KR100843319B1 - GAMMA-Lactone compound as a PPARDELTA; Ligand and pharmaceutical, cosmetic and food compositions thereof - Google Patents

Abstract

The present invention provides a pharmaceutical composition of gamma lactone (γ-lactone) represented by the following general formula (I) having the activity on the Peroxysome Proliferator Activated Receptor δ (hereinafter referred to as 'PPARδ') The present invention relates to a pharmaceutical composition, a functional food, a functional cosmetic, and an animal feed composition having an acceptable salt as an active ingredient.

[Formula I]

Marine natural products, peroxisomal multipliers, activation, receptors, obesity, diabetes, cosmetics, muscle enhancers, functional foods, animal feed compositions

Description

Γ-Lactone compound as a PPARδ ligand and pharmaceutical, cosmetic and food compositions comprising

The present invention provides a gamma lactone compound represented by the compound of formula (I) below, which is a Peroxysome Proliferator Activated Receptor δ (PPARδ) activating ligand that can be used for the treatment of obesity, hyperlipidemia, arteriosclerosis and diabetes or A pharmaceutical composition, a functional food, a functional cosmetic, and an animal feed composition comprising the pharmaceutically acceptable salt of the compound as an active ingredient.

[Formula I]

Among the nuclear receptors, the Peroxysome Proliferator Activated Receptor (PPAR) has three known subtypes: PPARα, PPARγ, and PPARδ ( Nature , 1990 , 347 , p645-650., Proc. Natl. Acad. Sci . USA 1994 , 91 , p7335-7359). PPARα, PPARγ and PPARδ have distinct functions according to tissues in vivo, and expression sites also differ. PPARα is mainly expressed in human heart, kidney, skeletal muscle, and large intestine ( Mol . Pharmacol . 1998 , 53 , p14-22., Toxicol . Lett . 1999 , 110 , p119-127., J. Biol . Chem . 1998 , 273 , p16710-16714), and the β-oxidation of peroxisome and mitochondria ( Biol . Cell. 1993 , 77 , p67-76., J. Biol . Chem . 1997 , 272 , p27307-27312 ). PPARγ is weakly expressed in skeletal muscle, but is expressed in large amounts in adipose tissue, and is known to be involved in the differentiation and storage of energy in fat form and the regulation of insulin and sugar homeostasis ( Moll . Cell . 1999 , 4 , p585). -594., P 597-609., P611-617). PPARδ is evolutionarily conserved in mammals, including humans, and vertebrates such as rodents and sea lions. Studies so far PPARδ is known to play an important role in the process of expression and germ cell (Genes Dev . 1999 , 13 , p1561-1574.), Differentiation of neurons in the Central Nervous System (CNS) ( J. Chem . Neuroanat 2000 , 19 , p225-232), wound healing through anti-inflammatory effects ( Genes Dev . 2001 , 15 , p3263-3277., Proc . Natl . Acad . Sci . USA 2003 , 100 , p6295-6296). Recent studies have demonstrated that PPARδ is involved in adipocyte differentiation and fat metabolism ( Proc . Natl . Acad . Sci . USA 2002 , 99 , p303-308., Mol . Cell . Biol . 2000 , 20 , p5119 It was found that PPARδ activates the expression of uncoupling proteins (UCPs), a key gene involved in β-oxidation and a gene involved in energy metabolism during fatty acid degradation ( Nature 2000 , 406 , p415-418., Cell 2003 , 113 , p159-170., PLoS Biology 2004 , 2 , p1532-1539). Activation of PPARδ also increases HDL, improves type 2 diabetes in the absence of weight change ( Proc . Natl . Acad . Sci. USA 2001 , 98 , p5306-5311., 2003 , 100 , p15924-15929), Atherosclerosis treatment is also possible by suppressing atherosclerotic disease related genes ( Science , 2003 , 302 , p453-457). PPARδ also affects mitochondrial production and muscle fiber transformation in muscle by PGC-1α. Muscle muscle fibers include fatty acid-degrading muscle fibers (Type I) that promote endurance and glycolytic muscle fibers (Type II) that promote quickness. Fatty acid-degrading muscle fibers (Type I), which promote endurance, are red because they contain a lot of mitochondria and myoglobin, while glycolytic muscle fibers (Type II), which promote quickness, are relatively white. As a result of overexpression of PGC-1α artificially in the muscles of the mice, it was confirmed that a significant portion of the muscles were changed from Type II to Type I muscle containing a lot of mitochondria. This can be interpreted as a result of the production of mitochondria by PGC-1α. It has been reported that PPARδ regulates β-oxidized UCP expression, mitochondrial production and transformation of muscle fibers of fatty acids. (PLoS Biology, 2004 , 2: e294). Therefore, the regulation of fat metabolism using PPARδ provides important clues for treating obesity, diabetes, hyperlipidemia and arteriosclerosis, and for solving muscle weakness through muscle fiber transformation.

Gamma lactone of Formula 1 is a sponge animal Plakortis It has been isolated from nigra and reported cytotoxicity against HCT-cancer cells ( J. Nat . Prod . 2002, 65, 1258), but its physiological activity against peroxysomal proliferative activator receptor δ is unknown.

An object of the present invention is a gamma lactone compound represented by the formula (1) or a pharmaceutically acceptable compound of the peroxysomal proliferative activating receptor δ activating ligand for the prevention or treatment of obesity, hyperlipidemia, arteriosclerosis and diabetes, and muscle strengthening It is to provide a pharmaceutical, functional cosmetics, functional foods and animal feed compositions comprising the salt as an active ingredient.

The present invention relates to a gamma lactone compound represented by the compound of formula (I) below or a pharmaceutically acceptable compound of the present invention, which is a peroxysomal proliferative activating receptor δ activating ligand that can be used for the treatment of obesity, hyperlipidemia, arteriosclerosis and diabetes, and muscle strengthening It is to provide a pharmaceutical, functional cosmetics, functional food and a composition for animal feed having an acceptable salt as an active ingredient.

[Formula I]

The gamma lactone of the formula (I) according to the present invention can be prepared through the following reaction scheme.

[In the above scheme, R is an alkyl group, aryl group, alkaryl group having 1 to 20 carbon atoms, the alkyl group, aryl group and alkylaryl group may further include a nitrogen atom.]

[Step 1] Preparation of Compound Represented by Formula (IV)

This step is a step of synthesizing the alkylmethyl ketone represented by the formula (IV) through 1,4-addition reaction to the vinyl menyl ketone represented by the formula (III).

The compound represented by the formula (IV) is obtained by reacting the alkyl bromide represented by the formula (II) with nickel (II) chloride under a zinc catalyst. The amount of nickel (II) chloride to be used may be used in an amount of 1 to 5 equivalents, preferably 1 to 2 equivalents, and 5 to 10 equivalents of zinc, preferably 5 equivalents. As the solvent to be used, ethers such as tetrahydrofuran and diethyl ether and amines such as pyridine, triethylamine and diethylamine may be used. Preferably, a single tetrahydrofuran solvent, a pyridine single solvent or tetra Preference is given to a mixed solvent of hydrofuran and pyridine.

The reaction temperature is carried out at 0 ~ 100 ℃ but preferably 50 ~ 60 ℃.

[Step 2] Preparation of Compound Represented by Formula (I)

A compound represented by the formula (I) is obtained by reacting a compound represented by the formula (IV) with a compound represented by the formula (V) under a samarium iodide (SmI ₂ ) catalyst. The reaction is possible at -78 ° C to room temperature, but 0-10 ° C is preferred. As the solvent to be used, ethers such as tetrahydrofuran and diethyl ether may be used, but a tetrahydrofuran single solvent is preferable.

The gamma lactone compound represented by the general formula (I) thus obtained is an important substance as a ligand of the PPAR type protein. This compound also has two chiral carbons, so four stereoisomers are present. The scope of the present invention includes gamma lactone compounds represented by the formula (I), stereoisomers thereof, each enantiomer thereof and salts thereof.

The present invention is an obesity, diabetes mellitus, hyperlipidemia, hypercholesterolemia, cyclopoproteins, arteries using the gamma lactone compound represented by the formula (I) or a pharmaceutically acceptable salt of the compound as an active ingredient Provided is a pharmaceutical composition for the prevention and treatment of sclerosis, hypertension, circulatory disease, ischemic heart disease and muscle strengthening.

[Formula I]

Gamma lactone (γ-lactone) compound represented by the formula (I) is characterized by having the following structure.

In addition, the present invention is an obesity, diabetes mellitus, hyperlipidemia, hypercholesterolemia, hyperlipoproteinosis, using the gamma lactone compound represented by the formula (I) or a pharmaceutically acceptable salt of the compound as an active ingredient Provide functional foods for preventing atherosclerosis, hypertension, circulatory disease, and ischemic heart disease.

In another aspect, the present invention provides a functional cosmetic for preventing obesity and improving obesity as a gamma lactone compound represented by the formula (I) or a pharmaceutically acceptable salt of the compound as an active ingredient.

In another aspect, the present invention provides a composition for animal feed comprising a gamma lactone compound represented by the formula (I) or a pharmaceutically acceptable salt of the compound as an active ingredient.

In another aspect, the present invention is a peroxysomal proliferator activated receptor δ using a gamma lactone compound represented by the formula (I) or a pharmaceutically acceptable salt of the compound as an active ingredient An activator composition is provided.

EXAMPLE

Hereinafter, the method of the present invention will be described in detail with reference to Examples. However, the present invention is not limited to these examples.

Example 1 Preparation of a Compound Represented by Formula (IV) (Step 1)

879 mg (3.7 mmol) of nickel (II) chloride, 1.1 g (16.8 mmol) of Zn dust, 1.37 ml (16.8 mmol) of 3-buten-2-one of the formula III, 50 ml of pyridine Into and stirred at 60 ° C. for 30 minutes. 1 g (3.4 mmol) of 1- (10-bromodecyl) benzene of Formula II was dissolved in 5 ml of pyridine, and this was added to the reaction solution. The reaction solution was stirred at 60 ° C. for 12 hours, filtered through celite to remove solid suspension, and pyridine was removed in a vacuum evaporator. 2N HCl and ethyl acetate were added to the concentrated residue, the organic layer was separated, dried over magnesium sulfate, filtered and concentrated. The concentrated residue was separated by silica gel column chromatography (hexane: ethyl acetate = 10: 1) to give 470 mg of 14-phenyltetradecan-2-one of formula IV (yield: 48.4%). Got.

¹ H NMR (300 MHz, CDCl ₃ ): δ 7.13-7.29 (m, 5H), 2.59 (t, 2H, J = 7.5 Hz), 2.40 (t, 2H, J = 7.4 Hz), 2.12 (s, 3H ), 1.60 (m, 4H), 1.25 (m, 16H) ¹³ C NMR (75.5 MHz, CDCl ₃ ): δ 209.5, 143.1, 128.6, 128.4, 125.7, 44.0, 36.2, 31.7, 30.0, 29.8, 29.76, 29.70 , 29.6, 29.59, 29.52, 29.4, 24.0

Example 2 Preparation of Compound Represented by Formula (I) (Step 2)

41.4 ml of 0.1M samarium iodide tetrahydrofuran solution (SmI ₂ , 0.1M solution in tetrahydrofuran) was stirred at 0 ° C. with 14-phenyltetradecan-2-one of formula IV. A solution of mg (1.1 mmol), 144.17 mg (1.44 mmol) of methyl methacrylate, and 105 μl (1.12 mmol) of t-butanol in 4 ml of anhydrous tetrahydrofuran were slowly added, followed by stirring at the same temperature for 5 hours. After completion of the reaction, the reaction solution was acidified with 1N HCl, the organic layer was extracted with diethyl ether, and washed with brine. The organic layer was dried over magnesium sulfate and concentrated, and the concentrated residue was separated by silica gel column chromatography (hexane: ethyl acetate = 3: 1) to obtain two isomers of compound (I) ( 2S ^* , 4S ^* )-I. 138 mg and ( 2R ^* , 4S ^* ) -I 185 mg were obtained (yield: 82%), respectively. The yield was also the same when using ( 2R, 3S ) -N-benzyl-ephedridyl methacrylate and ( 2S, 3R ) -Nt-butoxycarbonyl-noepedinyl methacrylate in addition to methyl methacrylate. .

The enantiomers of the two isomers were separated by high performance liquid chromatography using chiral column. In the case of ( 2S ^* , 4S ^* ) -I compounds, the isomers were separated at a ratio of 80:20 at 5.7 and 10.3 minutes retention time, and 30 (at 6.0 and 8.0 minutes) for ( 2R ^* , 4S ^* ) -I compounds. It was separated at a ratio of: 70.

( 2S ^* , 4S ^* )-I: ¹ H NMR (500 MHz, CDCl ₃ ): δ 7.15-7.29 (m, 5H), 2.76 (m, 1H), 2.59 (t, 2H, J = 7.7 Hz), 2.35 (dd, 1H, J = 9.3, 12.8 Hz), 1.62 (m, 1H), 1.59 (m, 4H), 1.40 (s, 3H), 1.27 (d, 3H, J = 7.2 Hz), 1.25-1.40 (m, 18H) ¹³ C NMR (125 MHz, CDCl 3): δ 179.7, 143.2, 128.6, 128.4, 125.7, 84.6, 42.0, 40.5, 36.2, 35.7, 31.7, 30.1, 29.8, 29.7, 29.6, 29.5, 27.2, 24.3, 16.3

( 2R ^* , 4S ^* )-I: ¹ H NMR (500 MHz, CDCl ₃ ): δ 7.15-7.29 (m, 5H), 2.80 (m, 1H), 2.59 (t, 2H, J = 7.7 Hz), 2.19 (dd, 1H, J = 9.1, 12.5 Hz), 1.68 (m, 1H), 1.64 (m, 4H), 1.35 (s, 3H), 1.26 (d, 3H, J = 7.6 Hz), 1.25-1.40 (m, 18H) ¹³ C NMR (125 MHz, CDCl ₃ ): δ 179.5, 143.2, 128.6, 128.4, 125.7, 84.4, 42.0, 41.9, 36.2, 35.2, 31.7, 30.0, 29.8, 29.7, 29.6, 29.5, 24.9 , 23.9, 15

The isomers of ( 2S ^* , 4S ^* ) -I are (I-1) and (I-2), and the ( 2R ^* , 4S ^* ) -I I-3) and (I-4).

Test Example 1 Activity and Toxicity Test

Transfection assay was used to confirm the activity of PPARδ and selectivity for PPARα and PPARγ for the four isomers of the compound of formula I (I-1, I-2, I-3, I-4). Confirmed by. In addition, an MTT assay was performed to confirm the toxicity of the cells at the concentration of the compound represented by the formula (I).

[Transaction Assay]

CV-1 cells were used as animal cell lines. Cells were cultured in DMEM medium in a 37 ° C. cell incubator containing 5% carbon dioxide. The medium contained 10% fetal bovine serum (FBS), 100 U / ml penicillin and 100 μg / ml streptomycin. On the first day of the experiment, CV-1 cells were seeded at 6,000 cells / well in 96 well plates. Plasmids expressing PPARs, plasmids expressing luciferase gene, and plasmids expressing beta-galactosidase when the PPARs bound to ligand were inoculated to cells inoculated on day 2 were transfected with Superfect (QIAGEN). Transfection). After 24 hours, the compounds of formula I dissolved in dimethylsulfoxide (DMSO) were diluted using medium and then treated with transfected cells at various concentrations. As a negative control, dimethyl sulfoxide was treated to a final concentration of 1%, and as a positive control, GW501516 compound was treated to a final concentration of 10 nM. After culturing for 24 hours, the cells were lysed using a lysis buffer, and luciferase activity was measured by adding luciferin in a luminometer. Beta-galactosidase activity was measured in an ELISA reader after addition of ONPG reagent. The luciferase level measured was corrected by the beta-galactosidase activity level, and the graph was used to calculate the EC ₅₀ value.

TABLE 1

As can be seen in Table 1, the isomer of (I-1) among the gammalactone compounds represented by the formula (I) according to the present invention has activity on PPARδ, EC ₅₀ value was 8.6μM . In addition, the compound showed no activity against PPARα and PPARγ, confirming the selectivity to PPARδ.

[Empty assay]

CV-1 cells were used as animal cell lines. Cells were cultured in DMEM medium in a 37 ° C. cell incubator containing 5% carbon dioxide. The medium contained 10% fetal bovine serum (FBS), 100 U / ml penicillin and 100 mg / ml streptomycin. On the first day of the experiment, CV-1 cells were seeded at 6,000 cells / well in 96 well plates. On the second day, the compounds of formula (I) dissolved in dimethyl sulfoxide (DMSO) were diluted using a medium, and then inoculated cells were treated at various concentrations. As a negative control, dimethyl sulfoxide was treated to a final concentration of 1%. After incubation for 24 hours, the empty reagent was added to the cells at a final concentration of 1 mg / ml. After 1 hour, the resulting purple crystals were dissolved in dimethyl sulfoxide and then absorbance at 570 nm was measured using an ELISA reader. The absorbance measured indicates the number of living cells, thereby confirming the cytotoxicity of Compound I compounds.

As a result, all of the prepared compounds showed no cytotoxicity even at 100 μM corresponding to 10 times or more of 8.6 μM of the EC ₅₀ value for the PPARδ activity of the compound of Formula I-1.

As described above, the pharmaceutical, functional cosmetics, functional foods and animal feed compositions of the present invention contains a gamma lactone compound or a pharmaceutically acceptable salt of the compound as an active ingredient peroxysomal proliferator activating receptor δ activating ligand It can be used to prevent and treat obesity, diabetes, hyperlipidemia, hypercholesterolemia, hyperlipoproteinosis, arteriosclerosis, hypertension, circulatory disease, ischemic heart disease or to strengthen muscles.

Claims (9)

Obesity, diabetes mellitus, hyperlipidemia, hypercholesterolemia, cyclopoproteinosis, arteriosclerosis, hypertension using the gamma lactone compound represented by the following formula (I) or a pharmaceutically acceptable salt of the compound as an active ingredient , Pharmaceutical composition for preventing and treating ischemic heart disease. [Formula I]

The method of claim 1, Gamma lactone (γ-lactone) compound represented by the formula (I) is characterized by having the following structure: obesity, diabetes, hyperlipidemia, hypercholesterolemia, cyclopoproteinosis, arteriosclerosis, hypertension, ischemic heart disease prevention and treatment Pharmaceutical composition for

delete Obesity, diabetes mellitus, hyperlipidemia, hypercholesterolemia, cyclopoproteinosis, arteriosclerosis, hypertension using the gamma lactone compound represented by the following formula (I) or a pharmaceutically acceptable salt of the compound as an active ingredient , Functional food for preventing ischemic heart disease. [Formula I]

Functional cosmetics for the prevention of obesity and the improvement of obesity as a gamma lactone (γ-lactone) compound represented by the formula (I) or a pharmaceutically acceptable salt of the compound as an active ingredient. [Formula I]

An animal feed composition comprising a gamma lactone compound represented by the following formula (I) or a pharmaceutically acceptable salt of the compound as an active ingredient. [Formula I]

delete a) reacting a compound of formula (IV) with a compound of formula (V) in the presence of a samarium iodide (SmI ₂ ) catalyst to prepare a lactone compound of formula (I); Method for producing a gamma lactone compound of formula (I) characterized in that it comprises a. [Formula I]

[Formula IV]

[Formula V]

[In the above formula, R is an alkyl group, aryl group, alkaryl group having 1 to 20 carbon atoms, the alkyl group, aryl group and alkylaryl group may further include a nitrogen atom.] An anabolic agent composition comprising a gamma lactone compound represented by the following formula (I) or a pharmaceutically acceptable salt of the compound as an active ingredient. [Formula I]