JP2020079278A - Use of thia oxo compounds for lowering apo c3 - Google Patents

Abstract

To provide methods of reducing apolipoprotein C-III mRNA or protein.SOLUTION: A pharmaceutical composition for reducing apolipoprotein C-III (apoC-III) mRNA or protein in a subject in need thereof comprises a pharmaceutically effective amount of a compound of formula (I) in the figure, or a pharmaceutically acceptable salt or ester thereof. In the formula, Rand Rare independently selected from among a hydrogen atom and linear, branched and/or cyclic C-Calkyl groups, with the proviso that Rand Rare not both hydrogen.SELECTED DRAWING: None

Description

  The present disclosure provides a method of reducing apolipoprotein C-III (apo C-III) mRNA or protein in a subject in need thereof, wherein the method comprises a pharmaceutically effective amount of a compound of formula (I):

又はその薬学的に許容される塩若しくはエステルを対象に投与することを含み、
式中、R₁及びR₂は、水素原子又は直鎖、分岐、及び/若しくは環式C₁〜C₆アルキル基から独立して選択され、但しR₁及びR₂が両方とも水素であることはない、方法に関する。このような方法、化合物、及び組成物は、肝性及び/又は血漿アポC-IIIの上昇に起因するか、関連するか、又はそれにより増悪する状態、例えば高トリグリセリド血症(HTG)、高カイロミクロン血症、脂質異常症、膵炎を処置するために、並びに心血管疾患若しくは代謝異常、又はこれらの症状の1種以上の予防及び/又は処置において、有用である。

Or comprising administering to the subject a pharmaceutically acceptable salt or ester thereof,
In the formula, R ₁ and R ₂ are independently selected from a hydrogen atom or a linear, branched, and/or cyclic C ₁ -C ₆ alkyl group, provided that both R ₁ and R ₂ are hydrogen. Not about the way. Such methods, compounds, and compositions have conditions that are due to, associated with, or exacerbated by hepatic and/or elevated plasma apo C-III, such as hypertriglyceridemia (HTG), hypertension. It is useful for treating chylomicronemia, dyslipidemia, pancreatitis, and in the prevention and/or treatment of cardiovascular disease or metabolic disorders, or one or more of these conditions.

  Dietary polyunsaturated fatty acids (PUFAs), including omega-3 fatty acids, are used in normal health and chronic diseases such as regulation of plasma lipid levels, cardiovascular and immune functions, insulin action, neural development, and visual function. It has effects on a wide variety of physiological processes that are affected.

  Omega-3 fatty acids, for example, (5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaenoic acid (EPA) and (4Z,7Z,10Z,13Z,16Z,19Z) -Docosa-4,7,10,13,16,19-hexaenoic acid (DHA) regulates plasma lipid levels, cardiovascular and immune functions, insulin action, and neural development, and visual function. Omega-3 fatty acids have been shown to have beneficial effects on risk factors for cardiovascular diseases such as hypertension and hypertriglyceridemia (HTG), and on the activity of coagulation factor VII phospholipid complex.

  WO2010/128401 shows that 2-((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaenyloxy)butanoic acid favors lipid profile compared to controls. To inhibit the development of atherosclerosis in the arteries, reduce total cholesterol, and increase HDL cholesterol. These results indicate that 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid and its derivatives are associated with inflammation and hyperlipidemia. , Obesity, fatty liver disease, atherosclerosis, peripheral insulin resistance, and/or may be useful in the prevention or treatment of various conditions such as diabetic conditions. Further use of 2-((5Z,8Z,11Z,14Z,17Z)-icos-5,8,11,14,17-pentaenyloxy)butanoic acid and its derivatives for treating various diseases or conditions Are disclosed in WO 2012/059818.

  More specifically, WO2012/059818 discloses that in a subject in need thereof, elevation of apo B, primary hypercholesterolemia (heterozygous familial and non-familial), and primary dysbetalipoproteinemia (Fredrickson). A method of treating or preventing at least one disease or condition selected from type III), which method comprises administering to a subject a pharmaceutically effective amount of a compound of formula (I). However, it has already been established that the pathways associated with apo B and apo E (abnormal β lipoproteinemia) are positively affected by compounds of formula (I), although two clinical studies in different patient populations have shown that Data surprisingly revealed that an additional apolipoprotein, apo C-III, was also strongly reduced by the compound of formula (I).

  Apo C-III is a glycoprotein mainly produced by the liver, and its function is thought to include promoting the assembly and secretion of triglyceride-rich VLDL particles from hepatocytes under lipid-rich conditions. (Sundaram M et al., J Lipid Research, Vol. 51, 2010). In plasma, apo C-III is highly associated with very low density lipoprotein (VLDL), high density lipoprotein (HDL) and chylomicron. Increased levels of apo C-III induce the development of hypertriglyceridemia. The mechanism by which apoC-III expression increases plasma triglycerides is mediated in part by inhibition of lipoprotein lipase and hepatic lipase; thus delaying the catabolism of triglyceride-rich particles. Apo C-III is also believed to inhibit hepatic uptake of triglyceride-rich particles. The clinical significance of apo C-III demonstrates that carriers of rare mutations that interfere with apo C-III function have both reduced TG levels and reduced risk of coronary/ischemic cardiovascular disease Have been established (N Engl J Med. 3 Jul 2014;371(1):22-31, Loss-of-function mutations in APOC3,triglycerides,and coronary disease).

  Long chain omega-3 fatty acids, EPA and DHA, have already been validated in the treatment of HTG. Apo C-III has recently been identified as a central regulator of triglyceride levels and also as a genetically effective target for the prevention of coronary cardiovascular disease, so that various forms and compositions of omega-3. The effects of fatty acids on plasma apo C-III levels have been studied. As an example, US2014/0221486 is a subject having a baseline fasting triglyceride of about 200 mg/dl to about 499 mg/dl or having a baseline fasting triglyceride of at least about 500 mg/dl during statin therapy. Claiming a method of reducing the level of apo C-III of any of the above by administering to a subject a pharmaceutical composition comprising about 1 g to about 4 g of ethyl eicosapentaenoate per day. US2013/0177643 is a method of lowering serum or plasma apo C-III levels which comprises substantially free acid form of EPA in an amount of at least about 50% (a/a); substantially free acid form. And a pharmaceutical composition comprising DHA in an amount of at least about 15% (a/a); DPA in an amount substantially at least about 1% (a/a) in substantially free acid form, with serum or plasma apoC Claimed is a method comprising administering in an amount and for a period sufficient to reduce -III from pretreatment levels. Yet another example is a method of reducing a lipid parameter level in a subject from a reference parameter level, wherein the lipid parameter is specifically selected from the group consisting of apo C-III, and the subject is administered a composition comprising a fatty acid. Including, at least 50% by weight of the fatty acids include omega-3 fatty acids, salts, esters, or derivatives thereof, omega-3 fatty acids include eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA), and docosahexaenoic acid DHA pair. The ratio of EPA (DHA:EPA) is less than 1:10 and the ratio of DHA to DPA (DHA:DPA) is less than 2:1, the method can be found in claiming US 2014/0094520. ..

  Effective reduction of hepatic/plasma apo C-III with orally delivered omega-3/omega-3 derivatives is attractive to selected patient populations if clinically significant reductions can be achieved Various treatment options. Although the extent to which apoC-III reduction is "clinically significant" has not yet been determined, studies in subjects with loss-of-function apoC-III mutations have shown that the reduction in apoC-III is 46% lower than in non-carriers. C-III levels have been shown to be associated with a 40% lower risk of coronary cardiovascular disease (CHD) (N Engl J Med. 3 Jul 2014;371(1):22- 31, Loss-of-function mutations in APOC3, triglycerides, and coronary disease). In addition to reducing apo C-III levels, carriers also had TG levels 39% lower than non-carriers. Loss-of-function corresponds to a lifelong effect, so treatment for the purpose of reducing apo C-III over a shorter period is associated with loss-of-function mutations if a beneficial effect on CHD can be achieved. It is believed that the aim should be near (or higher) apo C-III reduction. Compounds that more strongly reduce apo C-III because the apo C-III results achieved with naturally occurring omega-3 lipids are relatively modest (see Example 26). May provide better triglyceride lowering as well as better cardioprotective effects.

  The present disclosure provides a method of reducing apolipoprotein C-III (apo C-III) mRNA or protein in a subject in need thereof, wherein the method comprises a pharmaceutically effective amount of a compound of formula (I):

又はその薬学的に許容される塩若しくはエステルを対象に投与することを含み、
式中、R₁及びR₂は、水素原子又は直鎖、分岐、及び/若しくは環式C₁〜C₂アルキル基から独立して選択され、但しR₁及びR₂が両方とも水素であることはない、方法に関する。

Or comprising administering to the subject a pharmaceutically acceptable salt or ester thereof,
Wherein R ₁ and R ₂ are independently selected from a hydrogen atom or a straight chain, branched, and/or cyclic C ₁ -C ₂ alkyl group, provided that both R ₁ and R ₂ are hydrogen. Not about the way.

  Several metabolic diseases or conditions are closely associated with an increased risk of cardiovascular events. Such diseases or conditions include type I and type II diabetes, metabolic syndrome, dyslipidemias such as hypercholesterolemia, hyperlipidemia, mixed dyslipidemia, hypertriglyceridemia, hyperchylomicronemia. And the like, as well as various familial dyslipidemias, but are not limited thereto.

  In at least one embodiment, the disease or condition is any of hypertriglyceridemia (HTG), hyperchylomicronemia, dyslipidemia, and pancreatitis, and cardiovascular disease or dysbolism, or one of these symptoms. Selected from more than one prevention and/or treatment.

  The present disclosure also includes a method of reducing apo C-III in a subject in need thereof, wherein the method comprises a pharmaceutically effective amount of 2-((5Z,8Z,11Z,14Z,17Z)-equosa-5,8. ,11,14,17-Pentaen-1-yloxy)butanoic acid:

又はその薬学的に許容される塩若しくはエステルを対象に投与することを含む。

Or administering a pharmaceutically acceptable salt or ester thereof to a subject.

FIG. 3 discloses relative hepatic apo C-III gene expression for compounds of formula (I), control and reference compounds.

  Certain aspects of the disclosure are described in more detail below. The terms and definitions used in this application, and which are apparent in the present specification, are intended to have their meaning within the present disclosure.

  The singular forms "a", "an", and "the" include plural displays unless the context indicates otherwise.

  The terms "approximately" and "about" mean approximately the same as the number or value indicated. It is to be generally understood that the terms "about" and "about" as used herein encompass a specified amount, frequency, or ±5% of a value.

  The terms “treat”, “treating”, and “treatment” include any therapeutic application that can benefit a human or non-human mammal. Both human and veterinary procedures are within the scope of this disclosure. Treatment can be responsive to an existing condition or can be prophylactic, ie, prophylactic.

  As used herein, the terms “administer”, “administration” and “administering” refer to (1) by a health advisor or a person delegated to the doctor or under the direction of the doctor. Providing, giving, dosing and/or prescribing a compound or composition according to the present disclosure, and (2) by a human patient or his or herself, or a non-human mammal, a compound or composition according to the present disclosure. Indicates that an item is taken up, ingested or consumed.

  The term "pharmaceutically effective amount" means an amount sufficient to achieve the desired pharmacological and/or therapeutic effect, i.e. an amount of a compound of the present disclosure effective for its intended purpose. Means The needs of individual subjects/patients may vary, but it is within the skill in the art to determine the optimum range for the effective amount of the compounds of the present disclosure. Generally, dosing regimens for treatment of diseases and/or conditions using the compounds disclosed herein will depend on a variety of factors such as the type of subject/patient, age, weight, sex, diet, and/or medical condition. Can be decided.

  The term "pharmaceutical composition" means a compound according to the present disclosure in any form suitable for medical use.

  The compounds of formula (I) may exist in different stereoisomeric forms, including enantiomers, diastereomers, or mixtures thereof. It will be appreciated that the present invention includes all optical isomers of the compounds of formula (I) and mixtures thereof. Thus, compounds of formula (I) that exist as diastereomers, racemates and/or enantiomers are within the scope of the present disclosure.

  The present disclosure provides a method of reducing apolipoprotein C-III (apo C-III) mRNA or protein in a subject in need thereof, wherein the method comprises a pharmaceutically effective amount of a compound of formula (I):

又はその薬学的に許容される塩若しくはエステルを対象に投与することを含み、
式中、R₁及びR₂は、水素原子又は直鎖、分岐、及び/若しくは環式C₁〜C₆アルキル基から独立して選択され、但しR₁及びR₂が両方とも水素であることはない、方法に関する。

Or comprising administering to the subject a pharmaceutically acceptable salt or ester thereof,
In the formula, R ₁ and R ₂ are independently selected from a hydrogen atom or a linear, branched, and/or cyclic C ₁ -C ₆ alkyl group, provided that both R ₁ and R ₂ are hydrogen. Not about the way.

  In at least one embodiment, the disclosure provides a pharmaceutically effective amount of a compound of formula (I) for reducing apolipoprotein C-III (apo C-III) mRNA or protein in a subject in need thereof:

又はその薬学的に許容される塩若しくはエステルの使用であって、
式中、R₁及びR₂は、水素原子又は直鎖、分岐、及び/若しくは環式C₁〜C₆アルキル基から独立して選択され、但しR₁及びR₂が両方とも水素であることはない、使用に関する。

Or the use of a pharmaceutically acceptable salt or ester thereof,
In the formula, R ₁ and R ₂ are independently selected from a hydrogen atom or a linear, branched, and/or cyclic C ₁ -C ₆ alkyl group, provided that both R ₁ and R ₂ are hydrogen. Not about the use.

In at least one embodiment, R ₁ and R ₂ are selected from hydrogen atom, methyl group, ethyl group, n-propyl group and isopropyl group.

In at least one embodiment, R ₁ and R ₂ are selected from hydrogen atom, methyl group and ethyl group.

In at least one embodiment one of R ₁ and R ₂ is a hydrogen atom and the other of R ₁ and R ₂ is selected from C ₁ -C ₃ alkyl groups. In one embodiment one of R ₁ and R ₂ is a hydrogen atom and the other of R ₁ and R ₂ is selected from a methyl group or an ethyl group.

  In at least one embodiment, the compound is present in its various stereoisomers, such as enantiomers (R or S), diastereomers, or mixtures thereof.

  In at least one embodiment the compound is present in racemic form.

  When the compound of formula (I) is a salt of a counterion having at least one asymmetric center or an ester of an alcohol having at least one asymmetric center, the compound has a plurality of stereogenic centers. You can In these situations, the compounds of this disclosure may exist as diastereomers. Thus, in at least one embodiment, the compounds of this disclosure exist as at least one diastereomer.

  In at least one embodiment, the compound of the present disclosure is 2-((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaen-1-yloxy)butanoic acid. ..

  In at least one embodiment, the compounds of the present disclosure have the formula:

で表されるそのS及び/又はR体で存在する。

Present in its S and/or R form.

  In at least one embodiment, the disease or condition is any of hypertriglyceridemia (HTG), hyperchylomicronemia, dyslipidemia, and pancreatitis, and cardiovascular disease or dysbolism, or one of these symptoms. Selected from more than one prevention and/or treatment. In one embodiment, the disease or condition is selected from hyperchylomicronemia, any of pancreatitis, and cardiovascular disease or metabolic disorders, or in the prevention and/or treatment of one or more of these conditions. .. In one embodiment, the disease or condition is selected from either hyperchylomicronemia and pancreatitis.

  Compounds of formula (I) may be prepared, for example, as described in PCT Application No. WO2010/128401, filed May 7, 2010, and based on Examples 1-23 below.

Examples 1-23 are exemplary and those skilled in the art will understand how to apply these general methods to arrive at other compounds within formula (I). Let's see The compounds of the present disclosure may be in the form of pharmaceutically acceptable salts or esters. For example, compounds of formula (I), phospholipid, glyceride, or C ₁ -C ₆ - may be in the form of an ester such as an alkyl ester. In at least one embodiment, the ester is a glyceride or C ₁ -C ₆ - is selected from alkyl esters. In at least one embodiment, the ester is triglyceride, 1,2-diglyceride, 1,3-diglyceride, 1-monoglyceride, 2-monoglyceride, methyl ester, ethyl ester, propyl ester, isopropyl ester, n-butyl ester and tert. -Selected from butyl ester. In at least one embodiment the compound of formula (I) is present as methyl ester, ethyl ester, isopropyl ester, n-butyl ester or tert-butyl ester, for example as methyl ester or ethyl ester. It has been demonstrated by in vitro digestion studies in biorelevant media that the esters represented by formula (I) (ie ethyl and butyl esters) are rapidly hydrolyzed in the digestive tract.

Suitable salts for the present disclosure include salts of NH4 ⁺ ; metal ions such as Li ⁺ , Na ⁺ , K ⁺ , Mg2 ⁺ , or Ca2 ⁺ ; tert-butylammonium, (3S,5S,7S)-adamantane. -1-ammonium, 1,3-dihydroxy-2-(hydroxymethyl)propan-2-ammonium, protonated primary amines such as protonated aminopyridine (e.g., pyridine-2-ammonium); diethylammonium, 2, Protonated secondary amines such as 3,4,5,6-pentahydroxy-N-methylhexane-1-ammonium and N-ethylnaphthalene-1-ammonium, protonated secondary amines such as 4-methylmorpholin-4-ium Protonation of tertiary amines, 2-hydroxy-N,N,N-trimethylethane-1-aminium, etc. Protonation of quaternary amines and amino((4-amino-4-carboxybutyl)amino)methaniminium, etc. Examples include, but are not limited to, a protonated heterocycle such as guanidine or 1H-imidazol-3-ium. Further examples of suitable salts include salts of diprotonated diamines such as ethane-1,2-diammonium or piperazine-1,4-diium. Other salts according to the present disclosure may include protonated chitosan.

  In at least one embodiment, the salt is selected from sodium salt, calcium salt, and choline salt.

  The present disclosure provides a method of reducing apo C-III in a subject in need thereof, comprising administering to the subject a pharmaceutically effective amount of a compound of formula (I). The subject may be a human or non-human mammal. The compounds disclosed herein may be administered as a medicament, such as in a pharmaceutical composition.

  In at least one embodiment, the present disclosure provides a baseline fasting of about 200 mg/dl to about 499 mg/dl during statin therapy by administering to a subject a pharmaceutically effective amount of a compound of formula (I). A method for reducing apo C-III levels in a subject having triglycerides. In another embodiment, the present disclosure provides the manufacture of a medicament for reducing apo C-III levels in a subject having a baseline fasting triglyceride of about 200 mg/dl to about 499 mg/dl during statin therapy, comprising: Use of a pharmaceutically effective amount of a compound of formula (I). Apo C-III levels may be reduced by at least about 20%, at least about 25%, at least about 30% or at least about 35%.

  In at least one embodiment, the present disclosure provides a subject having a baseline fasting triglyceride of about 200 mg/dl to about 499 mg/dl by administering to the subject a pharmaceutically effective amount of a compound of formula (I). A method for reducing apo C-III levels. In another embodiment, the present disclosure provides a pharmaceutically effective amount of in the manufacture of a medicament for reducing apo C-III levels in a subject having a baseline fasting triglyceride of about 200 mg/dl to about 499 mg/dl. It relates to the use of compounds of formula (I). Apo C-III levels may be reduced by at least about 20%, at least about 25%, at least about 30% or at least about 35%.

  In at least one embodiment, the present disclosure provides a subject having a baseline fasting triglyceride of greater than 500 mg/dl during statin therapy by administering to the subject a pharmaceutically effective amount of a compound of formula (I). A method for reducing apo C-III levels. In another embodiment, the present disclosure provides a pharmaceutically effective amount of a medicament for the manufacture of a medicament for reducing apo C-III levels in a subject having a baseline fasting triglyceride above 500 mg/dl during statin therapy. It relates to the use of compounds of formula (I). Apo C-III levels may be reduced by at least about 25%, at least about 30%, at least about 35% or at least about 40%.

  In at least one embodiment, the present disclosure provides that apo C-III levels in a subject having a baseline fasting triglyceride above 500 mg/dl by administering to the subject a pharmaceutically effective amount of a compound of formula (I). To reduce. In another embodiment, the present disclosure provides a pharmaceutically effective amount of formula (I) in the manufacture of a medicament for reducing apo C-III levels in a subject having a baseline fasting triglyceride above 500 mg/dl. Relates to the use of compounds. Apo C-III levels may be reduced by at least about 25%, at least about 30%, at least about 35% or at least about 40%.

  The present disclosure also provides that a subject's apoC of at least 2.5 mmol/L (about 97 mg/dl) fasting LDL-cholesterol by administering to the subject a pharmaceutically effective amount of a compound of formula (I). -It also relates to methods of reducing III levels. In another embodiment, the present disclosure provides pharmaceuticals in the manufacture of a medicament for reducing apoC-III levels in a subject having a fasting LDL-cholesterol baseline of at least 2.5 mmol/L (about 97 mg/dl). Relates to the use of a therapeutically effective amount of a compound of formula (I). Apo C-III levels may be reduced by at least about 25%, at least about 30%, at least about 35% or at least about 40%.

  In at least one embodiment, the present disclosure is directed to a method of reducing apo C-III in a subject in need thereof, which is directed to a pharmaceutically effective amount of a dyslipidemic agent, such as a statin and a compound of formula (I). To a method comprising administering to a.

  The compositions disclosed herein may comprise at least one compound of formula (I) and optionally at least one non-active pharmaceutical ingredient, ie an excipient. Inactive ingredients are those that solubilize, suspend, thicken, dilute, emulsify the active ingredient such that the active ingredient is safe, convenient, and/or otherwise acceptable for use. , Stabilized, stored, protected, colored, scented and/or adapted for applicable and effective preparation. Examples of excipients are solvents, carriers, diluents, binders, fillers, sweeteners, aromatics, pH adjusters, viscosity adjusters, antioxidants, bulking agents, water retention agents, disintegrating agents, dissolution delays. Agents, absorption enhancers, wetting agents, absorbents, lubricants, colorants, dispersants, and preservatives are included, but are not limited to. Excipients can have more than one role or function, or can be classified into more than one group; classification is merely descriptive and not intended to be limiting. In some embodiments, for example, the at least one excipient is corn starch, lactose, glucose, microcrystalline cellulose, magnesium stearate, polyvinylpyrrolidone, citric acid, tartaric acid, water, ethanol, glycerin, sorbitol, It may be chosen from fatty substances such as polyethylene glycol, propylene glycol, cetylstearyl alcohol, carboxymethylcellulose, and hard fat or suitable mixtures thereof. In some embodiments, the compositions disclosed herein comprise at least one compound of formula (I) and at least one pharmaceutically acceptable antioxidant, such as alpha-tocopherol, beta-tocopherol. , Tocopherols such as gamma-tocopherol, and delta-tocopherol, or mixtures thereof, BHA such as 2-tert-butyl-4-hydroxyanisole and 3-tert-butyl-4-hydroxyanisole, or mixtures thereof and BHT( 3,5-di-tert-butyl-4-hydroxytoluene), or a mixture thereof.

  The compositions disclosed herein may be formulated in oral dosage forms such as soft or hard capsules of tablets or gelatin. The dosage form can be any shape suitable for oral administration such as spherical, elliptical, ellipsoidal, cubical, regular and/or irregular shapes. Compounds according to the present disclosure may be formulated using conventional formulation techniques known in the art. In some embodiments, the composition can be in the form of gelatin capsules or tablets.

  A suitable daily dosage of the compound of formula (I) may range from about 5 mg to about 2 g. For example, in some embodiments, the daily dose ranges from about 50 mg to about 1 g, about 100 mg to about 1 g, about 50 mg to about 800 mg, about 100 mg to about 800 mg, about 100 mg to about 600 mg. In at least one embodiment, the daily dose is in the range of about 200 mg to about 600 mg. The compound may be administered, for example, once, twice or three times daily. In at least one embodiment, the compound of formula (I) is administered in an amount in the range of about 200 mg to about 800 mg per dose. In at least one embodiment, the compound of formula (I) is administered once per day.

  The present inventors have found that compounds of formula (I) such as 2-((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaen-1-yloxy)butanoic acid are , Has a remarkably good pharmaceutical activity. Surprisingly, the compounds of formula (I) disclosed herein exhibit improved biological activity for reducing apo C-III compared to naturally occurring omega-3 fatty acids such as EPA and DHA. Show.

  In some embodiments, for example, a compound of formula (I) may reduce median levels of apo C-III in plasma or liver by at least 25-30% relative to baseline, i.e., available Can be greatly reduced than that achieved with various EPA/DHA/DPA combinations. Compounds of formula (I) have been shown to reduce hepatic apo C-III mRNA in preclinical models (and thus probably also hepatic production/secretion), thus leading to hepatic uptake of apo B particles. It can be expected to exert a further plasma apo C-III lowering effect by the addition of lipid lowering agents such as eg statins or PCSK-9 inhibitors which reduce apo C-III via an increase in Apo C-III.

[Example]
The present disclosure can be further illustrated by the following non-limiting examples, which illustrate standard techniques known to those of skill in the art and techniques similar to those described in these examples. , May be used where appropriate. A person of ordinary skill in the art will recognize that additional embodiments will be conceivable which are consistent with the disclosure provided herein.

  Unless otherwise stated, reactions were carried out at room temperature, typically in the range 18-25°C, under anhydrous conditions using HPLC grade solvents. Evaporation was performed by rotary evaporation in vacuum. Column chromatography was performed on silica gel by the flash method. Nuclear magnetic resonance (NMR) shift values were recorded on a Bruker Avance DPX200 or 300, or AVII400 instrument and multiple peaks were described as follows: s, singlet; d, doublet; dd, doublet. Doublet; t, triplet; q, quartet; p, quintet; m, multiplet; br, wide. Mass spectra were recorded using a Gl956A mass spectrometer (electrospray, 3000V), switching between positive and negative ionization modes. The reported yields are illustrative and do not necessarily represent the maximum yields that can be achieved.

[Example 1]
Preparation of tert-butyl 2-((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaen-1-yloxy)butanoate:

Tetrabutylammonium chloride (0.55 g, 1.98 mmol) was added to (5Z,8Z,11Z,14Z,17Z)-equosa-5,8,11,14,17-pentaen-1-ol (3.50 in toluene (35 mL). g, 12.1 mmol) at room temperature under nitrogen. Aqueous sodium hydroxide solution (50% (w/w), 11.7 mL) was added at room temperature with vigorous stirring, followed by t-butyl 2-bromobutyrate (5.41 g, 24.3 mmol). The resulting mixture was heated to 50° C. and additional t-butyl 2-bromobutyrate was added after 1.5 h (2.70 g, 12.1 mmol), 3.5 h (2.70 g, 12.1 mmol) and 4.5 h (2.70 g). g, 12.1 mmol) was added and stirred for a total of 12 hours. After cooling to room temperature, ice water (25 mL) was added and the two phases obtained were separated. The organic phase was washed with NaOH (5%) and a mixture of brine, dried (MgSO _4), filtered, and concentrated. The residue was purified by flash chromatography on silica gel using an increasingly polar mixture of heptane and ethyl acetate (100:0 to 95:5) as eluent. Concentration of the appropriate fractions gave 1.87 g (36% yield) of the title compound as an oil. ¹ H NMR (300 MHz, CDCI3): δ 0.85-1.10 (m, 6H), 1.35-1.54 (m, 11H), 1.53-1.87 (m, 4H), 1.96-2.26 (m, 4H), 2.70-3.02 (m, 8H), 3.31 (dt, 1H), 3.51-3.67 (m, 2H), 5.10-5.58 (m, 10H).

[Example 2]
Preparation of 2-((5Z,8Z,11Z,14Z,17Z)-Icosa-5,8,11,14,17-pentaenyloxy)butanoic acid (Compound A):

tert-Butyl 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaen-1-yloxy)butanoate (19.6 g, 45.5 mmol) in dichloromethane (200 mL) Dissolved and placed under nitrogen. Trifluoroacetic acid (50 mL) was added and the reaction mixture was stirred at room temperature for 1 hour. Water was added and the aqueous phase was extracted twice with dichloromethane. The combined organic extracts were washed with brine, dried (Na ₂ SO ₄ ), filtered and concentrated. The residue was flash chromatographed on silica gel using a mixture of increasingly polar heptane, ethyl acetate and formic acid (90:10:1 to 80:20:1) as eluent. Concentration of the appropriate fractions gave 12.1 g (71% yield) of the title compound as an oil. ¹ H-NMR (300 MHz, CDCl ₃ ): δ 0.90-1.00 (m, 6H), 1.50 (m, 2H), 1.70 (m, 2H), 1.80 (m, 2H), 2.10 (m, 4H), 2.80-2.90 (m, 8H), 3.50 (m, 1H), 3.60 (m, 1H), 3.75 (t, 1H), 5.30-5.50 (m, 10H); MS (Electrospray): 373.2 [MH] ^- .

[Example 3]
(4S,5R)-3-((S)-2-((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaenyloxy)butanoyl)-4-methyl -5-Phenyloxazolidin-2-one and (4S,5R)-3-((R)-2-((5Z,8Z,11Z,14Z,17Z)-equosa-5,8,11,14,17- Preparation of pentaenyloxy)butanoyl)-4-methyl-5-phenyloxazolidin-2-one:

  DMAP (1.10 g, 8.90 mmol) and DCC (1.90 g, 9.30 mmol) in 2-((5Z,8Z,11Z,14Z,17Z)-Icosa-5,8,11,14 in dry dichloromethane (100 mL). A mixture of 17,17-pentaenyloxy)butanoic acid (3.20 g, 8.50 mmol) was added and kept at 0° C. under nitrogen. The resulting mixture was stirred at 0°C for 20 minutes. (4S,5R)-4-Methyl-5-phenyloxazolidin-2-one (1.50 g, 8.50 mmol) was added and the resulting cloudy mixture was stirred at ambient temperature for 5 days. The mixture was filtered and concentrated under reduced pressure to give a crude product containing the desired product as a mixture of two diastereomers. The residue was purified by flash chromatography on silica gel using 15% ethyl acetate in heptane as eluent. The two diastereomers were separated and the appropriate fractions were concentrated. (4S,5R)-3-((S)-2-((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaenyloxy)butanoyl)-4-methyl The -5-phenyloxazolidin-2-one was eluted first to give 1.1 g (40% yield) as an oil. (4S,5R)-3-((R)-2-((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaenyloxy)butanoyl)-4-methyl 0.95 g (34% yield) of -5-phenyloxazolidin-2-one was obtained as an oil.

(4S,5R)-3-((S)-2-((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaenyloxy)butanoyl)-4-methyl -5-Phenyloxazolidin-2-one (E1): ¹ H-NMR (300 MHz, CDCl ₃ ): δ 0.90 (d, 3H), 1.00 (t, 3H), 1.07 (t, 3H), 1.45-1.57 (m, 2H), 1.62-1.76 (m, 3H), 1.85-1.95 (m, 1H), 2.05-2.15 (m, 4H), 2.87 (m, 8H), 3.39 (m, 1H), 3.57 (m , 1H), 4.85-4.92 (m, 2H), 5.30-5.45 (m, 10H), 5.75 (d, 1H), 7.32 (m, 2H), 7.43 (m, 3H).

(4S,5R)-3-((R)-2-((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaenyloxy)butanoyl)-4-methyl -5-Phenyloxazolidin-2-one (E2): ¹ H-NMR (300 MHz, CDCl ₃ ): δ 0.98 (d, 3H), 0.99 (t, 3H), 1.08 (t, 3H), 1.40-1.52 (m, 2H), 1.55-1.75 (m, 3H), 1.80-1.90 (m, 1H), 2.05-2.15 (m, 4H), 2.84 (m, 8H), 3.39 (m, 1H), 3.56 (m , 1H), 4.79 (quintet, 1H), 4.97 (dd, 1H), 5.30-5.45 (m, 10H), 5.71 (d, 1H), 7.33 (m, 2H), 7.43 (m, 3H).

[Example 4]
Preparation of (S)-2-((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaenyloxy)butanoic acid:

Hydrogen peroxide (35% in water, 0.75 mL, 8.54 mmol) and lithium hydroxide monohydrate (0.18 g, 4.27 mmol) in tetrahydrofuran (12 mL) and water (4 mL) (4S,5R)-3- ((S)-2-((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaenyloxy)butanoyl)-4-methyl-5-phenyloxazolidine-2- It was added to a solution of on (1.10 g, 2.13 mmol) and kept at 0°C under nitrogen. The reaction mixture was stirred at 0° C. for 30 minutes. 10% Na ₂ SO ₃ (aq) (30 mL) was added, the pH was adjusted to about 2 with 2M HCl and the mixture was extracted twice with heptane (30 mL). The combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated. The residue was subjected to flash chromatography on silica gel using an increasingly polar mixture of heptane and ethyl acetate (98:8 to 1:1) as eluent. Concentration of appropriate fractions gave 0.48 g (60% yield) of the title compound as an oil. ¹ H-NMR (300 MHz, CDCl ₃ ): δ 0.90-1.00 (m, 6H), 1.48 (m, 2H), 1.65 (m, 2H), 1.85 (m, 2H), 2.10(m, 4H), 2.80-2.90 (m, 8H), 3.55 (m, 1H), 3.60 (m, 1H), 3.88 (t, 1H), 5.35-5.45 (m, 10H); MS (Electrospray): 373.3 [MH] ^- ; [α] _D -37° (c=0.104, ethanol).

[Example 5]
Preparation of (R)-2-((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaenyloxy)butanoic acid:

Hydrogen peroxide (35% in water, 0.65 mL, 7.37 mmol) and lithium hydroxide monohydrate (0.15 g, 3.69 mmol) in tetrahydrofuran (12 mL) and water (4 mL) (4S,5R)-3- ((R)-2-((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaenyloxy)butanoyl)-4-methyl-5-phenyloxazolidine-2- It was added to a solution of on (0.95 g, 1.84 mmol) and kept at 0° C. under nitrogen. The reaction mixture was stirred at 0° C. for 30 minutes. 10% Na ₂ SO ₃ (aq) (30 mL) was added, the pH was adjusted to 2 with 2M HCl and the mixture was extracted twice with heptane (30 mL). The combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated. The residue was subjected to flash chromatography on silica gel using an increasingly polar mixture of heptane and ethyl acetate (98:8 to 50:50) as eluent. Concentration of the appropriate fractions gave 0.19 g (29% yield) of the title compound as an oil. ¹ H-NMR (300 MHz, CDCl ₃ ): δ 0.90-1.00 (m, 6H), 1.48 (m, 2H), 1.65 (m, 2H), 1.85 (m, 2H), 2.10 (m, 4H), 2.80-2.90 (m, 8H), 3.55 (m, 1H), 3.60 (m, 1H), 3.88 (t, 1H), 5.35-5.45 (m, 10H); MS (Electrospray): 373.3 [MH] ^- ; [α] _D -31° (c=0.088, ethanol).

[Example 6]
Preparation of tert-butyl 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)propanoate:

(5Z,8Z,11Z,14Z,17Z)-Icosa-5,8,11,14,17-pentaen-1-ol (1.00 g, 3.47 mmol), tetrabutylammonium chloride (0.24 g, 0.87 mmol) and t A mixture of -butyl 2-bromopropionate (3.62g, 17.3mmol) was dissolved in toluene (36mL) and placed under nitrogen. An aqueous solution of sodium hydroxide (50%, 8 mL) was added slowly with vigorous stirring and the resulting mixture was stirred at ambient temperature for 20 hours. Water was added and the mixture was extracted 3 times with ether. The combined organic extracts were washed with brine, dried (Na ₂ SO ₄ ), filtered and concentrated. The residue was purified by flash chromatography on silica gel using 2% ethyl acetate in heptane as the eluent. Concentration of the appropriate fractions gave 1.40 g (90% yield) of the title compound as an oil. ¹ H-NMR (300 MHz, CDCl ₃ ): δ 0.95 (t, 3H), 1.41 (d, 3H), 1.48 (s, 9H), 1.48-1.66 (m, 4H), 2.05 (m, 4H), 2.83 (m, 8H), 3.35 (m, 1H), 3.55 (m, 1H), 3.79 (q, 1H), 5.32-5.44 (m, 10H).

[Example 7]
Preparation of 2-((5Z,8Z,11Z,14Z,17Z)-equosa-5,8,11,14,17-pentaenyloxy)propanoic acid:

Trifluoroacetic acid (2 mL) was added to 2-((5Z,8Z,11Z,14Z,17Z)-equosa-5,8,11,14,17-pentaenyloxy)propanoate (1.40 g, in dichloromethane (10 mL). 3.36 mmol) and kept under nitrogen and the reaction mixture was stirred at room temperature for 3 hours. Diethyl ether (50 mL) was added and the organic phase was washed with water (30 mL), dried (Na ₂ SO ₄ ) and concentrated. The residue was flash chromatographed on silica gel using a mixture of increasingly polar heptane, ethyl acetate and formic acid (95:5:0.25 to 80:20:1) as eluent. Concentration of the appropriate fractions gave 0.67 g of slightly impure product. This material was dissolved in heptane (15 mL), washed 3 times with water (5 mL), dried (Na ₂ SO ₄ ), filtered and concentrated to give 0.50 g (41% yield) of the title as an oil. The compound was obtained. ¹ H-NMR (300 MHz, CDCl ₃ ): δ 0.99 (t, 3H), 1.40-1.48 (m, 5H), 1.67 (m, 2H), 2.09 (m, 4H), 2.80-2.60 (m, 8H ), 3.53 (m, 2H), 4.01 (q, 1H), 5.31-5.47 (m, 10H); MS (Electrospray): 359.2 [MH] ^- .

[Example 8]
Preparation of tert-butyl 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)-2-methylpropanoate:

(5Z,8Z,11Z,14Z,17Z)-Icosa-5,8,11,14,17-pentaen-1-ol (0.83 g, 3.14 mmol), tetrabutylammonium chloride (0.24 g, 0.85 mmol) and t A mixture of -butyl 2-bromoisobutyrate (3.50 g, 15.7 mmol) was dissolved in toluene (15 mL) and placed under nitrogen. Aqueous sodium hydroxide solution (50%, 5 mL) was added at room temperature with vigorous stirring. The resulting mixture was heated to 60° C. and stirred for 6 hours. The mixture was cooled, water was added and extracted 3 times with ether. The combined organic extracts were washed with brine, dried (Na ₂ SO ₄ ), filtered and concentrated. The residue was purified by flash chromatography on silica gel using a 5-10% gradient of ethyl acetate in heptane as the eluent. Concentration of appropriate fractions gave 0.60 g (44% yield) of the title compound as an oil. MS (electrospray): 453.3 [M+Na] ⁺ .

[Example 9]
Preparation of 2-((5Z,8Z,11Z,14Z,17Z)-equosa-5,8,11,14,17-pentaenyloxy)-2-methylpropanoic acid:

Trifluoroacetic acid (5 mL) was added to tert-butyl 2-((5Z,8Z,11Z,14Z,17Z)-equosa-5,8,11,14,17-pentaenyloxy)-2 in dichloromethane (20 mL). -Methyl propanoate (600 mg, 1.39 mmol) was added under nitrogen and the reaction mixture was stirred at room temperature for 2 hours. Water was added and the aqueous phase was extracted twice with dichloromethane. The combined organic extracts were washed with brine, dried (Na ₂ SO ₄ ), filtered and concentrated. The residue was purified by flash chromatography on silica gel using a mixture of heptane, ethyl acetate and formic acid (80:20:1) as eluent. The appropriate fractions were concentrated and the residue (135 mg) was further flash chromatographed on silica gel using a 5-10% gradient of a mixture of ethyl acetate and formic acid in heptane (95:5) as eluent. Purified. Concentration of the appropriate fractions gave 80 mg of slightly impure product. This material was dissolved in heptane (5 mL), washed twice with water (5 mL), dried (Na ₂ SO ₄ ), filtered and concentrated to 40 mg (8% yield) of the title compound as an oil. Got ¹ H-NMR (300 MHz, CDCl ₃ ): δ 0.99 (t, 3H), 1.47 (s, 6H), 1.64 (m, 2H), 2.07 (m, 4H), 2.81-2.88 (m, 8H), 3.46 (t, 2H), 5.29-5.44 (m, 10H); MS (Electrospray): 373.3 [MH] ^- .

[Example 10]
Preparation of tert-butyl 2-ethyl-2-((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaen-1-yloxy)butanoate:

tert-Butyl 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaen-1-yloxy)butanoate (480 mg, 1.11 mmol) in dry tetrahydrofuran (10 mL) A solution of lithium diisopropylamine (LDA) (2.0M, 750 μL, 1.50 mmol) in was added dropwise over 30 minutes and kept at -70°C under nitrogen. The reaction mixture was stirred for 30 minutes. Ethyl iodide (312 mg, 2.00 mmol) was added in one portion and the resulting mixture was warmed to ambient temperature for 1 hour. The reaction mixture was stirred at ambient temperature for 17 hours. The mixture was poured into saturated NH ₄ Cl (aq) (50mL), (2 × 50mL) heptane extracted. The combined organic phases brine (50 mL), washed successively with 0.25M of HCl (50 mL) and brine (50 mL), dried (MgSO _4), filtered, and concentrated. The residue was purified by flash chromatography on silica gel using an increasingly polar mixture of heptane and ethyl acetate (100:0 to 95:5) as eluent. Concentration of the appropriate fractions gave 343 mg (67% yield) of the title compound as an oil. ¹ H NMR (300 MHz, CDCI ₃ ): δ 0.84 (t, 6H), 0.99 (td, 3H), 1.35-1.55 (m, 11 H), 1.54-1.69 (m, 2H), 1.68-1.87 (m , 4H), 1.99-2.24 (m, 4H), 2.74-2.99 (m, 8H), 3.31 (t, 2H), 5.23-5.52 (m, 10H); MS (Electrospray): 401.3 [M-1] ^- .

[Example 11]
Preparation of 2-ethyl-2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaen-1-yloxy)butanoic acid:

Formic acid (5 ml) and tert-butyl 2-ethyl-2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaen-1-yloxy)butanoate (250 mg, 0.55 mmol) was stirred vigorously under nitrogen at room temperature for 4.5 hours. Formic acid was removed in vacuo. The residue was purified by flash chromatography on silica gel using a mixture of increasingly polar heptane and ethyl acetate (100:0 to 80:20) as eluent. Concentration of the appropriate fractions gave 163 mg (74% yield) of the title compound as an oil. ¹ H NMR (300 MHz, CDCl ₃ ): δ 0.86 (t, 6H), 0.99 (t, 3H), 1.36-1.57 (m, 2H), 1.68 (dd, 2H), 1.73-1.98 (m, 4H) , 2.11 (tt, 4H), 2.70-3.01 (m, 8H), 3.39 (t, 2H), 5.20-5.56 (m, 10H). MS (Electrospray): 481.4 [M+Na] ⁺ .

[Example 12]
Preparation of ethyl 2-(((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoate:

Dicyclohexylmethanediimine (DCC) (304 mg, 1.47 mmol) and N,N-dimethylpyridin-4-amine (DMAP) (10 mg, 0.067 mmol) were added to 2-(((5Z ,8Z,11Z,14Z,17Z)-Icosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoic acid (501.3 mg, 1.335 mmol) in a stirred solution at 0° C., nitrogen Added under atmosphere. After the mixture was stirred for 5 minutes, ethanol (EtOH) (0.16 mL, 2.67 mmol) was added. The resulting mixture was stirred at room temperature for 20 hours. The reaction mixture was purified by flash chromatography on silica gel using an increasing mixture of heptane and ethyl acetate (100:0 to 99:1) as eluent. Concentration of the appropriate fractions gave 473 mg (88% yield) of the title compound as an oil. ¹ H NMR (300 MHz, chloroform-d) δ 0.95 (2 xt, 6H), 1.37-1.48 (m, 2H), 1.54-1.79 (m, 4H), 2.01-2.10 (m, 4H), 2.77-2.84 (m, 8H), 3.27-3.34 (m, 1H), 3.53-3.60 (m, 1H), 3.69-3.73 (dd, 1H), 4.13-4.24 (m, 2H), 5.25-5.33 (m, 10H) , MS (electrospray); 425.3 [M+Na] ⁺ ; HRMS (electrospray): found 425.3021 [M+Na] ⁺ , calculated [C ₂₆ H ₄₂ O ₃ +Na] ⁺ 425.3303.

[Example 13]
Preparation of isopropyl 2-(((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoate

DCC (310 mg, 1.47 mmol) and DMAP (9 mg, 0.067 mmol) were added to 2-(((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17 in DCM (4 mL). To the stirred solution of -pentaen-1-yl)oxy)butanoic acid (501 mg, 1.335 mmol) was added at 0°C under nitrogen atmosphere. The mixture was stirred for 5 minutes and then isopropanol (iPrOH) (0.16 mL, 2.67 mmol) was added. The resulting mixture was stirred at room temperature for 20 hours. The mixture was filtered and concentrated in vacuo. The residue was added to heptane (50 mL), filtered and concentrated in vacuo. The residue was purified by flash chromatography on silica gel using 1% ethyl acetate in heptane as eluent. Concentration of the appropriate fractions gave 496 mg (89% yield) of the title compound as an oil. ¹ H NMR (300 MHz, CDCl ₃ ): δ 0.97 (2 xt, 6H), 1.25 (m, 6H), 1.42-1.50 (m, 2H), 1.61-1.70 (m, 2H), 1.70-1.77 (m , 2H), 2.05-2.12 (m, 4H), 2.79-2.86 (m, 8H), 3.29-3.34 (m, 1H), 3.54-3.59 (m, 1H), 3.67-3.71 (m, 1H), 5.06 -5.10 (m, 1H), 5.31-5.42 (m, 10 H); MS (Electrospray): 439.3 [M+Na] ⁺ .

[Example 14]
Preparation of methyl 2-(((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoate:

Sulfuric acid (0.049 ml, 0.918 mmol) was added to 2-(((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaen-1-yl) in methanol (20 ml). To a solution of (oxy)butanoic acid (385 mg, 1.028 mmol) was added at room temperature under N ₂ atmosphere and the resulting mixture was stirred at room temperature overnight. MS (Electrospray): 389.3 [M+1] ⁺ .

[Example 15]
Preparation of butyl 2-(((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoate:

Sulfuric acid (0.049 ml, 0.918 mmol) was added to 2-(((5Z,8Z,11Z,14Z,17Z)-equosa-5,8,11,14,17 in butan-1-ol (400 mL, 4.37 mol). To the solution of -pentaen-1-yl)oxy)butanoic acid (33 g, 88 mmol) was added at room temperature under N ₂ atmosphere and the reaction mixture was stirred for 120 h. Heptane (400 mL) and ethyl acetate (400 mL) were added and the solution was washed with saturated aqueous NaHCO ₃ (3 x 300 mL) and water (2 x 300 mL). The combined aqueous phases were extracted with heptane/ether (1:1) (2 x 300 mL). The combined organic phases were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. The residue was purified by flash chromatography using an increasing mixture of heptane and ethyl acetate (99:1 to 96:4) as eluent. Concentration of the appropriate fractions gave 26.3 g (67% yield) of the title compound as an oil. ¹ H NMR (400 MHz, CDCl3) δ 0.93-1.02 (m, 9H), 1.36-1.51 (m, 4H), 1.60-1.70 (m, 4H), 1.72-1.84 (m, 2H), 2.05-2.16 ( m,4H), 2.78-2.92 (m, 8H), 3.28-3.39 (m, 1H), 3.54-3.65 (m, 1H), 3.70-3.82 (m, 1H), 4.08-4.24 (m, 2H), 5.27-5.48 (m, 10H), MS (Electrospray): 453.2 [M+Na] ⁺ .

[Example 16]
Preparation of 2,3-dihydroxypropyl 2-(((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoate:

  Step a) (2,2-dimethyl-1,3-dioxolan-4-yl)methyl 2-(((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaene Preparation of 1-yl)oxy)butanoate

2-(((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoic acid (25 g, 66.7 mmol) and DMAP (8.15 g, 66.7 mmol) was added to a solution of 2,2-dimethyl-1,3-dioxolane-4-methanol (7.54 mL, 60.7 mmol) in chloroform (150 mL) under nitrogen atmosphere. Then a solution of DCC (13.77 g, 66.7 mmol) in chloroform (65 mL) was added dropwise at ambient temperature. The mixture was stirred overnight and concentrated in vacuo. The residue was purified by flash chromatography on silica gel using a mixture of 10% ethyl acetate in heptane of increasing polarity as eluent. Concentration of the appropriate fractions gave 19.6 g (66% yield) of the title product as an oil. ¹ H NMR (300 MHz, CDCl3) δ 0.99 (t, 6H), 1.37-1.40 (m, 3H), 1.41-1.53 (m, 5H), 1.59-1.71 (m, 2H), 1.72-1.85 (m, 2H), 2.05-2.14 (m, 4H), 2.74-2.95 (m, 8H), 3.31-3.38 (m, 1H), 3.57-3.65 (m, 1H), 3.72-3.86 (m, 2H), 4.07- 4.12 (m, 1H), 4.15-4.27 (m, 2H), 4.29-4.40 (m, 1H), 5.23-5.50 (m, 10H). MS (Electrospray): 511.3 [M+Na] ⁺ .

  Step b) Preparation of 2,3-dihydroxypropyl 2-(((5Z,8Z,11Z,14Z,17Z)-equosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoate

(2,2-Dimethyl-1,3-dioxolan-4-yl)methyl 2-(((5Z,8Z,11Z,14Z,17Z)-equosa-5,8,11,14, in dioxane (280 mL) To a solution of 17-pentaen-1-yl)oxy)butanoate (27.5 g, 56.3 mmol) was added aqueous HCl (37% (w/w), 28 mL, 341 mmol) at room temperature under nitrogen and the mixture was added to 60%. Stir for minutes. The mixture was then carefully poured into saturated aqueous NaHCO ₃ (500 mL) and extracted with EtOAc (2 x 300 mL). The organic phase was washed with 1M HCl (200 mL), brine (200 mL), dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. The residue was purified by flash chromatography on silica gel using heptane and ethyl acetate (50:50) as eluent. Concentration of the appropriate fractions gave 19 g of the title product as an oil. The product contained approximately 10% of the isomer 1,3-dihydroxypropan-2-yl 2-(((5Z,8Z,11Z,14Z,17Z)-equosa-5,8,11,14,17-pentaene. 1-yl)oxy)butanoate was mixed. This material was mixed with another batch of 1.35 grams and then further purified by preparative HPLC. A 17:83 homogeneous mixture of water/acetonitrile (9:1) to acetonitrile (100%) was used as the eluent. Concentration of the appropriate fractions gave 11.3 g (38% yield) of the title product as an oil. ¹ H NMR (300 MHz, CDCl3) δ 0.97-1.03 (m, 6H), 1.41-1.51 (m, 2H), 1.59-1.69 (m, 2H), 1.72-1.87 (m, 2H), 2.05-2.14 ( m, 5H), 2.56 (s, 1H), 2.73-2.94 (m, 8H), 3.33-3.40 (m, 1H), 3.55-3.68 (m, 2H), 3.69-3.77 (m, 1H), 3.79- 3.85 (m, 1H), 3.93-4.03 (m, 1H), 4.15-4.37 (m, 2H), 5.25-5.51 (m, 10H). MS (Electrospray): 471.3 [M+Na] ⁺ .

[Example 17]
Preparation of 1,3-dihydroxypropan-2-yl 2-(((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoate

  Step a) Preparation of oxiran-2-ylmethyl 2-(((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoate

2-(((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoic acid (800 mg, 2.14 mmol) in dry DCM (7 mL). ), glycidol (0.17 mL, 2.56 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC*HCl) (491 mg, 2.56 mmol) and 4-dimethylaminopyridine (DMAP) (313 mg, A mixture of 2.56 mmol) was stirred at room temperature under N ₂ atmosphere. The reaction mixture was concentrated in vacuo. The residue was purified by flash chromatography on silica gel using an increasingly polar mixture of heptane and ethyl acetate (99:1 to 95:5) as eluent. Concentration of the appropriate fractions gave 527 mg (57% yield) of the title product as an oil. ¹ H NMR (400 MHz, CDCl3) δ 0.94-0.98 (m, 6H), 1.40-1.44 (m, 2H), 1.57-1.64 (m, 2H), 1.70-1.82 (m, 2H), 2.02-2.12 ( m, 4H), 2.63 (bs, 1H), 2.78-2.84 (m, 9H), 3.20 (bs, 1H), 3.30-3.35 (m, 1H), 3.55-3.61 (m, 1H), 3.77-3.80 ( m, 1H), 3.94-4.01 (m, 1H), 4.42-4.48 (m, 1H), 5.36-5.26 (m, 10H). MS (electrospray): 453.3 [M+Na] ⁺ .

  Step b) 2-((2-(((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoyl)oxy)propane-1 Preparation of 1,3-diylbis(2,2,2-trifluoroacetate)

Trifluoroacetic anhydride (TFAA) (0.55 mL, 3.96 mmol) in dry DCM (3 mL) was added to oxiran-2-ylmethyl 2-(((5Z,8Z,11Z,14Z,17Z )-Icosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoate (286 mg, 0.66 mmol) in a pre-cooled solution at -20°C under N ₂ atmosphere in several portions. Was added. The cooling bath was removed and the mixture was stirred for 19 hours at ambient temperature before the reaction mixture was concentrated under reduced pressure. The residue was dissolved in toluene (6 mL), passed through a pad of silica (6.5 g) and eluted with toluene (150 mL). Concentration in vacuo gave 357 mg (84% yield) of the title compound as an oil. ¹ H NMR (400 MHz, CDCl3) δ 0.95 (2xt, 6H), 1.38-1.45 (m, 2H), 1.57-1.63 (m, 2H), 1.66-1.78 (m, 2H), 2.09-2.02 (m, 4H), 2.78-2.84 (m, 8H), 3.27-3.33 (m, 1H), 3.51-3.56 (m, 1H), 3.77 (dd, 1H), 4.30-4.53 (m, 2H), 4.60-4.69 ( m, 2H), 5.17-5.43 (m, 10H), 5.43-5.55 (m, 1H). MS (electrospray): 661.1 [M+Na] ⁺ .

  Step c) 1,3-dihydroxypropan-2-yl 2-(((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoate Preparation of

A solution of pyridine (0.4 mL, 4.95 mmol) and methanol (0.3 mL, 7.41 mmol) in pentane/DCM (2:1) (4.5 mL) was treated with 2- in pentane/DCM (2:1) (5 mL). ((2-(((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoyl)oxy)propane-1,3-diylbis( To a solution of (2,2,2-trifluoroacetate) (354 mg, 0.552 mmol) was cooled to -20°C and added dropwise under N ₂ atmosphere. The cooling bath was removed and the mixture was stirred for 3 hours at room temperature then concentrated in vacuo. The residue was purified by flash chromatography on silica gel using an increasingly polar mixture of heptane and ethyl acetate (95:5 to 90:10, 80:20, 50:50) as eluent. Concentration of the appropriate fractions gave 223 mg of the title product as a crude oil. Purification by preparative HPLC gave 58 mg (22% yield) of the title compound as an oil. ¹ H NMR (400 MHz, CDCl3) δ 0.95 (t, 3H), 0.96 (t, 3H), 1.38-1.45 (m, 2H), 1.54-1.64 (m, 2H), 1.67-1.84 (m, 2H) , 2.01-2.09 (m, 4H), 2.45 (bs, 2H), 2.83-2.77 (m, 8H), 3.36-3.30 (m, 1H), 3.60-3.55 (m, 1H), 3.84-3.78 (m, 5H), 4.98-4.93 (m, 1H), 5.65-5.09 (m, 10H). MS (electrospray): 471.1 [M+Na] ⁺ .

[Example 18]
3-hydroxypropane-1,2-diylbis(2-(((5Z,8Z,11Z,14Z,17Z)-equosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoate) Preparation

  Step a) Preparation of tert-butyl ((2,2-dimethyl-1,3-dioxolan-4-yl)methoxy)dimethylsilane

tert-Butyl-chlorodimethylsilane (14.41 g, 91 mmol), in THF (100 mL) (2,2-dimethyl-1,3-dioxolan-4-yl)methanol (10 g, 76 mmol) and imidazole (7.73 g, 114 mmol) was added to the solution at ambient temperature under nitrogen atmosphere. The mixture was stirred for 1.5 hours, poured into water (200 mL) and extracted with tert-butyl methyl ether (2 x 150 mL). The phases were separated and the organic layer was washed with water (100 mL), brine (100 mL), dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. The residue was purified by flash chromatography on silica gel using 3% ethyl acetate in heptane as the eluent. Concentration of the appropriate fractions gave 18 g (97% yield) of the title compound as an oil. 1H NMR (300 MHz, CDCl3) δ 0.02-0.05 (m, 6H), 0.85-0.89 (m, 9H), 1.31-1.35 (m, 3H), 1.35-1.40 (m, 3H), 3.50-3.60 (m , 1H), 3.63-3.72 (m, 1H), 3.75-3.85 (m, 1H), 3.96-4.05 (m, 1H), 4.07-4.18 (m, 1H). MS (electrospray): 229.2 [M+ Na] ⁺ .

  Step b) Preparation of 3-((tert-butyldimethylsilyl)oxy)propane-1,2-diol

In a solution of tert-butyl ((2,2-dimethyl-1,3-dioxolan-4-yl)methoxy)dimethylsilane in chloroform (60 mL), FeCl ₃ x6H ₂ O absorbed on silica gel (30 g, 11.9 mmol) was added and the mixture was stirred overnight. The mixture was filtered and concentrated in vacuo. The residue was purified by flash chromatography on silica gel using a mixture of increasingly polar heptane and ethyl acetate (50:50 to 25:75) as eluent. Concentration of the appropriate fractions gave 760 mg (9% yield) of the title compound as an oil. ¹ H NMR (300 MHz, CDCl3) δ 0.09-0.12 (m, 6H). 0.91- 0.95 (m, 9H), 2.11-2.17 (m, 1H), 2.60 (d, 1H), 3.57- 3.85 (m, 5H). MS (electrospray): 229.2 [M+Na] ⁺ .

  Step c) 3-((tert-butyldimethylsilyl)oxy)propane-1,2-diylbis(2-(((5Z,8Z,11Z,14Z,17Z)-equosa-5,8,11,14,17 -Pentaen-1-yl)oxy)butanoate)

To a solution of 3-((tert-butyldimethylsilyl)oxy)propane-1,2-diol (0.91 g, 4.41 mmol) in DMF (20 ml) under N ₂ atmosphere at ambient temperature, 2-((( 5Z,8Z,11Z,14Z,17Z)-Icosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoic acid (3.47 g, 9.3 mmol), DMAP (1.13 g, 9.3 mmol), 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (DCI) (1.776 g, 9.26 mmol) and dry DCM (60 ml) were added. After stirring the mixture overnight, the reaction mixture was diluted with diethyl ether (200 mL). The mixture was washed with 1M HCl (100 mL) and brine (100 mL), dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. The residue was purified by flash chromatography on silica gel using 3% ethyl acetate in heptane as the eluent. Concentration of the appropriate fractions gave 2.26 g (56% yield) of the title compound as an oil. ¹ H NMR (300 MHz, CDCl3) δ 0.08 (s, 6H), 0.90 (d, 9H), 0.95-1.03 (m, 12H), 1.40-1.52 (m, 4H), 1.58-1.69 (m, 4H) , 1.70-1.83 (m, 4H), 2.04-2.15 (m, 8H), 2.77-2.92 (m, 16H), 3.27-3.37 (m, 2H), 3.57-3.67 (m, 2H), 3.72-3.80 ( m, 4H), 4.14-4.32 (m, 1H), 4.41-4.56 (m, 1H), 5.09-5.22 (m, 1H), 5.23-5.49 (m, 20H). MS (Electrospray): 941.6 [M +Na] ⁺ .

  Step d) 3-hydroxypropane-1,2-diylbis(2-(((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaen-1-yl)oxy) Butanoate) preparation

3-((tert-butyldimethylsilyl)oxy)propane-1,2-diylbis(2-(((5Z,8Z,11Z,14Z,17Z)-equosa-5,8,11, in dioxane (100 mL) To a solution of 14,17-pentaen-1-yl)oxy)butanoate) (2.26 g, 2.46 mmol) was added aqueous HCl (37% (w/w, 2 mL) and the mixture was stirred for 3 hours under a nitrogen atmosphere. After stirring at ambient temperature, concentrated in vacuo The residue was purified by flash chromatography on silica gel using 15% ethyl acetate in heptane as eluent. Concentration of the appropriate fractions gave an oil. as the title compound 0.83 g (42% ^{yield). 1 H NMR (300 MHz} , CDCl3) δ 0.96-1.03 (m, 12H), 1.40-1.53 (m, 4H), 1.58-1.68 (m, 4H), 1.70-1.85 (m, 4H), 1.87-2.01 (m, 1H), 2.05-2.15 (m, 8H), 2.75-2.95 (m, 16H), 3.28-3.41 (m, 2H), 3.56- 3.65 (m, 2H), 3.73-3.85 (m, 4H), 4.24-4.37 (m, 1H), 4.42-4.53 (m, 1H), 5.14-5.23 (m, 1H), 5.26-5.51 (m, 20H ). MS (Electrospray): 827.5 [M+Na] ⁺ .

[Example 19]
2-Hydroxypropane-1,3-diylbis(2-(((5Z,8Z,11Z,14Z,17Z)-equosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoate) Preparation:

  Step a) 2-oxopropane-1,3-diylbis(2-(((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaen-1-yl)oxy) Butanoate)

2-(((5Z,8Z,11Z,14Z,17Z)-Icosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoic acid (5.0 g, 13.4 mmol) and DMAP (1.63 g) , 13.4 mmol) was added to a solution of 1,3-dihydroxyacetone dimer (1.145 g, 6.36 mmol) in chloroform (25 mL) under nitrogen atmosphere. Then a solution of DCC (2.75 g, 13.35 mmol) in chloroform (10 mL) was added dropwise at ambient temperature. The mixture was stirred overnight at room temperature then concentrated in vacuo. The residue was purified by flash chromatography on silica gel using a mixture of increasingly polar heptane and ethyl acetate (90:10 to 88:12) as eluent. Concentration of the appropriate fractions gave 2.4 g (47% yield) of the title compound as an oil. ¹ H NMR (300 MHz, CDCl3) δ 0.97-1.06 (m, 12H). 1.38-1.53 (m, 4H), 1.57-1.73 (m, 4H), 1.73-1.96 (m, 4H), 2.03-2.17 ( m, 8H), 2.76-2.92 (m, 16H), 3.35-3.42 (m, 2H), 3.63-3.70 (m, 2H), 3.89 (dd, 2H), 4.75-4.93 (m, 4H), 5.27- 5.49 (m, 20H). MS (Electrospray): 827.5 [M+Na] ⁺ .

  Step b) 2-hydroxypropane-1,3-diylbis(2-(((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaen-1-yl)oxy) Butanoate)

NaBH ₄ (0.336 g, 8.87 mmol) was added to 2-oxopropane-1,3-diylbis(2-(((5Z,8Z,11Z,14Z,17Z)-equosa) in THF (55 mL) and water (4 mL). -5,8,11,14,17-Pentaen-1-yl)oxy)butanoate) (3.24 g, 4.03 mmol) was carefully added at 0°C. The mixture was stirred at 0°C for 15 minutes. Then acetic acid (1 mL) was added carefully followed by ethyl acetate (100 mL). The mixture was washed with water (100 mL), saturated aqueous NaHCO ₃ solution (100 mL) and brine before being dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. The residue was combined with another batch of material and then purified by flash chromatography on silica gel using 15% ethyl acetate in heptane as the eluent. Concentration of the appropriate fractions gave 1.62 g (50% yield) of the title compound as an oil. ¹ H NMR (300 MHz, CDCl3) δ 0.97-1.03 (m, 12H), 1.41-1.52 (m, 4H), 1.58-1.69 (m, 6H), 1.71-1.87 (m, 4H), 2.05-2.14 ( m, 8H), 2.38-2.42 (m, 1H), 2.78-2.92 (m, 16H), 3.32-3.39 (m, 2H), 3.57-3.64 (m, 2H), 3.80-3.84 (m, 2H), 4.05-4.34 (m, 5H), 5.26-5.49 (m, 20H). MS (Electrospray): 827.5 [M+Na] ⁺ .

[Example 20]
Preparation of propane-1,2,3-triyltris(2-(((5Z,8Z,11Z,14Z,17Z)-equosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoate)

2-(((5Z,8Z,11Z,14Z,17Z)-Icosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoic acid (4.0 g, 10.7 mmol), 4-dimethylamino Pyridine (1.305 g, 10.7 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.047 g, 10.7 mmol) and dry DCM (30 ml) were added to glycerin (0.173 ml) in DMF (10 ml). , 2.373 mmol) at room temperature under N ₂ atmosphere. After stirring the mixture overnight, the reaction mixture was diluted with diethyl ether (250 mL). The mixture was washed with 1M aq. HCl (100 mL) and brine (100 mL) then dried (Na ₂ SO ₄ ), filtered and evaporated in vacuo. The residue was purified by flash chromatography on silica gel using 5% ethyl acetate in heptane as the eluent. Concentration of the appropriate fractions gave 2.1 g (73% yield) of the title compound as an oil. ¹ H NMR (300 MHz, CDCl3) δ 0.91-1.05 (m, 18H), 1.40-1.52 (m, 6H), 1.57-1.69 (m, 6H), 1.69-1.86 (m, 6H), 2.01-2.17 ( m, 12H), 2.69-2.96 (m, 24H), 3.27-3.38 (m, 3H), 3.53-3.67 (m, 3H), 3.73-3.81 (m, 3H), 4.17-4.27 (m, 2H), 4.37-4.54 (m, 2H), 5.28-5.47 (m, 30H). MS (Electrospray): 1183.8 [M+Na] ⁺ .

[Example 21]
Preparation of calcium 2-(((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoate

2-(((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoic acid (1.87 g, 4.99 mmol, 93%) Mixed with CaCO ₃ (0.25 g, 2.50 mmol). Water (1 ml) was added and the mixture was stirred at RT for 1 h with a mechanical stirrer. CO ₂ is generated. A thick, homogeneous paste was formed. Acetone (7 ml) was added with stirring. Solid material separates. The solid material was filtered, dried over nitrogen, sealed and stored in the refrigerator at 4°C. Yield: 1.86 grams (95%). The solid was not further characterized by analytical or spectroscopic methods, but several experiments were carried out showing that calcium salts were formed:
-Solid material melts below 100°C on a hot plate. No sharp melting point was measured.
The substance does not emit CO ₂ upon addition of an acid, precipitated as "dissolved" and oil.

[Example 22]
Preparation of sodium 2-(((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoate

2-(((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoic acid (1.87 g, 4.99 mmol, 93%) Mixed with NaHCO ₃ (0.420 g, 5.00 mmol). Water (1 ml) was added and the mixture was stirred at RT for 1 h with a mechanical stirrer. CO ₂ was evolved and a thick homogeneous paste was formed. Ethanol (7 ml) was added to the reaction flask with stirring. The sodium salt formed from 2-(((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoic acid is ethanol (7 ml). Is included in the solution when added. A small amount of unreacted NaHCO ₃ was filtered and the solution was evaporated to dryness. The crude slightly viscous oil was evaporated twice with 96% ethanol to remove traces of water.

[Example 23]
2-Hydroxy-N,N,N-trimethylethane-1-aminium 2-(((5Z,8Z,11Z,14Z,17Z)-equosa-5,8,11,14,17-pentaen-1-yl) Preparation of (oxy)butanoate

  Choline hydroxide (327.7 μL) in water was pipetted into a scintillation vial with approximately 2.5 mL MTBE and 7.5 mL n-heptane. In a nitrogen chamber, add 2-(((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoic acid (500 mg, 95.8%). Transferred into vial. In a nitrogen chamber, about 1.0 mL of water was slowly added to the vial with stirring. The vial was then sealed. The reaction mixture was stirred for about 30 minutes. The 2-(((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoic acid choline salt formed was filtered on a Buchner funnel. It was a rigid gel-like substance. The wet material on the filter was washed 3 times with 1 mL MTBET. The washed material appeared to be a rigid gel-like solid.

[Example 24]
Preclinical study Evaluation of apo C-III regulation in a mouse model of dyslipidemia (APOE*3 Leiden transgenic mouse)
The APOE*3Leiden transgenic mouse expresses a mutant form of human apolipoprotein E3 (APOE3), APOE*3Leiden, in addition to human apolipoprotein C1 (APOC1). APOE*3 Leiden transgenic mice show elevated plasma cholesterol and triglyceride levels that are mainly restricted to VLDL/LDL-sized lipoprotein fractions (Van den Maagdenberg AMJM et al., Transgenic mice carrying the apolipoprotein E3-Leiden gene exhibit hyperlipoproteinemia. , J Biol Chem 1993;268:10540-10545). In contrast to normal wild-type mice, APOE*3 Leiden transgenic mice are highly responsive to dietary and antihyperlipidemic agents that affect plasma VLDL and chylomicron levels (Van Vlijmen B et al. , Diet-induced hyperlipoproteinemia and atherosclerosis in apolipoprotein E3-Leiden transgenic mice, J Clin Invest 1994;93:1403-1410; Groot PHE et al., Quantitative assessment of aortic atherosclerosis in apoE3Leiden transgenic mice and its relationship to serum cholesterol exposure,Arterioscler Thromb Vasc Biol 1996;16:926-933). Consequently, this model is suitable for assessing the effects of antihyperlipidemic drugs.

  In this study, female APOE*3 Leiden transgenic mice were used a semi-synthetic western diet (WTD; 15% cocoa butter, 40% sucrose and 0.25% cholesterol; all w/w). After 4 weeks using this diet, plasma cholesterol levels gently reached elevated levels of approximately 12-15 mmol/l. The mice were then subdivided into groups of 10 mice each matched for plasma cholesterol, triglycerides and body weight (t=0). The test substance was 0.3 mmol/kg body weight/day and was mixed with WTD and orally administered to the test. After 4 weeks, all animals were sacrificed and serum and tissues were collected.

  Liver tissue was stored in RNA later (Qiagen) at -80°C. Tissues were homogenized in RLT buffer with dithiothreitol (Qiagen) according to the manufacturer's procedure and RNA was isolated using the RNeasy kit (Qiagen). The quality of the isolated RNA was tested on a Bioanalyser (Agilent) and showed a RIN (RNA integrity value) between 8.1 and 9.5, which represents high quality. cDNA was synthesized by the "RNA to cDNA" kit (Applied Biosystems). Gene expression was measured using a low density array (LDA, specific for mouse RNA (Applied Biosystems)). Each sample was measured in triplicate and results are presented as the average value compared to the control (WTD without addition). Fold change in gene expression was calculated by the comparative Ct method using Rplp0 as the housekeeping gene and the average value of control samples as the standard.

  The results shown in Figure 1 confirm that mice fed Compound A (Example 2) have significantly reduced apo C-III expression than mice fed standard WTD (P< 0.05, Student T-test). The effect of compound A is stronger than the effect of reference compound 12, and the EPA derivative prepared according to Example 20 of WO2010/008299 has the following structure:

  In addition, the ability of both compounds to reduce plasma TG was measured. Both compounds reduced TG levels by 69% compared to controls. This confirms that there is no direct correlation between the observed apo C-III reduction and the TG lowering effect.

[Example 25]
Clinical Studies The apoC-III reducing properties of Compound A were demonstrated in two 12-week studies and one 4-week study in patients with dyslipidemia. All three studies demonstrated a clinically and statistically significant reduction of Apo C-III using Compound A treatment.

[Example 25A]
Population With Severe Hypertriglyceridemia This study investigated patients with fasting plasma triglyceride levels above 500 mg/dL. The primary goal of this study was to evaluate the rate of change of triglyceride (TG) from baseline after 12 weeks of treatment to determine the efficacy of oral administration of Compound A (Example 2) at 600 mg once daily (QD). Was to evaluate. One of the second goals was to assess the impact of Compound A on plasma levels of Apo C-III.

  This Phase II, multicenter, proof-of-concept study included a 6-8 week screening period (including 4 or 6 weeks of diet and lifestyle stabilization/discontinuation and a 2-week TG qualification period). , And a 12-week double-blind, randomized, parallel group, placebo-controlled treatment period.

  After confirmation of qualification of fasting TG levels, suitable subjects entered a 12-week, randomized, double-blind treatment period. At Visit 4 (Week 0), subjects were randomly assigned to one of the following treatment groups in a 1:1 ratio: Compound A 600 mg QD or placebo QD.

  Approximately 43 subjects per treatment group (total of approximately 86 subjects) were randomized in this study. Stratification was by TG level at baseline (≦700 mg/dL or >700 mg/dL), statin use at randomization, and by gender.

  The study population was male and female between ages, including 18-79 years old (female of childbirth had to take suitable measures to avoid pregnancy). Subjects on stabilized lipid-lowering statin treatment and subjects not on non-statin lipid-lowering treatment were eligible to enroll in this study. Subjects must have a mean fasting TG level of ≧500 mg/dL and ≦1500 mg/dL from Visit 2 and Visit 3 values or Visit 3 and Visit 3.1 values prior to randomization Was taken.

  A comprehensive analysis (ITT) population was randomized to receive at least one dose of the investigational product, received a baseline efficacy measurement, and received at least one post-randomization efficacy measurement. It consisted of the subject. The ITT population was the primary analysis population. All efficacy analyzes were performed on the ITT population.

  Summary statistics (n, mean, standard deviation [SD], median, minimum and maximum) for baseline and post-baseline measurements, percent change from baseline, or amount of change were analyzed. All efficacy variables are shown by treatment group and visit.

  The first efficacy analysis was a factor of treatment, gender, and use of statin therapy at randomization, and TG values at baseline as a covariate, using a covariance (ANCOVA) model analysis. Carried out. Least mean squares, standard errors, and two-sided 95% confidence intervals (CI) were obtained for each treatment group and for compound A versus placebo.

  Use ANCOVA model for analysis of secondary efficacy variables, factored by treatment, sex, and use of statin therapy at randomization, and with baseline values for each efficacy variable as covariates did.

  The population recruited for the current study had an average age of 52.5 years and included men (69.0%) and women (31.0%). Approximately 21% of subjects in both treatment groups received statin therapy throughout the study. All other non-statin lipid change therapies were discontinued at screening. Mean compliance rates for studying drug therapy during the study were 96.5% for the placebo group and 99.9% for the 600 mg Compound A group.

  In the ITT population, the least-squares (LS) mean rate of change in apo C-III was -38.0% (-47.5, -28.5) for baseline and -34.7% (-46.5, -22.8) for placebo. Met.

[Example 25B]
Populations with mixed dyslipidemia The present study investigated the use of statins with fasting plasma TG levels between 200 and 499 mg/dL and non-high density lipoprotein cholesterol (non-HDL-C) above 130 mg/dL. Patients who were already on treatment were investigated. The first goal of this study was to evaluate the efficacy of 600 mg QD of Compound A (Example 2) orally by examining the rate of change of triglyceride non-HDL-C from baseline after 12 weeks of treatment. there were. One of the second goals was to assess the impact of Compound A on plasma levels of Apo C-III.

  This Phase II, multicenter, proof-of-concept study included a 6-8 week screening period (4 or 6 weeks diet and lifestyle stabilization/discontinuation period and 2 weeks of TG and non-HDL-C eligibility). (Including the aging period) and a 12-week double-blind, randomized, parallel group, placebo-controlled treatment period.

  After confirmation of qualification of fasting TG and non-HDL-C levels, suitable subjects entered a 12-week, randomized, double-blind treatment period. At Visit 4 (Week 0), subjects were randomly assigned to one of the following treatment groups in a 1:1 ratio: Compound A 600 mg QD or placebo QD.

  The study population was male and female between ages, including 18-79 years old (female of childbirth had to take suitable measures to avoid pregnancy). Subjects on stabilized lipid-lowering statin treatment and subjects not on non-statin lipid-lowering treatment were eligible to enroll in this study. Subjects should have a mean fasting TG level between 200 and 499 mg/dL and a non-over 130 mg/dL level from Visit 2 and Visit 3 or Visit 3 and Visit 3.1 prior to randomization. It was required to have HDL-C values.

  A comprehensive analysis (ITT) population was randomized to receive at least one dose of the investigational product, received a baseline efficacy measurement, and received at least one post-randomization efficacy measurement. It consisted of the subject. The ITT population was the primary analysis population. All efficacy analyzes were performed on the ITT population.

  Summary statistics (n, mean, standard deviation [SD], median, minimum and maximum) for baseline and post-baseline measurements, percent change from baseline, or amount of change were analyzed. All efficacy variables are shown by treatment group and visit.

  The first efficacy analysis was performed using the ANCOVA model, with randomization as a factor and with reference non-HDL-C values as covariates. Least mean squares, standard errors, and two-tailed 95% CIs were obtained for each treatment group and for compound A compared to placebo.

  The first efficacy analysis was based on the 12-week completer population.

  The population recruited for the current study included men (58.4%) and women (46.1%) with an average age of 58.3 years. All subjects were required to be on statin treatment (with or without ezetimibe) during the study. All other non-statin lipid change therapies were discontinued at screening. The average compliance rate for studying drug therapy during the study was 97.2% for the placebo group and 95.3% for the compound A group.

  The baseline mean non-HDL-C level in this study population was 165.9 mg/dL; the baseline median TG level was 262.0 mg/dL.

  In the 12-week completer population, LS mean change rates for apo C-III were -32.5% (-38.4, -26.6) vs. baseline and -20.8% (-28.8, -12.7) vs. placebo. there were.

  Example 25A describes a study in patients with ultra-high triglycerides (TG 500-2000 mg/dl). Example 25B describes a study in statin-stabilized patients with mixed dyslipidemia and persistent hypertriglyceridemia (TG 200-499 mg/dl). The studies included in each section have comparable patient populations and are similar in design.

Example 25C Population with Hypercholesterolemia This study investigated subjects with a fasting LDL-C of at least 2.5 mmol (about 97 mg/dl). The purpose of this study was to determine the pharmacodynamic and hypolipidemic effect of Compound A (Example 2) after 4 weeks of treatment in hypercholesterolemic subjects who discontinued stabilized statin treatment in men. ..

The study population consisted of men aged 18-65 years with a BMI between 18.0 and 35.0 kg/m ² of any ethnic origin.

  The main phase Ib study consisted of a 4-5 week screening period and a 4-week double-blind, randomized, placebo-controlled treatment period.

  All subjects had to have at least 3 months of lipid-lowering statin therapy prior to their first screening visit and at least 4 weeks of stabilized statin prior to their first screening visit ..

  Statin treatment was discontinued at the first screening visit and remained discontinued during the entire screening period. Following discontinuation of statin therapy for at least 21 days, subjects had LDL-C of at least 2.5 mmol/l (about 97 mmol/l) at the second screening visit, prior to randomization and at the first screening visit. There must be at least a 20% increase in LDL-C between the visit and the second screening visit.

  After confirmation of qualification of fasting LDL-C levels, suitable subjects entered a 4-week, double-blind, randomized, placebo-controlled treatment period. Subjects were randomly assigned to one of the following treatment groups at a 3:1 ratio: Compound A 600 mg QD (N=18) or placebo QD (N=6).

  Blood lipids were measured at the end of the screening period and after 4 weeks of treatment. Exploratory pharmacodynamic measurements included LDL-C, VLDL-C, TC, TG, HDL-C, non-HDL-C, and ApoB. The impact of Compound A on Apo C-III was also measured.

  Baseline summary statistics are obtained as means using a coefficient of variation. Mean change from baseline with 95% confidence interval was shown by treatment group for all efficacy variables analyzed.

  The analysis was performed using a covariance (ANCOVA) model analysis with baseline as a covariate for the change from baseline.

The population recruited for the current study included white men (100%) with an average age of 55 years, an average weight of 85 kg, and an average BMI of 27.9 kg/m ² .

  The average percent change in Apo C-III after treatment with Compound A was -42% relative to baseline. This change was statistically significant.

[Example 26]
Relative reduction in Apo C-III achieved by EPA/DHA for Compound A
(a) Effect of EPA/DHA formulations on Compound A on plasma apo C-III and other lipid parameters in subjects with severe HTG
MARINE exam:
In a double-blind, randomized, placebo-controlled study, eicosapentaenoic acid ethyl ester (>96 wt% concentrate) (Vascepa) was administered in 229 subjects with a fasting plasma TG of 500-2000 mg/dl. The effect on Apo C-III was investigated. Vascepa at 4 g/day for 12 weeks reduced median apo C-III levels from 25.6 mg/dl to 19.7 mg/dl, corresponding to a median change of -10.1% from baseline [Journal of Clinical Lipidology 2014;8(3):313-314, Icosapent Ethyl(eicosapentaenoic acid ethyl ester): Effects on Apolipoprotein C-III in patients from the MARINE and ANCHOR studies.] (Table 1).

EVOLVE test:
In a double-blind, randomized, placebo-controlled study, the combination of EPA and DHA as free fatty acids (55 wt% EPA and 20 wt% in 399 patients with a fasting plasma TG of 500-2000 mg/dl). The effect of %DHA) (Epanova) on Apo C-III was investigated. Epanova at 4 g/day for 12 weeks resulted in -15% median change in apo C-III from baseline [Circulation 2012; 126:A19030, Abstract 19030:Apolipoprotein C-III is Significantly Reduced by Prescription Omega-3 Free Fatty Acids (Epanova) in Patients with Severe Hypertriglyceridemia and Changes Correlate with Increases in LDL-C: A Sub-analysis of the EVOLVE trial] (Table 1).

(b) Effect of EPA/DHA formulation against Compound A on plasma apo C-III and other lipid parameters in statin-stabilized subjects with mixed dyslipidemia and persistent hypertriglyceridemia
ANCHOR test:
In a double-blind, randomized, placebo-controlled study, in 702 statin-stabilized patients with mixed dyslipidemia and persistent hypertriglyceridemia with fasting plasma TG of 200-499 mg/dl, The effect of eicosapentaenoic acid ethyl ester (Vascepa) on apo C-III was investigated. Vascepa at 4 g/day for 12 weeks reduced median apo C-III levels from 15.2 mg/dl to 13.7 mg/dl, corresponding to a median change of -9.4% from baseline [Journal of Clinical Lipidology 2015, published, http://dx.doi.org/10.1016/j.jacl.2014.11.009, Effects of icosapent ethyl on lipoprotein particle concentration and size in statin-treated patients with persistent high triglycerides(ANCHOR study)]( Table 2).

ESPRIT test:
In a double-blind, randomized, placebo-controlled study, 647 statin-stabilized patients with mixed dyslipidemia and persistent hypertriglyceridemia with fasting plasma TG of 200-499 mg/dl The effect of a combination of EPA and DHA (Epanova) as free fatty acids on Apo C-III was investigated. Epanova at 4 g/day for 12 weeks resulted in an average apo C-III change from baseline of -13.1% [JACC 2013;61: E1468, A highly bioavailable omega-3 fatty acid reduces. Non-high density lipoprotein cholesterol in high-risk patients treated with a statin and residual hypertriglyceridemia (ESPRIT trial)] (Table 2).

COMBOS test:
In a double-blind, randomized study, EPA and DHA in 256 statin-stabilized patients with mixed dyslipidemia and persistent hypertriglyceridemia with fasting plasma TG of 200-400 mg/dl The effect of the ethyl ester combination (46.5% by weight EPA EE and 37.5% by weight DHA EE) (Lovaza) on Apo C-III was investigated. Lovaza at 4 g/day, week resulted in a -7.8% median change in apoC-III from baseline [Clinical Therapeutics 2007;29(7):1354-1367, Efficiency and tolerability of. adding prescription Omega-3 fatty acids 4 g/d to simvastatin 40 mg/d in hypertriglyceridemic patients: An 8-week, randomized, double-blind, placebo-controlled study] (Table 2).

Summary of Relatively Reduced Plasma Apo C-III with EPA/DHA for Compound A A head-to-head study was not completed, but an equivalent patient population and study design A convincing benchmark is provided to compare the efficacy of Compound A against EPA/DHA in reducing C-III. There are two notable differences between naturally occurring omega-3 fatty acids and Compound A.

  The first factor is the superior efficacy of Compound A which achieved median reductions in apo C-III of 35 and 41% in mixed dyslipidemia and severe HTG patient populations, respectively. This is compared to an apo C-III reduction of only 7.8-15% in the EPA/DHA study.

又はその薬学的に許容される塩若しくはエステルを対象に投与することを含み、
式中、R₁及びR₂は、水素原子又は直鎖、分岐、及び/若しくは環式C₁〜C₆アルキル基から独立して選択され、但しR₁及びR₂が両方とも水素であることはない、方法。
（付記２）
前記化合物が、エナンチオマー、ジアステレオマー、又はこれらの混合物の形態で存在する、付記1に記載の方法。
（付記３）
R₁及びR₂が、水素原子、メチル基、エチル基、n-プロピル基及びイソプロピル基から選択される、付記1に記載の方法。
（付記４）
R₁及びR₂が、水素原子、メチル基、及びエチル基から選択される、付記1に記載の方法。
（付記５）
R₁及びR₂の一方が水素原子であり、R₁及びR₂の他方がC₁〜C₃アルキル基から選択される、付記1に記載の方法。
（付記６）
R₁及びR₂の一方が水素原子であり、R₁及びR₂の他方がメチル基又はエチル基から選択される、付記1に記載の方法。
（付記７）
前記エステルが、グリセリド、及びC₁〜C₆アルキルエステルから選択される、付記1に記載の方法。
（付記８）
前記エステルが、トリグリセリド、1,2-ジグリセリド、1,3-ジグリセリド、1-モノグリセリド、及び2-モノグリセリドから選択される、付記1に記載の方法。
（付記９）
前記エステルが、メチルエステル、エチルエステル、イソプロピルエステル、n-ブチルエステル、及びtert-ブチルエステルから選択される、付記1に記載の方法。
（付記１０）
前記エステルが、メチルエステル及びエチルエステルから選択される、付記1に記載の方法。
（付記１１）
前記化合物が、そのR体で存在する、付記2に記載の方法。
（付記１２）
前記化合物が、そのS体で存在する、付記2に記載の方法。
（付記１３）
前記化合物が、ラセミ体で存在する、付記2に記載の方法。
（付記１４）
R₁が水素であり、R₂がエチルであり、前記式が

である、付記1に記載の方法。
（付記１５）
前記化合物が、式:

で表されるそのS及び/又はR体で存在する、付記14に記載の方法。
（付記１６）
式(I)の化合物の薬学的有効量が、1用量当たり約5mg〜約2gの範囲である、付記1に記載の方法。
（付記１７）
式(I)の化合物の薬学的有効量が、1用量当たり約200mg〜約800mgの範囲である、付記1に記載の方法。
（付記１８）
式(I)の化合物の薬学的有効量が、約600mgである、付記1に記載の方法。
（付記１９）
前記対象が、ヒトである、付記1に記載の方法。
（付記２０）
前記化合物を、1日1回投与する、付記1に記載の方法。
（付記２１）
前記化合物が、経口投与用の医薬組成物として製剤されている、付記1に記載の方法。（付記２２）
前記医薬組成物が、ゼラチンカプセル剤又は錠剤の形態である、付記21に記載の方法。（付記２３）
前記医薬組成物が、少なくとも1種の結合剤、賦形剤、希釈剤、又はこれらの任意の組合せを更に含む、付記22に記載の方法。
（付記２４）
前記医薬組成物が、抗酸化剤を更に含む、付記22に記載の方法。
（付記２５）
前記抗酸化剤が、トコフェロール、BHA、及びBHT、又はこれらの混合物から選択される、付記24に記載の方法。
（付記２６）
必要とする対象においてアポC-IIIを低減する方法であって、薬学的有効量の2-((5Z,8Z,11Z,14Z,17Z)-イコサ-5,8,11,14,17-ペンタエン-1-イルオキシ)ブタン酸:

又はその薬学的に許容される塩若しくはエステルを対象に投与することを含む、方法。
（付記２７）
2-((5Z,8Z,11Z,14Z,17Z)-イコサ-5,8,11,14,17-ペンタエン-1-イルオキシ)ブタン酸の薬学的有効量が、1用量当たり約200mg〜約800mgの範囲である、付記26に記載の方法。
（付記２８）
2-((5Z,8Z,11Z,14Z,17Z)-イコサ-5,8,11,14,17-ペンタエン-1-イルオキシ)ブタン酸を、1日1回投与する、付記27に記載の方法。
（付記２９）
薬学的有効量の式(I)の化合物

又はその薬学的に許容される塩若しくはエステルの使用であって、
式中、R₁及びR₂が、水素原子又は直鎖、分岐、及び/若しくは環式C₁〜C₆アルキル基から独立して選択され、但しR₁及びR₂が両方とも水素であることはなく、
必要とする対象においてアポリポタンパク質C-III(アポC-III)mRNA又はタンパク質を低減するための医薬の製造における、使用。
（付記３０）
前記化合物が、エナンチオマー、ジアステレオマー、又はこれらの混合物の形態で存在する、付記29に記載の使用。
（付記３１）
R₁及びR₂が、水素原子、メチル基、エチル基、n-プロピル基及びイソプロピル基から選択される、付記29に記載の使用。
（付記３２）
R₁及びR₂が、水素原子、メチル基、及びエチル基から選択される、付記29に記載の使用。
（付記３３）
R₁及びR₂の一方が水素原子であり、R₁及びR₂の他方はC₁〜C₃アルキル基から選択される、付記29に記載の使用。
（付記３４）
R₁及びR₂の一方が水素原子であり、R₁及びR₂の他方がメチル基又はエチル基から選択される、付記29に記載の使用。
（付記３５）
前記エステルが、グリセリド、及びC₁〜C₆アルキルエステルから選択される、付記29に記載の使用。
（付記３６）
前記エステルが、トリグリセリド、1,2-ジグリセリド、1,3-ジグリセリド、1-モノグリセリド、及び2-モノグリセリドから選択される、付記29に記載の使用。
（付記３７）
前記エステルが、メチルエステル、エチルエステル、イソプロピルエステル、n-ブチルエステル、及びtert-ブチルエステルから選択される、付記29に記載の使用。
（付記３８）
前記エステルが、メチルエステル及びエチルエステルから選択される、付記29に記載の使用。
（付記３９）
前記化合物が、そのR体で存在する、付記30に記載の使用。
（付記４０）
前記化合物が、そのS体で存在する、付記30に記載の使用。
（付記４１）
前記化合物が、ラセミ体で存在する、付記30に記載の使用。
（付記４２）
R₁が水素であり、R₂がエチルであり、前記式が

である、付記29に記載の使用。
（付記４３）
前記化合物が、式:

で表されるそのS及び/又はR体で存在する、付記42に記載の使用。
（付記４４）
式(I)の化合物の薬学的有効量が、1用量当たり約5mg〜約2gの範囲である、付記29に記載の使用。
（付記４５）
式(I)の化合物の薬学的有効量が、1用量当たり約200mg〜約800mgの範囲である、付記29に記載の使用。
（付記４６）
式(I)の化合物の薬学的有効量が、約600mgである、付記29に記載の使用。
（付記４７）
前記対象が、ヒトである、付記29に記載の使用。
（付記４８）
前記化合物を、1日1回投与する、付記29に記載の使用。
（付記４９）
前記化合物が、経口投与用の医薬組成物として製剤化される、付記29に記載の使用。
（付記５０）
前記医薬組成物が、ゼラチンカプセル剤又は錠剤の形態である、付記49に記載の使用。（付記５１）
前記医薬組成物が、少なくとも1種の結合剤、賦形剤、希釈剤、又はこれらの任意の組合せを更に含む、付記50に記載の使用。
（付記５２）
前記医薬組成物が、抗酸化剤を更に含む、付記50に記載の使用。
（付記５３）
前記抗酸化剤が、トコフェロール、BHA、及びBHT、又はこれらの混合物から選択される、付記52に記載の使用。
（付記５４）
薬学的有効量の2-((5Z,8Z,11Z,14Z,17Z)-イコサ-5,8,11,14,17-ペンタエン-1-イルオキシ)ブタン酸:

又はその薬学的に許容される塩若しくはエステルの使用であって、必要とする対象においてアポC-IIIを低減するための医薬の製造における、使用。
（付記５５）
2-((5Z,8Z,11Z,14Z,17Z)-イコサ-5,8,11,14,17-ペンタエン-1-イルオキシ)ブタン酸の薬学的有効量が、1用量当たり約200mg〜約800mgの範囲である、付記54に記載の使用。
（付記５６）
2-((5Z,8Z,11Z,14Z,17Z)-イコサ-5,8,11,14,17-ペンタエン-1-イルオキシ)ブタン酸を、1日1回投与する、付記55に記載の使用。
（付記５７）
前記対象が、スタチン療法中であり、約200mg/dl〜約499mg/dlのベースラインの空腹時トリグリセリドを有する、付記1に記載の方法。
（付記５８）
前記対象が、約200mg/dl〜約499mg/dlのベースラインの空腹時トリグリセリドを有する、付記1に記載の方法。
（付記５９）
アポC-IIIレベルが、少なくとも約20%低減される、付記57又は58に記載の方法。
（付記６０）
アポC-IIIレベルが、少なくとも約35%低減される、付記57又は58に記載の方法。
（付記６１）
前記対象が、スタチン療法中であり、500mg/dl超のベースラインの空腹時トリグリセリドを有する、付記1に記載の方法。
（付記６２）
前記対象が、500mg/dl超のベースラインの空腹時トリグリセリドを有する、付記1に記載の方法。
（付記６３）
アポC-IIIレベルが、少なくとも約25%低減される、付記61又は62に記載の方法。
（付記６４）
アポC-IIIレベルが、少なくとも約40%低減される、付記61又は62に記載の方法。
（付記６５）
前記対象が、少なくとも2.5mmol/L(約97mg/dl)の空腹時LDL-コレステロールを有する、付記1に記載の方法。
（付記６６）
アポC-IIIレベルが、少なくとも約25%低減される、付記65に記載の方法。
（付記６７）
アポC-IIIレベルが、少なくとも約40%低減される、付記65に記載の方法。
（付記６８）
必要とする対象においてアポC-IIIを低減する方法であって、薬学的有効量の脂質異常症薬剤及び式(I)の化合物:

又はその薬学的に許容される塩若しくはエステルを対象に投与することを含み、
式中、R₁及びR₂は、水素原子又は直鎖、分岐、及び/若しくは環式C₁〜C₆アルキル基から独立して選択され、但しR₁及びR₂が両方とも水素であることはない、方法。
（付記６９）
前記脂質異常症薬剤が、スタチンである、付記68に記載の方法。
（付記７０）
前記対象が、スタチン療法中であり、約200mg/dl〜約499mg/dlのベースラインの空腹時トリグリセリドを有する、付記29に記載の使用。
（付記７１）
前記対象が、約200mg/dl〜約499mg/dlのベースラインの空腹時トリグリセリドを有する、付記29に記載の使用。
（付記７２）
アポC-IIIレベルが、少なくとも約20%低減される、付記70又は71に記載の使用。
（付記７３）
アポC-IIIレベルが、少なくとも約35%低減される、付記70又は71に記載の使用。
（付記７４）
前記対象が、スタチン療法中であり、500mg/dl超のベースラインの空腹時トリグリセリドを有する、付記29に記載の使用。
（付記７５）
前記対象が、500mg/dl超のベースラインの空腹時トリグリセリドを有する、付記29に記載の使用。
（付記７６）
アポC-IIIレベルが、少なくとも約25%低減される、付記74及び75に記載の使用。
（付記７７）
アポC-IIIレベルが、少なくとも約40%低減される、付記74及び75に記載の使用。
（付記７８）
前記対象が、少なくとも2.5mmol/L(約97mg/dl)の空腹時LDL-コレステロールを有する、付記29に記載の使用。
（付記７９）
アポC-IIIレベルが、少なくとも約25%低減される、付記78に記載の使用。
（付記８０）
アポC-IIIレベルが、少なくとも約40%低減される、付記78に記載の使用。
（付記８１）
前記必要とする対象が、重度の高トリグリセリド血症、混合型脂質異常症、及び高コレステロール血症から選択される疾患又は状態を有する、付記1に記載の方法。
（付記８２）
前記必要とする対象が、重度の高トリグリセリド血症、混合型脂質異常症、及び高コレステロール血症から選択される疾患又は状態を有する、付記29に記載の使用。 The second difference factor is the required low dose of Compound A (600 mg QD) versus the 4 g dose in the EPA/DHA study. On a gram-to-gram basis, this difference was even greater for Compound A, clearly demonstrating the efficacy of Compound A molecules against EPA/DHA in reducing plasma apo C-III. As mentioned above, preclinical models suggest that apoC-III reduction is independent of TG reduction (Fig. 1).
(Appendix)
(Appendix 1)
A method of reducing apolipoprotein C-III (apo C-III) mRNA or protein in a subject in need thereof, wherein the method comprises a pharmaceutically effective amount of a compound of formula (I):

Or comprising administering to the subject a pharmaceutically acceptable salt or ester thereof,
In the formula, R ₁ and R ₂ are independently selected from a hydrogen atom or a linear, branched, and/or cyclic C ₁ -C ₆ alkyl group, provided that both R ₁ and R ₂ are hydrogen. Not the way.
(Appendix 2)
The method of claim 1 wherein the compound exists in the form of enantiomers, diastereomers, or mixtures thereof.
(Appendix 3)
The method according to Appendix 1, wherein R ₁ and R ₂ are selected from a hydrogen atom, a methyl group, an ethyl group, an n-propyl group and an isopropyl group.
(Appendix 4)
The method according to Appendix 1, wherein R ₁ and R ₂ are selected from a hydrogen atom, a methyl group, and an ethyl group.
(Appendix 5)
The method according to appendix 1, wherein _one of R ₁ and R ₂ is a hydrogen atom, and the other of R ₁ and R ₂ is selected from a C ₁ -C ₃ alkyl group.
(Appendix 6)
The method according to Appendix 1, wherein _one of R ₁ and R ₂ is a hydrogen atom, and the other of R ₁ and R ₂ is selected from a methyl group or an ethyl group.
(Appendix 7)
The ester is selected glycerides, and from C ₁ -C ₆ alkyl esters, method of statement 1.
(Appendix 8)
The method of claim 1 wherein the ester is selected from triglycerides, 1,2-diglycerides, 1,3-diglycerides, 1-monoglycerides, and 2-monoglycerides.
(Appendix 9)
The method of claim 1 wherein the ester is selected from methyl ester, ethyl ester, isopropyl ester, n-butyl ester, and tert-butyl ester.
(Appendix 10)
The method of claim 1 wherein the ester is selected from methyl ester and ethyl ester.
(Appendix 11)
The method of claim 2 wherein the compound is in its R configuration.
(Appendix 12)
The method of claim 2 wherein the compound is in its S configuration.
(Appendix 13)
The method of claim 2 wherein the compound exists as a racemate.
(Appendix 14)
R ₁ is hydrogen, R ₂ is ethyl and the above formula is

The method according to appendix 1.
(Appendix 15)
The compound has the formula:

15. The method according to appendix 14, which is present in its S and/or R form represented by
(Appendix 16)
The method of claim 1 wherein the pharmaceutically effective amount of the compound of formula (I) ranges from about 5 mg to about 2 g per dose.
(Appendix 17)
The method of claim 1 wherein the pharmaceutically effective amount of the compound of formula (I) ranges from about 200 mg to about 800 mg per dose.
(Appendix 18)
The method of claim 1 wherein the pharmaceutically effective amount of the compound of formula (I) is about 600 mg.
(Appendix 19)
The method according to Appendix 1, wherein the subject is a human.
(Appendix 20)
The method of Appendix 1, wherein the compound is administered once daily.
(Appendix 21)
The method of claim 1 wherein the compound is formulated as a pharmaceutical composition for oral administration. (Appendix 22)
22. The method according to appendix 21, wherein the pharmaceutical composition is in the form of a gelatin capsule or tablet. (Appendix 23)
23. The method of claim 22, wherein the pharmaceutical composition further comprises at least one binder, excipient, diluent, or any combination thereof.
(Appendix 24)
23. The method of claim 22 wherein the pharmaceutical composition further comprises an antioxidant.
(Appendix 25)
25. The method of appendix 24, wherein the antioxidant is selected from tocopherols, BHA, and BHT, or mixtures thereof.
(Appendix 26)
A method of reducing apo C-III in a subject in need thereof, comprising a pharmaceutically effective amount of 2-((5Z,8Z,11Z,14Z,17Z)-equosa-5,8,11,14,17-pentaene. -1-yloxy)butanoic acid:

Or administering a pharmaceutically acceptable salt or ester thereof to a subject.
(Appendix 27)
The pharmaceutically effective amount of 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaen-1-yloxy)butanoic acid is about 200 mg to about 800 mg per dose. The method according to appendix 26, which is in the range of.
(Appendix 28)
2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaen-1-yloxy)butanoic acid, once a day, the method according to Appendix 27 ..
(Appendix 29)
A pharmaceutically effective amount of a compound of formula (I)

Or the use of a pharmaceutically acceptable salt or ester thereof,
Wherein R ₁ and R ₂ are independently selected from a hydrogen atom or a linear, branched, and/or cyclic C ₁ -C ₆ alkyl group, provided that both R ₁ and R ₂ are hydrogen. Not,
Use in the manufacture of a medicament for reducing apolipoprotein C-III (apoC-III) mRNA or protein in a subject in need.
(Appendix 30)
Use according to claim 29, wherein the compound is present in the form of enantiomers, diastereomers, or mixtures thereof.
(Appendix 31)
The use according to appendix 29, wherein R ₁ and R ₂ are selected from a hydrogen atom, a methyl group, an ethyl group, an n-propyl group and an isopropyl group.
(Appendix 32)
The use according to appendix 29, wherein R ₁ and R ₂ are selected from a hydrogen atom, a methyl group, and an ethyl group.
(Appendix 33)
The use according to appendix 29, wherein _one of R ₁ and R ₂ is a hydrogen atom and the other of R ₁ and R ₂ is selected from C ₁ -C ₃ alkyl groups.
(Appendix 34)
The use according to appendix 29, wherein _one of R ₁ and R ₂ is a hydrogen atom and the other of R ₁ and R ₂ is selected from a methyl group or an ethyl group.
(Appendix 35)
The ester is selected glycerides, and from C ₁ -C ₆ alkyl esters, use of statement 29.
(Appendix 36)
Use according to claim 29, wherein the ester is selected from triglycerides, 1,2-diglycerides, 1,3-diglycerides, 1-monoglycerides, and 2-monoglycerides.
(Appendix 37)
Use according to claim 29, wherein the ester is selected from methyl ester, ethyl ester, isopropyl ester, n-butyl ester, and tert-butyl ester.
(Appendix 38)
Use according to claim 29, wherein the ester is selected from methyl ester and ethyl ester.
(Appendix 39)
31. The use according to appendix 30, wherein the compound is in its R configuration.
(Appendix 40)
31. The use according to appendix 30, wherein the compound is in its S configuration.
(Appendix 41)
31. The use according to appendix 30, wherein the compound exists in racemic form.
(Appendix 42)
R ₁ is hydrogen, R ₂ is ethyl and the above formula is

The use according to supplement 29.
(Appendix 43)
The compound has the formula:

The use according to appendix 42, which is present in its S and/or R form represented by
(Appendix 44)
Use according to claim 29, wherein the pharmaceutically effective amount of the compound of formula (I) ranges from about 5 mg to about 2 g per dose.
(Appendix 45)
Use according to claim 29, wherein the pharmaceutically effective amount of the compound of formula (I) ranges from about 200 mg to about 800 mg per dose.
(Appendix 46)
Use according to claim 29, wherein the pharmaceutically effective amount of the compound of formula (I) is about 600 mg.
(Appendix 47)
The use according to appendix 29, wherein the subject is a human.
(Appendix 48)
Use according to claim 29, wherein the compound is administered once daily.
(Appendix 49)
Use according to claim 29, wherein the compound is formulated as a pharmaceutical composition for oral administration.
(Appendix 50)
The use according to clause 49, wherein the pharmaceutical composition is in the form of a gelatin capsule or tablet. (Appendix 51)
The use according to clause 50, wherein the pharmaceutical composition further comprises at least one binder, excipient, diluent, or any combination thereof.
(Appendix 52)
The use according to clause 50, wherein the pharmaceutical composition further comprises an antioxidant.
(Appendix 53)
53. Use according to clause 52, wherein the antioxidant is selected from tocopherols, BHA, and BHT, or mixtures thereof.
(Appendix 54)
A pharmaceutically effective amount of 2-((5Z,8Z,11Z,14Z,17Z)-ikosa-5,8,11,14,17-pentaen-1-yloxy)butanoic acid:

Or the use of a pharmaceutically acceptable salt or ester thereof in the manufacture of a medicament for reducing apo C-III in a subject in need.
(Appendix 55)
The pharmaceutically effective amount of 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaen-1-yloxy)butanoic acid is about 200 mg to about 800 mg per dose. The use according to appendix 54, which is in the range of.
(Appendix 56)
2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaen-1-yloxy)butanoic acid administered once daily, use according to appendix 55 ..
(Appendix 57)
The method of claim 1, wherein the subject is on statin therapy and has a baseline fasting triglyceride of about 200 mg/dl to about 499 mg/dl.
(Appendix 58)
The method of claim 1, wherein the subject has a baseline fasting triglyceride of about 200 mg/dl to about 499 mg/dl.
(Appendix 59)
59. The method of clause 57 or 58, wherein the apo C-III level is reduced by at least about 20%.
(Appendix 60)
59. The method of clause 57 or 58, wherein the apo C-III level is reduced by at least about 35%.
(Appendix 61)
The method of claim 1 wherein the subject is on statin therapy and has a baseline fasting triglyceride greater than 500 mg/dl.
(Appendix 62)
The method of claim 1 wherein the subject has a baseline fasting triglyceride greater than 500 mg/dl.
(Appendix 63)
63. The method of clause 61 or 62, wherein the apo C-III level is reduced by at least about 25%.
(Appendix 64)
63. The method of clause 61 or 62, wherein the apo C-III level is reduced by at least about 40%.
(Appendix 65)
The method of claim 1 wherein the subject has a fasting LDL-cholesterol of at least 2.5 mmol/L (about 97 mg/dl).
(Appendix 66)
The method of claim 65, wherein the apo C-III level is reduced by at least about 25%.
(Appendix 67)
The method of claim 65, wherein the apo C-III level is reduced by at least about 40%.
(Appendix 68)
A method of reducing apo C-III in a subject in need thereof, comprising a pharmaceutically effective amount of a dyslipidemic agent and a compound of formula (I):

Or comprising administering to the subject a pharmaceutically acceptable salt or ester thereof,
In the formula, R ₁ and R ₂ are independently selected from a hydrogen atom or a linear, branched, and/or cyclic C ₁ -C ₆ alkyl group, provided that both R ₁ and R ₂ are hydrogen. Not the way.
(Appendix 69)
69. The method of appendix 68, wherein the dyslipidemic drug is a statin.
(Appendix 70)
Use according to claim 29, wherein the subject is on statin therapy and has a baseline fasting triglyceride of about 200 mg/dl to about 499 mg/dl.
(Appendix 71)
The use according to clause 29, wherein the subject has a baseline fasting triglyceride of about 200 mg/dl to about 499 mg/dl.
(Appendix 72)
The use according to clause 70 or 71, wherein the apo C-III level is reduced by at least about 20%.
(Appendix 73)
The use according to clause 70 or 71, wherein the apo C-III level is reduced by at least about 35%.
(Appendix 74)
Use according to claim 29, wherein the subject is on statin therapy and has a baseline fasting triglyceride of greater than 500 mg/dl.
(Appendix 75)
Use according to clause 29, wherein the subject has a baseline fasting triglyceride above 500 mg/dl.
(Appendix 76)
The use according to notes 74 and 75, wherein the apo C-III level is reduced by at least about 25%.
(Appendix 77)
The use according to notes 74 and 75, wherein the apo C-III level is reduced by at least about 40%.
(Appendix 78)
Use according to claim 29, wherein the subject has a fasting LDL-cholesterol of at least 2.5 mmol/L (about 97 mg/dl).
(Appendix 79)
The use according to note 78, wherein the apo C-III level is reduced by at least about 25%.
(Appendix 80)
The use according to clause 78, wherein the apo C-III level is reduced by at least about 40%.
(Appendix 81)
The method according to Appendix 1, wherein the subject in need has a disease or condition selected from severe hypertriglyceridemia, mixed dyslipidemia, and hypercholesterolemia.
(Appendix 82)
30. The use according to appendix 29, wherein the subject in need has a disease or condition selected from severe hypertriglyceridemia, mixed dyslipidemia, and hypercholesterolemia.

Claims (24)

A pharmaceutical composition for reducing apolipoprotein C-III (apo C-III) mRNA or protein in a subject in need thereof, wherein the pharmaceutical composition comprises a pharmaceutically effective amount of a compound of formula (I):

Or a pharmaceutically acceptable salt or ester thereof,
In the formula, R ₁ and R ₂ are independently selected from a hydrogen atom or a linear, branched, and/or cyclic C ₁ -C ₆ alkyl group, provided that both R ₁ and R ₂ are hydrogen. Not a pharmaceutical composition.   The pharmaceutical composition according to claim 1, wherein the compound is present in the form of an enantiomer, a diastereomer, or a mixture thereof. The pharmaceutical composition according to claim 1, wherein one of R ₁ and R ₂ is a hydrogen atom and the other of R ₁ and R ₂ is selected from a C ₁ -C ₃ alkyl group. The ester, glycerides, and are selected from C ₁ -C ₆ alkyl esters, pharmaceutical composition according to claim 1.   The pharmaceutical composition according to claim 1, wherein the ester is selected from methyl ester and ethyl ester. R ₁ is hydrogen, R ₂ is ethyl and the above formula is

The pharmaceutical composition according to claim 1, which is: The compound has the formula:

7. The pharmaceutical composition according to claim 6, which is present in its S and/or R form represented by   The pharmaceutical composition according to claim 1, wherein the pharmaceutically effective amount of the compound of formula (I) ranges from about 5 mg to about 2 g per dose.   The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition further comprises at least one binder, excipient, diluent, or any combination thereof.   The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition further comprises an antioxidant. A pharmaceutical composition for reducing apo C-III in a subject in need thereof, comprising a pharmaceutically effective amount of 2-((5Z,8Z,11Z,14Z,17Z)-equosa-5,8,11,14. ,17-Pentaen-1-yloxy)butanoic acid:

Or a pharmaceutical composition comprising a pharmaceutically acceptable salt or ester thereof. A pharmaceutically effective amount of a compound of formula (I)

Or the use of a pharmaceutically acceptable salt or ester thereof,
Wherein R ₁ and R ₂ are independently selected from a hydrogen atom or a linear, branched, and/or cyclic C ₁ -C ₆ alkyl group, provided that both R ₁ and R ₂ are hydrogen. Not,
Use in the manufacture of a medicament for reducing apolipoprotein C-III (apoC-III) mRNA or protein in a subject in need. R ₁ is hydrogen, R ₂ is ethyl and the above formula is

The use according to claim 12, which is The compound has the formula:

The use according to claim 13, which is present in its S and/or R form represented by:   13. The use according to claim 12, wherein the pharmaceutically effective amount of the compound of formula (I) ranges from about 200 mg to about 800 mg per dose.   2. The pharmaceutical composition of claim 1, wherein the subject is on statin therapy and has a baseline fasting triglyceride of about 200 mg/dl to about 499 mg/dl.   2. The pharmaceutical composition of claim 1, wherein the subject has a baseline fasting triglyceride of about 200 mg/dl to about 499 mg/dl.   2. The pharmaceutical composition of claim 1, wherein the subject is on statin therapy and has a baseline fasting triglyceride above 500 mg/dl.   2. The pharmaceutical composition of claim 1, wherein the subject has a baseline fasting triglyceride greater than 500 mg/dl. A pharmaceutical composition for reducing apo C-III in a subject in need thereof, comprising a pharmaceutically effective amount of a dyslipidemic agent and a compound of formula (I):

Or a pharmaceutically acceptable salt or ester thereof,
In the formula, R ₁ and R ₂ are independently selected from a hydrogen atom or a linear, branched, and/or cyclic C ₁ -C ₆ alkyl group, provided that both R ₁ and R ₂ are hydrogen. Not a pharmaceutical composition.   21. The pharmaceutical composition according to claim 20, wherein the dyslipidemic drug is a statin.   13. The use of claim 12, wherein the subject is on statin therapy and has a baseline fasting triglyceride of about 200 mg/dl to about 499 mg/dl.   13. The use of claim 12, wherein the subject has a baseline fasting triglyceride of about 200 mg/dl to about 499 mg/dl.   2. The pharmaceutical composition according to claim 1, wherein the subject in need has a disease or condition selected from severe hypertriglyceridemia, mixed dyslipidemia, and hypercholesterolemia.

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