WO2012050370A2 - Composition for preventing or treating rheumatoid arthritis containing a gamma secretase inhibitor as an active ingredient - Google Patents

Abstract

The present invention relates to a composition for preventing or treating rheumatoid arthritis containing a gamma secretase inhibitor as an active ingredient. The gamma secretase inhibitor according to the present invention reduces the extent of arthritis induction and reduces the inflow of inflammatory cells into the knee joint, in a dose-dependent fashion, and also reduces the amount of cytokines and chemokines, and reduces the amount of expression of NICD (notch intracellular domain). Consequently, the gamma secretase inhibitor according to the present invention can be used as a medicinal product and a functional health food which is useful in the prevention or treatment of rheumatoid arthritis, since it regulates the activation, differentiation and growth of immune cells and hence has an outstanding effect in suppressing the production of NICD which plays a highly important role in notch signal transmission whereby inflammatory reactions occur.

Description

A composition for the prevention or treatment of rheumatoid arthritis containing a gamma-secretase inhibitor as an active ingredient

The present invention relates to a composition for preventing or treating rheumatoid arthritis, which contains a gamma-secretase inhibitor as an active ingredient.

Rheumatoid arthritis is an autoimmune disease, a chronic inflammatory disease associated with chronic inflammation of the joints. Often, inflammation spreads to tissues and other organs around the joints. In general, rheumatoid arthritis is a progressive disease that can lead to joint destruction and dysfunction, and joint inflammation associated with rheumatoid arthritis causes swelling, pain, stiffness and redness of the joint. Joint inflammation associated with rheumatoid arthritis can also occur in the tissues around the joint (tendons, ligaments, muscles). In some patients with rheumatoid arthritis, chronic inflammation destroys cartilage, bones and ligaments, causing joint deformation. Damage to joints can occur early in the disease and can be progressive. Progressive damage of the joints does not necessarily correlate with the degree of pain, stiffness or swelling in the joint.

On the other hand, gamma-secretase is a protein mainly studied in the treatment of Alzheimer's dementia and plays an important role in producing amyloid beta peptides, which are known to be the main cause of Alzheimer's dementia. Gamma-secretase is a co-complex proteolytic activator consisting of four proteins, for example presenilin, nicastrin, pen-2, aph-1. Presenilins represent a new group of anomalous aspartyl proteases that are essential for the activity of gamma-secretase and cleave in the trans membranes of these substrates to become polytopic membrane proteins themselves. Nicastin is a protein of relatively large molecular weight and is a structural unit of gamma-secretase present in the cell membrane. When present, the nicastrin acts as a receptor while the nicotine is directed out of the cell. It also forms bonds with some cells present in the cell membrane, including amyloid precursor protein (APP). Once the bond is formed, the amyloid precursor protein is moved to the active site of gamma-secretase, where the protein splits into two sites where the amyloid-beta exits the cell and the other site remains inside the cell. Amyloid-beta secreted out of the cells accumulates in the brain, creating plaques, which is a hallmark of Alzheimer's disease. The exact function of pen-2 and aph-1 is unknown (Wolfe MS, Biochemistry, 2006, 45, 7931-9).

Such gamma-secretase not only degrades amyloid beta precursor protein but also has various substrates such as Notch, CD44, N-cadherin, and IGF1-R. In particular, Notch has signaling mechanisms associated with inflammatory responses. Notch signaling regulates the differentiation, proliferation, survival and development of various cells. There are four notch receptors in mammalian cells ranging from Notch 1 to 4, and jagged-1,2 and delta-like-1,3. 4) It has a total of 5 ligands. When Notch receptors are activated after binding to ligands, two steps of proteolysis occur by the metalloproteases ADAM and gamma-secretase, which produce the notch intracelluar domain (NICD), which enters the nucleus and acts as a transcriptional regulator. do.

In particular, Notch signaling in mammals is involved in several stages in the development of T cells and B cells, and is also involved in the activation of T cells, the function of regulatory T cells and the differentiation of helper T cells. Notch signaling has also been reported to regulate differentiation and survival of dendritic cells (DCs) and to regulate the production of toll-like receptors (TLRs), which are abundantly produced in the joints of patients with rheumatoid arthritis (Amsen). , et al., Immunity, 2007, 27, 89-99, Amsen, et al., Cell, 2004, 117, 515-26, Eager, et al., Immunity, 2004, 20, 407-15, Fung, et al., Circulation, 2007, 115, 2948-56, Maillard, et al., Annu. Rev. Immunol., 2005, 23, 945-74, Osborne, et al., Nat. Rev. Immunol., 2007, 7 , 64-75, Ostrouhova, et al., J. Clin.Invest., 2006, 116, 996-1004, Skokos, et al., J. Exp. Med., 2007, 204, 1525-31, Tanigaki, et al., Nat. Immunol., 2007, 8, 451-6, Hu, et al., Immunity, 2008, 29, 691-703). Notch signaling is highly expressed in joint tissues of patients with rheumatoid arthritis and is known to increase the release of cytokines produced by rheumatoid arthritis (Nakazawa, et al., Arthritis and Rheumatism, 2001, 44, 1545-54). Ando, et al., Oncogene, 2003, 22, 7796-7803, Yabe, et al., J. Orthop. Sci., 2005, 10, 589-94).

There are more than 50 types of gamma-secretase inhibitors known to date, and recently, studies to use some of these gamma-secretase inhibitors as therapeutic agents for inflammation-related diseases have been steadily attempted. However, the anti-inflammatory effects of these substances, in particular the anti-inflammatory effects on joint inflammation associated with rheumatoid arthritis, are insignificant or unclear.

The present inventors are studying the anti-inflammatory effects of gamma-secretase inhibitors on joint inflammation associated with rheumatoid arthritis, and the gamma-secretase inhibitors reduce the degree of arthritis induction in a concentration-dependent manner, The present invention was confirmed to reduce influx, decrease the amount of cytokines and chemokines, and reduce the amount of expression of the notch intracelluar domain (NICD), thus completing the present invention.

Accordingly, the present invention is to provide a composition for the prevention or treatment of rheumatoid arthritis containing a gamma-secretase inhibitor as an active ingredient.

The present invention provides a composition for the prevention or treatment of rheumatoid arthritis containing a gamma-secretase inhibitor as an active ingredient.

The composition includes a pharmaceutical composition and a food composition.

Hereinafter, the present invention will be described in detail.

Gamma-secretase inhibitors according to the invention are DAPT, L-685,458, RO4929097, DBZ, Z-YIL-CHO, DAPM, compound E, CW, begacestat (GSI-953), semagacestat (LY450139), BMS 299897 and MRK One or more selected from the group consisting of 560, but is not limited thereto.

The DAPT (N-[(3,5-difluorophenyl) acetyl] -L-alanyl-2-phenyl] glycine-1,1-dimethylethyl ester) is a white solid, the molecular formula C ₂₃ H ₂₆ F ₂ N ₂ O ₄ , Molecular weight 432.46, CAS No. 208255-80-5.

L-685,458 ((5S)-(tert-butoxycarbonylamino) -6-phenyl- (4R) -hydroxy- (2R) -benzylhexanoyl) -L-leucy-L-phenylalaninamide) is represented by the molecular formula C ₃₉ H ₅₂ N ₄ O ₆ , Molecular weight 672.85, CAS No. 292632-98-5.

The RO4929097 is represented by the following Chemical Formula 1, a molecular formula C ₂₂ H ₂₀ F ₅ N ₃ O ₃ , molecular weight 469.4, CAS No. 847925-91-1.

[Formula 1]

DBZ (dibenzazepine; (S, S) -2- [2- (3,5-difluorophenyl) acetylamino] -N- (5-methyl-6-oxo-6,7-dihydro-5H-dibenzo [b, d ] azepin-7-yl) propionamide) is a white solid, formula C ₂₆ H ₂₃ F ₂ N ₃ O ₃ , molecular weight 463.4, CAS No. 209984-56-5.

The Z-YIL-CHO is represented by the following Chemical Formula 2, and is a white, lyophilized solid, having a molecular formula of C ₂₉ H ₃₉ N ₃ O ₆ , a molecular weight of 525.7.

[Formula 2]

The DAPM (N- [N-3,5-difluorophenacetyl] -L-alanyl-S-phenylglycine methyl ester) has the molecular formula C ₂₀ H ₂₀ F ₂ N ₂ O ₄ , molecular weight 390.4.

Compound E ((S, S) -2- [2- (3,5-difluorophenyl) -acetylamino] -N- (1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-benzo [e] [1,4] diazepin-3-yl) -propionamide) is an off-white powder, molecular formula C ₂₇ H ₂₄ F ₂ N ₄ O ₃ , having a molecular weight of 490.5.

The CW (3,5-bis (4-nitrophenoxy) benzoic acid) is a molecular formula C ₁₉ H ₁₂ N ₂ O ₈ , molecular weight 396.31, CAS No. 173550-33-9.

The begacestat (GSI-953) is represented by the following Chemical Formula 3, the molecular formula C ₉ H ₈ ClF ₆ NO ₃ S ₂ , molecular weight 391.74, CAS No. 769169-27-9.

[Formula 3]

The semagacestat (LY450139) is represented by the following formula 4, the molecular formula C ₁₉ H ₂₇ N ₃ O ₄ , molecular weight is 361.434.

[Formula 4]

The BMS 299897 is represented by the following Chemical Formula 5, the molecular formula C ₂₄ H ₂₁ ClF ₃ NO ₄ S, molecular weight 511.94, CAS No. 290315-45-6.

[Formula 5]

The MRK 560 (N- [cis-4-[(4-chlorophenyl) sulfonyl] -4- (2,5-difluorophenyl) cyclohexyl] -1,1,1-trifluoromethanesulfonamide) is represented by the following Chemical Formula 6, formula C ₁₉ H ₁₇ ClF ₅ NO ₄ S ₂ , molecular weight 517.92, CAS No. 677772-84-8.

[Formula 6]

Gamma-secretase inhibitors according to the present invention reduce the degree of arthritis induction, reduce the influx of inflammatory cells into the knee joint, reduce the amount of cytokines and chemokines, and express the notch intracelluar domain (NICD). Reduce the amount.

As described above, the gamma-secretase inhibitor according to the present invention has an excellent inhibitory effect on the production of NICD, which plays an important role in notch signal transduction, which triggers an inflammatory response by regulating the activation, differentiation and proliferation of immune cells. It can be used as a medicine and health functional food useful for the prevention or treatment of arthritis.

The composition of the present invention may contain one or more known active ingredients having a prophylactic or therapeutic effect of rheumatoid arthritis together with a gamma-secretase inhibitor.

The composition of the present invention may be prepared by including one or more pharmaceutically acceptable carriers in addition to the above-described active ingredients for administration. Pharmaceutically acceptable carriers may be used in combination with saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol and one or more of these components, if necessary, antioxidants, buffers And other conventional additives such as bacteriostatic agents can be added. Diluents, dispersants, surfactants, binders and lubricants may also be added in addition to formulate into injectable formulations, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. Furthermore, it may be preferably formulated according to each disease or component by a suitable method in the art or using a method disclosed in Remington's Pharmaceutical Science (Recent Edition), Mack Publishing Company, Easton PA.

The composition of the present invention can be administered orally or parenterally (eg, applied intravenously, subcutaneously, intraperitoneally or topically) according to the desired method, and the dosage is based on the weight, age, sex and health of the patient. The range varies depending on the diet, the time of administration, the method of administration, the rate of excretion and the severity of the disease. The daily dose of the gamma-secretase inhibitor is about 1 to 500 mg / kg, preferably about 150 to 250 mg / kg, preferably administered once to several times a day.

The composition of the present invention can be used alone or in combination with methods using surgery, hormonal therapy, drug therapy and biological response modifiers for the prevention or treatment of rheumatoid arthritis.

The composition of the present invention may be added to a dietary supplement for the purpose of preventing rheumatoid arthritis. When the gamma-secretase inhibitor of the present invention is used as a food additive, the gamma-secretase inhibitor can be added as it is or used with other foods or food ingredients, and can be appropriately used according to a conventional method. The mixed amount of the active ingredient may be appropriately determined depending on the purpose of use (prevention, health or therapeutic treatment). In general, the gamma-secretase inhibitors of the invention are added in amounts of up to 15% by weight, preferably up to 10% by weight, based on the raw materials in the manufacture of food or beverages. However, in the case of long-term intake for health and hygiene or for health control, the amount may be below the above range, and the active ingredient may be used in an amount above the above range because there is no problem in terms of safety.

There is no particular limitation on the kind of food. Examples of the food to which the substance can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, drinks, Alcoholic beverages and vitamin complexes, and includes all of the dietary supplements in the conventional sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates, etc. as additional components, as in the general beverage. Natural carbohydrates described above are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. As the sweetening agent, natural sweetening agents such as tautin and stevia extract, synthetic sweetening agents such as saccharin and aspartame, and the like can be used. The proportion of the natural carbohydrate is generally about 0.01 to 0.20 g, preferably about 0.04 to 0.10 g per 100 mL of the composition of the present invention.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, electrolytes, flavors, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, And a carbonation agent used for the carbonated beverage. In addition, the composition of the present invention may contain a pulp for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components can be used independently or in combination. The proportion of such additives is not critical but is usually selected in the range of 0.01 to 0.20 parts by weight per 100 parts by weight of the composition of the present invention.

The gamma-secretase inhibitor according to the present invention reduces the degree of arthritis induction, decreases the influx of inflammatory cells into the knee joint, reduces the amount of cytokines and chemokines, and activates, differentiates and proliferates immune cells. It is effective to reduce the amount of expression of the notch intracelluar domain (NICD), which plays an important role in notch signal transduction, which causes an inflammatory response.

1 is a diagram showing the degree of arthritis of the gamma-secretase inhibitor (DAPT) of the present invention.

Figure 2 is a diagram showing the quantification of the degree of arthritis of the gamma-secretase inhibitor (DAPT) of the present invention.

Figure 3 is a graph showing the degree of arthritis induction of the gamma-secretase inhibitor (DAPT) of the present invention in clinical scores.

Figure 4 is a microscopic view of histological inflammation in the knee joint of the rheumatoid arthritis animal model treated with gamma-secretase inhibitor (DAPT) of the present invention.

FIG. 5 shows cytokines (IL-6, IL-12, IL-17, TNF-a, INF-γ) in serum of a rheumatoid arthritis animal model treated with a gamma-secretase inhibitor (DAPT) of the present invention. This is a measure of the amount of chemokine (MCP-1) released.

Figure 6 is a diagram confirming the degree of NICD expression in the joints of the rheumatoid arthritis animal model treated with the gamma-secretase inhibitor (DAPT) of the present invention.

Figure 7 is a graph showing the degree of arthritis induction of the gamma-secretase inhibitors (L-685,458, RO4929097, DBZ) of the present invention.

FIG. 8 is a microscopic view of histological inflammation in the knee joint of a rheumatoid arthritis animal model treated with a gamma-secretase inhibitor of the present invention (L-685,458, RO4929097, DBZ).

9 shows cytokines (IL-6, IL-12, IL-17, TNF-a) in serum of rheumatoid arthritis animal models treated with gamma-secretase inhibitors (L-685,458, RO4929097, DBZ) of the present invention. , INF-γ).

Hereinafter, preferred examples are provided to aid in understanding the present invention. However, the following examples are merely provided to more easily understand the present invention, and the contents of the present invention are not limited by the examples.

Example 1 Effect of Gamma-Secretase Inhibitor (DAPT) on Arthritis Induction

In order to determine the effect of the gamma-secretase inhibitor (DAPT) of the present invention on the degree of arthritis induction, the following experiment was performed.

To determine whether arthritis induction is reduced by a gamma-secretase inhibitor in rheumatoid arthritis, a collagen induced arthritis model was constructed. In other words, an 8-week-old C57BL / 6J mouse was purchased and adjusted to the environment by supplying enough food and water for a month in a constant temperature / humidity animal breeding room that maintains a temperature of 22 ° C and a dark / light cycle of 12 hours. It was used for later experiments. Rheumatoid arthritis animal models were constructed of 12-week-old C57BL / 6J mice in the same manner as the nature protocol. Specifically, an emulsion made of the same amount of Bovine type II collagen (2 mg / mL, Chondrex, Japan) and Complete Freund's adjuvant (CFA, Chondrex, Japan) was injected into the tail and hips of C57BL / 6J mice by 0.1 mL. Primary immunization. After 3 weeks of first immunization, 0.1 mL of Bovine type II collagen (2 mg / mL, Chondrex, Japan) and an emulsion of Incomplete Freund's adjuvant (IFA, Chondrex, Japan) were injected into the tail and hip of the mouse for additional immunity. booster immunization). After additional immunization, the gamma-secretase inhibitor DAPT (Tocris bioscience, Cat. No: 2634, UK) was intraperitoneally injected into the mice for 4 weeks at 10 mg / kg and 20 mg / kg. After administration of DAPT for 4 weeks, arthritis induction was measured using the eXplore optix system by combining Cy5.5, a near-infrared fluorescence (NIRF) substance, with hydrophobically modified glycol chitosan (HGC), an angiogenic target substance. In addition, the degree of arthritis incidence was recorded by measuring the clinical score during the period of DAPT injection, and the clinical score was measured according to the example of the nature protocol.

The degree of arthritis induction and the result of quantification of DAPT, a gamma-secretase inhibitor of the present invention, are shown in FIGS. 1 and 2, respectively, and the result of measuring the degree of arthritis in clinical scores is shown in FIG. 3.

As shown in FIG. 1 and FIG. 2, DAPT, a gamma-secretase inhibitor, reduced the degree of arthritis induction in a concentration-dependent manner.

In addition, as shown in Figure 3, the degree of arthritis induction decreased depending on the concentration of DAPT over time.

Example 2 Effect of Gamma-Scretase Inhibitor (DAPT) on the Histological Inflammation of the Knee Joint

In order to determine the effect of the gamma-secretase inhibitor (DAPT) of the present invention on the degree of histological inflammation of the knee joint, the following experiment was performed.

The knee joint was extracted after intraperitoneally administering DAPT (10 mg / kg, 20 mg / kg) for 4 weeks to the animal model of rheumatoid arthritis prepared in Example 1. The extracted knee joint was fixed in 10% NBF (neutral buffered formalin). The fixed knee joint was decalcified using decalcifying solution lite (Sigma) to decalcify bone and make paraffin blocks. The prepared paraffin block joint tissue was made into 5 μm sections, stained with hematoxylin and eosin, and observed under a microscope.

The results are shown in FIG.

As shown in FIG. 4, influx of inflammatory cells occurred due to an inflammatory response of the knee joint in the knee joint tissue section of the rheumatoid arthritis animal model, but influx of inflammatory cells was decreased in the DAPT-treated group.

Example 3 Effect of Gamma-Secretase Inhibitor (DAPT) on the Release of Cytokines and Chemokines

In order to determine the effect of the gamma-secretase inhibitor (DAPT) of the present invention on the amount of release of cytokines and chemokines, the following experiment was performed.

Blood was collected after intraperitoneal administration of DAPT (10 mg / kg, 20 mg / kg) for 4 weeks to the animal model of rheumatoid arthritis prepared in Example 1. The collected blood was centrifuged at 13,000 rpm for 30 minutes to separate serum only. The isolated serum was cytokine (IL-6, IL-12, IL-17, TNF-a, INF-γ) and chemokine using Mouse Cytokine Panel 1-8 Plex (Millipore, MPXMCYTO-70K-08, USA) The amount of release of (MCP-1) was measured.

The results are shown in FIG.

As shown in Figure 5, the serum of the rheumatoid arthritis animal model was released a large amount of cytokines and chemokines, the amount of cytokines and chemokines was reduced in the DAPT group.

Example 4 Zero Effect of Gamma-Scretase Inhibitor (DAPT) on the Expression of NICD incense

In order to determine the effect of the gamma-secretase inhibitor (DAPT) of the present invention on the expression level of NICD, which plays an important role in Notch signal transduction, the following experiment was performed.

In the animal model of rheumatoid arthritis prepared in Example 1, joints were extracted after intraperitoneal administration of DAPT for 4 weeks. The extracted joints were crushed using a homogenizer (MagNA Lyser, Roche), and the tissue lysates (T-per, Thermo) were added and stirred in a 4 ° C refrigerator for 1 hour, followed by the addition of sample buffer solution at 100 ° C. Boiled for a minute. The prepared samples were confirmed by the western blot expression of NICD, which plays an important role in Notch signal transduction.

The results are shown in FIG.

As shown in FIG. 6, the expression level of Notch 1 was increased in the joints of the rheumatoid arthritis animal model, but the expression level of NICD was decreased in the DAPT-administered group.

Example 5 Effect of Gamma-Secretase Inhibitors (L-685,458, RO4929097, DBZ) on the Arthritis Induction Level

In order to determine the effect of the gamma-secretase inhibitors (L-685,458, RO4929097, DBZ) of the present invention on the degree of arthritis induction, DAPT (10mg / kg, 20mg / kg) instead of L-685,458 (Tocris bioscience, Cat.No: 2627, UK), RO4929097 (Selleck chemical, S1575 USA), DBZ (Calbiochem, Cat.No: 565789, Germary) (1 mg / kg) In the same manner as in Example 1, and confirmed the degree of arthritis.

The results are shown in FIG.

As shown in FIG. 7, the incidence of arthritis decreased in the order of the gamma-secretase inhibitor L-685,458> RO4929097> DBZ. In particular, the administration of L-685,458 (1mg / kg) of the present invention compared to the administration of DAPT (20mg / kg), the degree of arthritis was further reduced, RO4929097, DBZ also showed the same effect.

Example 6 Effect of Gamma-Secretase Inhibitors (L-685,458, RO4929097, DBZ) on the Histological Inflammation of the Knee Joint

In order to determine the effect of the gamma-secretase inhibitors (L-685,458, RO4929097, DBZ) of the present invention on the degree of histological inflammation of the knee joint, DAPT (10mg / kg, 20mg / kg) instead of L-685,458, RO4929097, DBZ (1mg / kg), except in the same manner as in Example 2, the histological inflammation of the knee joint in the animal model of rheumatoid arthritis microscopically Observed.

The results are shown in FIG.

As shown in FIG. 8, the influx of inflammatory cells occurred due to the inflammatory response of the knee joint in the knee joint tissue fragment of the rheumatoid arthritis animal model, but the influx of inflammatory cells in the group administered with L-685,458, RO4929097, and DBZ. This has been reduced.

Example 7 Effect of Gamma-Secretase Inhibitors (L-685,458, RO4929097, DBZ) on the Release Amount of Cytokines

Confirmation of the Gamma-secretase Inhibitor (L-685,458, RO4929097, DBZ) on the Release of Cytokines (IL-6, IL-12, IL-17, TNF-a, INF-γ) In Example 3, except that L-685,458, RO4929097, DBZ (1mg / kg) instead of DAPT (10mg / kg, 20mg / kg) as a gamma-secretase inhibitor in Example 3, In the same manner, the release amount of cytokines (IL-6, IL-12, IL-17, TNF-a, INF-γ) in the serum of the rheumatoid arthritis animal model was confirmed.

The results are shown in FIG.

As shown in Figure 9, the amount of cytokines (IL-6, IL-12, IL-17, TNF-a, INF-γ) was released in the serum of the rheumatoid arthritis animal model, L-685,458, RO4929097 , The amount of cytokines decreased in the group receiving DBZ.

Examples of preparations for the compositions of the present invention are illustrated below.

Formulation Example 1  : Preparation of Pharmaceutical Formulations

1. Preparation of powder

Gamma-secretase inhibitor 200 mg

Lactose 100mg

The above ingredients were mixed and filled in airtight cloth to prepare a powder.

2. Preparation of Tablets

Gamma-secretase inhibitor 200 mg

Corn Starch 100mg

Lactose 100mg

2 mg magnesium stearate

After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.

3. Preparation of Capsule

Gamma-secretase inhibitor 200 mg

Corn Starch 100mg

Lactose 100mg

2 mg magnesium stearate

After mixing the above components, the capsule was prepared by filling in gelatin capsules according to the conventional method for producing a capsule.

4. Preparation of Injectables

Gamma-secretase inhibitor 200 mg

Mannitol 100mg

Na ₂ HPO ₄ 12H ₂ O 2mg

Appropriate sterile distilled water for injection

Injectables were prepared by mixing the above ingredients per ampoule (2 mL) according to the conventional method for preparing injectables.

Formulation Example 2 Preparation of Food

Foods containing the gamma-secretase inhibitor of the present invention were prepared as follows.

1. Preparation of Cooking Seasonings

A health promotion cooking seasoning was prepared with a gamma-secretase inhibitor of 20-95 wt%.

2. Preparation of Tomato Ketchup and Sauce

Health promotion tomato ketchup or sauce was prepared by adding 0.2-1.0 wt% of gamma-secretase inhibitor to tomato ketchup or sauce.

3. Manufacturing of Flour Foods

Gamma-secretase inhibitor 0.5-5.0% by weight was added to the flour and using this mixture to prepare bread, cakes, cookies, crackers and noodles to produce health foods.

4. Preparation of soups and gravy

0.1-5.0% by weight of gamma-secretase inhibitor was added to the soup and gravy to prepare health promoting meat products, soups of noodles and gravy.

5. Preparation of Ground Beef

10% by weight of a gamma-secretase inhibitor was added to the ground beef to prepare a ground beef for health promotion.

6. Manufacture of Dairy Products

5-10% by weight of gamma-secretase inhibitor was added to the milk, and the milk was used to prepare various dairy products such as butter and ice cream.

Formulation Example 3 Preparation of Beverage

1. Preparation of Carbonated Drinks

10 to 15% of gamma-secretase inhibitor, 5 to 10% of sugar, 0.05 to 0.3% of citric acid, 0.005 to 0.02% of caramel, 0.1 to 1% of vitamin C, and 75 to 80% of purified water Mix to make syrup. The syrup was sterilized at 85 to 98 ° C. for 20 to 180 seconds, mixed with cooling water at a ratio of 1: 4, and 0.5 to 0.82% of carbon dioxide was injected to prepare a carbonated beverage containing a gamma-secretase inhibitor.

2. Manufacture of health drinks

Gamma-secretase inhibitor (2.5% solids, 97.16%), jujube extract (65 brix, 2.67%), overweight extract (70% solids, 0.12%), vitamin C (0.02%), calcium pantonthenate (0.02% ), Licorice extract (65% solids, 0.01% solids) was homogeneously blended and instant sterilized and then packaged in a small packaging container such as glass bottles and plastic bottles to prepare a healthy beverage.

3. Preparation of Vegetable Juice

0.5 g of gamma-secretase inhibitor was added to 1,000 mL of tomato or carrot juice to prepare vegetable juice for health promotion.

4. Preparation of Fruit Juice

0.1 g of gamma-secretase inhibitor was added to 1,000 mL of apple or grape juice to prepare fruit juice for health promotion.

Since the gamma-secretase inhibitor according to the present invention can effectively inhibit the production of NICD (Notch Intracelluar Domain), which plays an important role in the Nazi signal transduction that causes an inflammatory response by regulating the activation, differentiation and proliferation of immune cells. , A composition comprising a gamma-secretase inhibitor as an active ingredient can be usefully used for the treatment and prevention of representative rheumatoid arthritis of autoimmune diseases, and health functional food can be widely used for the prevention of rheumatoid arthritis. It is expected to be.

Claims (6)

1. A pharmaceutical composition for the prevention or treatment of rheumatoid arthritis, comprising a gamma-secretase inhibitor as an active ingredient.
2. The method of claim 1,

   The gamma-secretase inhibitor is DAPT (N-[(3,5-difluorophenyl) acetyl] -L-alanyl-2-phenyl] glycine-1,1-dimethylethyl ester), L-685,458 ((5S)-( tert-butoxycarbonylamino) -6-phenyl- (4R) -hydroxy- (2R) -benzylhexanoyl) -L-leucy-L-phenylalaninamide), RO4929097 of Formula 1, DBZ (dibenzazepine; (S, S) -2- [ 2- (3,5-Difluorophenyl) acetylamino] -N- (5-methyl-6-oxo-6,7-dihydro-5H-dibenzo [b, d] azepin-7-yl) propionamide), Z-YIL-CHO, DAPM (N- [N-3,5-difluorophenacetyl] -L-alanyl-S-phenylglycine methyl ester), compound E ((S, S) -2- [2- (3,5- difluorophenyl) -acetylamino] -N- (1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-benzo [e] [1,4] diazepin-3-yl) -propionamide), CW ( 3,5-bis (4-nitrophenoxy) benzoic acid), begacestat of formula 3 (GSI-953), semagacestat of formula 4 (LY450139), BMS 299897 of formula 5 and MRK 560 of formula 6 (N- [cis-4 At least one selected from the group consisting of-[(4-chlorophenyl) sulfonyl] -4- (2,5-difluorophenyl) cyclohexyl] -1,1,1-trifluoromethanesulfonamide) Characterized in that, the prevention or treatment of rheumatoid arthritis of the pharmaceutical composition.

   [Formula 1]

   

   [Formula 2]

   

   [Formula 3]

   

   [Formula 4]

   

   [Formula 5]

   

   [Formula 6]

   
3. The method of claim 1,

   A gamma-secretase inhibitor is characterized by inhibiting the production of the notch intracelluar domain (NICD), pharmaceutical composition for the prevention or treatment of rheumatoid arthritis.
4. A food composition for preventing rheumatoid arthritis, comprising a gamma-secretase inhibitor as an active ingredient.
5. The method of claim 4, wherein

   The gamma-secretase inhibitor is DAPT (N-[(3,5-difluorophenyl) acetyl] -L-alanyl-2-phenyl] glycine-1,1-dimethylethyl ester), L-685,458 ((5S)-( tert-butoxycarbonylamino) -6-phenyl- (4R) -hydroxy- (2R) -benzylhexanoyl) -L-leucy-L-phenylalaninamide), RO4929097 of Formula 1, DBZ (dibenzazepine; (S, S) -2- [ 2- (3,5-Difluorophenyl) acetylamino] -N- (5-methyl-6-oxo-6,7-dihydro-5H-dibenzo [b, d] azepin-7-yl) propionamide), Z-YIL-CHO, DAPM (N- [N-3,5-difluorophenacetyl] -L-alanyl-S-phenylglycine methyl ester), compound E ((S, S) -2- [2- (3,5- difluorophenyl) -acetylamino] -N- (1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-benzo [e] [1,4] diazepin-3-yl) -propionamide), CW ( 3,5-bis (4-nitrophenoxy) benzoic acid), begacestat of formula 3 (GSI-953), semagacestat of formula 4 (LY450139), BMS 299897 of formula 5 and MRK 560 (N- [cis-4) At least one member selected from the group consisting of-[(4-chlorophenyl) sulfonyl] -4- (2,5-difluorophenyl) cyclohexyl] -1,1,1-trifluoromethanesulfonamide) Characterized in that the food composition for the prevention of rheumatoid arthritis.

   [Formula 1]

   

   [Formula 2]

   

   [Formula 3]

   

   [Formula 4]

   

   [Formula 5]

   

   [Formula 6]

   
6. The method of claim 4, wherein

   A gamma-secretase inhibitor inhibits the production of NICD, characterized in that the food composition for preventing rheumatoid arthritis.

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