KR101551187B1 - Indazole derivatives selectively inhibiting the activity of Janus kinase 1 - Google Patents

Abstract

INDUSTRIAL APPLICABILITY The present invention relates to an indazole derivative having an activity of selectively inhibiting the activity of Janus Kinase 1 and its use as a therapeutic agent for rheumatoid arthritis.

Description

Indazole derivatives selectively inhibit the activity of Janus kinase 1 (Janase kinase 1)

INDUSTRIAL APPLICABILITY The present invention relates to an indazole derivative and its selective inhibition of the activity of Janus kinase 1 and its use as a therapeutic agent for rheumatoid arthritis.

An immunosuppressant is a generic term for an agent that inhibits the immune response of a living body. It is mainly used as a drug for the treatment and prevention of organ transplant rejection and autoimmune diseases.

Organ transplantation is the best treatment for end-stage organ failure, unless there is development of new therapies, which is a bright field of view in terms of the need and demand. Immunosuppressants are used to protect implanted tissues / organs from immune rejection, and there are now 25 organ transplants.

An autoimmune disease is an autoimmune disease that occurs when an immune system of an individual recognizes an endogenous protein as an external antigen, or an immune-functioning T cell is destroyed by the action of an antigen to destroy the tissue. The disease is caused by rheumatoid arthritis, Psoriasis, Crohn's disease, and the like. It has not been clarified about the clear pathogenesis and mechanism. Currently, it is reported that the effect of the immunosuppressive drug such as steroids is temporarily improved without the fundamental treatment, and immunosuppressive therapy has been proved to be effective in suppressing such autoimmune response.

Immunosuppressants are largely divided into chemical immunosuppressants and biological immunosuppressants.

Chemical immunosuppressants are the most commonly prescribed immunosuppressants for the treatment of organ transplant rejection and autoimmune diseases, and Fujisawa's FK506, which has almost the same mechanism of action as Cyclosporine A (85% of the total) of Novartis is representative. However, the signal transduction pathway, which is the point of drug action of the chemical immunosuppressant, is not specific to activated immune cells, and thus has various problems such as nonselective signal transduction inhibition, which causes various serious side effects.

In contrast, biological immunosuppressants include TNF-α inhibitors (etanercept, infliximab, and adalimumab), which are used as therapeutic agents for autoimmune diseases such as rheumatoid arthritis (RA), psoriasis and Crohn's disease. Daclizumab (Zenapax) and Basiliximab (Simulect) are also available for treatment of monoclonal antibodies or polyclonal antibodies to organ systems in organ transplant recipients. ABXCBL and the like are being developed. In addition, antibodies to B cells and T cells are the most clinically used antibodies. In addition, IL-2 receptor (CD25) and CTLA -4, antibodies against CD40 and CD40 ligands, and the like. However, biological immunosuppressants have a number of problems, including: 1) TNF-α has no effect on all RA patients and has more than 30% non-compliance; 2) selective immunity to activated immune cells 3) It is very expensive for development and treatment. Immunosuppressive drugs should be taken over a lifetime, which can give a great economic burden to patients. 4) Because they are developed as injections Patient compliance is very low and there is a risk of injection side effects and infection.

As the number of transplantation surgeries increases and the patient's survival is prolonged, recent efforts to minimize the side effects of immunosuppressants and improve the 'quality of life' of patients undergoing transplant surgery are required. The ideal immunosuppressive agent should be a specific immunosuppression that selectively inhibits only the clone that specifically reacts with the antigen, but it is not yet a stage that can be used clinically, and the currently used agent has a specificity for the activated immune cells As a non-immunosuppressive agent, it inhibits the host's normal immune response and causes lethal side effects in the kidneys and liver, and causes complications such as infectious diseases and malignant tumors. Until now, a single drug can not achieve sufficient efficacy to prevent rejection, so it is necessary to combine several methods. Therefore, the demand for immunosuppressive drugs that organ transplant patients have to use for a lifetime is also increasing, but the demand for the development of new immunosuppressants is increasing due to the side effects and the high price of long-term use.

Immune cells are signaled by the cytokine, and therefore, there is a specific receptor for each cytokine. The cytokine receptors that bind to IL-2, IL-4, IL-7, IL-9, IL-15 and IL-21 have a central part called the common cytokine receptor γ-chain γc family, which are known to play a crucial role in the formation and function of lymphocytes. Therefore, mutations in the gene coding for γc may occur, or the cytoplasmic tyrosine kinase JAK3 (janus kinase 3) (SCID) is known to occur when a mutation in a gene coding for a mutation occurs. As a result, studies on the development of immunosuppressive agents using a substance capable of selectively inhibiting the activity of JAK3 have been actively attempted.

Many pharmaceutical companies have competitively developed JAK3 inhibitors in anticipation of the development of therapeutic agents for immune diseases such as rheumatoid arthritis (RA) by the selective inhibition activity of JAK3. As a result, recently, Pfizer's Xelianz (Tofacitinib , CP-690,550) has been approved by the FDA as a treatment for rheumatoid arthritis.

However, CP-690,550, which was known to have JAK3 selective inhibitory activity at the early stage of development, has since been found to be a pan-JAK inhibitor with overall inhibitory activity against the JAK kinase family ( Science 2003, 302: 875) Has been controversial as to whether the inhibitory activity of any JAK isozyme is the most important factor.

A recent series of studies ( Cell 1998, 93: 373; Immunity 2000, 13: 561; Cell 1998, 93: 385; Lancet 2008, 371: 987; Chem . Biol . 2011, 18: 314). The JAK kinase, which plays a key role in the signal transduction pathway by γc-related cytokine receptors, is JAK1 and JAK3 plays only a very minor role.

Therefore, JAK inhibitor development research in the field of RA therapy focuses on JAK1 rather than JAK3, and recently JAK1 selective activating substances have been reported (J. Med. Chem. 2012, 55: 5901; J. Med Chem., 2012, 55: 6176).

On the other hand, JAK2 plays a crucial role in the signal transduction process related to erythropoiesis. Thus, when JAK2 activity is inhibited, hematopoiesis such as anemia, thrombocytopenia, and leukopenia may occur. Inflammatory cytokines IL-1, TNF-α, and TGF-β are found in 8-70% of autoimmune disease patients and in 8-71% of organ transplant patients. Etc. can reduce red blood cell production in anemia situations. Therefore, in the development of immunosuppressants to be taken over a lifetime, selectivity to JAK2 must be ensured to prevent the patient's inflammatory anemia deterioration.

[Prior Patent Literature]

Korean Patent Publication No. 1020100032886

DISCLOSURE OF THE INVENTION The present invention has been made in view of the above problems, and it is an object of the present invention to develop a new therapeutic agent for rheumatoid arthritis by developing a novel inhibitor with enhanced JAK1 / JAK2 selectivity using an indazole derivative, It is aimed at.

In order to accomplish the above object, the present invention provides a compound of the following general formula (I) or a pharmaceutically acceptable salt thereof:

[Chemical Formula 1]

In the above formula, n is an integer of 1 or 2, and R 2 is hydrogen or an amino group.

The present invention also provides a composition for inhibiting Janus kinase 1 comprising the compound of the present invention as an active ingredient.

The present invention also provides a pharmaceutical composition comprising the compound of the present invention as an active ingredient.

In addition, in one embodiment of the present invention, the composition is preferably but not limited to have a therapeutic or preventive effect on rheumatoid arthritis.

The present invention also provides a food composition for improving rheumatoid arthritis comprising the compound of the present invention as an active ingredient.

The present invention also relates to a process for the preparation of 4- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2- A Suzuki coupling reaction and a Boc-protecting group elimination reaction of the compound of the present invention.

The compositions of the present invention may be administered topically, orally, or parenterally. For example, the composition can be administered extracorporeally, into the skull, into the vagina, into the rectum, into the skin, into the heart, into the heart, into the stomach, intravenously, intramuscularly, intraperitoneally, percutaneously, intranasally, ≪ / RTI > As used herein, "intracranial administration" refers to administration of a substance directly to the brain via a catheter or needle, such as through an intrathecal, intracisternal, intracisternal, or transsphenoidal ).

The parenteral administration of the composition, when used, is generally characterized by injection. The injectable can be prepared in conventional form, as a liquid solution or suspension, in the form of a solid suitable for suspension in liquid before injection, or as an emulsion. More recently, the approach for improved parenteral administration involves living to maintain a constant dose by using a slow release or sustained release system (see U.S. Patent No. 3,610,795).

The exact amount of composition required will depend on the patient, age, weight and general condition of the patient, the severity of the allergic disorder being treated, the manner of administration, etc., depending on the patient. It is therefore impossible to determine the exact amount for all compositions. However, the appropriate amount can also be determined by those skilled in the art using only basic experimentation according to the teachings provided in the art.

The preferred dosage of the compound 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 compound of the present invention is preferably administered at 0.0001 to 100 mg / kg, preferably 0.001 to 100 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 pharmaceutical compositions of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of pharmaceutical compositions.

The pharmaceutical dosage forms of the compositions of the present invention may also be used in the form of their pharmaceutically acceptable salts and may be used alone or in combination with other therapeutically active compounds as well as in a suitable set.

And may be formulated in the form of oral preparations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups and aerosols, external preparations, suppositories and sterilized injection solutions. Examples of carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, 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 contain 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 administration 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. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The present invention provides a health supplement comprising a compound of the present invention and a pharmaceutically acceptable food-aid additive exhibiting the effect of improving rheumatoid arthritis. Examples of foods to which the compound of the present invention can be added include various foods, beverages, gums, tea, vitamin complexes, health supplements and the like, and they can be used in powder, granule, tablet, have.

It can also be added to foods or beverages for the purpose of improving rheumatoid arthritis. At this time, the amount of the compound in the food or drink may be 0.01 to 15% by weight of the total food, and the health beverage composition may be added in a proportion of 0.02 to 5 g, preferably 0.3 to 1 g, have.

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 thickening agents, 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 flesh for the production of natural fruit juices and fruit juice drinks and vegetable drinks.

These components may be used independently or in combination. The proportion of such additives is not so critical, but is generally selected in the range of 0 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.

In the present invention, an inhibitor which is more selective than JAK2 against JAK1 was developed. For this purpose, it was first necessary to understand the specificity of the JAK1 substrate binding site compared to the substrate binding sites of other isozymes. By comparing the three-dimensional structure of each isozyme binding to the same substrate or inhibitor, .

Therefore, the three-dimensional structure of JAK1 (PDB ID = 3EYH), JAK2 (PDB ID = 2B7A), JAK3 (PDB ID = 3LXL) and Tyk2 (PDB ID = 3LXP) combined with C-2 methyl imidazopyrrolopyridines (CMP) After downloading, this structure was superimposed together using the 'protein structure alignment' tool of the molecular modeling software Maestro [Fig. 1]. The structure of JAK 1 ~ 3, except for the structure of tyk2, which is relatively less structurally similar to the rest of the kinases, can be seen to be superimposed very well, but their structure is finely, but very characteristic, in the so-called glycine- And it was observed that there was a significant difference. That is, JAK1 has a relatively small substrate binding site as compared to JAK2 and JAK3, resulting in hydrogen bonding of Asp1003 and His885 at this site. In addition, Phe860, an amino acid residue of JAK2 corresponding to His885 of JAK1, can be seen to place an aromatic chain away from Asp976 in the N-terminal lobe. As a result, the substrate binding site of JAK2 has a relatively open structure (Fig. 1).

Due to this structural feature, JAK1 has a small cavity with a size suitable for binding with a short straight-chain functional group directly below the substrate binding site (Fig. 2). Asp1003 and His885, which form hydrogen bonds, are located in the lower part of the cavity, it is expected to maximize the bonding force with the cavity by introducing the polar functional group at the end of the straight type functional group bonding to the cavity.

Based on these findings, the inventors of the present invention have devised a novel JAK1 selective inhibitor that incorporates propargyl alcohol and but-3-yn-1-ol as short chain linear functional groups capable of effectively binding to the cavity of JAK1, 3-yn-1-ol is introduced at the 3-position of indazole in the state of the scaffold which hydrogen bonds the indazole-7-carboxamide nucleotide with the kinase hinge motif as shown in Formula 1, In order to induce the hydrophobic bond with the substrate binding site at position 5, another indazole ring was bonded to the JAK substrate binding site.

[Chemical Formula 1]

Thus, in the present invention, an indazole derivative represented by Chemical Formula 1 is chemically synthesized to verify the inhibition of JAK1, JAK2, JAK3, and Tyk2 [Fig. 3] to [Fig. 5] Discovered new uses and completed inventions.

The synthesis of the indazole derivative was applied in accordance with a known synthesis method (Fig. 3 to Fig. 5). Gritty above, 5- (4'-indazolyl) indazole -7-carboxamide derivative (1a ~ 1f) is 3-alkynolyl-5-bromo- indazole-7-carboxamides (2a ~ 2c) and 4- (4,4,5 Suzuki coupling reaction and Boc-protecting group removal of 5-tetramethyl-1,3,2-dioxaborolan-2-yl) -1H-indazole derivatives ( 3a to 3b ) [Fig. 3].

At this time, the main intermediates 3-alkynolyl-5-bromo-indazole-7-carboxamides ( 2a to 2c ) and 4- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan- -1H-indazole derivatives ( 3a to 3b ) were synthesized starting from methyl 2-amino-3-methylbenzoate 4 [FIG. 4] and 2,4-dinitrotoluene 8 [FIG.

First, Compound 4 was reacted with NBS to selectively carry out a bromination reaction at the para position of the amino group to obtain Compound 5 in a yield of 62% (FIG. 4). Synthesis of Indazole rings was accomplished by acetalization followed by diazotization, as known (WO 2009146406, 2009), resulting in a mixture of 6a (46% yield) and 6b (35% yield). The acetyl group of compound 6a could be converted quantitatively to compound 6b by treatment with 6N HCl in a methanol solvent. Compound 6b thus obtained was converted to amide 2a at a yield of 95% by a series of hydrolysis, amidation and Boc-protection. Compound 2a was treated with NIS to give 3-iodoindazole 7 in 54% yield. The desired compounds indazolyl-3-alkynols 2b and 2c were obtained in a yield of 36% and 30% respectively by Sonogashira coupling reaction of compound 7 with propargyl alcohol or but-3-yn-1-ol. 4]

Meanwhile, 4- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) -1H-indazole derivatives ( 3a to 3b ) were synthesized using a synthesis method similar to the above ]. That is, the market to buy a 2,4-dinitrotoluene (8) of NBS and the reaction was then brominated, nitro 3-bromo-4-methyl- 5-nitroaniline by reduction of the iron powder and using 3-bromo-2 -methyl-5-nitroaniline in a 3: 1 ratio (74% yield). After the amino group obtained by the reduction reaction was Boc-protected and the remaining nitro group was reduced again, the desired compound 9a was obtained together with the positional isomer 9b and each was separated by silica gel column chromatography. By cyclization of the compound 9a under the reaction conditions, such as the above it was able to synthesize the indazole compounds 11a and 11b (81% yield), where compound 11a was converted to 11b quantitatively by a deacetylation reaction. Finally, the desired compound 4- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) -1H-indazole derivative 3a was prepared by reacting compound 11b with bis (pinacoloato) diboron and PdCl ₂ dppf) ₂ in the presence of KOAc was obtained in a yield of 36%. On the other hand, Compound 3b was synthesized starting from commercially available 3-bromo-2-methylaniline 10 as a starting material in a total yield of 44% by the synthesis method as described above.

The JAK kinase inhibitory activity of the synthesized indazole derivatives was measured using the Z'-LYTE ™ Kinase Assay Kit-Tyr 6 Peptide (JAK1 to JAK3) and Tyr 3 Peptide (Tyk2) (Invitrogen) , And% inhibition of each JAK isoenzyme by indazole derivatives at a concentration of 10 μM are summarized in FIG.

As expected, the synthesized indazole derivatives showed almost no inhibitory activity on JAK2, and in particular, JAK1 selectivity for JAK2 in Compound 1d was found to be 67 times or more. In addition, in the case of the derivatives 1a and 1b having no alkynol substituent at the position 3, the JAK1 inhibitory activity was not large, but the derivatives 1c to 1f having the alkynol substituent showed significant JAK1 inhibitory activity. Especially, propargyl alcohol was substituted The effect of substituents on the 1c and 1d derivatives was very high. On the other hand, indazole derivatives generally showed similar inhibitory effects on JAK3 as JAK1, but in this case also, derivatives 1c and 1d substituted with propargyl alcohol showed JAK1 selectivity more than 2 times.

Therefore, the development of a therapeutic agent for rheumatoid arthritis using a selective inhibitor of Janus kinase 1 (JAK1) has been actively conducted worldwide, and the present inventors have found that inhibitors having high selectivity against JAK1 As a result of effort, the inventors have discovered a substance having a JAK1 inhibitory ability which is much higher than that of JAK2 among the indazole derivatives and completed the invention.

As described in detail above, in the present invention, indazole derivatives having a short linear chain substituent such as propargyl alcohol did not inhibit the kinase activity of JAK2, but it was confirmed that JAK1 could have a selective activity inhibitory effect, It can be used as a therapeutic agent for rheumatoid arthritis.

FIG. 1 is a diagram showing the three-dimensional structure of JAK1, JAK2, and JAK3 superimposed on each other, showing the structural differences between JAK1 and other JAK isoenzymes in the glycine-loop.
Figure 2 shows a characteristic cavity formed in the lower part of the substrate binding site of JAK1.
FIG. 3 illustrates the synthesis process of the compounds represented by formula (1).
FIG. 4 illustrates the synthesis of 3-alkynolyl-5-bromo-indazole-7-carboxamides 2a to 2c as main intermediates.
Figure 5 illustrates the synthesis of 4- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) -1H-indazole derivatives 3a - 3b as the main intermediate.
Figure 6 shows the inhibition activity of JAK isozyme activity by indazole derivatives 1a - 1f at a concentration of 10 [mu] M

Hereinafter, the present invention will be described in more detail with reference to Examples. It will be apparent to those skilled in the art that these examples are for illustrative purposes only and that the scope of the invention is not construed as being limited by these examples.

Example  1. 5- Bromo -One- N, N -D- Boc -4-methylbenzene-3-amine ( 9a ) Synthesis of

After dissolving 2,4-dinitrotoluene (1 g, 5.5 mmol) in sulfuric acid (10 mL), NBS (1.17 g, 6.6 mmol) was added over three portions over 45 minutes. The temperature of the reactants during the addition is maintained at 80 ^° C. At the end of the reaction, add ice water and dimethyl chloride to the reaction. The organic layer was washed with a saturated aqueous Na ₂ S ₂ O ₃ solution and concentrated under reduced pressure to obtain a white solid compound 1-bromo-2-methyl-3,5-dinitrobenzene (1.4 g, 98% yield); ¹ H NMR (400 MHz, Acetone- d ₆ )? 8.72 (d, J = 2.3 Hz, 1H), 8.68 (d, J = 8.3 Hz, 1H), 2.66 (s, 3H); Iron powder (642 mg, 19.15 mmol) was added to a mixture of 1-bromo-2-methyl-3,5-dinitrobenzene (1 g, 3.83 mmol) dissolved in ethanol (10 mL) and THF ^o Apply in C. The reaction is stirred at 80 ^° C for 2 hours. The reaction mixture is cooled to room temperature, dichloromethane and water are added, the mixture is filtered through celite, and washed with dichloromethane. Water is added to the cracked organic layer and K ₂ CO ₃ is added until the water layer becomes basic. After that, when the organic layer is separated, water is removed with magnesium sulfate and concentrated under reduced pressure. Purification via silica gel column chromatography afforded a non-separable reduced intermediate (655 mg, 74% yield) as a yellow solid. To a solution of the reduced intermediate (12 mL) dissolved in dichloromethane (655 mg, 2.83 mmol) was added DMAP (1.04 g, 8.49 mmol) and BOC ₂ O (1.85 g, 8.49 mmol). The reaction is stirred at ambient temperature for 3 hours. The resulting mixture was concentrated under reduced pressure and purified by silica gel column chromatography (hexane: dichloromethane: diethyl ether = 100: 100: 1) to give intermediate (989 mg, 81% yield) Obtained as a pale yellow solid. Intermediates in which the amine group is Boc (989 mg, 2.29 mmol) was dissolved in methanol (10 mL), Pd / C (50 mg) was added, and the gas in the flask was removed. Hydrogen gas was added to the reaction flask using a balloon, and the mixture was stirred at room temperature for 4 hours under these conditions. The reaction was filtered through celite, concentrated under reduced pressure and purified by silica gel column chromatography (hexane: ethyl acetate = 5: 1) to give compound 9a (368 mg, 40% yield) in the form of a yellow powder;

^{1 H NMR (400 MHz, Acetone} _ d 6) δ 6.70 (d, J = 2.0 Hz, 1H), 6.56 (d, J = 2.0 Hz, 1H), 4.93 (br s, 2H), 2.24 (s, 3H ), 1.45 (s, 18H).

Example 2: 4-bromo - N, N - di - Boc - 1H - indazol-6-amine sum of 11b St.

5-bromo -1- N, N - di -Boc-4- methylbenzene-3-amine 9a (368 mg, 0.91 mmol) was dissolved in chloroform (3 mL) added to KOAc (94 mg, 0.96 mmol) give. After the temperature of the reaction was lowered to 0 ^° C, acetic anhydride (0.17 mL, 1.82 mmol) was added. 18-crown-6 (48 mg, 0.18 mmol) and isopentyl nitrite (0.27 mL, 2.00 mmol) were added to the reaction mixture and stirred at 65 ^° C for 18 hours . At the end of the reaction, the reaction mixture is cooled to room temperature, and then washed with a saturated aqueous solution of NaHCO ₃ and an aqueous solution of sodium chloride. The residue was purified by silica gel column chromatography (hexane: ethyl acetate = 6: 1) to give 1- (4-bromo-6- (di-Boc) Indazol-1-yl) ethanone 11a (186 mg, 45% yield);

^{1 H NMR (400 MHz, Acetone_} d 6) δ 8.33 (d, J = 0.5 Hz, 1H), 8.23 (s, 1H), 7.56 (d, J = 1.5 Hz, 1H), 2.77 (s, 3H), 1.41 (s, 18H); ¹³ CNMR (100 MHz, Acetone_d ₆ )? 171.7, 152.1, 142.2, 140.1, 139.3, 128.7, 125.8, 114.8, 113.7, 83.6, 28.0, 23.0; Obtained, 6N hydrochloric acid (2 mL) was dissolved in (4-bromo-6- (di-Boc) -1H- indazole -1-yl) -ethanone 11a (186 mg, 0.41 mmol) in methanol (5 mL) to Followed by stirring at room temperature for 2 hours. The methanol was removed by concentration under reduced pressure, dissolved in ethyl acetate, and washed with water. Concentrated under reduced pressure, the organic solvent layer and 4-bromo - N, N - di-1H -Boc- - indazol-6-amine 11 b was obtained (150 mg, 89% yield).

Example  3. 4- Bromo -1H- Indazole  ( 12b ) Synthesis of

To a solution of 3-bromo-2-methylaniline (300 mg, 1.61 mmol) in chloroform (5 mL) was added potassium acetate (166 mg, 1.69 mmol) at room temperature. Acetic anhydride (0.3 mL, 3.22 mmol) was slowly added at 0 ^° C and stirred at room temperature for 15 minutes. When a white solid formed, 18-crown-6 (85 mg, 0.32 mmol) and isopentyl nitrite (0.48 mL, 3.54 Insert the mmol) and stirred under reflux at 65 ^o C for 18 hours after the end of the reaction is cooled to room temperature, and after adding dichloromethane and the reaction mixture was purified by silica gel column chromatography (nucleic acid: ethyl acetate = 5: purification by 1) 12a (189 mg, to obtain a 49% yield) and 12b (127 mg, 40% yield ). compound 12a (189 mg, 0.79 mmol) to at room temperature was added methanol (6 N hydrochloric acid (2 mL) dissolved in a solution in 4 mL) 2 The organic solvent layer was concentrated under reduced pressure to give 4-bromo-1H-indazole ( 12b ) (140 mg, yield 90% yield) as a colorless oil. ).

Example  4. methyl Amino-5- Bromo -3- Mech benzoate  ( 5 ) Synthesis of

Methyl-2-amino-3-methylbenzoate 4 (1 g, 6.1 mmol) was dissolved in dimethylformamide (10 mL) and N -bromosuccinimide (1.1 g, 6.1 mmol) was added. After stirring at room temperature for 6 hours, add ethyl acetate (10 mL) and wash 3 times with 10 mL each with aqueous sodium carbonate solution. The obtained organic layer was treated with magnesium sulfate (MgSO ₄ ) to remove residual water, and then filtered using a filter paper. The mixture was concentrated under reduced pressure, and the resulting concentrate was purified by silica gel column chromatography (hexane: ethyl acetate = 100: 1) to give Compound 5 (928 mg, 62% yield) as a pale gray solid:

^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) 7.88 (d, J = 2.3 Hz, 1H), 7.28 (d, J = 2.3 Hz, 1H), 5.82 (s, 2H), 3.86 (s, 3H ), 2.15 (s, 3H).

Example  5. methyl -5- Bromo - 1H - Indazole -7- Carboxylate  ( 6b ) Synthesis of

Potassium acetate (221 mg, 2.3 mmol) was added to a solution of methyl-2-amino-5-bromo-3-methylbenzoate ( 5 ) (500 mg, 2.2 mmol) in chloroform (5 mL) at room temperature. Crown-6 (116 mg, 0.4 mmol) and isoamyl nitrite (0.6 mL, 4.8 mmol) were added slowly at 0 ^° C and stirred at room temperature for 15 min. Acetic anhydride (0.4 mL, Insert the mmol) and stirred under reflux at 65 ^o C for 18 hours after the end of the reaction is cooled to room temperature, and after adding dichloromethane and the reaction mixture was purified by silica gel column chromatography (nucleic acid: ethyl acetate = 6: purification by 1) 6a (300 mg, to obtain a 46% yield) and 6b (196 mg, 35% yield ). compound 6a (300 mg, 1.0 mmol) and the mixture of methanol (6 N hydrochloric acid (2 mL) dissolved in a solution in 5 mL) 2 at room temperature The organic solvent layer was concentrated under reduced pressure to give 6b (250 mg, 98% yield). The solvent was distilled off under reduced pressure and the residue was dissolved in ethyl acetate and washed with water.

^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) 12.85 (s, 1H), 8.16 (d, J = 1.5 Hz, 1H), 8.13 (t, J = 1.5 Hz, 1H), 8.09 (d, J = 1.5 Hz, 1 H), 4.04 (s, 3 H).

Example  6. tert - Butyl  (5- Bromo - 1H - Indazole -7- Carbonyl ) Carbamate  ( 2a ) Synthesis of

Methyl-5-bromo- 1H -indazole-7-carboxylate (6b) was dissolved (550 mg, 2.2 mmol) in tetrahydrofuran (2 mL) and water (8 mL), was added lithium hydroxide (206 mg, 8.6 mmol) is heated under reflux at 50 ^o C. After 4 h, the pH was adjusted to pH 2 with 2N HCl and the resulting white solid was filtered using a filter paper to give the carboxylic acid form of the material. After dissolving this material in dichloromethane (5 mL), add oxalyl chloride (0.2 mL, 2.2 mmol) at 0 ^° C and stir for 1 hour. The reaction mixture was distilled under reduced pressure, and saturated ammonia (5 mL) dissolved in methanol was added. After stirring for 4 hours, the mixture was distilled under reduced pressure and purified by silica gel column chromatography (nucleic acid: acetone = 2: 1) Intermediate 5-bromo-lH-indazole-7-carboxamide (450 mg, 85% yield) was obtained as a pale brown solid:

^{1 H NMR (400 MHz, MeOD} ) δ (ppm) 11.24 (s, 1H), 10.63 (s, 2H), 8.16 (d, J = 1.5 Hz, 1H), 8.12 (t, J = 1.5 Hz, 1H) , 8.09 (d, J = 1.5 Hz, 1 H). (450 mg, 1.9 mmol) was dissolved in dichloromethane (5 mL), di- tert -butyl dicarbonate (415 mg, 1.9 mmol) and 4- Dimethylaminopyridine (232 mg, 1.9 mmol) was added, and the mixture was stirred at room temperature. After 6 hours, the reaction product was distilled off under reduced pressure and the residue was purified by silica gel column chromatography (nucleic acid: ethyl acetate = 2: 1) to obtain t -butyl- (5-bromo- 1H -indazole-7-carbonyl ) carbamate (2a) (614 mg, was obtained 95% ^{yield): 1 H NMR (400 MHz} , CDCl 3) δ (ppm) 8.18 (d, J = 1.5 Hz, 1H), 8.14 (t, J = 1.5 Hz, 1 H), 8.10 (d, J = 1.5 Hz, 1 H), 1.50 (s, 9H).

Example  7. tert - Butyl - (5- Bromo -3- Iodo - 1H - Indazole -7- Carbonyl ) Carbamate  ( 7 ) Synthesis of

t-butyl (5-bromo-1H-indazol-7-carbonyl) carbamate 2a was dissolved (614 mg, 1.8 mmol) in dimethylformamide (5 mL), N-child ohdoseok god-carboxamide (240 g, 1.8 mmol). After stirring at room temperature for 6 hours, add ethyl acetate (10 mL) and wash 3 times with 10 mL each with aqueous sodium carbonate solution. The obtained organic layer was treated with magnesium sulfate (MgSO ₄ ) to remove residual water, and then filtered using a filter paper. The mixture was concentrated under reduced pressure, and the resulting concentrate was purified by column chromatography on a silica gel column (hexane: ethyl acetate = 2: 1) to obtain the compound t-butyl (5-bromo-3-iodo-1H-indazol- -Carbonyl) carbamate 7 (453 mg, 54% yield) as a pale gray solid:

^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) 12.86 (s, 1H), 10.58 (s, 1H), 8.44 (d, J = 1.5 Hz, 1H), 8.30 (d, J = 1.5 Hz, 1H ), 1.50 (s, 9H).

Example  8. tert - butyl - (5- bromo -3- (3- hydroxyprop -One- yn -One- yl ) -1H- indazole -7- carbonyl ) carbamate  ( 2b ) Wow tert - butyl  (5- bromo -3- (4- hydroxybut -One- yn -One- yl ) -1H- indazole -7- carbonyl ) carbamate  ( 2c ) Synthesis of

7-carbonyl) carbamate 7 (453 mg, 1.0 mmol) was dissolved in dichloromethane, and then tetrakis (triphenylphosphine) palladium (231 mg, 0.2 mmol), copper (I) iodide (95 mg, 0.5 mmol) was added. Propional alcohol or 3-butynyl alcohol (1.0 mmol) and triethylamine (0.3 mL, 2 mmol) were added thereto, followed by stirring at room temperature for 24 hours. Add dichloromethane (10 mL) and wash three times with 10 mL each with aqueous sodium chloride solution. The obtained organic layer was treated with magnesium sulfate (MgSO ₄ ) to remove residual water, and then filtered using a filter paper. The mixture was concentrated under reduced pressure, and the resulting concentrate was purified by silica gel column chromatography (hexane: acetone = 2: 1) to obtain compounds 2b and 2c :

Compound 2b (141 mg, 36% ^{yield) 1 H NMR (400 MHz,} CDCl 3) δ (ppm) 12.85 (s, 1H), 10.58 (s, 1H), 8.40 (d, J = 1.5 Hz, 1H), 8.28 (d, J = 1.5Hz, 1H), 3.62-3.64 (m, 2H), 2.28-2.30 (m, 2H), 1.50 (s, 9H); Compound 2c (122 mg, 30% ^{yield) 1 H NMR (400 MHz,} CDCl 3) δ (ppm) 12.85 (s, 1H), 10.58 (s, 1H), 8.40 (d, J = 1.5 Hz, 1H), 8.28 (d, J = 1.5 Hz, 1H), 4.12-4.16 (m, 2H), 1.49 (s, 9H).

Example  9. Indazole  Synthesis of Derivatives 1a  ~ 1f )

Compound 2 (1 mmol) was dissolved in toluene (10 mL), potassium carbonate (207 mg, 1.5 mmol) was added and compound 3 (1 mmol) and tetrakis (triphenylphosphine) palladium (231 mg, 0.2 mmol ) Is added and heated to reflux at 90 ^° C. After 12 hours, add ethyl acetate (10 mL) to the reaction and wash 3 times with 10 mL each with aqueous sodium chloride solution. The obtained organic layer was treated with magnesium sulfate (MgSO ₄ ) to remove residual water, and then filtered using a filter paper. The mixture was concentrated under reduced pressure, and then dissolved in tetrahydrofuran (10 mL), followed by addition of a 2N aqueous hydrochloric acid solution (5 mL), followed by stirring at room temperature for 4 hours. The reaction is neutralized with aqueous 2N sodium hydroxide solution and extracted three times with 10 mL each with ethyl acetate. The obtained organic layer was treated with magnesium sulfate (MgSO ₄ ) to remove residual water, and then filtered using a filter paper. The mixture was concentrated under reduced pressure, and the resulting concentrate was purified by silica gel column chromatography (dichloromethane: methanol = 20: 1) to obtain Compound 1 :

Compound 1a (188 mg, 68% ^{yield) 1 H NMR (400 MHz,} MeOD) δ (ppm) 12.86 (brs, 2H), 8.14 (d, J = 1.5 Hz, 1H), 8.12 (t, J = 1.5 Hz , 1H), 8.08 (d, J = 1.5 Hz, 1H), 8.05 (s, 1H), 7.61 (d, J = 7.8 Hz, 1H), 7.52 (brs, 2H), 7.32 (d, J = 7.8 Hz , ≪ / RTI > 1H), 7.28 (t, J = 7.8 Hz, 1H); LC / MS (ESI) m / z Found: 278.2 [M + H] < ⁺ >; _{Calcdfor C 15 H 11 N 5 O} : 278.10; Compound 1b (120 mg, 41% ^{yield) 1 H NMR (400 MHz,} MeOD) δ (ppm) 12.86 (brs, 2H), 8.14 (d, J = 1.5 Hz, 1H), 8.12 (t, J = 1.5 Hz , 1H), 8.08 (d, J = 1.5 Hz, 1H), 8.05 (s, 1H), 7.90 (d, J = 1.5 Hz, 1H), 7.52 (brs, 2H), 7.85 (d, J = 1.5 Hz , ≪ / RTI > 1H), 4.95 (brs, 2H); LC / MS (ESI) m / z Found: 293.3 [M + H] < ⁺ >; Calcdfor C ₁₅ H ₁₂ N ₆ O: 293.11; Compound 1c (130 mg, 39% ^{yield) 1 H NMR (400 MHz,} MeOD) δ (ppm) 12.86 (brs, 2H), 8.38 (d, J = 1.5 Hz, 1H), 8.21 (d, J = 1.5 Hz , 1H), 8.04 (s, 1H), 7.60 (d, J = 7.8 Hz, 1H), 7.52 (brs, 2H), 7.32 (d, J = 7.8 Hz, 1H), 7.28 (t, J = 7.8 Hz ), 4.95 (brs, 2H), 4.12-4. 16 (m, 2H); LC / MS (ESI) m / z Found: 332.2 [M + H] < ⁺ >; Calcdfor C ₁₈ H ₁₃ N ₅ O ₂ : 332.11; Compound 1d (128 mg, 37% ^{yield) 1 H NMR (400 MHz,} MeOD) δ (ppm) 12.86 (brs, 2H), 8.38 (d, J = 1.5 Hz, 1H), 8.21 (d, J = 1.5 Hz , 1H), 8.04 (s, 1H), 7.85 (d, J = 1.5 Hz, 1H), 7.79 (d, J = 1.5 Hz, 1H), 7.52 (brs, 2H), 4.95 (brs, 2H), 4.12 -4.16 < / RTI > (m, 2H); LC / MS (ESI) m / z Found: 347.3 [M + H] < ⁺ >; Calcdfor C ₁₈ H ₁₄ N ₆ O ₂ : 347.12; Compound 1e (121 mg, 38% ^{yield) 1 H NMR (400 MHz,} MeOD) δ (ppm) 12.86 (brs, 2H), 8.38 (d, J = 1.5 Hz, 1H), 8.21 (d, J = 1.5 Hz , 1H), 8.04 (s, 1H), 7.59 (d, J = 7.8 Hz, 1H), 7.52 (brs, 2H), 7.31 (d, J = 7.8 Hz, 1H), 7.26 (t, J = 7.8 Hz ), 4.95 (br s, 2H), 4.12-4.16 (m, 2H), 2.26-2.30 (m, 2H); LC / MS (ESI) m / z Found: 346.5 [M + H] < ⁺ >; Calcdfor C ₁₉ H ₁₅ N ₅ O ₂ : 346.12; Compound 1f (132 mg, 37% ^{yield) 1 H NMR (400 MHz,} MeOD) δ (ppm) 12.86 (brs, 2H), 8.38 (d, J = 1.5 Hz, 1H), 8.21 (d, J = 1.5 Hz , 1H), 8.04 (s, 1H), 7.85 (d, J = 1.5 Hz, 1H), 7.79 (d, J = 1.5 Hz, 1H), 7.52 (brs, 2H), 4.95 (brs, 2H), 4.12 -4.16 < / RTI > (m, 2H), 2.26-2.29 (m, 2H); LC / MS (ESI) m / z Found: 362.4 [M + H] < ⁺ >; Calcdfor C ₁₉ H ₁₆ N ₆ O ₂ : 362.13.

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereby. something to do. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

Claims (7)

Claims 1. A compound of the formula 1: < EMI ID =
[Chemical Formula 1]

In Formula 1, n is an integer of 1 or 2, and R 2 is hydrogen or an amino group. The compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein the compound has the formula 1 wherein n is 1 and R 2 is hydrogen or an amino group. delete A pharmaceutical composition for treating or preventing rheumatoid arthritis comprising the compound of claim 1 as an active ingredient. delete A food composition for improving rheumatoid arthritis comprising the compound of claim 1 as an active ingredient. 5-bromo-indazole-7-carboxamide and 4- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan- - Suzuki coupling reaction of an indazole derivative and a Boc-protecting group elimination reaction.

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