WO2022045041A1 - Thioredoxin interacting protein expression inhibitor - Google Patents

Abstract

Disclosed is a thioredoxin interacting protein (TXNIP) expression inhibitor containing a curcumin derivative represented by formula (I) (In the formula, R1 are each independently a hydrogen atom, CH2F-, CHF2-, CF3-, CH2FO-, CHF2O-, or CF3O-, R2 are each independently a hydrogen atom, or a fluorine atom, A1 is a hydrogen atom, or methyl, A2 is alkyl, cyano, carboxy, alkoxycarbonyl, or R3-(CH2)m-, R3 is hydroxy, carboxy, cyano, alkylcarbonyloxy, alkoxycarbonyl, alkoxyalkoxy, hydroxyalkoxy, or CONR4R5, R4 and R5 are each independently a hydrogen atom, or alkyl, and m is an integer of 1-5), or a salt thereof.

Description

Thioredoxin interaction protein expression inhibitor

The present invention relates to a thioredoxin interaction protein expression inhibitor.

Thioredoxin interacting protein (hereinafter, also referred to as "TXNIP") binds to thioredoxin, which has an antioxidant effect, and inhibits its action. In humans, TXNIP increases with age and tolerance to oxidative stress decreases (Oberacker T et al. FEBS Letters 592: 2297-2307, 2018). And oxidative stress is associated with many diseases. Specific examples include sickle cell disease, atherosclerosis, Parkinson's disease, Alzheimer's disease, heart failure, myocardial infarction, schizophrenia, bipolar disorder, fragile X syndrome, and chronic fatigue syndrome.

In Drosophila, deficiency of TXNIP extends the lifespan (Oberacker T et al. FEBS Letters 592: 2297-2307, 2018).

In addition, TXNIP is deeply involved in the pathophysiology of cancer and diabetes. For example, in diabetes, hyperglycemic-induced TXNIP activates NLRP3 and triggers an inflammatory response (Zhou R et al. Nature Immunology 11: 136-141, 2010). TXNIP damages pancreatic beta cells. Knocking down TXNIP reduces beta cell damage and slows the progression of diabetes.

That is, reducing TXNIP leads to the treatment of various diseases related to oxidative stress, including diabetes. It has been reported in Patent Document 1 and Non-Patent Document 1 that such TXNIP is reduced.

In Patent Document 1, (a) the biological activity of the thioredoxin interacting protein (TXNIP), or (b) the gene encoding TXNIP, for applying a condition having a beneficial effect of improving resistance to oxidative stress to treatment. Compounds capable of reducing or suppressing expression have been reported. Examples of such compounds include antisense oligonucleotides or shRNAs that reduce or suppress the expression of the gene encoding TXNIP.

Non-Patent Document 1 reports that the antihypertensive agent verapamil lowers TXNIP levels and enhances the effect of insulin treatment in adult patients who have first developed type 1 diabetes.

In addition, the present inventors reported in Patent Document 2 that a curcumin derivative containing an F atom has high binding specificity for amyloid β protein and is useful as an active ingredient of a diagnostic imaging agent for Alzheimer's disease. ing.

Japan Special Table 2015-522251 Gazette International Publication No. 2010/098502

Nature Medicine 24: 1108-1112, 2018

An object of the present invention is to provide a TXNIP expression inhibitor having an excellent TXNIP expression inhibitory action, using a substance different from the conventional one as a novel active ingredient.

As a result of intensive studies to achieve the above object, the present inventors have found that the curcumin derivative described in Patent Document 2 described above strongly suppresses the expression of TXNIP at both the mRNA level and the protein level. .. However, curcumin did not show any inhibitory effect on the expression of TXNIP. Similarly, the compound in which the fluorine atom of the curcumin derivative described in Patent Document 2 was replaced with a hydrogen atom did not have the effect of suppressing the expression of TXNIP.

The present invention has been completed by further studies based on these findings, and provides the following TXNIP expression inhibitor.

Item 1. Equation (I):

（式中、R¹はそれぞれ独立にフッ素原子、CH₂F-、CHF₂-、CF₃-、CH₂FO-、CHF₂O-又はCF₃O-であり、R²はそれぞれ独立に水素原子又はフッ素原子であり、A¹は水素原子又はメチルであり、A²はアルキル、シアノ、カルボキシ、アルコキシカルボニル又はR³-(CH₂)_m-であり、R³はヒドロキシ、カルボキシ、シアノ、アルキルカルボニルオキシ、アルコキシカルボニル、アルコキシアルコキシ、ヒドロキシアルコキシ又はCONR⁴R⁵であり、R⁴及びR⁵はそれぞれ独立に水素原子又はアルキルであり、mは1～5の整数である）で表されるクルクミン誘導体又はその塩を含有する、チオレドキシン相互作用タンパク質(TXNIP)発現抑制剤。
項２．前記A¹が水素原子である、項１に記載のTXNIP発現抑制剤。
項３．前記A²がR³-(CH₂)_m-である、項１又は２に記載のTXNIP発現抑制剤。
項４．前記R³がカルボキシである、項３に記載のTXNIP発現抑制剤。

(In the equation, R ¹ is independently a fluorine atom, CH ₂ F-, CHF _2- , CF _3- , CH ₂ FO-, CHF ₂ O- or CF ₃ O-, and R ² is independently hydrogen. Atom or fluorine atom, A ¹ is hydrogen atom or methyl, A ² is alkyl, cyano, carboxy, alkoxycarbonyl or R ^3- (CH ₂ ) _m- , R ³ is hydroxy, carboxy, cyano, Alkoxycarbonyloxy, alkoxycarbonyl, alkoxyalkoxy, hydroxyalkoxy or CONR ⁴ R ⁵ , where R ⁴ and R ⁵ are independently hydrogen atoms or alkyl, where m is an integer from 1 to 5). A thioredoxin interacting protein (TXNIP) expression inhibitor containing a curcumin derivative or a salt thereof.
Item 2. Item 2. The TXNIP expression inhibitor according to Item 1, wherein A ¹ is a hydrogen atom.
Item 3. Item 2. The TXNIP expression inhibitor according to Item 1 or 2, wherein A ² is R ^3- (CH ₂ ) _m- .
Item 4. Item 3. The TXNIP expression inhibitor according to Item 3, wherein R ³ is carboxy.

The present invention also provides the following.
Item 5. A method for suppressing the expression of TXNIP, which comprises a step of administering a curcumin derivative represented by the above formula (I) or a salt thereof to a mammal in need thereof.
Item 6. Use of a curcumin derivative represented by the above formula (I) or a salt thereof in the production of a TXNIP expression inhibitor.
Item 7. Item 5. The method according to Item 5, or the use according to Item 6, wherein A ¹ is a hydrogen atom.
Item 8. Item 5. The method according to Item 5 or 7, or the use according to Item 6 or 7, wherein A ² is R ^3- (CH ₂ ) _m- .
Item 9. Item 5. The method according to Item 5, 7 or 8, wherein R ³ is carboxy, or the use according to Item 6, 7 or 8.

The curcumin derivative represented by the above formula (I) or a salt thereof has an excellent TXNIP expression inhibitory action, and is therefore useful as an active ingredient of the TXNIP expression inhibitor.

It is a graph which shows the analysis result of the expression suppression effect of TXNIP mRNA by curcumin, compound 1 and 3 carried out in Test Example 1. The vertical axis shows the amount of TXNIP mRNA (ratio when untreated is 1.0). Values are mean ± standard error, ^*** p <0.001, ^**** p <0.0001 vs DMSO, ns: not significant, n = 5-6 It is a graph which shows the analysis result of the expression suppression effect of the TXNIP protein by curcumin, compound 1 and 3 carried out in Test Example 1. The vertical axis shows the amount of TXNIP protein (ratio when untreated is 100). Values are mean ± standard error, ^*** p <0.001, ^**** p <0.0001 vs DMSO, ns: not significant, n = 5-6 It is a graph which shows the analysis result of the expression-suppressing action of the TXNIP mRNA of curcumin, compound 1 and 2 carried out in Test Example 2. The vertical axis shows the amount of TXNIP mRNA (ratio when untreated is 1.0). Values are mean ± standard error, ^** p <0.01, ^*** p <0.001, ^**** p <0.0001 vs control, ns: not significant, n = 3 It is a graph which shows the analysis result of the expression suppression effect of the TXNIP protein by curcumin, compound 1 and 2 carried out in Test Example 2. The vertical axis shows the amount of TXNIP protein (ratio when untreated is 100). Values are mean ± standard error, ^* p <0.05, ^** p <0.01, ^**** p <0.0001 vs control, ns: not significant, n = 3

Hereinafter, embodiments of the present invention will be described in detail.

In addition, in this specification, "comprise" also includes the meaning of "essentially consist of" and the meaning of "consist of".

The thioredoxin interaction protein (TXNIP) expression inhibitor of the present invention has the formula (I) :.

（式中、R¹はそれぞれ独立にフッ素原子、CH₂F-、CHF₂-、CF₃-、CH₂FO-、CHF₂O-又はCF₃O-であり、R²はそれぞれ独立に水素原子又はフッ素原子であり、A¹は水素原子又はメチルであり、A²はアルキル、シアノ、カルボキシ、アルコキシカルボニル又はR³-(CH₂)_m-であり、R³はヒドロキシ、カルボキシ、シアノ、アルキルカルボニルオキシ、アルコキシカルボニル、アルコキシアルコキシ、ヒドロキシアルコキシ又はCONR⁴R⁵であり、R⁴及びR⁵はそれぞれ独立に水素原子又はアルキルであり、mは1～5の整数である）で表されるクルクミン誘導体又はその塩を含有することを特徴とする。

(In the equation, R ¹ is independently a hydrogen atom, CH ₂ F-, CHF _2- , CF _3- , CH ₂ FO-, CHF ₂ O- or CF ₃ O-, and R ² is independently hydrogen. Atom or fluorine atom, A ¹ is hydrogen atom or methyl, A ² is alkyl, cyano, carboxy, alkoxycarbonyl or R ^3- (CH ₂ ) _m- , R ³ is hydroxy, carboxy, cyano, Alkoxycarbonyloxy, alkoxycarbonyl, alkoxyalkoxy, hydroxyalkoxy or CONR ⁴ R ⁵ , where R ⁴ and R ⁵ are each independently a hydrogen atom or alkyl, where m is an integer from 1 to 5). It is characterized by containing a curcumin derivative or a salt thereof.

The alkyl of A ² , R ⁴ and R ⁵ may be a straight-chain or branched C _1-6 alkyl, and a straight-chain or branched C _1-3 alkyl is preferable. The definition of alkyl also applies to the alkyl carbonyloxy, alkoxycarbonyl, alkoxyalkoxy and hydroxyalkoxy constituents of the curcumin derivative of formula (I).

Specific examples of C _1-6 alkyl include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, and hexyl.

Specific examples of C _1-3 alkyl include methyl, ethyl, n-propyl, and isopropyl.

It is desirable that the curcumin derivative of formula (I) does not cross the blood-brain barrier in order to prevent side effects on the brain, and in order to have such properties, A ² is R ^3- (CH ₂ ) _m- . Yes, it is preferred that R ³ is carboxy.

It is preferable that A ¹ is a hydrogen atom, that is, the substituent at the 4-position position of 1,6-heptadiene is only A ² .

When the curcumin derivative of the formula (I) or a salt thereof contains an asymmetric carbon, both the optical isomer and the racemate separated according to a conventional method are contained in the curcumin derivative of the present invention.

The curcumin derivative of the formula (I) may be a salt, and the salt may be any pharmaceutically acceptable salt, for example, an alkali metal salt such as a potassium salt or a sodium salt; a calcium salt. Alkaline earth metal salts such as; triethanolamine salts, organic amine salts such as tris (hydroxymethyl) aminomethane salts and the like. In addition, some of these salts have water of crystallization.

The curcumin derivative of the formula (I) in which A ¹ is a hydrogen atom can be produced according to a known method (for example, the method described in International Publication No. 2010/098502).

Further, the curcumin derivative of the formula (I) in which A ¹ is methyl or a salt thereof can be produced by the method described below.

The compound of formula (IA) can be produced by hydrolyzing the compound of formula (II).

（式中、R¹, R²及びA²は前述の通りである。）

(In the formula, R ¹ , R ² and A ² are as described above.)

Solvents for this reaction include alcohols such as methanol, ethanol, n-propanol, iso-propanol, butanol; ethers such as hydrous tetrahydrofuran and hydrodioxane; acid amides such as hydrous dimethylformamide and hydrodimethylacetamide; hydrous dimethyl. Examples thereof include sulfoxides such as sulfoxide and mixed solvents thereof.

It is desirable to add a mineral acid to promote this reaction, and examples of the mineral acid include hydrochloric acid, sulfuric acid, nitric acid, and perchloric acid. The mineral acid can be used in an amount of 3 to 10 times mol, preferably 4 to 6 times mol, with respect to the compound of formula (II).

This reaction can be carried out at 0 to 150 ° C, preferably 30 to 100 ° C, and the reaction time is usually about 1 to 150 hours.

The compound (II) can be produced by reacting the compound of the formula (III) with methyl iodide.

（式中、R¹, R²及びA²は前述の通りである。）

(In the formula, R ¹ , R ² and A ² are as described above.)

As the solvent for this reaction, aromatic hydrocarbons such as benzene, toluene and xylene; aliphatic hydrocarbons such as pentane, hexane, petroleum ether and ligroin; diethyl ether, dipropyl ether, dibutyl ether, tetrahydrofuran, dioxane and the like. Ethers; Ketones such as acetone and 2-butanone; nitriles such as acetonitrile and propionitrile; Acid amides such as dimethylformamide and dimethylacetamide; Sulfoxides such as dimethylsulfoxide and mixed solvents thereof. can.

In order to promote this reaction, it is desirable to add a base, which includes triethylamine, pyridine, N-methylmorpholin, 1,8-diazabicyclo [5,4,0] -7-undecene, N, N. -Organic bases such as dimethylaniline; alkali metal carbonates such as sodium carbonate and potassium carbonate; alkali metal hydrogen carbonates such as sodium hydrogen carbonate and potassium hydrogen carbonate; alkali metal water such as sodium hydroxide and potassium hydroxide Oxides: Hydroxides of alkaline earth metals such as barium hydroxide and calcium hydroxide can be mentioned. The base can be used in an amount of 3 to 20 times mol, preferably 5 to 10 times mol, with respect to the compound of formula (III).

This reaction can usually be carried out at 0 to 70 ° C, and the reaction time is about 1 to 48 hours.

In addition, it is desirable to use 2 to 20 times the molar amount of methyl iodide with respect to the compound of formula (III).

The compound of the formula (III) can be produced by condensing the compound of the formula (IV) and the compound of the formula (V). However, the compound of formula (IV) needs to undergo a 2-fold molar reaction with the compound of formula (V). The compound of the formula (V) can be produced by a known method.

（式中、R¹, R²及びA²は前述の通りである。）

(In the formula, R ¹ , R ² and A ² are as described above.)

In order to proceed the reaction efficiently, it is desirable to carry out the reaction in the presence of a boron compound and a base in a solvent. Examples of the boron compound that can be used in this reaction include boric acid, diboron trioxide, trimethyl borate, triethyl borate, tripropyl borate, tri-n-butyl borate, and tri-tert-butyl borate. Alternatively, a mixture of diboron trioxide and various boric acid esters can be mentioned. It is desirable to use the boric acid compound in an amount of 0.5 to 6 times the molar amount of the compound of the formula (IV).

Primary amines such as n-butylamine, sec-butylamine, tert-butylamine, n-propylamine, n-hexylamine, cyclohexylamine; morpholin, piperidine, 1,2,3,4-tetrahydro Secondary amines such as quinoline can be mentioned. It is desirable to use 1 times the molar amount of the base with respect to the compound of the formula (V).

As the solvent, aromatic hydrocarbons such as benzene, toluene and xylene; aliphatic hydrocarbons such as pentane, hexane, heptane, petroleum ether and ligroin; diethyl ether, dipropyl ether, dibutyl ether, tetrahydrofuran and dioxane. Ethers such as; esters such as methyl acetate, ethyl acetate, methyl propionate; acid amides such as dimethylformamide and dimethylacetamide; sulfoxides such as dimethylsulfoxide; phosphate amides such as hexamethylphosphortriamide and these. Can be mentioned.

The reaction temperature can be usually 0 to 150 ° C, preferably 0 to 100 ° C, and the reaction time is usually about 0.5 to 24 hours.

Further, in the above reaction, after the reaction, it is necessary to treat the reaction solution with an acid in order to decompose the boron complex of the compound of the formula (III) produced. Examples of the acid used at that time include mineral acids such as hydrochloric acid and sulfuric acid, and organic acids such as acetic acid and propionic acid.

The compound of formula (IV) can be produced by reacting the compound of formula (VI) with chlorodimethyl ether. The compound of the formula (VI) can be produced by a known method.

（式中、R¹及びR²は前述の通りである。）

(In the formula, R ¹ and R ² are as described above.)

Solvents include aromatic hydrocarbons such as benzene, toluene and xylene; aliphatic hydrocarbons such as pentane, hexane, heptane, petroleum ether and ligroin; diethyl ether, dipropyl ether, dibutyl ether, tetrahydrofuran, dioxane and the like. Ethers; Ketones such as acetone and 2-butanone; nitriles such as acetonitrile and propionitrile; Acid amides such as dimethylformamide and dimethylacetamide; Sulfoxides such as dimethylsulfoxide; dichloromethane, carbon tetrachloride, 1, 2 -Halogenized hydrocarbons such as dichloroethane and mixed solvents thereof can be mentioned.

It is desirable to add a base to promote this reaction, and the bases include triethylamine, pyridine, N-methylmorpholin, N-methylpiperidine, 1,8-diazabicyclo [5,4,0] -7. -Organic bases such as undecene, N, N-dimethylaniline; alkali metal carbonates such as sodium carbonate and potassium carbonate; alkali metal hydrogen carbonates such as sodium hydrogen carbonate and potassium hydrogen carbonate; sodium hydroxide and potassium hydroxide Alkali metal hydroxides and the like can be mentioned. The base can be used in an amount of 1-3 times mol, preferably 1.4-1.8 times mol, of the compound of formula (VI).

This reaction can usually be carried out at 0 to 50 ° C, and the reaction time is usually about 1 to 48 hours.

The curcumin derivative of the formula (I) obtained by the above-mentioned production method and the method incidental thereto can be obtained by known means, for example, concentration, vacuum concentration, distillation, fractional distillation, transsolution, solvent extraction, crystallization, recrystallization, chromatography. It can be isolated and purified by imaging or the like.

When the curcumin derivative of the formula (I) is obtained in a free form, a salt can be formed by a usual method.

Specific examples of the curcumin derivative of the formula (I) are shown in Table 1 below.

Most of the curcumin derivatives of formula (I) are hydrophobic compounds and have low solubility in water. It is desirable that the compound to be administered to a living body has high water solubility, and among the curcumin derivatives of the formula (I), those having a salt are more preferable.

Since the curcumin derivative of the formula (I) or a salt thereof has an excellent TXNIP expression inhibitory action at both the mRNA level and the protein level, it can be used as an active ingredient of the TXNIP expression inhibitor.

The base sequence of the TXNIP gene is registered as RefSeq Accession No. NM_006472 (human), NM_001313972 (human), etc. on the NCBI website, and the amino acid sequence is RefSeq Accession No. NP_006463 (human), NP_001300901 (human). It is registered as such.

In addition, suppression of TXNIP expression means reducing the expression of the TXNIP gene, which can be confirmed by a decrease in the amount of transcript produced from the TXNIP gene, a decrease in the amount of translated product produced from the TXNIP gene, etc. can.

By suppressing the expression of TXNIP, resistance to oxidative stress is improved. Diseases related to oxidative stress include diabetes, atherosclerosis, Parkinson's disease, Alzheimer's disease, arteriosclerosis, stroke, heart failure, myocardial infarction, schizophrenia, bipolar disorder, sickle erythema, fragile X syndrome, Chronic fatigue syndrome, cancer (eg gastric cancer, colon cancer (rectal cancer, colon cancer), small bowel cancer, liver cancer, pancreatic cancer, lung cancer, pharyngeal cancer, esophageal cancer, renal cancer, bile and bile duct cancer, head and neck cancer, Bladder cancer, prostate cancer, breast cancer, uterine cancer (cervical cancer, uterine body cancer), ovarian cancer, brain tumor, thoracic adenoma, leukemia, malignant lymphoma, etc.) can be mentioned. It can be used for the treatment and prevention of cancer.

The curcumin derivative of formula (I) is highly safe because its basic skeleton is curcumin, which is a food ingredient.

The TXNIP expression inhibitor of the present invention can be used as a pharmaceutical composition, a food composition, or the like.

The pharmaceutical composition of the present invention is administered to mammals including humans. Administration of the pharmaceutical composition of the present invention may be topical or systemic. The administration method is not particularly limited, and it is administered orally or parenterally. Parenteral routes of administration include subcutaneous, intraperitoneal, intravenous, arterial or spinal fluid injections or infusions, percutaneous administration and the like.

The pharmaceutical composition of the present invention contains a pharmaceutically acceptable form suitable for administration to humans and a physiologically acceptable additive. Such pharmaceutical compositions are optionally pharmaceutically acceptable diluents, buffers, solubilizers (eg, cyclodextrin, polyethylene glycol, or Tween ™, Pluronic ™, Cremofol ™, phospholipids). Surfactants such as), soothing agents and the like may be added, and if necessary, they contain components such as pharmaceutically acceptable solvents, stabilizers or antioxidants (eg ascorbic acid). But it may be. The dose of the pharmaceutical composition of the present invention is appropriately selected depending on the usage, the age of the patient, the sex and other conditions, and the degree of the disease.

The content of the curcumin derivative of the formula (I) or a salt thereof in the pharmaceutical composition of the present invention can be appropriately selected from the range of 0.01 to 100% by mass, preferably 0.1 to 100% by mass.

The food composition of the present invention includes any food composition that can be ingested by animals (including humans). The food composition of the present invention may contain amino acids, nucleic acids, minerals, vitamins, flavonoids, quinones, polyphenols, binders, refreshing agents, sweeteners, essential fatty acids, disintegrants, slippers, as required. Agents, fragrances, stabilizers, colorants, preservatives, surfactants, sustained release adjusters, solubilizers, wetting agents and the like can be blended.

The type of the food composition of the present invention is not particularly limited, and for example, beverages (soft beverages such as coffee, juice, and tea beverages, dairy beverages, carbonated beverages, Japanese liquor, western liquor, liquors such as fruit liquor, etc.). ; Spreads (custard cream, etc.); Pastes (fruit paste, etc.); Western confectionery (chocolate, donuts, pie, cream puff, gum, jelly, candy, cookies, cakes, pudding, etc.); Japanese confectionery (Daifuku, rice cake, bun) , Castella, Anmitsu, Sheep, etc.); Iced drinks (ice cream, ice drinks, sherbets, etc.); Foods (curry, beef bowl, miscellaneous dishes, miso juice, soup, meat sauce, pasta, pickles, jam, royal jelly, etc.); ; Fermented foods (yogurt, royal jelly, etc.); Seasonings (dressing, sprinkle, delicious seasonings, soup base, etc.) and the like.

The food composition of the present invention can also be used as a health food, a functional food, a dietary supplement, a supplement, a food for specified health use, or a food with a functional claim. The dosage unit form when used as a supplement is not particularly limited and may be appropriately selected, and examples thereof include tablets, granules, liquids, capsules, and powders.

The content of the curcumin derivative of the formula (I) or a salt thereof in the food composition of the present invention can be appropriately selected from the range of 0.01 to 100% by mass, preferably 0.1 to 100% by mass in the total amount of the food composition. ..

The intake amount of the food composition of the present invention can be appropriately set according to various conditions such as the weight, age, sex, and symptom of the ingestor.

Next, synthetic examples and test examples related to the present invention will be described, but the present invention is not limited thereto. The NMR spectra in the following synthetic examples were measured using JEOL RESONANCE ECZ-400S.

[Synthesis Example 1] Synthesis of 1,7-bis (4'-hydroxy-3'-trifluoromethoxy) phenyl-4-carboxypropyl-1,6-heptadiene-3,5-dione (Compound 2) International Publication 1,7-Bis (4'-hydroxy-3'-trifluoromethoxy) phenyl-4-methoxycarbonylpropyl-1,6-heptadiene-3,5-dione synthesized with reference to the description of 2010/098502 90 mg (0.156 mmol) was added to 4.7 mL (0.468 mmol) of 0.1 M-sodium hydroxide aqueous solution, and the mixture was stirred at room temperature for 2 hours. After adjusting the pH to 2 by adding 1M-hydrochloric acid to the reaction solution, the mixture was extracted with ethyl acetate. The extract was washed with water and saturated brine, dried over magnesium sulfate, and the solvent was distilled off under reduced pressure. When the obtained residue is purified by silica gel column chromatography (eluent: ethyl acetate: hexane = 1: 1), 1,7-bis (4'-hydroxy-3'-trifluoromethoxy) phenyl at a melting point of 160-161 ° C. -4-carboxypropyl-1,6-heptadiene-3,5-dione 35 mg (40.0%) was obtained.
¹⁹ FNMR (d ₆ DMSO): δ -58.34 (s), -58.47 (s), ¹ HNMR (d ₆ DMSO): δ1.49 (0.7H, m), δ1.63 (1.3H, m), δ1 .84 (0.7H, m), δ2.24 (0.7H, m), δ2.33 (0.5H, m), δ2.49 (0.7H, m), δ2.5-2.7 (1.5H), δ4 .58 (0.3H, t, J = 7.0Hz), δ6.95 (0.7H, d, J = 15.6Hz), δ7.04 (0.7H, d, J = 9.0Hz), δ7.06 (1.3H) , d, J = 9.0Hz), δ7.28 (1.3H, d, J = 15.6Hz), δ7.61 (0.7H, dd, J = 2.0Hz, 9.0Hz), δ7.63 (1.3H, d , J = 15.6Hz), δ7.64 (0.7H, d, J = 15.6Hz), δ7.65 (0.7H, br.s), δ7.75 (1.3H, dd, J = 2.0Hz, 9.0Hz ), δ7.78 (1.3H, br.s), δ17.80 (0.7H, s).

[Synthesis Example 2] Synthesis of 1,7-bis (4'-hydroxy-3'-methoxy) phenyl-4-carboxyethyl-1,6-heptadiene-3,5-dione (Compound 3) (1) 4- A solution of methyl acetyl-5-oxohexanoate 186 mg (1.0 mmol) (Sigma-Aldrich) and diboron trioxide 56 mg (0.8 mmol) (Nakalitesk Co., Ltd.) in ethyl acetate (2 mL) at 60 ° C for 30 minutes. After heating, add 304 mg (2.0 mmol) of vanillin (Nakaraitesk Co., Ltd.) and 0.54 mL (2.0 mmol) of tri-n-butyl borate (Tokyo Kasei Kogyo Co., Ltd.), and heat at the same temperature for another 30 minutes. Continued. Then, 0.1 mL (1.0 mmol) of n-butylamine (Nacalai Tesque, Inc.) was added, and the mixture was heated at the same temperature for 4 hours. After cooling the reaction solution to room temperature, 1M-hydrochloric acid (2 mL) was added, and the mixture was vigorously stirred for 15 minutes. The reaction mixture was extracted with ethyl acetate, the extract was washed with water, then washed with saturated brine, and dried over magnesium sulfate. When the solvent was distilled off under reduced pressure and the obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate: hexane = 1: 1), a small amount of dichloromethane was added to the obtained substance and left at room temperature. , 7,7-Bis (4'-hydroxy-3'-methoxy) phenyl-4-methoxycarbonylethyl-1,6-heptadiene-3,5-dione 208 mg (45.8%) having a melting point of 124-125 ° C. Was done.
¹ HNMR (d ₆ DMSO): δ2.07 (1.3H, m), δ2.31 (1.3H, m), δ2.48 (0.7H, m), δ2.98 (0.7H, m), δ3. 54 (1H, s), δ3.59 (2H, s), δ3.80 (4H, s), δ3.85 (2H, s), δ4.58 (0.6H, m), δ6.80 (1.3H) , d, J = 8.0Hz), δ6.83 (0.7H, d, J = 8.0Hz), δ6.91 (1.3H, d, J = 15.6Hz), δ7.13 (0.7H, d, J = 15.6Hz), δ7.16 (1.3H, dd, J = 8.0Hz, 2.0Hz), δ7.23 (0.7H, dd, J = 8.0Hz, 2.0Hz), δ7.32 (1.3H, d, J) = 2.0Hz), δ7.35 (0.7H, d, J = 2.0Hz), δ7.60 (0.7H, d, J = 15.6Hz), δ7.61 (1.3H, d, J = 15.6Hz), δ18.03 (0.4H, s).

(2) 1,7-Bis (4'-hydroxy-3'-methoxy) phenyl-4-methoxycarbonylethyl-1,6-heptadiene-3,5-dione 182 mg obtained in the above step (1) (2) 0.4 mmol) was added to 12 mL (1.2 mmol) of a 0.1 M-sodium hydroxide aqueous solution, and the mixture was stirred at room temperature for 1 hour. After adjusting the pH to 2 by adding 1M-hydrochloric acid to the reaction solution, the mixture was extracted with ethyl acetate. The extract was washed with water and saturated brine, dried over magnesium sulfate, and the solvent was distilled off under reduced pressure. When the obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate), it was 1,7-bis (4'-hydroxy-3'-methoxy) phenyl-4-carboxyethyl-1 having a melting point of 150-151 ° C. , 6-Heptadien-3,5-dione 126 mg (71.5%) was obtained.
¹ HNMR (d ₆ DMSO): δ2.04 (1.3H, m), δ2.22 (1.3H, m), δ2.40 (0.7H, m), δ2.93 (0.7H, m), δ3. 79 (4H, s), δ3.83 (2H, s), δ4.56 (0.6H, m), δ6.79 (0.7H, d, J = 8.0Hz), δ6.81 (0.7H, d, J = 15.6Hz), δ6.90 (1.3H, d, J = 15.6Hz), δ7.1-7.25 (3.3H), δ7.31 (1.3H, d, J = 2.0Hz), δ7.33 ( 0.7H, d, J = 2.0Hz), δ7.59 (0.7H, d, J = 15.6Hz), δ7.60 (1.3H, d, J = 15.6Hz), δ18.00 (0.4H, s) ..

Compound 1 used in the following test examples was synthesized according to the description in Synthesis Example 2 of International Publication No. 2010/098502. Compound 1 has the following structure.

[Test Example 1] Analysis of TXNIP expression inhibitory effect of compounds 1 and 3 ARPE-19 cells were seeded in DMEM / F-12 (FBS 10%) medium in a 12-well plate at an amount of 2 × 10 ⁵ cells / well. .. Then, it was adhered for 24 hours. The cells were then treated with medium containing or not containing curcumin, compound 1 or compound 3 (1 μM or 5 μM) for 24 hours. After treatment, cells were extracted for RNA or protein analysis. Then, the amount of TXNIP mRNA was analyzed using real-time PCR, and the amount of TXNIP protein was analyzed using Western blotting.

The obtained results are shown in FIGS. 1 and 2. The amount of TXNIP mRNA (Fig. 1) was significantly lower in cells treated with compound 1 at 1 μM and 5 μM concentrations than in untreated cells, but untreated in cells treated with curcumin and compound 3 at 1 μM and 5 μM concentrations. No difference from the cells was observed. Similarly, the amount of TXNIP protein (Fig. 2) was significantly lower in cells treated with Compound 1 at 1 μM and 5 μM concentrations than in untreated cells, but in cells treated with curcumin and Compound 3 at 1 μM and 5 μM concentrations. No difference from untreated cells was observed. From the above results, it was suggested that Compound 1 has an inhibitory effect on the expression of TXNIP.

[Test Example 2] Analysis of TXNIP expression inhibitory effect of compounds 1 and 2 ARPE-19 cells were seeded in DMEM / F-12 (FBS 10%) medium in a 12-well plate at an amount of 2 × 10 ⁵ cells / well. .. Then, it was adhered for 24 hours. The cells were then treated with medium containing or not containing curcumin, compound 1 or compound 2 (0.5 μM, 1 μM or 3 μM) for 24 hours. After treatment, cells were extracted for RNA or protein analysis. Then, the amount of TXNIP mRNA was analyzed using real-time PCR, and the amount of TXNIP protein was analyzed using Western blotting.

The obtained results are shown in FIGS. 3 and 4. In FIG. 3, a concentration-dependent decrease in the amount of TXNIP mRNA was observed in cells treated with compounds 1 and 2 at concentrations of 0.5 μM, 1 μM, or 3 μM, but differences from untreated cells in cells treated with curcumin. Was not recognized. Similarly, the amount of TXNIP protein (Fig. 4) was decreased in the cells treated with Compound 1 and Compound 2 in a concentration-dependent manner, but was different from that in the untreated cells in the cells treated with curcumin. I couldn't. From the above results, it was suggested that not only compound 1 but also compound 2 has a TXNIP expression inhibitory effect.

Claims (4)

Equation (I):

(In the equation, R ¹ is independently a fluorine atom, CH ₂ F-, CHF _2- , CF _3- , CH ₂ FO-, CHF ₂ O- or CF ₃ O-, and R ² is independently hydrogen. Atom or fluorine atom, A ¹ is hydrogen atom or methyl, A ² is alkyl, cyano, carboxy, alkoxycarbonyl or R ^3- (CH ₂ ) _m- , R ³ is hydroxy, carboxy, cyano, Alkoxycarbonyloxy, alkoxycarbonyl, alkoxyalkoxy, hydroxyalkoxy or CONR ⁴ R ⁵ , where R ⁴ and R ⁵ are independently hydrogen atoms or alkyl, where m is an integer from 1 to 5). A thioredoxin interacting protein (TXNIP) expression inhibitor containing a curcumin derivative or a salt thereof. The TXNIP expression inhibitor according to claim 1, wherein A ¹ is a hydrogen atom. The TXNIP expression inhibitor according to claim 1 or 2, wherein A ² is R ^3- (CH ₂ ) _m- . The TXNIP expression inhibitor according to claim 3, wherein R ³ is carboxy.

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