KR101965348B1 - N'-hydroxyindazolecarboximidamide derivatives, optical isomer thereof, or pharmaceutically acceptable salts thereof, and, a anticancer composition containing the same as an active ingredient - Google Patents

Abstract

The N'-hydroxyindazole carboxyimidamide derivative compound, its optical isomer or a pharmaceutically acceptable salt thereof, which is a pharmaceutical composition for anticancer medicine according to the present invention, has an inhibitory effect of indoleamine 2,3-dioxygenase-1 And is useful as a pharmaceutical composition for anticancer therapy.

Description

N'-hydroxyindazole carboxyimidamide derivative, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, and an anticancer composition comprising the same as an active ingredient. an anticancer composition containing the same as an active ingredient < RTI ID = 0.0 >

The present invention relates to a N'-hydroxyindazole carboxyimidamide derivative compound, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, and an anticancer composition comprising the compound as an active ingredient.

Cancer is one of the intractable diseases that humanity needs to solve. There is enormous capital invested in development to heal cancer worldwide, medical technology is also developing progressively, but nevertheless cancer deaths continue to increase Trend. According to the National Statistical Office, about 220,000 new cancer patients have been reported as of 2014. These figures are about two times more than the number of new cancer patients in 2002. The number of cancer patients rapidly increases annually, but about 80,000 of the 220,000 cancer patients die from cancer. It is urgent.

Cells, the smallest unit of the body, under normal conditions, divide and grow by the regulatory function of the cells themselves, and die by themselves when they are full or damaged, thus maintaining an overall number balance. However, if there is a problem with the regulation function of the cell itself due to various reasons, abnormal cells that normally should die are over-proliferated. In some cases, they penetrate into surrounding tissues and organs to form a mass, This condition can be defined as cancer.

Complex interactions between cancer cells and immune cells are the most important factors that determine the survival of early cancer cells. Immune cells can detect the genetic modification accumulated in cancer cells and attack cancer cells to prevent the proliferation and metastasis of cancer cells. However, Cancer cells that can evade the attack of cancer can develop into invasive cancer or metastatic cancer.

Recently, the immunosuppressive pathway of cancer has been treated by digesting tryptophan with indoleamine 2,3-dioxygenase (IDO), an immunoregulatory enzyme, to release kynurenine . Tryptophan is an important component of immune evasion, and kininin, the major product of tryptophan degradation, combines with an aryl hydrocarbon receptor (AHR) to evade the immune system and evolve into a progressive cancer. In other words, the IDO inhibits the activation of T cells and other immune cells, thereby preventing the immune system from recognizing and attacking cancer cells. In addition, the IDO is also involved in chronic viral infections and allergies, and is involved in various autoimmune diseases and inflammatory diseases (Muller, AJ et al., Curr Cancer Drug Targets., 7 (1), 31-40, 2007).

It has been reported that IDO is overexpressed in various carcinomas such as breast cancer, blood cancer, kidney cancer, but it is also reported to be involved in cancer cell metastasis (Okamoto, A., et al , Clin Cancer Res., 11 (16), 6030-6039, 2005; Van Baren, N. et al., Front Immunol., 6, 34, 2015, Korean Patent Publication No. 10-2017-0019814).

Cancer Res., 67 (2), 792-801, 2007; Liu, X. et al., &Quot; , Blood., 115 (17), 3520-3530, 2010). These results have demonstrated the possibility of developing an immunotherapeutic agent targeting IDO. In addition, it has been reported that the anti-cancer effect is increased when the IDO inhibitor under development is administered in combination with the existing anti-cancer immunosuppressive antibody, PD-L1 & CTLA antibody (Wainwright, DA et al., Clinical Cancer Res., 20 -5301, 2014).

Human IDO is known as IDO-1 and IDO-2. Among them, IDO-1 is a protein having a molecular weight of 45 kDa consisting of 403 amino acids and is widely expressed in the body but is known to be overexpressed particularly in relation to cancer, chronic inflammatory diseases, allergies, autoimmune diseases and local immunosuppressive diseases . In particular, IDO-1 is closely associated with cancer development, based on studies that IDO-1 is overexpressed in tumors and tumor invading lymph nodes and that tryptophan metabolism is increased in the urine and blood of tumor patients .

On the other hand, Korean Patent Publication No. 10-2015-0087400 discloses a compound useful as an inhibitor of indoleamine 2,3-dioxygenase as a prior art related to anti-cancer composition having inhibitory activity of IDO-1, Patent No. 10-2017-0019814 discloses a novel IDO-1 inhibitor and its use, and as a prior art related to N'-hydroxyindazole carboxyimidamide derivative compounds, International Publication No. 2007087443 discloses a novel IDO- The prior literature disclosing that the vinyl-phenyl derivative for the related use is disclosed but can be used as an anti-cancer composition from the IDO-1 inhibitory effect of the N'-hydroxyindazole carboxyimidamide derivative compound having the structure of the present invention .

Korean Patent Publication No. 10-2015-0087400 (inhibitor of indoleamine 2,3-dioxygenase, published on Jul. 29, 2015) Korean Patent Publication No. 10-2017-0019814 (Novel IDO-1 inhibitor and its use, published on Feb. 22, 2017) International Publication No. 2007087443 (published by VINYL-PHENYL DERIVATIVES FOR INFLAMMATION AND IMMUNE-RELATED USES, Jan. 25, 2007)

Hou, D. Y. et al., Inhibition of indoleamine 2,3-dioxygenase in dendritic cells by stereoisomers of 1-methyl-tryptophan correlates with antitumor responses, Cancer Res., 67 (2), 792-801, 2007. Liu, X. et al., Selective inhibition of IDO1 effectively regulates mediators of antitumor immunity, Blood., 115 (17), 3520-3530, 2010. Muller, A. J. et al., Indoleamine 2,3-dioxygenase in immune suppression and cancer, Curr Cancer Drug Targets., 7 (1), 31-40, 2007. Okamoto, A. et al., Indoleamine 2,3-dioxygenase serving as a marker of poor prognosis in gene expression profiles of serous ovarian cancer cells, Clin Cancer Res., 11 (16), 6030-6039, 2005. Van Baren, N. et al., Tumor immune resistance in Tryptophan-degrading enzymes, Front Immunol., 6, 34, 2015. Wainwright, D. A. et al., Durable therapeutic efficacy using combinatorial blockade against IDO, CTLA-4, and PD-L1 in mice with brain tumors, Clin Cancer Res., 20 (20), 5290-5301, 2014.

It is an object of the present invention to provide an N'-hydroxyindazole carboxyimidamide derivative compound, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, and an anticancer composition comprising the compound as an active ingredient.

The present invention relates to an N'-hydroxyindazole carboxyimidamide derivative represented by the following formula (1), an optical isomer thereof or a pharmaceutically acceptable salt thereof.

[Chemical Formula 1]

In Formula 1,

R ₁ to R ₄ are each independently one or more hydrogen, halogen, hydroxy, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₄ -C ₁₀ cycloalkyl, C ₄ -C ₁₀ aryl, C ₄ - C ₁₀ heterocycloalkyl, C ₄ -C _{10 heteroaryl, -NO 2, -NR '1 R} ' 2, -NR '1 (CO (O) R' 2), -NR '1 (C (O) NR _{'1 R' 2), -COR} '1, -CO (O) R' 1, -C (O) NR '1 R' 2, -CN, -SO (O) R '1, -SO (O) the annular structure of the _{NR '1 R' 2, -NR} '1 (SO (O) R' 2), -CSNR '1 R' 2, or R ₂ and R ₃ are C ₄ -C ₁₀ aryl, by mutual coupling, or Or C ₂ -C ₁₀ heteroaryl, wherein R ' ₁ and R' ₂ are each independently selected from the group consisting of hydrogen, hydroxy, C ₁ -C ₆ alkyl, C ₃ -C ₈ cycloalkyl, C ₄ -C ₁₀ aryl, C ₄ -C ₁₀ aryloxy, C ₁ -C _{8 heteroaryl, - (CR "1 R"} 2) m -C 4 -C 10 _{aryl, - (CR "1 R"} 2) m -C ₄ -C ₁₀ heteroaryl, or NR _"1 R" _2, and wherein R _"1 and R" ₂ are each independently hydrogen, C ₁ -C ₃ Al , Or R _"1 and R" ₂ are coupled together by a C ₄ -C ₁₀ aryl group and a cyclic structure;

m is an integer of 1 to 4;

The term " optical isomer " includes forms of R-form, S-form or racemic compounds, respectively.

The term " alkyl " refers to a straight or branched chain hydrocarbon group of a single bond, for example, methyl, ethyl, propyl, n-butyl, isobutyl, tert-butyl and 1-methylpropyl.

The term " alkoxy " refers to an oxygen group to which a single bond of a saturated or a branched saturated hydrocarbon is bonded, and includes, for example, methoxy, ethoxy, propoxy, n-butoxy, .

The term " cycloalkyl " refers to a saturated, single bond saturated hydrocarbon group, depending on the number of carbon atoms, such as cyclobutyl, cyclopentyl, cyclohexyl, and the like.

The term " aryl " means an aromatic substituent having at least one ring having a covalent pi electron system and includes groups of monocyclic or fused ring polycyclic (i. E., Rings dividing adjacent pairs of carbon atoms) groups .

The term " heterocycloalkyl " refers to a saturated monocyclic saturated hydrocarbon group containing at least one heteroatom such as N, O, or S, and includes, depending on the number and type of heteroatoms contained in the ring, Pyridine, piperidine, piperazine, morpholine, tetrahydrofuran, tetrahydropyran and the like.

The term " heteroaryl " refers to an aromatic ring compound containing at least one heteroatom such as N, O, or S, and may include pyridine, indole, isoindole, and the like depending on the number and type of heteroatoms contained in the ring, .

Examples of the aryl or heteroaryl include, but are not limited to, phenyl, furan, pyran, pyridyl, pyrimidyl, triazyl and the like.

Preferably, in Formula 1,

R ₁ and R ₂ are hydrogen or C ₁ -C ₆ alkyl,

R ₃ and R ₄ are one or more hydrogen, halogen, hydroxy, C ₁ -C ₆ alkyl, C ₁ -C _{6 alkoxy, -NO 2, -NH 2, -COR} '1, -CO (O) R' 1 , -CN, wherein R ' ₁ is hydrogen, hydroxy, C ₁ -C ₆ alkyl, an N, N'-hydroxyindazole carboxyimidamide derivative thereof, an optical isomer thereof or a pharmaceutically acceptable salt thereof Acceptable salt.

More specifically, the compound of formula (1)

(Z) -N- (3-bromo-4-fluorophenyl) -N'-hydroxy-1H-indazole-7-carboximidamide (Compound 1);

(Z) -N- (3-chloro-4-fluorophenyl) -N'-hydroxy-1H-indazole-7-carboximidamide (Compound 2);

(Z) -N- (3-chlorophenyl) -N'-hydroxy-1H-indazole-7-carboximidamide (Compound 3);

(Z) -N- (4-fluorophenyl) -N'-hydroxy-1H-indazole-7-carboxyimidamide (Compound 4);

(Z) -N- (2,5-difluorophenyl) -N'-hydroxy-1H-indazole-7-carboximidamide (Compound 5);

(Z) -N- (3-acetylphenyl) -N'-hydroxy-1H-indazole-7-carboxyimidamide (Compound 6);

(Z) -N'-hydroxy-N- (m-tolyl) -1H-indazole-7-carboxyimidamide (Compound 7);

(Z) -N- (4-chloro-3-methylphenyl) -N'-hydroxy-1H-indazole-7-carboximidamide (Compound 8);

(Z) -N- (3-chloro-4-methoxyphenyl) -N'-hydroxy-1H-indazole-7-carboximidamide (Compound 9);

(Z) -N'-hydroxy-N-phenyl-1H-indazole-7-carboximidamide (Compound 10);

(Z) -N- (4-chlorophenyl) -N'-hydroxy-1 H-indazole-7-carboximidamide (Compound 11);

(Z) -N'-hydroxy-N- (4-methoxyphenyl) -1H-indazole-7-carboximidamide (Compound 12);

(Z) -N'-hydroxy-N- (p-tolyl) -1H-indazole-7-carboximidamide (Compound 13);

(Z) -N- (3-bromo-4-fluorophenyl) -N'-hydroxy-1-methyl-1H-indazole-7-carboximidamide (Compound 14); And

(Z) -N- (3-chloro-4-fluorophenyl) -N'-hydroxy-1-methyl-1H-indazole-7-carboximidamide (Compound 15).

The pharmaceutically acceptable salt of the N'-hydroxyindazolecarboxyimidamide derivative of the present invention is preferably a pharmaceutically acceptable salt such as a hydrochloride, a sulfate, a phosphate, a hydrobromide, a hydroiodide, a nitrate, a pyrosulfate, a metaphosphate And salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid and the like, citric acid, oxalic acid, benzoic acid, acetic acid, trifluoroacetic acid, propionic acid, succinic acid, fumaric acid, lactic acid, maleic acid, An addition salt formed by an organic acid such as a sulfonic acid salt, a toluene sulfonic acid salt or a sulfonic acid salt, or a metal salt such as a lithium salt, a sodium salt, a potassium salt, a magnesium salt and a calcium salt.

The pharmaceutical composition according to the present invention can be formulated into a suitable form together with a commonly used pharmaceutically acceptable carrier. &Quot; Pharmaceutically acceptable " refers to a composition that is physiologically acceptable and, when administered to humans, does not normally cause an allergic reaction such as gastrointestinal disorders, dizziness, or the like. In addition, the compositions can be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterilized injection solutions according to a conventional method.

Examples of carriers, excipients and diluents that can be contained in the composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, But are not limited to, cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methyl parahydroxybenzoate, propyl parahydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, it is prepared using diluents or excipients such as fillers, stabilizers, binders, disintegrants, surfactants and the like which are usually used. Solid form preparations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, microcrystalline cellulose, sucrose or lactose, Low-substituted hydroxypropylcellulose, hypromellose, 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, syrups and the like. In addition to water and liquid paraffin which are commonly used simple diluents, 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 non-aqueous solvent and suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As a base for suppositories, witepsol, macrogol, tween 61, cacao paper, laurin, glycerol, gelatin and the like can be used. In order to formulate the formulation for parenteral administration, the N'-hydroxyindazole carboxyimidamide derivative compound of Formula 1 or its pharmaceutically acceptable salt may be sterilized and / or preserved, stabilized, hydrated or emulsified, Such as excipients, adjuvants such as salts and / or buffers for control, and other therapeutically useful substances, to prepare solutions or suspensions in water and prepare them in ampoules or vial unit dosage forms.

The pharmaceutical composition comprising the compound of formula (I) as an active ingredient of the present invention can be administered to mammals such as rats, livestock, and humans in various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection. The dosage will depend on the age, sex, body weight, the particular disease or condition being treated, the severity of the disease or condition, the time of administration, the route of administration, the absorption, distribution and excretion of the drug, It depends on judgment. Dosage determinations based on these factors are within the level of ordinary skill in the art and generally the dosage ranges from 0.01 mg / kg / day to approximately 2000 mg / kg / day. A more preferable dosage is 1 mg / kg / day to 500 mg / kg / day. The administration may be carried out once a day or divided into several doses. The dose is not intended to limit the scope of the invention in any way.

The present invention also relates to an anticancer pharmaceutical composition or a pharmaceutical composition for inhibiting cancer metastasis comprising an N'-hydroxyindazole carboxyimidamide derivative compound, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient will be.

The cancer is also referred to as malignant tumor. It is an abnormally grown mass due to autonomous overgrowth of body tissue. It is a disease that rapidly grows by infiltrating the surrounding tissues, spreads on the body parts, or becomes a life-threatening disease , And includes carcinoma and sarcoma. More specifically, it may include uterine cancer, breast cancer, brain cancer, rectal cancer, colon cancer, kidney cancer, pancreatic cancer, esophageal cancer, lung cancer, gastric cancer, liver cancer and blood cancer.

In yet another aspect,

(Step 1) hydrogenating compound A of Reaction Scheme 1 to prepare Compound B;

(Step 2) halogenating Compound B prepared in Step 1 to prepare Compound C;

(Step 3) cyanating the compound C prepared in the step 2 to prepare a compound D;

(Step 4) Hydroxyamidation of the compound D prepared in the above Step 3 to prepare the compound E;

(Step 5) halogenating Compound E prepared in Step 4 to prepare Compound F; And

(Step 6) Substituting the compound F prepared in Step 5 to prepare a compound of Formula 1; And a process for producing the N'-hydroxyindazole carboxyimide derivative represented by the general formula (1).

[Reaction Scheme 1]

Step 1 is a step of hydrogenating compound A to prepare compound B, which is a step of introducing a substituent of -NO _{2 as} a substituent of -NH ₂ , wherein the hydrogenation reaction is an organic compound having an unsaturated bond Is a hydrogenation or hydrogenation reaction. Metal catalysts such as platinum, palladium, nickel, iron, cobalt and copper are mainly used as the catalyst, but sulfides such as nickel sulfide and molybdenum sulfide can be used, and they can be reacted at room temperature to 200 ° C, Any process that can be manufactured is included in the present invention without limitation.

Step 2 is a step of halogenating compound B prepared in step 1 to prepare compound C, wherein a substituent of -NH ₂ is introduced as a substituent of -I, wherein the halogenation reaction is one or A further halogen is introduced into the organic compound. Fluorination, chlorination, bromination and iodination, depending on the kind of halogen. In the above step, iodination is preferably used. However, any process capable of producing iodide is included in the present invention without limitation.

The step 3 is a step of cyanating the compound C prepared in the step 2 to prepare a compound D, and introducing a substituent of -I as a substituent of -CN, wherein the cyanation reaction is a cyanation reaction Group. The transition metal complex used as a catalyst precursor in the catalytic reaction is mainly prepared by using palladium and nickel complexes, while the aliphatic compound is cyanated by a halogen derivative using an alkali cyanide and the aromatic compound is substituted by a sulfonic acid group substituent. Are included in the present invention without limitation.

Step 4 is a step of introducing a substituent of -CN to -C (= NOH) NH ₂ in the step of preparing compound E by converting hydroxyapamidination of compound D prepared in step 3, Are included in the present invention without limitation.

Step 5 is a step of halogenating Compound E prepared in Step 4 to prepare Compound F, which is a step of introducing a substituent of -NH _{2 as} a substituent of -Cl. In this step, a chlorination reaction is preferably used And any process capable of producing the same is included in the present invention without limitation.

The step 6 is a step of preparing the compound of the formula (1) by substituting the compound F prepared in the step 5 with aniline.

Further, in the process for preparing the compound represented by the formula (1) according to the present invention, more preferred examples include the process for preparing the compounds of the following examples of the present invention. The method for producing the compound shown in the method for producing the compound of the present invention is to be understood as an example of the method for producing the compound represented by the formula (1), and the method for producing the compound is included in the present invention without limitation . Also, it should be understood that the methods presented in the specification of the present invention and manufacturing methods that can be easily modified and modified by those skilled in the art are also included in the scope of the present invention, ≪ / RTI >

The N'-hydroxyindazole carboxyimidamide derivative compound, its optical isomer or a pharmaceutically acceptable salt thereof, which is a pharmaceutical composition for anticancer medicine according to the present invention, has an inhibitory effect of indoleamine 2,3-dioxygenase-1 And is useful as a pharmaceutical composition for anticancer therapy.

Hereinafter, preferred embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein but may be embodied in other forms. Rather, the intention is to provide an exhaustive, complete, and complete disclosure of the principles of the invention to those skilled in the art.

< Example  1. N'- Hydroxyindazole carboxyimidamide  Synthesis of Derivative Compound &gt;

The N'-hydroxyindazole carboxyimidamide derivative compounds 1 to 15 of the present invention were synthesized by referring to the synthesis process of the following Reaction Scheme 2.

[Reaction Scheme 2]

First, 7 g of 7-nitroindazole (3 g) and 10% Pd / C (0.6 g) were mixed in ethanol (10 ml) at room temperature and stirred under H ₂ pressure. The reaction was then filtered through celite and then with ethanol, and the remaining filtrate was concentrated.

Then, 7-aminoindazole (0.3 g, 2.25 mmol) obtained in the above procedure was dissolved in c-hydrochloric acid (0.56 ml) and 98% sulfuric acid (2 ml), and sodium nitrite dissolved in distilled water (2 ml) 0.154 g, 2.23 mmol), and the mixture was stirred at 0 ° C for 30 minutes. Potassium iodide (0.747 g, 4.5 mmol) dissolved in distilled water (2 ml) was then added dropwise to the reaction, followed by stirring at room temperature for 2 hours. Ethyl acetate was added to the reaction mixture, and the mixture was washed successively with distilled water, saturated sodium hydrogencarbonate and brine. The organic layer was separated, dried over magnesium sulfate, and then purified by MPLC.

(Dibenzylideneacetone), Pd ₂ (dba) ₃ (dibenzylideneacetone) and dppf (1,1'-Ferrocenediyl-bis (diphenylphosphine), 55 mg, 0.098 mmol) (20mg, 0.307mmol) and zinc cyanide (86.6mg, 0.737mmol) were dissolved in DMA (Dimethylacetamide, 30ml, degased) and reacted at 120 ° C for 5 hours . Thereafter, the reaction mixture was concentrated, distilled water and ethyl acetate were added, and then only the organic layer was separated. The separated organic layer was washed with 2N ammonium hydroxide and brine, dried over magnesium sulfate, concentrated, and purified by MPLC to obtain indazol-7-carbonitrile.

The indazole-7-carbonitrile (0.1 g, 0.7 mmol) obtained in the above procedure was dissolved in distilled water (1.5 ml) and ethanol (3 ml), and hydroxylammonium chloride (0.073 g, 1.05 mmol) and sodium hydrogencarbonate 0.118 g, 1.4 mmol) was added, followed by reaction at 85 占 폚 for 8 hours. The reaction mixture was added with distilled water and ethyl acetate to separate only the organic layer, washed with brine, dried over magnesium sulfate and concentrated.

(74 mg, 0.42 mmol) obtained in the above procedure was dissolved in distilled water (1 ml), acetic acid (0.5 ml) and 6N HCl (1.26 mmol, 0.21 ml) And stirred at 45 &lt; 0 &gt; C until the reaction was complete. Thereafter, NaCl (0.074 g, 1.26 mmol) was added thereto, stirred at 0 ° C. (ice / water / methanol bath), and sodium nitrite (0.029 g, 0.42 mmol) dissolved in distilled water The reaction was continued for 4 hours. Thereafter, the mixture was stirred in an ice bath for 1.5 hours, then heated to 15 DEG C, and the precipitate was filtered and washed with cold distilled water.

Next, N'-hydroxy-1H-indazole-7-methanimidyl chloride (25 mg, 0.105 mmol) obtained in the above procedure was dissolved in distilled water (1 ml), and 3- -Fluoroaniline (20 [mu] L, 0.105 mmol) was added, followed by stirring for 10 minutes. Then warm sodium hydrogencarbonate (13 mg, 0.157 mmol) dissolved in distilled water (1 ml) was added dropwise to the reaction mixture, followed by stirring at 60 ° C for 3 hours. After 3 hours, the stirred mixture was extracted with ethyl acetate, and the organic layer was washed with brine. The organic layer was dried over magnesium sulfate, concentrated, and purified by MPLC to obtain the compound 1 of the present invention.

The same procedure as in the synthesis of the compound 1 was conducted except that the corresponding amine was used instead of 3-bromo-4-fluoroaniline to obtain the compounds 2 to 15 of the present invention.

Condition Amine Compound 1 3-Bromo-4-fluoroaniline Compound 2 3-chloro-4-fluoroaniline Compound 3 3-chloroaniline Compound 4 4-fluoroaniline Compound 5 2,5-difluoroaniline Compound 6 3-acetyl aniline Compound 7 3-methylaniline Compound 8 4-chloro-3-methylaniline Compound 9 3-chloro-4-methoxyaniline Compound 10 aniline Compound 11 4-chloroaniline Compound 12 4-methoxyaniline Compound 13 4-methylaniline Compound 14 3-Bromo-4-fluoroaniline Compound 15 3-chloro-4-fluoroaniline

< Example  2. N'- Hydroxyindazole carboxyimidamide  Physicochemical Properties of Derivatives>

Example  2-1. (Z) -N- (3- Bromo -4- Fluorophenyl ) -N'-hydroxy-1H- Indazole -7- Car Leucyximidamide

_{^{1 H NMR (300MHz, CDCl 3}} ) δ 12.01 (s, 1H), 8.14 (s, 1H), 7.73 (d, J = 8.0Hz, 1H), 7.48 (s, 1H), 7.20 (d, J = 7.2 Hz, 1H), 7.01 (t , J = 7.7Hz, 1H), 6.95 (dd, J = 5.7, 2.3Hz, 1H), 6.78 (t, J = 8.4Hz, 1H), 6.58-6.43 (m, 1H ).

Example  2-2. (Z) -N- (3- Chloro -4- Fluorophenyl ) -N'-hydroxy-1H- Indazole -7- Car Leucyximidamide

_{^{1 H NMR (300MHz, CDCl 3}} ) δ 11.96 (brs, 1H), 8.15 (s, 1H), 7.74 (d, J = 8.0Hz, 1H), 7.44 (s, 1H), 7.22 (d, J = 7.2 1H), 7.06-6.94 (m, 1H), 6.85-6.75 (m, 2H), 6.53-6.39 (m, 1H).

Example  2-3. (Z) -N- (3- Chlorophenyl ) -N'-hydroxy-1H- Indazole -7- Carboxyimide mid

_{^{1 H NMR (300MHz, CDCl 3}} ) δ 11.91 (s, 1H), 8.16 (s, 1H), 7.74 (d, J = 8.0Hz, 1H), 7.43 (s, 1H), 7.34-7.25 (m, 1H ), 7.08-6.94 (m, 2H), 6.90 (d, J = 7.9 Hz, IH), 6.77 (s, IH), 6.51 (d, J = 7.8 Hz, IH).

Example  2-4. (Z) -N- (4- Fluorophenyl ) -N'-hydroxy-1H- Indazole -7- Carboxyimide Damida

_{^{1 H NMR (300MHz, CDCl 3}} ) δ 11.98 (s, 1H), 8.15 (s, 1H), 7.71 (dd, J = 8.0, 0.7Hz, 1H), 7.43 (s, 1H), 7.21 (dd, J = 7.3, 0.7 Hz, 1H), 7.02-6.95 (m, 1H), 6.82-6.74 (m, 2H), 6.72-6.63 (m, 2H).

Example  2-5. (Z) -N- (2,5- Difluorophenyl ) -N'-hydroxy-1H- Indazole -7- Carboxy Imidamide

_{^{1 H NMR (300MHz, CDCl 3}} ) δ 8.21 (s, 1H), 7.83 (dd, J = 8.1, 0.8Hz, 1H), 7.467.39 (m, 1H), 7.18 (s, 1H), 7.17-7.10 (m, 1H), 7.10-6.99 (m, 1H), 6.66-6.53 (m, 1H), 6.36-6.24 (m, 1H).

Example  2-6. (Z) -N- (3- Acetylphenyl ) -N'-hydroxy-1H- Indazole -7- Carboxyimide mid

^{1 H NMR (300MHz, DMSO)} δ 12.80 (s, 1H), 10.79 (s, 1H), 8.64 (s, 1H), 8.09 (s, 1H), 7.78 (d, J = 7.9Hz, 1H), 7.35 -7.27 (m, 3H), 7.16-7.04 (m, 2H), 6.88 (d, J = 9.5 Hz, 1H), 2.31 (s, 3H).

Example  2-7. (Z) -N'-hydroxy-N- (m- Tolyl ) -1H- Indazole -7- Carboxyimidamide

_{^{1 H NMR (300MHz, CDCl 3}} ) δ 11.80 (s, 1H), 8.17 (s, 1H), 7.74 (dd, J = 8.1, 0.8Hz, 1H), 7.37-7.27 (m, 2H), 7.11-6.95 (m, 2H), 6.79 (d, J = 7.5 Hz, 1H), 6.66 (s, 1H), 6.54 (d, J = 7.9 Hz,

Example  2-8. (Z) -N- (4- Chloro -3- Methylphenyl ) -N'-hydroxy-1H- Indazole -7- Carlsbad Siimidamide

^{1 H NMR (300MHz, DMSO)} δ 12.77 (s, 1H), 10.75 (s, 1H), 8.44 (s, 1H), 8.10 (d, J = 1.3Hz, 1H), 7.78 (d, J = 7.9Hz , 1H), 7.26 (d, J = 6.3Hz, 1H), 7.12-7.03 (m, 1H), 6.99 (d, J = 8.6Hz, 1H), 6.74 (d, J = 2.6Hz, 1H), 6.39 (dd, J = 8.5, 2.7 Hz, 1 H), 2.10 (s, 3H).

Example  2-9. (Z) -N- (3- Chloro -4- Methoxyphenyl ) -N'-hydroxy-1H- Indazole -7- Car Diplopia imidamide

_{^{1 H NMR (300MHz, CDCl 3}} ) δ 12.0 (brs, 1H), 8.13 (s, 1H), 7.70 (d, J = 8.0Hz, 1H), 7.40 (s, 1H), 7.20 (d, J = 7.3 Hz, 1H), 6.98 (t , J = 7.7Hz, 1H), 6.86 (d, J = 2.6Hz, 1H), 6.58 (d, J = 8.8Hz, 1H), 6.49 (dd, J = 8.8, 2.6 Hz, 1 H), 3.78 (s, 3 H).

Example  2-10. (Z) -N'-hydroxy-N-phenyl-1H- Indazole -7- Carboxyimidamide

_{^{1 H NMR (300MHz, CDCl 3}} ) δ 11.59 (brs, 1H), 8.18 (s, 1H), 7.76 (d, J = 8.0Hz, 1H), 7.34 (d, J = 7.2Hz, 2H), 7.16 ( t, J = 7.6 Hz, 2H), 7.08-6.95 (m, 2H), 6.81 (d, J = 7.8 Hz, 2H).

Example  2-11. (Z) -N- (4- Chlorophenyl ) -N'-hydroxy-1H- Indazole -7- Carboxyimide Damida

_{^{1 H NMR (300MHz, CDCl 3}} ) δ 11.71 (brs, 1H), 8.18 (s, 1H), 7.77 (d, J = 8.0Hz, 1H), 7.37-7.25 (m, 3H), 7.10 (d, J = 8.5 Hz, 2H), 7.06 (t, J = 7.7 Hz, 1H), 6.71 (d, J = 8.5 Hz, 2H).

Example  2-12. (Z) -N'-hydroxy-N- (4- Methoxyphenyl ) -1H- Indazole -7- Carboxyimide Damida

_{^{1 H NMR (300MHz, CDCl 3}} ) δ 11.95 (s, 1H), 8.14 (s, 1H), 7.70 (d, J = 7.9Hz, 1H), 7.37 (s, 1H), 7.23 (d, J = 7.1 Hz), 6.97 (t, J = 7.7 Hz, 1H), 6.72 (d, J = 9.0 Hz, 2H), 6.65 (d, J = 8.9 Hz, 2H), 3.72

Example  2-13. (Z) -N'-hydroxy-N- (p- Tolyl ) -1H- Indazole -7- Carboxyimidamide

_{^{1 H NMR (300MHz, CDCl 3}} ) δ 11.53 (s, 1H), 8.17 (s, 1H), 7.75 (d, J = 8.1Hz, 1H), 7.32 (d, J = 6.6Hz, 1H), 7.27 ( s, 1 H), 7.07-6.99 (m, 1 H), 6.97 (d, J = 8.1 Hz, 2H), 6.73 (d, J = 8.3 Hz, 2H), 2.27

Example  2-14. (Z) -N- (3- Bromo -4- Fluorophenyl ) -N'-hydroxy-1- methyl -1H- Get Sol-7-carboxyimidamide

_{^{1 H NMR (300MHz, CDCl 3}} ) δ 8.02 (s, 1H), 7.82 (dd, J = 8.1, 1.0Hz, 1H), 7.60 (br, 1H), 7.35 (dd, J = 7.1, 1.0Hz, 1H ), 7.20-7.11 (m, 1H), 6.82-6.75 (m, 1H), 6.73-6.64 (m, 1H), 6.38-6.29 (m, 1H), 4.09 (d, J = 6.2 Hz, 3H).

Example  2-15. (Z) -N- (3- Chloro -4- Fluorophenyl ) -N'-hydroxy-1- methyl -1H- Get Sol-7-carboxyimidamide

_{^{1 H NMR (300MHz, CDCl 3}} ) δ 8.05 (s, 1H), 7.73 (d, J = 8.0Hz, 1H), 7.44 (s, 1H), 7.21 (d, J = 7.2Hz, 1H), 7.06- 6.96 (m, 1H), 6.85-6.73 (m, 2H), 6.53-6.39 (m, 1H), 4.09 (d, J = 6.2 Hz, 3H).

< Example  3. Identification of antitumor activity>

In order to confirm the inhibition of IDO-1, IDO-1 over-expressing cell lines were prepared and used in the present invention. The inhibitory effects of tryptophan and the inhibition of kinourenin production by the compounds of the present invention were analyzed by liquid chromatography / mass spectrometry.

First, an IDO-1 over-expressing cell line, IDO1-HEK293 cell line, was prepared using HEK293 cells (human embryonic kidney 293 cell) and inoculated at a rate of 1 × 10 ⁴ cells per well in a 96-well plate. Then, the culture was stabilized at 37 캜 for 2 days, and then the culture was removed, and the medium was replaced with a medium containing 1 mM L-tryptophan and the compounds 1 to 15 of the present invention for 24 hours at 37 캜. After 24 hours, 120 μl of the culture obtained in the above procedure and 2 μM of caffeine dissolved in 100% methanol were mixed at a ratio of 1: 9 and centrifuged to recover only the supernatant. Then, the supernatant and distilled water were mixed at a ratio of 1: 9 And 150 [mu] l aliquots were dispensed into vials. The concentrations of tryptophan and quinolenin were analyzed by LC-MS, and the results are shown in Table 2.

Condition Inhibition rate at 10 μM concentration To reduce tryptophan
Inhibitory effect on On the generation of kinurenin
Inhibitory effect on Compound 1 100% 100% Compound 2 78% 84% Compound 3 71% 79% Compound 7 27% 27% Compound 12 25% 25% Compound 15 23% -

Referring to the above Table 2, when the compound of the present invention was treated in an IDO1-HEK293 cell line overexpressing IDO-1, the inhibitory effect on tryptophan reduction and the inhibitory effect on the production of kinourenine could be confirmed.

Accordingly, the N'-hydroxyindazole carboxyimidamide derivative of the present invention is excellent in the effect of inhibiting IDO-1, which degrades tryptophan into kinulinin, and thus can be usefully used as an anticancer composition.

< Formulation example  1. Pharmaceutical preparations>

Formulation example  1-1. Sanje  Produce

2 g of the compound 1 of the present invention and 1 g of lactose were mixed and filled in an airtight container to prepare a powder.

Formulation example  1-2. Manufacture of tablets

100 mg of Compound 1 of the present invention, 100 mg of microcrystalline cellulose, 60 mg of lactose hydrate, 20 mg of low-substituted hydroxypropylcellulose and 2 mg of magnesium stearate were mixed and tableted according to a conventional preparation method .

Formulation example  1-3. Preparation of capsules

100 mg of Compound 1 of the present invention, 100 mg of microcrystalline cellulose, 60 mg of lactose hydrate, 20 mg of low-substituted hydroxypropylcellulose and 2 mg of magnesium stearate were mixed, and the above components were added Mixed and filled into gelatin capsules to prepare capsules.

Formulation example  1-4. Injection preparation

10 mg of the Compound 1 of the present invention was mixed with the sterilized distilled water suitable amount for injection and the pH adjuster in proper amounts, and then the contents of the above components were prepared per ampoule (2 ml) according to the usual injection preparation method.

Claims (8)

An N'-hydroxyindazole carboxyimidamide derivative represented by the following formula (1), an optical isomer thereof or a pharmaceutically acceptable salt thereof:
[Chemical Formula 1]

In Formula 1,
R ₁ , R ₂ and R ₃ are hydrogen or C ₁ -C ₆ alkyl,
R ₄ is one or more hydrogen, halogen, hydroxy, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, -COR _'1 or -CO (O) R' _1,
Wherein R ' ₁ is hydrogen, hydroxy or C ₁ -C ₆ alkyl. delete The method according to claim 1,
The compound of formula (1)
(Z) -N- (3-bromo-4-fluorophenyl) -N'-hydroxy-1H-indazole-7-carboximidamide (Compound 1);
(Z) -N- (3-chloro-4-fluorophenyl) -N'-hydroxy-1H-indazole-7-carboximidamide (Compound 2);
(Z) -N- (3-chlorophenyl) -N'-hydroxy-1H-indazole-7-carboximidamide (Compound 3);
(Z) -N- (4-fluorophenyl) -N'-hydroxy-1H-indazole-7-carboxyimidamide (Compound 4);
(Z) -N- (2,5-difluorophenyl) -N'-hydroxy-1H-indazole-7-carboximidamide (Compound 5);
(Z) -N- (3-acetylphenyl) -N'-hydroxy-1H-indazole-7-carboxyimidamide (Compound 6);
(Z) -N'-hydroxy-N- (m-tolyl) -1H-indazole-7-carboxyimidamide (Compound 7);
(Z) -N- (4-chloro-3-methylphenyl) -N'-hydroxy-1H-indazole-7-carboximidamide (Compound 8);
(Z) -N- (3-chloro-4-methoxyphenyl) -N'-hydroxy-1H-indazole-7-carboximidamide (Compound 9);
(Z) -N'-hydroxy-N-phenyl-1H-indazole-7-carboximidamide (Compound 10);
(Z) -N- (4-chlorophenyl) -N'-hydroxy-1 H-indazole-7-carboximidamide (Compound 11);
(Z) -N'-hydroxy-N- (4-methoxyphenyl) -1H-indazole-7-carboximidamide (Compound 12);
(Z) -N'-hydroxy-N- (p-tolyl) -1H-indazole-7-carboximidamide (Compound 13);
(Z) -N- (3-bromo-4-fluorophenyl) -N'-hydroxy-1-methyl-1H-indazole-7-carboximidamide (Compound 14); And
(Z) -N- (3-chloro-4-fluorophenyl) -N'-hydroxy-1-methyl-1H-indazole-7-carboximidamide (Compound 15); And the optically active isomer thereof, or a pharmaceutically acceptable salt thereof, wherein the optically active isomer thereof is selected from the group consisting of N'-hydroxyindazole carboxyimidamide derivative, its optical isomer or a pharmaceutically acceptable salt thereof. An anticancer pharmaceutical composition comprising the N'-hydroxyindazole carboxyimidamide derivative compound of claim 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient. 5. The method of claim 4,
Wherein the cancer is selected from the group consisting of cervical cancer, breast cancer, brain cancer, rectal cancer, colon cancer, renal cancer, pancreatic cancer, esophageal cancer, lung cancer, gastric cancer, liver cancer and blood cancer. A pharmaceutical composition for inhibiting cancer metastasis comprising the N'-hydroxyindazole carboxyimidamide derivative compound of claim 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient. The method according to claim 6,
Wherein said cancer is selected from the group consisting of cervical cancer, breast cancer, brain cancer, rectal cancer, colon cancer, renal cancer, pancreatic cancer, esophageal cancer, lung cancer, gastric cancer, liver cancer and blood cancer. (Step 1) hydrogenating compound A of Reaction Scheme 1 to prepare Compound B;
(Step 2) halogenating Compound B prepared in Step 1 to prepare Compound C;
(Step 3) cyanating the compound C prepared in the step 2 to prepare a compound D;
(Step 4) Hydroxyamidation of the compound D prepared in the above Step 3 to prepare the compound E;
(Step 5) halogenating Compound E prepared in Step 4 to prepare Compound F; And
(Step 6) Substituting the compound F prepared in Step 5 to prepare a compound of Formula 1; 2. A process for preparing N'-hydroxyindazole carboxyimidamide derivative represented by the general formula (1) according to claim 1, wherein R &lt; 1 &gt;
[Reaction Scheme 1]