Novel nitric oxide-sensitive compounds and pharmaceutical compositions containing the same

Abstract

This invention relates to a compound in which a nitric oxide scavenging agent is bound to a pharmaceutically active low-molecular-weight compound, and to a nitric oxide-sensitive pharmaceutical composition containing the same. The compound is designed so that the nitric oxide scavenging agent is separated while removing nitric oxide only at disease sites such as inflamed areas where the concentration of nitric oxide is high, thereby causing the inactivated low-molecular-weight compound to exhibit pharmaceutically active properties. The compound is activated only at disease sites, minimizing the side effects of existing pharmaceuticals, while simultaneously ensuring the pharmaceutically active effect of the low-molecular-weight compound and the effect of nitric oxide removal, making it usable for the prevention or treatment of diseases involving nitric oxide generation.

Description

[Technical Field]

[0001] This application claims priority based on Korean Patent Application No. 10-2023-0086949, filed with the Korean Intellectual Property Office on July 5, 2023, the contents of which are incorporated into this application by reference herein.

[0002] The present invention relates to a nitric oxide-sensitive pharmaceutical composition, wherein the pharmaceutical composition comprises a nitric oxide scavenging agent site that senses for and removes nitric oxide, and a low-molecular-weight compound having another pharmaceutically active component. In normal tissues where the concentration of nitric oxide is low, the activity of the pharmaceutically active low-molecular-weight compound is minimized. In inflammatory disease sites where the concentration of nitric oxide is high, the nitric oxide scavenging agent is separated from the low-molecular-weight compound while removing nitric oxide. This allows the pharmaceutically active component of the low-molecular-weight compound to be locally and selectively present only at the target site, thereby minimizing side effects and maximizing efficacy. This composition can be usefully used for the prevention or treatment of all diseases related to nitric oxide. [Background technology]

[0003] Nitric oxide is a highly reactive radical molecule with a short half-life of approximately 5 seconds or less, and acts as a major vehicle for cellular signaling, linked to a variety of diseases including inflammatory diseases, cancer growth, and metastasis.

[0004] Specifically, inflammatory diseases associated with nitric oxide include rheumatoid arthritis and osteoarthritis, asthma, inflammatory bowel disease (Ulcerative Colitis) and Crohn's disease, inflammatory diseases caused by bacterial infections, inflammatory diseases caused by viral infections, inflammatory diseases caused by parasitic infections, and septic shock. Neurological diseases such as Alzheimer's disease, amyotrophic sclerosis, Huntington's disease, Parkinson's disease, and stroke have also been linked to nitric oxide. Furthermore, cardiovascular diseases such as hypertension and heart failure, as well as diseases such as cancer, obesity, and hepatic encephalopathy, are also known to be associated with nitric oxide.

Summary of the Invention

Problems to be Solved by the Invention

[0005] An object of the present invention is to provide a pharmaceutical composition for preventing or treating all diseases related to nitric oxide, by minimizing the side effects and toxicity of a low-molecular compound and maximizing the drug efficacy, wherein the nitric oxide-sensitive pharmaceutical composition locally and selectively removes nitric oxide only at a diseased site with a high concentration of nitric oxide and releases a low-molecular compound having pharmaceutical activity.

[0006] However, the problems to be solved by the present invention are not limited to the above-mentioned problems, and other problems not mentioned will be clearly understood by those skilled in the art from the following description.

Means for Solving the Problems

[0007] All combinations of various elements disclosed in the present invention belong to the scope of the present invention. Also, the scope of the present invention is not necessarily limited by the following specific description.

[0008] To achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating all diseases related to nitric oxide, comprising as an active ingredient a low-molecular compound to which a nitric oxide scavenger is bound.

[0009] The compound according to the present invention is a substance obtained by binding a nitric oxide scavenger and a low-molecular compound having pharmaceutical activity, and the amine group or hydroxyl group present in the low-molecular compound can be bound to the nitric oxide scavenger using an organic synthesis reaction.

[0010] One embodiment of the present invention provides a compound represented by the following Chemical Formula 1, or a solvate, hydrate, stereoisomer, or pharmaceutically acceptable salt thereof:

[0011]

Chem.

[0012] In the above chemical formula 1, X is a low-molecular-weight compound having pharmaceutically active properties and containing an amine group or a hydroxyl group in the molecular structure of the compound, and may include, for example, kinase inhibitors, particularly Janus kinase inhibitors (JAK inhibitors), other anticancer agents, anti-inflammatory agents, and the like.

[0013] The kinase inhibitors mentioned above may include netarsudil (ROCK1 / 2, a glaucoma treatment agent), fostamatinib (Syk, a thrombocytopenia treatment agent), and belumosudil (ROCK2, a graft-versus-host disease treatment agent).

[0014] The Janus kinase inhibitor may be selected from the group consisting of tofacitinib, upadacitinib, baricitinib, filgotinib, abrocitinib, delgocitinib, oclacitinib, peficitinib, and ruxolitinib.

[0015] The aforementioned anticancer drugs include doxorubicin, cyclophosphamide, cisplatin, oxaliplatin, 5-Fu (5-Fluorouracil), gemcitabine, paclitaxel, docetaxel, irinotecan, and MMAE (Monomethyl auristatin). E) Crizotinib, Osimertinib, Sorafenib, Ibrutinib, Ruxolitinib, Vemurafenib, Ceritinib, Alectinib, Brigatinib, Lorlatinib ), Capmatinib, Gefitinib, Erlotinib, Lapatinib, Icotinib, Afatinib, Neratinib, Dacomitinib, Almonertinib, Tucatinib, Midostaurine ( Midostaurin), Gilteritinib, Quizartinib, Pexidartinib, Sunitinib, Pazopanib, Vandetanib, Axitinib, Cabozantinib, Regorafenib, A Apatinib, Lenvatinib, Tivozanib, Fruquintinib, Nintedanib, Anlotinib, Erdafitinib, Pemigatinib, Avapritinib, Ripretinib,Pralsetinib, Larotrectinib, Entrectinib, Imatinib, Dasatinib, Nilotinib, Bosutinib, Radotinib, Ponatinib, Acalabrutinib, Zanubrutinib, Fedratinib, Dabrapheni Dabrafenib, Encorafenib, Trametinib, Cobimetinib, Binimetinib, Selumetinib, Palbociclib, Ribociclib, Abemaciclib, Idelalisib, Copanlisib, Duvelisib, Alpelisib pelisib), Tazemetostat, Vorinostat, Belinostat, Tucidinostat, Panobinostat, Enasidenib, Ivosidenib, Venetoclax, Vismodegib, Sonidegib, Glasdegib, Bortezomib ezomib), carfilzomib, ixazomib, olaparib, rucaparib, niraparib, talazoparib, umbralisib, trilaciclib, infigratinib, mobocertinib, asciminib, futibatinib,The drug may be selected from a group consisting of pacritinib and everolimus, among others.

[0016] The aforementioned anti-inflammatory agent may be selected from the group consisting of dexamethasone, methotrexate, cyclosporine, acetaminophen, etodolac, piroxicam, aceclofenac, and others.

[0017] In the above chemical formula 1, Y is hydrogen, a C1-C6 alkyl group, a C1-C6 alkoxy group, a hydroxyl group, a nitro group, an amino group, a halogen, a thiol group, a cyano group, and [ka] You may choose from the following options.

[0018] The aforementioned n is an integer between 0 and 4.

[0019] The aforementioned R1 is a C1-C6 alkyl group, a C1-C6 alkoxy group, a hydroxyl group, a nitro group, an amino group, a halogen, a thiol group, a cyano group, or a C3-C 14 Aryl group, C3-C 14 heteroaryl group, -NR 11 R 12 , -NR 12 C(=O)R 12 -C(=O)R 11 , and -C(=O)OR 11 They may be selected from among the following, and if there are two or more R1s, they may be identical or different from each other, and two adjacent R1s may be optionally bonded to form a fused ring. Here, each aryl group or heteroaryl group independently has at least one substituent selected from the group consisting of halogens, hydroxyl groups, thiol groups, cyano groups, C1-C6 alkyl groups, and C1-C6 alkoxy groups, or is unsubstituted, and R11 and R 12 is each independently selected from hydrogen, halogen, CF3, C1-C6 alkyl group, C3-C 14 aryl group, and C3-C 14 heteroaryl group.

[0020] In Chemical Formula 1, Z is

Chemical Formula

[0021] In Chemical Formula 1, m is an integer from 0 to 4.

[0022] R2 is a C1-C6 alkyl group, a C1-C6 alkoxy group, a hydroxy group, a nitro group, an amino group, a halogen, a thiol group, a cyano group, a C3-C 14 aryl group, a C3-C 14 heteroaryl group, -NR 21 R 22 , -NR 22 C(=O)R 22 , -C(=O)R 21 , and -C(=O)OR 21 and may be selected from among them. When there are two or more R2s, they may be the same or different from each other, and two adjacent R2s may optionally combine to form a fused ring. Here, the aryl group or heteroaryl group each independently has at least one substituent selected from the group consisting of halogen, hydroxy group, thiol group, cyano group, C1-C6 alkyl group, and C1-C6 alkoxy group, or is unsubstituted, and R 21 and R 22 is each independently selected from hydrogen, halogen, CF3, C1-C6 alkyl group, C3-C 14 aryl group, and C3-C 14 heteroaryl group.

[0023] R2 may be the same as the definition of R1. The R 21 and the R 22Each of the above R 11 and R 12 It may be the same as the definition of [the other term].

[0024] R3 and R4 can each be independently selected from hydrogen, halogen, hydroxyl group, thiol group, cyano group, C1-C6 alkyl group, and C1-C6 alkoxy group, or R3 and R4 can bond together to form a fused ring.

[0025] Other embodiments of the present invention provide compounds represented by the following chemical formula 2, or solvates, hydrates, stereoisomers thereof, or pharmaceutically acceptable salts thereof:

[0026] [ka]

[0027] In the above chemical formula 2, R2 and m are the same as defined in the above chemical formula 1.

[0028] The compound represented by chemical formula 2 above may be tofacitinib, or one of the compounds listed in Table 1 below, or a solvate, hydrate, stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0029] [Table 1-1] [Table 1-2]

[0030] The compound represented by chemical formula 2 above may be upadacitinib, or one of the compounds listed in Table 2 below, or a solvate, hydrate, stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0031] [Table 2]

[0032] The compound represented by chemical formula 2 may be dexamethasone, or one of the compounds listed in Table 3 below, or a solvate, hydrate, stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0033] [Table 3]

[0034] The compound represented by chemical formula 2 may be cyclosporine, or one of the compounds listed in Table 4 below, or a solvate, hydrate, stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0035] [Table 4]

[0036] Other embodiments of the present invention provide compounds represented by the following chemical formula 3, or solvates, hydrates, stereoisomers thereof, or pharmaceutically acceptable salts thereof:

[0037] [ka]

[0038] In the above chemical formula 3, R1, R2, n, and m are the same as defined in the above chemical formula 1.

[0039] The compound represented by chemical formula 3 may be tofacitinib, or one of the compounds listed in Table 5 below, or a solvate, hydrate, stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0040] [Table 5]

[0041] Other embodiments of the present invention provide compounds represented by the following chemical formula 4, or solvates, hydrates, stereoisomers thereof, or pharmaceutically acceptable salts thereof:

[0042] [ka]

[0043] In the aforementioned chemical formula 4, R2 and m are the same as defined in the aforementioned chemical formula 1.

[0044] The compound represented by chemical formula 4 may be tofacitinib, or one of the compounds listed in Table 6 below, or a solvate, hydrate, stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0045] [Table 6]

[0046] Other embodiments of the present invention provide compounds represented by the following chemical formula 5, or solvates, hydrates, stereoisomers thereof, or pharmaceutically acceptable salts thereof:

[0047] [ka]

[0048] In the above chemical formula 5, R2, R3, R4, and m are the same as defined in the above chemical formula 1.

[0049] The compound represented by chemical formula 5 may be tofacitinib, or one of the compounds listed in Table 7 below, or a solvate, hydrate, stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0050] [Table 7]

[0051] Other embodiments of the present invention provide compounds represented by the following chemical formula 6, or solvates, hydrates, stereoisomers thereof, or pharmaceutically acceptable salts thereof:

[0052] [ka]

[0053] In the aforementioned chemical formula 6, R2, R3, R4, and m are the same as defined in the aforementioned chemical formula 1.

[0054] The compound represented by chemical formula 6 may be tofacitinib, or one of the compounds listed in Table 8 below, or a solvate, hydrate, stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0055] [Table 8]

[0056] The compound represented by chemical formula 6 may be upadacitinib, or one of the compounds listed in Table 9 below, or a solvate, hydrate, stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0057] [Table 9]

[0058] Another embodiment of the present invention provides a pharmaceutical composition for the prevention or treatment of diseases involving the generation of nitric oxide, comprising a compound represented by chemical formula 1, or a solvate, hydrate, stereoisomer of said compound, or a pharmaceutically acceptable salt thereof. Specifically, the pharmaceutical composition for the prevention or treatment of diseases involving the generation of nitric oxide may contain at least one of the compounds represented by chemical formulas 1 to 6, or a solvate, hydrate, stereoisomer thereof, or a pharmaceutically acceptable salt thereof. The pharmaceutical composition may be a nitric oxide-sensitive pharmaceutical composition. The nitric oxide-sensitive pharmaceutical composition may remove at least a portion of nitric oxide at a disease site involving the generation of nitric oxide, and release a compound having pharmaceutically active properties at the disease site.

[0059] The diseases associated with the generation of nitric oxide may be selected from the group consisting of inflammatory diseases, neurological diseases, cardiovascular diseases, autoimmune diseases, allergic diseases, cancer, obesity, myelofibrosis, and hepatic encephalopathy.

[0060] The aforementioned inflammatory diseases may be selected from the group consisting of all inflammatory diseases caused by high concentrations of nitric oxide, including inflammatory diseases due to infection (inflammatory diseases due to viral infection, inflammatory diseases due to bacterial infection, sepsis, etc.), degenerative inflammatory diseases (osteoarthritis), inflammatory diseases due to injury, autoimmune diseases (rheumatoid arthritis, ulcerative bowel disease, Crohn's disease, systemic lupus erythematosus, psoriasis, multiple sclerosis, idiopathic arthritis in children), allergic diseases (asthma, rhinitis, atopic dermatitis), cancer (lung cancer, breast cancer, colorectal cancer, stomach cancer, liver cancer, brain cancer, pancreatic cancer, thyroid cancer, skin cancer, bone marrow cancer, lymphoma, uterine cancer, cervical cancer, ovarian cancer, kidney cancer, melanoma, etc.), and other diseases caused by abnormal hyperactivation of immune cells such as myelofibrosis.

[0061] The aforementioned neurological disorders may be selected from the group consisting of Alzheimer's disease, amyotrophic sclerosis, Huntington's disease, Parkinson's disease, and stroke.

[0062] The aforementioned cardiovascular disease may be hypertension or heart failure.

[0063] The pharmaceutical composition may be administered orally or parenterally. When the pharmaceutical composition is administered parenterally, it may be administered intra-articularly, intravenously, subcutaneously, intramuscularly, intraperitoneally, intranasally, intrapulmonaryly, or rectally.

[0064] The pharmaceutical composition may be administered orally (e.g., in the form of tablets or capsules), parenterally (e.g., in the form of injectable solutions or suspensions), topically (e.g., in the form of lotions, gels, ointments or creams), or intranasally or in the form of suppositories, by any conventional route. A pharmaceutical composition containing the compound of the present invention in free form or in the form of a pharmaceutically acceptable salt, combined with at least one pharmaceutically acceptable carrier or diluent, may be manufactured by conventional methods such as mixing, granulation, or coating. For example, oral compositions may be tablets, granules, or capsules containing excipients, disintegrants, binders, lubricants, etc., and the active ingredient. Injectable compositions may be solutions or suspensions, which may be sterile and may contain preservatives, stabilizers, buffers, etc.

[0065] To obtain the desired therapeutic effect by using the pharmaceutical composition in actual treatment, the dosage of the active ingredient, the compound represented by Chemical Formula 1 of the present invention, or its solvates, hydrates, stereoisomers, or pharmaceutically acceptable salts thereof, can be determined according to the patient's age, sex, weight, the type of disease involving the generation of nitric oxide, and the degree of treatment. For example, in the case of oral administration, for an adult (assuming a weight of 60 kg), the dosage can be in the range of approximately 0.001 to 3,000 mg / kg per day, administered once or in several divided doses, and can be administered orally or parenterally once or multiple times every two days, per week, or per month. It will be understood that the dosage can be determined according to the type of low molecular weight compound represented by X. The dosage for a particular individual or patient must be determined in light of various relevant factors such as the patient's weight, age, sex, health status, prescribed diet, administration time, method of administration, and severity of the disease, and it should be understood that it can be appropriately adjusted by a professional, and the dosage is not intended to limit the scope of the present invention in any way.

[0066] The compounds of the present invention can be used in combination with other therapeutic agents. These compounds can be administered together with therapeutic agents selected from the group consisting of cytotoxic drugs, radiotherapy, and immunotherapy.

[0067] Another embodiment of the present invention provides a method for treating a disease involving the generation of nitric oxide, comprising the step of administering a pharmaceutical composition containing the compound represented by chemical formula 1, or a solvate, hydrate, stereoisomer of the compound, or a pharmaceutically acceptable salt thereof, to a subject. [Effects of the Invention]

[0068] The pharmaceutical composition for the prevention or treatment of diseases involving the generation of nitric oxide according to the present invention can be used as a preventive or therapeutic agent for diseases by selectively releasing the drug only to disease sites where the concentration of nitric oxide is high in diseases related to nitric oxide. [Brief explanation of the drawing]

[0069] [Figure 1] Figure 1 is a conceptual diagram illustrating the operating principle of a nitric oxide-sensitive pharmaceutical composition according to one embodiment of the present invention.

[0070] [Figure 2A] Figure 2 shows the HPLC results of compound (4) prepared in Example 4, with and without nitric oxide. Figure 2A shows the HPLC results of compound (4) in an environment without nitric oxide. [Figure 2B] Figure 2B shows the HPLC results of the reaction of compound (4) in an environment with excess nitric oxide. [Figure 2C] Figure 2C shows the HPLC results for tofacitinib.

[0071] [Figure 3] Figure 3 shows the NMR data of compound (35) prepared in Example 35.

[0072] [Figure 4] Figure 4 is a graph showing the efficacy evaluation of compound (1) produced in Example 1 for inflammatory bowel disease.

[0073] [Figure 5] Figure 5 is a graph showing the efficacy evaluation of compound (4) produced in Example 4 for inflammatory bowel disease.

[0074] [Figure 6] Figure 6 is a graph showing the efficacy evaluation of compound (1) produced in Example 1 against rheumatoid arthritis.

[0075] [Figure 7] Figure 7 is a graph showing the efficacy evaluation of compound (11) produced in Example 11 against rheumatoid arthritis. [Modes for carrying out the invention]

[0076] The present invention can be more readily understood by referring to the following detailed descriptions of preferred embodiments and examples of the invention included in this application. It should be understood that the terms used in this application are for the purpose of describing specific embodiments only and are not intended to be limiting. Furthermore, unless specifically defined in this application, the terms used in this application should be understood to provide the traditional meaning known in the relevant art.

[0077] The singular form used in this application includes plural forms unless otherwise indicated. For example, substituents include one or more substituents.

[0078] In this specification, the term "halo" or "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br), or iodine (I), unless otherwise specified.

[0079] In this specification, the term “alkyl” refers to saturated monovalent aliphatic hydrocarbon radicals, including linear and branched chains having a specific number of carbon atoms, unless otherwise specified. Alkyl alkyl groups typically have 1 to 20 carbon atoms ("C1-C20"). 20 Alkyl), preferably 1 to 12 carbon atoms ("C1-C") 12 Alkyl groups include, more preferably, 1 to 8 carbon atoms ("C1-C8 alkyl"), 1 to 6 carbon atoms ("C1-C6 alkyl"), or 1 to 4 carbon atoms ("C1-C4 alkyl"). Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, n-heptyl, n-octyl, and the like.

[0080] In this specification, the term “alkoxy” refers to a monovalent -O-alkyl group having a specific number of carbon atoms in the alkyl moiety, unless otherwise specified. Alkoxy groups typically have 1 to 8 carbon atoms ("C1-C8 alkoxy"), 1 to 6 carbon atoms ("C1-C6 alkoxy"), or 1 to 4 carbon atoms ("C1-C4 alkoxy"). For example, C1-C4 alkoxys include methoxy (-OCH3), ethoxy (-OCH2CH3), isopropoxy (-OCH(CH3)2), and tert-butyloxy (-OC(CH3)3). The alkoxy groups are suitable for alkyl groups and may be substituted or unsubstituted on the alkyl moiety with the same groups described herein. In particular, the alkoxy groups may be optionally substituted with one or more halo atoms, especially one or more fluoro atoms, up to the total number of hydrogen atoms present on the alkyl moiety. Such groups are called "haloalkoxy" groups, which have a specific number of carbon atoms and are substituted with one or more halo substituents. For example, when fluorinated, they are more specifically called "fluoroalkoxy" groups. Typically, such groups contain 1 to 6 carbon atoms, preferably 1 to 4 carbon atoms, often 1 or 2 carbon atoms, and 1, 2 or 3 halo atoms (i.e., "C1-C6 haloalkoxy", "C1-C4 haloalkoxy", or "C1-C2 haloalkoxy"). More specifically, fluorinated alkyl groups may be specifically called fluoroalkoxy groups, which are typically substituted with 1, 2 or 3 fluoro atoms, for example, C1-C6, C1-C4, or C1-C2 fluoroalkoxy groups. Thus, C1-C4 fluoroalkoxy groups include trifluoromethyloxy (-OCF3), difluoromethyloxy (-OCF2H), fluoromethyloxy (-OCFH2), difluoroethyloxy (-OCH2CF2H), and the like.

[0081] In this specification, the terms “aryl” or “aromatic” refer to an optionally substituted or unsubstituted monocyclic, fused dicyclic, or polycyclic ring system having well-known aromatic characteristics, where one or more rings comprise a fully conjugated pi-electron system. Typically, an aryl group has 3 to 20 carbon atoms as ring members ("C3-C3").20 "aryl"), preferably 6 to 14 carbon atoms ("C6-C") 14 "aryl"), more preferably 6 to 12 carbon atoms ("C6-C") 12 The aryl group has an aryl group. The fused aryl group may be fused to another aryl or heteroaryl ring, or to a saturated or partially unsaturated carboncyclic or heterocyclic ring (e.g., a phenyl ring), and the attachment sites on such a fused ring system to the base molecule are atoms of the aromatic portion of the ring system. Examples of aryl groups include phenyl, biphenyl, naphthyl, toluyl, anthracenyl, phenantrenyl, indanyl, indenyl, or tetrahydronaphthyl. The aryl group may be substituted or unsubstituted, as further described herein.

[0082] In this specification, the terms “heteroaryl” or “heteroaromatic” refer to monocyclic, fused dicyclic, or polycyclic ring systems having a well-known aromatic characteristic, comprising a specific number of ring atoms as ring members in an aromatic ring, and containing one or more heteroatoms selected from B, N, O, S, P, and Si. The inclusion of heteroatoms allows for aromaticity in 5-membered and 6-membered rings. Typically, heteroaryl groups have 5 to 20 ring atoms ("5-20 membered heteroaryl"), preferably 5 to 14 ring atoms ("5-14 membered heteroaryl"), and more preferably 5 to 12 membered ring atoms ("5-12 membered heteroaryl"). Heteroaryl rings attach to the base molecule via the ring atoms of the heteroaromatic ring in such a way that aromaticity is maintained. Thus, a 6-membered heteroaryl ring can attach to the base molecule via the ring C atom, while a 5-membered heteroaryl ring can attach to the base molecule via the ring C or N atom. Heteroaryl groups can also fuse to other aryl or heteroaryl rings, or to saturated or partially unsaturated carboncyclic or heterocyclic rings, and the attachment sites to the underlying molecule on such fused ring systems are atoms of the heteroaromatic moiety of the ring system. For example, unsubstituted heteroaryl groups include pyrrole, furan, thiophene, pyrazole, imidazole, isoxazole, oxazole, isothiazole, thiazole, triazole, oxadiazole, thiadiazole, tetrazole, pyridine, pyridazine, pyrimidine, pyrilazine, benzofuran, benzothiophene, indole, benzimidazole, indazole, quinoline, isoquinoline, purine, triazine, naphthyridine, or carbazole. In various embodiments, the 5- or 6-membered heteroaryl group is selected from the group consisting of pyrrolyl, furanyl, thiophenyl, pyrazolyl, imidazolyl, isoxazolyl, oxazolyl, isothiazolyl, thiazolyl, triazolyl, pyridinyl, and pyrimidinyl, pyrazinyl, or pyridazinyl rings. The heteroaryl group is substituted or unsubstituted, as further described herein.

[0083] The present invention will be described in detail below with reference to examples and experimental examples. However, the following examples and experimental examples are merely illustrative of the present invention and do not limit the scope of the present invention.

[0084] Example 1: Preparation of compound (1)(N-(2-aminophenyl)-4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide)

[0085] [ka]

[0086] Step (1): Production of compound (1-1)

[0087] 300 mg of tofacitinib (0.96 mmol, 1 equiv) and 394 mg of bis(4-nitrophenyl)carbonate (1.30 mmol, 1.35 equiv) were added to 8 mL of acetonitrile, and the mixture was cooled to 0-5°C. 229 μL of DIPEA (1.34 mmol, 1.4 equiv) was added to the mixture, and the mixture was stirred at room temperature for 4 hours. After the mixture was cooled to 0-5°C, 214 mg of tert-butyl(2-aminophenyl)carbamate (0.96 mmol, 1 equiv) was dissolved in 6 mL of dichloromethane and added dropwise, and the mixture was stirred at room temperature for 16 hours. After the reaction was complete, water and dichloromethane were added to the reaction mixture for extraction. The organic layer was separated, dried over magnesium sulfate, filtered, and the remaining filtrate was concentrated under reduced pressure. The concentrated solution was purified by silica gel column chromatography (ethyl acetate:hexane = 1:1) to obtain compound (1-1) of the target compound. (Yield 19%)

[0088] Step (2): Production of compound (1)

[0089] 34 mg of compound (1-1) (0.06 mmol, 1 equiv) was dissolved in 1 mL of dioxane, and the mixture was cooled to 0-5°C. 130 μL of hydrochloric acid (4 M dioxane solution, 0.44 mmol, 7.25 equiv) was added to the mixture, and the mixture was stirred at room temperature for 16 hours. After the reaction was complete, the reaction solution was cooled to 0-5°C, and neutralized by adding 5% NaHCO3 aqueous solution dropwise. Water and ethyl acetate were added to the mixture and extracted. After separating the organic layer, it was dried over magnesium sulfate, filtered, and the remaining filtrate was concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (ethyl acetate:hexane = 1:1) to obtain the target compound (1). (Yield 99%)

[0090] 1 H-NMR(300MHz,DMSO-d6)δ11.55(s,1H),8.42(d,1H,J=3Hz),8.12(d,1H,J=9Hz),7.77(d,1H,J=3Hz),7.61(d,1H,J=6Hz),7.29-7.1 8(m,3H),6.96-6.91(m,2H),4.21-3.62(m,4H),3.49-3.38(m,2H),3.33(s,3H),2.41(m,1H),1.98-1.60(m,3H),1.02(d,3H,J=6Hz).

[0091] Example 2: Preparation of compound (2)(N-(2-amino-5-methylphenyl)-4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide)

[0092] [ka]

[0093] Step (1): Production of tert-Butyl(4-methyl-2-nitrophenyl)carbamate

[0094] 1 g of 4-methyl-2-nitroaniline (6.57 mmol, 1 equiv) was dissolved in 18 mL of anhydrous THF, and the mixture was cooled to 0-5°C. 578 mg of NaH (60% content) (14.46 mmol, 2.2 equiv) was added, and the mixture was stirred for 10 minutes. The mixture was heated to room temperature and stirred for 30 minutes. 1.58 g of (Boc)2O (7.23 mmol, 1.1 equiv) dissolved in 1.8 mL of anhydrous THF was added to the mixture, and the mixture was stirred for 16 hours. After the reaction was complete, water and ethyl acetate were added to the reaction mixture and extracted. The organic layer was separated, dried over magnesium sulfate, filtered, and the remaining filtrate was concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (ethyl acetate:hexane = 1:100) to obtain the target compound, tert-butyl(4-methyl-2-nitrophenyl)carbamate. (Yield 92%)

[0095] Step (2): Production of compound (2-1)

[0096] 465 mg of tert-butyl(4-methyl-2-nitrophenyl)carbamate (1.84 mmol, 1 equiv) was dissolved in 32 mL of methanol. 47 mg of 10% Pd / C was added to the mixture at room temperature, and the mixture was stirred under atmospheric pressure and hydrogen gas for 2 hours. After the reaction, the mixture was purified using a Celite filter and washed with ethyl acetate. The filtrate was concentrated under reduced pressure to obtain compound (2-1), the target compound. (Yield 99%)

[0097] Step (3): Production of compound (2-2)

[0098] In step (1) of Example 1, compound (2-1) was used instead of tert-butyl(2-aminophenyl)carbamate, and the same procedure was repeated to obtain compound (2-2), the target compound. (Yield 74%)

[0099] Step (4): Production of compound (2)

[0100] In step (2) of Example 1, compound (2-2) was used instead of compound (1-1), and the same procedure was repeated to obtain compound (2), the target compound. (Yield 42%)

[0101] 1 H-NMR(300MHz,DMSO-d6)δ11.21(s,1H),8.38(d,1H,J=3Hz),7.74(d,1H,J=3Hz),7.29(s,1H),6.91-6.72(m,3H) ,4.79(s,2H),4.13-3.42(m,6H),3.32(s,3H),2.49(m,1H),2.19(s,3H),1.89-1.59(m,3H),1.02(d,3H,J=6Hz).

[0102] Example 3: Preparation of compound (3)(N-(2-amino-4-methylphenyl)-4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide)

[0103] [ka]

[0104] Step (1): Production of tert-Butyl(5-methyl-2-nitrophenyl)carbamate

[0105] In step (1) of Example 2, 5-methyl-2-nitroaniline was used instead of 4-methyl-2-nitroaniline, and the same procedure was repeated to obtain the target compound, tert-butyl(5-methyl-2-nitrophenyl)carbamate. (Yield 56%)

[0106] Step (2): Production of compound (3-1)

[0107] In step (2) of Example 2, tert-butyl(5-methyl-2-nitrophenyl)carbamate was used instead of tert-butyl(4-methyl-2-nitrophenyl)carbamate, and the same procedure was repeated to obtain compound (3-1) of the target compound. (Yield 97%)

[0108] Step (3): Manufacturing of compound (3-2)

[0109] In step (1) of Example 1, compound (3-1) was used instead of tert-butyl(2-aminophenyl)carbamate, and the same procedure was repeated to obtain compound (3-2), the target compound. (Yield 64%)

[0110] Step (4): Manufacturing of compound (3)

[0111] In step (2) of Example 1, compound (3-2) was used instead of compound (1), and the same procedure was repeated to obtain the target compound (3). (Yield 67%)

[0112] 1H-NMR(300MHz,DMSO-d6)δ11.09(s,1H),8.38(d,1H,J=3Hz),7.74(d,1H,J=3Hz),7.28-7.25(m,1H),6.89(d,1H,J=6Hz),6.64(s,1H),6 .46(d,1H,J=9Hz),4.94(s,2H),4.13-3.42(m,6H),3.32(s,3H),2.41-2.39(m,1H),2.20(s,3H),1.86-1.59(m,3H),1.01(d,3H,J=6Hz)

[0113] Example 4: Preparation of compound (4)(N-(2-amino-4-(dimethylamino)phenyl)-4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide)

[0114] [ka]

[0115] Step (1): Production of tert-Butyl(5-(dimethylamino)-2-nitrophenyl)carbamate

[0116] In step (1) of Example 2, N1,N1-dimethyl-4-nitrobenzene-1,3-diamine was used instead of 4-methyl-2-nitroaniline, and the same procedure was repeated to obtain the target compound, tert-butyl(5-(dimethylamino)-2-nitrophenyl)carbamate. (Yield 52%)

[0117] Step (2): Manufacturing of compound (4-1)

[0118] In step (2) of Example 2, tert-butyl(5-(dimethylamino)-2-nitrophenyl)carbamate was used instead of tert-butyl(4-methyl-2-nitrophenyl)carbamate, and the same procedure was repeated to obtain compound (4-1) of the target compound. (Yield 91%)

[0119] Step (3): Manufacturing of compound (4-2)

[0120] In step (1) of Example 1, compound (4-1) was used instead of tert-butyl(2-aminophenyl)carbamate, and the same procedure was repeated to obtain compound (4-2), the target compound. (Yield 59%)

[0121] Step (4): Production of compound (4)

[0122] In step (2) of Example 1, compound (4-2) was used instead of compound (1), and the same procedure was repeated to obtain the target compound (4). (Yield 77%)

[0123] 1 H-NMR(300MHz,DMSO-d6)δ10.87(s,1H),8.37(d,1H,J=3Hz),7.73(d,1H,J=3Hz),7.10(d,1H,J=9Hz),6.88(d,1H,J=3Hz),6.19(d,1H,J=3Hz),6. 06(dd,1H,12,3Hz),4.84(s,2H),4.20-3.41(m,6H),3.32(s,3H),2.84( s,6H),2.41(m,1H),2.20(s,3H),1.86-1.59(m,3H),1.02(d,3H,J=9Hz).

[0124] Example 5: Preparation of compound (5)(N-(2-amino-5-(dimethylamino)phenyl)-4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide)

[0125] [ka]

[0126] Step (1): Manufacturing of compound (5-2)

[0127] To a toluene (10 mL) solution containing 1 g of compound (5-1) (3.15 mmol, 1 equiv), Me2NH-HCl (1.29 g, 15.7 mmol, 5.00 eq), XPhos (150 mg, 315 μmol, 0.1 eq), t-BuONa (909 mg, 9.46 mmol, 3.00 eq), and Pd2(dba)3 (144 mg, 157 μmol, 0.05 eq) were added. The mixture was stirred at 90°C for 12 hours. After the reaction was complete, water and ethyl acetate were added to the reaction solution and extracted. The organic layer was separated, dried over magnesium sulfate, filtered, and the remaining filtrate was concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 1:100 to 1:0) to obtain the target compound (5-2). (Yield 29%)

[0128] Step (2): Manufacturing of compound (5-3)

[0129] 390 mg of compound (5-2) (1.39 mmol, 1 eq) was dissolved in 3.25 mL of ethanol and 0.65 mL of water. To this mixture, Fe (387 mg, 6.93 mmol, 5.00 eq) and NH4Cl (370 mg, 6.93 mmol, 5.00 eq) were added, and the mixture was stirred at 80°C for 12 hours. After the reaction was complete, water and ethyl acetate were added to the reaction mixture for extraction. The organic layer was separated, dried over magnesium sulfate, filtered, and the remaining filtrate was concentrated under reduced pressure to obtain the target compound (5-3). (Yield 57%)

[0130] Step (3): Manufacturing of compound (5-4)

[0131] In step (1) of Example 1, compound (5-3) was used instead of tert-butyl(2-aminophenyl)carbamate, and the same procedure was repeated to obtain compound (5-4), the target compound. (Yield 71%)

[0132] Step (4): Manufacturing of compound (5)

[0133] In step (2) of Example 1, compounds (5-4) were used instead of compound (1), and the same procedure was repeated to obtain the target compound (5). (Yield 96%)

[0134] 1 H-NMR(400MHz,DMSO-d6)δ11.37-11.33(m,1H),8.42-8.36(m,1H),7.76(d,1H,J=4.2Hz),7.16-7.13(m,1H ),6.94-6.90(m,1H),6.80-6.75(m,1H),6.56-6.51(m,1H),5.01-4.85(m,1H),4.50-4.02(m,5H),3.98(br s,5H),3.43(br d,2H,J=4.4Hz),2.77(s,6H),1.97-1.53(m,2H),1.03(br d,3H,J=7.0Hz).

[0135] Example 6: Preparation of compound (6)(N-(2-amino-4-methoxyphenyl)-4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide)

[0136] [ka]

[0137] Step (1): Manufacturing of compound (6-2)

[0138] In step (1) of Example 1, compound (6-1) was used instead of tert-butyl(2-aminophenyl)carbamate, and the same procedure was repeated to obtain compound (6-2), the target compound. (Yield 92%)

[0139] Step (2): Manufacturing of compound (6)

[0140] In step (2) of Example 1, compound (6-2) was used instead of compound (1), and the same procedure was repeated to obtain the target compound (6). (Yield 6%)

[0141] 1H-NMR(400MHz,DMSO-d6)δ11.07-10.86(m,1H),8.49-8.01(m,1H),7.74(d,1H,J=4.0Hz),7. 31-7.14(m,1H),6.96-6.79(m,1H),6.60-6.38(m,1H),6.22(dd,1H,J=2.4,8.8Hz),5.10(br s,2H),5.00-4.82(m,1H),4.21-4.01(m,2H),3.99-3.74(m,2H),3.71(m,4H),3.43(br dd,1H,J=3.2,5.4Hz),3.33-3.32(m,2H),3.32-3.26(m,1H),2.46-2.37(m,1H),1.93-1.69(m,1H),1.68-1.53(m,1H),1.03(d,3H,J=7.2Hz).

[0142] Example 7: Preparation of compound (7)(N-(2-amino-5-methoxyphenyl)-4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide)

[0143] [ka]

[0144] Step (1): Manufacturing of compound (7-2)

[0145] In step (1) of Example 1, compound (7-1) was used instead of tert-butyl(2-aminophenyl)carbamate, and the same procedure was repeated to obtain compound (7-2), the target compound.

[0146] Step (2): Production of compound (7)

[0147] In step (2) of Example 1, compound (7-2) was used instead of compound (1), and the same procedure was repeated to obtain the target compound (7). (Yield 15%)

[0148] 1 H-NMR(400MHz,DMSO-d6)δ11.50-11.31(m,1H),8.43-8.35(m,1H),7.76(d,1H,J=4.0Hz),7.35-7.23(m,1H),6.91(br s,1H),6.81(brd,1H,J=8.6Hz),6.66-6.49(m,1H),4.91(brd,1H,J=3.2Hz),4.56(br s,2H),4.23-4.05(m,2H),3.96(br dd,1H,J=3.6,12.4Hz),3.88-3.58(m,5H),3.45-3.40(m,1H),3.33(br s,2H),3.28(br d,1H,J=4.0Hz),2.40(br dd,1H,J=5.6,12.0Hz),1.89-1.52(m,2H),1.03(br d,3H,J=7.2Hz).

[0149] Example 8: Preparation of compound (8)(N-(2-amino-4-hydroxyphenyl)-4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide)

[0150] [ka]

[0151] Step (1): Production of compound (8)

[0152] 169 mg (0.676 mmol, 3.00 eq) of compound (6-2) (0.225 mmol, 1 equiv) was dissolved in a 1.3 mL solution of dichloromethane, to which BBr3 was added. The mixture was stirred at 0°C for 4 hours. After the reaction was complete, the reaction solution was concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography to obtain the target compound (8). (Yield 30%)

[0153] 1 H-NMR(400MHz,DMSO-d6)δ10.91-10.85(m,1H),9.11-8.97(m,1H),8.38-8.35(m,1H),7.72(d,1H,J=3.6Hz),7.04(br d,1H,J=8.4Hz),6.88(br d,1H,J=2.8Hz),6.28-6.21(m,1H),6.04(br d,1H,J=8.8Hz),5.01-4.92(m,2H),4.21-4.05(m,2H),4.02-3.92(m,1H),3.85-3.64(m,3H),3.51-3.47(m,2H),3.24-3.20(m,1H),2.38(br d,2H,J=2.4Hz),1.90-1.68(m,1H),1.66-1.54(m,1H),1.16-0.98(m,3H).

[0154] Example 9: Preparation of compound (9)(N-(2-amino-5-hydroxyphenyl)-4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide)

[0155] [ka]

[0156] Step (1): Production of compound (9)

[0157] In step (1) of Example 8, compound (7-2) was used instead of compound (6-2), and the same procedure was repeated to obtain the target compound (9). (Yield 12%)

[0158] 1 H-NMR(400MHz,DMSO-d6)δ11.39(br d,1H,J=8.0Hz),8.71(s,1H),8.40-8.36(m,1H),7.76(d,1H,J=4.0Hz),7.23-7.13(m,1H),6.94-6.87(m,1H),6.70(d,1H,J=8.4Hz),6.43(dd, 1H,J=2.8,8.4Hz),4.98-4.83(m,1H),4.45-4.30(m,2H),4.21-4.05(m ,2H),4.03-3.91(m,1H),3.86-3.65(m,2H),3.47-3.42(m,2H),3.22(br d,1H,J=7.2Hz),2.42(br d,2H,J=8.2Hz),2.09-1.37(m,2H),1.24(s,1H),1.14-1.13(m,1H),1.02(br d,3H,J=6.8Hz).

[0159] Example 10: Preparation of compound (10)(N-(6-amino-1,3-dimethyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide)

[0160] [ka]

[0161] Step (1): Preparation of tert-butyl(1,3-dimethyl-6-nitro-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)carbamate

[0162] In step (1) of Example 2, 5-amino-1,3-dimethyl-6-nitro-1,3-dihydro-2H-benzo[d]imidazol-2-one was used instead of 4-methyl-2-nitroaniline, and the same procedure was repeated to obtain the target compound, tert-butyl(1,3-dimethyl-6-nitro-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)carbamate.

[0163] Step (2): Production of compound (10-1)

[0164] In step (2) of Example 2, tert-butyl(1,3-dimethyl-6-nitro-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)carbamate was used instead of tert-butyl(4-methyl-2-nitrophenyl)carbamate, and the same procedure was repeated to obtain compound (10-1) of the target compound. (Yield 35%)

[0165] Step (3): Production of compound (10⁻²)

[0166] In step (1) of Example 1, compound (10-1) was used instead of tert-butyl(2-aminophenyl)carbamate, and the same procedure was repeated to obtain the target compound (10-2). (Yield 42%)

[0167] Step (4): Manufacturing of compound (10)

[0168] In step (2) of Example 1, compound (10-2) was used instead of compound (1), and the same procedure was repeated to obtain the target compound (10). (Yield 61%)

[0169] 1 H-NMR(300MHz,DMSO-d6)δ11.19-11.17(m,1H),8.39-8.37(d,1H,J=6Hz),7.74(d, 1H,J=2.1Hz),7.24-7.24(m,1H),6.90(s,1H),6.62-6.61(d,1H,J=2.1Hz),4.91(s, 1H),4.81-4.79(m,2H),4.17-3.94(m,4H),3.83-3.70(m,2H),3.47-3.36(m,3H),3. 25-3.25(m,5H),2.44-2.38(m,1H),1.88-1.59(m,3H),1.03-1.02(d,3H,J=3.6Hz).

[0170] Example 11: Preparation of compound (11)(N-(2-aminophenyl)-N-((2-aminophenyl)carbamoyl)-4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide

[0171] [ka]

[0172] Step (1): Production of compound (11-2)

[0173] To a 30 mL solution of dioxane containing 1 g of tofacitinib (3.20 mmol, 1 equiv), compound (11-1) (0.63 g, 3.84 mmol, 1.2 eq) and triethylamine (421 mg, 4.16 mmol, 1.3 eq) were added. The mixture was stirred at 120°C for 2 days. After the reaction was complete, the reaction solution was concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (methanol:dichloromethane = 1:10) to obtain the target compound (11-2). (Yield 60%)

[0174] Step (2): Production of compound (11-3)

[0175] 2.9 g of compound (11-2) (6.09 mmol, 1 equiv) was dissolved in 50 mL of THF, and the mixture was cooled to 0°C. 219 mg of NaH (60% content) (7.31 mmol, 1.2 equiv) was added, and the mixture was stirred for 30 minutes. The mixture was then heated to room temperature and stirred for 18 hours. After the reaction was complete, water and ethyl acetate were added to the reaction mixture and extracted. The organic layer was separated, dried over magnesium sulfate, filtered, and the remaining filtrate was concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (methanol:dichloromethane = 1:10) to obtain the target compound (11-3). (Yield 51%)

[0176] Step (3): Manufacturing of compound (11)

[0177] 1.1 g of compound (11-3) (1.72 mmol, 1 equiv) was dissolved in methanol (50 mL), to which 100 mg of 10% Pd / C (10 wt.) was added. The mixture was then stirred under atmospheric pressure and hydrogen gas for 18 hours. The mixture was then heated to room temperature and stirred for 18 hours. After the reaction was complete, the reaction mixture was passed through celite and filtered, and washed with 50 mL of methanol. The filtrate was concentrated under reduced pressure, and the concentrate was purified by silica gel column chromatography (methanol:dichloromethane = 1:10) to obtain the target compound (11). (Yield 6%)

[0178] 1 H-NMR(400MHz,DMSO-d6)δ11.24-11.19(m,1H),8.40-8.34(m,1H),7.73(dd,1H,J=11.2,4.0H z),7.43(dd,1H,J=8.0,1.2Hz),7.08-6.81(m,5H),6.75-6.69(m,1H),6.65-6.62(m,1H),6.5 8-6.51(m,1H),5.58(d,1H,J=8.4Hz),5.02(br,4H),4.00-3.79(m,2H),3.75-3.55(m,2H),3. 40-3.32(m,5H),2.46-2.32(m,1H),1.94-1.74(m,1H),1.72-1.64(m,1H),1.09-0.99(m,3H).

[0179] Example 12: Preparation of compound (12)(N-(2-amino-4-methylphenyl)-N-((2-amino-4-methylphenyl)carbamoyl)-4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide

[0180] [ka]

[0181] Step (1): Preparation of compound (12-1)

[0182] 500 mg of 4-methyl-2-nitroaniline (3.29 mmol, 1 equiv) was dissolved in 28 mL of 1,2-dichloroethane, and the mixture was cooled to 0-5°C. 448 mg of triphosgene (1.64 mmol, 0.5 equiv) was dissolved in 66 mL of 1,2-dichloroethane and added, and the temperature was raised to room temperature. Subsequently, 458 μL of trimethylamine (3.29 mmol, 1 equiv) was added, and the mixture was refluxed and stirred for 2-16 hours. After the reaction was complete, the temperature was lowered, and the mixture was used for the next reaction without purification.

[0183] Step (2): Preparation of compound (12-2)

[0184] Compound (12-1) (3.29 mmol, 2 equivs) was treated with 513 mg of tofacitinib (1.64 mmol, 1 equiv) and 265 μL of pyridine (3.29 mmol, 2 equivs), followed by reflux stirring for 1-2 hours. After the reaction was complete, the mixture was concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (ethyl acetate:hexane = 1:1) to obtain the target compound (12-2). (Yield 85%)

[0185] Step (3): Production of compound (12-3)

[0186] 100 mg of compound (12-2) (0.20 mmol, 1 equiv) was dissolved in 2.6 mL of anhydrous THF, and the mixture was cooled to 0-5°C. 410 μL of 1 M LiHMDS in THF (0.41 mmol, 2 equiv) was added, and the temperature was raised to room temperature. Compound (12-1) was added, and the mixture was stirred for 1 hour. After the reaction was complete, water and ethyl acetate were added to the reaction mixture and extracted. The organic layer was separated, dried over magnesium sulfate, filtered, and the remaining filtrate was concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (dichloromethane:methanol = 10:1) to obtain the target compound (12-3). (Yield 40%)

[0187] Step (4): Manufacturing of compound (12)

[0188] 54 mg of compound (12-3) (0.08 mmol, 1 equiv) was dissolved in 5 mL of ethanol, 1.5 mL of THF, and 1.5 mL of water. At room temperature, 45 mg of Fe (0.81 mmol, 10 equiv) and 44 mg of NH4Cl (0.81 mmol, 10 equiv) were added to the mixture and the mixture was stirred under reflux for 1 hour. After the reaction was complete, ethyl acetate was added to the reaction mixture and extracted. The organic layer was separated, dried over magnesium sulfate, filtered, and the remaining filtrate was concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (dichloromethane:methanol = 10:1) to obtain the target compound (12). (Yield 38%)

[0189] 1 H-NMR(300MHz,DMSO-d6)δ11.56-11.53,11.12-11.09(m,1H),10.46-10.42 (m,1H),8.16-8.13(d,1H,J=9Hz),7.28-7.25(d,1H,J=9Hz),6.93-6.87(m, 2H),6.80-6.71(m,1H),6.63-6.30(m,6H),5.55-5.53(m,1H),4.93(s,4H), 4.00-3.51(m,7H),2.29-2.27(m,6H),1.73-1.58(m,2H),1.05-1.03(m,3H).

[0190] Example 13: Preparation of compound (13)(N-(2-amino-4-methoxyphenyl)-N-((2-amino-4-methoxyphenyl)carbamoyl)-4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide):(N-(2-amino-4-methoxyphenyl)-N-((2-amino-4-methoxyphenyl)carbamoyl)-4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide)

[0191] [ka]

[0192] Step (1): Production of compound (13-1)

[0193] In step (1) of Example 12, 4-methoxy-2-nitroaniline was used instead of 4-methyl-2-nitroaniline, and the same procedure was repeated to obtain compound (13-1) of the target compound.

[0194] Step (2): Preparation of compound (13-2)

[0195] In step (2) of Example 12, compound (13-1) was used instead of compound (12-1), and the same procedure was repeated to obtain the target compound (13-2). (Yield 88%)

[0196] Step (3): Production of compound (13-3)

[0197] In step (3) of Example 12, compound (13-2) was used instead of compound (12-2), and the same procedure was repeated to obtain compound (13-3), the target compound, without purification.

[0198] Step (4): Manufacturing of compound (13)

[0199] In step (4) of Example 12, compound (13-3) was used instead of compound (12-3), and the same procedure was repeated to obtain the target compound (13). (Yield 27%)

[0200] 1H-NMR(300MHz,DMSO-d6)δ10.97-10.94(m,1H),10.49-10.36(m,1H),8.39-8. 36(m,1H),7.75-7.71(m,1H),7.21-7.18(d,1H,J=9Hz),6.93-6.85(m,1H),6.8 1-6.78(d,1H,J=9Hz),6.52-6.49(m,2H),6.38-6.11(m,4H),5.54-5.50(m,1H) ,5.08-5.02(m,4H),3.96-3.61(m,13H),1.76-1.66(m,2H),1.05-1.01(m,3H).

[0201] Example 14: Preparation of compound (14)(N-(2-(3-(2-aminophenyl)-1-methylureido)ethyl)-4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide)

[0202] [ka]

[0203] Step (1): Production of compound (14-1)

[0204] 5 g of tofacitinib (16.0 mmol, 1 equiv) and 3.24 g of TEA (32.0 mmol, 2 equivs) were dissolved in dichloromethane. 5.84 g of bis(4-nitrophenyl)carbonate (19.2 mmol, 1.2 equivs) was added, and the mixture was stirred at 40°C for 2 hours. 4.54 g of tert-butyl methyl(2-(methylamino)ethyl)carbamate (24.0 mmol, 1.5 eq) was added, and the mixture was stirred at 40°C for 16 hours. After the reaction was complete, water and dichloromethane were added to the reaction mixture for extraction. The organic layer was separated, dried over magnesium sulfate, filtered, and the remaining filtrate was concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (dichloromethane:methanol = 20:1) to obtain compound (14-1), the target compound. (Yield 39%)

[0205] Step (2): Preparation of compound (14-2)

[0206] 3.3 g of compound (14-1) (7.7 mmol, 1 equiv) was dissolved in 30 mL of ethyl acetate and then cooled to 0-5°C. Hydrochloric acid gas was added to the mixture for 2 minutes. After the reaction was complete, the reaction solution was filtered to obtain the target compound (14-2).

[0207] Step (3): Production of compound (14-3)

[0208] 900 mg of compound (14-2) (2.1 mmol, 1 equiv) and 320 mg of TEA (3.2 mmol, 1.5 equiv) were dissolved in 20 mL of dichloromethane. 415 mg of 1-isocyanato-2-nitrobenzene (2.5 mmol, 1.2 equiv) was added, and the mixture was stirred at room temperature for 16 hours. After the reaction was complete, water and dichloromethane were added to the reaction mixture for extraction. The organic layer was separated, dried over magnesium sulfate, filtered, and the remaining filtrate was concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (dichloromethane:methanol = 20:1) to obtain the target compound (14-3). (Yield 36%)

[0209] Step (4): Manufacturing of compound (14)

[0210] 445 mg of compound (14-3) (0.75 mmol, 1 equiv) was dissolved in 10 mL of methanol. At room temperature, 80 mg of 10% Pd / C was added to the mixture and the mixture was stirred under atmospheric pressure and hydrogen gas for 2 hours. After the reaction was complete, the mixture was purified using a Celite filter and washed with 20 mL of methanol. The filtrate was concentrated under reduced pressure, and the concentrate was purified by silica gel column chromatography (dichloromethane:methanol = 20:1) to obtain the target compound (14). (Yield 36%)

[0211] 1 H-NMR(400MHz,DMSO-d6)δ8.25-8.18(m,1H),7.73-7.50(m,1H),7.40-7.20(m,1H) ,7.06-6.82(m,2H),6.80-6.65(m,2H),6.51(d,1H,J=7.2Hz),4.95-4.65(m,3H),4. 20-4.00(m,2H),3.99-3.35(m,8H),3.27(s,3H),3.16-2.98(m,3H),2.96-2.62(m,3 H),2.46-2.31(m,1H),1.89-1.68(m,1H),1.63-1.52(m,1H),1.01(d,3H,J=7.2Hz).

[0212] Example 15: Preparation of compound (15)(N-(2-(3-(2-amino-4-methylphenyl)-1-methylureido)ethyl)-4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide

[0213] [ka]

[0214] Step (1): Production of compound (15-1)

[0215] 250 mg of compound (14-1) (0.47 mmol, 1 equiv) was dissolved in 20 mL of dichloromethane. 5 mL of TFA (20V) was added, and the mixture was stirred at room temperature for 2 hours. After the reaction was complete, the mixture was concentrated under reduced pressure to obtain compound (15-1). (Yield 99%)

[0216] Step (2): Production of compound (15-2)

[0217] 202 mg of compound (15-1) (0.47 mmol, 1 equiv) and 84 mg of compound (12-1) were dissolved in 40 mL of anhydrous THF. 47 mg of TEA (0.47 mmol, 1 equiv) was added, and the mixture was stirred at room temperature for 16 hours. After the reaction was complete, the concentrated solution was purified by silica gel column chromatography (dichloromethane:methanol = 20:1) to obtain the target compound (15-2). (Yield 42%)

[0218] Step (3): Manufacturing of compound (15)

[0219] 120 mg of compound (15-2) (0.20 mmol, 1 equiv) was dissolved in 15 mL of methanol. At room temperature, 100 mg of 10% Pd / C was added to the mixture and the mixture was stirred under atmospheric pressure and hydrogen gas for 1 hour. After the reaction was complete, the mixture was filtered through a Celite filter, and the filtrate was concentrated under reduced pressure. The concentrate was purified by reverse-phase prep-HPLC to obtain the target compound (15). (Yield 40%)

[0220] 1H-NMR(400MHz, DMSO-d6) δ 8.22 - 8.20 (m, 1H), 7.56 - 7.53 (m, 1H), 7.30 (s, 1H), 6.86 - 6.76 (m, 2H), 6.52 (s, 1H), 6.34 - 6.62 (d, 1H, J = 7.2Hz), 4.85 (s, 1H), 4.65 (s, 2H), 4.17 - 3.91 (m, 3H), 3.78 - 3.63 (m, 4h), 3.46 - 3.37 (m, 3H), 3.27 (s, 4H), 3.11 - 3.06 (m, 1H), 2.88 (s, 1H), 2.67 - 2.63 (m, 1H), 2.39 - 2.36 (m, 1H), 2.15 (s, 3H), 1 - 84 - 1.68 (m, 1H), 1.62 - 1.57 (m, 1H), 1.02 - 1.00 (d, 3H, J = 7.2Hz).

[0221] Example 16: Preparation of Compound (16) (N-(2-(3-(2-amino-5-methylphenyl)-1-methylureido)ethyl)-4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide)

[0222]

Chemical Structure

[0223] Step (1): Preparation of Compound (16-1)

[0224] 152 mg of 5-methyl-2-nitroaniline (1.0 mmol, 1 equiv) was dissolved in 10 mL of 1,2-dichloroethane, and the mixture was cooled to 0-5°C. 148 mg of triphosgene (0.5 mmol, 0.5 equiv) was dissolved in 20 mL of 1,2-dichloroethane and added, and the temperature was raised to room temperature. The mixture was then stirred under reflux for 16 hours. After the reaction was complete, the mixture was concentrated under reduced pressure to obtain compound (16-1). (Yield 96%)

[0225] Step (2): Preparation of compound (16-2)

[0226] In step (2) of Example 15, compound (16-1) was used instead of compound (12-1), and the same procedure was repeated to obtain the target compound (16-2). (Yield 58%)

[0227] Step (3): Manufacturing of compound (16)

[0228] In step (3) of Example 15, compound (16-2) was used instead of compound (15-2), and the same procedure was repeated to obtain the target compound (16). (Yield 58%)

[0229] 1 H-NMR(400MHz,DMSO-d6)δ8.22-8.21(m,1H),7.60(s,1H),7.34-7.29(m,1H),6.82-6.76(m,2H),6.7 1-6.69(d,1H,J=8Hz),6.63-6.61(d,1H,J=8Hz),4.85(s,1H),4.53(s,2H),4.16-3.91(m,3H),3.86- 3.63(m,4H),3.55-3.40(m,3H),3.27(s,4H),3.22-2.97(m,3H),2.89-2.67(m,1H),2.63-2.50(m,1H) ),2.39-2.36(m,1H),2.13(s,3H),1.86-1.71(m,1H),1.61-1.57(m,1H),1.02-1.00(d,3H,J=7.2Hz).

[0230] Example 17: Preparation of compound (17)(N-(2-(3-(2-amino-4-methoxyphenyl)-1-methylureido)ethyl)-4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide)

[0231] [ka]

[0232] Step (1): Production of compound (17-1)

[0233] 1.2 g of compound (14-2) (2.67 mmol, 1 equiv) and 540 mg of TEA (5.34 mmol, 2 equivs) were dissolved in 10 mL of anhydrous THF, and the mixture was cooled to 0-5°C. Compound (13-1) (2.67 mmol, 1 equiv) was added, and the mixture was stirred at room temperature for 16 hours. After the reaction was complete, the reaction mixture was concentrated under reduced pressure, and the concentrate was purified by silica gel column chromatography (dichloromethane:methanol = 25:1) to obtain the target compound (17-1). (Yield 24%)

[0234] Step (2): Manufacturing of compound (17)

[0235] 400 mg of compound (17-1) (0.64 mmol, 1 equiv) was dissolved in 20 mL of methanol. At room temperature, 80 mg of 10% Pd / C was added to the mixture and the mixture was stirred under atmospheric pressure and hydrogen gas for 2 hours. After the reaction was complete, the mixture was filtered through a Celite filter, and the filtrate was concentrated under reduced pressure. The concentrate was purified by reverse-phase prep-HPLC to obtain the target compound (17). (Yield 29%)

[0236] 1 H-NMR(400MHz,DMSO-d6)δ8.21(d,1H,J=5.6Hz),7.51(s,1H),7.30(s,1H),6.90-6.6 6(m,2H),6.29(d,1H,J=2.7Hz),6.10(dd,1H,J=8.4,2.1Hz),4.94-4.66(m,3H),4.20- 3.53(m,10H),3.48-3.35(m,3H),3.27(s,3H),3.17-2.98(m,3H),2.88(s,2H),2.64(s ,1H),2.45-2.31(m,1H),1.88-1.66(m,1H),1.64-1.51(m,1H),1.02(d,3H,J=7.2Hz).

[0237] Example 18: Preparation of compound (18)(N-(2-(3-(2-amino-5-methoxyphenyl)-1-methylureido)ethyl)-4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide)

[0238] [ka]

[0239] Step (1): Preparation of Compound (18-1)

[0240] Dissolve 500 mg of 5-methoxyl-2-nitroaniline (2.97 mmol, 1 equiv) in 10 mL of ethyl acetate, and cool the mixture to 0 - 5 °C. Add 360 mg of TEA (3.57 mmol, 10.2 eq) and 291 mg of triphosgene (0.98 mmol, 0.33 equiv), and stir at room temperature for 1 hour. After the reaction is completed, use it in the next reaction without purification.

[0241] Step (2): Preparation of Compound (18-2)

[0242] In step (1) of Example 17 above, use Compound (18-1) instead of Compound (13-1), and repeat the same procedure to obtain Compound (18-2) of the target compound. (Yield 14%)

[0243] Step (3): Preparation of Compound (18)

[0244] In step (2) of Example 17 above, use Compound (18-2) instead of Compound (17-1), and repeat the same procedure to obtain Compound (18) of the target compound. (Yield 49%)

[0245] 1 H-NMR (400 MHz, DMSO-d6) δ 8.26 - 8.16 (m, 1H), 7.69 (s, 1H), 7.45 - 7.19 (m, 1H), 6.85 - 6.60 (m, 3H), 6.56 - 6.48 (m, 1H), 4.96 - 4.76 (m, 1H), 4.57 (s, 2H), 4.20 - 3.50 (m, 10H), 3.50 - 3.35 (m, 3H), 3.30 - 3.20 (m, 3H), 3.18 - 3.00 (m, 3H), 2.96 - 2.78 (m, 2H), 2.74 - 2.56 (m, 1H), 2.44 - 2.31 (m, 1H), 1.90 - 1.65 (m, 1H), 1.64 - 1.50 (m, 1H), 1.01 (d, 3H, J = 8 Hz).

[0246] Example 19: Preparation of compound (19)(N-(2-(3-(2-amino-4-hydroxyphenyl)-1-methylureido)ethyl)-4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide)

[0247] [ka]

[0248] Step (1): Production of compound (19-1)

[0249] 1 g of 4-amino-3-nitrophenol (6.49 mmol, 1 equiv) was dissolved in 10 mL of DMF, and the mixture was cooled to 0-5°C. 801 mg of t-BuOK (7.14 mmol, 1.1 equiv) was dissolved in 5 mL of THF and added, and the mixture was stirred at 0°C for 20 minutes. Subsequently, 1.2 g of benzyl bromide (7.14 mmol, 1.1 equiv) was dissolved in 5 mL of DMF and added dropwise, and the mixture was stirred at 0°C for 20 minutes. After the reaction was complete, 20 mL of aqueous NH4Cl solution was added dropwise, and then ethyl acetate was added for extraction. After separating the organic layer, it was dried over magnesium sulfate and filtered, and the remaining filtrate was concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (dichloromethane:methanol = 20:1) to obtain compound (19-1), the target compound. (Yield 76%)

[0250] Step (2): Preparation of compound (19-2)

[0251] In step (1) of Example 18, compound (19-1) was used instead of 5-methoxyl-2-nitroaniline, and the same procedure was repeated to obtain compound (19-2), the target compound.

[0252] Step (3): Production of compound (19-3)

[0253] In step (1) of Example 17, compound (19-2) was used instead of compound (13-1), and the same procedure was repeated to obtain the target compound (19-3). (Yield 11%)

[0254] Step (4): Manufacturing of compound (19)

[0255] In step (2) of Example 17, compound (19-3) was used instead of compound (17-1), and the same procedure was repeated to obtain the target compound (19). (Yield 57%)

[0256] 1 H-NMR(400MHz,DMSO-d6)δ8.82-8.75(m,1H),8.25-8.16(m,1H),7.49-7.21(m,2H),6.85-6. 50(m,2H),6.17-6.09(m,1H),5.98-5.88(m,1H),4.96-4.76(m,1H),4.63(s,2H),4.20-3.50 (m,7H),3.49-3.34(m,3H),3.30-3.18(m,3H),3.16-2.95(m,3H),2.94-2.77(m,2H),2.71-2 .54(m,1H),2.45-2.31(m,1H),1.91-1.65(m,1H),1.64-1.50(m,1H),1.02(d,3H,J=8.0Hz).

[0257] Example 20: Preparation of compound (20)(N-(2-(3-(2-amino-5-hydroxyphenyl)-1-methylureido)ethyl)-4-(((1S,2R)-5-(2-cyanoacetyl)-2-methylcyclohexyl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide)

[0258] [ka]

[0259] Step (1): Production of compound (20-1)

[0260] 1.5 g of 5-fluoro-2-nitroaniline (9.6 mmol, 1 equiv) was dissolved in 2.6 g of K2CO3 and 40 mL of n-butanol, and the mixture was stirred under reflux at 100°C for 16 hours. After the reaction was complete, the mixture was cooled to room temperature, filtered, and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (PE:ethyl acetate = 3:1) to obtain compound (20-1), the target compound. (Yield 23%)

[0261] Step (2): Production of compound (20-2)

[0262] In step (1) of Example 18, compound (20-1) was used instead of 5-methoxyl-2-nitroaniline, and the same procedure was repeated to obtain the target compound (19-2).

[0263] Step (3): Production of compound (20-3)

[0264] In step (1) of Example 17, compound (20-2) was used instead of compound (13-1), and the same procedure was repeated to obtain the target compound (20-3). (Yield 13%)

[0265] Step (4): Manufacturing of compound (20)

[0266] In step (2) of Example 17, compound (20-3) was used instead of compound (17-1), and the same procedure was repeated to obtain the target compound (20). (Yield 45%)

[0267] 1 H-NMR(400MHz,DMSO-d6)δ8.48(s,1H),8.21(d,1H,J=5.6Hz),7.61(s,1H),7.42-7. 22(m,1H),6.81-6.52(m,3H),6.33(dd,1H,J=8.4,2.4Hz),4.84(s,1H),4.56(s,2H), 4.20-3.51(m,7H),3.40(s,3H),3.27(s,3H),3.09(d,3H,J=21.2Hz),2.88(s,2H),2. 73-2.56(m,1H),2.37(s,1H),1.90-1.66(m,1H),1.57(s,1H),1.01(d,3H,J=7.2Hz).

[0268] Example 21: Preparation of compound (21)(N-(2-(3-(2-amino-5-(dimethylamino)phenyl)-1-methylureido)ethyl)-4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide)

[0269] [ka]

[0270] Step (1): Production of compound (21-1)

[0271] 181 mg of N1,N1-dimethyl-4-nitrobenzene-1,3-diamine (1.0 mmol, 1 equiv) was dissolved in 10 mL of 1,2-dichloroethane, and the mixture was cooled to 0-5°C. 148 mg of triphosgene (0.5 mmol, 0.5 equiv) was dissolved in 20 mL of 1,2-dichloroethane and added, and the temperature was raised to room temperature. The mixture was then stirred under reflux for 16 hours. After the reaction was complete, the mixture was concentrated under reduced pressure to obtain compound (21-1). (Yield 97%)

[0272] Step (2): Preparation of compound (21-2)

[0273] In step (2) of Example 15, compound (21-1) was used instead of compound (12-1), and the same procedure was repeated to obtain the target compound (21-2) (yield 76%).

[0274] Step (3): Manufacturing of compound (21)

[0275] 110 mg of compound (21-2) (0.17 mmol, 1 equiv) was dissolved in 10 mL of ethanol and 3 mL of water. At room temperature, 95 mg of Fe (1.7 mmol, 10 equiv) and 91 mg of NH4Cl (1.7 mmol, 10 equiv) were added to the mixture and the mixture was stirred under reflux for 1 hour. After the reaction was complete, the mixture was filtered through a Celite filter, and the filtrate was concentrated under reduced pressure. The concentrate was purified by reverse-phase prep-HPLC to obtain the target compound (21). (Yield 34%)

[0276] 1H-NMR(400MHz,DMSO-d6)δ8.22-8.19(m,1H),7.67(s,1H),7.39-7.28(m,1H),6.74-6.74(m,1H),6 .65-6.63(m,2H),6.45-6.42(dd,1H,J=8.8Hz,2.8Hz),5.59-5.21(m,2H),4.85(s,1H),4.17-3.91 (m,3H),3.86-3.63(m,4H),3.54-3.31(m,3H),3.26(s,3H),3.12-3.07(m,3H),2.87-2.75(m,2H), 2.71(s,6H),2.67-2.63(m,1H),2.49-2.33(m,1H),1.86-1.57(m,2H),1.02-1.00(d,3H,J=6.8Hz).

[0277] Example 22: Compound (22)(N-(2-(3-(6-amino-1,3-dimethyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazole-5-yl)-1-methylureido)ethyl)-4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidine-3-yl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide:N-(2-(3-(6-amino-1,3 -dimethyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-1-methylureido)ethyl)-4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide)

[0278] [ka]

[0279] Step (1): Preparation of compound (22-1)

[0280] In step (2) of Example 15, 5-isocyanato-1,3-dimethyl-6-nitro-1H-benzo[d]imidazol-2(3H)-one was used instead of compound (12-1), and the same procedure was repeated to obtain the target compound (22-1). (Yield 70%)

[0281] Step (2): Production of compound (22)

[0282] In step (3) of Example 21, compound (22-1) was used instead of compound (21-2), and the same procedure was repeated to obtain the target compound (22). (Yield 13%)

[0283] 1 H-NMR(400MHz,DMSO-d6)δ8.21-8.21(m,1H),7.66(s,1H),7.38-7.28(m,1 H),6.77(s,2H),6.51(s,1H),4.88-4.84(m,1H),4.55-4.50(m,2H),4.18-3 .62(m,7H),3.45-3.40(m,3H),3.26-3.05(m,12H),2.89(s,2H),2.66-2.61 (m,1H),2.41-2.35(m,1H),1.85-1.54(m,2H),1.02-1.00(d,3H,J=7.2Hz).

[0284] Example 23: Compound (23)((4-((((2-(3-(2-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamido)ethyl)-3-methylureido)-4-(dimethylamino)phenyl)carbamoyl)oxy)methyl)phenyl)boronic (acid) manufacturing

[0285] [ka]

[0286] Step (1): Production of compound (23-1)

[0287] 603 mg of compound (21) (1 mmol, 1 equiv) and 296 mg of 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl carbonochloridate (1 mmol, 1 equiv) were dissolved in 40 mL of dichloromethane. 303 mg of TEA (3 mmol, 3 equiv) was added, and the mixture was stirred at room temperature for 16 hours. After the reaction was complete, water and ethyl acetate were added and the mixture was extracted. The organic layer was separated, dried over magnesium sulfate, filtered, and the remaining filtrate was concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (dichloromethane:methanol = 20:1) to obtain the target compound (23-1). (Yield 46%)

[0288] Step (2): Manufacturing of compound (23)

[0289] 400 mg of compound (23-1) (0.46 mmol, 1 equiv) was dissolved in 20 mL of MeCN. 10 mg of NH4HCO3 (0.79 mmol, 1.7 equiv) was dissolved in 10 mLH of water and added to the mixture, and the mixture was stirred for 16 hours. After the reaction was complete, water and ethyl acetate were added and the mixture was extracted. After separating the organic layer, it was dried over magnesium sulfate and filtered. The remaining filtrate was concentrated under reduced pressure. The concentrate was purified by reverse-phase prep-HPLC to obtain the target compound (23). (Yield 30%)

[0290] 1 H-NMR(400MHz,DMSO-d6)δ8.72-8.72(m,1H),8.20-8.19(m,1H),8.06(s,2H),7.89-7.74(m,3H),7. 32-7.13(m,4H),6.84-6.84(m,1H),6.70-6.69(m,1H),6.50-6.48(d,1H,J=9.2Hz),5.11-5.02(m,2 H),4.86-4.81(m,1H),4.17-3.86(m,3H),3.76-3.55(m,4H),3.42-3.39(m,2H),3.31-2.87(m,7H), 2.84(s,8H),2.80-2.59(m,1H),2.39-2.32(m,1H),1.83-1.55(m,2H),1.02-1.00(d,3H,J=7.2Hz).

[0291] Example 24: Compound (24)((4-((((6-(3-(2-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidine-3-yl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide)ethyl)-3-methylureido)-1,3-dimethyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazole-5-yl)carbamoyl)oxy)methyl)phenyl)boric acid:(4-((((6-(3-(2-(4-(((3R ,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamido)e thyl)-3-methylureido)-1,3-dimethyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)carbamoyl)oxy)methyl)phenyl)boronic production of acid)

[0292] [ka]

[0293] Step (1): Production of compound (24-1)

[0294] In step (1) of Example 23, compound (22) was used instead of compound (21), and the same procedure was repeated to obtain the target compound (24-1). (Yield 29%)

[0295] Step (2): Manufacturing of compound (24)

[0296] In step (2) of Example 23, compound (24-1) was used instead of compound (23-1), and the same procedure was repeated to obtain the target compound (24). (Yield 30%)

[0297] 1H-NMR(400MHz,DMSO-d6)δ8.92-8.92(m,1H),8.20-8.19(m,1H),8.07(s,2H),8.00-7.63( m,3H),7.38-7.22(m,4H),7.10(s,1H),5.16-5.05(m,2H),4.87-4.76(m,1H),4.17-3.90( m,3H),3.76-3.57(m,4H),3.44-3.38(m,2H),3.29-3.20(m,10H),3.09-2.99(m,3H),2.86 -2.82(m,2H),2.67-2.61(m,1H),2.39-2.33(m,3H),1.85-1.51(m,2H),0.99-0.98(m,3H).

[0298] Example 25: Preparation of compound (25) (4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl(2-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamido)phenyl)carbamate

[0299] [ka]

[0300] Step (1): Production of compound (25-2)

[0301] 4.6 g of Na2CO3 (43 mmol, 10 eq.) was dissolved in 5 mL of toluene, and the mixture was cooled to 0°C. 2.5 g of triphosgene (8.4 mmol, 1.95 eq.) was added, and the mixture was stirred for 30 minutes. 5 mL of toluene containing 1 g of compound (25-1) (4.3 mmol, 1 eq.) was added dropwise to the reaction mixture, and the mixture was heated to room temperature and stirred for 16 hours. After the reaction was complete, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 1:20) to obtain the target compound (25-2). (Yield 28%)

[0302] Step (2): Manufacturing of compound (25)

[0303] Compound (25-2) (160 mg, 0.54 mmol, 1.2 eq) was added to a 5 mL THF solution containing 200 mg of compound (1) (0.45 mmol, 1 equiv). The mixture was stirred at room temperature for 2 hours. After the reaction was complete, the reaction solution was concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography to obtain the target compound (25). (Yield 22%)

[0304] 1 H-NMR(400MHz,CDCl3)δ11.94-11.75(m,1H),8.24-8.23(m,1H),7.71-7.67 (m,5H),7.30-7.19(m,3H),7.17-7.15(m,2H),6.58-6.55(m,1H),5.14(s,2 H),5.07(br,1H),4.03-3.72(m,2H),3.56-3.42(m,4H),3.33-3.31(m,3H), 2.48-2.36(m,1H),1.94-1.52(m,2H),1.27(s,12H),1.01(d,3H,J=6.8Hz).

[0305] Example 26: Preparation of compound (26) (4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl(2-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamido)-4-methylphenyl)carbamate

[0306] [ka]

[0307] Step (1): Manufacturing of compound (26)

[0308] In step (2) of Example 25, compound (2) was used instead of compound (1), and the same procedure was repeated to obtain compound (26), the target compound. (Yield 18%)

[0309] 1 H-NMR(400MHz,CDCl3)δ11.94-11.81(m,1H),8.30-8.29(m,1H),7.7-7.36( m,7H),7.03-7.02(m,1H),6.63-6.59(m,1H),5.19(s,2H),5.16-5.14(m,1H ),4.09-3.78(m,2H),3.62-3.46(m,4H),3.38-3.37(m,3H),2.53-2.51(m,1 H),2.37-2.36(m,3H),2.00-1.79(m,2H),1.33(s,12H),1.08-1.05(m,3H).

[0310] Example 27: Preparation of compound (27) (4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl(2-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamido)-4-methoxyphenyl)carbamate

[0311] [ka]

[0312] Step (1): Manufacturing of compound (27)

[0313] In step (2) of Example 25, compound (7) was used instead of compound (1), and the same procedure was repeated to obtain compound (27), the target compound. (Yield 16%)

[0314] 1 H-NMR(400MHz,CDCl3)δ12.30-11.96(m,1H),8.16(s,1H),7.73-7.57(m,6H),7.19-7.05(m,1H),6.82-6.52(m,2H),5.13-5.03(m,3H), 4.01-3.97(m,2H),3.76(s,3H),3.53-3.42(m,4H),2.29(s,3H),2.43-2.34(m,1H),1.94-1.57(m,2H),1.26(s,12H),1.01-0.99(m,3H).

[0315] Example 28: Compound (28) (4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl(6-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidine-3-yl)(methyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide)-1,3-dimethyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazole-5-yl)carbamate:4-(4,4,5,5-tetramethyl l-1,3,2-dioxaborolan-2-yl)benzyl(6-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-7H-p Production of yrrolo[2,3-d]pyrimidine-7-carboxamido)-1,3-dimethyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)carbamate)

[0316] [ka]

[0317] Step (1): Manufacturing of compound (28)

[0318] 133 mg of compound (10) (0.25 mmol, 1 equiv) and 149 mg of 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl carbonochloridate (0.5 mmol, 2 equiv) were dissolved in 4 mL of THF. 85 μL of DIPEA (0.5 mmol, 2 equiv) was added, and the mixture was stirred at 40°C for 16 hours. After the reaction was complete, the mixture was cooled to room temperature and then concentrated under reduced pressure. The concentrate was recrystallized using dichloromethane and ether. The solid obtained by filtration was purified by silica gel column chromatography (dichloromethane:methanol = 20:1) to obtain the target compound (28). (Yield 54%)

[0319] 1H-NMR(300MHz,DMSO-d6)δ12.02-12.02(m,1H),9.37-9.36(m,1H),8.23(s,1H),7.88-7.38 (m,5H),7.16(s,1H),6.91-6.84(m,1H),5.13(s,1H),4.91-4.78(m,2H),4.55-4.54(m,1H) ,4.20-3.72(m,5H),2.73-2.69(m,2H),2.20-2.09(m,3H),1.97-1.85(m,3H),1.74-1.47(m ,3H),1.29-1.23(m,10H),1.16-1.14(m,1H),1.07-1.01(m,3H),0.85-0.81(t,1H,J=6Hz).

[0320] Example 29: Preparation of compound (29)(N-(2-aminophenyl)-8-((3R,4S)-4-ethyl-1-((2,2,2-trifluoroethyl)carbamoyl)pyrrolidin-3-yl)-3H-imidazo[1,2-a]pyrrolo[2,3-e]pyrazine-3-carboxamide)

[0321] [ka]

[0322] Step (1): Production of compound (29-1)

[0323] 200 mg of upadacitinib (0.53 mmol, 1 equiv), 216 mg of bis(4-nitrophenyl)carbonate (0.71 mmol, 1.35 equiv), and DMAP (0.74 mmol, 1.4 equiv) were added to 5 mL of dichloromethane and stirred at room temperature for 1 day. 131 mg of tert-butyl(2-aminophenyl)carbamate (0.63 mmol, 1.2 equiv) dissolved in 3 mL of dichloromethane was added dropwise to the mixture and stirred at room temperature for 16 hours. The concentrate was purified by silica gel column chromatography (dichloromethane:methanol = 10:1) to obtain compound (29-1) of the target compound. (Yield 63%)

[0324] Step (2): Manufacturing of compound (29)

[0325] 203 mg of compound (29-1) (0.33 mmol, 1 equiv) was dissolved in 16 mL of dichloromethane. Then, 4.1 mL of trifluoroacetic acid (20V) was added to the mixture, and the mixture was stirred at room temperature for 16 hours. After the reaction was complete, the reaction solution was cooled to 0-5°C, and neutralization was achieved by adding aqueous NaHCO3 dropwise. Dichloromethane was added to separate the organic layer, which was then dried over magnesium sulfate and filtered. The remaining filtrate was concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (dichloromethane:methanol = 10:1) to obtain the target compound (29). (Yield 81%)

[0326] 1H-NMR(300MHz,DMSO-d6)δ10.76(s,1H),8.88(s,1H),8.09-8.08(d,1H,J=3Hz),7.67(s,1H ),7.46-7.40(m,2H),7.02-6.98(m,2H),6.85-6.83(dd,1H,J=6Hz,0.9Hz),6.67-6.64(td,1 H,J=6Hz,0.9Hz),5.14(s,2H),4.42-4.41(m,1H),3.89-3.80(m,4H),3.72-3.69(m,1H),3.2 7-3.25(m,1H),2.61-2.59(m,1H),1.09-1.05(m,1H),0.82-0.77(m,1H),0.65-0.63(m,3H).

[0327] Example 30: Preparation of compound (30) (N-(2-amino-4-(dimethylamino)phenyl)-8-((3R,4S)-4-ethyl-1-((2,2,2-trifluoroethyl)carbamoyl)pyrrolidin-3-yl)-3H-imidazo[1,2-a]pyrrolo[2,3-e]pyrazine-3-carboxamide)

[0328] [ka]

[0329] Step (1): Production of compound (30-1)

[0330] In step (1) of Example 29, tert-butyl(2-amino-5-(dimethylamino)phenyl)carbamate was used instead of tert-butyl(2-aminophenyl)carbamate, and the same procedure was repeated to obtain compound (30-1) of the target compound. (Yield 52%)

[0331] Step (2): Production of compound (30)

[0332] In step (2) of Example 29, compound (30-1) was used instead of compound (29-1), and the same procedure was repeated to obtain the target compound (30). (Yield 91%)

[0333] 1 H-NMR(300MHz,DMSO-d6)δ10.46(s,1H),8.87(s,1H),8.06-8.06(d,1H,J=1.8Hz),7.66(s,1H),7.38-7.37 (d,1H,J=2.1Hz),7.11-7.10(d,1H,J=4.2Hz),7.00-6.98(t,1H,J=3Hz),6.19-6.19(d,1H,J=1.2Hz),6.08- 6.07(dd,1H,J=3Hz,1.5Hz),4.96(s,2H),4.42-4.41(m,1H),3.91-3.79(m,4H),3.72-3.69(m,1H),3.27-3. 24(m,1H),2.85(s,6H).2.62-2.61(m,1H),1.09-1.03(m,1H),0.82-0.77(m,1H),0.65-0.63(t,3H,J=6Hz).

[0334] Example 31: Compound (31)(N-(6-amino-1,3-dimethyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazole-5-yl)-8-((3R,4S)-4-ethyl-1-((2,2,2-trifluoroethyl)carbamoyl)pyrroridine-3-yl)-3H-imidazo[1,2-a]pyrrolo[2,3-e]pyrazine-3-carboxamide:N-(6-amino-1,3-d Production of imethyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-8-((3R,4S)-4-ethyl-1-((2,2,2-trifluoroethyl)carbamoyl)pyrrolidin-3-yl)-3H-imidazo[1,2-a]pyrrolo[2,3-e]pyrazine-3-carboxamide)

[0335] [ka]

[0336] Step (1): Production of compound (31-1)

[0337] 1.5 g of 5-amino-1,3-dimethyl-6-nitro-1H-benzo[d]imidazol-2(3H)-one (6.75 mmol, 1 equiv) was dissolved in methanol (150 mL), to which 750 mg of 10% Pd / C (10 wt.) was added. The mixture was then stirred under atmospheric pressure and hydrogen gas for 18 hours. The mixture was then heated to room temperature and stirred for 18 hours. After the reaction was complete, the reaction mixture was passed through celite and filtered, and washed with 50 mL of methanol. The filtrate was concentrated under reduced pressure to obtain compound (31-1), the target compound.

[0338] Step (2): Preparation of compound (31-2)

[0339] 1.3 g of compound (31-1) (6.75 mmol, 1 eq.) and (Boc)2 (1.77 g, 8.1 mmol, 1.2 eq.) were dissolved in 100 mL of dichloromethane, and the mixture was cooled to 0°C. 120 mg of NBS (0.675 mmol, 0.1 eq.) was added, and the mixture was heated to room temperature and stirred for 18 hours. After the reaction was complete, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography to obtain (100% ethyl acetate) the target compound (31-2). (2-step yield: 25%)

[0340] Step (3): Production of compound (31-3)

[0341] 570 mg of upadacitinib (1.5 mmol, 1 eq.) and NPC (500 mg, 1.65 mmol, 1.1 eq.) were dissolved in 40 mL of dichloromethane. 0.52 mL of triethylamine (3.75 mmol, 2.5 eq.) was then added to this solution, and the mixture was refluxed and stirred for 18 hours. After the reaction was complete, the reaction mixture was cooled to room temperature, and the target compound (31-3) was obtained without any purification process.

[0342] Step (4): Production of compound (31-4)

[0343] Compound (31-2) (450 mg, 1.5 mmol, 1 eq.) was added to the reaction solution from step (3) above, and the mixture was stirred for 18 hours. After the reaction was complete, the reaction solution was concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography to obtain the target compound (31-4). (Yield 37%)

[0344] Step (5): Manufacturing of compound (31)

[0345] 735 mg of compound (31-4) (1.05 mmol, 1 equiv) was dissolved in a 30 mL solution of dichloromethane, to which 6 mL of TFA was added. The mixture was stirred at room temperature for 2 hours. After the reaction was complete, the reaction solution was concentrated under reduced pressure, and then based by adding an aqueous solution of Na2CO3. Dichloromethane was added for extraction. After separating the organic layer, it was dried over magnesium sulfate and filtered. The remaining filtrate was concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography to obtain the target compound (31). (Yield 85%)

[0346] 1H-NMR(400MHz,DMSO-d6)δ10.75(s,1H),8.89(s,1H),8.09(d,1H,J=4.0Hz),7. 68(s,1H),7.41(d,1H,J=4.0Hz),7.24(s,1H),7.01(t,1H,J=6.0Hz),4.93(s,2H ),4.44-4.40(m,1H),3.90-3.79(m,4H),3.73-3.69(m,1H),3.29-3.25(m,7H), 2.60-2.57(m,1H),1.11-1.05(m,1H),0.85-0.79(m,1H),0.64(t,3H,J=7.2Hz).

[0347] Example 32: Compound (32)(((4-((((6-(8-((3R,4S)-4-ethyl-1-((2,2,2-trifluoroethyl)carbamoyl)pyrroridine-3-yl)-3H-imidazo[1,2-a]pyrrolo[2,3-e]pyrazine-3-carboxamide)-1,3-dimethyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazole-5-yl)carbamoyl)oxy)methyl)phenyl)boric acid:4-((((6-(8-((3R,4 S)-4-ethyl-1-((2,2,2-trifluoroethyl)carbamoyl)pyrrolidin-3-yl)-3H-imidazo[1,2-a]pyrrolo[2,3-e]pyrazine-3 -carboxamido)-1,3-dimethyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)carbamoyl)oxy)methyl)phenyl)boronic production of acid)

[0348] [ka]

[0349] Step (1): Production of compound (32)

[0350] In step (2) of Example 25, compound (31) was used instead of compound (1), and the same procedure was repeated to obtain compound (32), the target compound. (Yield 28%)

[0351] 1 H-NMR(400MHz,DMSO-d6)δ11.57(s,1H),9.44(s,1H),8.59-8.57(m,1H),8.10-8.00(m,2H),7.90-7.69(m,3H),7.51-6.84(m,6H),5.45(s,1 H),4.41-4.34(m,1H),3.88-3.66(m,5H),3.36-3.25(m,9H),2.60-2.5 4(m,1H),1.13-1.05(m,1H),0.84-0.77(m,1H),0.64(t,1H,J=7.2Hz).

[0352] Example 33: Preparation of Compound (33) (2-(9-fluoro-11,17-dihydroxy-10,13,16-trimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthren-17-yl)-2-oxoethyl(2-aminophenyl)carbamate)

[0353] [ka]

[0354] Step (1): Production of compound (33-1)

[0355] 5.0 g of dexamethasone (12.76 mmol, 1 eq.) and pyridine (3.09 mL, 38.27 mmol, 3 eq.) were dissolved in 40 mL of dichloromethane, and the mixture was cooled to 0°C. A 10 mL solution of dichloromethane containing 3.33 g of 4-nitrophenyl carbonochloridate (16.58 mmol, 1.3 eq.) was added dropwise, and the mixture was heated to room temperature and stirred for 1 hour. The mixture was then heated to room temperature and stirred for 18 hours. After the reaction was complete, water was added to the reaction mixture and stirred, and the organic layer was separated. The separated organic layer was washed sequentially with 80 mL of 0.1 M HCl and saturated aqueous solution of NaHCO3, dried over magnesium sulfate, filtered, and the remaining filtrate was concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 1:1) to obtain compound (33-1), the target compound. (Yield 87%)

[0356] Step (2): Manufacturing of compound (33)

[0357] 500 mg (0.9 mmol, 1 Eq) of compound (33-1), 0.59 mL (5.39 mmol, 6 Eq) of NMM, and 291 mg (2.69 mmol, 3 Eq) of o-Phenylenediamine were added to 8 mL of DMF, and the mixture was stirred at room temperature for 3 days. After the reaction was complete, water and ethyl acetate were added to the reaction mixture and extracted. The organic layer was separated, dried over magnesium sulfate, filtered, and the remaining filtrate was concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 3:1) to obtain the target compound (33). (Yield 54%)

[0358] 1H-NMR(400MHz,DMSO-d6)δ8.83(s,1H),7.29(d,1H,J=10.4Hz),7.16(d,1H,J=6.0Hz),6.88(t,1H,J=7.2Hz),6.70( dd,1H,J=7.6,1.2Hz),6.53(t,1H,J=7.2Hz),6.23(d,1H,J=8.0Hz),6.01(s,1H),5.42(d,1H,J=4.0Hz),5.14(s,1H) ,5.04-4.79(m,4H),4.16(br,1H),2.91(br,1H),2.65-2.63(m,1H),2.41-2.31(m,2H),2.20-2.15(m,2H),1.91-1.7 7(m,1H),1.69-1.58(m,2H),1.49(s,3H),1.41-1.36(m,1H),1.11-1.02(m,1H),0.91(s,3H),0.80(d,3H,J=7.2Hz).

[0359] Example 34: Preparation of compound (34) (2-(9-fluoro-11,17-dihydroxy-10,13,16-trimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthren-17-yl)-2-oxoethyl(2-amino-4-methylphenyl)carbamate)

[0360] [ka]

[0361] Step (1): Production of compound (34-2)

[0362] In step (2) of Example 33, compound (34-1) was used instead of o-Phenylenediamine, and the same procedure was repeated to obtain compound (34-2), the target compound. (Yield 30%)

[0363] Step (2): Manufacturing of compound (34)

[0364] In step (2) of Example 1, compound (34-2) was used instead of compound (1-1), and the same procedure was repeated to obtain the target compound (34). (Yield 14%)

[0365] 1 H-NMR(400MHz,DMSO-d6)δ8.75(s,1H),7.29(d,1H,J=10.0Hz),7.02(s,1H,),6.51(s,1H),6.35(d,1H, J=7.6Hz),6.23(d,1H,J=10.0Hz),6.01(s,1H),5.42(d,1H,J=4.4Hz),5.15(s,1H),5.03-4.78(m,4H),4 .16(br,1H),2.91(br,1H),2.67-2.58(m,1H),2.34-2.31(m,2H),2.19-2.12(m,5H),1.91-1.79(m,1H) ,1.68-1.58(m,2H),1.49(s,3H),1.37-1.34(m,1H),1.10-1.07(m,1H),0.90(s,3H),0.81-0.79(m,3H).

[0366] Example 35: Compound (35)((1R,2R,E)-1-((2S,5S,11S,14S,17S,20S,23R,26S,29S,32S)-5-ethyl-11,17,26,29-tetraisobutyl-14,32-diisopropyl-1,7,10,16,20,23,25,28,31-nonamethyl-3,6,9,12,15,18,21,24,27,30,33-undekaoxo-1,4,7,10,13,16,19,22,25,28,31-undekaazacyclotriacontan-2-yl)-2-methylhex-4-en-1-yl(2-aminophenyl)carbamate:(1R,2R,E)-1-( (2S,5S,11S,14S,17S,20S,23R,26S,29S,32S)-5-ethyl-11,17,26,29-tetraisobutyl-14,32-diisopropyl-1,7,10,16,20,23,25,28,31-nonamethyl-3,6,9,12,15, Production of cyclotritriacontan-2-yl)-2-methylhex-4-en-1-yl(2-aminophenyl)carbamate)

[0367] [ka]

[0368] Step (1): Production of compound (35-1)

[0369] To a solution of 120 mg of cyclosporine (0.1 mmol, 1 eq.) dissolved in toluene (15 mL), triphosgene (2.97 g, 10 mmol, 100 eq.) and triethylamine (10 mg, 0.1 mmol, 1 eq.) were added, and the mixture was stirred at 40°C for 16 hours. After the reaction was complete, the reaction mixture was concentrated under reduced pressure to obtain compound (35-1), the target compound.

[0370] Step (2): Manufacturing of compound (35)

[0371] Compound 35-1 (120 mg, 0.1 mmol, 1 eq.), o-Phenylenediamine (108 mg, 1 mmol, 10 eq.), and triethylamine (100 mg, 1.0 mmol, 10 eq.) were added to 5 mL of DMF, and the mixture was stirred at 20°C for 16 hours. After the reaction was complete, the reaction mixture was concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography to obtain the target compound (35). (Yield 10%) Figure 3 shows the NMR data of compound (35) prepared in Example 35.

[0372] MS (m / z): 1337.7 (M+H).

[0373] Comparative example (negative control group): Preparation of compound (36)

[0374] [ka]

[0375] Step (1): Production of compound 36

[0376] In step (1) of Example 1, o-toluidine was used instead of tert-butyl(2-aminophenyl)carbamate, and the same procedure was repeated to obtain the target compound 4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-N-(o-tolyl)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide. (Yield 46%)

[0377] 1H-NMR(300MHz,DMSO-d6)δ11.85-11.83(m,1H),8.43-8.41(d,1H,J=6Hz),8.14-8.05(m,2H),7.78-7.77(m,1H),7.33-7.25(m,2H),7.13-7.08(t, 1H,J=6Hz),6.96-6.92(m,1H),4.92(s,1H),4.20-3.67(m,5H),3.57-3.3 7(m,2H),2.44-2.39(m,4H),1.91-1.58(m,3H),1.04-1.01(d,3H,J=9Hz).

[0378] Experimental Example 1: Confirmation of nitric oxide scavenging ability of nitric oxide-sensitive pharmaceutical compositions (DAF-2 Assay)

[0379] The NO removal ability of nitric oxide-sensitive pharmaceutical compositions containing compounds from Examples 1-22, 29-31, and 33-35 was evaluated by DAF-2 Assay. Using an NO donor SNAP (S-Nitroso-N-Acetyl-D,L-Penicillamine) capable of releasing NO under hydrophilic conditions, the composition was treated and then reacted with DAF-2, which exhibits fluorescence upon binding to NO, to confirm the amount of NO.

[0380] Specifically, to prepare 5 mM DAF-2, 100 mM SNAP, and 5 mM of the compound synthesized in the above example, a reagent stock solution was prepared using DMSO (Dimethyl sulfoxide). For the final concentration for the reaction, 10 μM DAF-2 and 100 μM SNAP were added to a PBS (Phosphate buffered saline) solution to the concentrations shown in Table 1, and the mixture was reacted in an incubator at 37°C for 3 hours. To compare the amount of remaining NO relatively, the NO removal capacity was calculated using Equation 1 below and is shown in Table 10 (N / T: Not Tested).

[0381] [Formula 1] NO removal capacity (%) = 100 - [(Fluorescence value with compound added - background fluorescence value) / (Fluorescence value without any additive - background fluorescence value)] × 100

[0382] [Table 10]

[0383] As a positive control group, we used vitamin B12 (Cyanocobalamin), which is known to have NO scavenging ability.

[0384] Referring to Table 10 above, it was confirmed that the compounds synthesized in the examples had a higher nitric oxide scavenging capacity than vitamin B12 in the positive control group.

[0385] Experimental Example 2: Confirmation of nitric oxide scavenging ability of nitric oxide-sensitive pharmaceutical compositions capable of removing reactive oxygen species (DAF-2 Assay)

[0386] Nitric oxide-sensitive pharmaceutical compositions containing the compounds from Examples 23-28 and 32 can have the ability to remove reactive oxygen species (ROS). The NO removal ability of the pharmaceutical compositions was evaluated by DAF-2 Assay. Using an NO donor SNAP that can release NO under hydrophilic conditions and H2O2 that can release ROS, the composition was treated and then reacted with DAF-2, which exhibits fluorescence upon binding to NO, to confirm the amount of NO.

[0387] Specifically, the compounds synthesized in the above example were added to a 100 μM SNAP solution at the concentrations shown in Table 2 below and reacted. Then, 10 μM DAF-2 was added, and the mixture was reacted in a 37°C incubator for 3 hours. To compare the amount of remaining NO relatively, the NO removal capacity was calculated using Equation 1 above and is shown in Table 11 below.

[0388] Additionally, a stock solution of H2O2 (10M) was added to the reaction solution to achieve a final concentration of SNAP 100 μM and H2O2 1 mM. Then, the compounds synthesized in the above example were added at the concentrations shown in Table 2 below and reacted. The reaction was then carried out in the same manner, and the NO removal capacity was calculated using Equation 1 above and is shown in Table 11 below (N / T: Not Tested).

[0389] [Table 11]

[0390] Referring to Table 11, it was confirmed that the compounds in the above examples contain an active oxygen scavenging agent, and that their nitric oxide scavenging ability is improved when H2O2 is present compared to when it is not present.

[0391] Experimental Example 3: Analysis of nitric oxide sensitivity of nitric oxide-sensitive pharmaceutical compositions (HPLC)

[0392] We confirmed whether a nitric oxide-sensitive pharmaceutical composition containing the compound according to the above example can release a pharmaceutically active low-molecular-weight compound by separating the nitric oxide scavenging agent in the presence of nitric oxide.

[0393] The HPLC analysis sample was prepared as follows: 28 mg of NaNO2 (0.41 mmol, 16.4 equiv) was dissolved in 0.3 mL of water, and the mixture was cooled to 0-5°C. 0.2 mL of sulfuric acid (0.1 mmol, 4 equiv) was added to the mixture, and then 12.2 mg (0.025 mmol, 1 equiv) of compound (4) prepared in Example 4 was dissolved in 0.2 mL of acetone and added. The mixture was stirred at room temperature for 1 hour. After the reaction was complete, water and dichloromethane were added to the reaction mixture for extraction. The organic layer was separated, dried over magnesium sulfate, filtered, and the remaining filtrate was concentrated under reduced pressure.

[0394] The aforementioned analytical samples were subjected to HPLC (Agilent 1200, USA) under the conditions shown in Table 12 below, and the results are shown in Figure 2.

[0395] [Table 12]

[0396] HPLC testing confirmed that the nitric oxide-sensitive pharmaceutical composition allows for the separation of the nitric oxide scavenging agent and the release of a low-molecular-weight compound with pharmaceutical activity in an environment with excess nitric oxide.

[0397] Specifically, referring to Figure 2A, the peak of compound (4) from Example 4, a nitric oxide-sensitive pharmaceutical composition, was detected at 14.2 minutes. In contrast, referring to Figure 2B, the peak of compound (4) disappeared in the environment with excess nitric oxide, and a peak was generated that was detected at 7.7 minutes. Figure 2C shows the results of an HPLC test conducted on tofacitinib, a small molecule compound with pharmaceutically active properties, and a peak was detected at 7.7 minutes. Therefore, Figures 2A to 2C show that compound (4) from Example 4 reacted with excess nitric oxide to release tofacitinib.

[0398] Experimental Example 4: Confirmation of nitric oxide scavenging ability of nitric oxide-sensitive pharmaceutical compositions in immune cells.

[0399] Raw264.7 cell line 8 × 10 4 Cells were spread into 48-well plates at a density of cells / well and cultured at 37°C in DMEM medium containing 10% FBS, 100 U / mL penicillin, and 100 μg / mL streptomycin.

[0400] After culturing the aforementioned medium overnight, the medium was replaced with fresh medium containing 10 μM of a nitric oxide-sensitive pharmaceutical composition containing the compounds of Examples 1-30 and 500 ng / mL of LPS, and the medium was cultured again for 24 hours.

[0401] Finally, to measure the amount of NO produced, Griess assay (Sulfanilamide, N-1-naphthylethylenediamine Dihydrochloride) was used to measure the NO2 present in the cell culture medium. -The amount was measured. Specifically, approximately 100 μL of Griess reagent was mixed with a mixture of 50 μL of cell culture medium and 50 μL of PBS, reacted in a well plate for 10 minutes, and then the absorbance at 540 nm was measured. NO scavenging ability was calculated using Equation 2 below, and the results are shown in Table 13 below (Examples 12 and 13 are based on SD values ​​derived from three or more independent experiments, while the remaining examples are representative values ​​from three or more independent experiments).

[0402] [Formula 2] NO removal capacity (%) = 100 - [(NO value of the group to which the compound of the example was added - NO value of the group with no treatment) / (NO value of the group stimulated with LPS - NO value of the group with no treatment)] × 100

[0403] [Table 13]

[0404] Referring to Table 13 above, it was confirmed that when the compounds according to the above examples were treated, the concentration of nitric oxide decreased even within immune cells.

[0405] Experimental Example 5: Confirmation of the inhibitory effect of nitric oxide-sensitive pharmaceutical compositions on inflammatory cytokine secretion in immune cells.

[0406] RAW264.7 cell line 8 × 10 4 Cells were spread into 48-well plates at a density of cells / well and cultured at 37°C in DMEM medium containing 10% FBS, 100 U / mL penicillin, and 100 μg / mL streptomycin.

[0407] After incubation overnight, the culture medium was replaced with fresh medium containing 10 μM of a nitric oxide-sensitive pharmaceutical composition containing the compounds of Examples 1 and 11-13, and 500 ng / mL of LPS, and the culture was repeated for 24 hours.

[0408] Finally, after obtaining the supernatant, the expression level of IL-6 (Interlukin-6) was analyzed using the Sandwich enzyme-linked immunosorbent assay (ELISA). Specifically, 50 μL of supernatant diluted with standard solution was placed in each well of a well plate coated with an antibody specific to IL-6, and IL-6 was bound to the immobilized antibody. After washing with wash buffer, the mixture was reacted with HRP as a secondary antibody at room temperature for 30 minutes, washed three times with wash buffer, and then colored using TMB (3,3',5,5'-TetraMethyl Benzidine) as a substrate. The reaction was stopped with 1 M sulfuric acid, and the degree of color development was measured by absorbance at 450 nm. Based on the degree of color development, the degree of decrease in IL-6 expression was calculated using Equation 3 below, and is shown in Table 14.

[0409] [Formula 3] IL-6 reduction ability (%) = 100 - [(Quantitative value of IL-6 in the group to which the compound of the example was added - Quantitative value of IL-6 in the group without treatment) / (Quantitative value of IL-6 in the group stimulated with LPS - Quantitative value of IL-6 in the group without treatment)] × 100

[0410] [Table 14]

[0411] Referring to Table 14 above, it was confirmed that when the compounds according to the above examples were treated, the secretion of inflammatory cytokines in immune cells was suppressed.

[0412] Experimental Example 6: Confirmation of nitric oxide scavenging ability, immunotherapy cell proliferation inhibitory effect, and inflammatory cytokine secretion inhibitory effect of nitric oxide-sensitive pharmaceutical compositions.

[0413] Experimental Example 6-1: Confirmation of nitric oxide scavenging ability in spleen cells

[0414] The spleen was isolated from 8-week-old C57BL / 6 mice, and splenocytes were isolated using physical methods.

[0415] The isolated spleen cells were placed in RPMI1640 medium containing 10% FBS, 100 U / mL penicillin, and 100 μg / mL streptomycin in a 5 × 10⁶⁴ 6 After dilution to a cell / mL concentration, 500 μL / well was dispensed into 48-well plates and 100 μL / well into 96-well plates, and the cells were cultured in an incubator for 72 hours. When dispensing spleen cells, an immunoassay was induced by treating with Concanavalin A (5 μg / mL), and at this time, 10 μm of the pharmaceutical composition containing the compound according to the example was added together. After 72 hours, the supernatant was obtained, and the remaining NO level was confirmed by grease analysis. The NO level was calculated using Equation 4 below, and the results are shown in Table 15 below.

[0416] [Formula 4] NO removal capacity (%) = 100 - [(NO value of the group to which the compound of the example was added - NO value of the group with no treatment) / (NO value of the group stimulated with Concanavalin A - NO value of the group with no treatment)] × 100

[0417] Experimental Example 6-2: Confirmation of IFN-γ reduction ability in spleen cells

[0418] NO is an important inflammatory medium and can increase the levels of inflammatory cytokines. Concanavalin A (5 μg / mL), which induces an immune response in spleen cells, and the compounds described in the examples were treated in a concentration-dependent manner to confirm whether the levels of IFN-γ, a representative inflammatory cytokine, decreased. After 72 hours of treatment, the supernatant was collected and analyzed using quantitative enzyme-linked immunosorbent assay (ELISA), and the results are shown in Table 15 below.

[0419] Experimental Example 6-3: Confirmation of the ability of spleen cells to inhibit cell proliferation.

[0420] In Experimental Example 6-1, the cell proliferation ability of spleen cells was confirmed by CCK-8 analysis instead of grease analysis, and the results are shown in Table 15 below.

[0421] [Table 15]

[0422] Referring to Table 15 above, it was confirmed that when the compounds according to the above examples were treated, nitric oxide removal, suppression of inflammatory cytokine secretion, and suppression of cell proliferation occurred in spleen cells.

[0423] Experimental Example 7: JAK1, 2, and 3 Activity Assays of Nitric Oxide-Sensitive Pharmaceutical Compositions

[0424] The activity of the JAK in the compounds used in the examples was confirmed using Eurofins' JAK Human TK Kinase Enzymatic Radiometric Kinase Profiler Assay service.

[0425] After reacting the active sites of JAK1, 2, and 3 with radiation-bound ATP, the activity levels were confirmed. The activity levels of JAK1, 2, and 3 at each concentration are shown in Tables 16 and 17 below. The closer the JAK activity level is to 100, the lower the JAK activity of the compound in the example.

[0426] [Table 16]

[0427] [Table 17]

[0428] Referring to Table 16 above, it became clear that the activity of the nitric oxide-sensitive pharmaceutical composition against JAK1 was significantly lower than that of the positive control group tofacitinib. In particular, as shown in Table 17, it was confirmed that not only against JAK1 but also against JAK2-3, the positive control group tofacitinib showed high activity at 100 nM, while compounds (11)-(13) showed very low activity.

[0429] Experimental Example 8: Dose-increasing experiment of nitric oxide-sensitive pharmaceutical composition

[0430] Compound (1) and tofacitinib from the positive control group were administered orally to experimental animals once daily, with the dose increasing daily, and the maximum tolerable dose in mice was then compared.

[0431] Specifically, the experimental animals were 12 six-week-old male C57BL / 6 mice. The animals were housed in groups of four in polycarbonate cages, and were given free access to food and water. The animal housing room was maintained at a temperature of approximately 20±1°C and a humidity of approximately 50±5%, with a constant 12-hour lighting cycle. After one week of adaptation to the animal housing environment, the animals were randomly divided into three groups of four based on their body weight. The experimental groups and the compounds administered to each group were set as shown in Table 18 below.

[0432] [Table 18]

[0433] The compounds listed in Table 18 were administered orally as a single dose once daily. The dose was increased daily starting with a low dose of 200 mg / kg on day 1, with a common ratio of 2, reaching 400 mg / kg on day 2, 800 mg / kg on day 3, and 1,600 mg / kg on day 4. The presence or absence of death was observed during this period.

[0434] In the group receiving increased doses of compound (1), no deaths occurred within 24 hours after increasing the dose to 1,600 mg / kg. In contrast, in the positive control group receiving tofacitinib, two animals died after receiving 800 mg / kg on day 3 and before the dose on day 4. Therefore, the maximum tolerable dose of tofacitinib in the positive control group was confirmed to be 400 mg / kg, and the maximum tolerable dose of compound (1) in the example was confirmed to be 1,600 mg / kg.

[0435] Experimental Example 9: Evaluation of hematological toxicity after single oral administration of a nitric oxide-sensitive pharmaceutical composition.

[0436] Compounds (1) and (4) were administered orally as a single dose to experimental animals, and their hematological toxicity responses were compared to those of tofacitinib.

[0437] The experimental animals were prepared in the same manner as in the experimental example described above, and the experimental groups and the compounds administered to each experimental group were set as shown in Table 19 below.

[0438] [Table 19]

[0439] The administered compounds listed in Table 19 were given as a single oral dose once daily. Two days after administration, dissection was performed, and whole blood was collected from the heart. To prevent blood coagulation, the blood was placed in an EDTA (ethylene diamine tetraacetic acid) tube, and general blood tests (XN-1000, SYSMEX, Japan) were carried out to evaluate hematological toxicity, which is shown in Table 20 below.

[0440] [Table 20]

[0441] *p<0.05, **p<0.01, and ***p<0.001 are measured by the student t-test compared to the normal group.

[0442] To compare the toxicity of the test substances, hematological toxicity was evaluated after single doses of low concentration (200 mg / kg) and high concentration (800 mg / kg). In all groups administered with compound (1) and compound (4), there was no statistically significant difference compared to the normal control group, and no hematological toxic reactions were observed.

[0443] Statistically significant hematological toxicity, including decreased total white blood cell (WBC), neutrophils (NEU), and lymphocytes (LYM) counts, was observed only in the positive control group that received a high concentration of tofacitinib at 800 mg / kg (p<0.05).

[0444] Therefore, when the same high concentration of 800 mg / kg of the compound was administered orally as a single dose to mice, a comparison of the hematological toxicity results confirmed that the compound in the example had lower hematological toxicity than tofacitinib.

[0445] Experimental Example 10: Efficacy evaluation of nitric oxide-sensitive pharmaceutical compositions for inflammatory bowel disease (DSS-induced ulcerative colitis)

[0446] Experimental Example 10-1: Efficacy Evaluation of Compound (1)

[0447] Compound (1) of the example was administered orally once daily to experimental animals with inflammatory bowel disease, and its disease-relieving efficacy was compared with tofacitinib as a positive control group.

[0448] The experimental animals were prepared in the same manner as in the example above, except that they were randomly divided into four groups of five.

[0449] To establish an inflammatory bowel disease model, 2.5% DSS powder was mixed into drinking water and supplied ad libitum for 5-6 days during a disease induction period, after which normal drinking water was supplied. The experimental groups and the compounds administered to each group were set as shown in Table 21 below.

[0450] [Table 21]

[0451] The administered compounds listed in Table 21 were given orally once daily. Bloody stools, diarrhea, and weight loss were quantified according to the criteria shown in Table 22 below, and the summation was used to formulate the disease activity index (DAI), which is shown in Figure 4.

[0452] [Table 22]

[0453] As shown in Figure 4, statistically significant efficacy was observed for tofacitinib (p<0.01) and compound (1) (p<0.01) on day 7 of the trial. The inflammatory bowel disease relief efficacy of compound (1) was confirmed to be at an average level equivalent to that of tofacitinib.

[0454] Experimental Example 10-2: Efficacy Evaluation of Compound (4)

[0455] In the aforementioned experimental example 10-1, compound (4) was used instead of compound (1), and DSS administration was limited to 5 days, with the drug being administered from the 6th day. The efficacy of compound (4) against inflammatory bowel disease was confirmed in the same manner, as shown in Figure 5.

[0456] As shown in Figure 5, no inflammatory bowel disease relief efficacy of tofacitinib was observed on day 9 of the trial, and statistically significant efficacy was observed only in the compound (4) administration group (p<0.05).

[0457] As seen in Figures 4 and 5, the inflammatory bowel disease alleviation efficacy of compounds (1) and (4) was confirmed to be equivalent to or better than that of tofacitinib.

[0458] Experimental Example 11: Evaluation of the efficacy of nitric oxide-sensitive pharmaceutical compositions for rheumatoid arthritis.

[0459] Experimental Example 11-1: Efficacy Evaluation of Compound (1)

[0460] The disease-relieving efficacy of compound (1) and tofacitinib was compared when administered orally once daily to experimental animals with rheumatoid arthritis.

[0461] The experimental animals consisted of 30 8-week-old DBA / 1 male mice. Each group of six animals was housed in polycarbonate cages, with free access to food and water. The animal housing room was maintained at a temperature of approximately 20±1°C and humidity of approximately 50±5%, with a constant 12-hour lighting cycle. After one week of adaptation to the animal housing environment, the animals were randomly divided into five groups. The normal mouse group consisted of five individuals to minimize the number of animals. The compound (1) administration group consisted of 5-6 mice, the minimum number required to calculate statistics using a restrictive drug dose.

[0462] To establish a rheumatoid arthritis model, a 1:1 mixture of 2 mg / mL chick type II collagen and 1 mg / mL complete freund's adjuvant (CFA) was administered intradermally to the tails of mice at a dose of 100 μL each. Nineteen days later, an emulsion of 2 mg / mL chick type II collagen and incomplete freund's adjuvant (IFA) was administered intradermally to the tails of mice at a dose of 100 μL each. The experimental groups and the administered compounds for each experimental group were set as shown in Table 23 below.

[0463] [Table 23]

[0464] The test substances listed in Table 23 were administered orally daily, depending on the duration and frequency of drug administration. The arthritis score was calculated by assigning numerical values ​​to each of the four limbs according to the criteria shown in Table 24 below, and then summing these scores together to determine the final score, which is shown in Figure 6.

[0465] [Table 24]

[0466] Referring to Figure 6, the tofacitinib administration group and the compound (1) administration group showed similar efficacy in alleviating rheumatoid arthritis symptoms on average from disease induction day 28. From disease induction day 33 (p<0.01) to the end of the experiment, a statistically significant efficacy in alleviating rheumatoid arthritis was observed in the tofacitinib 40 mg / kg administration group. In the compound (1) 30 mg / kg administration group, alleviation was observed from disease induction day 35 (p<0.05), and although no statistically significant alleviation was observed from disease induction day 42 to 47, a statistically significant alleviation was again observed from day 49 to the end of the experiment (p<0.05). The average rheumatoid arthritis score in the compound (1) 30 mg / kg administration group was observed to be at a similar level to the tofacitinib 40 mg / kg administration group, and it was confirmed that the period from disease induction day 42 to 47 where no statistically significant alleviation effect was observed was due to individual differences.

[0467] In the group administered compound (1) at 60 mg / kg, a rheumatoid arthritis-relieving efficacy equivalent to or better than that of the group administered tofacitinib at 40 mg / kg, which is the same dose level as compound (1) at 60 mg / kg, was observed from day 33 of disease induction (p<0.01) until the end of the experiment.

[0468] Therefore, when compound (1) was orally administered once daily at concentrations of 30 mg / kg and 60 mg / kg during the entire experimental period, it was confirmed that it alleviated rheumatoid arthritis at a level equivalent to or better than to tofacitinib at 40 mg / kg.

[0469] Experimental Example 11-2: Efficacy Evaluation of Compound (11)

[0470] The compound (11) described in the example was administered orally once daily to experimental animals with rheumatoid arthritis, and its disease-relieving efficacy was compared with that of tofacitinib, which was used as a positive control group.

[0471] The experimental animals were 31 eight-week-old DBA / 1 male mice. The animals were housed in polycarbonate cages in groups of five, with free access to food and water. The animal housing room was maintained at a temperature of approximately 20±1°C and humidity of approximately 50±5%, with a constant 12-hour lighting cycle. After one week of adaptation to the animal housing environment, the animals were randomly divided into five groups based on body weight. The normal mouse group consisted of five individuals to minimize the number of animals. The compound (11) administration group consisted of five individuals, the minimum number required for statistical analysis using a limiting drug dose. The remaining groups consisted of seven individuals each, with two extra individuals added to account for experimental error.

[0472] To establish a rheumatoid arthritis model, a 1:1 mixture of 2 mg / mL chick type II collagen and 1 mg / mL complete freund's adjuvant (CFA) was intradermally administered to the tails of mice at a dose of 100 μL each. Nineteen days later, an emulsion of 2 mg / mL chick type II collagen and incomplete freund's adjuvant (IFA) was intradermally administered to the tails of mice at a dose of 100 μL each.

[0473] The experimental groups and the compounds administered to each experimental group were set as shown in Table 25 below.

[0474] [Table 25]

[0475] The medication was administered orally daily according to the dosage, frequency, and duration shown in Table 25. The arthritis score for each of the four limbs was quantified according to the criteria shown in Table 24, and the total score was calculated and shown in Figure 7.

[0476] Referring to Figure 7, from day 28 of the trial, the compound (11) administration group and tofacitinib showed similar efficacy in alleviating rheumatoid symptoms on average. Statistically significant efficacy in alleviating rheumatoid arthritis was observed in the tofacitinib administration group on days 39 (p<0.05) and 40 (p<0.01). In the compound (11) administration group, although there was deviation within the group and no statistically significant difference was confirmed, it showed a comparable level of alleviation efficacy to tofacitinib over the entire 40 days.

Claims

1. The compound of the following chemical formula 1, or its solvate, hydrate, stereoisomer, or pharmaceutically acceptable salt. 【Chemistry 1】 (In chemical formula 1, X is a low-molecular-weight compound that contains an amine group or a hydroxyl group in its molecular structure and has pharmaceutically active properties. Y is hydrogen, C １ -C ６ alkyl group, C １ -C ６ Alkoxy groups, hydroxyl groups, nitro groups, amino groups, halogens, thiol groups, cyano groups, and 【Chemistry 2】 It is selected from the group consisting of, Z is, 【Transformation 3】 It is selected from the group consisting of, n and m are independent integers between 0 and 4. R １ and R ２ are each independently, C １ -C ６ alkyl group, C １ -C ６ alkoxy group, hydroxy group, nitro group, amino group, halogen, thiol group, cyano group, C ３ -C １４ aryl group, C ３ -C １４ heteroaryl group, -NR １１ R １２ 、-NR １２ C(=O)R １２ 、-C(=O)R １１ 、and -C(=O)OR １１ and are selected from the group consisting of The aforementioned R １１ and R １２ These are, independently, hydrogen, halogen, and CF ３ , C １ -C ６ alkyl group, C ３ -C １４ Aryl group, and C ３ -C １４ Selected from the group consisting of heteroaryl groups, The aryl group or heteroaryl group can each be independently a halogen, a hydroxyl group, a thiol group, a cyano group, or C1. １ -C ６ Alkyl alkyl groups, and C １ -C ６ Having at least one substituent selected from the group consisting of alkoxy groups, or being unsubstituted, R １ and R ２ Each of them is independent, and if there are two or more, they are either identical or different from each other, and two adjacent R １ or R ２ These may or may not form a ring by arbitrarily joining and fusing. R ３ and R ４ Each of these independently consists of hydrogen, halogen, hydroxyl group, thiol group, cyano group, and C. １ -C ６ Alkyl alkyl groups, and C １ -C ６ Selected from the group consisting of alkoxy groups, R ３ and R ４ (These two elements combine to form a fused ring.)

2. The compound according to claim 1, or a solvate, hydrate, stereoisomer, or pharmaceutically acceptable salt thereof, wherein X is a kinase inhibitor, an anticancer agent, or an anti-inflammatory agent.

3. The kinase inhibitor is selected from the group consisting of Netarsudil, Fostamatinib, Belumosudil, Tofacitinib, Upadacitinib, Baricitinib, Filgotinib, Abrocitinib, Delgocitinib, Oclacitinib, Peficitinib, and Ruxolitinib, and is the compound according to claim 2, or a solvate, hydrate, stereoisomer, or pharmaceutically acceptable salt thereof.

4. The aforementioned anticancer drugs are doxorubicin, cyclophosphamide, cisplatin, oxaliplatin, 5-Fu (5-Fluorouracil), gemcitabine, paclitaxel, docetaxel, irinotecan, and MMAE (Monomethyl auristatin). E) Crizotinib, Osimertinib, Sorafenib, Ibrutinib, Ruxolitinib, Vemurafenib, Ceritinib, Alectinib, Brigatinib, Lorlatinib ), capmatinib, gefitinib, erlotinib, lapatinib, icotinib, afatinib, neratinib, dacomitinib, almonertinib, tucatinib, midostaurine ( Midostaurin, Gilteritinib, Quizartinib, Pexidartinib, Sunitinib, Pazopanib, Vandetanib, Axitinib, Cabozantinib, Regorafenib, A Apatinib, Lenvatinib, Tivozanib, Fluquintinib, Nintedanib, Anlotinib, Erdafitinib, Pemigatinib, Avapritinib, RipretinibPralsetinib, Larotrectinib, Entrectinib, Imatinib, Dasatinib, Nilotinib, Bosutinib, Radotinib, Ponatinib, Acalabrutinib, Zanubrutinib, Fedratinib, Dabrapheni Dabrafenib, Encorafenib, Trametinib, Cobimetinib, Binimetinib, Selumetinib, Palbociclib, Ribociclib, Abemaciclib, Idelalisib, Copanlisib, Duvelisib, Alpelisib pelisib), tazemetostat, vorinostat, belinostat, tucidinostat, panobinostat, enasidenib, ivosidenib, venetoclax, vismodegib, sonidecib, glassdegib, bortezomib (Bort) ezomib, carfilzomib, ixazomib, olaparib, rucaparib, niraparib, talazoparib, umbralisib, trilaciclib, infigratinib, mobocertinib, asciminib, futibatinib,A compound according to claim 2, or a solvate, hydrate, stereoisomer, or pharmaceutically acceptable salt thereof, selected from the group consisting of pacritinib and everolimus.

5. The anti-inflammatory agent is selected from the group consisting of dexamethasone, methotrexate, cyclosporine, acetaminophen, etodolac, piroxicam, and aceclofenac, and is the compound according to claim 2, or a solvate, hydrate, stereoisomer, or pharmaceutically acceptable salt thereof.

6. A compound according to claim 1, or a solvate, hydrate, stereoisomer, or pharmaceutically acceptable salt thereof, selected from the group consisting of the following chemical formulas 2 to 6. 【Chemistry 4】 (In chemical formulas 2 to 6, X, R １ , R ２ , R ３ , R ４ , R １１ , R １２ The definitions of n and m are the same as those for chemical formula 1 above.

7. The compound according to claim 1, or a solvate, hydrate, stereoisomer, or pharmaceutically acceptable salt thereof, selected from the group consisting of the following compounds (1) to (35). 【Transformation 5】 【Transformation 6】 【Transformation 7】 【Transformation 8】 【Chemistry 9】 【Chemistry 10】 【Chemistry 11】 【Chemistry 12】

8. A pharmaceutical composition for the prevention or treatment of diseases involving the generation of nitric oxide, comprising the compound described in claim 1, or a solvate, hydrate, stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

9. The aforementioned disease involving the generation of nitric oxide is Inflammatory diseases selected from the group consisting of inflammatory diseases caused by viral infections, inflammatory diseases caused by bacterial infections, sepsis, and degenerative inflammatory diseases; A neurological disorder selected from the group consisting of Alzheimer's disease, amyotrophic sclerosis, Huntington's disease, Parkinson's disease, and stroke; Cardiovascular disease, such as hypertension or heart failure; Autoimmune diseases selected from the group consisting of rheumatoid arthritis, ulcerative bowel disease, Crohn's disease, systemic lupus erythematosus, psoriasis, multiple sclerosis, and childhood idiopathic arthritis; Allergic diseases selected from the group consisting of asthma, rhinitis, and atopic dermatitis; Cancer selected from the group consisting of lung cancer, breast cancer, colorectal cancer, stomach cancer, liver cancer, brain cancer, pancreatic cancer, thyroid cancer, skin cancer, bone marrow cancer, lymphoma, uterine cancer, cervical cancer, ovarian cancer, kidney cancer, and melanoma; A pharmaceutical composition for the prevention or treatment of a disease involving the generation of nitric oxide, as described in claim 8, which is obesity; myelofibrosis; or hepatic encephalopathy.

10. A pharmaceutical composition for the prevention or treatment of diseases involving the generation of nitric oxide, according to claim 8, which is administered orally, intra-articularly, intravenously, subcutaneously, intramuscularly, intraperitoneally, intranasally, intrapulmonaryly, or rectally.

Images