Ahr modulator compound with improved solubility

Abstract

The present application relates to an AhR modulator compound with improved solubility.

Description

AhR modulator compounds with improved solubility

[0001] The present application relates to an AhR modulator compound with improved solubility.

[0002] The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor and is well-known as a crucial intracellular chemical sensor that responds to both natural and artificial environmental compounds. Several studies have demonstrated that AhR signaling plays a crucial role in various diseases, including autoimmunity, infection, and cancer. AhR can influence cancer cell proliferation, tissue invasion, metastasis, and angiogenesis, and is known to be associated with autoimmune diseases, including rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), and multiple sclerosis (MS). Therefore, the development of AhR-targeted therapies could offer a potential opportunity to overcome immune-related diseases.

[0003] Although the development and use of various pharmaceuticals significantly contribute to human health, many compounds have inherently low solubility, significantly reducing their absorption in the body. This low solubility reduces drug bioavailability, necessitating higher doses to achieve effective therapeutic concentrations, which can increase the risk of adverse effects. Previous studies have attempted various approaches, but methods utilizing nanoparticles, for example, have limitations such as complex and expensive manufacturing processes. Furthermore, these methods can potentially compromise drug stability and efficacy.

[0004] Therefore, to solve the problem of low bioavailability due to low solubility of AhR modulator compounds, AhR modulator compounds with improved solubility are required.

[0005] To address these issues, the present inventors adopted a prodrug approach to improve the bioavailability of drugs with low solubility. A prodrug is a precursor that is pharmacologically inactive or has low activity in vivo and is converted into an active drug through metabolic processes. A compound in the form of a prodrug according to one embodiment of the present invention improves the solubility of the drug, thereby increasing its absorption rate in the body, thereby enabling sufficient therapeutic effects to be achieved even at lower doses.

[0006] One example of this application is expected to contribute to improving the effectiveness of drug therapy and reducing the risk of side effects by addressing drug solubility and bioavailability issues. This is expected to maximize drug safety and efficacy, thereby increasing patient satisfaction with treatment.

[0007] One example of the present application relates to an AhR modulator compound having improved solubility, an enantiomer, a diastereomer, a racemate, a solvate, a hydrate, or a pharmaceutically acceptable salt thereof.

[0008] Another example of the present application relates to a pharmaceutical composition comprising a compound according to an example of the present application, an enantiomer, a diastereomer, a racemate, a solvate, a hydrate, or a pharmaceutically acceptable salt thereof.

[0009] Another example of the present application relates to a composition for modulating AhR activity, comprising a compound according to an example of the present application, an enantiomer, a diastereomer, a racemate, a solvate, a hydrate, or a pharmaceutically acceptable salt thereof.

[0010] Another example of the present application relates to a method for preventing or treating a disease mediated by AhR, comprising administering a compound according to an example of the present application, or an enantiomer, diastereomer, racemate, solvate, hydrate, or pharmaceutically acceptable salt thereof.

[0011] Another example of the present application relates to a method of modulating AhR activity, comprising administering a compound according to an example of the present application, or an enantiomer, diastereomer, racemate, solvate, hydrate, or pharmaceutically acceptable salt thereof.

[0012] Another example of the present application relates to a method for inhibiting proliferation, tissue invasion, metastasis and / or angiogenesis of cancer cells in a subject having cancer or a malignant tumor, comprising administering a compound according to an example of the present application, an enantiomer, a diastereomer, a racemate, a solvate, a hydrate, or a pharmaceutically acceptable salt thereof.

[0013] Another example of the present application relates to the use of a compound according to an example of the present application, or an enantiomer, diastereomer, racemate, solvate, hydrate, or pharmaceutically acceptable salt thereof, for modulating AhR activity.

[0014] Another example of the present application relates to the use of a compound according to an example of the present application, or an enantiomer, diastereomer, racemate, solvate, hydrate, or pharmaceutically acceptable salt thereof, for the prevention or treatment of a disease mediated by AhR.

[0015] Another example of the present application relates to the use of a compound according to an example of the present application, or an enantiomer, diastereomer, racemate, solvate, hydrate, or pharmaceutically acceptable salt thereof, for inhibiting proliferation, tissue invasion, metastasis, and / or angiogenesis of cancer cells in a subject having cancer or a malignant tumor.

[0016]

[0017] Hereinafter, the present application will be described in more detail.

[0018] Unless otherwise defined, all technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains. Furthermore, although this application has been described with respect to specific methods and samples, analogs or equivalents thereof should fall within the scope of this application. Furthermore, unless explicitly stated otherwise, numerical values ​​described herein are to be considered to include the meaning of "about." All publications and other references mentioned herein are incorporated herein by reference in their entirety.

[0019]

[0020] compound

[0021] An example of the present application relates to a compound of formula (I), an enantiomer, a diastereomer, a racemate, a solvate, a hydrate, or a pharmaceutically acceptable salt thereof:

[0022] [Chemical formula (I)]

[0023]

[0024] In the above chemical formula (I),

[0025] X is A or -LA,

[0026] L is C 1-5 It is alkyl,

[0027] A is -O(P=O)(OH)2, -O(C=O)CH3, -OSO2CH3, -O(C=O)-pyridinyl, or -O(C=O)-C 1-4 Alkyl-phenyl, and Y is halogen.

[0028]

[0029] For example, the above X may be -LA.

[0030] For example, the above L is C1-3 It could be an alkyl.

[0031] For example, the above L may be ethyl.

[0032] For example, the above A is -O(P=O)(OH)2, -O(C=O)CH3, -O(C=O)-pyridinyl, or -O(C=O)-C 1-4 It may be alkyl-phenyl.

[0033] For example, the above A may be -O(P=O)(OH)2.

[0034] For example, the above Y may be Cl.

[0035] For example, a compound or a hydrate thereof according to an example of the present application may be selected from the following (1) to (6):

[0036] (1) 2-((4-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperazin-1-yl)sulfonyl)ethyl dihydrogen phosphate;

[0037] (2) 2-((4-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperazin-1-yl)sulfonyl)ethyl dihydrogen phosphate dihydrate;

[0038] (3) 2-((4-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperazin-1-yl)sulfonyl)ethyl acetate;

[0039] (4) 2-((4-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperazin-1-yl)sulfonyl)ethyl methanesulfonate;

[0040] (5) 2-((4-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperazin-1-yl)sulfonyl)ethyl nicotinate; and

[0041] (6) 2-((4-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperazin-1-yl)sulfonyl)ethyl 4-phenylbutanoate.

[0042] Single stereochemically isomers, enantiomers, diastereomers, and pharmaceutically acceptable salts of the compounds exemplified above are also within the scope of the present application. Pharmaceutically acceptable salts may be derived, for example, from suitable inorganic and organic acids and bases.

[0043] Acid addition salts can be prepared by reacting the purified compound in free-base form with a suitable organic or inorganic acid, if available, and isolating the salt thus formed. Examples of pharmaceutically acceptable acid addition salts include, without limitation, salts of amino groups formed with inorganic acids such as hydrochloric, hydrobromic, phosphoric, sulfuric, and perchloric acids, or with organic acids such as acetic, oxalic, maleic, tartaric, citric, succinic, or malonic acids.

[0044] Base addition salts can be prepared by reacting a purified compound in acid form with a suitable organic or inorganic base and isolating the salt thus formed. Such salts include alkali metals (e.g., sodium, lithium, and potassium), alkaline earth metals (e.g., magnesium and calcium), ammonium, and N + (C 1-4 Includes, without limitation, alkyl)4 salts.

[0045] Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, glycolate, gluconate, glycolate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, Contains oleate, oxalate, palmitate, palmoate, pectate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, salicylate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate and valerate salts.

[0046] Furthermore, compounds according to an example of the present application include, but are not limited to, their pharmaceutically acceptable salts, as well as all solvates or hydrates and all possible stereoisomers that can be prepared therefrom. All stereoisomers of the compounds of the present application (e.g., those that can exist due to asymmetric carbons on various substituents), including enantiomeric forms and diastereomeric forms, are contemplated within the scope of the present application. Individual stereoisomers of the compounds of the present application may, for example, be substantially free of other isomers (e.g., pure or substantially pure, an optical isomer having a particular activity), or may be, for example, a racemate, or may be a mixture with all other stereoisomers, or with selected other stereoisomers. The chiral centers of the compounds according to the present application may have the S or R configuration as defined by the IUPAC 1974 Recommendation. Racemic forms can be analyzed by physical methods such as fractional crystallization, separation of diastereomeric derivatives, or separation by crystallization or chiral column chromatography. Individual optical isomers can be obtained from the racemate by any suitable method, including but not limited to salt formation using an optically active acid followed by crystallization.

[0047]

[0048] Solubility, conversion, and bioavailability

[0049] The compound according to an example of the present application, its enantiomer, diastereomer, racemate, solvate, hydrate, or pharmaceutically acceptable salt may have improved solubility. Specifically, the compound according to an example of the present application may have improved solubility compared to the compound of Preparation Example 1. As a result of analyzing the solubility of the compound according to an example of the present application in the examples herein, it exhibited significantly higher solubility in water (e.g., water having a pH of 6 to 7, e.g., distilled water) or potassium phosphate compared to the compound of Preparation Example 1 (2-((4-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperazin-1-yl)sulfonyl)ethan-1-ol). Specifically, the terminal hydroxyl group of the compound of Preparation Example 1 is -O(P=O)(OH)2, -O(C=O)CH3, -OSO2CH3, -O(C=O)-pyridinyl, or -O(C=O)-C 1-4 As a result of substitution with alkyl-phenyl, solubility was improved. In particular, when the terminal hydroxyl group of the compound of Preparation Example 1 was substituted with -O(P=O)(OH)2, solubility was significantly improved. Accordingly, in the above chemical formula (I), A may be -O(P=O)(OH)2.

[0050] The compound, its enantiomer, diastereomer, racemate, solvate, hydrate, or pharmaceutically acceptable salt according to an example of the present application may have a solubility in water (e.g., water having a pH of 6 to 7, e.g., distilled water) of at least 10 uM, at least 15 uM, at least 20 uM, at least 25 uM, at least 30 uM, at least 35 uM, at least 40 uM, at least 45 uM, at least 50 uM, at least 55 uM, at least 60 uM, at least 65 uM, at least 70 uM, at least 75 uM, at least 80 uM, at least 85 uM, at least 90 uM, at least 95 uM, or at least 100 uM.

[0051] The compound, its enantiomer, diastereomer, racemate, solvate, hydrate, or pharmaceutically acceptable salt according to an example of the present application may have an improved conversion rate into an active drug. Specifically, the compound according to an example of the present application may have an improved conversion rate in vivo into the compound of Preparation Example 1. As a result of analyzing the bioconversion rate of the compound according to an example of the present application in the examples herein, the conversion rate into an active drug was superior compared to the compounds of Comparative Examples 1 and 2 having a similar parent nucleus structure. Specifically, the terminal hydroxyl group of the compound of Preparation Example 1 is -O(P=O)(OH)2, -O(C=O)CH3, -O(C=O)-pyridinyl, or -O(C=O)-C 1-4 When substituted with alkyl-phenyl, the conversion rate to an active drug was excellent. In particular, when the terminal hydroxyl group of the compound of Preparation Example 1 was substituted with -O(P=O)(OH)2, the conversion rate to an active drug was significantly excellent.

[0052] The conversion rate into the above active drug may be a specific effect of the compound according to an example of the present application. Specifically, the compounds of Comparative Examples 1 and 2 are compounds in which the terminal hydroxyl group of the compounds of Preparation Examples 2 and 3 is substituted with a dihydrogen phosphate group, respectively, and are prodrugs of the same type as the compounds of Examples 1 and 2 in which the terminal hydroxyl group of the compound of Preparation Example 1 is substituted with a dihydrogen phosphate group. Although the compounds of Preparation Examples 2 and 3 have a similar structure to the compound of Preparation Example 1, the compound of Example 1 exhibited a significantly superior bioconversion rate compared to those of Comparative Examples 1 and 2. This means that the effect of improving the conversion rate by substituting the terminal hydroxyl group with a dihydrogen phosphate group is a specific effect of the compound of Example 1.

[0053] The compound according to an example of the present application, its enantiomer, diastereomer, racemate, solvate, hydrate, or pharmaceutically acceptable salt may have improved bioavailability. As described above, the compound according to an example of the present application has high solubility and conversion rate, so when administered in vivo, it has a high absorption rate and is rapidly converted into an active drug, thereby exhibiting excellent bioavailability.

[0054] Accordingly, another example of the present application is -(P=O)(OH)2, -(C=O)CH3, -SO2CH3, -(C=O)-pyridinyl, and -(C=O)-C at the terminal hydroxyl group of the compound of the following formula (II). 1-4 A method for increasing the solubility of a compound of the following formula (II), comprising the step of introducing a functional group selected from the group consisting of alkyl-phenyl:

[0055] [Chemical formula (II)]

[0056]

[0057] In the above chemical formula (II),

[0058] LL is a single bond or C 1-5 It is alkyl,

[0059] YY is a halogen.

[0060]

[0061] For example, the functional group may be -(P=O)(OH)2.

[0062] For example, the above LL is C 1-3 It could be an alkyl.

[0063] Another example of the present application relates to a method for increasing the solubility of a compound of formula (II), comprising a step of phosphorylating the compound of formula (II). The humanization may be phosphorylating a terminal hydroxyl group of the compound of formula (II).

[0064]

[0065] Pharmaceutical compositions and uses

[0066] Another example of the present application relates to a pharmaceutical composition comprising a compound according to an example of the present application, an enantiomer, a diastereomer, a racemate, a solvate, a hydrate, or a pharmaceutically acceptable salt thereof.

[0067] The above pharmaceutical composition may be used for the prevention or treatment of a disease mediated by aryl hydrocarbon receptor (AhR).

[0068] The above pharmaceutical composition may be used to inhibit proliferation, tissue invasion, metastasis and / or angiogenesis of cancer cells in a subject having cancer or a malignant tumor.

[0069] Another example of the present application relates to a composition for modulating AhR activity, comprising a compound according to an example of the present application, an enantiomer, a diastereomer, a racemate, a solvate, a hydrate, or a pharmaceutically acceptable salt thereof. For example, the modulation of AhR activity may be AhR inhibition.

[0070] Another example of the present application relates to a method for preventing or treating a disease mediated by an aryl hydrocarbon receptor (AhR), comprising administering to a subject in need thereof a compound according to an example of the present application, or an enantiomer, diastereomer, racemate, solvate, hydrate, or pharmaceutically acceptable salt thereof.

[0071] Another example of the present application relates to a method for modulating AhR activity, comprising administering to a subject in need thereof a compound according to an example of the present application, or an enantiomer, diastereomer, racemate, solvate, hydrate, or pharmaceutically acceptable salt thereof. For example, the method for modulating AhR activity may be a method for inhibiting AhR.

[0072] Diseases mediated by the above aryl hydrocarbon receptor (AhR) may be disorders associated with abnormal AhR signaling.

[0073] The disease mediated by the above aryl hydrocarbon receptor (AhR) may be a disease mediated by activation of the aryl hydrocarbon receptor (AhR).

[0074] Diseases mediated by the above aryl hydrocarbon receptor (AhR) may be cancer, malignancies, fibrotic disorders, or conditions with dysregulated immune responses.

[0075] The cancer may be selected from the group consisting of breast cancer, squamous cell carcinoma, lung cancer, peritoneal cancer, hepatocellular carcinoma, gastric cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, colon cancer, large intestine cancer, endometrial cancer or uterine cancer, salivary gland carcinoma, kidney or renal cancer, prostate cancer, vulvar cancer, thyroid cancer, head and neck cancer, B-cell lymphoma, chronic lymphocytic leukemia (CLL); acute lymphoblastic leukemia (ALL), hairy cell leukemia, and chronic myeloblastic leukemia.

[0076] Examples of breast cancer include, but are not limited to, triple negative breast cancer, invasive ductal carcinoma, invasive lobular carcinoma, ductal carcinoma in situ, and lobular carcinoma in situ.

[0077] Examples of lung cancers include, but are not limited to, small cell and non-small cell lung carcinomas, as well as bronchial adenomas and pleuropulmonary blastomas.

[0078] Examples of brain cancers include, but are not limited to, neuroectodermal and pineal tumors, as well as brainstem and hypothalamic gliomas, cerebellar and cerebral astrocytomas, glioblastomas, medulloblastomas, and ependymomas.

[0079] Tumors of the male reproductive organs include, but are not limited to, prostate cancer and testicular cancer.

[0080] Tumors of the female reproductive organs include, but are not limited to, uterine sarcoma, as well as endometrial cancer, cervical cancer, ovarian cancer, vaginal cancer, and vulvar cancer.

[0081] Examples of ovarian cancer include, but are not limited to, serous tumors, endometrioid tumors, mucinous cystadenocarcinomas, granulosa cell tumors, Sertoli-Leydig cell tumors, and arrhenoblastomas.

[0082] Examples of cervical cancer include, but are not limited to, squamous cell carcinoma, adenocarcinoma, adenosquamous carcinoma, small cell carcinoma, neuroendocrine tumors, hyaline cell carcinoma, and villous adenocarcinoma.

[0083] Gastrointestinal tumors include, but are not limited to, anal cancer, colon cancer, colorectal cancer, esophageal cancer, gallbladder cancer, stomach cancer, pancreatic cancer, rectal cancer, small intestine cancer, and salivary gland cancer.

[0084] Examples of esophageal cancers include, but are not limited to, squamous cell carcinoma, leiomyosarcoma, malignant melanoma, rhabdomyosarcoma, and lymphoma, as well as esophageal cell carcinoma and adenocarcinoma.

[0085] Examples of gastric cancer include, but are not limited to, intestinal type and diffuse type gastric adenocarcinoma.

[0086] Examples of pancreatic cancer include, but are not limited to, ductal adenocarcinoma, adenosquamous carcinoma, and pancreatic endocrine tumors.

[0087] Urologic tumors include, but are not limited to, bladder, penile, kidney, renal pelvis, ureter, urethra, and human papillary renal cell carcinoma.

[0088] Examples of kidney cancers include, but are not limited to, renal cell carcinoma, urothelial carcinoma, juxtamembrane cell tumor (reninoma), angiomyolipoma, renal oncocytoma, Bellini duct carcinoma, clear cell sarcoma of the kidney, mesoblastic nephroma, and Wilms' tumor.

[0089] Examples of bladder cancer include, but are not limited to, transitional cell carcinoma, squamous cell carcinoma, adenocarcinoma, sarcoma, and small cell carcinoma.

[0090] Ocular cancers include, but are not limited to, intraocular melanoma and retinoblastoma.

[0091] Examples of liver cancer include, but are not limited to, hepatocellular carcinoma (liver cell carcinoma with or without fibrous layer mutation), cholangiocarcinoma (intrahepatic cholangiocarcinoma), and mixed hepatocellular cholangiocarcinoma.

[0092] Skin cancers include, but are not limited to, squamous cell carcinoma, Kaposi's sarcoma, malignant melanoma, Merkel cell skin cancer, and non-melanoma skin cancer.

[0093] Head and neck cancers include, but are not limited to, head and neck squamous cell carcinoma, laryngeal cancer, hypopharyngeal cancer, nasopharyngeal cancer, oropharyngeal cancer, salivary gland cancer, lip and oral cavity cancer, and squamous cell.

[0094] Lymphomas include, but are not limited to, AIDS-related lymphoma, non-Hodgkin's lymphoma, cutaneous T-cell lymphoma, Burkitt lymphoma, Hodgkin's disease, and central nervous system lymphoma.

[0095] Sarcomas include, but are not limited to, soft tissue sarcoma, osteosarcoma, malignant fibrous histiocytoma, lymphosarcoma, and rhabdomyosarcoma.

[0096] Leukemia includes, but is not limited to, acute myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, and hairy cell leukemia.

[0097] The subject may be a mammal, including a human or mammalian cell; for example, a mammal (e.g., a human) or a mammalian cell isolated therefrom suffering from a disease, disorder, or condition associated with AhR activity as described above.

[0098] The compound or pharmaceutical composition as an active ingredient may be administered orally or parenterally. For example, parenteral administration may be performed by any one of intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, intradermal administration, topical administration, intranasal administration, intrapulmonary administration, and rectal administration.

[0099] The above effective amount may refer to a pharmaceutically and / or therapeutically effective amount, and may be prescribed depending on factors such as the type of preparation (formulation), route of administration, age, weight, sex and / or pathological condition of the patient.

[0100] A pharmaceutical composition according to an example of the present application may be formulated into an appropriate form with a commonly used pharmaceutically acceptable carrier. The term "pharmaceutically acceptable" means physiologically tolerable and does not generally cause allergic reactions or similar reactions such as gastrointestinal upset and dizziness when administered to humans. In addition, the pharmaceutical composition of the present application may be formulated and used as oral preparations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, and as parenteral preparations such as topical preparations, suppositories, or sterile injectable solutions, according to conventional methods.

[0101] Examples of carriers, excipients, and diluents that may be included in the composition include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. When formulated into a preparation, diluents or excipients such as commonly used fillers, stabilizers, binders, disintegrants, and surfactants may be used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations can be prepared by mixing the compound of the present application with at least one excipient, such as starch, microcrystalline cellulose, sucrose, lactose, low-substituted hydroxypropyl cellulose, hypromellose, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, oral solutions, emulsions, syrups, etc. In addition to simple diluents such as commonly used water and liquid paraffin, various excipients such as wetting agents, sweeteners, flavoring agents, and preservatives can also be included. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solutions or suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases may include withepsol, macrogol, tween 61, cocoa butter, lauric butter, and glycerogelatin.To be formulated as a parenteral preparation, the compound of formula I or a pharmaceutically acceptable salt thereof may be mixed in water in a sterile state, and / or may include adjuvants such as preservatives, stabilizers, wet powders or emulsifying agents, salts for controlling osmotic pressure, and / or buffers, and other therapeutically useful substances, and then prepared as a solution or suspension and then prepared in the form of unit dosage forms in ampoules or vials.

[0102] As used herein, the terms "treat," "treating," and "treatment" refer to a method of alleviating or terminating a biological disorder and / or at least one of its accompanying symptoms. They are used in their ordinary sense, such as managing or caring for a subject for the purpose of eradicating, alleviating, reducing, alleviating, or improving the condition of a disease or disorder, such as cancer. As used herein, "alleviating" a disease, disorder, or condition means reducing the severity and / or frequency of the symptoms of the disease, disorder, or condition. Furthermore, references to "treatment" herein include references to curative, palliative, and preventative treatments. Treatment of cancer includes inhibiting cancer growth (including causing partial or complete regression of the cancer), inhibiting cancer progression or metastasis, preventing cancer recurrence or residual disease, and / or prolonging the survival of a patient. A "therapeutically effective amount" is an amount of a drug that can achieve the desired therapeutic, palliative, or preventative effect for the condition being treated.

[0103]

[0104] Effective amount

[0105] In some embodiments, the effective dose range of a compound may be determined by measuring the patient's blood compound concentration under a specific dosing regimen to establish a concentration-time profile, consulting the established correlation between the concentration-time profile obtained during the trial and the tumor inhibition or elimination, and further considering the patient's health status or physical endurance, thereby balancing the achievable therapeutic effect with the potential for toxicity to the patient. The frequency of compound administration may be determined similarly. The administration may continue until the patient is cancer-free.

[0106] In some embodiments, the effective dose for tumor treatment may be determined by the patient's ability to stabilize disease progression and / or improve symptoms, and preferably reverse disease progression (e.g., by reducing tumor size). In some embodiments, a maintenance dose may be administered to ensure complete removal or eradication of the cancer, or prevention of residual disease, once the patient is cancer-free. The duration of maintenance administration may be determined based on clinical trial data.

[0107] In some embodiments, the compound may be administered in combination with one or more other cancer therapeutics that target AhR or molecules other than AhR. The compound may be formulated separately or together with the other cancer therapeutics. The compound may be administered on the same or a different schedule as the other cancer therapeutics. The ratio of the compound to the other cancer therapeutics may be determined through clinical trials. Combining the compound with the other cancer therapeutics may further enhance the efficacy of each other. For example, the compound of the present application may be administered with an immune checkpoint inhibitor, such as an inhibitor of PD-1, PD-L1, or PD-L2 (e.g., pembrolizumab, nivolumab, or atezolizumab), or with CAR-T therapy (e.g., axicabtagene ciloleucel), to achieve additive or synergistic anticancer effects.

[0108] Dosage regimens may be adjusted to provide the optimal desired response. The dosage unit form used herein refers to physically discrete units suited as single dosages for the patients / subjects to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier.

[0109] It should be noted that dosage values ​​may vary depending on the type and severity of the condition to be alleviated and may include single or multiple doses. For any particular subject, a specific dosage regimen should be adjusted over time according to the individual's needs and the professional judgment of the person administering or supervising the administration of the composition. It should be further understood that the dosage ranges described herein are exemplary and are not intended to limit the scope or practice of the compositions embodied. Furthermore, dosage regimens using the compositions of the present application may be based on various variables, including the type of disease, age, weight, sex, the patient's medical condition, the severity of the condition, the route of administration, and the specific antibody used. Therefore, dosage regimens may vary widely but can be routinely determined using standard methods. For example, dosages may be adjusted based on pharmacokinetic or pharmacodynamic parameters, which may include clinical effects such as toxicity and / or experimental values.

[0110] Suitable dosages of the compounds of the present invention are considered to be 0.001-200 mg / kg per day, preferably about 0.01 mg / kg to about 20 mg / kg per day, such as about 0.5-50 mg / kg, for example about 1-20 mg / kg. The compounds may be administered, for example, at least 0.25 mg / kg, such as at least 0.5 mg / kg, such as at least 1 mg / kg, such as at least 1.5 mg / kg, such as at least 2 mg / kg, such as at least 3 mg / kg, such as at least 4 mg / kg, such as at least 5 mg / kg; and, for example, at most 50 mg / kg, such as at most 30 mg / kg, such as at most 20 mg / kg, such as at most 15 mg / kg. Dosages are usually repeated at appropriate intervals, for example, twice daily, three times daily, once daily, once weekly, once every two weeks, or once every three weeks, for as long as the attending physician deems appropriate, and the attending physician may optionally increase or decrease the dosage as necessary.

[0111] The compounds of the present invention can be used in particular for the treatment and prevention, i.e. prophylaxis, of tumor growth and metastasis, and in particular for solid tumors of all indications and stages, regardless of prior treatment of tumor growth.

[0112] Some embodiments of these methods may further comprise administering or treating with one or more additional anticancer therapies. In some of these embodiments, the additional anticancer therapies include surgery, radiation therapy, biotherapy, immunotherapy, chemotherapy, or any combination thereof.

[0113] Some embodiments of these methods may further comprise administration or treatment with one or more anticancer agents. In some of these embodiments, the anticancer agents are chemotherapeutic agents, growth inhibitors, antiangiogenic agents, cytotoxic agents, antihormonal agents, prodrugs, or cytokines.

[0114] In a further embodiment of the present application, the compound of formula (I) of the present application can be used to sensitize cells to radiation, i.e., treating cells with the compound of the present application prior to radiation treatment of the cells renders them more susceptible to DNA damage and cell death than cells not treated with any compound of the present application. In one aspect, the cells are treated with one or more compounds of formula (I) of the present application.

[0115] Accordingly, the present application also provides a method of killing a cell, wherein one or more compounds of the present application are administered to the cell in combination with conventional radiation therapy.

[0116] The present application also provides a method of making a cell more susceptible to apoptosis, wherein the cell is treated with one or more compounds of formula (I) of the present application prior to treatment to cause or induce apoptosis. In one aspect, after the cell is treated with one or more compounds of formula (I) of the present application, the cell is treated with at least one compound, at least one method, or a combination thereof to induce DNA damage for the purpose of inhibiting normal cell function or causing the cell to die.

[0117] In another embodiment of the present application, the cell is treated with at least one DNA damaging agent to cause apoptosis, i.e., the cell is treated with one or more compounds of formula (I) of the present application to sensitize the cell to apoptosis, and then the cell is treated with at least one DNA damaging agent to cause the cell to die. DNA damaging agents useful in the present application include, but are not limited to, chemotherapeutic agents (e.g., cisplatin), ionizing radiation (X-rays, ultraviolet radiation), carcinogens, and mutagens.

[0118] In another embodiment, the cell is killed by treating the cell with at least one method that causes or induces DNA damage. Such methods include, but are not limited to, activating a cellular signaling pathway that causes DNA damage when activated, inhibiting a cellular signaling pathway that causes DNA damage when inhibited, and inducing biochemical changes in the cell that cause DNA damage. As a non-limiting example, inhibition of a cellular DNA repair pathway may inhibit the repair of DNA damage and lead to an abnormal accumulation of DNA damage in the cell.

[0119] In one aspect of the present application, the compound of formula (I) of the present application is administered to the cell prior to radiation or other induction of DNA damage in the cell. In another aspect of the present application, the compound of general formula (I) of the present application is administered to the cell concomitantly with radiation or other induction of DNA damage in the cell. In another aspect of the present application, the compound of formula (I) of the present application is administered to the cell immediately after radiation or other induction of DNA damage in the cell begins.

[0120] In another aspect, the cells are in vitro. In another embodiment, the cells are in vivo. The compounds of the present application may be administered as a single agent or in combination with one or more other pharmaceutically active ingredients, provided that the combination does not cause unacceptable side effects.

[0121] The present application also encompasses such pharmaceutical combinations. For example, the compounds of the present application can be combined with: 131 1-chTNT, abarelix, abiraterone, aclarubicin, adalimumab, ado-trastuzumab emtansine, afatinib, aflibercept, aldesleukin, alectinib, alemtuzumab, alendronic acid, alitretinoin, altretamine, amifostine, aminoglutethimide, hexyl aminolevulinate, amrubicin, amsacrine, Anastrozole, ancestim, anethole dithiolethione, anetumab ravtansine, angiotensin II, antithrombin III, aprepitant, arcitumomab, arglabin, arsenic trioxide, asparaginase, atezolizumab, axitinib, azacitidine, basiliximab, belotecan, bendamustine, besilesomab, belinostat, bevacizumab, bexarotene Bicalutamide, bisantrene, bleomycin,Blinatumomab, bortezomib, buserelin, bosutinib, brentuximab vedotin, busulfan, cabazitaxel, cabozantinib, calcitonine, calcium folinate, calcium levofolinate, capecitabine, capromab, carbamazepine carboplatin, carboquone, carfilzomib, carmofur, carmustine, catumaxomab, celecoxib, celmoleukin, ceritinib, cetuximab, chlorambucil, chlormadinone, chlormethine, cidofovir, cinacalcet, cisplatin, cladribine, clodronic acid, clofarabine, cobimetinib, copanlisib, crisantaspase, crizotinib, cyclophosphamide, cyproterone, cytarabine, dacarbazine, dactinomycin, daratumumab, darbepoetin alfa Dabrafenib, dasatinib, daunorubicin, decitabine, degarelix,Denileukin diftitox, denosumab, depreotide, deslorelin, dianhydrogalactitol, dexrazoxane, dibrospidium chloride, dianhydrogalactitol, diclofenac, dinutuximab, docetaxel, dolasetron, doxifluridine, doxorubicin, doxorubicin + estrone, dronabinol, eculizumab, edrecolomab, elliptinium acetate acetate), elotuzumab, eltrombopag, endostatin, enocitabine, enzalutamide, epirubicin, epitiostanol, epoetin alfa, epoetin beta, epoetin zeta, eptaplatin, eribulin, erlotinib, esomeprazole, estradiol, estramustine, ethinylestradiol, etoposide, everolimus, exemestane, Fadrozole, fentanyl, filgrastim, fluoxymesterone, floxuridine, fludarabine,Fluorouracil, flutamide, folinic acid, formestane, fosaprepitant, fotemustine, fulvestrant, gadobutrol, gadoteric acid meglumine, gadoversetamide, gadoxetic acid, gallium nitrate, ganirelix, gefitinib, gemcitabine, gemtuzumab, glucarpidase, glutoxim, GM-CSF, goserelin, granisetron, Granulocyte colony stimulating factor, histamine dihydrochloride, histrelin, hydroxycarbamide, I-125 seeds, lansoprazole, ibandronic acid, ibritumomab tiuxetan, ibrutinib, idarubicin, ifosfamide, imatinib, imiquimod, improsulfan, indisetron, incadronic acid, ingenol mebutate, interferon alfa, interferon beta, interferon gamma gamma), iobitridol, iobenguane (1231),Iomeprol, ipilimumab, irinotecan, itraconazole, ixabepilone, ixazomib, lanreotide, lansoprazole, lapatinib, lasocholine, lenalidomide, lenvatinib, lenograstim, lentinan, letrozole, leuprorelin, levamisole, levonorgestrel, levothyroxine sodium, lisuride, lobaplatin, lomustine, Lonidamine, masoprocol, medroxyprogesterone, megestrol, melarsoprol, melphalan, mepitiostane, mercaptopurine, mesna, methadone, methotrexate, methoxsalen, methylaminolevulinate, methylprednisolone, methyltestosterone, metyrosine, mifamurtide, miltefosine, miriplatin, mitobronitol, mitoguazone, mitolactol, Mitomycin, mitotane, mitoxantrone, mogamulizumab,molgramostim, mopidamol, morphine hydrochloride, morphine sulfate, nabilone, nabiximols, nafarelin, naloxone + pentazocine, naltrexone, nartograstim, necitumumab, nedaplatin, nelarabine, neridronic acid, netupitant / palonosetron, nivolumab, pentetreotide, nilotinib, nilutamide, nimorazole, nimotuzumab, nimustine, nintedanib, nitracrine, nivolumab, obinutuzumab, octreotide, ofatumumab, olaparib, olaratumab, omacetaxine mepesuccinate, omeprazole, ondansetron, oprelvekin, orgotein, orilotimod, osimertinib, oxaliplatin, oxycodone, oxymetholone, ozogamicine, p53 gene therapy paclitaxel, palbociclib, palifermin, palladium-103 seed,Palonosetron, pamidronic acid, panitumumab, panobinostat, pantoprazole, pazopanib, pegaspargase, PEG-epoetin beta (methoxy PEG-epoetin beta), pembrolizumab, pegfilgrastim, peginterferon alfa-2b, pembrolizumab, pemetrexed, pentazocine, pentostatin, peplomycin, perflubutane, Perfosfamide, Pertuzumab, picibanil, pilocarpine, pirarubicin, pixantrone, plerixafor, plicamycin, poliglusam, polyestradiol phosphate, polyvinylpyrrolidone + sodium hyaluronate, polysaccharide-K, pomalidomide, ponatinib, porfimer sodium, pralatrexate, prednimustine, prednisone, procarbazine, Procodazole, propranolol, quinagolide, rabeprazole, racotumomab,Radium-223 chloride, radotinib, raloxifene, raltitrexed, ramosetron, ramucirumab, ranimustine, rasburicase, razoxane, refametinib, regorafenib, risedronic acid, rhenium-186 etidronate, rituximab, rolapitant, romidepsin, romiplostim, romurtide, roniciclib, samarium (153Sm) samarium (153Sm) lexidronam, sargramostim, satumomab, secretin, siltuximab, sipuleucel-T, sizofiran, sobuzoxane, sodium glycididazole, sonidegib, sorafenib, stanozolol, streptozocin, sunitinib, talaporfin, talimogene laherparepvec, tamibarotene, tamoxifen, tapentadol, tasonermin, Teceleukin, technetium (99mTc) nofetumomab merpentan, 99mTc-HYNIC-[Tyr3]-octreotide, tegafur,Tegafur + gimeracil + oteracil, temoporfin, temozolomide, temsirolimus, teniposide, testosterone, tetrofosmin, thalidomide, thiotepa, thymalfasin, thyrotropin alfa, thioguanine, tocilizumab, topotecan, toremifene, tositumomab, trabectedin, trametinib, tramadol, trastuzumab, trastuzumab emtansine, treosulfan, tretinoin, trifluridine + tipiracil, trilostane, triptorelin, trametinib, trofosfamide, thrombopoietin, tryptophan, ubenimex, valatinib, valrubicin, vandetanib, vapreotide, vemurafenib, vinblastine, vincristine, vindesine, vinflunine, vinorelbine, vismodegib vorinostat, vorozole, yttrium-90 glass microspheres, zinostatin,Zinostatin stimalamer, zoledronic acid, zorubicin.

[0122] The compounds of the present invention may be further combined with other drugs that target the immune system, such as immune checkpoint inhibitors, for example, aPD-1 / -L1 axis antagonists.

[0123] PD-1, along with its ligands PD-L1 and PD-L2, functions as a negative regulator of T cell activation. AHR suppresses immune cell function while increasing cancer cell proliferation and motility. PD-L1 is overexpressed in many cancers, and PD-1 overexpression often occurs concurrently in tumor-infiltrating T cells. Consequently, T cell activation is weakened, immune surveillance is evaded, and anti-tumor immune responses are impaired.

[0124] Simultaneous targeting of the PD-1 / -L1 axis and AHR enhances anti-tumor immune responses beyond additive, leading to unexpected tumor growth reduction.

[0125] Therefore, a composition comprising a PD-1 / -L1 axis antagonist and an AHR antagonist is surprisingly effective in enhancing immune responses and treating cancer.

[0126] Additionally, the compounds of the present application may also be used as therapeutic agents for various other disorders involving AHR.

[0127] Examples of other disorders associated with abnormal AhR signaling inflammation may be one or more selected from the group consisting of infections and cancer vaccinations, viral infections, obesity and diet-induced obesity, adiposity, metabolic disorders, hepatic steatosis and uterine fibroids (uterine leiomyomas or uterine fibroids) in women, chronic kidney disease, acute and chronic renal failure, diabetes, inflammatory or hypertensive kidney disease, heart failure, angina pectoris, hypertension, pulmonary hypertension, ischemia, vascular disease, thromboembolism, arteriosclerosis, sickle cell anemia, erectile dysfunction, benign prostatic hyperplasia, urinary retention disorders associated with benign prostatic hyperplasia, Huntington's, dementia, Alzheimer's and Creutzfeldt-Jakob.

[0128] Also, in another aspect, a pharmaceutical composition is provided comprising an AhR modulator, such as an AhR antagonist of formula (I), and a pharmaceutically acceptable excipient.

[0129] In some aspects, pharmaceutical compositions comprising an AhR modulator, such as an AhR antagonist of formula (I), are provided for use in modulating constitutive AhR activity in a subject in need thereof.

[0130] In some aspects, pharmaceutical compositions comprising an AhR modulator, such as an AhR antagonist of formula (I), are provided for use in treating cancer or a cancer condition by modulating AhR activity.

[0131] In some aspects, pharmaceutical compositions comprising an AhR modulator, e.g., an AhR antagonist of formula (I), are provided for use in inhibiting cancer cell proliferation, tissue invasion, metastasis, and angiogenesis in a subject having cancer, a cancer condition, or a tumor.

[0132]

[0133] aryl hydrocarbon receptor

[0134] The aryl hydrocarbon receptor ("AhR") is a ligand-dependent member of the family of basic-helix-loop-helix transcription factors that has been shown to be activated by a variety of structurally diverse synthetic and naturally occurring compounds, such as polycyclic aromatic hydrocarbons, indoles, and flavonoids. In the absence of bound ligand, AhR resides in a latent state in the cytoplasmic compartment of cells, where it associates with two molecular chaperones, heat shock protein 90 ("hsp90"), an immunophilin-like protein, XAP2, and the hsp90-interacting protein, p23.

[0135] The term "aryl hydrocarbon receptor" or "AhR," as used herein, refers to an 848 amino acid polypeptide, as described, for example, in NP_001612, together with any naturally occurring alleles, splice variants, and engineered forms thereof. Generally, AhR refers to human AhR. The term AhR is also used to refer to truncated forms or fragments of an AhR polypeptide, for example, comprising a specific AhR domain. References to such forms of AhR may be identified herein as "AhR (122-224)".

[0136]

[0137] AhR modulators

[0138] AhR mediates a variety of functional responses, including but not limited to, de novo transcription of AhR battery genes or target genes bearing the DRE or XRE responsive element 5'-TNGCGTG-3'. Alternative pathways of AhR signaling have also been described, such as binding to the retinoblastoma protein, estrogen receptor (ER), transcription factor E2F1, and NFκB pathway subunits RelA and RelB. AhR can also act as a ubiquitin ligase. Thus, signaling through AhR encompasses multiple pathways, including constitutive and non-constitutive AhR signaling pathways or signaling activities, as such terms are defined herein.

[0139] As used herein, “constitutive AhR signaling” refers to one or more signaling pathways that are mediated or regulated by AhR, activated or driven by one or more endogenous AhR ligands, or one or more environmental ligands such as toxins or pollutants, and that result in constitutive or long-term translocation of AhR to the nucleus, and activation or regulation of one or more AhR battery genes involved in unregulated cell growth and proliferation, tumor cell invasiveness, or a combination thereof.

[0140] As used herein, "non-constitutive AhR signaling" refers to one or more signaling pathways mediated or induced by AhR that do not result in constitutive or long-term translocation of AhR to the nucleus, and do not result in activation or regulation of one or more AhR battery genes involved in unregulated cell growth, tumor cell invasiveness, or a combination thereof. In some embodiments, non-constitutive AhR signaling does not result in upregulation of CYP1A1, CYP1B1, or a combination thereof gene expression.

[0141] Accordingly, the term "AhR modulator" as used herein refers to an agent, e.g., a compound of formula (I), that modulates, induces, or promotes a qualitative or quantitative change, alteration, or modification in one or more processes, mechanisms, effects, responses, functions, activities, or pathways mediated by the AhR receptor. Such changes mediated by AhR modulators, such as the AhR antagonists described herein, may mean a decrease, inhibition, or conversion of the constitutive activity of the AhR. The term "expression" refers to the cellular processes involved in producing RNA and proteins, including, but not limited to, secreting proteins, and, where appropriate, transcription, translation, folding, modification, and processing, as applicable. "Expression products" include RNA transcribed from a gene and polypeptides obtained by translation of mRNA transcribed from a gene.

[0142] The term "modulate" in relation to AhR modulators is used consistently with its usage in the art to mean, for example, causing or enabling a qualitative or quantitative change, alteration, or modification in one or more biological processes, mechanisms, effects, responses, functions, activities, pathways, or other phenomena of interest. Thus, as used herein, modulation refers to a qualitative or quantitative change, alteration, or modification in one or more processes, mechanisms, effects, responses, functions, activities, or pathways mediated by the AhR receptor.

[0143] The term "agent" as used herein in connection with an AhR modulator means any compound or substance, such as, but not limited to, a small molecule compound, a nucleic acid, a polypeptide, a peptide, a drug, an ion, and the like. An "agent" can be any chemical, entity, or moiety, and includes, without limitation, synthetic and naturally occurring proteinaceous and non-proteinaceous materials. In some embodiments, the agent is a nucleic acid, a nucleic acid analog, a protein, an antibody, a peptide, an aptamer, an oligomer of a nucleic acid, an amino acid, or a carbohydrate, and includes, without limitation, proteins, oligonucleotides, ribozymes, DNAzymes, glycoproteins, siRNAs, lipoproteins, aptamers, and modifications and combinations thereof. In certain embodiments, as described herein, the agent is a small molecule having a chemical moiety. For example, the chemical moiety comprises an unsubstituted or substituted alkyl, aromatic, or heterocyclyl moiety. Compounds may be known to have the desired activity and / or property (e.g., modulating AhR activity) or may be selected from a library of diverse compounds, for example, using the screening methods described herein.

[0144] In some embodiments, an AhR modulator selectively binds to AhR. As used herein, "selectively binds" or "specifically binds" refers to the ability of an AhR antagonist to bind to a target, such as AhR. D　 10 -5　 M (10000 nM) or less, for example, 10 -6　 M and below, 10 -7　 M and below, 10 -8　 M and below, 10 -9　 M and below, 10 -10　 M and below, 10 -11　 M or less, or 10 -12　 It binds to less than M. For example, the antagonist described herein is 10 -5K below M D An agent binds specifically to AhR if it binds to AhR but not to other molecules or related homologues. Specific binding can be influenced by, for example, the affinity and avidity of the antagonist, and the concentration of the antagonist used. One skilled in the art can determine appropriate conditions under which an antagonist described herein selectively binds using any suitable method, such as those described herein (e.g., titration of an AhR antagonist in an appropriate cell binding assay).

[0145] In some aspects of the compositions and methods described herein, the AhR modulator is an AhR antagonist having the chemical structure of formula (I) described herein.

[0146] As used herein, an AhR modulator is an "AhR antagonist." An AhR antagonist refers to an AhR inhibitor that, when specifically bound to the AhR, does not induce a biological response itself, but blocks or attenuates an agonist- or ligand-mediated response. That is, an AhR antagonist can bind to the AhR but does not activate the AhR; the binding interferes with the interaction, displaces the AhR agonist, and / or inhibits the function of the AhR agonist. Therefore, as used herein, an AhR antagonist does not function as an inducer of AhR activity when bound to the AhR; that is, they function as pure AhR inhibitors. In some embodiments, an AhR antagonist selectively binds to the AhR.

[0147] In some embodiments of these aspects, the AhR antagonists described herein, such as compounds of formula (I), block constitutive AhR effector function that mediates the growth and progression of established tumors. In other embodiments, the small molecule AhR antagonists of formula (I) described herein act as chemopreventive agents by blocking AhR-mediated CYP1A1 induction and mutagenesis upon exposure to environmental ligands.

[0148] In some embodiments of these aspects, the AhR antagonist of Formula (I) described herein inhibits the initial contribution of constitutively active AhR in inducing malignant transformation. In some embodiments, the compound of Formula (I) described herein inhibits constitutive AhR signaling-mediated cancer or tumor cell growth. In some embodiments, the compound of Formula (I) described herein inhibits constitutive AhR signaling-mediated tumor invasion in inducing malignant transformation.

[0149] In one example of the present application, the compound has improved solubility and significantly increased bioavailability, effectively regulating AhR activity, and is therefore useful for preventing or treating various diseases related to AhR activity.

[0150] Hereinafter, the present application will be described in more detail with reference to the following examples. However, these examples are merely intended to illustrate the present application, and the scope of the present application is not limited by these examples.

[0151]

[0152] Manufacturing Example 1. 2-((4-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperazin-1-yl)sulfonyl)ethan-1-ol

[0153]

[0154] Intermediate 1. Nicotinimidamide hydrochloride

[0155] To a suspension of 3-cyanopyridine (5 g, 48.03 mmol) in 50 mL of methanol was added 30 wt% sodium methoxide in methanol (4 mL), and the mixture was stirred at room temperature for 24 hours. After adding ammonium chloride (16.5 g, 0.31 mol), the mixture was heated under reflux for 6 hours and then cooled. The solvent was removed in vacuo, ethanol (60 mL) was added, and the mixture was heated under reflux for 30 minutes. After the reaction mixture was cooled to room temperature, the solid was filtered, and the filtrate was concentrated in vacuo. A suspension of the reaction mixture in 3 mL of ethanol was filtered, and the solid product was dried to obtain 4.9 g of the title compound.

[0156] 1 H NMR (400 MHz, DMSO-d6) δ [ppm] = 9.43 (bs, 4H), 8.98 (d,J= 1.6 Hz, 1H), 8.74 (dd,J= 4.8 Hz,J= 1.2 Hz, 1H), 8.20-8.23 (m, 1H), 7.64-7.67 (m, 1H); MS (ESI, m / z): 122.1 [M+H] +

[0157]

[0158] Intermediate 2. 2-Pyridin-3-yl-pyrimidine-4,6-diol

[0159] To a solution of 3-pyridyl amidine hydrochloride (4.8 g, 30.46 mmol) in methanol (120 mL) was added diethyl malonate (4.63 mL, 30.46 mmol), followed by the addition of a 30 wt% solution of sodium methoxide in methanol (20 mL) at 0°C. The resulting mixture was stirred at room temperature for 24 h. The solvent was removed in vacuo. The resulting residue was used without further purification.

[0160] MS (ESI, m / z): 190.0 [M+H] +

[0161]

[0162] Intermediate 3. 4,6-Dichloro-2-pyridin-3-yl-pyrimidine

[0163] phosphoryl chloride Dimethylaminoaniline (4.77 g, 35.03 mmol) was added to a solution of 2-pyridin-3-yl-pyrimidine-4,6-diol (5.0 g of the crude compound from the previous step) in (10 mL), and the reaction mixture was heated at 120°C for 4 hours. The residue was cooled to room temperature, extracted with 500 mL of ethyl acetate, and concentrated in vacuo. The crude product was purified by silica gel column chromatography to obtain 4.45 g of the title compound.

[0164] 1 H NMR (400 MHz, DMSO-d6) δ [ppm] = 7.36 (s, 1H), 7.49 (dd, J= 4.8 Hz, J= 1.6 Hz, 1H), 8.80-8.72 (m, 2H), 9.64 (br, 1H); MS (ESI, m / z): 226.0 [M+H] +

[0165]

[0166] Intermediate 4. 4-chloro-6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidine

[0167] To a mixture of 4,6-dichloro-2-pyridin-3-yl-pyrimidine (1.03 g, 4.6 mmol), (4-chlorophenyl)boronic acid (0.66 g, 4.2 mmol), and sodium carbonate (1.01 g, 9.5 mmol) in 50 mL of tetrahydrofuran / H2O (4 / 1) was added Pd(PPh3)4 (203 mg, 0.18 mmol). The mixture was heated in a microwave at 80 °C for 20 min, cooled to room temperature, and extracted three times with ethyl acetate (50 mL). The organic layer was dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to obtain 1.02 g of the title compound.

[0168] 1H NMR (600 MHz, CDCl3) δ [ppm] = 7.52 (d, 2H), 7.74 (s, 1H), 7.79 (dd, 1H), 8.06 (d, 2H), 9.02 (d, 1H), 9.12 (d, 1H), 10.16 (s, 1H); MS (ESI, m / z): 302.0 [M+H] +

[0169]

[0170] Manufacturing Example 1. 2-((4-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperazin-1-yl)sulfonyl)ethan-1-ol

[0171] To a solution of 4-chloro-6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidine (45 mg, 0.15 mmol) in tetrahydrofuran (6 mL) was added triethylamine (0.3 mL, 2.15 mmol), followed by 2-(piperazin-1-ylsulfonyl)ethan-1-ol (32 mg, 0.16 mmol) at room temperature. The reaction mixture was heated in a sealed tube at 120 °C for 4 h and cooled to room temperature. The residue was filtered, evaporated in vacuo, and separated by preparative HPLC to give 50 mg of the title compound.

[0172] 1 H NMR (600 MHz, DMSO-d6) δ [ppm] = 9.59 (s, 1H), 8.75 (dd,J= 8.0, 2.0 Hz, 1H), 8.70 (d,J= 4.7 Hz, 1H), 8.37 (d,J= 8.3 Hz, 2H), 7.61 (d,J= 8.3 Hz, 2H), 7.55 (dd,J= 8.0, 4.8 Hz, 1H), 7.45 (s, 1H), 5.04 (s, 1H), 3.97 (s, 4H), 3.75 (t,J= 6.1 Hz, 2H), 3.31 (d,J= 5.1 Hz, 4H), 3.23 (t,J= 6.1 Hz, 2H); MS (ESI, m / z): 460.1 [M+H] +

[0173]

[0174] Manufacturing Example 2. (1-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperidin-4-yl)methanol

[0175]

[0176] To a solution of 4-chloro-6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidine (45 mg, 0.15 mmol) in tetrahydrofuran (6 mL) was added triethylamine (0.3 mL, 2.23 mmol), followed by piperidin-4-ylmethanol (34 mg, 0.30 mmol) at room temperature. The reaction mixture was heated in a sealed tube at 120 °C for 4 h and cooled to room temperature. The residue was filtered, evaporated in vacuo, and purified by preparative HPLC to give 35 mg of the title compound.

[0177] 1 H NMR (400 MHz, CD3OD) δ [ppm] = 1.24-1.34 (m, 2H), 1.82-1.94 (m, 3H), 3.08 (t, 2H), 3.47 (d, 2H), 4.77 (br, 2H), 7.21 (s, 1H), 7.50 (d, 2H), 8.11 (br, 1H), 8.17 (d, 2H), 8.90 (br, 1H), 9.43 (d, 1H), 9.69 (br, 1H); MS (ESI, m / z): 381.1 [M+H] +

[0178]

[0179] Manufacturing Example 3. (R)-1-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)pyrrolidin-3-ol

[0180]

[0181] To a solution of 4-chloro-6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidine (45 mg, 0.15 mmol) in tetrahydrofuran (6 mL) was added triethylamine (0.3 mL, 2.23 mmol), followed by (R)-pyrrolidin-3-ol (19.5 mg, 0.22 mmol) at room temperature. The reaction mixture was heated in a sealed tube at 120°C for 4 h and cooled to room temperature. The residue was filtered, evaporated in vacuo, and purified by preparative HPLC to give 40 mg of the title compound.

[0182] 1 H NMR: (400 MHz, CD3OD) δ [ppm] = 2.17 (br, 2H), 3.70 (br, 2H), 3.86 (br, 2H), 4.59 (br, 1H), 6.95 (s, 1H), 7.52 (d, 2H), 8.07-8.11 (m, 1H), 8.19 (d, 2H), 8.89 (d, 1H), 9.43 (d, 1H), 9.69 (s, 1H); MS (ESI, m / z): 353.1 [M+H] +

[0183]

[0184] Example 1. 2-((4-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperazin-1-yl)sulfonyl)ethyl dihydrogen phosphate

[0185]

[0186] Triethylphosphate (1000 mL, 5885 mmol) was added to the reactor under a nitrogen atmosphere and stirred for more than 10 minutes. While maintaining the internal temperature, 2-((4-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperazin-1-yl)sulfonyl)ethan-1-ol (25 g, 54.35 mmol) obtained in Manufacturing Example 1 was added to the reactor and stirred. After cooling the internal temperature to -6 to 0°C, phosphoryl chloride (15.7 mL, 168.44 mmol) was added dropwise while maintaining the internal temperature. The internal temperature was increased to 0 to 6°C. After stirring for more than 2 hours while maintaining the internal temperature at 0 to 6°C, a process inspection was performed. When the reaction was completed, the internal temperature was cooled to -10 to -4°C. While maintaining the internal temperature at -10 to -4°C, 425 mL of 9% sodium bicarbonate was slowly added dropwise to adjust the pH to 5.0 to 5.5. After increasing the internal temperature to 0 to 10°C, 1125 mL of purified water was added dropwise while maintaining the internal temperature. After cooling the internal temperature to -1 to 5°C, stirring was performed for 2 hours while maintaining the internal temperature. The solid product was filtered, washed with 250 mL of methanol and 250 mL of acetone, dried under reduced pressure at 45 to 55°C for 24 hours, and then left in a silica gel desiccator for 24 hours to obtain 28.7 g (yield 97.8%) of the title compound.

[0187] 1 H NMR (600 MHz, DMSO-d6) δ [ppm] = 9.58 (s, 1H), 8.75 (d,J= 7.9 Hz, 1H), 8.71 (d,J= 4.7 Hz, 1H), 8.38 - 8.33 (m, 2H), 7.62 - 7.58 (m, 2H), 7.58 - 7.53 (m, 1H), 7.42 (s, 1H), 4.15 (q,J= 6.1 Hz, 2H), 3.96 (s, 4H), 3.49 (t,J= 6.1 Hz, 2H), 3.38 - 3.31 (m, 4H); MS (ESI, m / z): 540.1 [M+H] +

[0188]

[0189] Example 2. 2-((4-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperazin-1-yl)sulfonyl)ethyl dihydrogen phosphate dihydrate

[0190]

[0191] The 2-((4-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperazin-1-yl)sulfonyl)ethyl dihydrogen phosphate obtained in Example 1 was ground using a mortar and pestle, and 10.0 g was placed in a constant temperature and humidity chamber. Moisture content test was performed at 12-hour intervals for 12 hours or more at a temperature of 20 to 30°C and a relative humidity of 75±5%. After 24 hours of storage in the constant temperature and humidity chamber, 10.3 g of the title compound (moisture content 6.30%, relative purity 96.92%) was obtained.

[0192]

[0193] Example 3. 2-((4-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperazin-1-yl)sulfonyl)ethyl acetate

[0194]

[0195] 2-((4-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperazin-1-yl)sulfonyl)ethan-1-ol (2 g, 4.35 mmol) and pyridine (60 mL, 744.88 mmol) obtained in Manufacturing Example 1 were added and stirred. Acetyl chloride (1.23 mL, 13.04 mmol) was slowly added at room temperature and stirred for 2 hours. 300 mL of still water was slowly added to the reactor and stirred for 1 hour at room temperature. The solid product was filtered, washed with 100 mL of still water, and dried under reduced pressure at 35 to 45°C for 24 hours to obtain 1.94 g (yield 88.9%) of the title compound.

[0196] 1H NMR (600 MHz, DMSO-d6) δ [ppm] = 9.60 (dd,J= 2.3, 0.9 Hz, 1H), 8.75 (dt,J= 7.9, 1.9 Hz, 1H), 8.71 (dd,J= 4.8, 1.7 Hz, 1H), 8.40 - 8.35 (m, 2H), 7.64 - 7.59 (m, 2H), 7.55 (ddd,J= 7.9, 4.7, 0.9 Hz, 1H), 7.46 (s, 1H), 4.33 (t,J= 6.0 Hz, 2H), 3.99 (s, 4H), 3.49 (t,J= 6.0 Hz, 2H), 3.34 - 3.31 (m, 4H), 1.98 (s, 3H); MS (ESI, m / z): 502.1 [M+H] +

[0197]

[0198] Example 4. 2-((4-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperazin-1-yl)sulfonyl)ethyl methanesulfonate

[0199]

[0200] 2-((4-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperazin-1-yl)sulfonyl)ethan-1-ol (2 g, 4.35 mmol) obtained in Manufacturing Example 1, pyridine (60 mL, 744.88 mmol), and methane sulfonyl chloride (1 mL, 12.92 mmol) were added and stirred at room temperature for 1 hour. 300 mL of constant water was slowly added to the reactor and stirred at room temperature for 1 hour. The solid product was filtered, washed with 100 mL of constant water, and dried under reduced pressure at 35 to 45°C for 24 hours to obtain 2.25 g (yield 96.2%) of the title compound.

[0201] 1H NMR (600 MHz, DMSO-d6) δ [ppm] = 9.60 (dd,J= 2.2, 0.9 Hz, 1H), 8.75 (dt,J= 8.0, 1.9 Hz, 1H), 8.71 (dd,J= 4.8, 1.7 Hz, 1H), 8.40 - 8.34 (m, 2H), 7.64 - 7.59 (m, 2H), 7.55 (ddd,J= 7.9, 4.8, 0.9 Hz, 1H), 7.46 (s, 1H), 4.51 (t,J= 5.9 Hz, 2H), 3.99 (s, 4H), 3.64 (t,J= 5.9 Hz, 2H), 3.38 - 3.35 (m, 4H), 3.24 (s, 3H); MS (ESI, m / z): 538.1 [M+H] +

[0202]

[0203] Example 5. 2-((4-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperazin-1-yl)sulfonyl)ethyl nicotinate

[0204]

[0205] 2-((4-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperazin-1-yl)sulfonyl)ethan-1-ol (1 g, 2.17 mmol) obtained in Preparation Example 1, nicotinic acid (535 mg, 4.35 mmol), 4-dimethylaminopyridine (133 mg, 1.09 mmol), EDC-HCl (1.25 g, 6.52 mmol), and 30 mL of dichloromethane were added to a reactor and stirred overnight at room temperature. After confirming the completion of the reaction, 50 mL of a saturated aqueous sodium bicarbonate solution was added and work-up was performed. The separated organic layer was washed sequentially with 50 mL of water and 50 mL of saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography and then recrystallized using methanol to obtain 1.06 g (yield 86.3%) of the title compound.

[0206] 1H NMR (600 MHz, DMSO-d6) δ [ppm] = 9.59 (d,J= 2.2 Hz, 1H), 9.10 (dd,J= 2.2, 0.9 Hz, 1H), 8.77 (dd,J= 4.8, 1.7 Hz, 1H), 8.74 (dt,J= 7.9, 2.0 Hz, 1H), 8.71 (dd,J= 4.7, 1.7 Hz, 1H), 8.40 - 8.35 (m, 2H), 8.27 (dt,J= 7.9, 2.0 Hz, 1H), 7.64 - 7.60 (m, 2H), 7.55 (ddd,J= 8.0, 4.7, 0.8 Hz, 1H), 7.51 (ddd,J= 7.9, 4.8, 0.9 Hz, 1H), 7.44 (s, 1H), 4.66 (t,J= 5.9 Hz, 2H), 3.99 (s, 4H), 3.67 (t,J= 5.8 Hz, 2H), 3.36 (t,J= 5.1 Hz, 4H); MS (ESI, m / z): 565.2 [M+H] +

[0207]

[0208] Example 6. 2-((4-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperazin-1-yl)sulfonyl)ethyl 4-phenylbutanoate

[0209]

[0210] 2-((4-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperazin-1-yl)sulfonyl)ethan-1-ol (1 g, 2.17 mmol) obtained in Preparation Example 1, 4-phenylbutanoic acid (714 mg, 4.34 mmol), 4-dimethylaminopyridine (133 mg, 1.08 mmol), EDC-HCl (0.834 g, 4.34 mmol), and 30 mL of dichloromethane were added to a reactor and stirred overnight at room temperature. After confirming the completion of the reaction, 50 mL of a saturated aqueous sodium bicarbonate solution was added and workup was performed. The separated organic layer was washed with 50 mL of water and 50 mL of saturated brine in that order, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to obtain 0.61 g (yield 46.0%) of the title compound.

[0211] 1 H NMR (600 MHz, DMSO-d6) δ [ppm] = 9.60 (dd,J= 2.2, 0.9 Hz, 1H), 8.75 (dt,J= 7.9, 2.0 Hz, 1H), 8.71 (dd,J= 4.8, 1.7 Hz, 1H), 8.40 - 8.35 (m, 2H), 7.64 - 7.58 (m, 2H), 7.55 (ddd,J= 7.9, 4.8, 0.9 Hz, 1H), 7.47 (s, 1H), 7.25 - 7.18 (m, 2H), 7.16 - 7.10 (m, 2H), 7.10 - 7.07 (m, 1H), 4.34 (t,J= 5.9 Hz, 2H), 3.99 (s, 4H), 3.49 (t,J= 5.9 Hz, 2H), 3.33 - 3.30 (m, 4H), 2.50 - 2.47 (m, 2H), 2.23 (t,J= 7.4 Hz, 2H), 1.80 - 1.72 (m, 2H); MS (ESI, m / z): 606.14 [M+H] +

[0212]

[0213] Comparative Example 1. (1-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperidin-4-yl)methyl dihydrogen phosphate

[0214]

[0215] Triethylphosphate (145 mL, 855.32 mmol) was added to the reactor under a nitrogen atmosphere and stirred for more than 10 minutes. While maintaining the internal temperature, (1-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperidin-4-yl)methanol (3 g, 7.88 mmol) obtained in Preparation Example 2 was added to the reactor and stirred. After cooling the internal temperature to -6 to 0°C, phosphoryl chloride (2.3 mL, 24.42 mmol) was added dropwise while maintaining the internal temperature. The internal temperature was increased to 0 to 6°C. After stirring for more than 2 hours while maintaining the internal temperature at 0 to 6°C, a process inspection was performed. When the reaction was completed, the internal temperature was cooled to -10 to -4°C. While maintaining the internal temperature at -10 to -4℃, saturated sodium bicarbonate aqueous solution was slowly added dropwise to adjust the pH to 5.0 to 5.5. After increasing the internal temperature to 0 to 10℃, 150 mL of purified water was added dropwise while maintaining the internal temperature. After cooling the internal temperature to -1 to 5℃, stirring was performed for 2 hours while maintaining the internal temperature. The solid product was filtered, washed with 100 mL of purified water and 100 mL of acetone, recrystallized with 60 mL of acetone, and dried under reduced pressure at 30 to 45℃ for 24 hours. 3 g of the obtained crude compound was suspended in 60 mL of DMSO, stirred at an internal temperature of 90℃ for 3 hours, cooled to room temperature, and stirred for an additional hour. 150 mL of a 1 / 3 mixed solvent of water / acetone was slowly added dropwise to the reaction solution, and stirred at room temperature for 1 hour. The produced solid was filtered under reduced pressure, washed sequentially with 60 mL of a 1 / 3 mixed solvent of water / acetone and 60 mL of acetone, and dried under reduced pressure at 35 to 45°C for 24 hours to obtain 2.82 g (yield 77.7%) of the title compound.

[0216] 1 H NMR (600 MHz, DMSO-d6) δ [ppm] = 9.57 (d, J= 2.1 Hz, 1H), 8.77 - 8.67 (m, 2H), 8.39 - 8.31 (m, 2H), 7.62 - 7.49 (m, 3H), 7.37 (s, 1H), 4.74 (s, 2H), 3.73 (t,J= 6.6 Hz, 2H), 3.05 (s, 1H), 2.98 (s, 1H), 1.97 (s, 2H), 1.82 (d,J= 12.9 Hz, 2H), 1.30 - 1.15 (m, 2H); MS (ESI, m / z): 461.1 [M+H] +

[0217]

[0218] Comparative Example 2. (R)-1-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)pyrrolidin-3-yl dihydrogen phosphate

[0219]

[0220] Triethylphosphate (157 mL, 923.25 mmol) was added to the reactor under a nitrogen atmosphere and stirred for more than 10 minutes. While maintaining the internal temperature, (R)-1-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)pyrrolidin-3-ol (3 g, 8.50 mmol) obtained in Preparation Example 3 was added to the reactor and stirred. After cooling the internal temperature to -6 to 0°C, phosphoryl chloride (2.46 mL, 26.36 mmol) was added dropwise while maintaining the internal temperature. The internal temperature was increased to 0 to 6°C. After stirring for more than 2 hours while maintaining the internal temperature at 0 to 6°C, a process inspection was performed. When the reaction was completed, the internal temperature was cooled to -10 to -4°C. While maintaining the internal temperature at -10 to -4℃, saturated sodium bicarbonate aqueous solution was slowly added dropwise to adjust the pH to 5.0 to 5.5. After increasing the internal temperature to 0 to 10℃, 160 mL of purified water was added dropwise while maintaining the internal temperature. After cooling the internal temperature to -1 to 5℃, stirring was performed for 2 hours while maintaining the internal temperature. The solid product was filtered, washed with 100 mL of purified water and 100 mL of acetone, recrystallized with 60 mL of acetone, and dried under reduced pressure at 30 to 45℃ for 24 hours. 3 g of the obtained crude compound was suspended in 60 mL of DMSO, stirred at an internal temperature of 90℃ for 3 hours, cooled to room temperature, and stirred for an additional hour. 150 mL of a 1 / 3 mixed solvent of water / acetone was slowly added dropwise to the reaction solution, and stirred at room temperature for 1 hour. The produced solid was filtered under reduced pressure, washed sequentially with 60 mL of a 1 / 3 mixed solvent of water / acetone and 60 mL of acetone, and dried under reduced pressure at 35 to 45°C for 24 hours to obtain 2.88 g (yield 78.3%) of the title compound.

[0221] 1H NMR (600 MHz, DMSO-d6) δ [ppm] = 9.59 (s, 1H), 8.74 (d,J= 7.9 Hz, 1H), 8.69 (dd,J= 4.7, 1.8 Hz, 1H), 8.34 (d,J= 8.5 Hz, 2H), 7.59 (d,J= 8.5 Hz, 2H), 7.54 (dd,J= 7.9, 4.8 Hz, 1H), 7.06 (s, 1H), 4.98 (d,J= 27.0 Hz, 1H), 4.09 - 3.79 (m, 2H), 3.73 - 3.54 (m, 2H), 2.31 - 2.14 (m, 2H) MS (ESI, m / z) : 433.0 [M+H] +

[0222]

[0223] Test Example 1. Solubility Evaluation

[0224] To evaluate the kinetic solubility, compounds of Preparation Example 1 and Examples 1 and 3 to 6 were dissolved in DMSO to prepare compound solutions at concentrations of 1, 5, and 10 mM, respectively. 198 uL of DW or 0.5 M potassium phosphate buffer (pH 7.4) was added per 2 uL of the prepared compound solutions to obtain final compound concentrations of 10, 50, and 100 uM. These were placed in a V-shaped plate and incubated for 2 hours at 500 rpm and 37°C, and the turbidity at 620 nm was measured using an Envision instrument and compared with that of blank DMSO. If the turbidity was higher than that of blank DMSO, it was judged to be not completely dissolved. The results of the solubility evaluation are shown in Table 1.

[0225] Compound Kinetic Solubility (DW) Kinetic Solubility (pH 7.4) Preparation Example 1 < 10 uM < 10 uMSample 1> 100 uM > 100 uMSample 3 10 - 50 uM < 10 uMSample 4 10 - 50 uM < 10 uMSample 5 10 - 50 uM < 10 uMSample 6 10 - 50 uM < 10 uM

[0226] As shown in Table 1, the compound of Preparation Example 1 had a problem of low solubility, but the compound according to an example of the present application had improved solubility compared to the compound of Preparation Example 1. In particular, the compound of Example 1 had significantly improved solubility and showed the highest solubility, and the compound of Example 2, which is a hydrate form of the compound of Example 1, had higher solubility than the compound of Example 1. Specifically, the compound of Example 1 and the compounds of Examples 3, 4, 5, and 6 have the same parent nucleus but different functional groups bonded to the terminal hydroxyl group, and the compound of Example 1 had a significantly superior solubility improvement effect compared to the compounds of Examples 3, 4, 5, and 6 having the same parent nucleus. Therefore, the compound of Example 1 in which the terminal hydroxyl group was substituted with a dihydrogen phosphate group and the compound of Example 2 in the form of a hydrate thereof had significantly superior solubility improvement effects compared to other substituted compounds.

[0227]

[0228] Test Example 2. Conversion Rate Evaluation

[0229] Human liver microsomes were added to 0.5 mg / mL of 0.1 M KPO4 buffer (pH 7.4) and preincubated at 37°C for 5 minutes. Then, 1% of 0.1 mM DMSO stock of compounds of Examples 1 to 6 and Comparative Examples 1 to 2 was added and incubated at a concentration of 1 uM. 60 minutes after adding the compounds, the reaction was terminated by adding an organic solvent, and the mixture was centrifuged and quantitatively analyzed using LC-MS / MS. At this time, the conversion rate was calculated using the ratio of the concentration of the compound in Preparation Example 1 and the concentration of each compound before the reaction. The results of the conversion rate evaluation are shown in Table 2.

[0230] Compound conversion rate (%) Example 199.6 Example 382.6 Example 40.0 Example 582.3 Example 675.3 Comparative example 162.2 Comparative example 218.0

[0231] As shown in Table 2, it was confirmed that all compounds of Example 1 were converted to compounds of Preparation Example 1 under the experimental conditions. In particular, the compound of Example 1 showed a significantly higher conversion rate compared to compounds of Examples 3, 4, 5, and 6, and considering that Examples 1 to 6 all have the same parent nucleus (compound of Preparation Example 1) structure, the complete conversion of the compound of Example 1 to the compound of Preparation Example 1 is a unique effect. When the conversion rate is high, the bioavailability of the compound of Preparation Example 1 in the body is excellent, and the unpredictable effects on the body caused by the unconverted compound remaining in the body can be minimized. A prodrug is administered into the body in a pharmacologically inactive form and is converted into an active drug through a specific enzyme or chemical reaction, and this conversion process mostly depends on the hydrolysis of a specific chemical bond. Therefore, the hydrate of the prodrug does not affect the recognition and reaction by enzymes, as long as the hydrated state does not significantly change the chemical structure of the drug. Furthermore, if a hydrated prodrug reverts to its original non-hydrated form in the body, the conversion process to the active drug can remain the same. This process is highly dependent on the specificity of the enzyme and the chemical stability of the drug, and thus, the hydrated form of the compound also exhibits a similar conversion rate. Therefore, Compound 2, a hydrated form of Compound 1, can achieve a conversion rate equivalent to that of Compound 1.

[0232] On the other hand, in the case of the compounds of Comparative Examples 1 and 2, the terminal hydroxy of the active drug was substituted with a dihydrogen phosphate group, similar to the compounds of Examples 1 and 2, but complete dephosphorylation was not achieved under the experimental conditions. Therefore, it can be seen that the effect of improving the conversion rate in the body according to phosphorylation of the compound of Preparation Example 1 (Examples 1 and 2) is a specific effect of the compound of Preparation Example 1.

[0233]

[0234] Test Example 3. Bioavailability Evaluation

[0235] The compound of Preparation Example 1 was administered orally once to ICR mice at doses of 100 and 300 mg / kg / 10 mL, and the compound of Example 1 was administered at doses of 117 and 352 mg / kg / 10 mL. At this time, the administered doses were such that the moles of the compound of Preparation Example 1 and the compound of Example 1 were equimolar. Blood was collected through the orbital vein at each designated time, centrifuged to obtain plasma, which was pretreated and analyzed by LC-MS / MS. Pharmacodynamic parameters were calculated using the plasma concentrations. The results of the bioavailability evaluation of the compound of Preparation Example 1 are shown in Table 3.

[0236] Classification Manufacturing Example 1 Example 1 Dose (mg / kg) 100mg / kg 300mg / kg 117mg / kg 352mg / kg Dose (umol / kg) 217umol / kg 652umol / kg 217umol / kg 652umol / kg Terminal t 1 / 2 (h)6.4410.8910.511.1Tmax (h)3.34.03.35.3Cmax (ng / mL)4,0535,29317,96728,100AUC 0-24 (h*ng / mL)42,66764,534245,449419,505AUC 0-last (h*ng / mL)44,47070,159267,697478,168AUC inf (h*ng / mL)46,72685,647319,532593,595

[0237] As shown in Table 3, when the same molar number of the Example 1 compound was administered, a higher exposure in the body was confirmed as the Preparation Example 1 compound compared to when the Preparation Example 1 compound was administered, which can be confirmed from the Cmax and AUC parameters as the Example 1 compound. That is, the higher solubility of the Example 1 compound compared to the Preparation Example 1 compound affected the absorption, and also, since the Tmax of the compound in the body was not delayed compared to when the Preparation Example 1 compound was administered, the Example 1 compound can be quickly converted to the Preparation Example 1 compound in the gastrointestinal tract or body. As a result, when the same molar number of the Example 1 compound was administered orally once, the bioavailability increased compared to when the Preparation Example 1 compound was administered. In addition, the Example 2 compound is in the form of a hydrate of the Example 1 compound, and has a better solubility than the Example 1 compound and a conversion rate at an equivalent level, so the Example 2 compound can achieve a bioavailability at an equivalent level to the Example 1 compound.

[0238]

[0239] Test Example 4. Evaluation of AhR inhibition efficacy

[0240] AhR activation induces target gene expression, such as CYP1A1 and CYP1B1, by binding to AhR-responsive DNA elements, also known as XREs (xenobiotics responsive elements). The assay used to measure AhR activity here is a luciferase assay using cell lines transfected with a luciferase reporter plasmid containing an XRE upstream of a reporter gene. Cells transfected with the XRE-luciferase reporter (XRE-Luc) plasmid drive luciferase activity, which reflects activation and inhibition of AhR in the cells. In addition to transfection with the XRE-reporter vector, cells were co-transfected with a nano-luciferase reporter gene construct (Nano-Luc) containing a constitutively active promoter as an internal control. Kynurenine and kynurenic acid (endogenous AhR agonists) were used to stimulate cells to test the antagonistic properties of the compounds. The half-maximal inhibitory concentration (IC50) and half-maximal effective concentration (EC50) values ​​were calculated using nonlinear regression (four parameters) with Prism8.0 software (GraphPad).

[0241]

[0242] (1) In vitro assay 1: Antagonism in human cell lines

[0243] HepG2 (human hepatoma cell line) cells (Invivogen) stably harboring XRE-luciferase reporter were plated in complete medium and cultured at 37°C in a CO2 incubator. After 24 h, cells were treated with kynurenine (200 μM) alone (negative control) or together with test compounds for 6 h. Luciferase activity was measured using commercial kits such as Promega Luciferase kit or Invivogen Luciferase kit. IC50 values ​​were calculated using luciferase activities. Luciferase activities were further normalized to the kynurenine alone group as the maximum control group and the vehicle group as the minimum control group. The AhR antagonistic potency of compounds in Preparation Example 1 is shown in Table 4. (IC 50 The values ​​are grouped into A, B, C and D, where A: IC 50 < 0.01 μM; B: 0.01 < IC 50 < 0.1 μM; C: 0.1 < IC 50 < 1.0 μM; D: IC 50 >1.0μM)

[0244]

[0245] (2) In vitro assay 2: Antagonism in mouse cell lines

[0246] Hepa1c1c7 (murine hepatoma cell line) cells co-transfected with XRE-Luc and Nano-Luc plasmids were plated in complete medium and cultured overnight at 37°C in a CO2 incubator. After incubation, cells were treated with AhR-activating ligands such as kynurenic acid and kynurenine (#) with or without test compounds for 6 h. Firefly luciferase and Nano-luciferase activities were measured using Nano-glo Luciferase kit (Promega) and relative luciferase activities (Firefly / Nano-Luc) were expressed as IC 50The values ​​were calculated. The relative luciferase activities were further normalized to the agent-only group as the maximum control and the vehicle group as the minimum control. The AhR antagonistic potency of compounds of Preparation Example 1 is shown in Table 4. (IC50 values ​​are grouped into A, B, C and D, where A: IC 50 < 0.01 μM; B: 0.01 < IC 50 < 0.1 μM; C: 0.1 < IC 50 < 1.0 μM; D: IC 50 > 1.0μM)

[0247] Example 1 Analysis 1: AhR-Luc human antagonism (IC 50 , nM)A Assay 2: AhR-Luc mouse antagonism (IC 50 , nM)A

[0248] Therefore, the compound according to one example of the present application, which is converted into the compound of Preparation Example 1 in vivo, had an AhR inhibitory effect.

Claims

1. A compound of the following chemical formula (I), an enantiomer, a diastereomer, a racemate, a solvate, a hydrate, or a pharmaceutically acceptable salt thereof: [Chemical formula (I)] In the above chemical formula (I), X is A or -LA, L is C 1-5 It is alkyl, A is -O(P=O)(OH)2, -O(C=O)CH3, -OSO2CH3, -O(C=O)-pyridinyl, or -O(C=O)-C 1-4 Alkyl-phenyl, Y is a halogen.

2. In paragraph 1, The above A is -O(P=O)(OH)2, A compound, its enantiomer, diastereomer, racemate, solvate, hydrate, or pharmaceutically acceptable salt.

3. In paragraph 1, The above L is C 1-3 alkyl, A compound, its enantiomer, diastereomer, racemate, solvate, hydrate, or pharmaceutically acceptable salt.

4. In paragraph 1, Selected from (1) to (6) below, Compound, its enantiomer, diastereomer, racemate, solvate, hydrate, or pharmaceutically acceptable salt: (1) 2-((4-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperazin-1-yl)sulfonyl)ethyl dihydrogen phosphate; (2) 2-((4-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperazin-1-yl)sulfonyl)ethyl dihydrogen phosphate dihydrate; (3) 2-((4-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperazin-1-yl)sulfonyl)ethyl acetate; (4) 2-((4-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperazin-1-yl)sulfonyl)ethyl methanesulfonate; (5) 2-((4-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperazin-1-yl)sulfonyl)ethyl nicotinate; and (6) 2-((4-(6-(4-chlorophenyl)-2-(pyridin-3-yl)pyrimidin-4-yl)piperazin-1-yl)sulfonyl)ethyl 4-phenylbutanoate.

5. In paragraph 1, The above compound has a solubility in water of 10 uM or more, A compound, its enantiomer, diastereomer, racemate, solvate, hydrate, or pharmaceutically acceptable salt.

6. A pharmaceutical composition comprising a compound according to any one of claims 1 to 5, an enantiomer, a diastereomer, a racemate, a solvate, a hydrate, or a pharmaceutically acceptable salt thereof.

7. In paragraph 6, A pharmaceutical composition for use in the prevention or treatment of a disease mediated by an aryl hydrocarbon receptor (AhR).

8. In paragraph 7, The above aryl hydrocarbon receptor (AhR) mediated disease is a disorder associated with abnormal AhR signaling, pharmaceutical composition.

9. In paragraph 7, A pharmaceutical composition, wherein the disease mediated by the above aryl hydrocarbon receptor (AhR) is a disease mediated by activation of the aryl hydrocarbon receptor (AhR).

10. In paragraph 7, A pharmaceutical composition, wherein the disease mediated by the above aryl hydrocarbon receptor (AhR) is cancer, a malignant tumor, a fibrotic disorder, or a condition having dysregulated immune responses.

11. In paragraph 10, A pharmaceutical composition, wherein the cancer is selected from the group consisting of breast cancer, squamous cell carcinoma, respiratory tract cancer, lung cancer, peritoneal cancer, hepatocellular carcinoma, stomach cancer, pancreatic cancer, brain cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, colon cancer, large intestine cancer, uterine cancer, endometrial cancer, salivary gland carcinoma, kidney or renal cancer, prostate cancer, testicular cancer, vulvar cancer, thyroid cancer, head and neck cancer, B-cell lymphoma, chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), hairy cell leukemia, and chronic myeloblastic leukemia.

12. In paragraph 10, A pharmaceutical composition, wherein the fibrotic disorder is selected from the group consisting of hepatic fibrosis, cirrhosis, pulmonary fibrosis, endomyocardial fibrosis, nephropathy, glomerulonephritis, renal interstitial fibrosis, diabetic fibrotic damage, myelofibrosis, scleroderma, localized scleroderma, keloid, hypertrophic scar, nevus, diabetic retinopathy, proliferative vitreoretinopathy, and sarcoidosis.

13. In paragraph 10, A pharmaceutical composition, wherein the condition having the above dysregulated immune response is selected from the group consisting of sepsis, multiple organ failure, inflammatory disorders of the kidney, chronic intestinal inflammation, pancreatitis, peritonitis, inflammatory skin disorders, inflammatory eye disorders, rheumatic diseases, systemic lupus erythematosus and multiple sclerosis.

14. In paragraph 6, The pharmaceutical composition is used to inhibit proliferation, tissue invasion, metastasis and / or angiogenesis of cancer cells in a subject having cancer or a malignant tumor.

15. A composition for modulating AhR activity, comprising a compound of formula (I) according to any one of claims 1 to 5, or an enantiomer, diastereomer, racemate, solvate, hydrate, or pharmaceutically acceptable salt thereof.

16. A composition according to claim 15, wherein the AhR activity regulation is AhR inhibition.

17. In the terminal hydroxyl group of the compound of the following chemical formula (II), -(P=O)(OH)2, -(C=O)CH3, -SO2CH3, -(C=O)-pyridinyl, and -(C=O)-C 1-4 A method for increasing the solubility of a compound of the following formula (II), comprising the step of introducing a functional group selected from the group consisting of alkyl-phenyl: [Chemical formula (II)] In the above chemical formula (II), LL is a single bond or C 1-5 It is alkyl, YY is a halogen.

18. In paragraph 17, The above functional group is -(P=O)(OH)2.

19. In paragraph 17, The above LL is C 1-3 Alkyl, method.

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