and metabolic diseases such as diabetes and obesity. Curr Opin Clin Nutr Metab Care 1998; 1 - 559-563.

Jump DB. The biochemistry of n-3 polyunsaturated fatty acids, J Biol Chem 2002; 277 - 8755-8758.

Pawar A and D. Jump Unsaturated fatty acid regulation ofperoxisomes proliferator-activated receptor alfa activity in rat primary hepatocytes. J Biol Chem 2003; 278:35931-35939.

Meigs JB, Wilson PWF, Nathan DM, et al. Prevalence and characteristics of the metabolic syndrome in the San Antonio Heart and Framingham offspring studies. Diabetes 2003; 52:2160-2167.

Storlien LH, KraegenWE, Chisholm DJ, et al. Fish oil prevents insulin resistance induced by high fat feeding in rats. Science 1987; 237:885-888.

Field CJ, Ryan EA, Thomson ABR et al. Diet fat composition alters membrane phosphoipid composition, insulin binding and glucose metabolism in adipocytes from control and diabetic animals. J Biol Chemistry 1990; 265:11143-11150.

Yki-Jarvinen, H. Thiazolidinediones. NEJM 2004; 351:1106-1118. Adams M, Montague CT5 Prins JB, et al. Activators of peroxisomes proliferator - activated receptor gamma have depot-specific effects on human preadipocyte differentiation. J Clin Invest 1997, 100:3149-3153.

Ruzickovaj, Rossmeisl M, Prazak T, et al. Omega-3 PUFA of marine origin limit diet-induced obesity in mice by reducing cellularity of adipose tissue, Lipids 2004; 39: 1177-1185.

Vaagenes H, Madsen L, Dyroy E, et al. The hypolipidaemic effect of EPA is potentiated by 2 - and 3-methylation. Biochim Pharmacol 1999; 58:1133-1143.

Larsen L, Granslund L, Holmeide AK, et al. Sulfur-substituted and [alpha]-methylated fatty acids as peroxisome proliferator-activated receptor activators. Lipids 2005, 40:49-57.

Larsen L, Horvik K, Sorensen HIN, et al. Polyunsaturated thia - and oxa-fatty acids: incorporation into cell-lipids and their effects on arachidonic acid - and eikosanoids synthesis. Biochim et Biophys Acta 1997, 1348:346-354.

Larsen, et.al. Biochemical Pharmacology 1998; 55, 405.

Chih-Hao L, Olson P, and Evans RM. Lipid metabolism, metabolic diseases, and peroxisome proliferator-activated receptors. Endocrinology 2003; 144:2201-2207.

Willumsen N, Waagenes H, Holmsen H, et al. On the effect of 2-deuterium - and 2-methyl-eicosapentaenoic acid derivatives on triglycerides, peroxisomal beta-oxidation and platelet aggregation in rats. Biochim Biophys Acta 1998; 1369:193-203.

Mitsunobu O, Synthesis 1981;1.

Ager DJ Prakash I, and Schaad DR. 1,2-amino alcohols and their heterocyclic derivatives as chiral auxiliaries in asymmetric synthesis Chem Rev 1996; 96:835-876.

1. The compound of the formula

(4Z, 7Z, 10Z, 13Z, 16Z, 19Z)-ethyl 2-Aceldama-4,7,10,13,16,19-hexanoate.

2. The compound according to claim 1, characterized in that the compound is present as the S enantiomer.

3. The compound according to claim 1, characterized in that the compound is present as the R enantiomer.

4. Connect the tion according to claim 1, characterized in that the compound is present as a mixture of R and S enantiomers.

5. The compound according to claim 4, characterized in that the mixture is racemic.

6. Composition for the treatment of diabetes, to reduce insulin levels, glucose levels in the blood level of triglyceride in the serum for the treatment of dyslipidemia to reduce cholesterol levels in the blood, to reduce body weight and for the treatment of peripheral resistance to insulin, comprising an effective amount of a compound according to claim 1 and at least one pharmaceutically acceptable excipient.

7. The composition according to claim 6, formulated for oral administration.

8. The composition according to claim 6 in the form of a capsule or sachet-powder.

9. The composition according to claim 6, formulated for intravenous, subcutaneous or intramuscular injection.

10. The composition according to claim 6, composed in such a way that it delivers a daily dose of from 10 mg to 10 g

11. The composition of claim 10, composed in such a way that it delivers a daily dose of from 100 mg to 1 g of the specified connection.

12. Dietary lipid composition, comprising a pharmaceutically effective amount of a compound according to claim 1.

13. Dietary lipid composition according to item 12, characterized in that the compound according to claim 1 is at least 60% of the total weight of the composition.

14. Dietary lipid composition according to item 13, wherein the connection p of the claim 1 is at least 90% of the total weight of the composition.

15. Dietary lipid composition according to item 12, characterized in that the composition further includes a pharmaceutically acceptable antioxidant.

16. Dietary lipid composition according to item 15, wherein the pharmaceutically acceptable antioxidant is tocopherol.

17. A method of treating and/or preventing diabetes comprising the administration to a human or animal a pharmaceutically effective amount of a compound according to claim 1.

18. The method according to 17, wherein the diabetes is type 2 diabetes type.

19. The way to reduce insulin levels, glucose levels in the blood and/or level of triglyceride in serum, comprising the administration to a human or animal a pharmaceutically effective amount of a compound according to claim 1.

20. The method of treatment and/or prevention of dyslipidemia, comprising the administration to a human or animal a pharmaceutically effective amount of a compound according to claim 1.

21. The method according to claim 20, characterized in that dyslipidemia is gipolipidemicheskoe state.

22. The method according to item 21, wherein hyperlipidemics state is hypertriglyceridemia.

23. The way to reduce cholesterol in the blood, comprising the administration to a human or animal a pharmaceutically effective amount of a compound according to claim 1.

24. The way to reduce body weight and/or prevent gaining weight, R is the overall introduction to the human or animal a pharmaceutically effective amount of a compound according to claim 1.

25. A method of treating or preventing peripheral resistance to insulin, comprising the administration to a human or animal a pharmaceutically effective amount of a compound according to claim 1.

26. The compound of formula (I)

wherein X is a carboxylic acid, carboxylate, carboxylic anhydride, diglyceride, triglyceride, phospholipid or carboxamido,
or any pharmaceutical acceptable salt.

27. Connection p formula (II)

28. Connection p in salt form, wherein X is COO-Z+where Z+is Li+, Na+, K+, NH4+or substituted NH4+.

29. Connection p in the form of a salt of the formula

where Z2+represents Mg2+or Sa2+.

30. Connection p, characterized in that the carboxylate group is selected from ethylcarboxylate, methylcarbamate, n-propellerblade, isopropylcarbonate, n-butylcarbamoyl, sec-butylcarbamoyl and n-lexiscanlexiscan.

31. Connection p, characterized in that carboxamidine group selected from primary carboxamide, N-methylcarbamyl, N,N-dimethylcarbamate, N-ethylcarbodiimide and N,N-diethylbenzamide.

32. Connection p, differently the, what carboxylate group is methyl or ethylcarboxylate.

33. Connection p, characterized in that the compound is present as the S enantiomer.

34. Connection p, characterized in that the compound is present as the R enantiomer.

35. Connection p, characterized in that the compound is present as a mixture of R and S enantiomers.

36. Connection p, characterized in that the mixture is racemic.

37. Composition for the treatment of diabetes, to reduce insulin levels, glucose levels in the blood level of triglyceride in the serum for the treatment of dyslipidemia to reduce cholesterol levels in the blood, to reduce body weight and for the treatment of peripheral resistance to insulin, comprising an effective amount of a compound according p and at least one pharmaceutically acceptable excipient.

38. The composition according to clause 37, additionally comprising the compound of the formula

39. A method of treating and/or preventing the disease or condition is selected from the peripheral resistance to insulin/diabetic condition; and the condition of obesity or overweight; including the introduction of the human or animal a pharmaceutically effective amount of a compound according p.

40. The method according to § 39, wherein the diabetic condition is diabetes of the 2nd type.

41. With whom persons treatment and/or prevention of dyslipidemia, includes introduction to the human or animal a pharmaceutically effective amount of a compound according p.

42. The method according to paragraph 41, wherein the dyslipidemia is gipolipidemicheskoe state.

43. The method according to § 42, wherein the dyslipidemia comprises elevated levels of triglyceride and/or non-HDL (LDL cholesterol and VLDL cholesterol).

44. The way to reduce insulin levels, glucose levels in the blood and/or level of triglyceride in the serum by introducing the human or animal a pharmaceutically effective amount of a compound according p.

45. The way to reduce body weight and/or prevent gaining weight through the introduction of the human or animal a pharmaceutically effective amount of a compound according p.

46. The method of activation and/or binding isoforms of at least one of the human receptor, activating peroxisome proliferation (PPAR) by introducing the human or animal a pharmaceutically effective amount of a compound according p.

47. The pharmaceutical composition according to clause 37, prepared for oral administration.

48. The pharmaceutical composition according to clause 37 in the form of a capsule or sachet-powder.

49. The pharmaceutical composition according to clause 37, prepared for intravenous, subcutaneous or intramuscular injection.

50. The pharmaceutical composition according to clause 37, is provided so she delivers a daily dose of from 10 mg to 10 g

51. The pharmaceutical composition according to clause 37, is made in such a way that delivers a daily dose of from 100 mg to 1 g

52. Dietary lipid composition, comprising a pharmaceutically effective amount of a compound according p and at least one pharmaceutically acceptable excipient.

53. Dietary lipid composition according to paragraph 52, wherein the compound of formula (I) is at least 60% of the total weight of the composition.

54. Dietary lipid composition according to item 53, wherein the compound of formula (I) is at least 90% of the total weight of the composition.

55. Dietary lipid composition according to paragraph 52, additionally comprising a pharmaceutically acceptable antioxidant.

56. Dietary lipid composition according to § 55, wherein the pharmaceutically acceptable antioxidant is tocopherol.

57. The method of obtaining compounds of ethyl (fully-Z)-2-Aceldama-4,7,10,13,16,19-hexanoate

including:
a) reaction of a solution containing ester of DHA, with a strong dinucleophiles the basis for obtaining enolate ether, and
b) response enolate obtained in stage a), with electrophilic ethyl reagent, representing ethyliodide;
c) extraction of the product from the solution obtained in stage b)with a solvent.

5. The method according to § 57, wherein the DHA ester is obtained from a source of vegetable, microbial and animal origin, or their combinations.

59. The method according to § 57, characterized in that the ester of DHA derived from marine grease.

60. The method according to p, wherein the marine oil is fish oil.

61. The method according to § 57, characterized in that the strong dinucleophiles base is selected from diisopropylamide lithium hexamethyldisilazane potassium and hexamethyldisilazane sodium.



 

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12 cl, 2 tbl, 12 ex

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28 cl, 17 tbl

FIELD: chemistry.

SUBSTANCE: method of regioselective obtainment of 1-R1-2-R2-3-acetyl-glycerol derivative of the Formula 1 involves the following stages: Obtainment of 1-R1-3-(protective group)-glycerol of Formula 3 by adding protective group to 3rd position in 1-R1-glycerol of Formula 2; obtainment of 1-R1-2-R2-3-(protective group)-glycerol of Formula 4 by adding R2 group to 2nd position of 1-R1-3-(protective group)-glycerol of Formula 3, where R2 group is added by reaction of R2-OH with 1-R1-3-(protective group)-glycerol in the presence of aprotic organic solvent, catalyst and dehydrating medium; aprotic organic solvent is selected out of group consisting of hexane, heptane, dichloromethane, ethyl acetate, tetrahydrofuran and mixes thereof; dimethylaminopyridine is catalyst; and dicyclohexylcarbodiimide is dehydration medium; simultaneous removal of protective group and acetylation of 1-R1-2-R2-3-(protective group)-glycerol of Formula 4, where protective group removal reaction and acetylation reaction are performed using Lewis acid and acetic anhydride or using acetylation agent; Lewis acid is selected out of group including zink chloride (ZnC2), tin chloride (SnCl2), boron trifluoride diethyl ether (BF3Et2O) and mixes thereof; acetylation agent is selected out of group including acetylchloride, acetylbromide and mixes thereof, where compounds of Formulae 1-4 are racemic or optically active; R1 is palmitic acid group, R2 is linoleic acid group; P is trityl or trialkylsilyl as protective group; alkyl in trialkylsilyl is an alkylic group containing 1-5 carbon atoms, so that if the protective group is trityl then 1-R1-3-(protective group)-glycerol is obtained in the presence of pyridine solvent at 40-60°C or in the presence of nonpolar aprotic organic solvent and organic base within 0°C to room temperature range; nonpolar aprotic organic solvent is selected out of group including pyridine, dichloromethane, tetrahydrofuran, ethyl acetate and mixes thereof; organic base is selected out of group including triethylamine, tributylamine, 1,8-diazabicyclo[5,4,0]-7-undecene (DBU) and mixes thereof, and if the protective group is trialkylsilyl then 1-R1-3-(protective group)-glycerol is obtained in the presence of aprotic organic solvent and organic base within 0°C to room temperature range; aprotic organic solvent is selected out of group including dichloromethane, tetrahydrofuran, ethyl acetate, dimethylformamide and mixes thereof; and organic base is selected out of group including imidazole, triethylamine, and mixes thereof. [Formula 1] , [Formula 2] , [Formula 3] , [Formula 4] .

EFFECT: obtainment of glycerol derivative with high efficiency and output.

8 cl, 10 ex

FIELD: chemistry.

SUBSTANCE: invention relates to novel hexafluoroisopropanol-substituted ether derivatives of formula (I) to their pharmaceutically acceptable salts and to esters which are capable of bonding with LXR-alpha and/or LXR-beta, as well as to pharmaceutical compositions based on said compounds. In formula (I) R1 is hydrogen, lower alkyl or halogen, one of groups R2 and R3 is hydrogen, lower alkyl or halogen, and the second of groups R2 and R3 is -O-CHR4-(CH2)m-(CHR5)n-R6. Values of R4, R5, R6 m and n are given in the formula of invention.

EFFECT: novel compounds have useful biological properties.

22 cl, 4 dwg, 102 ex

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention relates to novel compounds of the formula (I) and their pharmaceutically acceptable salts and esters. In the general formula (I) X means oxygen (O) or sulfur (S) atom; R means hydrogen atom (H) or (C1-C6)-alkyl; R1 means H, -COOR, (C3-C8)-cycloalkyl or (C1-C6)-alkyl, (C2-C6)-alkenyl or (C1-C6)-alkoxyl and each of them can be unsubstituted or comprises substitutes; values of radicals R2, R3, R4, R5 and R6 are given in the invention claim. Also, invention relates to a pharmaceutical composition based on compounds of the general formula (I) and to intermediate compounds of the general formula (II) and the general formula (III) that are used for synthesis of derivatives of indane acetic acid. Proposed compounds effect on the blood glucose level and serum triglycerides level and can be used in treatment of such diseases as diabetes mellitus, obesity, hyperlipidemia and atherosclerosis.

EFFECT: valuable medicinal properties of compounds and pharmaceutical composition.

28 cl, 6 tbl, 6 sch, 251 ex

The invention relates to an improved process for the preparation of ethyl ester of 10-(2,3,4-trimetoksi-6-were) decanoas acid, which is an intermediate product, suitable for the synthesis of idebenone - drug nootropic action

The invention relates to ester compounds, method of their production and their use as a means for spooling the fiber

The invention relates to organic chemistry, namely the method of obtaining the ethyl ester of 10-(2,3,4-trimetoksi-6-were) decanoas acid - intermediate, suitable for the synthesis of idebenone - drug nootropic action

The invention relates to compounds of the formula

< / BR>
in which R1and R2each independently represents CNS group containing 1 to 4 carbon atoms, R3- H or acylcarnitine group containing 2 to 5 carbon atoms, R4- CNS group containing 1 to 4 carbon atoms, in free form and also, if such exist, in the form of salt

FIELD: medicine.

SUBSTANCE: invention relates to applications of compound 11-deoxyprostaglandin of general formula (IV) for obtaining composition for treatment of central nervous system disorder and for obtaining composition for protection of endothelial cells of brain vessels, to pharmaceutical composition based on said compounds, to method of treating central nervous system disorder, as well as to method of treating central nervous system disorder, as well as to compounds of general formula (IV) or their pharmaceutically acceptable salts, esters or amides, on condition that compound is not 11-desoexy-13,14-dihydro-15-keto-16,16-difluor- PGE1. , where L represnts hydroxy, lower alkanoyloxy or oxo; A represents -COOH or its pharmaceutically acceptable salt, ester or amide; B represents -CH2-CH2 or -CH=CH-; Z represents , or , where R4 and R5 represent hydrogen or hydroxy. R4 and R5 cannot represent hydroxy simultaneously; X1 and X2 represent similar or different halogen atoms; R1 represents saturated or unsaturated bivalent lower or middle aliphatic hydrocarbon; R2 represents single bond or lower alkylene and R3 represents linear lower alkyl.

EFFECT: increase of treatment efficiency.

14 cl, 7 ex, 6 tbl, 20 dwg

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