KR101533637B1 - Composition for prevention and treatment of diabetes mellitus - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for the prevention and treatment of diabetes comprising a compound represented by the formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient. Specifically, the compound represented by the formula (1) of the present invention has excellent activity of inhibiting PTPMT1 and can be usefully used as a composition for the prevention and treatment of diabetes.

Description

TECHNICAL FIELD The present invention relates to a composition for prevention and treatment of diabetes mellitus,

The present invention relates to a pharmaceutical composition for preventing and treating diabetes comprising a compound inhibiting PTPMT1 as an active ingredient.

Diabetes mellitus is divided into type 1 and type 2, type 1 diabetes is also called 'child diabetes' and is caused by failure to produce insulin at all. Type 2 diabetes, which is relatively insufficiently insulin-dependent, is characterized by insulin resistance (insulin resistance that lowers blood sugar and cells do not burn glucose effectively). Type 2 diabetes may be due to environmental factors such as high calorie, high fat, high protein diet, lack of exercise, and stress due to westernization of dietary habits. However, diabetes mellitus may also be caused by defects of specific genes, Infection, or drug.

Insulin resistance refers to a state in which the response to insulin under a given insulin concentration is lower than normal. After the development of radioimmunoassay for insulin in the 1960s, diabetic patients with increased blood insulin levels were found, and the insensitivity of insulin was reduced. Insulin resistance in the human body has recently been shown to be reduced in obese patients compared to normal people after insulin injections were directly measured in muscle glucose uptake. In addition, insulin resistance has been emphasized as a central cause of cardiovascular risk factors such as hypertension, hyperlipidemia, and atherosclerosis, as well as insulin resistance in peripheral tissues in type 2 diabetes. If the amount of insulin secreted by a normal person exceeds 60 U per day, the insulin resistance is considered to be insulin resistance. If the insulin requirement is 2 U / kg or more in diabetic patients, the insulin resistance is considered to be insulin resistance.

In the current diabetic drug therapy, re-evaluated metformin and thiazolidinedione (TZD) -related drugs have been shown to be useful in the treatment of type 2 diabetes mellitus, but the underlying cause of diabetes, including the development of insulin resistance, has not been addressed and many side effects have been reported There is a need to develop more efficacious and safe drugs that can drastically / fundamentally resolve insulin resistance. Some drugs partially overcome insulin resistance but have a weak effect.

Protein phosphorylation and dephosphorylation are widely known as important mechanisms used to induce signaling by cells among the various stages of cellular function. The signal from the cell is mainly transmitted through phosphorylation involved in kinase and dephosphorylation involving protein phosphatase. Particularly, the protein phosphatase that acts on the dephosphorylation process plays an important role in intracellular regulation and regulation of basic cell signaling mechanisms involved in metabolism, growth, proliferation and differentiation due to their unique activity in vivo have. Examples of the phosphatase include Cdc25 (Cdc25A, Cdc25B, Cdc25C) which removes phosphate from amino acid tyrosine, and protein phosphatases such as PTP1B, Prl-3, LAR, CD45, Yop, PP1 and VHR.

Hereinafter, conventional protein phosphatase activity will be described.

1. Cdc25

Cdc25 phosphatase belongs to the dual specific phosphatase because it acts on both the phosphorylated tyrosine or threonine of the protein substrate. The Cdc25 phosphatase is involved in CDK activity by removing inhibitory phosphate from the tyrosine and threonine residues of the cyclin dependent kinase (CDK) involved in the cell division cycle . Activation of CDK activates MPF (M promoter) in the cell cycle, resulting in increased mitotic activity of M, resulting in cell proliferation. Therefore, inhibiting the activity of the Cdc25 phosphatase inhibits cell division and thereby prevents cell proliferation. Cdc25 has been reported to have three types of Cdc25A, Cdc25B, and Cdc25C in human cells. Among them, Cdc25A or Cdc25B is highly expressed in cancer cells such as breast cancer, rectal cancer, non-Hodgkin's lymphoma, prostate cancer, pancreatic ductal adenocarcinoma, and lung cancer. Cdc25A is also known to be involved in the adhesion-dependent proliferation of myeloid leukemia cells, and is thought to be involved in the carcinogenesis process. Therefore, Cdc25 inhibitors can be a promising anticancer drug development target by inducing M phase arrest, and various studies have been conducted to investigate inhibitors thereof [Otani, T. et al., J. of Antibiotics 2000, 53 , 337; Lazo, JS et al., Bioorg . Med . Chem. Lett . 2000, 8 , 1451].

2. PTP1B

PTP1B is the intracellular protein phosphatase that has been purified and characterized for the first time among various protein phosphatases, has a molecular weight of ~ 50 Kda isolated from human placenta and has been cloned subsequently.

PTP1B is abundant in many human cells. In particular, PTP1B inhibits phosphorylation of the insulin receptor (IR) and phosphorylation of the insulin receptor substrate (IRS-1) in the signal transduction pathway of insulin. Kennedy and Ramachandran found that the sensitivity of insulin was increased by biochemical experiments using mice knocked out of the PTP1B gene ( Science 1999, 283 , 1544) And increased phosphorylation of insulin receptor protein in muscle cells. Type 2 diabetes is a non-insulin-dependent diabetes mellitus that normally secretes insulin in the pancreas that secretes insulin, but insulin resistance plays a role in insulin-resistant organs (muscle, liver, fat cells, etc.) It has been shown that the dephosphorylation of insulin receptor (IR) is directly related to insulin resistance and is involved in type 2 diabetes. Therefore, the PTP1B inhibitor acts on the dephosphorylation of insulin receptors, potentially overcoming insulin resistance and normalizing plasma glucose and insulin without inducing hypoglycemia. Therefore, as a treatment for type 2 diabetes, This is expected to be useful, and research on this field is very broad.

3. CD45

In order for the cells to grow normally, cells need to be balanced between very sophisticated signals. However, if this balance is broken and only the activation signal remains, the cells will grow uncontrollably causing disease. The CD45 is just stopping these signals. CD45 is a type of PTPase (Protein Tyrosine Phosphatase) located in the membrane and is known to be involved in the signal transduction of T cells or B cells. After removal of CD45 protein from mice, Janus kinase (JAK) and STAT (signal transducer and activators of transcription) were activated by cytokines and interferons, which inhibited the signal of cytokine by inhibiting Janus kinase .

4. LAR

The LTP of the PTPase family was thought to contribute to the physiological regulation of the insulin receptor in virgin cells. They reached the above conclusion by comparing the rate of dephosphorylation / deactivation of purified IR by LAR using recombinant PTP1B as well as cytoplasmic domains. Antisense inhibition has recently been used to study the effect of LAR on insulin signaling in rat liver tumor cell lines. Inhibition of LAR protein levels by about 60% and an increase of approximately 150% of autophosphorylation by insulin. However, only a moderate increase of 35% was observed in insulin receptor tyrosine kinase (IRTK) activity, while insulin-dependent phosphatidylinositol 3-kinase (PI 3-kinase) Was increased by 350%. The authors predicted that LAR could specifically dephosphorylate tyrosine residues that are crucial for PI 3-kinase activation on the insulin receptor itself or on subsequent substrates. Thus, it has been expected that LAR inhibitors may be usefully used as agents for the treatment of obesity, impaired glucose intolerance, diabetes mellitus, hypertension, or ischemic diesease of blood vessels. However, in spite of these many studies, it is still urgent to develop new materials because there are no substances that have been processed until the clinical process.

5. VHR

VHR, a bispecific phosphatase, activates extracellular signal receptor kinase 1 (ERK1), a kinase of the mitogen-activated protein kinase (MAPK) family, and ERK2 by mitogen signaling mitogenic signaling. Since VHR regulates the cell cycle, its inhibitors can be used for anticancer drugs as well as the Cdc25 inhibitors described above (Osada, H. et al., FEBS Letters 1995, 372 , 54).

6. Prl-3

So far, changes in gene levels in colon cancer have been mainly inactivation of tumor suppressor proteins, but they have been problematic in targeting new drug targets by small molecules. lately In a recent report (Saha et al., Science 2001, 294 , 1343), it has been found that a new phosphatase called Prl-3 is commonly overexpressed in various colon cancer cell metastases. Since the activity of Prl-3 appears to be essential for colon cancer metastasis, the development of an effective inhibitor of Prl-3 may be an important new drug target that can provide a new biomarker for the treatment of metastatic colon cancer.

7. PTPMT1

Dual-specificity protein tyrosine phosphatases (DUSPs) have unique enzymatic activities that promote phosphotyrosine and phosphotryronine dephosphorylation as well as phosphotyrosine residues in a variety of protein substrates . DUSPs are one of the major regulators of an abnormally regulated critical signaling pathway in a variety of diseases, including cancer, diabetes, and immune deficiency. Protein tyrosine phosphatase (PTPMT1) localized in mitochondria 1 belongs to DUSP and was discovered during screening of new DUSPs based on the sequence of catalytic motifs of PTEN (phosphatase and tensin homologues deleted on chromosome 10). PTPMT1 has also been identified in pancreatic islets and is known to be present in the inner membrane facing the mitochondrial matrix. Knockdown of PTPMT1 expression has substantially increased ATP levels in cells and insulin secretion in insulin alone producing cells (beta cells) in vivo, indicating that PTPMTl can be a potential target for the treatment of diabetes .

Recently, the X-ray crystal structure of mouse PTPMT1 has been reported in a state of resting and binding to phosphatidylinositol 5-phosphate, which acts as a substrate analogue. The three-dimensional structure of human PTPMT1 has also been reported in complexes with sulfate ion bonds in the active site as an effective surrogate for substrate phosphate groups. The presence of more structural information on the nature of PTPMT1 and small molecule ligand interactions facilitates the design of potential inhibitors that can be developed as anti-diabetic drugs. Nevertheless, the development of PTPMT1 inhibitors lags behind pharmaceutical and structural studies. The only known inhibitor of PTPMT1 to date is the dibiguanide group. Treatment of these inhibitors in the pancreatic islets of isolated rats resulted in increased insulin secretion in a dose-dependent manner, confirming that PTPMT1 should be a promising target in the development of anti-diabetic drugs. Despite their potency and selectivity in the inhibition of PTPMTl, it is difficult to apply these inhibitors to clinical practice due to the positivity of these molecular structures and the presence of long alkyl chains.

Thus, the present inventors have found that by using a structure-based drug design protocol using virtual screening and in vitro enzyme assay using docking simulations of PTPMT1, the binding free energy between PTPMT1 and the estimated ligand The inventors of the present invention have completed the present invention by identifying a new group of PTPMT1 inhibitors having an effect of high enzyme assay hit ratio by performing an accurate solvation model. The inhibitor of PTPMT1 according to the present invention can overcome the insulin resistance of the conventional diabetes therapeutic agent and thus can be effectively used as a therapeutic agent for diabetes.

It is an object of the present invention to provide a pharmaceutical composition for the prevention and treatment of diabetes comprising a phenyl derivative or a pharmaceutically acceptable salt thereof as an active ingredient.

Another object of the present invention is to provide a method for preparing 2- (3 - ([1,1'-biphenyl] -4-yl) -5,9-dimethyl- 6-yl) acetic acid (2- (3 - ([1,1'-biphenyl] -4-yl) -5,9-dimethyl-7-oxo-7H- yl) acetic acid, (E) -1- (4-butylphenyl) -5 - ((5- (2- Dihydro-pyrimidine-4,6 (1H, 5H) -dione ((E) -1- (4-butylphenyl) -5 - ) -2-thioxodihydropyrimidine-4,6 (1H, 5H) -dione), (E) -3- (5- (5-imino-7-oxo-3-phenyl-5H-thiazolo [ 2-yl) benzoic acid), (E) -4- (5 - ((8-methoxyquinolin-2-yl) methylene) (4-oxo-2-thioxothiazolidin-3-yl) benzoic acid ((E) 3-yl) benzoic acid, 4 - ((3- (4-ethoxyphenyl) -4-oxo-3,4,5,6,7,8- hexahydrobenzo [ Yl) thio) methyl) benzoic acid (4 - ((3- (4-ethoxyphenyl) -4-oxo-3,4,5,6,7,8- (3-oxobenzo [b] thiophen-2-yl) thio) methyl] benzoic acid), (E) -4- Yl) benzoic acid ((E) -4- (5 - ((3-oxobenzo [b] thiophen-2 (3H) -ylidene) methyl) furan-2-yl ) benzoic acid, (E) -2-hydroxy-4- (5 - ((8-methoxyquinolin- 3-yl) benzoic acid, (2E, 5E) - benzoic acid ((E) -2-hydroxy- (2E, 5E) -3-benzyl-2- (benzylimino) -5- (2-hydroxy-5-nitrobenzylidene) thiazolidin- -5- (2-hydroxy-5-nitrobenzylidene) thiazolidin-4-one, (3-chloro-4-methylphenyl) -3-methyl-lH-pyrazole-4 (5H) (5-oxo-1H-pyrazol-4 (5H) -ylidene) methyl) furan-2-yl) benzoic acid; 3- Yl) benzoic acid (3- (5 - ((E) - ((E) -4- 5-ylidene) methyl) furan-2-yl) benzoic acid), (Z) -2-hydroxy-5- (5- ((Z) -2-hydroxy-5- (5 - (((6-methoxy-1H- indol-3-yl) methylene) -4-oxo-2-thioxothiazolidin- 3-yl) benzoic acid and 5-benzyl-2- (phenoxymethyl) -3-phenylpyrazolo [l, 2-benzodiazepin- , 5-a] pyrimidin-7 (4H) -one, and 5-benzyl-2- (phenoxymethyl) -3-phenylpyrazolo [1,5- Or a pharmaceutically acceptable salt thereof as an active ingredient. The present invention also provides a pharmaceutical composition for the prevention and treatment of diabetes.

Another object of the present invention is to provide a health food for preventing and improving diabetes comprising the above-mentioned compound or a pharmaceutically acceptable salt thereof as an active ingredient.

In order to accomplish the above object, the present invention provides a pharmaceutical composition for preventing and treating diabetes comprising, as an active ingredient, a phenyl derivative represented by the following formula (1) for inhibiting PTPMT1 or a pharmaceutically acceptable salt thereof:

[Chemical Formula 1]

(Wherein R1, R2 and R3 are as described herein).

The present invention also provides a pharmaceutical composition for the prevention and treatment of diabetes comprising the compound represented by the following general formulas (2) to (13) or a pharmaceutically acceptable salt thereof as an active ingredient:

(2)

,

,

(3)

,

,

[Chemical Formula 4]

,

,

[Chemical Formula 5]

,

,

[Chemical Formula 6]

,

,

(7)

,

,

[Chemical Formula 8]

,

,

[Chemical Formula 9]

,

,

[Chemical formula 10]

,

,

(11)

,

,

[Chemical Formula 12]

; 및

; And

[Chemical Formula 13]

.

.

The present invention also provides a health food for preventing and improving diabetes mellitus containing the above compound or a pharmaceutically acceptable salt thereof as an active ingredient.

The compounds represented by the general formulas (2) to (13) of the present invention have a remarkable inhibitory effect on PTPMT1 in the PTPMT1 enzyme assay, effectively inhibiting PTPMT1 overexpressed in diabetes, and thus can be usefully used as a composition for the prevention and treatment of diabetes .

Figure 1 is a comparison of the PTPMTl inhibitor binding mode; (2) is black, (3) is gray, (4) is pink, (5) is orange, and (6) is purple.

Hereinafter, the present invention will be described in detail.

The present invention provides a pharmaceutical composition for the prevention and treatment of diabetes comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient:

[Chemical Formula 1]

(In the formula 1,

또는

이고;
R ₁ And when R < ₂ > is H, R < _{3 &}

or

ego;

,

또는

이며;When R ₁ is COOH and R ₂ is H, then R ₃ is

,

or

;

이고;When R ₁ is NO ₂ and R ₂ is OH, R ₃ is

ego;

,

또는

이며;When R ₁ is COOH and R ₃ is H, R ₂ is

,

or

;

이고,When R ₁ is -CH ₂ CH ₂ CH ₂ CH ₃ and R ₃ is H, R ₂ is

ego,

이며;When R ₂ is COOH and R ₃ is OH, R ₁ is

;

이다).When R ₂ is OH and R ₃ is COOH, then R ₁ is

to be).

The compound represented by the above-mentioned formula (1) can be obtained by reacting 2- (3 - ([1,1'-biphenyl] -4-yl) -5,9-dimethyl- 7-oxo-7H-furo [3,2-g] chromen-6-yl) acetic acid (2- -furo [3,2-g] chromen-6-yl) acetic acid)

(2)

.

.

The compound represented by the above formula (1) can be produced by reacting a compound represented by the following formula (3): (E) -1- (4-butylphenyl) -5 - (E) -1- (4-butylphenyl) -5 - ((5- (2-chloro-phenyl) -5-methylpyrrolidin- -5-nitrophenyl) furan-2-yl) methylene) -2-thioxodihydropyrimidine-4,6 (1H, 5H) -dione)

(3)

.

.

The compound represented by the above formula (1) is a compound represented by the following formula (4): (E) -3- (5 - ((5-Imino- (5-imino-7-oxo-3-phenyl-5H-pyrrolo [2,3-d] pyrimidin- thiazolo [3,2-a] pyrimidin-6 (7H) -ylidene) methyl) furan-2-yl) benzoic acid)

[Chemical Formula 4]

.

.

The compound represented by the above formula (1) is a compound represented by the following formula (5): (E) -4- (5 - ((8- 2-thioxothiazolidin-3-yl) benzoic acid) was used as the starting material. But is not limited to:

[Chemical Formula 5]

.

.

The compound represented by the above-mentioned formula (1) is preferably a 4 - (((3- (4-ethoxyphenyl) -4-oxo-3,4,5,6,7,8- (4 - ((3- (4-ethoxyphenyl) -4-oxo-3,4-dihydro- benzo [4,5] thieno [2,3- d] pyrimidin- , 5,6,7,8-hexahydrobenzo [4,5] thieno [2,3-d] pyrimidin-2-yl) thio) methyl) benzoic acid)

[Chemical Formula 6]

.

.

The compound represented by the above formula (1) is a compound represented by the following formula (7): (E) -4- (5- (3-oxobenzo [b] thiophene- (3-oxobenzo [b] thiophen-2 (3H) -ylidene) methyl) furan-2-yl) benzoic acid, But are not limited to:

(7)

.

.

(E) -2-hydroxy-4- (5 - ((8-methoxyquinolin-2-yl) methylene) -4- ((E) -2-hydroxy-4- (5 - ((8-methoxyquinolin-2-yl) methylene) -4-oxo-2-thioxothiazolidin- 3-yl) benzoic acid), but is not limited thereto:

[Chemical Formula 8]

.

.

(2E, 5E) -3-benzyl-2- (benzylimino) -5- (2-hydroxy-5-nitrobenzyl) (2E, 5E) -3-benzyl-2- (benzylimino) -5- (2-hydroxy-5-nitrobenzylidene) thiazolidin-4-one It does not:

[Chemical Formula 9]

.

.

The compound represented by the formula (1) is a compound represented by the following formula (10): (Z) -3- (5 - ((1- (3-Chloro-4-methylphenyl) (3-chloro-4-methylphenyl) -3-methyl-lH-pyrazole-4 (5H) 5-oxo-1H-pyrazol-4 (5H) -ylidene) methyl) furan-2-yl) benzoic acid)

[Chemical formula 10]

.

.

(E) - ((E) -4-oxo-2- (phenylimino) -3- (pyridine Yl) benzoic acid (3- (5 - ((E) - (E) -4-oxo-2- (phenylimino) - 3-ylmethyl) thiazolidin-5-ylidene) methyl) furan-2-yl) benzoic acid).

(11)

.

.

(Z) -2-hydroxy-5- (5 - ((6-methoxy-1 H-indol-3-yl) methylene) Methyl-4-oxo-2-thioxothiazolidin-3-yl) benzoic acid ((Z) -2-hydroxy-5- (5- -oxo-2-thioxothiazolidin-3-yl) benzoic acid), but is not limited thereto:

[Chemical Formula 12]

.

.

 The compound represented by the above-mentioned formula (1) can be obtained by reacting 5-benzyl-2- (phenoxymethyl) -3-phenylpyrazolo [1,5- a] pyrimidin- But are not limited to, 5-benzyl-2- (phenoxymethyl) -3-phenylpyrazolo [1,5-a] pyrimidin-7 (4H)

[Chemical Formula 13]

.

.

The compound according to the present invention may be prepared by a method known to those skilled in the art of organic synthesis, or may be commercially available, but is not limited thereto.

PTPMT1 was found in DUSP as a protein tyrosine phosphatase (PTPMT1) localized to mitochondrial 1 and screening for new DUSPs based on the catalytic motif sequence of PTEN (phosphatase and tensin homologues deleted on chromosome 10) . PTPMT1 has also been identified in pancreatic islets and is known to be present in the inner membrane facing the mitochondrial matrix. Knockdown of PTPMT1 expression substantially increased ATP levels in cells and insulin secretion in isolated insulin producing cells (beta cells) in vivo.

In addition, since PTPMT1 mRNA is expressed in the skeletal muscle, which is a major site of insulin resistance in type 2 diabetes, insulin resistance can be overcome by inhibiting PTPMT1. In patients with insulin resistance, reduced oxidative phosphorylation occurs, which means accumulation of lipids in muscle cells. Thus, inhibition of PTPMT1 increases production of ATP, resulting in increased substrate utilization, which not only reduces the accumulation of lipids in muscle cells, but also overcomes insulin resistance. Therefore, the compound of the present invention inhibiting PTPMT1 can overcome insulin resistance in type 2 diabetes.

Treatment of PTPMT1 inhibitors in the pancreatic islets of isolated rats resulted in an increase in insulin secretion in a dose dependent manner, and thus PTPMT1 inhibitory compounds could be usefully used as pharmaceutical compositions for the prevention and treatment of diabetes.

In one embodiment of the present invention, the compounds according to the present invention inhibited the catalytic activity of PTPMT1 over 70% in the PTPMT1 enzyme assay and significantly lowered the IC ₅₀ , indicating that PTPMT1 was significantly inhibited (See Table 1).

Also, in one embodiment of the present invention, such PTPMT1 inhibitory compounds are structurally diverse and have been found to have good physicochemical properties as drug substance.

Therefore, the compound according to the present invention significantly inhibits PTPMT1 and can increase insulin secretion, and thus can be effectively used for the prevention and treatment of diabetes. The diabetes is preferably type 2 diabetes.

The present invention includes not only the compounds represented by Chemical Formulas 1 to 13, but also pharmaceutically acceptable salts thereof, and possible solvates, hydrates, racemates, or stereoisomers thereof, which can be prepared therefrom.

The compound represented by the formula (1) of the present invention can be used in the form of a pharmaceutically acceptable salt, and as the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. Acid addition salts include those derived from inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid or phosphorous acid, and aliphatic mono- and dicarboxylates, phenyl-substituted alkanoates, hydroxyalkanoates, Dioleate, aromatic acid, aliphatic and aromatic sulfonic acids. Such pharmaceutically innocuous salts include, but are not limited to, sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate chloride, bromide, Butyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, succinate, maleic anhydride, maleic anhydride, , Sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, Methoxybenzoate, phthalate, terephthalate, benzene sulfonate, toluene sulfonate, chlorobenzene sulfide Sulfonate, methanesulfonate, propanesulfonate, naphthalene-1-sulphonate, naphthalene-1-sulphonate, , Naphthalene-2-sulfonate or mandelate.

The acid addition salt according to the present invention can be produced by a conventional method, for example, by dissolving the compound of formula (1) in an excess amount of an acid aqueous solution, and using this salt in a water-miscible organic solvent such as methanol, ethanol, acetone or acetonitrile Followed by precipitation.

By heating the same amount of the compound represented by the general formula (1) and the acid aqueous solution or alcohol, and then evaporating the mixture or drying the precipitated salt by suction filtration.

In addition, bases can be used to make pharmaceutically acceptable metal salts. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess amount of an alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the salt of the undissolved compound, and evaporating and drying the filtrate. At this time, it is preferable for the metal salt to produce sodium, potassium or calcium salt.

The composition containing the compound of the present invention preferably comprises 0.1 to 50% by weight of the composition based on the total weight of the composition, but is not limited thereto.

The pharmaceutical compositions of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of medicaments.

The pharmaceutical composition according to the present invention can be formulated in the form of oral, granule, tablet, capsule, suspension, emulsion, syrup, aerosol or the like oral preparation, external preparation, suppository and sterilized injection solution according to a conventional method Can be used. Examples of carriers, excipients and diluents that can be included in the composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

 In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may be formulated into the compositions of the present invention with at least one excipient such as starch, calcium carbonate, (sucrose), lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As a suppository base, witepsol, macrogol, tween 61, cacao paper, laurin, glycerol gelatin and the like can be used.

The composition of the present invention may be administered orally or parenterally, and any parenteral administration method may be used, and systemic administration or topical administration is possible, but systemic administration is more preferable, and intravenous administration is most preferable.

The preferred dosage of the composition of the present invention varies depending on the condition and the weight of the patient, the degree of disease, the type of drug, the route of administration and the period of time, but can be appropriately selected by those skilled in the art. However, for the desired effect, the composition of the present invention is preferably administered at 0.0001 to 0.03 g / kg per day, preferably 0.001 to 8 mg / kg per day. The administration may be carried out once a day or divided into several times. The dose is not intended to limit the scope of the invention in any way.

The present invention also provides a health food for preventing and improving diabetes comprising the compound represented by the formula (1) as an active ingredient.

The compound of the present invention represented by the above formula (1) significantly inhibits PTPMT1 in vitro, and thus can be effectively used as a health food for prevention and improvement of diabetes by increasing insulin secretion in the pancreatic beta-islets .

The compounds according to the present invention can be added to health supplements such as foods, beverages and the like for the purpose of preventing and improving diabetes.

There is no particular limitation on the kind of the food. Examples of the foods to which the above substances can be added include dairy products including dairy products, meat, sausage, bread, biscuits, rice cakes, chocolate, candies, snacks, confectionery, pizza, ramen and other noodles, gums, ice cream, Beverages, alcoholic beverages and vitamin complexes, dairy products, and dairy products, all of which include health functional foods in a conventional sense.

The compound represented by the formula (1) of the present invention can be added directly to food or used together with other food or food ingredients, and can be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (for prevention or improvement). Generally, the amount of the compound in the health food may be 0.1 to 90 parts by weight of the total food. However, in the case of long-term intake intended for health and hygiene purposes or for the purpose of controlling health, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.

The health functional beverage composition of the present invention is not particularly limited to the other ingredients other than the above-mentioned compounds as essential ingredients in the indicated ratios and may contain various flavors or natural carbohydrates as additional ingredients such as ordinary beverages. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavors (tau martin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be advantageously used as flavors other than those described above The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 of the composition of the present invention.

In addition to the above, the compound of the present invention can be used as a flavoring agent such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents and thickening agents (cheese, chocolate etc.), pectic acid and its salts, Salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated beverages and the like. In addition, the compounds of the present invention may contain natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks.

These components may be used independently or in combination. The ratio of such additives is not so critical, but is generally selected in the range of 0.1 to about 20 parts by weight per 100 parts by weight of the compound of the present invention.

Hereinafter, the present invention will be described in detail with reference to Examples, Experimental Examples and Preparation Examples.

The following Examples, Experimental Examples and Preparation Examples illustrate the present invention, but are not limited thereto.

< Example  1> PTPMT1 Virtual Search using Docking Simulation for

We have found that the most recent version of a chemical database distributed by Interbioscreen (htt: //www.ibscreen.com) containing approximately 477,000 synthetic and natural compounds is PTPMT1 containing 24 million compounds A docking library was built. Three-dimensional atomic coordinates in the X-ray crystal structure of human PTPMTl complexed with substrate ion-like sulfate ion were selected as the acceptor model in the docking simulated virtual search. After removing the solvent molecules, hydrogen atoms were added to each protein atom. We noted the proton addition of ionizable Asp, Glu, His and LYS residues in the original X-ray structure of PTPMT1. If one of the carboxylate oxygens in the side chain of the Asp and GIu residues is within 3.5 A distance, which is the limiting distance in the moderate intensity hydrogen bond that is generally accepted towards the hydrogen bond accepting functional group including the backbone aminocarbonyl oxygen The side chains of Asp and Glu residues were assumed to be neutral. Similarly, if the NZ atom is not close to a hydrogen-bond donating group, the lysine side chain is assumed to be proton-doped. The same procedure was also applied to determine the proton addition of ND and NE atoms in the His residue.

Prior to the virtual search in the docking simulation, we filtered on the basis of Lipinski's 'Rule of Five' to select only those compounds with physicochemical properties of potential drug candidates. Compounds containing reactive chemical groups such as aldehyde, acid halide and non-cyclic diastereomeric bonds have also been excluded because they can be positive in enzyme assays. All compounds contained in the docking library were then applied to the CORINA program to generate three-dimensional atomic coordinates, after which Gasteiger-Marsilli atomic charges were assigned. We have used the AutoDock program in the virtual screening of PTPMT1 inhibitors because it has been found in some target proteins that the performance of scoring functions for other PTPMT1 inhibitors is better. The AMBER force field parameter was used to calculate the van der Waals interaction and the internal energy of the ligand performed in the autodock program. Docking simulations using an auto-dock at the active site of PTPMT1 were performed to score and rank the compounds in the docking library according to the calculated binding affinity.

In a practical docking simulation of the compounds of the docking library, the present inventors utilized a novel empirical autodetection function formed by introducing a new solvent model for the compound. The modified score function is expressed by Equation (1) below.

[Equation 1]

Where W _vdw , W _hbond , W _elec , W _tor and W _sol are weighting factors of van der Waals, hydrogen bonds, electrostatic interactions, torsional term and desolvation energy of inhibitors, respectively . rij is associated with the depth of interaction represents the _{distance, A ij, B ij, C} ij and D _ij is the equilibrium separation and potential energy wells (potential energy well) between the two atoms. The hydrogen bonding term has an additional weighting factor, E (t), indicating an angle-dependent directionality. A cubic equation approach was applied to obtain the dielectric constant required to calculate electrostatic interactions between PTPMT1 and ligand molecules. In entropy terminology, N _tor represents the number of rotatable bonds in the ligand. In desolvation terms, _Si And V _i are the partial volume of the solvent parameter and atom i, respectively, and Occ _i ^max denotes the maximum atomic use. In calculating the molecular solvent free energy, we used the atomic parameters developed by Kang et al, because atoms other than carbon are not available in the latest version of the autodock. This variation of solvent-free energy terms increases the accuracy of the virtual search because underestimation of the ligand solvent often leads to overestimation of the ligand's affinity at many polar atoms.

As a result, of the 240,000 compounds that were subjected to the virtual search using the docking simulation, 200 highest score compounds were selected according to the best hit rate.

< Example  2> Enzyme assay of selected compounds through virtual search

Among the 240,000 compounds that have been subjected to the virtual search using the docking simulation, 200 highest score compounds were selected according to the best hit rate. 191 of these were obtained from compound suppliers and tested for PTPMT1 at a concentration of 50 μM.

The catalytic domain of human PTPMTl (PTPMTl-CD, residues 31-190) was subcloned into pET28a and overexpressed using E. coli BL21 (DE3) strain. Cells were grown for 16 hours at &lt; RTI ID = 0.0 &gt; 291K &lt; / RTI &gt; with 0.1 mM IPTG. His tagged PTPMT1-CD was purified by nickel-affinity chromatography and dialyzed against a buffer containing 20 mM Tris-HCL (pH 7.0), 0.2 M NaCl, 5% glycerol and 2 mM DTT. The in vitro inhibitory activity of 191 compounds selected through the above virtual search on recombinant human PTPMT1 was evaluated. During enzyme assay arbitrary inhibition by small molecule aggregates may occur. Therefore, among the 191 compounds selected through the above virtual search, it was further screened through spectrophotometric analysis that no aggregates were formed in the 5% DMSO solution. Six compounds were excluded from the enzyme inhibition assay because these compounds formed aggregates in 5% DMSO solution. Since the remaining 185 compounds were found to be highly water soluble, the effect of small molecule aggregates on the catalytic activity of PTPMT1 in the enzyme inhibition assay appears to be insignificant.

As a result, the present inventors identified twelve compounds represented by the following formula (2) or (13) inhibiting the catalytic activity of PTPMT1 at 70% or more at a concentration of 50 μM.

(2)

;

;

(3)

;

;

[Chemical Formula 4]

;

;

[Chemical Formula 5]

;

;

[Chemical Formula 6]

;

;

(7)

;

;

[Chemical Formula 8]

;

;

[Chemical Formula 9]

;

;

[Chemical formula 10]

;

;

(11)

;

;

[Chemical Formula 12]

; 및

; And

[Chemical Formula 13]

.

.

< Experimental Example  1 > IC ₅₀ Value analysis

The present inventors have found that Doughty-Shenton et al. Enzyme assays were performed by observing the degree of hydrolysis of OMFP (3-0-methylfluorescein phosphate) using the fluorescence spectrophotometer proposed by the present inventor. The purified PTPMT1-CD (100 nM), OMFP (25 μM) and inhibitory candidate material were incubated for 15 min in a reactive mixture containing 50 mM Tris-HCl (pH 7.0), 0.1 M NaCl, 5% glycerol and 5 mM DTT Respectively. This enzymatic reaction was stopped with the addition of sodium hydroxide (0.2M). Thereafter, the activity of the phosphatase was confirmed by measuring the absorbance which was changed by the hydrolysis of the substrate at 535 nm. The IC50 values of the inhibitors were determined from a direct regression curve analysis.

As a result, the present inventors determined IC ₅₀ values of the above 12 compounds. The inhibitory activity of the newly identified inhibitors is shown in Table 1. The terminal carboxylate moiety appears to serve as a surrogate for the substrate phosphate having a negative charge. To date, compound 2 or 13 is not known as a phosphatase inhibitor. As shown in Table 1, the twelve inhibitors had a potent effect on PTPMT1 with IC ₅₀ values ranging from 0.7 to 17.3 μM. These inhibitors are structurally diverse and have been shown to have good physico-chemical properties as drug candidates and can therefore be used as anti-diabetic drugs.

(2) to PTPMT1 For IC ₅₀  value compound IC ₅₀ (2) 2.9 ± 0.1 (3) 7.1 ± 0.3 Formula 4 7.3 ± 0.5 Formula 5 9.5 ± 1.2 6 10.7 ± 0.8 Formula 7 11.8 ± 1.4 8 12.4 ± 0.4 Formula 9 12.7 ± 1.4 10 14.2 ± 1.1 Formula 11 15.7 ± 0.9 Formula 12 16.5 ± 1.4 Formula 13 17.3 ± 0.7

< Experimental Example  2> PTPMT1  Inhibition of inhibitors On the mechanism  Structural Information

In order to obtain structural information on the inhibitory mechanism of the identified PTPMT1 inhibitors, the binding modes in the active site were investigated in a comparative fashion. Figure 1 shows the lowest energy structure of formulas 2-6 in the active site gap of PTPMT1 calculated with the modified autodock program. The results of this docking simulation are consistent in that the functional groups of similar chemical nature are released in a similar way as the relatively interactions with the protein groups. As shown in the overlapping of their docked structures, for example, the carboxylate group of the inhibitor, together with a hydrophobic group present between the two loop structures (the generic acid and the E-loop) on the active site, Point to the cysteine residue (Cys132). The proximity of inhibitor carboxylate groups to Cys132 in the structure of the best scoring indicates their need for inhibition of PTPMT1 as a surrogate for substrate phosphate group. To diagnose the possibility of allosteric inhibition of PTPMT1 by identified inhibitors, docking simulations were performed with coordinate maps for the receptor models, including all parts of the catalytic domain of PTPMT1. However, the binding sequence in which the inhibitor is located outside the active site was not observed in any new inhibitor. These results support that inhibitors impair the catalytic activity of PTPMT1 through specific binding to active sites.

The inventors have also identified detailed interactions involved in the stabilization of each inhibitor in the active site. Inhibitors appeared to be in close contact with Cys132-Arg138, Asp101-Met102 and Arg78-Phe79, which belong to PTP, the common acid and the E-loop, respectively. Carboxylate oxygen is also stabilized by hydrogen bonding with the framework amide group of Ser137. In the binding modes of formulas 1-12, the terminal carboxylate and aromatic group are stabilized by hydrogen bonding with amino acids and hydrophobic interactions around the PTP loop with the common acid and the nonpolar residue in the E-loop, respectively.

< Formulation example  1> Preparation of pharmaceutical preparations

1-1. Sanje  Produce

Compound 2 500 mg

Lactose 100 mg

10 mg of talc

The above components are mixed and filled in airtight bags to prepare powders.

1-2. Manufacture of tablets

Compound 2 500 mg

Corn starch 100 mg

Lactose 100 mg

2 mg of magnesium stearate

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

1-3. Manufacture of capsules

Compound 3 500 mg

Corn starch 100 mg

Lactose 100 mg

2 mg of magnesium stearate

The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

1-4. Injection preparation

Compound 4 500 mg

Sterile sterilized water for injection

pH adjuster

(2 ml) per ampoule in accordance with the usual injection method.

1-5. Liquid  Produce

Compound 5 100 mg

10 g per isomer

5 g mannitol

Purified water quantity

Each component was added to purified water in accordance with the usual liquid preparation method and dissolved, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, and purified water was added thereto. The whole was adjusted to 100 ml with purified water, The liquid is prepared by sterilization.

< Formulation example  2> Manufacture of health food

Compound 6 1000 mg

Vitamin mixture quantity

70 [mu] g of vitamin A acetate

Vitamin E 1.0 mg

0.13 mg of vitamin

0.15 mg of vitamin B2

0.5 mg vitamin B6

0.2 [mu] g vitamin B12

10 mg vitamin C

Biotin 10 μg

Nicotinic acid amide 1.7 mg

50 mg of folic acid

Calcium pantothenate 0.5 mg

Mineral mixture quantity

1.75 mg of ferrous sulfate

0.82 mg of zinc oxide

Magnesium carbonate 25.3 mg

15 mg of potassium phosphate monobasic

Secondary calcium phosphate 55 mg

Potassium citrate 90 mg

100 mg of calcium carbonate

24.8 mg of magnesium chloride

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.

< Formulation example  3> Manufacture of health drinks

Compound 6 1000 mg

Citric acid 1000 mg

100 g of oligosaccharide

Plum concentrate 2 g

Taurine 1 g

Purified water was added to a total of 900 ml

The above ingredients were mixed according to the usual health drink manufacturing method, and the mixture was stirred and heated at 85 for about 1 hour. The resulting solution was filtered to obtain a sterilized 2-liter container, which was sealed and sterilized, And used for manufacturing.

Although the composition ratio is relatively mixed with the ingredient suitable for the favorite drink, it is also possible to arbitrarily modify the blending ratio according to the regional or national preference such as the demand class, demand country, use purpose, and the like.

< Formulation example  4> Manufacture of other health food

4-1. Manufacturing of beverages

Honey 522 mg

5 mg &lt; RTI ID = 0.0 &gt;

Nicotinic acid amide 10 mg

3 mg of sodium riboflavin hydrochloride

Pyridoxine hydrochloride 2 mg

Inositol 30 mg

Orthoic acid 50 mg

Compound 2 0.48 to 1.28 mg

200 ml of water

A beverage was prepared using the above-mentioned composition and content by a conventional method.

4-2. Of chewing gum  Produce

Gum base 20%

Sugar 76.36 ~ 76.76%

Compound 3 0.24 to 0.64%

Fruit flavor 1%

Water 2%

Chewing gum was prepared using the above-mentioned composition and content by a conventional method.

4-3. Manufacture of candy

Sugar 50 to 60%

Syrup 39.26 ~ 49.66%

Compound 4 0.24 to 0.64%

Orange fragrance 0.1%

The composition and the content of the candy were prepared using a conventional method.

4-4. Manufacture of flour food products

0.5 to 5 parts by weight of Compound 7 were added to 100 parts by weight of wheat flour, and bread, cake, cookies, crackers and noodles were prepared using this mixture to prepare foods for health promotion.

4-5. dairy product( dairy products )

Compound 9, 5 to 10 parts by weight was added to 100 parts by weight of milk, and the milk was used to make various dairy products such as butter and ice cream.

4-6. Manufacture of wires

Brown rice, barley, glutinous rice, and yulmu were alphalized by a known method and dried, and the powder was made into a powder having a particle size of 60 mesh by a pulverizer. Black beans, black sesame seeds, and perilla seeds were steamed and dried by a known method, and then powdered with a particle size of 60 meshes by a grinder. The grains and seeds prepared above were mixed with the compound 1 of the present invention in the following proportions.

Brown rice 30%

Yulmu 15%

Barley 20%

Perilla 7%

Black soybeans 7%

Black sesame 7%

Compound 5 3%

Manure 0.5%

0.5%

Claims (7)

delete A pharmaceutical composition for the prevention and treatment of diabetes comprising a compound of the following formula (2) or a pharmaceutically acceptable salt thereof as an active ingredient:
(2)

.
The pharmaceutical composition according to claim 2, wherein the diabetes is type 2 diabetes.
delete A health food for the prevention and improvement of diabetes comprising a compound of the following formula (2) or a pharmaceutically acceptable salt thereof as an active ingredient:
(2)

.
The health food for diabetes prevention and improvement according to claim 5, wherein the diabetes is type 2 diabetes.
delete