KR101255650B1 - New DHA derivatives and their use as medicaments - Google Patents

Abstract

The present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt, solvate, complex, or prodrug thereof:

 Formula (I)]

The R ₁ And R ₂ is the same or different and is a hydrogen atom, a hydroxy group, an alkyl group, a halogen atom, an alkoxy group, an acyloxy group, an acyl group, an alkenyl group, an alkynyl group, an aryl group, an alkylthio group, an alkoxycarbonyl group, an alkylsulfinyl group, alkyl A sulfonyl group, an amino group, and an alkylamino group; X is a carboxylic acid group, a carboxylate group, or a carboxamide; And the compound of formula (I) is (all-Z) -4,7,10,13,16,19-DHA (docosahexaenoic acid), alpha-methyl DHA, alpha-methyl DHA methyl ester, alpha-methyl DHA ethyl It is not an ester or alpha-hydroxy DHA ethyl ester.

Fatty acid compositions and pharmaceutical compositions comprising such compounds are disclosed. Uses of such compounds are disclosed, in particular as medicaments for type 2 diabetes.

Description

New DHA derivatives and their use as medicaments

The present invention relates to compounds of the general formula (I) and to their use as medicaments, in particular, type 2 diabetes mellitus (diabetes mellitus, type 2) and the medicaments of the preceding stages:

(I)

The invention also relates to a pharmaceutical composition comprising said compound of formula (I) and a fatty acid composition comprising said compound of formula (I).

As the incidence of type 2 diabetes increases worldwide, extensive public health and medical tests are being conducted to successfully prevent and treat it. Overweight and obesity, which are strongly associated with type 2 diabetes, occur simultaneously to interfere with diabetes treatment, and to cause hypertension, dyslipidemia, and atherosclerosis related diseases Increase the likelihood.

The pathophysiological condition indicative of the development of type 2 diabetes is associated with a decrease in insulin effect in peripheral tissues, called insulin resistance. These tissues are mainly muscle, fat and liver. Muscle tissue is the major tissue involved in insulin resistance in type 2 diabetes. Syndromes due to insulin resistance, hypertension, lipid metabolism and systemic proinflammatory states are called metabolic syndrome. The incidence of metabolic syndrome among adult populations in developed countries is 22-39% (Meighs 2003).

At present, the most effective approach to alleviate and prevent metabolic syndrome is lifestyle control with weight loss, reduced saturated fatty acid intake, and increased physical activity with adequate drug therapy. Healthy diets to avoid excessive energy intake include replacing saturated fats with mono and polyunsaturated fatty acids. In particular, long-chain omega-3 fatty acids, e.g., eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), obtained from fatty fish, are used to prevent type 2 diabetes. It has been shown to be effective.

EPA and DHA have effects on physiological processes that affect normal health and chronic diseases such as plasma fat concentration control, cardiovascular and immune responses, insulin action and neurogenesis and visual function. There is solid evidence that these substances play a beneficial role in the prevention and management of coronary heart disease, lipid metabolism, type 2 diabetes, insulin resistance and hypertension (Simonopoulos 1999; Geleijnse 2002; Storlien 1998). . Recent studies have found that nuclear omega-3 fatty acids regulate nuclear receptors, such as peroxisome proliferator-activated receptors (PPARs), while regulating gene expression involved in fat and glucose metabolism and adipogenesis. It acts via transit and plays an important role in gene expression (Jump 2002). PPARs are nuclear fatty acid receptors that play an important role in metabolic diseases associated with obesity, such as hyperlipidemia, insulin resistance, and coronary heart disease.

Three subtypes of α, γ, and δ have distinct patterns of expression, releasing sensory components of different lipoproteins and regulating fat homeostasis, depending on the needs of a particular tissue. PPARα increases fatty acid degradation in the liver and is a molecular target of the lipid-lowering fibrates. PPARγ, on the other hand, is essential for adipocyte differentiation and modulates the activity of insulin-sensitive thiazolidinedions (aka glitazones) through mechanisms that are not fully known (Chih-Hao 2003; Yki-Jarvinen 2004). ).

Recently, drugs that act as ligands on the PPARγ receptor have emerged as therapeutic agents for type 2 diabetes (Yki-Jarvinen 2004). Compounds called thiazolidinediones or glitazones are drugs that alter the insulin resistance that is the pathophysiological basis of metabolic syndrome and type 2 diabetes development. Of these compounds, rosiglitazone and pioglitazone have been released as drugs, lowering fasting and postprandial glucose levels (which are indicated by pathological glucose tolerance tests), plasma insulin and free fatty acid concentrations. In this respect, the glitazone acts as insulin sensitizers.

However, these improvements involve weight gain and increase the amount of subcutaneous adipose tissue (Adams 1997). The use of thiazolidinediones is associated with weight gain as well as some patient groups causing peripheral edema, with fluid retention and plasma volume expansion. Weight gain and edema are associated with an increased risk of heart disease, which is why the US Food and Drug Administration (FDA) has included warnings in prescription information from rosiglitazone (available by Avandia) and pioglitazone (available by Takeda). Because of these side effects, especially patients with coronary heart disease refrain from using glitazone. There is certainly the possibility of developing a new drug that has weight loss activity, but does not have fluid retention, and which is compliant with insulin resistance. The effect of polyunsaturated fatty acids (PUFAs) on PPARs is not only due to fatty acid structure and affinity for receptors. Factors that contribute to the composition of intracellular non-esterified fatty acid (NEFA) concentrations are also important. The NEFA pool is influenced by the concentration of exogenous fatty acids entering the cell, the amount of endogenous synthetic fatty acids, the removal of NEFA through binding to fat or through oxidation pathways (Pawar 2003).

Although omega-3 fatty acids are weaker as agents of PPARs compared to pharmacological agents such as thioglitazone, these fatty acids have demonstrated improved glucose uptake and insulin sensitivity (Storlien 1987). When the ratio of polyunsaturated fatty acids to saturated fatty acids in food is increased, fat cells are known to be more insulin sensitive and to transport more glucose (Field 1990). Overall, these data show that 20- and 22-carbon fatty acids, ie EPA and DHA, may play a role in preventing insulin resistance.

PUFAs have not been widely used as pharmaceuticals due to their limited in vivo stability and lack of biological specificity. In order to alter or increase metabolic potency, many study groups have carried out studies on chemical modification of n-3 polysaturated fatty acids.

For example, the hypolipidemic effects of EPA are enhanced by introducing methyl or ethyl at the α- or β-position of EPA (Vaagenes 1999). EPA EE is ineffective, while the compounds reduce plasma free fatty acids.

In a recent study published by L. Larsen (Larsen 2005), the authors show that the α-methyl derivatives of EPA and DHA increased the activity of nuclear receptor PPARα and thus the expression of L-FABP compared to EPA / DHA. EPA having an ethyl group in the α-position activated PPARα while having the same strength as α-methyl EPA. The authors suggest that the delay in the degradation of α-methyl FA would be responsible for increasing their efficacy due to the reduction of β-oxidation in the mitochondria causing peroxysome oxidation.

α-methyl EPA is shown to be a more potent platelet aggregation inhibitor than EPA in both in vitro (Larsen 1998) and in vivo (Willumsen 1998).

Japanese Patent Application Laid-Open No. 05-00974 discloses DHA in which the alpha-position is substituted with an OH- group only as an intermediate. No investigation of the possibility of the pharmaceutical efficacy of this compound is disclosed.

 Also, Laxdale Limited has described the use of alpha-substituted derivatives of EPA in psychosis and central nervous system disease (US6689812).

                                             (A) α-methyl EPA

However, these modified fatty acids did not show satisfactory pharmaceutical activity and did not appear in the pharmaceutical industry market.

Summary of the Invention

It is an object of the present invention to provide new DHA-derivatives having therapeutic activity. Based on the invention, various aspects of the invention appear in the appended claims. Some of these aspects of the invention are as follows.

1.New compound, α-substituted polyunsaturated fatty acid derivatives

2. New Compounds for Use in Therapeutics and Therapies

3. Fatty Acid Composition or Pharmaceutical Composition Including New Compound

4. Fatty acid composition comprising a novel compound for use as a therapeutic or therapeutic

5. Use of new compounds for the manufacture of a medicament for preventing and / or treating diabetes in humans or animals

6. Use of new compounds for the manufacture of a medicament for treating and / or preventing obesity or overweight disease

7. Use of new compounds for the production of pharmaceuticals to control weight loss and / or prevent weight gain

8. New compounds for the production of pharmaceuticals for the treatment and / or prevention of amyloidos-related diseases

9. Use of new compounds for the production of pharmaceuticals for the treatment or prevention of a number of risk factors associated with cardiovascular diseases.

10. Use of new compounds for the production of pharmaceuticals for the prevention of strokes, cerebral ischemic attacks or cerebral or transient ischaemic attacks associated with atherosclerosis of several arteries.

11. Specific methods of treatment for diabetes diseases, preferably type 2 diabetes.

12. Methods of controlling weight loss, preventing weight gain and / or treating and / or preventing obesity or being overweight

13. Methods for treating and / or preventing diseases associated with amyloidosis

14. Methods for treating or preventing a plurality of risk factors associated with cardiovascular diseases.

15. How to prevent strokes, cerebral ischemic attacks or cerebral or transient ischaemic attacks associated with atherosclerosis of several arteries

16. Preparation of New Fatty Acid Analogues According to the Invention

The present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt, solvate, complex, or prodrug thereof:

 (I)

The R ₁ And R ₂ is the same or different and is a hydrogen atom, a hydroxy group, an alkyl group, a halogen atom, an alkoxy group, an acyloxy group, an acyl group, an alkenyl group, an alkynyl group, an aryl group, an alkylthio group, an alkoxycarbonyl group, an alkylsulfinyl group, alkyl A sulfonyl group, an amino group, and an alkylamino group;

X is a carboxylic acid group, a carboxylate group, or a carboxamide; And

The compound of formula (I) is (all-Z) -4,7,10,13,16,19-DHA (docosahexaenoic acid), alpha-methyl DHA, alpha-methyl DHA methyl ester, alpha-methyl DHA ethyl ester Or alpha-hydroxy DHA ethyl ester.

Wherein R ₁ is a hydrogen atom, R ₂ is not a hydrogen atom;

R ₂ is R ₁ is not a hydrogen atom if it is a hydrogen atom;

When R ₁ is a methyl group, R ₂ is not a hydrogen atom, and X is not a carboxylic acid group, methylcarboxylate or ethylcarboxylate;

When R ₂ is a methyl group, R ₁ is not a hydrogen atom, and X is not a carboxylic acid group, methylcarboxylate or ethylcarboxylate;

When R ₁ is a hydroxy group, R ₂ is a hydrogen atom and X is not ethylcarboxylate; And

When said R <_2> is a hydroxyl group, R <_1> is not a hydrogen atom and X is ethylcarboxylate.

In the compounds according to the invention, the alkyl group may be selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, n-hexyl, and benzyl; The halogen atom may be selected from the group consisting of fluorine, chlorine, bromine, and iodine; The alkoxy group may be selected from the group consisting of methoxy, ethoxy, propoxy, isopropoxy, sec-butoxy, phenoxy, benzyloxy, OCH ₂ CF ₃ and OCH ₂ CH ₂ OCH ₃ ; The acyloxy group may be selected from the group consisting of acetoxy, propionoxy, and propoxyoxy; The alkenyl group may be selected from the group consisting of allyl, 2-butenyl, and 3-hexenyl; The alkynyl group may be selected from the group consisting of propargyl, 2-butynyl, and 3-hexynyl; The aryl group is a phenyl group; The alkylthio group may be selected from the group consisting of methylthio, ethylthio, isopropylthio, and phenylthio; The alkoxycarbonyl group may be selected from the group consisting of methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl and butoxycarbonyl; The alkylsulfinyl group may be selected from the group consisting of methanesulfinyl, ethanesulfinyl and isopropanesulfinyl; The alkylsulfonyl group may be selected from the group consisting of methanesulfonyl, ethanesulfonyl and isopropanesulfonyl; The alkylamino group may be selected from the group consisting of methylamino, dimethylamino, ethylamino, and diethylamino; The carboxylate groups are ethyl carboxylate, methyl carboxylate, n-propyl carboxylate, isopropyl carboxylate, n-butyl carboxylate, sec-butyl carboxylate, and n-hexyl carboxylate May be selected from the group consisting of: The carboxamide group may be selected from the group consisting of primary carboxamide, N-methyl carboxamide, N, N-dimethyl carboxamide, N-ethyl carboxamide, and N, N-diethyl carboxamide have.

In one embodiment of the invention, R ₁ and R ₂ are selected from the group consisting of hydrogen atom, hydroxy group, alkyl group, halogen atom, alkoxy group, alkylthio group, alkylsulfinyl group, alkylsulfonyl group, amino group and alkylamino group.

In another embodiment of the invention, R ₁ and R ₂ are hydrogen atom, hydroxy group, C ₁ -C ₇ alkyl group, halogen atom, C ₁ -C ₇ alkoxy group, C ₁ -C ₇ alkylthio group, C ₁ -C ₇ alkylsulfinyl group, C ₁ -C ₇ alkylsulfonyl group, amino group, and C ₁ -C ₇ alkylamino group. And the C ₁ -C ₇ alkyl group can be methyl, ethyl or benzyl; The halogen atom may be fluorine or iodine; The C ₁ -C ₇ alkoxy group may be methoxy or ethoxy; The C ₁ -C ₇ alkylthio group can be methylthio, ethylthio, or phenylthio; The C ₁ -C ₇ alkylsulfinyl group may be ethanesulfinyl; The C ₁ -C ₇ alkylsulfonyl group may be ethanesulfonyl; The C ₁ -C ₇ alkylamino group can be ethylamino or diethylamino; And X may be an ethylcarboxylate or carboxyamide group.

In another embodiment of the present invention, R ₁ and R ₂ are a hydrogen atom, a C ₂ -C ₇ alkyl group, a halogen atom, a C ₁ -C ₇ alkoxy group, a C ₁ -C ₇ alkylthio group, C ₁ -C ₇ An alkylsulfinyl group, a C ₁ -C ₇ alkylsulfonyl group, an amino group, and a C ₁ -C ₇ alkylamino group; X is a carboxylate. And the C ₂ -C ₇ alkyl group can be methyl, ethyl or benzyl; The halogen atom may be fluorine or iodine; The C ₁ -C ₇ alkoxy group may be methoxy or ethoxy; The C ₁ -C ₇ alkylthio group can be methylthio, ethylthio, or phenylthio; The C ₁ -C ₇ alkylsulfinyl group may be ethanesulfinyl; The C ₁ -C ₇ alkylsulfonyl group may be ethanesulfonyl; The C ₁ -C ₇ alkylamino group can be ethylamino or diethylamino; And X is ethylcarboxylate.

In the compounds according to formula (I) of the invention, R ₁ And R ₂ may be the same or different from one another. If R ₁ and R ₂ are different from each other, the compound of formula (I) may exist in stereoisomeric form. It will be understood that the present invention includes all optical isomers of component (I) and mixtures thereof including racemic compounds.

Thus, the present invention, R ₁ And compounds of formula (I) which are optically pure as racemic or (S) or (R) enantiomers, wherein R ₂ is different. Thus, the present invention, R ₁ And compounds of formula (I) which are optically pure as racemic or (S) or (R) stereoisomers, wherein R ₂ is different.

As defined above, there are enantiomers of the above compound of formula (I) within the scope of the present invention. In addition, the enantiomers of the DHA derivatives according to the invention may be in carboxylic acid form or pharmaceutically acceptable salts, esters, anhydrides, or amides (primary, secondary, tertiary). The acid derivative may be in phospholipid or tri-, di- or monoglyceride form.

In one embodiment of the compound of formula (I) according to the invention, R ₁ And one of R ₂ is a C ₂ -C ₇ alkyl group, ie ethyl or benzyl, and the other is a hydrogen atom. Of these, the alkyl group is preferably ethyl.

In another embodiment of the compound of formula (I) according to the invention, R ₁ And one of R ₂ is an alkoxy group, ie ethoxy or methoxy, and the other is a hydrogen atom.

In another embodiment of the compound of formula (I) according to the invention, R ₁ And one of R ₂ is a halogen atom, ie fluorine or iodine, and the other is a hydrogen atom.

In another embodiment of the compound of formula (I) according to the invention, R ₁ And one of R ₂ is an alkylthio group, ie ethylthio, methylthio or phenylthio, and the other is a hydrogen atom. Of these, alkylthio groups are preferably ethylthio.

In another embodiment of the compound of formula (I) according to the invention, R ₁ And one of R ₂ is an alkylsulfonyl group, ie ethylsulfonyl, and the other is a hydrogen atom.

In another embodiment of the compound of formula (I) according to the invention, R ₁ And one of R ₂ is an amino group, and the other is a hydrogen atom.

In another embodiment of the compound of formula (I) according to the invention, R ₁ And one of R ₂ is an alkylamino group, ie ethylamino or diethylamino, and the other is a hydrogen atom.

In another embodiment of the compound of formula (I) according to the invention, R ₁ And R ₂ are the same and are a C ₁ -C ₇ alkyl group, preferably a methyl group or an ethyl group.

In a preferred embodiment of the compound of formula (I), X is a carboxylate, ie ethyl carboxylate.

The compounds according to the invention may exist in the form of phospholipids, tri-, di-, monoglycerides or in the form of free acids.

The alpha-substituted DHA derivatives according to the present invention show surprising results on pharmaceutical activity. In particular, the fatty acid derivatives according to the invention have enormous potential for use in the treatment and / or prevention of diabetes and its predecessors.

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

The present invention relates to the preparation of compounds of formula (I). For example, the compound of formula (I) may be prepared from (all-Z) -4,7,10,13,16,19-DHA (docosahexaenoic acid). It can be obtained from an animal source such as DHA, vegetables, microbial and / or marine fish oil. Another important advantage of compounds of formula (I) is that fatty acid analogs can be prepared directly from (all-Z) -4,7,10,13,16,19-DHA (docosahexaenoic acid).

In a preferred embodiment of the invention, the fatty acid analog of formula (I) is prepared from DHA, which DHA is obtained from at least one of at least one vegetable, microorganism or animal and combinations thereof. The present invention therefore encompasses derivatives prepared from oils comprising DHA obtained from microorganisms. The DHA is preferably produced from marine oil such as fish oil.

Another aspect of the invention relates to a pharmaceutical composition comprising a compound of formula (I) as an active ingredient. The pharmaceutical composition may further comprise a pharmaceutically acceptable carrier. The pharmaceutical compositions according to the invention are formulated for oral administration, ie in capsule or sachet form. The daily dose of the compound of formula (I) according to the invention is 10 mg to 10 g, in particular 100 mg to 1 g.

In addition, the present invention relates to fatty acid compositions comprising a compound of formula (I). At least 60% or at least 90% by weight of the fatty acid composition is composed of the compound. The fatty acid composition is (all-Z) -5,8, ll, 14,17-eicosapentaenoic acid (EPA), (all-Z) -4,7,10,13,16,19-docosahexaenoic acid (DHA), (all-Z) -6,9,12,15,18-HPA (heneicosapentaenoic acid), and / or (all-Z) -7,10,13,16,19-DPA (docosapentaenoic acid) . The fatty acid may be present in the form of derivatives. The fatty acid composition according to the present invention may further comprise a pharmaceutically acceptable antioxidant such as tocopherol. The scope of the present invention also includes the use of the above-mentioned fatty acid composition as a medicament.

In another aspect of the present invention, the present invention provides a pharmaceutical composition for controlling weight loss and / or preventing weight gain; And / or the manufacture of a medicament for the treatment and / or prevention of obesity or overweight disease; Preparation of a medicament for the prevention and / or treatment of diabetes in animals, in particular type 2 diabetes; Preparation of a medicament for the treatment and / or prevention of amyloidosis-associated diseases; Treatment or prevention of a number of risk factors associated with cardiovascular diseases, preferably treatment of elevated blood lipids, stroke associated with atherosclerosis of several arteries, cerebral ischemic attack or transient The use of a compound of formula (I) for the manufacture of a medicament for the prevention of cerebral or transient ischaemic attacks.

In addition, the present invention provides a weight loss control method and / or weight gain prevention method; And / or methods of treating and / or preventing obesity or being overweight; Methods of preventing and / or treating diabetes in animals, especially type 2 diabetes; Methods of treating and / or preventing amyloidosis associated diseases; Methods of treating or preventing a number of risk factors associated with cardiovascular diseases; As a method of preventing stroke, cerebral ischemic attack or transient ischemic attacks associated with atherosclerosis of several arteries, a pharmaceutically effective amount of the compound of formula (I) A method of administering to a human.

Brief description of the drawings

1 shows a schematic overview of the free fatty acid pool theory.

Figure 2 shows an overview of the methods and models used in the present invention to demonstrate effects on metabolic syndrome and type 2 diabetes.

Figure 3 shows the free fatty acid concentrations of the different compounds according to the invention in the adipose tissue of animals receiving different compounds according to the invention at a concentration of 1.5% of the total fat concentration.

Figure 4 shows the intracellular DHA concentration in the adipose tissue of animals receiving different compounds according to the invention at a concentration of 1.5% of the total fat concentration.

Figure 5 shows the binding affinity for PPAR receptors of different compounds according to the invention.

Figure 6 shows the binding affinity for nuclear receptor PPARα of different compounds according to the present invention.

Figure 7 shows the binding affinity for the nuclear receptor RXRα of different compounds according to the invention.

Figure 8 shows luciferase release from infected cells treated with different compounds according to the present invention.

9 shows a study design of a four block experiment.

FIG. 10 shows body weight change during 2 weeks diet control after 8 weeks HF diet.

Figure 11 shows the results of luciferase activity, ie endogenous PPARγ activity.

Figure 12 shows endogenous luciferase activity in different compounds according to the invention compared to DHA.

FIG. 13 shows a typical blood glucose output curve before and after administration of a compound having an insulin resistance reducing effect to an insulin resistant animal.

14, 15 and 16 show the different effects on the metabolic syndrome and insulin resistance of the DHA derivative according to the present invention.

DETAILED DESCRIPTION OF THE INVENTION

In a recent study according to the invention, new DHA derivatives have been prepared and have shown excellent pharmaceutical activity.

Fatty acids enter the cell passively or through G-protein coupled transporter systems such as fatty acid transport proteins. They are temporarily bound in cells by binding proteins (Fatty acid binding proteins, FABPs), which are important in directing fatty acids to various intracellular compartments for metabolism and gene expression. Play a role (Pawar & Jump 2003). (Figure 1. Hepatocytes).

The esterification of fatty acids with triglycerides, polar lipids and cholesterol esters and their beta-oxidation (mitochondria and peroxysomes) requires the conversion of fatty acids to acyl CoA. Another route uses nonesterified fatty acids as substrates, such as microsomal NADPH-dependent mono-oxidation and eicosanoid synthesis. All these reactions are likely to affect the cell concentration of free fatty acids (deesterifications) that can be used as ligands for nuclear receptors and thereby the amount and type of fatty acids. Since PPARs are known to bind to esters and fatty acids, it can be predicted that free fatty acid pool compositions are important determinants of PPAR activity regulation.

The composition of the free fatty acid pool is influenced by the concentration of exogenous fatty acids entering the cell and their removal rate through the aforementioned pathways. Short or medium chain fatty acids are efficiently added to these pathways, and in fact only long chain polysaturated fatty acids can be liganded to nuclear receptors. In addition, fatty acid structures can also be important determinants. Although a series of mono and polysaturated fatty acids showed affinity for PPARα receptors, EPA and DHA showed the highest binding capacity in experiments with rat hepatocytes (Pawar & Jump 2003).

To find fatty acid candidates that can be genetically modified by interaction with nuclear receptors such as PPARs, it is important to demonstrate that individual fatty acids will be enriched in the free fatty acid pool.

DHA entering the cell is rapidly converted into fatty acyl-CoA thioesters and binds to phospholipids. And because of this, the intracellular DHA concentration is relatively low. In addition, these DHA-CoA is a substrate mainly for β-oxidation in peroxysomes that allow for retroconvertion of DHA to EPA (see FIG. 1). Due to its rapid binding to triglycerides and the oxidation pathway, DHA will not stay long in the free fatty acid pool. For this reason, the effect of DHA on gene expression is probably limited.

The present invention is intended to accumulate fatty acid derivatives in the free fatty acid pool rather than to bind to phospholipids. The inventors have surprisingly found that the introduction of at least one substituent at the DHA α-position will lower the oxidation rate and lower the binding rate to triglycerides. This will increase gene expression effects, especially since DHA derivatives will accumulate in tissues such as liver, muscle, and adipocytes, and will stimulate local nuclear receptor activity even greater than DHA.

Other substituents according to the present invention will exhibit varying affinity of derivatives for fatty acid binding receptors. In addition, affinity changes to fatty acid binding proteins have the potential to alter the biological activity of the α-substituted DHA derivatives of these formula (I). Overall, these changes can increase the therapeutic effect of the DHA derivatives according to the invention compared to DHA.

EPA ((all-Z) -5,8, l l, 14,17-eicosapentaenoic acid) is initially alkylated at the α- and β-positions to inhibit mitochondrial β-oxidation. DHA is not oxidized in the mitochondria but instead binds to phospholipids. Nevertheless, some DHA in peroxysomes is reconverted to EPA. Substituents at the α-position of EPA and DHA occur because of affecting different metabolic pathways. It is already known that α-methyl EPA and β-methyl EPA bind to phospholipids and triglycerides while α-ethyl EPA is not bound (Larsen 1998). In this study, the derivatives were tested as substrates and / or inhibitors of enzymes involved in the eicosanoid cascade. Since most substrates for these enzymes are fatty acids free from phospholipids, the derivatives are preferably bound to phospholipids. In contrast, as mentioned above, we want derivatives that do not bind to lipids and accumulate in the NEFA pool.

Throughout this specification, the abbreviation “PRB-x”, where x is an integer, will be used when describing certain compounds according to the invention. Structural formulas and common names of these compounds are as follows.

PRB-1 corresponds to the compound of formula (I) wherein one of R ₁ and R ₂ is methyl and the other is hydrogen and X is ethyl carboxylate.

PRB-2 corresponds to the compound of formula (I) wherein one of R ₁ and R ₂ is ethyl and the other is hydrogen and X is ethyl carboxylate.

PRB-3 corresponds to the compound of formula (I) wherein one of R ₁ and R ₂ is ethoxy and the other is hydrogen and X is ethyl carboxylate.

PRB-4 corresponds to the compound of formula (I) wherein one of R ₁ and R ₂ is fluorine and the other is hydrogen and X is ethyl carboxylate.

PRB-5 corresponds to the compound of formula (I) wherein R ₁ and R ₂ are methyl and X is ethyl carboxylate.

PRB-6 corresponds to the compound of formula (I) wherein R ₁ or R ₂ is methylthio and X is ethyl carboxylate.

PRB-7 corresponds to the compound of formula (I), wherein one of R ₁ and R ₂ is ethylthio, the other is hydrogen, and X is ethyl carboxylate.

PRB-8 corresponds to the compound of formula (I) wherein R ₁ and R ₂ are ethyl and X is ethyl carboxylate.

PRB-9 corresponds to the compound of formula (I), wherein one of R ₁ and R ₂ is benzyl, the other is hydrogen, and X is ethyl carboxylate.

PRB-10 corresponds to the compound of formula (I) wherein one of R ₁ and R ₂ is ethylsulfonyl, the other is hydrogen, and X is ethyl carboxylate.

PRB-11 corresponds to a compound of formula (I) wherein one of R ₁ and R ₂ is phenylthio, the other is hydrogen, and X is ethyl carboxylate.

PRB-12 corresponds to the compound of formula (I) wherein one of R ₁ and R ₂ is hydroxy, the other is hydrogen and X is ethyl carboxylate.

PRB-13 corresponds to a compound of formula (I) wherein one of R ₁ and R ₂ is methyl, the other is hydrogen, and X is primary carboxamide.

PRB-14 corresponds to the compound of formula (I), wherein one of R ₁ and R ₂ is methoxy, the other is hydrogen, and X is ethyl carboxylate.

PRB-15 corresponds to the compound of formula (I), wherein one of R ₁ and R ₂ is iodine, the other is hydrogen, and X is ethyl carboxylate.

PRB-17 corresponds to a compound of formula (I) wherein one of R ₁ and R ₂ is amino, the other is hydrogen, and X is ethyl carboxylate.

PRB-20 corresponds to the (S) stereoisomer of Formula (I), wherein one of R ₁ and R ₂ is ethyl, the other is hydrogen, and X is ethyl carboxylate.

PRB-23 corresponds to the (R) stereoisomer of formula (I), wherein one of R ₁ and R ₂ is ethyl, the other is hydrogen, and X is ethyl carboxylate.

PRB-24 corresponds to the (R) stereoisomer of formula (I), wherein one of R ₁ and R ₂ is N-phthalimide, the other is hydrogen, and X is ethyl carboxylate.

PRB-25 corresponds to the compound of formula (I), wherein one of R ₁ and R ₂ is ethyl-amino, the other is hydrogen, and X is primary carboxamide.

PRB-26 corresponds to a compound of formula (I) wherein one of R ₁ and R ₂ is diethyl-amino, the other is hydrogen, and X is ethyl carboxylate.

The most preferred compound according to the invention is PRB-2. Other preferred compounds according to the invention are PRB-5, PRB-7, and PRB-8.

It is to be understood that the present invention includes all possible pharmaceutically acceptable salts, solvates, complexes or prodrugs of the compounds of formula (I).

A “prodrug” may or may not have its own pharmacological activity, may be administered by oral or parenteral methods, and then bioactivated (eg, metabolized) in the body. It can be a drug of the present invention having a pharmaceutical activity.

If X is a carboxylic acid, the present invention also includes salts of carboxylic acids. Preferably the pharmaceutically acceptable salts of the carboxyl groups include metal salts such as aluminum, alkali metal salts such as lithium, sodium, or potassium, alkaline metal salts such as calcium or magnesium and ammonium or chelated ammonium salts.

“Amount effective for treatment” refers to the amount of therapeutic agent effective to achieve the desired purpose. Determination of the appropriate range of effective amounts of each nitric oxide adduct is within the range well known in the art, as it varies with the individual patient. In general, the dosage regimen for treating a disease using the compounds and / or compositions of the present invention is selected by a variety of factors including the patient's body type, age, weight, sex, diet and medical condition.

"Medicament" means a compound according to formula (I) in any form that can be used for medical purposes, and exists in various forms such as, for example, a medicine, drug preparation or product, food, food or food supplement .

The term "therapy" in the context of the present invention includes "prophylaxis" unless there is a specific indication to the contrary. The terms "therapeutic" and "therapeutically" should be constructed accordingly.

Treatment (or treatment) includes all therapeutic activities beneficial to humans or non-human animals. In particular, treatment with mammals is preferred. Both treatments for humans and animals are within the scope of the present invention. The treatment may be for an existing disease or may be prophylactic. It may be treatment for an adult, adolescent, infant, fetus or a portion of the aforementioned (ie organ, tissue, cell or nucleic acid molecule). "Chronic treatment" means treatment that lasts for weeks or years.

A "therapeutically or pharmaceutically active amount" refers to an amount that exhibits the desired pharmacological and / or therapeutic effect. The compounds according to the invention can be included, for example, in food, food supplements, nutritional supplements, or foods.

Alpha-substituted DHA derivatives and EPA (or DHA for this) are esters of alpha-derivatives, EPA, and a mixture of glycerol catalyzed by Novozym 435 (commercial lipase made from fixed form Candida antarctica) It may be combined and mixed with each other in triglyceride form by means of oxidization.

Compounds of formula (I) have activity as drugs, in particular as triggers of nuclear receptor activity. Thus, the present invention also relates to the drugs of formula (I), pharmaceutically acceptable salts, solvates, complexes or prodrugs thereof, for use as therapeutics and / or therapeutic agents, as mentioned above. Preferably, the new compounds of the invention, their pharmaceutically acceptable salts, solvates, complexes or prodrugs may be used as follows:

For preventing and / or treating diabetes in humans or animals;

For controlling weight loss and / or for preventing weight gain;

For preventing and / or treating obesity or overweight disease in humans or animals;

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

For the treatment or prevention of a number of risk factors associated with cardiovascular diseases;

For the prevention of strokes, cerebral ischemic attacks or cerebral or transient ischaemic attacks associated with atherosclerosis of several arteries;

For the treatment of TBC or HIV.

There are two types of diabetes. One is insulin-dependent diabetes mellitus (IDDM) as type 1 diabetes and the other is non-insulin-dependent diabetes mellitus (NIDDM) as type 2 diabetes. Type 2 diabetes is primarily associated with obesity / overweight and lack of exercise, which are chronic in adults, and are manifested by weakened insulin sensitivity called peripheral insulin resistance. This compensatively increases insulin production. This stage, which occurs before the complete recovery of type 2 diabetes, is called metabolic syndrome, and hyperinsulinemia, insulin resistance, obesity, glucose tolerance, high blood pressure, abnormal blood lipids, hypercoagulopathia, lipid abnormalities and inflammation Characteristic and often causes atherosclerosis of the aorta. Subsequently, when insulin production is detected, type 2 diabetes occurs.

In a preferred embodiment of the invention, the compounds according to formula (I) can be used to treat type 2 diabetes. Compounds according to formula (I) may also be used for secondary diabetes, lipoatrophic, muscle relaxation, such as metabolic syndrome, pancreatic, extrapancreatic / endocrine or drug-induced diabetes. It can be used to treat other types of diabetes, selected from the group consisting of exceptional forms of diabetes, such as diseases caused by myatonic or insulin receptor disorders. The invention also includes the treatment of type 2 diabetes. Preferably, the compounds of formula (I) as defined above can activate nuclear receptors, in particular peroxisome proliferator-activated receptor (PPAR) α and / or γ.

Compounds of formula (I) can be used to treat and / or prevent obesity. Obesity is always associated with increased insulin resistance, and obese people are at higher risk for type 2 diabetes, a major factor in the development of heart disease. In Western societies, the obese population is on the rise and is associated with social problems as well as many problems such as reduced lifespan and diabetes, insulin resistance and hypertension. The present invention frees long-term cravings for drugs that will reduce the amount or weight of adipose tissue so that they usually have normal weight without serious side effects in people who are obese.

The compounds according to formula (I) can be used for the prophylaxis and / or treatment of diseases associated with amyloidosis. Diseases associated with amyloidosis and / or diseases associated with amylose accumulation, especially as a result of fibril or plaque formation, Alzheimer's disease, dementia, Parkinson's disease, amyotropic lateral sclerosis, infectious Spongiform encephalopathies Creutzfeld- jacob disease, cystic fibrosis, primary or secondary renal amyloidoses, IgA nephropathy, arteries, myocardium ) And amyloid accumulation in neutral tissues. These diseases are sporadic, hereditary and are also associated with infections such as TBC or HIV. And even if heredity appears earlier, it often appears later in life. Each disease is associated with a specific protein or aggregation of these proteins is believed to be a direct source of the pathological disease associated with the disease. Treatment of diseases associated with amyloidosis can occur either acutely or chronically.

In addition, the compounds of formula (I) are useful for the treatment of reduced amyloid aggregates, for the prevention of misfolding of proteins that may form fibrils or plaques, for the formation of precursor proteins such as Aβ-proteins (amyloid beta proteins). It can be used for treatment due to the reduction of and for prophylaxis and / or treatment due to inhibiting or lowering protein fibrils, aggregates, or plaque formation. Fibrillar accumulation or formation prevention by administration of a compound of formula (I), as described above, is also within the scope of the present invention. In one embodiment of the present invention, as mentioned above, novel compounds, pharmaceutically acceptable salts, solvates, complexes or prodrugs thereof are used for the treatment of TBC (tuberculosis) or HIV (human immunodeficiency virus).

In addition, the compound of formula (I) is administered to patients suffering from atherosclerosis of the cerebral arteries, for example stroke, transient ischemic attacks, thereby reducing the risk of more serious development, which may be fatal.

The compounds of formula (I) can also be used to treat elevated blood lipids in humans.

In addition, as described above, the compounds of formula (I) are useful for preventing and treating a number of risk factors for cardiovascular diseases such as high blood pressure, hypertriglyceridemia and high coagulation factor VII of phospholipid complex activity. . Preferably the compound of formula (I) of the present invention can be used to treat elevated blood lipids in humans.

The compounds of formula (I) and their pharmaceutically acceptable salts, solvates, prodrugs or complexes thereof may be used by themselves, but generally the compounds of formula (I) (active ingredient) are pharmaceutically acceptable It is administered in the form of a pharmaceutical composition in combination with an adjuvant, diluent or carrier.

The present invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I) and a pharmaceutically acceptable carrier, diluent or additive, including combinations thereof.

It is a composition comprising a therapeutically effective amount of a pharmaceutically active ingredient. It is preferred to include a pharmaceutically acceptable carrier, diluent, or additive, including combinations thereof. Acceptable carriers or diluents for such treatments are those well known in the art. The choice of pharmaceutical carrier, excipient or diluent may be selected according to the desired route of administration and standard practice of the pharmaceutical field. The pharmaceutical composition may further include a carrier, an additive, or a diluent, a suitable binder, a lubricant, a suspending agent, a coating agent, a dissolving agent alone or a mixture thereof.

The pharmaceutical compositions of the present invention include preservatives, solubilizers, stabilizers, wetting agents, emulsifiers, sweeteners, colorants, flavors, olfactory stimulants, salt compounds of the invention (will be provided in pharmaceutically acceptable salt form), buffers, coatings, One or more antioxidants, suspending agents, adjuvants, additives and diluents.

The pharmaceutical composition of the present invention is formulated for oral administration to humans or animals. The pharmaceutical compositions may also be formulated for administration via other routes through which the active ingredient may be effectively absorbed and used, i.e., via the intravenous, subcutaneous, intramuscular, nasal, rectal, vaginal or topical routes.

In one embodiment of the invention, the pharmaceutical composition is in the form of a capsule, it may be a microcapsules to make powder or dead body. The capsule may be scented. Also included in this embodiment is a capsule wherein both the capsule and the encapsulated fatty acid composition according to the invention are fragrant. Users are attracted to the scented capsules. For the aforementioned therapeutic uses, the dosage will naturally vary depending on the compound used, the dosage form, the purpose of treatment, and the manifestation of the disease present.

The pharmaceutical composition may be formulated to have a daily dosage of 10 mg to 10 g. Preferably, the daily dosage of the pharmaceutical composition is formulated to be between 50 mg and 5 g. Most preferably, the daily dosage of the pharmaceutical composition is prepared to be from 100 mg to 1 g. Refers to the daily dose of 24 hours. The dosage will naturally vary depending on the compound used, the dosage form, the purpose of treatment, and the indication of the disease present. In general, the physician will determine the actual dosage amount that is most appropriate for the individual patient. The number of doses under a particular dosage for a patient can vary depending on the activity of the compound used, the metabolic stability and duration of the compound, the age, weight, health condition, sex, diet, mode and time of administration, rate of release, drug combination, It depends on various factors, including the severity of the particular condition and the patient being treated. The pharmaceutical and / or pharmaceutical compositions of the invention may be administered according to a regimen of 1 to 10, such as once or twice daily. For oral or parenteral administration to human patients, the medicament may be taken at one time or in divided doses.

Another aspect of the invention relates to a fatty acid composition comprising the compound of formula (I) above. Fatty acid compositions comprising a compound of formula (I) increase the natural biological effect of DHA, which is the result of gene expression, and its derivatives according to the invention will accumulate in the free fatty acid pool.

The fatty acid composition may comprise from 60 to 100% by weight of the compound of formula (I), all weight percentages being relative to the total weight of the fatty acid composition. In a preferred embodiment of the invention, at least 80% by weight of the fatty acid composition consists of the compound of formula (I). More preferably, the compound of formula (I) is included in at least 90% by weight of the fatty acid composition. Most preferably, the compound of formula (I) is included in at least 95% by weight of the fatty acid composition.

The fatty acid composition is (all-Z) -5,8, ll, 14,17-eicosapentaenoic acid (EPA), (all-Z) -4,7,10,13,16,19 Docosahexaenoic acid (DHA), (all-Z) -6,9, 12,15, 18-heneicosapentaenoic acid (HPA), and (all-Z) -7,10,13,16,19-docosapentaenoic acid (DPAn-3), (all-Z) -8, l1,14,17-eicosatetraenoic acid (eicosatetraenoic acid, ETAn-3), or a mixture thereof. The fatty acid composition may also comprise (all-Z) -4,7,10,13,16-docosapentaenoic acid (DPAn-6) and / or (all-Z) -5,8, l1,14 -Eicosatetraenoic acid (ARA), or derivatives thereof. In addition, the fatty acid composition may further comprise at least these fatty acid compositions or mixtures thereof in the form of derivatives. As mentioned above, the derivatives are suitably substituted in the same manner as the DHA derivatives of the formula (I).

The fatty acid composition according to the present invention comprises at least 1% by weight or 1 (all-Z omega-3) -6,9,12,15, 18-heneicosapentaenoic acid (HPA), or derivatives thereof. In an amount of 1% by weight.

In addition, the fatty acid composition according to the present invention may include at least 1.5% by weight or at least 3% by weight of omega-3 fatty acids or derivatives thereof having 20, 21, or 22 carbon atoms other than EPA and DHA.

In an embodiment of the invention, the fatty acid composition is a pharmaceutical composition, nutritional composition or diet composition. The fatty acid composition may further comprise an effective amount of a pharmaceutically acceptable antioxidant. In a preferred embodiment, the fatty acid composition further comprises tocopherol, or a tocopherol mixture, up to 4 mg per g of the total weight of the fatty acid composition. Preferably, the fatty acid composition comprises 0.2 to 0.4 mg per gram of tocopherol relative to the total weight of the composition.

In another aspect of the invention, the invention provides a therapeutic agent and / or treatment for a fatty acid composition comprising a compound of formula (I) or a pharmaceutically acceptable salt, solvate, precursor, or complex thereof, as mentioned above. Provide for the purpose. Such fatty acid compositions can be used for the prophylaxis and / or treatment of diseases for using the compounds of formula (I) already mentioned.

When the fatty acid composition is used as a medicament, a therapeutic or pharmaceutically active amount will be administered.

In a preferred embodiment, the fatty acid composition is administered orally to humans or animals.

The present invention, as mentioned above, provides the following uses of a compound of formula (I), a pharmaceutically acceptable salt, solvate, precursor or complex thereof. The present invention provides a pharmaceutical preparation for weight loss control and / or weight gain prevention; And / or the manufacture of a medicament for the treatment and / or prevention of obesity or overweight disease; Preparation of a medicament for preventing and / or treating diabetes in humans or animals; Preparation of a medicament for preventing and treating a number of risk factors for cardiovascular diseases such as hypertension, hypertriglyceridemia and high coagulation factor VII of phospholipid complex activity; Preparation of a medicament for treating TBC or HIV; Preparation of a medicament for preventing stroke, cerebral ischemic attack or transient ischemic attack associated with atherosclerosis of a plurality of arteries; Preparation of a medicament for lowering triglycerides in mammalian blood and / or a medicament for raising HDL cholesterol levels in serum of human patients and / or for the preparation and treatment of a medicament for the treatment and / or prophylaxis of multiple metabolic syndrome called "metabolism syndrome". Provides a use for The invention also relates to the use of alpha-hydroxy-DHA for the preparation of the abovementioned medicaments.

The present invention further relates to a method for controlling weight loss and a method for preventing weight gain, wherein a fatty acid composition comprising at least one compound of formula (I) mentioned above is administered to a human or animal.

The invention also relates to a method for the treatment and / or prevention of obesity and overweight, wherein a fatty acid composition comprising at least one compound of formula (I) mentioned above is administered to a human or animal.

Furthermore, the present invention relates to a method for the prevention and / or treatment of diabetes, wherein a fatty acid composition comprising at least one compound of formula (I) mentioned above is administered to a human or animal. In particular, diabetes is type 2 diabetes.

Another aspect of the invention relates to the following method wherein a fatty acid composition comprising at least one compound of the formula (I) mentioned above is administered to a human or animal:

-Methods of treating and / or preventing diseases associated with amyloidosis;

How to treat or prevent a number of risk factors associated with cardiovascular diseases;

-Methods of preventing stroke, cerebral ischemic attacks or cerebral or transient ischaemic attacks associated with atherosclerosis of several arteries.

Fatty acid derivatives of formula (I) may be said to be most effective prepared using DHA. If the starting material is not pure DHA (ie not 100% DHA), the fatty acid composition will comprise some amount of the aforementioned DHA derivative, a fatty acid composition other than DHA, which fatty acid compositions of formula (I) Substituted in the same way as novel fatty acid analogs. Such embodiments are also included herein.

In another embodiment of the present invention, the compound of formula (I) is prepared using (all-Z) -4,7, 10,13,16,19-docosahexaenoic acid (DHA), wherein DHA is obtained from vegetables, microorganisms and / or animals or mixtures thereof. The DHA is most preferably obtained from marine oil such as fish oil.

The fatty acids of the composition may also be obtained from vegetables, microorganisms or animals or mixtures thereof. Thus, the present invention also encompasses fatty acid compositions prepared with microbial oils.

The present invention provides a process for preparing novel fatty acid analogs of the compounds of formula (I), as mentioned above.

DHA is obtained from biological sources such as marine, microbial, or vegetable fats. All possible raw materials are fatty acid mixtures in the form of triglycerides in which DHA forms part of the fatty acid. Typical DHA concentrations are 40% in microbial fats and 10-25% in marine oils. Vegetable fats containing DHA are under development, and fats with high levels of DHA are expected in the future.

The first step will always be the conversion of triglycerides to free fatty acids or monoesters. Preferred esters are methyl or ethyl esters, but other esters are possible. In this process, the fatty acids bound together in three to three triglycerides are separated from each other and thereby become separable. Various methods of separating DHA from other fatty acids are available. Short path distillation, which separates fatty acids by volatility, and urea precipitation, which separates fatty acids by unsaturation, are most common. . Other known methods include a silver nitrate complex formation method for separating fatty acids according to the degree of unsaturation, a short pass distillation method and a countercurrent extraction method using supercritical carbon dioxide, and an esterification reaction catalyzed by fatty acid selective lipase.

The most important thing for producing pure DHA is to separate DHA from other C20-22 highly unsaturated fatty acids present in all possible ingredients. These fatty acids have properties similar to DHA and are not separated to a sufficient degree by the methods mentioned above. Some microorganisms' high DHA fats, which have very low concentrations of C20-22 highly unsaturated fatty acids, may have a purity of 90% or higher when separated by short pass distillation or by mixing with other methods mentioned above.

Most DHA containing fats also contain significant amounts of C20-22 highly unsaturated fatty acids such as EPA (20: 5n-3), n-3DPA (22: 5n-3), HPA (21: 5n-3), and Other materials. The only way to separate DHA from these fatty acids is preparative High Performance Liquid Chromatography, where the stationary phase is silica gel or silica gel impregnated with silver nitrate and the mobile phase is an optional organic solvent or supercritical carbon dioxide. Using this method, DHA with a purity of 97% or higher can be obtained. However, as the DHA concentration increases, the production cost increases, such as the 97% DHA production cost is more than five times the 90% DHA production cost.

DHA with a purity of 90, 95 or 97% contains a small amount of other fatty acids. For example, 97% pure DHA contains n-3DPA (22: 5n-3) and also contains long chain fatty acids such as EPA (20: 5n-3), HPA (21: 5n-3) and other substances. Include. However, other fatty acids will react in a similar manner to DHA to provide alpha-substituted derivatives.

Because only DHA and n-6DPA (and 22: 5n-6 present at very low concentrations) are known as fatty acids capable of providing gamma-lactones by ring bonds of the first double bond, organic synthesis Purification methods will be provided. Lactonisation and hydrolysis following purification may be possible, reverting to DHA, but this will be much more expensive than HPLC.

In one embodiment of the invention, the compound of formula (I), wherein R ₁ (or R ₂ ) is hydrogen, is prepared by the following procedure (Scheme 1). These procedures can be suitably modified and used for the preparation of compounds of formula (I) wherein R ₁ and R ₂ are C ₁ -C ₇ alkyl groups, benzyl, halogen, benzyl, alkenyl, or alkynyl.

Compounds of formula (I) wherein R ₁ is hydrogen and R ₂ is a C ₁ -C ₇ alkyl group, benzyl, halogen, benzyl, alkenyl, or alkynyl, may be selected from tetrahydrofuran, di Ester enolates are provided by reacting DHA esters with lithium diisopropylamine or strong non-nucleophilic bases such as potassium / sodium hexamethyldisilazideane in a solvent such as ethyl ester. (Course 1).

These ester enolates are ethyliodine, halogenated alkyls such as benzyl chloride, acetyl chlorides, halogenated acyls such as benzoylbromide, carboxylic anhydrides such as acetic anhydride, or N-fluorobenzene sulfonamide (NFSI) and the like. Reaction with an electrophilic reagent, such as an electrophilic halogenation reagent, provides a monosubstituted derivative (process 2). The ester is further hydrolyzed to carboxylic acid derivative in ethanol or methanol solvent by addition of a base such as lithium / sodium hydroxide in aqueous solution, between 15-40 ° C.

Claisen condensation of DHA EE occurs during the treatment of DHA EE with strong bases. This condensation product will have an interesting biological activity. Thus, in one embodiment of the invention, the above-mentioned condensation (intermediate) products and the use of these products for the treatment and / or prevention of the diseases according to the invention are disclosed.

In another embodiment of the present invention, the compound of formula (I) is synthesized through the following procedure (Scheme 2).

Compounds of formula (I) wherein R ₁ is hydrogen and R ₂ is a hydroxy, alkoxy group, acylcock group can be reacted with a DHA ester in a solvent such as tetrahydrofuran, diethyl ester at a temperature of -60 to -78 ° C. Ester enolates are provided by reacting with a strong non-nucleophilic base such as lithium diisopropylamine, or potassium / sodium hexamethyldisilazideane (process 4). This ester enolate is combined with other sources such as dimethyldioxirane, 2- (phenylsulfonyl) -3-phenyloxaziridine, dimethyldioxirane and other additives such as oxygen molecules and trimethylphosphite or Ni (II) complexes. Reaction with other catalysts such as to provide the alpha-hydroxy DHA ester (process 5). The reaction of a base such as sodium hydride with a secondary alcohol in THF or DMF solvent produces an alkoxide, which is an alkyl iodide such as methyl iodide, ethyl iodide, benzyl bromide React with different electrophilic reagents such as alkyliodide or acyl halides such as acetyl chloride, benzoyl bromide (process 6). The ester is hydrolyzed to carboxylic acid in ethanol or methanol solvent by addition of a base such as lithium / sodium hydroxide in aqueous solution between 15-40 ° C. (process 7).

The hydroxy-DHA esters are useful intermediates for introducing other functional groups at the α-position according to the invention. The hydroxyl group can be activated by conversion to a halide or tosylate before reacting with other nucleophilic materials such as ammonia, amine thiols and the like. The Mitsunobu reaction is also useful for converting hydroxyl groups to other functional groups (Mitsunobu, O, Synthesis, 1981, 1).

The compounds of formula (I) as defined above can be synthesized by combining the processes already mentioned. The present invention includes the aforementioned process.

The present invention provides a process for preparing a pharmaceutical composition comprising mixing a compound of formula (I), a pharmaceutically acceptable salt, solvate, complex or prodrug thereof with a pharmaceutically acceptable adjuvant, diluent or carrier. .

Pure compounds of the enantiomers can be prepared by decomposing racemic compounds of the formula (I) already mentioned. Decomposition of the compound of formula (I) is a well known decomposition method, such as, for example, reacting a compound of formula (I) with a purely adjuvant to make a diastereomeric mixture which can be separated by chromatography. It can be performed as. Two enantiomers of the compound of formula (I) can thus be generated from diastereomers by general methods such as hydrolysis.

As mentioned earlier in the α-position of DHA, stoichiometric chiral auxiliaries can be used for the asymmetric introduction effect of substituents. The use of chiral oxazolidin-2-one has proven methodologically of particular efficacy. Enolic acid derivatives of chiral N-acyloxazolidine can lower various electrophiles in a highly stereoregulated manner (Ager, Prakash, Schaad 1996).

The invention can be better understood through the following examples, which do not limit the scope of the invention. In the examples below, the structural formulas were verified by mass spectrometry (MS). It is also evident that fatty acid derivatives can be prepared from low or medium concentrations of DHA (ie about 40-60 W% DHA).

Synthesis protocol

α -methylDHA Preparation of EE ( PRB- I)

Butyl lithium (228 ml, 0.37 mol, 1.6 M in nucleic acid) was 0 ° C., N ₂ Under the atmosphere was added dropwise to diisopropylamine stirred solution (59.5 ml, 0.42 mol) in dry THF (800 ml). The result was stirred at 0 ° C. for 30 minutes, cooled to −78 ° C. before being slowly added to DHA EE (100 g, 0.28 mol) in dry THF (500 ml) for 2 hours and further stirred for 30 minutes. The dark green solution was stirred at −78 ° C. for 30 minutes before MeI (28 ml, 0.45 mol) was added. The solution was added to an aqueous solution (1.51) after reaching up to -20 ° C for 1.5 hours and extracted with heptane (2 x 800 ml). The combined organic phases were washed with 1M HCl (1 L), dried (Na ₂ SO ₄ ), filtered, and evaporated under reduced pressure. The result was eluted with heptane / EtOAc (99: 1) dry flash chromatography on silica gel to give 50 g (48%) of a compound as a pale yellow oil;

¹ H-NMR (200 MHz, CDCl ₃ ) δ1.02 (t, J7.5 Hz, 3H), 1.20 (d, J6.8 Hz, 3H), 1.29 (t, J 7.1 Hz, 3H), 2.0-2.6 (m , 5H), 2.8-3.0 (m, 10H), 4.17 (t, J7.1 Hz, 2H), 5.3-5.5 (m, 12H);

MS (electrospray); 393 [M + Na].

α- ethyl DHA Preparation of EE ( PRB- 2)

Butyl lithium (440 ml, 0.67 mol, 1.6 M in nucleic acid) is 0 ° C. N ₂ To the mixture was added dropwise to diisopropylamine stirred solution (111 ml, 0.78 mol) in dry THF (750 ml). The result was stirred at −78 ° C. for 45 minutes before being slowly added to DHA EE (200 g, 0.56 mol) in dry THF (1.6 l). Ester was added over 4 hours. The dark green solution was stirred at −78 ° C. for 30 minutes before adding EtI (65 ml, 0.81 mol). The solution reached -40 ° C before further addition of EtI (5 ml, 0.06 mol), and -15 ° C (-78 ° C over 3 hours) before the mixture was added to the aqueous solution and extracted with nucleic acid (2x). It was raised to). The combined organic phases were washed with 1M HCl, dried (Na ₂ SO ₄ ), filtered, and evaporated under reduced pressure. The result was eluted by flash chromatography on silica gel with heptane / EtOAc (from 99: 1 to 50: 1) to give 42.2 g (20%) of a yellow oil;

¹ H-NMR (200 MHz; CDCl ₃ ) δ 0.8-1.0 (m, 6H), 1.2-1.4 (m, 4H), 1.5-1.7 (m, 2H), 2.12 (m, 2H), 2.3-2.5 ( m, 2H), 2.8-3.0 (m, 10H), 4.18 (t, J7.1 Hz, 2H), 5.3-5.6 (m, 12H);

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

α- Ethoxy - DHA Ethyl ester  Produce( PRB -3)

Solution of α-hydroxyDHA ethyl ester (PRB-12) in 5 mL of THF (372 mg, 1.00 mmol) was added to a suspension of 60% NaH (84.1 mg, 2.1 mmol) in 5 mL of THF under -78 ° C N ₂ atmosphere. Dropwise, the mixture was stirred at −78 ° C. for 20 minutes, after which time ethyl iodide (0.24 mL, 3.01 mmol) was added slowly. The reaction compound was gradually warmed to room temperature overnight. 15 mL of saturated aqueous NH ₄ Cl was added, and the mixture was extracted with 25 mL × 2 of diethyl ether, and the organic phase was washed with 25 mL brine, dried (Na ₂ SO ₄ ), filtered, evaporated under reduced pressure, and then silica gel Flash chromatography on eluted with heptane / EtOAc (95: 5) to give 68 mg (17%) of a yellow liquid product.

¹ H NMR (200 MHz, CDCl ₃ ) δ0.94 (t, J = 7.5 Hz, 3H), 1.16-1.29 (m, 6H), 2.05 (quint, J = 7.2 Hz, 2H), 2.50 (m, 2H ), 2.76-2.84 (m, 10H), 3.33-3.48 (m, IH), 3.53- 3.71 (m, IH), 3.83 (dd, J = 6.8 Hz, J = 6.2 Hz, IH), 4.18 (q, J = I. \ Hz, 2H), 5.31-5.45 (m, 12H)

¹³ C NMR (50 MHz, CDCl ₃ ) δ14.2, 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 signals), 128.2 (2 signals) , 128.5, 130.7, 132.0, 172.5 (3 signals hidden)

MS (electrospray); 423 [M + Na] ⁺

α- Floro - DHA EE Manufacturing of PRB -4)

LDA (2.1 ml, 4.2 mol, 2 M in THF / heptane / ethylbenzene solvent) in dry THF (10 ml) under N ₂ atmosphere at −78 ° C., DHA EE (1 g, 2.8 in dry THF (30 ml)) mmol) dropwise for 15 minutes. NFSi (1.06 g, 3.4 mmol) was then added. The solution was warmed to room temperature and then stirred for 70 hours. The mixture was added to water and extracted with nucleic acid (2x). The mixed organic phase was washed with 1M HCl, washed with water, dried (Na ₂ SO ₄ ), filtered, and evaporated under reduced pressure; MS (electrospray); 397 [M + Na].

α-α-dimethyl DHA EE Manufacturing of PRB -5)

Butyl lithium (100 ml, 0.17 mol, 1.6 M in nucleic acid) was 0 ° C., N ₂ To the mixture was added dropwise to diisopropylamine stirred solution (28 ml, 0.20 mol) in dry THF (100 ml). The resulting solution was stirred at 0 ° C. for 30 minutes, cooled to −78 ° C. and DHA EE (50 g, 0.14 mol) in dry THF was added slowly. Before adding MeI (17 ml, 0.28 mol), the resulting dark green solution was stirred for 30 min at -78 ° C. The solution was raised to −10 ° C., then added to water and extracted with nucleic acid (2 ×). The mixed organic phase was washed with 1M HCl, dried (Na ₂ SO ₄ ), filtered and evaporated under reduced pressure.

The procedure was repeated except that α-methyl DHA EE was used instead of DHA EE. The material eluted dry flash chromatography on silica gel with heptane / EtO Ac (until 99: 1 became 98: 2) to give 31.6 g (59%) of a pale yellow oil.

¹ H-NMR (200 MHz; CDCl ₃ ) δ1.01 (t, J7.5 Hz, 3H), 1.21 (s, 6H), 1.28 (t, J7.1 Hz, 3H), 2.08 (m, 2H) 2.34 (d, J 6.8 Hz, 2H), 2.8-3.0 (m, 10H), 4.15 (q, J7.5 Hz ₅ 2H), 5.3-5.6 (m, 12H);

¹³ C-NMR (50 MHz; CDCl ₃ ) δ14.7, 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); 385 [M + H].

α- Thiomethyl Of DHA  Produce( PRB -6)

MeSNa (80 mg, 1.14 mmol) was dissolved in 20 mL THF in α-Iodo DHA EE (0.5 g, 1.04 mmol) at 0 ° C. N ₂ Added under atmosphere, the mixture was stirred for several minutes before diluting in heptane. The organic phase was washed with water (2 ×), dried (Na ₂ SO ₄ ), and evaporated under reduced pressure. Flash chromatography gave a light yellow oil of α-thiomethyl DHA EE. The α-thiomethyl DHA EE was dissolved in 10 mL EtOH and 10 mL THF. The solution was added to LiOH (0.39 g, 9.2 mmol) dissolved in 5 mL water. The reaction mixture was stirred overnight at room temperature before dilution with water and heptane. The organics were extracted with IM LiOH (2 ×) and the mixed aqueous phase was acidified with 5M HCl and extracted with diethyl ether (2 ×). The mixed organic phase is washed with brine and water, dried (Na ₂ SO ₄ ), and evaporated under reduced pressure. 183 mg (47%) of a compound as a pale yellow oil were obtained;

¹ H-NMR (200 MHz, CDCl ₃ ) δ 0.98 (t, J6.6 Hz, 3H), 1.95-2.65 (m, 7H), 2.72-3.55 (m, 10H), 3.12-3.43 (m, IH), 5.20-5.70 (m, 12 H), 10.65 (br s, IH);

¹³ H-NMR (50 MHz, CDCl ₃ ) δ14.7, 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.

α- Thioethyl DHA EE Manufacturing of PRB -7)

EtSNa (2.1 g, 25 mmol) was added to α-iodo DHA EE (11 g, 23 mmol) dissolved in 100 mL of THF under O ° C. N ₂ atmosphere and the reaction was stirred at 0 ° C. for one hour. The reaction was terminated by addition with IM HCl and diluted with heptane. The organic phase was washed with water (2 ×), dried (Na ₂ SO ₄ ) and evaporated under reduced pressure. Flash chromatography was eluted with heptane / EtOAc (30: 1) to give 7.3 g (76%) of a compound as a pale yellow oil;

¹ H-NMR (200 MHz, CDCl ₃ ) δ1.1-1.3 (m, 9H), 2.05 (m, 2H), 2.3-2.7 (m, 4H), 2.7-2.9 (m, 10H), 3.25 (m, IH), 4.17 (q, J7.1 Hz, 2H), 5.3-5.5 (m, 12H);

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

α-α- Diethyl DHA EE Manufacturing of PRB -8)

Butyl lithium (38.6 ml, 0.62 mol, 1.6 M in nucleic acid) was 0 ° C., N ₂ To the mixture was added dropwise to diisopropylamine stirred solution (9.1 ml, 0.65 mol) in dry THF (100 ml). The resulting solution was stirred at 0 ° C. for 30 minutes, cooled to −78 ° C. and then slowly added to DHA EE (20.0 g, 0.56 mol) in dry THF (100 ml). Before adding EtI (6.8 ml, 0.84 mol), the resulting dark green solution was stirred for 30 min at -78 ° C. The solution was raised to −10 ° C. and then added to water and extracted with nucleic acid (2 ×). The mixed organic phases were washed with 1 M HCl, dried (Na ₂ SO ₄ ), purified and evaporated under reduced pressure.

The procedure was repeated except that α-ethyl DHA EE was used instead of DHA EE. After addition of EtI, the reaction mixture was warmed to room temperature and stirred overnight. The product was eluted with dry flash chromatography on silica gel with heptane / EtO Ac (until 99: 1 became 98: 2) to give 10.0 g (43%) of a pale yellow oil.

¹ H-NMR (200 MHz; CDCl ₃ ) δ0.83 (t, J 7.4 Hz, 6H), 0.94 (t, J 5.8 Hz, 3H), 1.28 (t, J 7.1 Hz, 3H), 1.63 (q, J 7.4 Hz ₅ 4H), 2.10 (m, 2H), 2.34 (d, J 6.9 Hz, 2H), 2.8-3.0 (m, 10H), 4.15 (q, J7.5 Hz, 2H), 5.3-5.6 ( m, 12H);

¹³ C-NMR (50 MHz; CDCl ₃ ) δ8.9, 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); 413.3 [M + H], 435.3 [M + Na].

α-benzyl DHA EE Manufacturing of PRB -9)

n-BuLi (1.6 M in nucleic acid, 3.86 mL, 6.18 mmol) was added dropwise to a stirred solution of diisopropyl amine (0.91 mL, 6.46 mmol) in dry THF (20 mL) under O 0 C inert atmosphere. The mixture was stirred at 0 ° C. for 30 minutes and at 78 ° C. for 5 minutes. DHA EE (2.0 g, 5.62 mmol) in dry THF (10 mL) was added slowly. The mixture was stirred at −78 ° C. for 20 minutes before benzyl bromide (0.80 mL, 6.74 mmol) was added. The resulting solution was warmed to O ° C. over 3 hours and separated into water (100 mL) and heptane (100 mL). The aqueous layer was extracted with heptane (50 mL), and the mixed organic layer was washed with IM HCl and dried (Na ₂ SO ₄ ). Concentrated under reduced pressure, then purified by flash chromatography (heptane: EtOAc 99: 1) to 1.05 g (42%) to give a compound of a colorless oil;

¹ H-NMR (200 MHz, CDCl ₃ ): δ 0.99 (t, 3H), 1.18 (t, 3H), 2.08-2.16 (m, 2H), 2.35-2.42 (m, 2H), 2.74-2.98 (m , 13H), 4.09 (q, 4H), 5.38-5.50 (m, 10H), 7.19-7.36 (m, 5H);

¹³ C-NMR (50 MHz, CDCl ₃ ): 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): 447.3 [M + H], 469.3 [M + Na].

α- EthanesulfinylDHA EE Manufacturing of PRB -10)

-20 ℃ under an inert atmosphere in 15 mL CHCl ₃ α- thioethyl EE DHA (0.5 g, 1.3 mmol) in 10 mL CHCl ₃ was added MCPBA (0.22 g, 1.3 mmol) the solution became more. The reaction mixture was stirred at this temperature for 2 hours, filtered and washed with saturated aqueous NaHCO ₃ solution. The aqueous phase was extracted twice with CHCl ₃ , and the mixed organic phases were washed with water and brine, dried (Na ₂ SO ₄ ), filtered and concentrated. The material was subjected to flash chromatography using nucleic acid: EtOAc (8: 2) to obtain 0.35 g (70%) of the compound.

¹ H NMR (200 MHz, CDCl ₃ ): δ 0.99 (t, 3H), 1.27-1.45 (m, 6H), 2.09 (m, 2H), 2.79-2.94 (m, 14H), 3.55 (m, IH) , 4.25 (q, 2H), 5.37-5.59 (m, 12H).

¹³ C NMR (50 MHz ₅ CDCl ₃ ): 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): 455.3 [M + Na].

α- Thio phenyl - DHA Ethyl ester  Produce( PRB -11)

To a solution of α-iodo-DHA ethyl ester (PRB-15) (3.40 g, 7.05 mmol) in 20 mL of acetone was added dropwise with a solution of 1.039 g, 7.86.mmol of sodium phenyl sulfide in 110 mL acetone. It became. The resultant was stirred at room temperature for 1 hour 30 minutes, and evaporated under reduced pressure, followed by flash chromatography on silica gel using heptane / EtOAc 200: 1-95: 5 to give 2.35 g (72) of a yellow liquid compound. %) Was obtained.

¹ H NMR (200 MHz, CDCl ₃ ) δ 0.97 (t, J = 7.5 Hz, 3H), 1.18 (t, J = 7.1 Hz, 3H) ₅ 2.09 (quint, J = 7.1 Hz, 2H), 2.54- 2.66 (m, 2H), 2.83-2.86 (m, 10 H), 3.67 (dd, J = 6.8 Hz, J = 8.3 Hz, 1 H), 4.12 (q, J = 7.1 Hz, 2H), 5.24-5.49 (m, 12H), 7.28-7.33 (m, 3H), 7.46-7.50 (m, 2H)

¹³ C NMR (50 MHz, CDCl ₃ ) δ14.0, 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 signals hidden)

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

HRMS (EI) calculated for C ₃₀ H ₄₀ O ₂ S: 464.2749, found: 464.2741

α-hydroxy- DHA Ethyl ester  Produce( PRB -12)

1.6 M BuLi in nucleic acid (87.5 mL, 140 mmol) was added dropwise to a solution of diisopropyl amine (19.76 mL, 140 mmol) in 40 mL of dry THF at −78 ° C., N ₂ atmosphere. The resulting mixture was stirred at −78 ° C. for 15 minutes before a solution of DHA ethyl ester (24.99 g, 70.1 mmol) in 80 mL of THF was added slowly. The dark green resultant reaction mixture was stirred at −78 ° C. for 1 hour before triethylphosphate (12.2 mL, 70.1 mmol) was added slowly, after which the reaction mixture was kept at −78 ° C. for 5 hours and then slowly returned to room temperature. O ₂ was passed through the reaction mixture overnight while raising the temperature.

100 mL of saturated aqueous NaHCO ₃ was added and the mixture was extracted with 200 mL × 2 of diethyl ether. The organic phase was dried over Na ₂ SO ₄ , filtered, and evaporated under reduced pressure, followed by flash chromatography on silica gel using heptane / EtOAc 99: 1 to 95: 5 to give 4.52 g of a yellow liquid compound ( 17%).

¹ HNMR (200 MHz, CDCl ₃ ) δ 0.92 (t, J = 7.5 Hz, 3H), 1.24 (t, J = 7.1 Hz, 3H), 5 2.02 (quint, J = 7.1 Hz, 2H), 2.44-2.54 (m, 2H), 2.74-2.87 (m, 10H), 4.13-4.24 (m, 3H), 5.25-5.94 (m, 12H)

¹³ CNMR (50 MHz, CDCl ₃ ) δ14.0, 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 signals hidden)

MS (electrospray); 395 [MH-Na] ⁺

HRMS (ES) calculated for C ₂₄ H ₃₆ O ₃ Na: 395.2556, found: 395.2543

α- methyl - DHA  Preparation of amides PRB -13)

0.1 mL of DMF is added to a solution of α-methyl-DHA (PRB-I FA) (3.13 g, 9.1 mmol) and oxaryl chloride (8.0 mL, 94.5 mmol) in 90 mL of toluene, and the resulting mixture is N at room temperature. _It stirred for 15 hours and 30 minutes in ₂ atmosphere. The mixture was evaporated under reduced pressure and the residue was dissolved in 100 mL of THF, cooled to 0 ° C., and aqueous solution NH ₃ (20 mL) was added slowly. The ice bath was removed and the mixture was stirred at room temperature for 4 hours, 50 mL more water was added and the aqueous phase was extracted with 2 × 100 mL diethyl ether. The organic phase was washed with 50 mL of saturated aqueous NH ₄ Cl, dried over Na ₂ SO ₄ , filtered, and evaporated under reduced pressure followed by CH ₂ C1 ₂ / 2M NH _{3 in} MeOH. Purification by flash chromatography on silica gel using 97.5: 2.5 gave 2.51 g (80%) of a yellow liquid compound.

¹ H NMR (200 MHz, CDCl ₃ ) δ0.91 (t, J = 7.5 Hz, 3H), 1.10 (d, J = 9.8 Hz, 3H), 1.94-2.11 (m, 3H), 2.19-2.35 (m, 2H), 2.76-2.77 (m, 10 H), 5.18-5.45 (m, 12 H), 6.03 (s, IH), 6.72 (s, IH)

¹³ C NMR (50 MHz, CDCl ₃ ) δ14.6, 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 signals hidden)

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

HRMS (EI) calculated for C ₂₃ H ₃₅ NO: 341.2719, found: 341.2707

α- Methoxy - DHA Ethyl ester  Produce( PRB -14)

A solution of α-hydroxy-DHA ethyl ester (PRB-12) (373 mg, 1.00 mmol) in 5 mL of THF was -78 ° C., N ₂ Under atmosphere was added to a suspension of 60% NaH (61.1 mg, 1.53 nimol) in 5 mL of dry THF, and stirred at −78 ° C. for 20 minutes before slowly adding methyl iodide (0.13 mL, 2.09 mmol) to the resulting mixture. . The reaction mixture was gradually warmed up to room temperature for 5 hours. 15 mL of saturated aqueous NH ₄ Cl was added and the mixture was extracted with 25 mL × 2 of diethyl ether and the organic phase was washed with 25 mL of brine, dried over Na ₂ SO ₄ , filtered, and evaporated under reduced pressure and then heptane / EtOAc Purification by flash chromatography on silica gel using 99: 1-4: 1 gave 136 mg (35%) of a yellow liquid compound.

¹ H NMR (200 MHz, CDCl ₃ ) δ0.92 (t, J = 7.5 Hz, 3H), 1.24 (t, J = 7.1 Hz, 3H), 2.03 (quint, J = 7.3 Hz, 2H), 2.48 (t, J = 5.7 Hz, 2H), 2.73-2.82 (m, 10H), 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)

¹³ C NMR (50 MHz, CDC ₁₃ ) δ14.1, 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 signals hidden)

MS (electrospray); 409 [M + Na] ⁺

HRMS (ES) calculated for C ₂₅ H ₃₈ O ₃ Na: 409.2713, found: 409.2711

α- IodoDHA EE Manufacturing of PRB -15)

Diisopropylamine (20 mL, 140 mmol) was dissolved in 150 mL THF. n-BuLi (88 mL, 140 mmol, 1.6 M) was added slowly under -20 ° C N ₂ atmosphere and the solution was cooled to -78 ° C. DHA EE (50 g, 140 mmol) in 250 mL THF was added slowly to the solution and the reaction mixture was stirred at room temperature for 30 minutes. The resulting mixture was slowly added to a solution of I ₂ (42.8 g, 169 mmol) in 400 mL THF under −78 ° C. N ₂ atmosphere. The reaction was terminated by IM HCl addition and diluted with heptane. The organic phase was washed with 10% Na ₂ S ₂ O ₃ (2 ×), dried over Na ₂ SO ₄ , filtered, and evaporated under reduced pressure. Flash chromatography on silica gel using heptane / EtOAc (100: 1) gave 11.0 g (16%) of a pale yellow oil;

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

α- Iodo - DHA Ethyl ester  Produce( PRB -15)

1.6 M BuLi in nucleic acid (158 mL, 253 mmol) was -78 ° C N ₂ Under atmosphere was added dropwise to a solution of diisopropyl amine (42 mL, 298 mmol) in 150 mL of dry THF. The result was stirred for 35 minutes at −78 ° C. before a solution of DHA ethyl ester (75.05 g, 210 mmol) in 300 mL of THF was added slowly. The dark green reaction mixture was stirred at −78 ° C. for 30 minutes before the addition of a solution of I ₂ (91.06 g, 359 mmol) in 200 mL of THF. The reaction product was stirred at −78 ° C. for 20 minutes, then the reaction was terminated by adding 200 mL of water and extracted with 300 mL of heptane. The organic phase was washed with 1 M HCl, 150 mL and 200 mL of water, dried over Na ₂ SO ₄ , filtered, and evaporated under reduced pressure. The resulting material was subjected to flash chromatography on silica gel using heptane / EtOAc (100: 1) to give 26.14 g (26%) of a yellow solution;

¹ H NMR (200 MHz, CDCl ₃ ) δ 0.94 (t, J = 7.5 Hz, 3H), 1.24 (t, J = 7.1 Hz, 3H). 2.04 (quint, J = 7 Hz ₃ 2H), 2.69-2.84 (m, 12H), 4.17 (q, J = 7.1 Hz, 2H), 4.22 (t, j = 7.9 Hz, IH), 5.24-5.49 ( m, 12 H)

¹³ C NMR (50 MHz, CDCl ₃ ) δ13.7, 14.2, 25.5, 26.0 (2 signals), 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 signals hidden)

MS (electrospray); 505 [M + Na] ⁺

α-amino- DHA Ethyl ester  Produce( PRB -17)

Hydrazine hydrate (46 μl, 0.95 mmol) is added to a solution of α-phthalimide-DHA ethyl ester (313.5 mg, 0.62 mmol) in 5 mL of EtOH and the mixture is refluxed under N ₂ atmosphere for 15 h 30 min. 149 mg (64%) of a yellow liquid compound were evaporated under reduced pressure and subjected to silica gel flash chromatography using CH ₂ C1 ₂ : 7M NH ₃ in MeOH (99: 1-95: 1). Got it.

¹ H NMR (200 MHz, CDCl ₃ ) δ 0.91 (t, J = 7.5 Hz, 3H), 1.22 (t, J = 7.1 Hz, 3H), 1.72 (bs, 2H), 2.02 (quint., J = 7.2 Hz, 2H), 2.39-2.46 (m, 2H), 2.73-2.82 (m, 10 H), 3.47 (bs, IH), 4.13 (q, 2H) ₅ 5.23-5.56 (m, 12 H)

¹³ C NMR (50 MH _Z5 CDCl ₃ ) δ14.1, 20.4, 25.4, 25.5, 25.6, 54.1, 60.8, 124.4, 126.9, 127.7 (2 signals), 127.9, 128.2, 128.3, 128.4, 131.4, 131.9, 189.3 (6 signals hidden)

MS (electrospray); 372 [M + H] ⁺

(S)-(+)-α-ethyl DHA EE Manufacturing of PRB -20)

Synthesis of Intermediates PRB-18:

DHA (3.00 g, 18.3 mmol) was dissolved in dry CH ₂ Cl ₂ (120 mL) under an O ° C. inert atmosphere and DMAP (2.45 g, 20.1 mmol) and DCC (3.96 g, 19.2 mmol) were added. The mixture was stirred at O &lt; 0 &gt; C for 20 minutes, and (4R, 5S)-(+)-4-methyl-5-phenyl-2-oxazolidinone (3.24 g, 18.3 mmol) was added and then 20 hours at room temperature. Was stirred. Purification by filtration and flash chromatography (heptane: EtOAc 6: 1) gave 3.00 g (34%) of a PRB-18 intermediate of a colorless oil.

¹ H-NMR (200 MHz ₅ CDCl ₃ ): δ 0.93-1.05 (t + d, 6H), 2.11 (m, 2H), 2.51 (m, 2H), 2.80-3.00 (m, 10H), 3.05 ( m, 2H), 4.77 (m, IH), 5.34-5.68 (m, 12H), 5.70 (d, IH), 7.28.7.32 (m, 2H), 131-1Al (m, 3H).

Synthesis of Intermediates PRB-19:

PRB-18 (1.80 g, 3.70 mmol) in dry THF (10 mL) was dropped in a solution of LiHMDS (IM in THF, 4.00 mL, 4,00 mmol) in dry THF (15 mL) at -78 ° C. under inert atmosphere. Added freshly. The mixture was stirred at −78 ° C. for 30 minutes, EtI (0.89 mL, 11.1 mmol) was added and the temperature was slowly raised to O ° C. over 1 hour. The mixture was stirred for 18 h at 0 ° C. and separated into saturated NH ₄ Cl (50 mL) and diethyl ether (50 mL) layers. The aqueous layer was extracted with diethyl ether (50 mL) and the combined organic layers were washed with 0.1 M HCl (50 mL) and brine (50 mL). Purification using dry (Na ₂ SO ₄ ) and flash chromatography (heptane: EtOAc 95: 5) gave 0.52 g (27%) of the PRB-19 intermediate as a colorless oil.

¹ H NMR (200 MHz, CDCl ₃ ): δ0.88-1.01 (m, 9H), 1.64-1.78 (m, 2H), 2.08 (m, 2H), 2.31 (m, IH), 2.48 (m, IH ), 2.87 (m, 10H), 3.87 (m, IH), 4.75 (m, IH), 5.32 (m, 12H), 5.63 (d, J7.1 Hz, IH), 7.32 (m, 2H), 7.42 (m, 3 H).

¹³ C NMR (50 MHz, CDCl ₃ ): 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): 538.2 [M + Na].

PRB-19 (0.25 g, 0.485 mmol) was dissolved in absolute EtOH (5 mL) at 0 ° C. under inert atmosphere. NaOEt (IM in EtOH, 0.54 mL, 0.54 mmol) was added, stirred at O ° C. for 30 min, and divided into water and heptane layers. The aqueous layer was extracted with heptane and the combined organic layers were washed with 0.1M HCl and dried. Purification by flash chromatography yielded 0.025 g (13%) of the compound PRB-20 as a colorless oil.

¹ H NMR (200 MHz; CDCl ₃ ) δ 0.8-1.0 (m, 6H), 1.2-1.4 (m, 4H), 1.5-1.7 (m. 2H), 2.12 (m, 2H), 2.3-2.5 ( m, 2H), 2.8-3.0 (m, 10H), 4.18 (t, 2H), 5.3-5.6 (m, 12H).

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

[α] _D + 1.7 ° (c = 1.5, ethanol).

(R)-(-)-αethyl DHA EE Manufacturing of PRB -23)

Synthesis of Intermediates PRB-21:

DHA (1.0Og, 3.05 mmol) was dissolved in dry CH ₂ Cl ₂ (20 mL) under O inert atmosphere, and DMAP (0.41 g, 3.35 mmol) and DCC (0.66 g, 3.20 mmol) were added. The mixture is stirred at O &lt; 0 &gt; C for 20 minutes, (4S, 5R)-(-)-4-methyl-5-phenyl-2-oxazolidinone (0.54 g, 3.05 mmol) is added and at room temperature for 20 hours Stirred. Purification using filtration and flash chromatography (heptane: EtOAc 6: 1) gave 1.08 g (73%) of the PRB-21 intermediate as a colorless oil.

¹ H-NMR (200 MHz, CDCl ₃ ): δ 0.93-1.05 (t + d, 6H), 2.11 (m, 2H), 2.51 (m, 2H), 2.80-3.00 (m, 10H), 3.05 ( m, 2H), 4.77 (m, IH), 5.34-5.68 (m, 12H), 5.70 (d, IH), 7.28.7.32 (m, 2H), 131-1 Al (m, 3H).

Synthesis of Intermediates PRB-22:

PRB-21 (3.25 g, 6.67 mmol) in dry THF (15 mL) was dropped in LiHMDS (IM in THF, 7.34 mL, 7,34 mmol) solution in dry THF (35 mL) under an inert atmosphere at -78 ° C. Added. The mixture was stirred at −78 ° C. for 30 minutes, EtI (1.6 mL, 20.0 mmol) was added and the temperature was slowly raised to O ° C. over 1 hour. The compound was then stirred for 18 h at 0 ° C. and partitioned into NH ₄ Cl (50 mL) and diethyl ether (50 mL) layers. The aqueous layer was extracted with diethyl ether (50 mL), and the combined organic layers were washed with 0.1M HCl (50 mL) and brine (50 mL). Purification using dry (Na ₂ SO ₄ ) and flash chromatography (heptane: EtOAc 95: 5) gave 1.50 g (44%) of the intermediate PRB- 22 as a colorless oil.

¹ H NMR (200 MHz, CDCl ₃ ): δ0.88-1.01 (m, 9H), 1.64-1.78 (m, 2H), 2.08 (m, 2H), 2.31 (m, IH), 2.48 (m, IH ), 2.87 (m, 10H), 3.87 (m, IH), 4.75 (m, IH), 5.32 (m, 12H), 5.63 (d, J7.1 Hz, IH), 7.32 (m, 2H), 7.42 (m, 3 H).

¹³ C NMR (50 MHz, CDCl ₃ ): 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): 538.2 [M + Na].

PRB-22 (0.25 g, 0.485 mmol) was dissolved in absolute EtOH (5 mL) at 0 ° C. inert atmosphere. NaOEt (IM in EtOH, 0.54 mL, 0.54 mmol) was added and the mixture was stirred at 0 ° C. for 30 min and divided into water and heptane layers. The aqueous layer was extracted with heptane and the combined organic layers were washed with 0.1M HCl (50 mL) and dried. Purification using flash chromatography (heptane: EtOAc 95: 5) gave 0.025 g (13%) of the compound PRB-23 as a colorless oil.

¹ H-NMR (200 MH _Z ; CDCl ₃ ) δ0.8-1.0 (m, 6H), 1.2-1.4 (m, 4H), 1.5-1.7 (m, 2H), 2.12 (m, 2H), 2.3- 2.5 (m, 2H), 2.8-3.0 (m, 10H), 4.18 (t, 2H) ₃ 5.3-5.6 (m, 12H);

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

[α] _D -1.3 ⁰ (c = 1.00, ethanol).

α- Phthalimide - DHA Ethyl ester  Produce( PRB -24)

A mixture of α-hydroxy-DHA ethyl ester (PRB-12) (373.5 mg, 1.00 mmol), phthalimide (178 mg, 1.21 mmol) and triphenyl phosphine (313.9 mg, 1.20 mmol) in 10 mL THF was N After cooling to 0 ° C. under ₂ atmospheres, diisopropylazodicarboxylate (0.24 mL, 1.24 mmol) was added dropwise. The cooling bath was removed and the reaction mixture was stirred at room temperature for 18 hours, then evaporated under reduced pressure and subjected to silica gel flash chromatography using heptane / EtOAc (99: 1-95: 1) to give a yellow solution of the compound 323 mg (64%) was obtained.

¹ H NMR (200 MHz, CDCl ₃ ) δ 0.95 (t, J = 7.5 Hz, 3H), 1.22 (t, J = 7.1 Hz, 3H), 2.05 (m, 2H), 2.72-2.84 (m, 11H ), 3.02-3.22 (1H), 4.20 (q, J = 7.1 Hz, 2H), 4.87 (dd, J = I l Hz, J = 4.9 Hz ₅ IH), 5.17-5.40 (m, 12H), 7.68- 7.75 (m, 2H), 7.79-7.85 (m, 2H)

¹³ C NMR (50 MHz ₅ CDCl ₃ ) δ14.0, 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 signals hidden)

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

α- Ethylamino - DHA Ethyl ester ( PRB -25) and α- Diethylamino - DHA Ethyl ester  Preparation of (PRB-26)

To a mixture of α-amino-DHA ethyl ester (PRB-17) (746.5 mg, 2.01 mmol), LiOH-H ₂ O (171.6 mg, 4.09 mmol) and 4 μg (599 mg) of molecular sieve in 4 mL of DMF, ethylbromide ( 3.0 ml, 40.2 mmol) was added and the resulting mixture was stirred at room temperature for 71 hours. The mixture was diluted with 100 mL of diethyl ether and filtered. The organic phase was washed with 20 mL of 1M NaOH _{5 and} 20 mL of brine, dried (Na ₂ SO ₄ ), and evaporated under reduced pressure, then heptane: EtOAc (95: 5) -CH ₂ C1 ₂ : MeOH (99: 1): Silica gel flash chromatography was performed using 2M NH ₃ to obtain 458 mg (53%) of PRB-26 in the yellow solution and 152 mg (19%) of PRB-25 in the yellow solution.

PRB-26:

¹ H NMR (200 MHz, CDCl ₃ ) δ 0.99 (t, J = 7.5 Hz, 3H), 1.03 (t, 3H), 1.24 (t, J = 7.1 Hz, 6H), 2.05 (quint, J = 7.1 Hz, 2H), 2.52 (m, 4H), 2.76-2.85 (m, 12H), 3.35 (t, IH), 4.13 (q, J = 7.1 Hz, 2H), 5.28-5.44 (m, 12H )

¹³ C NMR (75 MHz, CDCl ₃ ) δ14.1, 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 signals hidden)

Example

2 shows an overview of the models and methods used to explain the effects of metabolic syndrome and type 2 diabetes in the present invention. Five blocks of experiments have been carried out to explain the effects of reducing DHA derivatives on insulin resistance and / or alleviating metabolic syndrome. The invention is not limited to the examples shown.

Example  1. In liver cells, intracellular free fatty acids ( Non-esterification  Analysis of fatty acids) 2  Block 1)

background

In the first block of the experiment (see Figure 2), liver tissues of animals administered PRB-1, 2, 5 and 7 were analyzed for free nonesterified fatty acids. The animals were supplemented in the fifth block of the experiment (pharmacodynamic effects of DHA derivatives in animal models of metabolic syndrome). The animals were given DHA (fat content of diet 15%) or DHA derivative (fat content of diet 1.5%) for 8 weeks and the stable levels of intracellular DHA and DHA derivatives were to be in steady state. Liver tissue was chosen because of the fact that the metabolic rate was very high in the liver.

Way

Liver samples were homogenized in cold PBS buffer and immediately extracted with chloroform: methanol (2: 1) containing 0.2 mM butylhydroxytoluene (BHT) using cis-10-heptadiseinoic acid as an internal standard. The organic phase was dried under nitrogen and dissolved again in acetonitrile containing 0.1% acetic acid and 10 μM BHT for RP-HPLC MS / MS analysis. Total protein content was measured using Bio-Rad method after homogenization.

An Agilent 1100 system was used to separate DHA and its PRB derivatives in reverse phase (Supelco Ascentis C _1S column, 25 cm × 4.6 mm, id 5 μM) within 22 minutes. The fluidized bed was acetonitrile-H ₂ O (87 + 13, v / v) of the same degree of change containing 0.1% acetic acid. The column oven temperature was set at 35 ° C. The column elution was identified and quantified by cathodic electrospray ionization using a multi-reaction monitoring mode with triple tandem quadrapole mass / mass (ABI Qtrap-4000). The parent-daughter ion pairs were 327.3 / 327.3 (DHA), 341.3 / 341.3 (PRB-I), 355.3 / 355.3 (PRB-2 and PRB-5), 387.3 /387.3 (PRB-7), 267.2 under unit analysis, respectively. /267.2 (LS. FA 17: 1). The residence times of the unit vowels were all 100 msec except for FA 17: 1 being 200 msec. Accurate demonstration of the isomeric PRB compound was made by the combination of retention time and characteristic mass / charge ratio. Second-order regression standard curves were used for quantification after internal standard calibration.

result

Different concentrations of DHA-derivatives and concentrations of DHA are expressed in μg per g of total amount of hepatocyte protein. 3 shows the concentration of different PRBs from animals receiving PRB-I, 2, 5 and 7 at a concentration of 1.5% of the total fat content in a high fat diet.

The intracellular concentration of the highest PRBs was PRB-2. In addition, PRB-5 showed a high concentration in cells but not as much as PRB-2. This discovery was unexpected.

Figure 4 shows the intracellular concentrations of DHA in the liver tissues of animals receiving different PRBs. DHA was significantly higher in animals receiving PRB-7 compared to the other three DHA derivatives. Animals receiving PRB-2 showed low DHA concentrations. It seems that PRB-7 has reconverted to DHA within a certain range.

PRB-2 had the highest intracellular concentration. This means that PRB-2 can religate to nuclear receptors (a form that can be converted into a pharmacological effect on the handling of blood sugar and fat).

Example  2

Computer based affinity test (block 2 in FIG. 2)

background

Nuclear receptors have been arranged and the amino acid sequence for PPARs and other related receptors involved in the genetic regulation of glucose and fat is known. X-ray crystallography and NMR spectroscopy of PPAR receptors are possible and computer-based affinity testing of fatty acids that ligand to receptors can be used to measure binding kinetics. This binding geometry, called binding mode or binding pose, involves the positioning of ligands to receptors and the structural state of ligands and receptors. Effective ligand docking can thus be analyzed.

Ligand affinity for a receptor is defined by two different parameters: docking of the ligand at the binding site of the receptor and electrostatic bonding of the carboxy groups or side branches of the head of specific amino acids and fatty acids of the receptor. .

As already known, PPARα receptors are more irregular compared to PPARγ, which means that PPARα accepts more fatty acids as ligands when compared to PPARγ. However, metabolic syndrome or type 2 diabetics are mainly obese, overweight, and have pathological blood fats, so the high triglycerides and low HDL-chol activation of PPARα receptors are important. Ideal drugs for the treatment of metabolic syndrome or type 2 diabetes should act as ligands for both receptors and preferably have high affinity for PPARγ receptors.

Way

The ranking according to the binding affinity of the different DHA derivatives was calculated and given the lowest binding affinity (LBA) and average binding affinity (ABE). 15 DHA derivatives (PRB-I to PRB-15) were tested by the computerized docking method. Some derivatives such as PRB-I, PRB-2, PRB-7, PRB-9, PRB-IO, PRB-11, PRB-12, PRB-13, PRB-14, and PRB-15 are r and s enantiomers. Exists, both of which were tested. The r and s PPAR ligands rosiglitazone and pioglitazone were also tested for control. These compounds have been registered as drugs for the treatment of diabetes.

result

The results are shown in Table 1, where Table 1 is the lowest binding energy (LBE) of the unit confirmation, the average binding energy (ABE) of the immediately placed confirmation (ABE) and the ICM of the tested compound. The fraction of the immediately placed confirmation of the low energy fbound of ICM-saved 20 is shown as a parameter. Affinity to RXRα was tested in the same manner. The RXRα receptor interacts with PPAR receptors that form heterodimers by ligands of fatty acids.

5 shows binding affinity for PPARγ receptors and PPARγ receptors are primarily involved in the transcription of proteins that regulate blood glucose. Clearly, both the PRB-2 r and s stereoisomers had high affinity for the PPARγ receptor. PRB-5 was slightly lower while PRB-8 had the highest ABE. These findings were exceptional and could be understood as more effective transcription of PPARγ activated genes, each of which regulates blood glucose.

FIG. 6 shows binding affinity for PPARα receptors primarily involved in metabolism of fat, plasma lipids, adipose tissue biology and weight control. Some DHA derivatives showed high binding affinity and PRB-8 had the highest value. This was also exceptional.

7 shows nuclear receptor RXRα binding affinity. The physiological effects of binding to the RXRα receptor have not been firmly established. It is known that RXR binds to PPAR receptors, thereby forming heterodimers and consequently initiate transcription of defined genes.

[Table 1]

ND = Not docked, c = the double bonds in all-cis form, r = R enantioisomer, s = S enantioisomer, ROSI = Rosiglitazone, PIO = Pioglitazone

Some PRBs have high LBE and ABE values for PPARα and PPARγ receptors compared to the parent compound DHA, as well as high LBE and ABE for rosiglitazone and pioglitazone, r and s PPAR ligands. The value was shown. Several PRBs indicate a potential competition between known antidiabetic rosiglitazone and pioglitazone. Ethyl derivatives of the alpha position of the same fatty acids did not increase affinity for both r and s types. In particular, PPARγ receptors were added. As mentioned before, PPARα receptors are long continuous bonds of more irregular fatty acids.

In conclusion, many DHA derivatives tested demonstrated affinity for PPARα and PPARγ receptors with better binding affinity than rosiglitazone and pioglitazone.

Example  3

Affinity test in infected cells (block 3 of FIG. 2)

background

The release of luciferases correlates with the transcription of genes. Ligand binding to nuclear receptors such as PPARγ releases luciferase, including transcription of each gene. This technique provides a measure of the activity of the causal gene as well as the ligand affinity for the receptor.

Way

Transient infection of COS-I cells was carried out in 6-well plates as described in Graham and van der Eb (Graham). For the whole PPAR infection study, each well was given a 5 μg reporter structure, 2.5 μg pSV-β galactosidase, 0.4 μg pSG5-PPARγ2 as an internal control. The cells were harvested after 72 hours and luciferase activity was measured according to the protocol (Promega). Luciferase activity was normalized to β-galactosidase activation. Adipocytes were infected with D11 of differentiation using 16 μl of lipofectamine plus reagent, 4 μl of lipofectamine (Life Technologies Inc.), 0.2 μg of pSG5-PPARγ and 100 ng of pTK Renilla luciferase as a control of infection rate. Three hours after infection, cells were cultured in serum containing medium and incubated for 48 hours in the same medium containing the appropriate agent. Luciferase activity was determined by the method recommended by the manufacturer (Dual Luciferase assay, Promega). All infections were performed in sets of three.

Fatty acids (BRL or DHA) and PRB (mother liquor) were dissolved to a final concentration of 0.1 M in DMSO. The fat was then dissolved at 10 mM in DMSO and stored at −20 ° C. in a 1.5 m tube (Homoplimer, plastic tube) fed with argon. 10 μM of PRB or fatty acid and DMSO (control) were added to the medium 5 hours after infection. Infected cells were preserved for 24 hours before being lysed in the reporter lysis buffer. The binding of PRB or fatty acid to LBD of PPAR activates the binding of GAL4 to UAS and stimulates the tk promoter to express luciferase. Luciferase activity was measured using a luminometer (TD-20 / 20 luminometer; Turner Designs, Sunnycvale, Calif.) And normalized to protein content.

result

8 shows the release of luciferase from infected cells treated with different PRBs. The results indicate that PRB-1, 2, 6, 7 and 14 release significantly higher luciferases compared to PRB-3, 5, 9, 10, 11, 12, and 16.

Example  4

Affinity test of fatty animals with metabolic syndrome (block 4 in FIG. 2)

background

Metabolic syndrome animal models using C57BL / 6J-rich fat mice measured the release of luciferase from adipocytes of these animals, and compared with 97% DHA and the antidiabetic compound rosiglitazone, PRB-2 against PPARγ. Was used to test the affinity of 5, and 8. The animals (n = 8 in each group) were given a high fat diet (60% of total calories from fat, the same diet as in block 5) for 8 weeks. Thereafter, 1.5% fat-containing PRB was administered in one serving for two weeks. The rosiglitazone group was administered in an amount of 100 mg / KG diet. The control group continued to receive only a high fat or standard diet. 9 shows the study design.

Way

After killing, the adipose tissue was detached from other structures and cut into millimeter-sized pieces. Adipose tissue is washed with 0.9% NaCl and 5 mL of Krebs-Ringer solution containing hepse, bovine serum albumin free of fatty acids, 20OnM adenosine, 2 nM glucose and 260 U / mL collagenase Digested in a shaking water bath at 37 ° C. for 1 h 30 min. After digestion of collagenase, adipocytes were isolated from tissue fragments by filtration. The cells were then washed with Hepose, bovine serum albumin free of fatty acids, 20OnM adenosine, and Krebs-Ringer solution containing 2 nM glucose and for up to 30 minutes until electroporation. It was in a shaking water bath at 37 ° C.

Isolated initial adipocytes were infected by electroporation to measure specific PPARγ response element (PPRE) activity. In this case a plasmid encoding firefly luciferase cDNA was inserted from the acyl-CoA-oxidase gene under the control of PPRE. The cells were also infected with plasmids containing the cDNA of Renilla luciferase regulated by a systemically active promoter. The PPRE-induced firefly luciferase activity was normalized according to lenilla luciferase and thus corrected for potential differences in the amount of infected cells. Dual-Luciferase® Reporter assay system (Promega, USA) was used for luciferase signal measurement. Collected epididimal adipose tissue was sufficient to isolate adipocytes for two runs. Each group was sacrificed on two independent days and four independent infections were done for each diet group.

result

During the first eight weeks of the HF diet (33,7% o fat, w / w), there was a gradual weight gain compared to the control rats on the chow diet (4,5% w / w). there was. During the last two weeks of the experimental diet, animals fed a high fat diet and animals fed a high fat diet with rosiglitazone continued to gain weight at about the same rate as before. The weight loss of PRB-8 and PRB-5 with enough HF diet. However, in the case of PRB-2 and DHA (5% w / w) the diet completely stopped gaining weight and even showed weight loss (FIG. 10). Food consumption was occasionally recorded (4x). There was no difference between the HF group and the control group.

When rosiglitazone and high fat were mixed, PPARγ endogenous was nearly 2 times higher than other diet groups (FIG. 11). Furthermore. These adipocytes are more sensitive, for example, to additional stimulation with 5 uM rosiglitazone (5, 12 fold stimulation) compared to the HF diet itself (1, 5 fold stimulation). This rosiglitazone sensitivity effect was also seen in the PRB-2 and PRB-5 diet groups (2 and 6 fold stimulation).

The data in this study demonstrate the activity on nuclear PPAR receptors and show the most significant effect on body weight, especially in the group receiving PRB-2. Even in animals fed PRB-5 and PRB-8, weight gain did not occur as in the high-fat diet group. Interestingly, animals that received rosiglitazone gained weight to the same extent as animals that received only a high fat diet. This clearly demonstrates the negative effect of risking weight gain even if insulin resistance is reduced when only PPARγ ligands such as glilitazone are given. However, in view of PPARγ activity measured by luciferase activity in this experiment, rosiglitazone has a higher value than any other PRB. Within the PRB group, PRB-2 and PRB-5 had higher values than those with only PRB-8 and DHA (FIG. 12).

Example  5

Pharmacodynamic Effects of DHA Derivatives in Metabolic Syndrome Animal Models (Block 5 of FIG. 2)

background

Metabolic syndrome animal models using C57BL / 6J-rich lipid mice were used to examine the effects of typical experiments on metabolic syndrome and common pathological anatomical features. Animals given a high fat diet containing about 60% fat became obese and had livers with high insulin plasma levels, pathological glucose tolerance tests, increased plasma triglycerides and non-esterified fatty acid development.

Example 5a

Effect of DHA Derivatives in Adipose Rats During 4 Month Diet

Way

All experiments were either subfamily C57BL / N (provided by Charles River, Germany, n = 160, experiment AC, see below) or subfamily C57BL / 6J (provided by the Jackson laboratory, Bar Harbor). ), Main (ME), USA, n = 32, experiment D), C57BL / 6 mice males. Due to selection, the total number of animals used (n = 170 and 36, respectively) was large. In the latter case, animals were raised for several generations (<20) in the Institute of Physiology. At the beginning of the treatment animals were 14 weeks old and weighed between 23.6 and 27.1 g. One week before the start of the study, animals were divided by weight and similar mean weights were divided into subgroups (n = 8). This method allowed about 5-10% of the lowest and highest weight animals to be culled. Animals excluded from this study were sacrificed by cervical dislocation. Complete health checks of rats were performed by donor Charles River and serological tests were performed by ANLAB (Prague, Czech Republic) earlier in the study. In addition, regular health checks were performed at animal farms every three months using surveillance rats and serological tests (ANLAB). In all tests the animals were free of specific pathogens.

Diet

Animals were given three types of experimental diets.

(i) ssniff RM-H sold by the SSNIFF Spezialdieten Gmbh of Soest, Germany, which includes proteins, fats and carbohydrates that form 33%, 9 and 58% energy, respectively; http://ssniff.de Reference)

(ii) a high fat diet (cHF diet) and fatty acid composition prepared in the laboratory with proteins, fats and carbohydrates that form 15, 59 and 26% energy, respectively (most of the fat is obtained from corn oil; see Ruzickova 2004).

 (iii) cHF diet in which 0.15, 0.5 and 1.5% of fat (particularly corn oil components) is replaced with DHA or various PRB compounds such as PRBl, PRB2, PRB5, PRB7 and PRB8, respectively. All compounds were in ethyl ester form and provided sealed by Pronova Biocare a.s. The chemical composition of the PRB compound was unknown to the laboratory conducting the experiment (biochemistry laboratory, Academy of Sciences Prague, Czech Republic).

After arrival, the PRB compound was stored in the refrigerator in the original container. The vessel was opened just before preparing the experimental diet. The diet was stored in plastic bags injected with nitrogen and stored at −70 ° C. in small amounts sufficient to feed the animals for a week. Fresh doses were injected every two days or daily.

Overview of this study

This study is based on four independent experiments. In each experiment, PRB compounds (or DHA, respectively) mixed in the cHF diet at three concentrations (fat content of 0.15, 0.5 and 1.5%) were tested. In each experiment, mice fed only the cHF diet were included as subgroups and served as controls. Animals (n = 8-13) were housed in cages of four, if randomly assigned a different test diet, and provided until the basic chow diet was three months old. Two months after the new diet (5 months), the animals fasted overnight and the Glucose Tolerance Test (GTT) test was performed in the morning.

Research parameters

The parameters of this study were: weight gain (grams), area under the curve graph of peritoneal glucose tolerance test (mMol x 180 min) (AUC), plasma insulin (ng / ml), serum triglycerides ( TAGs ₅ mmol / l), and non-esterified fatty acids (NEFA ₅ mmol / l).

FIG. 13 shows typical glucose clearance curves before and after administration of a compound with insulin resistance in animals with insulin resistance. The reduction in the area under the curve graph means that blood glucose due to reduced insulin resistance was more effectively removed.

result

The results are shown in the following Tables 2, 3 and 4 (* = significant difference compared to cHF diet (P <0.05)).

Table 2 shows the effects in animals administered a 1.5% concentration of PRB test compound compared to animals administered a high fat diet (cHF) or standard chow diet (STD) mixed with 97% DHA. Body weight gain was significantly reduced in animals administered PRB-2 compared to animals fed a high fat diet (cHF). Food intake was somewhat lower in this group. Significant decreases in AUC in glucose tolerance tests were seen in the same group and in animals that received PRB-I. Plasma insulin was shown to be somewhat effective in PRB-I and PRB-5 animals, but the PRB-2 group was significantly lower than the cHF control group. The PRB-2 group also showed the largest reduction in triglycerides (TAGs) and non-esterified fatty acids (NEFA).

Table 3 shows the effect in animals administered with a lower concentration of 0.5% PRB test compound compared to animals administered a high fat diet (cHF) or a standard chow diet (STD) mixed with 97% DHA. Body weight gain was somewhat lower in animals administered PRB-2 and PRB-5. In the glucose tolerance test as well as plasma insulin, AUC was significantly reduced only in the PRB-2 group.

Table 4 shows the results obtained from the lowest PRB concentration of 0.15%. The difference here was small. Only AUC in the PRB-2 group was significantly lower, while weight gain was somewhat lower in the PRB-I and PRB-2 groups. Plasma insulin was low in PRB-1, 2 and 7.

TABLE 2 1. Effect of PRB derivative after 4 months at 5% concentration

TABLE 3 Effect of PRB derivatives after 4 months administration at 0.5% concentration

TABLE 4 Effect of PRB Derivatives after Administration for 4 Months at 0.15% Concentration

In conclusion, tests of PBR-1, 2, 5 and 7 for 4 months in fat-rich animals with insulin resistance and metabolic syndrome, especially in DHA derivative PBR-2, reduced triglycerides and non-esterified fatty acids. In addition, insulin resistance and metabolic syndrome, such as insulin resistance and weight loss, reduced AUC in peritoneal glucose tolerance tests, and low insulin / plasma levels, showed positive and unexpected effects in PRB tested animals. Effects were observed in the 1.5% group as well as the 0.5% group. Some effects even occurred at 0.15% of the lowest concentration.

Testing of the PRB-8 compound was started later, so data were obtained from only 2 months in the 3 groups (1.5%, 0.5% and 0.15%) that had been administered. In the group given 1.5%, the body weight of the control group was 28.0 ± 0.7 grams compared to 29.6 ± 0.9 grams and AUC was 1031 ± 104 compared to 1074 ± 91. This difference is small but the trend was interesting. There was no difference between intraperitoneal and control groups at 0.5% and 0.15% doses. Data on PRB-8 data from two months of dosing animals show trends in weight loss and AUC.

Example 5b

Effect of DHA Derivatives on Metabolic Syndrome and Insulin Resistance

Way

In another example, PRB-2, PRB-5 and PRB-7 were tested in the same animal breeding method. In this experiment, animals were initially given a high fat diet (in the same way as in Example 5a) for 8 weeks and were given PRB after the development of insulin resistance and metabolic syndrome. Initial administration was 15% of fat with PRB but no resistance. Two weeks later the animals received 1.5% PRB-2, 5% and 1.5% PRB-5 and 1.5% and 0.5% PRB-7.

result

Weight loss was very well seen in animals administered PRB-2. Animals receiving PRB-5 showed some weight loss but at a 5% higher dose. Triglycerides were decreased in all derivatives tested compared to the animal control group on a mixed high fat diet. The decrease in non-esterified fatty acids was most pronounced in PRB-2 and PRB-5 and their doses were different (see FIG. 14).

Blood cholesterol was decreased in animals administered PRB-2 and PRB-5. Blood glucose was not affected by the fact that animals were pre-diabetic with normal glucose because of high insulin production. More importantly, however, PRB-2 showed a significant decrease at much lower concentrations compared to DHA-derived PRB-5, which showed the second largest reduction in plasma insulin. Even PRB-7 had some effect on insulin concentrations (see Figure 15).

PRB-2 showed a significant decrease in AUC blood glucose at each point in the curve when compared to baseline. This means that blood glucose was more effectively removed after 1.5% of PRB-2 administration. PBR-5 and PBR-7 showed some effects but the degree was not the same (see FIG. 16).

This effect is very unexpected and is highly related to the positive effects of metabolic syndrome and type 2 diabetes. These patients are almost overweight or obese and the drug's weight loss effect is very positive. The most useful treatment used today for the treatment of type 2 diabetes is thiazolidinedione, which is a potential PPARγ ligand and thus reduces insulin resistance but results in very negative weight gain in patients with this disease (Yki-Jarvinen 2004). .

Reduction of serum triglycerides is another very important effect that demonstrates this experiment. Patients with metabolic syndrome and type 2 diabetes generally have high triglycerides. The triglyceride reducing effect of DHA derivatives is a positive finding and PRB-2 has the most potential effect at the lowest dose. The effects in plasma insulin and glucose tolerance tests are very promising and very unexpected. The effects obtained together with the DHA derivative, especially PRB-2, are very promising to form a good basis for the development of antidiabetic drugs.

Example 5c

Test of DHA Derivatives in Liver Fat

Way

Tissue samples obtained from animals receiving DHA derivatives in this Example were analyzed histologically. After paraffination, tissue samples from liver, adipose tissue, skeletal muscle, pancreas and kidney were stained with eosin-hematoxylin.

result

There were no pathological findings in the tested tissues except for tissue samples from the liver. Control animals on a high fat diet developed fatty liver (liver steatosis). Fat droplets in the liver are easily distinguishable from normal liver cells. Animals administered PRB-I, 5 and 7 showed low fatty liver. However, animals receiving 1.5% PRB-2 had complete normal hepatocytes with no signs of steatosis. This is a very important finding and is very positive for the treatment of patients with insulin resistance, obesity and type 2 diabetes. Hepatic steatosis is commonly found in these patients and is associated with the overloading of fatty acids and triglycerides, which are biological characteristics in the development of insulin resistance and metabolic syndrome. DHA derivatives reduce hepatic steatosis and PRB-2 is the most effective compound showing this effect.

Discussion and conclusion

The present invention clearly demonstrates a group of new compounds that activate nuclear receptors, particularly PPARα and PPARγ, and provide therapeutic effects in the treatment of insulin resistance or metabolic syndrome, type 2 diabetes cardiovascular disease and other atherosclerosis related diseases. .

The elements of this group are DHA derivatives with a different kind of side branch at the alpha position of the molecule. Many alpha-substituted DHA derivatives have been tested and compared with controls as well as pure DHA and EPA. Several compounds tested have demonstrated many interesting biological effects associated with potential antidiabetic drugs.

Alpha-ethyl DHA ethyl ester (PRB-2) is used to show the effects on diseases that primarily cause pathophysiological diseases such as insulin resistance and metabolic syndrome, type 2 diabetes cardiovascular disease and other atherosclerosis related diseases. All tests showed significant effects. Alpha-ethyl DHA ethylester was abundant in liver tissue obtained from animals administered with the other DHA derivatives tested (block 1), indicating that the compound is not used for the synthesis of triglycerides, eicosanoids or other metabolic intermediates. Indirectly this means that alpha-ethyl DHA can be a ligand for nuclear receptors such as PPARs.

In testing affinity for PPARα and PPARγ using computer-based docking techniques, many DHA derivatives showed affinity for both receptors, particularly the most important nucleus associated with the activity of genes involved in blood glucose metabolism. It showed the highest affinity of the receptor, PPARγ. In particular alpha-diethyl DHA (PBR-8) as well as alpha-ethyl DHA (PRB-2) had excellent affinity for nuclear receptors. Compared with alpha-diethyl DHA (PBR-8), alpha-ethyl DHA has two stereoisomers, type r and s. Using the docking technique, both stereoisomers showed the same affinity for PPARα and PPARγ, which means that r-type s has no advantage over racemic type. In fact, racemic forms may have advantages over each of the stereoisomers. When testing the affinity of infected cells carrying nuclear receptors and accompanying DNA response elements, some PRBs showed good affinity and affinity was measured by the release of luciferase. Alpha-ethyl DHA (PRB-2) with PRB-6, 7 and 14 demonstrated the best effect.

Five of the DHA derivatives were extensively tested in the C57BL / 6 rat model, which showed insulin resistance and metabolic syndrome when fed a high fat diet. PRB-1, 5 and 7 were tested in two and alpha-diethyl DHA (PBR-8) was tested in one, while alpha-ethyl DHA (PRB-2) was tested in three independent experiments. All derivatives showed significant biological effects. However, alpha-ethyl DHA (PBR-2) showed the most significant effect on the continuous reduction of AUC and plasma insulin in body weight and peritoneal glucose tolerance tests, as well as triglycerides and non-esterified fatty acids. The effect was obtained at 1.5% and 0.5% administration. The 0.15% administration tested at the lowest concentration did not perform convincingly. 1.5% dose of alpha-ethyl DHA (PRB-2) demonstrated normalization of fatty liver, an important pathological condition in patients and animals with insulin resistance and metabolic syndrome.

Compared to pure DHA, alpha-ethyl DHA (PRB-2) appears to be 10-30 times more potent than DHA. All of these findings and potencies compared to the parent molecule DHA were unexpected and very unexpected.

Because alpha-ethyl DHA (PRB-2) religates the nuclear receptors PPARα and PPARγ simultaneously, the compounds not only have therapeutically effective effects in glucose and fat metabolism, especially in insulin resistance, metabolic syndrome and type 2 diabetes. It has general anti-inflammatory effect and weight loss effect. Alpha-ethyl DHA (PRB-2), either directly or through positive adjustments to risk factors, has a cardiovascular preventive effect as well as a prophylactic effect of cardiovascular diseases such as myocardial infarction and stroke.

Drugs that act as PPAR ligands are already on the market, but even though these compounds have a positive effect on glucose metabolism, they are problematic due to side effects such as high triglycerides, weight gain and edema. The alpha-substituted DHA derivatives shown in this application have a combination of PPARα and PPARγ that may be appropriate and useful for insulin resistance, metabolic syndrome and type 2 diabetes. Moreover, these combined actions also have important effects on blood fat, inflammation, atherosclerosis and cardiovascular disease.

The invention is not limited to the examples and examples shown herein.

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Claims (130)

A compound of formula (I) or a pharmaceutically acceptable salt, solvate, complex, or prodrug thereof:  Formula (I)]

R ₁ and R ₂ are the same or different from each other, a hydrogen atom, an alkyl group, a halogen atom, an alkoxy group, an acyloxy group, an acyl group, an alkenyl group, an alkynyl group, an aryl group, an alkylthio group, an alkoxycarbonyl group, an alkylsulfinyl group, An alkylsulfonyl group, an amino group, and an alkylamino group; And X is a carboxylic acid group or a carboxylate group; The compound of formula (I) is not alpha-methyl DHA, alpha-methyl DHA methyl ester, alpha-methyl DHA ethyl ester or alpha-hydroxy DHA ethyl ester, and R ₁ and R ₂ are not simultaneously hydrogen atoms. The compound of formula (I) according to claim 1, wherein the alkyl group is selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-butyl, sec-butyl and n-hexyl Pharmaceutically acceptable salts, solvates, complexes, or prodrugs. The compound of formula (I) according to claim 1, wherein the alkenyl group is selected from the group consisting of allyl, 2-butenyl, and 3-hexenyl Pharmaceutically acceptable salts, solvates, complexes, or prodrugs. The compound of formula (I) or a pharmaceutically acceptable thereof according to claim 1, wherein the alkynyl group is selected from the group consisting of propargyl, 2-butynyl, and 3-hexynyl. Possible salts, solvates, complexes, or prodrugs. The compound of formula (I) or a pharmaceutically acceptable salt, solvate, complex, or prodrug thereof according to claim 1, wherein the halogen atom is a fluorine atom. The compound of claim 1, wherein the alkoxy group is selected from the group consisting of methoxy, ethoxy, propoxy, isopropoxy, sec-butoxy, phenoxy, benzyloxy, OCH ₂ CF ₃ and OCH ₂ CH ₂ OCH ₃ A compound of formula (I) or a pharmaceutically acceptable salt, solvate, complex, or prodrug thereof. The compound of formula (I) or a pharmaceutically acceptable salt, solvate, complex, or prodrug thereof according to claim 1, wherein the aryl group is a phenyl group. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, wherein the alkoxycarbonyl group is selected from the group consisting of methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl and butoxycarbonyl. Solvates, complexes, or prodrugs. According to claim 1, The alkylsulfinyl group is selected from the group consisting of methanesulfinyl, ethanesulfinyl and isopropanesulfinyl, is a compound of formula (I) or pharmaceutically Acceptable salts, solvates, complexes, or prodrugs. The compound of formula (I) or a pharmaceutically acceptable salt, solvate thereof according to claim 1, wherein the alkylsulfonyl group is selected from the group consisting of methanesulfonyl, ethanesulfonyl and isopropanesulfonyl , Complexes, or prodrugs. The compound of formula (I) or a pharmaceutically acceptable salt, solvate thereof according to claim 1, wherein the alkylamino group is selected from the group consisting of methylamino, dimethylamino, ethylamino, and diethylamino. , Complexes, or prodrugs. The carboxylate group of claim 1, wherein the carboxylate group is ethyl carboxylate, methyl carboxylate, n-propyl carboxylate, isopropyl carboxylate, n-butyl carboxylate, sec-butyl carboxylate, and n A compound of formula (I) or a pharmaceutically acceptable salt, solvate, complex, or prodrug thereof, selected from the group consisting of hexyl carboxylates. The compound of formula (I) or a pharmaceutically acceptable salt, solvate thereof according to claim 1, wherein R ₁ is a hydrogen atom, R ₂ is an alkyl group or an alkoxy group, and X is a carboxylate group. ), Complexes, or prodrugs. The compound of claim 13, wherein R ₁ is a hydrogen atom, R ₂ is an ethyl group, and X is a chemical name of ethyl (all-Z) -2-ethyldocosa-4,7,10,13,16,19- A compound of formula (I) or a pharmaceutically acceptable salt, solvate, complex, or prodrug thereof, which is an ethyl carboxylate group that is hexaenoate. The compound of claim 13, wherein R ₁ is a hydrogen atom, R ₂ is an ethyl group, and X has a chemical name of methyl (all-Z) -2-ethyldocosa-4,7,10,13,16,19- A compound of formula (I) or a pharmaceutically acceptable salt, solvate, complex, or prodrug thereof, which is a methyl carboxylate group that is hexaenoate. The compound of formula (I) or a pharmaceutically acceptable salt, solvate, complex thereof, according to claim 1, wherein R ₁ and R ₂ are not identical to each other and are present as S optical isomers Prodrugs. The compound of formula (I) or a pharmaceutically acceptable salt, solvate, complex thereof, according to claim 1, wherein R ₁ and R ₂ are not identical to each other and are present as R optical isomers. Prodrugs. The compound of formula (I) or a pharmaceutically acceptable salt, solvate, complex, or precursor thereof according to claim 1, wherein R ₁ and R ₂ are not identical to each other and are present as racemates. Drug (prodrug). The compound of formula (I) or a pharmaceutically acceptable salt, solvate thereof, according to claim 1, wherein R ₁ and R ₂ are not identical to each other and are present in a mixture of the R optical isomer and the S optical isomer. Complex, or prodrug. The compound of formula (I) or a pharmaceutically acceptable salt, solvate, or complex thereof according to claim 1, wherein R ₁ and R ₂ are both ethyl groups and X is a carboxylic acid group or a carboxylate group. ), Or prodrugs. The compound of formula (I) or a pharmaceutical thereof, according to claim 1, wherein X is COO ^- Z ⁺ , wherein Z ^{+ is in the} form of a salt which ^is Li ⁺ , Na ⁺ , K ⁺ , NH ₄ ⁺ , or substituted NH ₄ ⁺ Optionally acceptable salts, solvates, complexes, or prodrugs. The compound of formula (I) or a pharmaceutically acceptable salt, solvate, complex, or prodrug thereof according to claim 1, in the form of a salt having the formula

Wherein Z ²⁺ is Mg ²⁺ or Ca ²⁺ . The compound of formula (I) or a pharmaceutically acceptable salt, solvate, complex, or prodrug thereof according to claim 1, wherein X is a carboxylic acid derivative in the form of a trisleyide or phospholipid. ). A compound of formula (I) or a pharmaceutically acceptable salt, solvate, complex, or prodrug thereof:  Formula (I)]

R ₁ and R ₂ are the same or different from each other, a hydrogen atom, a hydroxy group, an alkyl group, a halogen atom, an alkoxy group, an acyloxy group, an acyl group, an alkenyl group, an alkynyl group, an aryl group, an alkylthio group, an alkoxycarbonyl group, an alkylsulfide It is selected from the group consisting of a silyl group, an alkylsulfonyl group, an amino group, and an alkylamino group; And X is a carboxylic acid group; R ₁ and R ₂ are not hydrogen atoms at the same time. The chemical formula of claim 24, wherein R ₁ is a hydrogen atom and R ₂ is an ethyl group having a chemical name (all-Z) -2-ethyldocosa-4,7,10,13,16,19-hexaenoic acid. A compound of (I) or a pharmaceutically acceptable salt, solvate, complex, or prodrug thereof. The compound of formula (I) or a pharmaceutically acceptable salt, solvate, complex, or prodrug thereof according to claim 25, wherein the carboxylic acid group is in the form of triglyceride or phospholipid. A compound of formula (I)  Formula (I)]

Wherein R ₁ and R ₂ are the same or different from each other, a hydrogen atom, a hydroxyl group, an alkyl group, a halogen atom, an alkoxy group, an acyloxy group, an acyl group, an alkenyl group, an alkynyl group, an aryl group, an alkylthio group, an alkoxycarbonyl group, An alkylsulfinyl group, an alkylsulfonyl group, an amino group, and an alkylamino group; And X is a carboxylic acid group in which carboxylic acid is present as its diglyceride derivative; R ₁ and R ₂ are not hydrogen atoms at the same time. 28. The compound of claim 27, wherein said alkyl group consists of methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, and n-hexyl. A process for preparing the compound of claim 1 comprising the following steps: a) reacting the DHA ethyl ester with a strong non-nucleophilic base, and b) reacting the solution obtained in step a) with an alkylation reagent, and c) extracting the product obtained from the solution obtained in step b) with a solvent, and d) optionally converting said ester into a carboxylic acid or carboxylic acid derivative. The method of claim 29, wherein said alkylating reagent is selected from alkyliodides. The method of claim 29, wherein the DHA ethyl ester is prepared from a source that is a plant, microorganism, animal, or a combination thereof. The method of claim 29, wherein said DHA ethyl ester is prepared from marine oil. 33. The method of claim 32, wherein the marine oil is fish oil. The method of claim 29, wherein said strong non-nucleophilic base is selected from lithium diisopropylamide, potassium hexamethyldisilazane, and sodium hexamethyldisilazane. A process for preparing a compound according to formula (I) or a pharmaceutically acceptable salt, solvate, complex, or prodrug thereof, comprising the steps of:

(I) a) reacting the DHA ethyl ester with a strong non-nucleophilic base, and b) reacting the solution obtained in step a) with ethyl iodide, and c) extracting the product obtained from the solution obtained in step b) with a solvent. Wherein X is carboxylic acid or a derivative thereof, carboxylate, carboxylic anhydride or carboxamide. 36. The method of claim 35, wherein the ester obtained in step c) is converted to a carboxylic acid derivative. 36. The method of claim 35, wherein said DHA ethyl ester is prepared from a plant, microorganism, animal source, or a combination thereof. 36. The process of claim 35, wherein said DHA ethyl ester is marine oil. The method of claim 38, wherein the marine oil is fish oil. 36. The process of claim 35, wherein said strong non-nucleophilic base is selected from lithium diisopropylamide, potassium hexamethyldisilazane, and sodium hexamethyldisilazane. . A medicament according to claim 1 for the treatment or prevention of peripheral insulin resistance, diabetes symptoms, obesity or overweight symptoms, and an inflammatory disease or condition. 42. The agent according to claim 41, wherein the diabetes symptom is type II diabetes. A medicament according to claim 1 for the treatment or prevention of dyslipidemia. 44. The agent of claim 43, wherein the dyslipidemia is a hyperlipidemic condition. 44. The agent of claim 43, wherein the dyslipidemia comprises at least one of increased triglyceride levels or non-HDL cholesterol (LDL cholesterol and VLDL cholesterol levels). A medicament according to claim 1 for reducing at least one of insulin, blood sugar, or plasma triglyceride levels. A medicament according to claim 1 for the treatment of hypertriglyceridemia. A medicament according to claim 1 for reducing weight or preventing weight gain. A medicament according to claim 1 for activating or mediating at least one human peroxysome proliferator-activated receptor (PPAR) isoform. 50. A pharmaceutical composition comprising at least one medicament according to any one of claims 41 to 49. 51. The method of claim 50, wherein the at least one agent is ethyl (all-Z) -2-ethyldocosa-4,7,10,13,16,19-hexaenoate, methyl (all-Z) -2. Ethyl docosa-4,7,10,13,16,19-hexanoate, and (all-Z) -2-ethyl docosa-4,7,10,13,16,19-hexaenoic acid Pharmaceutical composition selected from. Lipid composition comprising at least one compound according to claim 1. The compound of claim 52, wherein the at least one compound is ethyl (all-Z) -2-ethyldocosa-4,7,10,13,16,19-hexaenoate, methyl (all-Z) -2. Ethyl docosa-4,7,10,13,16,19-hexanoate, and (all-Z) -2-ethyl docosa-4,7,10,13,16,19-hexaenoic acid A lipid composition selected from. (4Z, 7Z, 10Z, 13Z, 16Z, 19Z) -2-ethyldocosa-4,7,10,13,16,19-hexaenoic acid compound according to formula (I) Formula (I)]

55. The compound of claim 54, wherein the compound is present as an S optical isomer. 55. The compound of claim 54, wherein the compound is present as the R optical isomer. 55. The compound of claim 54, wherein the compound is in a mixture of R and S optical isomers. The compound of claim 57, wherein the compound is a racemate. A composition comprising at least one compound according to claim 54. 55. A process for the preparation of the compound of claim 54 comprising the following steps: a) reacting a solution comprising a DHA ester with a strong non-nucleophilic base to provide an ester enolate, b) reacting the enolate obtained from step a) with an electrophilic ethyl reagent, and c) adding a base in aqueous solution to hydrate the product obtained in step b) in a solvent. 61. The method of claim 60, wherein said DHA ester is prepared from a plant, microorganism, animal source, or a combination thereof. The method of claim 61, wherein the DHA ester is prepared from marine oil. 63. The method of claim 62, wherein said marine oil is fish oil. 61. The method of claim 60, wherein said strong non-nucleophilic base is selected from lithium diisopropylamide, potassium hexamethyldisilazane, and sodium hexamethyldisilazane. 65. The method of claim 64, wherein said strong non-nucleophilic base is lithium diisopropylamide. 61. The method of claim 60, wherein said solvent is selected from ethanol or methanol. 61. The method of claim 60, wherein said electrophilic ethyl reagent is ethyl iodide. 55. A medicament according to claim 54 for the treatment or prevention of diabetes symptoms. The agent of claim 68, wherein the diabetic symptom is type II diabetes. The medicament according to claim 54 for the reduction of at least one of insulin, blood glucose, or plasma triglyceride levels. 55. A medicament according to claim 54 for the treatment or prevention of dyslipidemia. 72. The agent of claim 71, wherein the dyslipidemia is a hyperlipidemic condition. 73. The medicament of claim 72, wherein the hyperlipidemia is hypertriglyceridemia. 55. A medicament according to claim 54 for the reduction of cholesterol in the blood. 55. A medicament according to claim 54 for the enhancement of HDL cholesterol levels in plasma. 55. A medicament according to claim 54 for the reduction of body weight or the prevention of weight gain. 55. A medicament according to claim 54 for the treatment or prevention of peripheral insulin resistance. 78. A pharmaceutical composition comprising a medicament according to any one of claims 68 to 77. A pharmaceutical composition according to claim 78 of a formulation for oral administration. A pharmaceutical composition according to claim 78 in a capsule or sachet formulation. A pharmaceutical composition according to claim 78 in the form of a solid. A pharmaceutical composition according to claim 78 of the intravenous, subcutaneous, or intramuscular formulation. A pharmaceutical composition according to claim 78 of the formulation for providing a daily dosage of 10 mg to 10 g. The pharmaceutical composition of claim 83 of the formulation for providing a daily dosage of 100 mg to 1 g. A lipid composition comprising the compound of claim 54. 86. The lipid composition of claim 85, wherein said compound of formula (I) is present at a concentration of at least 60% by weight of the total lipid composition. 87. The lipid composition of claim 86, wherein the compound of formula (I) is present at a concentration of at least 90% by weight of the total lipid composition. 86. The lipid composition of claim 85, further comprising an antioxidant. 89. The lipid composition of claim 88, wherein said antioxidant is tocopherol. Compounds according to formula (I) or pharmaceutically acceptable salts, solvates, complexes, or prodrugs thereof:

(I) Wherein X is carboxylic acid or a derivative thereof, carboxylate, carboxylic anhydride, or carboxamide. 91. The compound according to formula (I) or a pharmaceutically acceptable salt, solvate, complex, or precursor thereof according to claim 90, wherein X is a diglyceride, triglyceride, or carboxylic acid derivative in phospholipid form. Drug (prodrug). 92. The compound according to formula (II), or a pharmaceutically acceptable salt, solvate, complex, or prodrug thereof, according to claim 91, wherein X is carboxylic acid in triglyceride form.

(II) 91. A compound according to formula (II) or a pharmaceutically acceptable salt thereof, according to claim 90, wherein X is COO ^- Z ⁺ and Z ⁺ is in the form of a salt which is Li ⁺ , K ⁺ , NH 4 ⁺ , or substituted NH 4 ⁺ . Solvates, complexes, or prodrugs. 91. The compound of claim 90, or a pharmaceutically acceptable salt, solvate, complex, or prodrug thereof, in the form of a salt according to the formula:

Wherein Z ²⁺ is Mg ²⁺ or Ca ²⁺ . 91. The method of claim 90, wherein the carboxylate group is ethyl carboxylate, methyl carboxylate, n-propyl carboxylate, isopropyl carboxylate, n-butyl carboxylate, sec-butyl carboxylate, and n A compound according to formula (I) or a pharmaceutically acceptable salt, solvate, complex, or prodrug thereof, wherein the compound is selected from hexyl carboxylate. 91. The method of claim 90, wherein the carboxamide group is a primary carboxamide, N-methyl carboxamide, N, N-dimethyl carboxamide, N-ethyl carboxamide, and N, N-diethyl carboxamide A compound according to formula (I) selected from pharmaceutically acceptable salts, solvates, complexes, or prodrugs thereof. 98. The compound according to formula (I) or a pharmaceutically acceptable salt, solvate, complex, or precursor thereof according to claim 95, wherein said carboxylate group is methyl carboxylate or ethyl carboxylate Drug (prodrug). 91. The compound of claim 90, or a pharmaceutically acceptable salt thereof, which is ethyl (all-Z) -2-ethyldocosa-4,7,10,13,16,19-hexaenoate according to the formula: Solvates, complexes, or prodrugs.

91. The compound according to formula (I), or a pharmaceutically acceptable salt, solvate, complex, or prodrug thereof, according to claim 90, present as S optical isomer. 91. A compound according to formula (I) or a pharmaceutically acceptable salt, solvate, complex, or prodrug thereof according to claim 90, which is present as R optical isomer. 91. The compound according to formula (I) or a pharmaceutically acceptable salt, solvate, complex, or prodrug thereof according to claim 90, which is present in a mixture of R and S optical isomers. 102. The compound of formula (I) or a pharmaceutically acceptable salt, solvate, complex, or prodrug thereof, according to claim 101, wherein said mixture is racemic. A composition comprising at least one compound according to claim 90. 107. The composition of claim 103, further comprising a compound of formula

Peripheral insulin resistance; Diabetes symptoms; Obesity or overweight symptoms; And an inflammatory disease or condition; a medicament according to claim 90 for the treatment or prevention of a disease or condition selected from. 107. The agent of claim 105, wherein the diabetic condition is type II diabetes. 90. A medicament according to claim 90 for the treatment or prevention of dyslipidemia. 107. The agent of claim 107, wherein the dyslipidemia is a hyperlipidemic condition. 109. The medicament of claim 108, wherein the dyslipidemia comprises at least one of increased triglyceride levels or non-HDL cholesterol (LDL cholesterol and VLDL cholesterol levels). 91. A medicament according to claim 90 for the reduction of at least one of insulin, blood glucose or plasma triglyceride levels. 90. A medicament according to claim 90 for the reduction of body weight or the prevention of weight gain. 91. A medicament according to claim 90 for activating or mediating at least one human peroxysome proliferatively-activated receptor (PPAR) isoform. 112. A pharmaceutical composition comprising a medicament according to any one of claims 105 to 112. The pharmaceutical composition according to claim 113 of the formulation for oral administration. 113. A pharmaceutical composition according to claim 113 in a capsule or sachet formulation. The pharmaceutical composition according to claim 113 of the formulation of the solid. The pharmaceutical composition according to claim 113 of the intravenous, subcutaneous, or intramuscular formulation. The pharmaceutical composition according to claim 113, of the formulation for providing a daily dosage of 10 mg to 10 g. The pharmaceutical composition according to claim 113 of the formulation for providing a daily dosage of 100 mg to 1 g. A lipid composition comprising the compound of claim 90. 121. The lipid composition of claim 120, wherein said compound of formula (I) is present at a concentration of at least 60% by weight of the total lipid composition. 124. The lipid composition of claim 121, wherein the compound of formula (I) is present at a concentration of at least 90% by weight of the total lipid composition. 121. The lipid composition of claim 120, further comprising an antioxidant. 124. The lipid composition of claim 123, wherein said antioxidant is tocopherol. Process for preparing compound ethyl (all-Z) -2-ethyl docosa-4,7,10,13,16,19-hexaenoate comprising the following steps:

a) reacting a solution comprising a DHA ester with a strong non-nucleophilic base to provide an ester enolate, and b) reacting the enolate obtained from step a) with an electrophilic ethyl reagent, and c) extracting the product from the solution obtained in step b) with a solvent. 126. The method of claim 125, wherein said DHA ester is prepared from a plant, microorganism, animal source, or a combination thereof. 126. The process of claim 125, wherein said DHA ester is prepared from marine oil. 127. The method of claim 127, wherein said marine oil is fish oil. 126. The process of claim 125, wherein said strong non-nucleophilic base is selected from lithium diisopropylamide, potassium hexamethyldisilazane, and sodium hexamethyldisilazane. 126. The method of claim 125, wherein said electrophilic ethyl reagent is ethyl iodide.

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