KR20230066899A - Novel compounds as nlrp3 inhibitor and pharmaceutical composition comprising the same - Google Patents

Abstract

The present invention relates to a compound represented by a chemical formula I as an NLRP3 inhibitor, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition including the same as an active ingredient. The compound, the stereoisomer thereof, or the pharmaceutically acceptable salt thereof, and the pharmaceutical composition including the same as an active ingredient can be advantageously used for preventing or treating NLRP3 activity-related diseases.

Description

Novel compounds as NLRP3 inhibitors and pharmaceutical compositions containing them {NOVEL COMPOUNDS AS NLRP3 INHIBITOR AND PHARMACEUTICAL COMPOSITION COMPRISING THE SAME}

The present invention relates to a compound as an NLRP3 inhibitor, a stereoisomer thereof, a pharmaceutically acceptable salt thereof and a pharmaceutical composition containing the compound, and a method for preventing or treating NLRP3 activity-related diseases using the compound and a use thereof.

NOD-like receptor protein 3 (NLRP3) is a protein-coding gene that contains a nucleotide-binding and oligomerization domain (NOD)-like receptor (NLR). ) belongs to the family of NLRP3 encodes a protein containing a pyrin domain, a nucleotide-binding site domain (NBD) and a leucine-rich repeat (LRR) motif. In response to sterile inflammatory danger signals, NLRP3 interacts with adapter proteins, apoptosis-associated speck-like protein (ASC) and pro-caspase-1 to NLRP3 inflammasome is formed. NLRP3 inflammasome activation can then result in the release of the inflammatory cytokines IL-1β and IL-18, resulting in a number of related diseases.

Active cytokines derived from NLRP3 inflammasome activation are important drivers of inflammation and interact with other cytokine pathways to shape the immune response to infection and injury. For example, IL-1β signaling induces secretion of the proinflammatory cytokines IL-6 and TNF. IL-1β and IL-18 synergize with IL-23 to induce IL-17 production by γδ T cells and memory CD4 Th17 cells in the absence of T cell receptor engagement. IL-18 and IL-12 also synergize to induce IFN-γ production from memory T cells and NK cells that promote Th1 responses.

NLRP3 inflammasome activation is associated with various inflammasome-related diseases/disorders, immune diseases, inflammatory diseases, autoimmune diseases and auto-inflammatory diseases, e.g., auto-inflammatory fever syndromes such as cryopyrin-associated periodic syndromes. syndrome; CAPS) (Mortimer et al., Nature Immunol. 2016, 17(10), 1176-1188); sickle cell disease; systemic lupus erythematosus (SLE); Liver related diseases/disorders such as chronic liver disease, viral hepatitis, non-alcoholic steatohepatitis (NASH, Hugh Thomas Nature Reviews Gastroenterology & Hepatology 2017:14, 197), alcoholic steatohepatitis and alcoholic liver disease (Petrasek et al., J. Clin. Invest.2012, 122, 3476-89; Petrasek et al., Nat. Rev. Gastroenterol. Hepatol.2015, 12, 387-400; Mridha et al., J. Hepatol. 2017, 66, 1037-46); Inflammatory arthritis related disorders such as gout, pseudogout (chondrocalcinosis), osteoarthritis (Ridker et al., N. Engl. J. Med. 2017, 377, 1119-31) and rheumatoid arthritis (Mathews et al., Ann. Rheum. Dis. 2014, 73, 1202-10), acute or chronic arthropathy; Kidney-related diseases such as hyperoxalic aciduria (Knauf et al., Kidney Int. 2013, 84, 895-901), lupus nephritis, hypertensive nephropathy (Krishnan et al., Br. J. Pharmacol. 2016, 173, 752 -65), diabetic nephropathy, also referred to as hemodialysis-related inflammation and diabetic kidney disease, which is a kidney-related complication of diabetes mellitus (type 1, type 2 and diabetes mellitus) (Shahzad et al., Kidney Int. 2015, 87, 74-84]) is known to contribute to the invention and progress.

In addition, the involvement of increased production of IL-1β and IL-18 by the NLRP3 inflammasome is associated with various diseases, such as neuroinflammation-related disorders such as brain infections, acute injuries, multiple sclerosis, Alzheimer's disease and neurodegenerative diseases. (hao et al., Front. Pharmacol. 2015, 6, 262); Cardiovascular/metabolic disorders/diseases such as cardiovascular risk reduction (CvRR), atherosclerosis, type I and type II diabetes and related complications (eg nephropathy, retinopathy), peripheral peripheral artery disease (PAD), acute heart failure and hypertension (Ridker et al., N. Engl. J. Med. 2017, 377, 1119-31); wound healing and scar formation; inflammatory skin diseases such as acne, hidradenitis suppurativa (Sweeney et al., Br. J. Dermatol. 2015, 173, 1361), asthma, sarcoidosis, age-related macular degeneration; Cancer-related diseases/disorders such as myeloproliferative neoplasia, leukemia, myelodysplastic syndrome (MDS), myelofibrosis, lung cancer, colon cancer (Ridker et al., Lancet 2017, 390, 1833-42) It has been shown that it may contribute to onset and progression. Diseases/disorders of an immune or inflammatory nature are generally difficult to diagnose or treat efficiently. Most treatments include treatment of the symptoms, slowing the progression of the disease/disorder, lifestyle changes, and surgery as a last resort (eg, open heart surgery for advanced forms of atherosclerosis). Recent studies have linked mitochondrial dysfunction and NLRP3 activation in neuroinflammation-related diseases such as Parkinson's disease (Sarkar et al.,npj Parkinson's disease 2017, 3:30; Zhou et al.,Nature, 2011, 469, 221). . One of the major problems associated with mitochondrial modulators is poor metabolic stability; Therefore, a selective and stable inhibitor of neuroinflammation of this nature is needed (Lee et al., Eur J. Org. Chem. 2017, 141, 240).

Such inflammasome-related diseases/disorders and others such as autoinflammatory febrile syndrome cryopyrin-related periodic syndrome (eg CAPS), sickle cell disease, chronic liver disease, non-alcoholic steatohepatitis (NASH), gout, Hyperoxaluria, pseudogout (chondrocalcinosis), type I/type II diabetes and related complications (e.g. nephropathy, retinopathy), neuroinflammation-related disorders (e.g. multiple sclerosis, brain infections, acute injury, neurodegenerative disease, Alzheimer's disease), atherosclerosis and cardiovascular risk (eg cardiovascular risk reduction (CvRR), hypertension), hidradenitis suppurativa, wound healing and scar formation, and cancer (eg colon cancer , lung cancer, myeloproliferative neoplasms, leukemia, myelodysplastic syndrome (MDS), myelofibrosis), inhibitors of the NLRP3 inflammasome pathway are needed to provide new and/or alternative therapies.

International Publication Patent WO 2016131098 International Publication Patent WO 2020021447 International Publication Patent WO 2020234715

A critical role for the NLRP3 inflammasome in NASH, Hugh Thomas Nature Reviews Gastroenterology & Hepatology 2017:14, 197 NLRP3 inflammasome blockade reduces liver inflammation and fibrosis in experimental NASH in mice, Mridha, A. R. et al. J. Hepatol. 2017:66, 1037 Mortimer et al., Nature Immunol. 2016, 17(10), 1176-1188 Petrasek et al., J. Clin. Invest.2012, 122, 3476-89 Petrasek et al., Nat. Rev. Gastroenterol. Hepatol. 2015, 12, 387-400 Ridker et al., N. Engl. J. Med. 2017, 377, 1119-31 Mathews et al., Ann. Rheum. Dis.2014, 73, 1202-10 Knauf et al., Kidney Int. 2013, 84, 895-901 Krishnan et al., Br. J. Pharmacol.2016, 173, 752-65 Shahzad et al., Kidney Int. 2015, 87, 74-84 hao et al., Front. Pharmacol.2015, 6, 262 Ridker et al., N. Engl. J. Med. 2017, 377, 1119-31 Ridker et al., Lancet 2017, 390, 1833-42 Sarkar et al.,npj Parkinson's disease 2017, 3:30 Zhou et al., Nature, 2011, 469, 221 Lee et al., Eur J. Org. Chem.2017, 141, 240)

An object of the present invention is to provide a compound represented by Formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an NLRP3 inhibitor.

Another object of the present invention is to provide a pharmaceutical composition comprising a compound as an NLRP3 inhibitor, a stereoisomer thereof or a pharmaceutically acceptable salt thereof.

Another object of the present invention is to provide a composition for preventing or treating NLRP3 activity-related diseases, comprising a compound as an NLRP3 inhibitor, a stereoisomer thereof or a pharmaceutically acceptable salt thereof.

Another object of the present invention is to provide a method for preventing or treating a disease associated with NLRP3 activity, comprising administering the compound in a therapeutically effective amount.

Another object of the present invention is to provide the use of the compound for the prevention or treatment of NLRP3 activity-related diseases or for the preparation of a medicament for the prevention or treatment of NLRP3 activity-related diseases.

Hereinafter, the present invention will be described in more detail. All combinations of the various elements disclosed herein fall within the scope of the present invention. In addition, it cannot be seen that the scope of the present invention is limited by the specific description below.

Compound represented by Formula I

The present invention provides a compound represented by the following formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

<Formula I>

In Formula I,

X ₁ and X ₂ are each independently N or CR ₂ ;

R ₁ is H, C1-C5 alkyl, C1-C5 alkyl-OH, -(C1-C5 alkyl)-O-(C1-C5 alkyl), -(C1-C5 alkyl)-NH ₂ , -(C1-C5 alkyl)-NH-(C1-C5 alkyl), -(C1-C5 alkyl)-N(C1-C5 alkyl) ₂ , cycloalkyl, or heterocycloalkyl;

R ₂ is H, C1-C5 alkyl, C1-C5 alkyl-OH, halogen, CF ₃ , NRaRb or C(=O)O(C1-C5 alkyl);

Ra and Rb are each independently H, C1-C5 alkyl, -C(=O)(C1-C5 alkyl), C1-C5 alkyl-OH, or -S(=O) ₂ (C1-C5 alkyl);

m and n are each independently an integer from 0 to 2;

Z is CH ₂ , NR ₃ or O;

R ₃ is H or C1-C5 alkyl;

W ₁ is O or NRc, Rc is H or CN;

W ₂ is O or S;

는

,

또는

이고,

Is

,

or

ego,

X ₃ is CR ₄ or N, where k is 0 or 1;

R ₄ is H or halogen;

R ₅ and R ₆ are each independently H or C1-C5 alkyl.

In one embodiment, in Formula I, X ₁ and X ₂ are each independently CR ₂ , which means that R ₂ in each of X ₁ and X ₂ may be the same as or different from each other.

In one embodiment, the compound represented by Formula I may be represented by Formula Ia below.

<Formula Ia>

In Formula Ia, each of X ₁ , X ₂ , R ₁ , W ₁ , W ₂ , Z and A is the same as defined in Formula I.

In one embodiment, the compound represented by Formula I may be a compound represented by Formula II below.

<Formula II>

In Formula II above,

X ₁ and X ₂ are each independently N or CR ₂ ;

R ₁ is H, C1-C5 alkyl, C1-C5 alkyl-OH, -(C1-C5 alkyl)-O-(C1-C5 alkyl), -(C1-C5 alkyl)-NH _2, -(C1-C5 alkyl)-NH-(C1-C5 alkyl), -(C1-C5 alkyl)-N(C1-C5 alkyl) ₂ , cycloalkyl, or heterocycloalkyl;

R ₂ is H, C1-C5 alkyl, halogen, CF ₃ , NRaRb or C(=0)O(C1-C5 alkyl);

Ra and Rb are each independently H or C1-C5 alkyl;

Z is NR ₃ or O;

R ₃ is H or C1-C5 alkyl;

는

,

또는

이고,

Is

,

or

ego,

X ₃ is CR ₄ , where k is 0 or 1;

R ₄ is H or halogen;

R ₅ and R ₆ are each independently H or C1-C5 alkyl.

In one embodiment, the compound represented by Formula I may be a compound represented by Formula III below.

<Formula III>

In Formula III,

R ₁ is H or C1-C5 alkyl;

R ₂ is H, C1-C5 alkyl, halogen, CF ₃ , NRaRb or -C(=0)O(C1-C5 alkyl);

Ra and Rb are each independently H or C1-C5 alkyl;

R ₃ is H or C1-C5 alkyl;

는

이며,

Is

is,

X ₃ is CR ₄ , where k is 0 or 1;

R ₄ is H or halogen.

In one embodiment, the compound represented by Formula III is

R ₁ is H or C1-C5 alkyl;

R ₂ is H;

R ₃ is H;

는

이며, R₄는 H 또는 할로겐인 구조를 갖는 것이 바람직하다.

Is

And, R ₄ It is preferable to have a structure of H or halogen.

In one embodiment, the compound represented by Formula I may be a compound represented by Formula IIIa below.

<Formula IIIa>

In the above Formula IIIa,

R ₁ is H or C1-C5 alkyl;

R ₂ is H, C1-C5 alkyl, halogen, CF ₃ , NRaRb or -C(=0)O(C1-C5 alkyl);

Ra and Rb are each independently H or C1-C5 alkyl;

R ₃ is H or C1-C5 alkyl;

는

이며,

Is

is,

X ₃ is CR ₄ , where k is 0 or 1;

R ₄ is H or halogen.

In one embodiment, the compound represented by Formula IIIa,

R ₁ is H or C1-C5 alkyl;

R ₂ is H;

R ₃ is H;

는

이며, R₄는 H 또는 할로겐인 구조를 갖는 것이 바람직하다.

Is

And, R ₄ It is preferable to have a structure of H or halogen.

In one embodiment, the compound represented by Formula I may be a compound represented by Formula IV below.

<Formula IV>

In Formula IV,

R ₁ is H or C1-C5 alkyl;

R ₂ is H, C1-C5 alkyl, halogen, CF ₃ , NRaRb or -C(=0)O(C1-C5 alkyl);

Ra and Rb are each independently H or C1-C5 alkyl;

R ₃ is H or C1-C5 alkyl;

는

이며,

Is

is,

X ₃ is CR ₄ , where k is 0 or 1;

R ₄ is H or halogen.

In the present invention, "Cm-Cn" (where m and n are each independently an integer of 1 or more) means the number of carbon atoms, and for example, 'C1-C5 alkyl' represents alkyl having 1 to 5 carbon atoms.

In the present invention, "alkyl" means a straight-chain or branched-chain saturated hydrocarbon group. In the present invention, alkyl may have 1 to 5 carbon atoms. In one embodiment, an alkyl may have 1 to 3 carbon atoms. Examples of alkyl include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, isobutyl, n-pentyl, sec-pentyl, tert-pentyl, isopentyl, sec-isopentyl, neo-pentyl etc. are mentioned.

In the present invention, "cycloalkyl" means a saturated hydrocarbon ring having 3 or more carbon atoms, and the saturated hydrocarbon ring includes both monocyclic and polycyclic structures. Cycloalkyl can be a saturated hydrocarbon ring having from 3 to 12 carbon atoms. Examples of cycloalkyl include, but are not limited to, one or more selected from cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

In the present invention, "heterocycloalkyl" refers to a cyclic functional group in which at least one carbon atom constituting a ring is substituted with a heteroatom. Examples of the heteroatom may be nitrogen (N), oxygen (O) or sulfur (S). At this time, the heteroatom included in the ring of heterocycloalkyl may be one or two or more, one or one or more heteroatoms may be included, and at least one or more two or more heteroatoms may be included. . Heterocycloalkyls can be 3- to 12-membered rings. Examples of heterocycloalkyl include oxiranyl, oxetanyl, morpholinyl, pyrrolidinyl, piperidinyl, piperazinyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyranyl, tetrahydrothiopyran and the like, but is not limited thereto.

"Halogen" in the present invention may be F, Cl, Br or I.

The compounds according to the present invention, stereoisomers thereof, or pharmaceutically acceptable salts thereof may be compounds listed in Table 1 below.

[Table 1]

In the present invention, "pharmaceutically acceptable salt" means a salt commonly used in the pharmaceutical industry, and can be prepared by a conventional method known to those skilled in the art.

In the present invention, pharmaceutically acceptable salts are, for example, inorganic ion salts prepared with calcium, potassium, sodium or magnesium; inorganic acid salts prepared with hydrochloric acid, nitric acid, phosphoric acid, hydrobromic acid, iodic acid, perchloric acid, or sulfuric acid; Acetic acid, trifluoroacetic acid, citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, manderic acid, propionic acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid organic acid salts prepared from acids, ascorbic acid, carbonic acid, vanillic acid, hydroiodic acid, and the like; sulfonic acid salts made of methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid or naphthalenesulfonic acid, amino acid salts made of glycine, arginine, lysine, etc.; or amine salts made of trimethylamine, triethylamine, ammonia, pyridine, picoline, etc., but the types of salts meant in the present invention are not limited by these listed salts. In one embodiment of the present invention, the salt may be a hydrochloride salt.

In the present invention, "stereoisomer" includes diastereomers and optical isomers, and optical isomers include not only enantiomers but also mixtures and racemates of enantiomers. These isomers can be separated by conventional techniques, e.g., column chromatography or resolution such as HPLC. Alternatively, each stereoisomer of the compound represented by Formula I can be stereospecifically synthesized using optically pure starting materials and/or reagents of known configuration.

In the present invention, "prevention" refers to all activities that suppress or delay the onset of a disease by administering the compound represented by Formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof of the present invention.

In the present invention, "treatment" refers to all activities that improve or beneficially change the symptoms of a suspected or affected subject of a disease by the administration of the compound represented by Formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof of the present invention. .

In the present invention, "NLRP3" refers to nucleic acids, polynucleotides, oligonucleotides, sense and antisense polynucleotide strands, complementary sequences, peptides, polypeptides, proteins, homologous and / or orthologous NLRP3 molecules, isoforms, precursors, mutants, variants , derivatives, splice variants, alleles, other species, and active fragments thereof.

The compound represented by Formula I of the present invention, a stereoisomer thereof or a pharmaceutically acceptable salt thereof may be usefully used for preventing or treating NLRP3 activity-related diseases.

The compound represented by Formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof of the present invention can inhibit NLRP3 or the NLRP3 inflammasome pathway, which induces the production of IL-1β, thereby inhibiting NLRP3 or NLRP3 inflammasome. It may include reducing the capacity of the llamasome pathway.

The compound represented by Formula I of the present invention, a stereoisomer thereof or a pharmaceutically acceptable salt thereof is a drug for the prevention or treatment of diseases related to NLRP3 activity known in the art, similar to or substantially the same level or better than that of NLRP3. It can show the effect of preventing or treating activity-related diseases.

A composition comprising the compound represented by Formula I

The present invention provides a pharmaceutical composition comprising the compound represented by Formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

In addition, the present invention provides a pharmaceutical composition for preventing or treating NLRP3 activity-related diseases, comprising the compound represented by Formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

That is, a pharmaceutical composition comprising the compound represented by Formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient of the present invention can be usefully used for preventing or treating NLRP3 activity-related diseases.

The diseases associated with NLRP3 activity include inflammation, autoimmune diseases, cancer, infections, central nervous system diseases, metabolic diseases, cardiovascular diseases, respiratory diseases, liver diseases, kidney diseases, ocular diseases, skin diseases, lymphatic conditions, psychological disorders, and graft versus host disease, allodynia, wounds, scars, and the like.

Inflammation refers to inflammation that occurs as a result of an inflammatory disorder, such as an autoinflammatory disease, inflammation that occurs as a symptom of a non-inflammatory disorder, inflammation that occurs as a result of infection, or inflammation that occurs as a result of infection or inflammation secondary to injury or autoimmunity. etc. may be included.

Autoimmune diseases include acute disseminated encephalitis, Addison's disease, ankylosing spondylitis, antiphospholipid antibody syndrome (APS), antisynthetase antibody syndrome, aplastic anemia, autoimmune adrenalitis, autoimmune infections, and autoimmune oophoritis. , autoimmune glandular dysfunction, autoimmune thyroiditis, celiac disease, Crohn's disease, type 1 diabetes (T1D), Goodpasture syndrome, Grave's disease, Galambare syndrome (GBS), Hashimoto's disease, idiopathic platelet , reduced purpura, Kawasaki disease, lupus erythematosus, including systemic lupus erythematosus (SLE), primary progressive multiple sclerosis (PPMS), secondary progressive multiple sclerosis (SPMS), and Multiple sclerosis (MS), including relapsing remitting multiple sclerosis (RRMS), myasthenia gravis, opsoclonus myoclonus syndrome (OMS), optic neuritis, Odd's thyroiditis, pemphigus, pernicious anemia, Polyarthritis, primary biliary cirrhosis, rheumatoid arthritis (RA), psoriatic arthritis, juvenile idiopathic arthritis or Still's disease, refractory gouty arthritis, Reiter's syndrome, Sjogren's syndrome, multiple sclerosis systemic connective tissue disorder, Takayasu's arteritis, temporal Arteritis, warm autoimmune hemolytic anemia, Wegener's granulomatosis, alopecia universalis, Behçet's disease, Chagas disease, dysautonomia, endometriosis, hidradenitis suppurativa (HS), interstitial cystitis, neuromuscular dystonia, psoriasis, sarcoidosis idopathy, scleroderma, congestive colitis, Schnitzler syndrome, macrophage activation syndrome, Blau syndrome, vitiligo or vulvar pain, and the like.

Cancers include lung cancer, pancreatic cancer, gastric cancer, myelodysplastic syndrome, leukemia including acute lymphocytic leukemia (ALL) and acute myeloid leukemia (AML), adrenal cancer, anal cancer, basal squamous cell skin cancer , cholangiocarcinoma, bladder cancer, bone cancer, cerebrospinal tumor, breast cancer, cervical cancer, chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), chronic myelomonocytic leukemia (CMML), colorectal cancer, endometrial cancer, esophageal cancer, Ewing's series tumor , including eye cancer, gallbladder cancer, gastrointestinal carcinoid, gastrointestinal stromal tumor (GIST), gestational trophoblastic disease, glioma, Hodgkin's lymphoma, Kaposi's sarcoma, kidney cancer, hypopharyngeal cancer, liver cancer, lung carcinoid, cutaneous T-cell lymphoma lymphoma, malignant mesothelioma, melanoma skin cancer, Merkel cell skin cancer, multiple myeloma, nasal and sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin's lymphoma, non-small cell lung cancer, oral and oropharyngeal cancer, osteosarcoma, ovarian cancer, penile cancer, pituitary tumor, prostate cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, skin cancer, small cell lung cancer, small bowel cancer, soft tissue sarcoma, gastric cancer, testicular cancer, thymic cancer, thyroid cancer including undifferentiated thyroid cancer, uterine sarcoma, vaginal cancer, vulvar cancer, Waldenstrom's macroglobulinemia, Wilms' tumor, and the like.

Infections include viral infections (e.g., influenza virus, human immunodeficiency virus (HIV), alphaviruses (e.g., Chikungunya and Ross River virus), flaviviruses (e.g., dengue virus and Zika virus). viruses), herpes viruses (e.g., Epstein Barr virus, cytomegalovirus, varicella-zoster virus, and KSHV), poxviruses (e.g., vaccinia virus (modified vaccinia virus Ankara) and myxoma virus) , adenovirus (eg adenovirus 5), or papillomavirus), bacterial infection (eg Staphylococcus aureus , Helicobacter pylori ), Bacillus anthracis , Bordatel La Pertussis ( Bordatella pertussis ), Burkholderia pseudomallei ( Burkholderia pseudomallei ), Corynebacterium diphtheriae ( Corynebacterium diptheriae ), Clostridium tetani ( Clostridium tetani ), Clostridium botulinum ( Clostridium botulinum ), Streptococcus pneumoniae Streptococcus pneumoniae , Streptococcus pyogenes, Listeria monocytogenes , Hemophilus influenzae, Pasteurella multicida , Shigella disenteriae ( Shigella dysenteriae ), Mycobacterium tuberculosis ( Mycobacterium tuberculosis ), Mycobacterium leprae ( Mycobacterium leprae ), Mycoplasma pneumoniae ( Mycoplasma pneumoniae ), Mycoplasma hominis ( Mycoplasma hominis ), Neisseria meningitidis ( Neisseria meningitidis ), Neisseria gonorrhoeae ( Neisseria gonorrhoeae ), Rickettsia rickettsii ), Legionella pneumophila ( Legionella pneumophila ), Klebsiella new moniae ( Klebsiella pneumoniae ), Pseudomonas aerugi Nosa ( Pseudomonas aeruginosa ), Propionibacterium acnes ( Propionibacterium acnes ), Treponema pallidum ( Treponema pallidum ), Chlamydia trachomatis ( Chlamydia trachomatis ), Vibrio cholerae ( Vibrio cholerae ), Salmonella typhimurium ( Salmonella typhimurium ), Salmonella typhi , Borrelia burgdorferi or Yersinia pestis , fungal infections (eg from Candida species or Aspergillus species), protozoa infections (e.g. from Plasmodium, Babesia, Giardia, Entamoeba, Leishmania or Trypanosome), helminth infections (e.g. schistosomiasis, roundworms, from tapeworms or flukes), prion infections, and the like.

Central nervous system diseases may include Parkinson's disease, Alzheimer's disease, dementia, motor neuron disease, Huntington's disease, cerebral malaria, brain damage from pneumococcal meningitis, cerebral aneurysm, traumatic brain injury and amyotrophic axial sclerosis, and the like.

Metabolic diseases may include type 2 diabetes (T2D), atherosclerosis, obesity, gout, pseudogout, and the like.

Cardiovascular disease includes hypertension, ischemia, reperfusion injury including ischemic reperfusion injury after MI, stroke including ischemic stroke, transient ischemic attack, myocardial infarction including recurrent myocardial infarction, congestive heart failure and heart failure with preserved ejection fraction. renal failure, embolism, aneurysms, including abdominal aortic aneurysm, or pericarditis, including Dressler syndrome, and the like.

Respiratory diseases may include chronic obstructive pulmonary disorder (COPD), asthma such as allergic asthma and steroid-resistant asthma, asbestosis, silicosis, nanoparticle induced inflammation, cystic fibrosis, idiopathic pulmonary fibrosis, and the like.

Liver disease includes non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH), including advanced fibrosis stage F3 and stage F4, alcoholic fatty liver disease disease: AFLD), and alcoholic steatohepatitis (ASH).

Renal disease may include chronic kidney disease, oxalate nephropathy, nephrocalcinosis, glomerulonephritis, diabetic nephropathy, and the like.

Ocular diseases may include ocular epithelium, age-related macular degeneration (AMD) (dry and wet), uveitis, corneal infection, diabetic retinopathy, optic nerve damage, dry eye, glaucoma, and the like. .

Skin disorders can include dermatitis, such as contact dermatitis and atopic dermatitis, contact hypersensitivity, sunburn, skin lesions, hidradenitis suppurativa (HS), other cyst-causing skin disorders, acne vulgaris, and the like.

Lymphatic conditions may include lymphangitis, Castleman's disease, and the like.

Psychological disorders may include depression, psychological stress, and the like.

The pharmaceutical composition of the present invention may further include one or more pharmaceutically acceptable carriers in addition to the compound represented by Formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof. Pharmaceutically acceptable carriers are commonly used in the art, specifically lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, It may be polyvinylpyrrolidine, cellulose, water, syrup, methyl cellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, minerals, or oil, but is not limited thereto. The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, a dispersing agent, a stabilizer, and the like, in addition to the above components. In addition, the pharmaceutical composition of the present invention can be formulated into oral formulations such as tablets, powders, granules, pills, capsules, suspensions, emulsions, solutions for internal use, emulsions, syrups, external preparations, and suppositories using pharmaceutically acceptable carriers and excipients. Alternatively, it may be formulated in the form of a sterile injectable solution and prepared in a unit dose form, or prepared by placing it in a multi-dose container. The formulation may be prepared by a conventional method used for formulation in the art or a method disclosed in Remington's Pharmaceutical Science (19th ed., 1995), and may be formulated into various formulations according to each disease or component.

Non-limiting examples of preparations for oral administration using the pharmaceutical composition of the present invention include tablets, troches, lozenges, aqueous suspensions, oily suspensions, prepared powders, granules, emulsions, hard capsules, and soft capsules, syrups or elixirs; and the like. In order to formulate the pharmaceutical composition of the present invention for oral administration, a binder such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose or gelatin; excipients such as dicalcium phosphate and the like; disintegrants such as corn starch or sweet potato starch; Lubricants such as magnesium stearate, calcium stearate, sodium stearyl fumarate, or polyethylene glycol wax may be used, and sweeteners, aromatics, syrups, and the like may also be used. Furthermore, in the case of capsules, a liquid carrier such as fatty oil may be additionally used in addition to the above-mentioned materials.

Non-limiting examples of parenteral preparations using the pharmaceutical composition of the present invention include injection solutions, suppositories, powders for respiratory inhalation, aerosols for sprays, ointments, powders for application, oils, creams, and the like. In order to formulate the pharmaceutical composition of the present invention for parenteral administration, sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, external preparations, etc. may be used, and the non-aqueous solvents and suspensions include propylene glycol, polyethylene Glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, etc. may be used, but are not limited thereto.

The present invention provides a method for preventing or treating NLRP3 activity-related diseases, comprising the step of administering the compound represented by Formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof to a subject.

In the present invention, "administration" means introducing a predetermined substance into a subject by an appropriate method.

In the present invention, "subject" refers to all animals such as rats, mice, livestock, etc., including humans who have or may develop NLRP3 activity-related diseases, and may specifically be mammals, including humans, but are not limited thereto no.

The method for preventing or treating NLRP3 activity-related diseases of the present invention may include administering a therapeutically effective amount of the compound represented by Formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

In the present invention, "therapeutically effective amount" means an amount that is sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment and does not cause side effects, which is a patient's sex, age, weight, health Factors including the condition, type of disease, severity, activity of the drug, sensitivity to the drug, method of administration, time of administration, route of administration, rate of excretion, duration of treatment, drugs used in combination or concomitantly, and other factors well known in the medical field can be determined by a person skilled in the art. A specific therapeutically effective amount for a particular patient depends on the type and extent of the response to be achieved, the specific composition, including whether other agents are used as the case may be, the patient's age, weight, general health condition, sex and diet, and the time of administration. , It is preferable to apply differently according to various factors including the route of administration and secretion rate of the composition, treatment period, drugs used together with or concurrently used with the specific composition, and similar factors well known in the medical field.

The present invention provides a use of the compound represented by Formula I, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a composition containing the same for preventing or treating NLRP3 activity-related diseases.

The present invention provides a use of the compound represented by the formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a composition containing the same, for the preparation of a drug for preventing or treating NLRP3 activity-related diseases.

For the preparation of a drug, the compound represented by Formula I, its stereoisomer or its pharmaceutically acceptable salt may be mixed with pharmaceutically acceptable adjuvants, diluents, carriers, etc., and combined with other active agents. It may be manufactured to have a synergistic action.

Matters mentioned in the compounds, pharmaceutical compositions, treatment methods and uses of the present invention apply equally unless contradictory to each other.

a compound represented by formula I of the present invention, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof; And a pharmaceutical composition containing it as an active ingredient can be usefully used for the prevention or treatment of diseases related to NLRP3 activity.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as being limited by these examples.

Preparation of the compound represented by Formula I

The compound represented by Formula I of the present invention can be prepared through the method described below. Unless otherwise stated, starting materials are commercially available or can be prepared by known methods. Any examples provided herein, or use of exemplary language, are merely intended to better illustrate the invention and do not limit the scope of the invention as claimed.

<Example>

Example 1: Synthesis of compound 10, N-((8-fluoro-1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-5,6- Dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(8H)-sulfonamide

[Step 1] Synthesis of Compound A1

Under a nitrogen atmosphere, 1,2,3,5,6,7-hexahydro-s-indacen-4-amine (1 g, 5.7 mmol) was added to tetrahydrofuran (THF, 23 mL) to dissolve, followed by sodium nitrite (418mg, 6.0mmol) was dissolved in water (1mL) and added dropwise at -10°C. The mixture was stirred at -10 °C for 1 hour and then warmed to room temperature (15 to 25 °C). After completion of the reaction, the reaction mixture was washed with water, an aqueous solution of sodium hydrogen carbonate and an aqueous solution of sodium chloride, and the organic layer was extracted with ethyl acetate. The organic layer was dried using sodium sulfate and then concentrated under reduced pressure. The reaction concentrate was purified by silica gel column chromatography (EA (ethyl acetate)/Hexane) to obtain Compound A1 (600 mg, 59%) as a white solid.

¹ H NMR (400 MHz, Chloroform-d) δ 6.86 (s, 1H), 2.87 (s, 8H), 2.10 (quint, J = 7.4, 14.8 Hz, 4H).

[Step 2] Synthesis of Compound A2

Under a nitrogen atmosphere, compound A1 (600mg, 3.4mmol) was added to and dissolved in acetonitrile (ACN, 8.5mL), and then nitrosonium tetrafluoroborate (NOBF ₄ , 501mg, 3.7mmol) was added at 0°C. It was added in portions. The reaction was stirred at 0 °C for 1 hour. After completion of the reaction, the reaction mixture was washed with water and aqueous sodium chloride solution, and the organic layer was extracted with ethyl acetate. The organic layer was dried using sodium sulfate and then concentrated under reduced pressure. The reaction concentrate was purified by silica gel column chromatography (EA/Hexane) to obtain Compound A2 (598mg, 79%) as a yellow solid.

^1H NMR (400MHz, Chloroform-d) δ 5.30 (s, 1H), 3.31 (t, J = 7.4 Hz, 4H), 2.95 (t, J = 7.4 Hz, 4H), 2.18 (quint, J = 7.6, 15.2 Hz, 4H).

[Step 3] Synthesis of Compound A3

Compound A2 (598mg, 2.7mmol) was dissolved in methanol (8.6mL), and then Pd/C (10wt%, 86mg) was added. The reactant was substituted with hydrogen gas several times, and stirred for 16 hours at room temperature under a hydrogen atmosphere. After completion of the reaction, the reaction mixture was filtered using celite and concentrated under reduced pressure. The reaction concentrate was purified by silica gel column chromatography (EA/Hexane) to give Compound A3 (457mg, 88%) as a yellowish white solid.

¹ H NMR (400 MHz, DMSO-d6) δ 4.40 (s, 2H), 2.75 (t, J = 6.6 Hz, 4H), 2.62 (t, J = 6.8 Hz, 4H), 2.50-1.99 (m, 4H) .

[Step 4] Synthesis of Compound A4

Chlorosulfonyl isocyanate (0.03mL, 0.3mmol) was added to isopropyl ether (0.5mL) and dissolved. Here, Compound A3 (70mg, 0.3mmol) dissolved in isopropyl ether (1.5mL) and dichloromethane (0.3mL) was added dropwise at -15°C. The reaction was stirred at -15 °C for 1 hour. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain Compound A4 (100 mg) as a white solid. It was used in the next reaction without further purification.

[Step 5] Synthesis of Compound A5 (Compound 10)

5,6,7,8-tetrahydro-[1,2,4]triazolo-[1,5-a]pyrazine hydrochloride (48mg, 0.4mmol) and pyridine (0.06mL, 0.7mmol) were mixed with dichloromethane ( 2mL), compound A4 (100mg, 0.3mmol) was added. The reaction was stirred at room temperature for 16 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The reaction concentrate was purified by silica gel column chromatography (MC(methylene chloride)/MeOH) to obtain the target compound A5 (Compound 10) (1.8mg, 1%) as a pale yellow solid.

¹ H NMR (400 MHz, DMSO-d6) δ 10.60 (br, 1H), 8.48 (s, 1H), 8.00 (br, 1H), 4.60 (s, 2H), 4.11 (m, 2H), 3.67 (m, 2H), 2.81 (t, J = 6.8 Hz, 4H), 2.63–2.61 (m, 4H), 2.03–1.97 (m, 4H). LC/MS: m/z 421.1 (M+H)+(ES).

Example 2: Synthesis of Compound 1, N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-5,6-dihydroimidazo[ 1,2-a]pyrazine-7(8H)-sulfonamide

[Step 1] Synthesis of Compound B1

Chlorosulfonyl isocyanate (0.25mL, 2.8mmol) was added to isopropyl ether (4mL) and dissolved. Here, 1,2,3,5,6,7-hexahydro-5-indacen-4-amine (500mg, 2.8mmol) dissolved in isopropyl ether (10mL) was added dropwise at -15°C. . The reaction was stirred at -15 °C for 1 hour. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain Compound B1 (688mg, 76%) as a yellow solid.

¹ H NMR (400 MHz, Chloroform-d) δ 7.94 (s, 1H), 7.07 (s, 1H), 2.92-2.81 (m, 8H), 2.13-2.06 (m, 4H).

[Step 2] Synthesis of Compound B2 (Compound 1)

Under a nitrogen atmosphere, compound B1 (30mg, 0.09mmol) and 5,6,7,8-tetrahydroimidazo[1,2- a ]pyrazine (12mg, 0.09mmol) were dissolved in tetrahydrofuran (THF) (1mL). After addition and melting, sodium hydride (60% in mineral oil; 15 mg, 0.3 mmol) was added at -10 °C. The reaction was stirred at -10 °C for 1 hour 30 minutes. Thereafter, the reaction was terminated using an aqueous ammonium chloride solution, and the organic layer was extracted using ethyl acetate. The organic layer was dried using sodium sulfate and then concentrated under reduced pressure. The reaction concentrate was purified by silica gel column chromatography (MC/MeOH) to obtain the target compound B2 (Compound 1) (3.7mg, 10%) as a white solid.

¹ H NMR (400 MHz, MeOH-d4) δ 7.10 (s, 1H), 7.01 (s, 1H), 6.95 (s, 1H), 4.17-4.15 (m, 2H), 3.87-3.84 (m, 2H), 2.84 (t, J = 7.2 Hz, 4H), 2.69 (t, J = 7.2 Hz, 4H), 2.02 (quint, J = 7.6, 7.2 Hz, 4H). LC/MS: m/z 402.1 (M+H)+(ES). HPLC % purity: 94.70%

Example 3: Synthesis of Compound 3, N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-5,6-dihydro-[1 ,2,4]triazolo[1,5-a]pyrazine-7(8H)-sulfonamide

5,6,7,8-tetrahydro-[1,2,4]-triazolo[1,5-a] pyrazine (79 mg, 0.6 mmol) and pyridine (0.10 mL, 1.2 mmol) were mixed with mL) was added and dissolved, and compound B1 (200mg, 0.6mmol) obtained in step 1 of Example 2 was added. Afterwards, the reaction mixture was stirred for 6 hours at room temperature (Room Temperature, RT). After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The reaction concentrate was solidified with dichloromethane and hexane to obtain the target compound D1 (compound 3) (46mg, 18%) as a white solid.

¹ H NMR (400 MHz, DMSO-d6) δ 10.53 (br, 1H), 8.12 (s, 1H), 7.95 (s, 1H), 6.93 (s, 1H), 4.70 (s, 2H), 4.24 (m, 2H), 3.87 (m, 2H), 2.78 (t, J = 7.4 Hz, 4H), 2.53 (t, J = 6.8 Hz, 4H), and 1.95–1.91 (m, 4H). LC/MS: m/z 403.1 (M+H)+(ES).

Examples 4 to 8: Synthesis of compounds 4, 6, 9, 11 and 13

5,6,7,8-tetrahydro-[1,2,4]-triazolo[1,5-a] pyrazine of Example 3, except for using the substituents shown in Table 2, respectively. Each of the compounds of Examples 4 to 8 ( Compounds 4, 6, 9, 11 and 13 ) in Table 3 was synthesized through substantially the same synthesis method as the compound synthesis method of 3.

[Table 2]

[Table 3]

Example 9: Synthesis of compound 2, N-((2,5-diisopropylthiophen-3-yl)carbamoyl)-5,6-dihydroimidazo[1,2-a]pyrazine-7( 8H) -sulfonamide

[Step 1] Synthesis of Compound E1

After dissolving 2,5-dibromothiophene-3-carboxylic acid (4g, 13.9mmol) in tertiary butyl alcohol (40mL), diphenylphosphoryl azide (DPPA) (3mL, 13.9mmol) and N ,N -diisopropylethylamine (DIPEA) (3.7 mL, 20.9 mmol) was added. The reaction was then stirred at 90 °C for 2 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The reaction concentrate was purified by silica gel column chromatography (EA/Hexane) to obtain Compound E1 (3.7 g, 75%) as a pale yellow solid.

¹ H NMR (400 MHz, Chloroform-d) δ7.64 (br, 1H), 6.53 (br, 1H), 1.51 (s, 9H).

[Step 2] Synthesis of Compound E2

After dissolving compound E1 (3.7g, 10.5mmol) in tetrahydrofuran (THF, 74mL) and water (15mL), isopropenyl boronic acid pinacol ester (9.9mL, 52.5mmol), Pd (PPh ₃ ) ₄ (121 mg, 0.1 mmol), and sodium carbonate (3.3 g, 31.5 mmol) were added. The reaction was then stirred at 90° C. for 16 hours. After completion of the reaction, the reaction mixture was washed with water, and the organic layer was extracted with ethyl acetate. The organic layer was dried using sodium sulfate and concentrated under reduced pressure. The reaction concentrate was purified by silica gel column chromatography (EA/Hexane) to give Compound E2 (1.8 g, 63%) as a yellow oil.

¹ H NMR (400 MHz, Chloroform-d) δ 7.54 (br, 1H), 6.70 (br, 1H), 5.33 (s, 1H), 5.30 (s, 1H), 5.17 (s, 1H), 4.92 (s, 1H), 2.12 (s, 3H), 2.09 (s, 3H), 1.50 (s, 9H).

[Step 3] Synthesis of Compound E3

Compound E2 (1.84g, 6.5mmol) was dissolved in methanol (MeOH, 20mL), and then Pd/C (10wt%, 1.0g) was added. The reactants were stirred for 1 hour at a pressure of 40 psi (psi) under a hydrogen atmosphere using a hydrogenation reactor (Pariactor). After completion of the reaction, the reaction mixture was filtered using celite and washed with methanol. The filtered mixture was concentrated under reduced pressure. The reaction concentrate was purified by silica gel column chromatography (EA/Hexane) to obtain Compound E3 (132mg, 7%) as a white solid.

¹ H NMR (400 MHz, Chloroform-d) δ 7.01 (br, 1H), 6.05 (br, 1H), 3.11–3.01 (m, 2H), 1.05 (s, 9H), 1.29–1.24 (m, 12H).

[Step 4] Synthesis of Compound E4

Compound E3 (132mg, 0.4mmol) was added to 4 N HCl (1,4-dioxane solution, 2mL) and then stirred at 0°C for 1 minute. Then, the reaction was stirred at room temperature for 2 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain Compound E4 (48 mg) as an orange solid. It was used in the next reaction without further purification.

¹ H NMR (400 MHz, DMSO-d6) δ 10.14 (br, 2H), 6.71 (s, 1H), 3.40 (quint, J = 6.7, 13.5 Hz, 1H), 3.10 (quint, J = 6.9, 13.7 Hz, 1H), 1.26–1.20 (t, J = 6.6 Hz, 12H).

[Step 5] Synthesis of Compound E5

Chlorosulfonyl isocyanate (48mg, 0.2mmol) was added to and dissolved in diethyl ether (0.2mL). Here, compound E4 (48mg, 0.2mmol) dissolved in diethyl ether (0.8mL) was added dropwise at -15°C. The reaction was then stirred at -15 °C for 20 minutes. After completion of the reaction, the reaction mixture was filtered while washing with diethyl ether to obtain compound E5 (71 mg) as a yellow solid. The obtained compound E5 was used in the next reaction without further purification.

[Step 6] Synthesis of Compound E6 (Compound 2)

Add 5,6,7,8-tetrahydroimidazo[1,2 - a]pyrazine (27 mg, 0.22 mmol) and triethylamine (TEA, 0.10 mL, 0.65 mmol) to dichloromethane (DCM, 2 mL). After melting, compound E5 (71mg, 0.22mmol) was added. Then, the reaction was stirred at room temperature for 16 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The reaction concentrate was purified by silica gel column chromatography (MC/MeOH) to obtain the target compound E6 (Compound 2) (20mg, 22%) as a beige solid.

¹ H NMR (400 MHz, DMSO-d6) δ 8.06 (s, 1H), 7.15 (s, 1H), 6.92 (s, 1H), 6.85 (s, 1H), 4.05 (s, 2H), 4.07 (m, 2H), 3.73 (m, 2H), 3.09–2.99 (m, 2H), 1.20 (d, J = 7.2 Hz, 6H), 1.15 (d, J = 6.8 Hz, 6H). LC/MS: m/z 412.1 (M+H) ⁺ (ES).

Example 10: Synthesis of compound 5, N-((2,6-diisopropylphenyl)carbamoyl)-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-sulfonamide

[Step 1] Synthesis of Compound F1

Chlorosulfonyl isocyanate (0.15mL, 1.6mmol) was added to and dissolved in diethyl ether (2.0mL). Here, 2,6-diisopropylaniline (300mg, 1.6mmol) dissolved in diethyl ether (6.2mL) was added dropwise at -15°C. The reaction was then stirred at -15 °C for 1 hour. After completion of the reaction, the reaction mixture was filtered while washing with diethyl ether to obtain Compound F1 (150 mg) as a white solid. It was used in the next reaction without further purification.

[Step 2] Synthesis of Compound F2 (Compound 5)

After dissolving 5H,6H,7H,8H-imidazo[1,2-a]pyrazine (58mg, 0.4mmol) and pyridine (0.06mL, 0.71mmol) in dichloromethane (DCM) (1.0mL), Compound F1 (150mg, 0.4mmol) was added in several portions at 0°C. Then, the reaction was stirred at room temperature for 1 hour. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The reaction concentrate was purified by silica gel column chromatography (MC/MeOH) to obtain the target compound F2 (Compound 5) (29mg, 15%) as a white solid.

¹ H NMR (400 MHz, DMSO-d6) δ 10.73 (br, 1H), 7.90 (s, 1H), 7.25 (t, J = 7.6 Hz, 1H), 7.13 (d, J = 7.6 Hz, 3H), 6.89 (s, 1H), 4.46 (s, 2H), 4.06 (m, 2H), 3.70 (m, 2H), 3.01–2.96 (m, 2H). 1.06 (s, 12H). LC/MS: m/z 406.2 (M+H)+(ES).

Example 11: Synthesis of Compound 7, 1,2,3,5,6,7-hexahydro-s-indacen-4-yl((5,6-dihydroimidazo[1,2-a]pyrazine -7(8H)-yl)sulfonyl)carbamate

[Step 1] Synthesis of Compound G1

After dissolving 4-bromo-1,2,3,5,6,7-hexahydro-s-indacene (1.56g, 6.58mmol) in tetrahydrofuran (50mL), n-butyllithium ( n -BuLi) (2.5M hexane solution, 0.26mL, 9.87mmol) was added dropwise at -78°C. The reaction mixture was stirred at -78 °C for 30 minutes, then trimethylborate was added. After the reaction mixture was stirred at room temperature for 30 minutes, acetic acid (AcOH) (0.60mL) and hydrogen peroxide (0.41mL) were added and stirred for an additional 30 minutes. Thereafter, the reaction was terminated with aqueous ammonium chloride solution, and the organic layer was extracted using dichloromethane. The organic layer was dried over sodium sulfate and concentrated under reduced pressure to obtain Compound G1 (200 mg) as a white solid. It was used in the next reaction without further purification.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.43 (s, 1H), 6.54 (s, 1H), 2.72 (p, J = 8.4 Hz, 7.2 Hz, 8H), 1.95 (p, J = 7.2 Hz, 6.8Hz, 4H)

[Step 2] Synthesis of Compound G2

Chlorosulfonyl isocyanate (0.10mL, 1.15mmol) was added to isopropyl ether (3mL) and dissolved. To this, compound G1 (200mg, 1.15mmol) dissolved in isopropyl ether (5mL) was added at -15°C. The reaction was then stirred at -15 °C for 1 hour. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain Compound G2 (270mg, 74.5%) as a pale yellow solid. It was used in the next reaction without further purification.

[Step 3] Synthesis of Compound G3 (Compound 7)

Compound G2 (180mg, 0.57mmol) and 5,6,7,8-tetrahydroimidazo[1,2- a ]pyrazine (69mg, 0.57mmol) were dissolved in dichloromethane (5mL) and dissolved in triethylamine. (0.24mL, 1.71mmol) was added at room temperature. Then, the reaction was stirred at room temperature for 3 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The reaction concentrate was purified by silica gel column chromatography (MC/MeOH) to obtain the target compound G3 (Compound 7) (6mg, 2.62%) as a white solid.

^1H NMR (400MHz, DMSO-d6) δ 7.40 (s, 1H), 7.31 (s, 1H), 6.85 (s, 1H), 4.59 (s, 1H), 4.9 (t, 2H), 3.66 (t , J = 5.2 Hz, 2H), 2.78 (t, J = 7.2 Hz, 4H), 2.56 (t, J = 6.8 Hz, 4H), 1.95 (p, J = 7.6 Hz, 7.2 Hz, 4H). LC/MS: m/z 403 (M+H)+(ES).

Example 12: Synthesis of Compound 8, N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)(methyl)carbamoyl)-5,6-dihydro Imidazo[1,2-a]pyrazine-7(8H)-sulfonamide

[Step 1] Synthesis of Compound H1

After dissolving 1,2,3,5,6,7-hexahydro-s-indacen-4-amine (1.00g, 5.8mmol) in 1,4-dioxane (30mL), Cu(OAc) ₂ (2.62g, 14.4mmol), methyl boronic acid (380mg, 6.34mmol), and pyridine (0.93mL, 11.5mmol) were added at room temperature. Then, the reaction was stirred at 100 °C for 16 hours. After completion of the reaction, the reaction mixture was filtered using celite and washed with dichloromethane. The filtered mixture was concentrated under reduced pressure. The reaction concentrate was purified by silica gel column chromatography (EA/Hexane) to obtain Compound H1 (636mg, 58.9%) as a light brown solid.

¹ H NMR (400 MHz, DMSO-d6) δ 6.41 (s, 1H), 4.51 (s, 1H), 2.86 (s, 3H), 2.77–2.67 (m, 8H), 1.99–1.90 (m, 4H). LC/MS: m/z 188 (M+H)+(ES).

[Step 2] Synthesis of compound H2

Chlorosulfonyl isocyanate (0.09mL, 1.07mmol) was added to isopropyl ether (3mL) and dissolved. Here, Compound H1 (200mg, 1.15mmol) dissolved in isopropyl ether (5mL) was added dropwise at -15°C. The reaction was then stirred at -15 °C for 1 hour. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain Compound H2 (170 mg) as a pale yellow solid. It was used in the next reaction without further purification.

[Step 3] Synthesis of Compound H3 (Compound 8)

After dissolving compound H2 (170mg, 0.52mmol) and 5,6,7,8-tetrahydroimidazo[1,2- a ]pyrazine (64mg, 0.52mmol) in dichloromethane (5mL), triethyl Amine (0.22mL, 1.55mmol) was added at room temperature. Then, the reaction was stirred at room temperature for 3 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The reaction concentrate was purified by silica gel column chromatography (MC/MeOH) to obtain the target compound H3 (Compound 8) (3mg, 1.40%) as a light brown solid.

¹ H NMR (400 MHz, DMSO-d6) δ 7.16 (s, 1H), 7.08 (s, 1H), 6.96 (s, 1H), 4.44 (s, 2H), 4.04 (s, 2H), 3.67 (s, 2H), 2.94 (s, 3H), 2.85–2.64 (m, 8H), 2.03–1.95 (m, 4H). LC/MS: m/z 416 (M+H)+(ES).

Examples 13 and 14: Synthesis of compounds 12 and 19, N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-8-methyl-5 ,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazine-7(8H)-sulfonamide and hydrochloride thereof

[Step 1] Synthesis of Compound I1

After dissolving 2-amino-3-methylpyrazine (300mg, 2.75mmol) in toluene (5mL), N,N -dimethylformamide dimethyl acetal (360mg, 3.02mmol) was added at room temperature. Then, the reaction was stirred at 100 °C for 3 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain Compound I1 (451 mg) as a yellow oil. It was used in the next reaction without further purification.

[Step 2] Synthesis of Compound I2

Compound I1 (451mg, 2.75mmol) and sodium acetate (248mg, 3.0mmol) were added and dissolved in methanol (5mL). Here, hydroxylamine hydrochloride (248mg, 3.0mmol) dissolved in methanol (3mL) was added at 0°C. Then, the reaction was stirred at 0 °C for 4 hours. After completion of the reaction, the produced solid was washed with 7M aqueous ammonia dissolved in dichloromethane and methanol and filtered. Compound I2 (418mg, 99%) was obtained as a white solid through a filter. It was used in the next reaction without further purification.

¹ H NMR (400 MHz, DMSO-d6) δ 8.73 (s, 1H), 8.70 (s, 1H), 8.57 (s, 1H), 3.15 (s, 3H). LC/MS: m/z 153 (M+H)+(ES).

[Step 3] Synthesis of Compound I3

Compound I2 (418mg, 2.75mmol) was added to polyphosphoric acid (5mL) and stirred at 90°C for 4 hours. After completion of the reaction, the reaction mass was cooled to 0 °C. The reaction was terminated by adjusting the pH to 9 to 10 by adding an aqueous solution of sodium hydrogen carbonate to the reactant. Then, the organic layer was extracted using dichloromethane. The organic layer was dried using sodium sulfate and concentrated under reduced pressure to obtain Compound I3 (174mg, 47%) as a white solid.

¹ H NMR (400 MHz, Chloroform-d) δ 8.44 (m, 2H), 8.06 (d, J = 4.4 Hz, 1H), 2.96 (s, 3H). LC/MS: m/z 135 (M+H)+(ES).

[Step 4] Synthesis of Compound I4

Compound I3 (174mg, 1.29mmol) was dissolved in methanol (10mL), and then Pd/C (10wt%, 17mg) was added. The reactants were substituted with hydrogen gas several times, and stirred for 16 hours at room temperature under a hydrogen atmosphere. After completion of the reaction, the reactant was filtered using celite and washed with methanol. The filtered mixture was concentrated under reduced pressure. The reaction concentrate was purified by silica gel column chromatography (MC/MeOH) to give Compound I4 (179mg, 99%) as a yellow oil.

¹ H NMR (400 MHz, DMSO-d6) δ 7.88 (s, 1H), 4.05–3.96 (m, 3H), 3.17–2.69 (m, 2H), 1.36 (d, J = 7.2 Hz, 3H). LC/MS: m/z 138 (M+H)+(ES).

[Step 5] Synthesis of Compound I5 (Compound 12)

After dissolving compound I4 (174mg, 1.29mmol) and compound B1 (338mg, 1.07mmol) obtained in step 1 of Example 2 in dichloromethane (5mL), pyridine (0.18mL, 2.15mmol) was added at room temperature. did Then, the reaction was stirred at room temperature for 4 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The reaction concentrate was purified by silica gel column chromatography (MC/MeOH) to obtain the target compound I5 (Compound 12) (10mg, 2%) as a white solid.

¹ H NMR (400 MHz, DMSO-d6) δ 10.52 (br, 1H), 8.10 (s, 1H), 7.92 (s, 1H), 6.88 (s, 1H), 5.35-5.33 (m, 2H), 4.23- 4.09 (m, 3H), 3.80–3.73 (m, 1H), 2.76 (t, J = 7.2 Hz, 4H), 2.47–2.40 (m, 4H), 1.90 (t, J = 7.2 Hz, 4H), 1.51 (d, J = 6.8 Hz, 3H). LC/MS: m/z 417 (M+H)+(ES).

[Step 6] Synthesis of Compound I6 (Compound 19)

Compound I5 (5mg, 0.01mmol) obtained in step 5 was added to 4 N HCl (1,4-dioxane solution, 2mL), followed by stirring at 0°C for 1 minute. Then, the reaction was stirred at room temperature for 10 minutes. After completion of the reaction, the produced solid was filtered and washed with 1,4-dioxane. The target compound I6 (compound 19, hydrochloride salt of compound 12) (48 mg) was obtained as a white solid under reduced pressure.

¹ H NMR (400 MHz, DMSO-d6) δ 10.69 (s, 1H), 8.40 (s, 1H), 7.94 (s, 1H), 6.89 (s, 1H), 5.36-5.31 (m, 1H), 4.23- 4.07 (m, 3H), 3.80-3.73 (m, 1H), 2.74 (t, J = 7.0 Hz, 4H), 2.45-2.36 (m, 4H), 1.89 (t, J = 7.2 Hz, 4H), 1.50 (d, J = 6.8 Hz, 3H). LC/MS: m/z=417 (M+H)+(ES).

Example 15: Synthesis of Compound 17, N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-5-methyl-5,6-di Hydro-[1,2,4]triazolo[1,5-a]pyrazine-7(8H)-sulfonamide

Except for using 2-amino- 6 -methylpyrazine instead of 2-amino-3-methylpyrazine in step 1 of Example 13, the synthesis method substantially the same as the synthesis method for preparing compound I5 in Example 13 Through this, the target compound 17 (15mg, 1.6%) as a beige solid was synthesized.

¹ H NMR (400 MHz, DMSO-d6) δ 7.93 (s, 1H), 7.84 (s, 1H), 6.85 (s, 1H), 4.57-4.40 (m, 3H), 3.81 (d, J = 10 Hz, 1H), 3.60 (m, 1H), 2.77 (t, J = 7.2 Hz, 4H), 2.62 (t, J = 7.2 Hz, 4H), 1.92 (t, J = 7.2 Hz, 4H), 1.42 (d, J = 6.4 Hz, 3H). LC/MS: m/z=417 (M+H)+(ES).

Examples 16 and 17: Synthesis of compounds 14 and 15, 2-(dimethylamino)-N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carba Moyl)-5,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazine-7(8H)-sulfonamide and N-((1,2,3,5,6, 7-hexahydro-s-indacen-4-yl)carbamoyl)-2-(methylamino)-5,6-dihydro-[1,2,4]triazolo[1,5-a] pyrazine- 7(8H)-sulfonamide

[Step 1] Synthesis of Compound J1

After adding and dissolving 2-aminopyrazine (1.0g, 10.5mmol) in 1,4-dioxane (12mL), ethoxycarbonyl isothiocyanate (1.49mL, 12.6mmol) was added dropwise at 0°C. did Thereafter, the reaction mixture was stirred at room temperature for 16 hours, and the reaction mixture was concentrated under reduced pressure. After dissolving methanol (8mL) and ethanol (8mL) in the reaction concentrate, triethylamine (4.4mL, 31.6mmol) and hydroxylamine hydrochloride (2.9g, 42.1mmol) were added. The reaction was stirred for 4 hours under reflux conditions (80-90 °C). After completion of the reaction, the mixture was cooled to room temperature and concentrated under reduced pressure. The reaction concentrate was recrystallized from ethanol to obtain light brown compound J1 (1.2 g, 70%).

[Step 2] Synthesis of compounds J2 and J3

Compound J1 (500mg, 3.70mmol) was added and dissolved in tetrahydrofuran (15mL), and then sodium hydride (296mg, 7.4mmol) was added at 0°C. After the reaction was stirred at 0° C. for 15 min, iodomethane (1.4 mL, 22.2 mmol) was added. Then, the reaction was stirred at room temperature for 16 hours. The reaction mixture was terminated with water, and the organic layer was extracted with ethyl acetate. The organic layer was dried using magnesium sulfate and concentrated under reduced pressure. The reaction concentrate was purified by silica gel column chromatography (MC/MeOH) to obtain Compound J2 (70mg, 12%) and Compound J3 (120 mg, 22%) as white solids.

Compound J2: ^1H NMR (400MHz, Chloroform-d) δ 8.87 (s, 1H), 8.28 (d, J = 2.4 Hz, 1H), 7.98 (d, J = 4.4 Hz, 1H), 3.18 (s, 6H) ). LC/MS: m/z 164.1 (M+H)+(ES).

Compound J3: ^1H NMR (400MHz, Chloroform-d) δ 8.88 (s, 1H), 8.29 (d, J = 3.6 Hz, 1H), 8.00 (d, J = 3.6 Hz, 1H), 4.68 (br, 1H) ) 3.10 (d, J = 4.0 Hz, 3H). LC/MS: m/z 149.1 (M+H)+(ES).

[Step 3-1] Synthesis of Compound J4

After compound J2 (70mg, 0.43mmol) was added to and dissolved in ethanol (2.2mL), platinum dioxide (9.7mg, 0.043mmol) was added. The reactant was substituted with hydrogen gas several times, and stirred for 24 hours at room temperature under a hydrogen atmosphere. After completion of the reaction, the reaction mixture was filtered using celite and washed with ethyl acetate. The filtered mixture was concentrated under reduced pressure. The reaction concentrate was purified by silica gel column chromatography (MC/MeOH) to obtain Compound J4 (61mg, 85%) as a white solid.

¹ H NMR (400 MHz, Chloroform-d) δ 4.02 (s, 2H), 3.97 (t, J = 5.2 Hz, 2H), 3.27 (t, J = 5.4 Hz, 2H), 2.97 (s, 6H). LC/MS: m/z 168.1 (M+H)+(ES).

[Step 4-1] Synthesis of Compound J5 (Compound 14)

Compound J4 (53mg, 0.32mmol) and pyridine (0.051mL, 0.64mmol) were added and dissolved in dichloromethane (1mL), and compound B1 (100mg, 0.32mmol) was added at 0°C. Then, the reaction was stirred at room temperature for 4 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The reaction concentrate was solidified with dichloromethane and hexane to obtain the target compound J5 (Compound 14) (1.8mg, 1%) as a white solid.

¹ H NMR (400 MHz, MeOH-d4) δ 6.95 (s, 1H), 4.66 (s, 2H), 4.11 (t, J = 5.2 Hz, 2H), 3.93 (t, J = 5.6 Hz, 2H), 2.94 (s, 6H), 2.84 (t, J = 7.2 Hz, 4H), 2.67 (t, J = 7.0 Hz, 4H), 2.03 (quint, J = 14.5, 7.3 Hz, 4H). LC/MS: m/z 446.1 (M+H)+(ES).

[Step 3-2] Synthesis of Compound J6

Compound J3 (60mg, 0.40mmol) was dissolved in ethyl acetate (2mL) and methanol (2mL), and then platinum dioxide (9.1mg, 0.04mmol) was added. The reactants were substituted with hydrogen gas several times, and stirred for 24 hours at room temperature under a hydrogen atmosphere. After completion of the reaction, the reaction mixture was filtered using celite and washed with ethyl acetate. The filtered mixture was concentrated under reduced pressure to obtain Compound J6 (59mg, 96%) as a white solid.

¹ H NMR (400 MHz, Chloroform-d) δ 4.05–3.97 (m, 5H), 3.29 (t, J = 5.4 Hz, 2H), 2.92 (d, J = 5.2 Hz, 3H). LC/MS: m/z 154.1 (M+H)+(ES).

[Step 4-2] Synthesis of Compound J7 (Compound 15)

Compound J6 (49mg, 0.32mmol) and pyridine (0.051mL, 0.64mmol) were added and dissolved in dichloromethane (1mL), and compound B1 (100mg, 0.32mmol) was added at 0°C. Then, the reaction was stirred at room temperature for 4 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The reaction concentrate was purified by silica gel column chromatography (MC/MeOH) and solidified with dichloromethane and hexane to obtain the target compound J7 (Compound 15) (7.7 mg, 7%) as an ivory solid.

¹ H NMR (400 MHz, DMSO-d6) δ 10.51 (br, 1H), 8.09 (s, 1H), 6.93 (s, 1H), 5.76 (q, J = 4.4 Hz, 1H), 4.51 (s, 2H) , 3.99 (t, J = 5.2 Hz, 2H), 3.78 (t, J = 5.2 Hz, 2H), 2.78 (t, J = 7.4 Hz, 4H), 2.66 (d, J = 5.2 Hz, 3H), 2.58 (t, J = 7.2 Hz, 4H), 1.94 (quint, J = 14.7, 7.4 Hz, 4H). LC/MS: m/z 432.1 (M+H)+(ES).

Example 18: Synthesis of compound 18, 2-amino-N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-5,6-di Hydro-[1,2,4]triazolo[1,5-a]pyrazine-7(8H)-sulfonamide

[Step 1] Synthesis of Compound K1

After dissolving compound J1 (100mg, 0.74mmol) and di-tert-butyl dicarbonate (194mg, 0.89mmol) obtained in step 1 of Example 16 in tetrahydrofuran (THF, 3mL), lithium bis(trimethyl Silyl)amide (1.0M tetrahydrofuran solution, 1.85mL, 1.85mmol) was added at -78°C. Then, it was stirred for 2 hours at -78°C. Then, the reaction was terminated using an aqueous ammonium chloride solution, and the organic layer was extracted using dichloromethane. The organic layer was dried using magnesium sulfate, and then concentrated under reduced pressure. The reaction concentrate was purified by silica gel column chromatography (MC/MeOH) to obtain the target compound K1 (100mg, 58%) as an ivory solid.

¹ H NMR (400 MHz, DMSO-d6) δ 10.54 (br, 1H), 9.17 (s, 1H), 8.96-8.94 (m, 1H), 8.20-8.18 (m, 1H), 1.49 (s, 9H). LC/MS: m/z 236.0 (M+H)+(ES).

[Step 2] Synthesis of Compound K2

Compound K1 (100mg, 0.43mmol) was dissolved in a 1:1 (v/v) mixed solution (4.3mL) of ethyl acetate and methanol, and then platinum dioxide (10mg, 0.043mmol) was added. The reactant was stirred for 24 hours at room temperature under a hydrogen gas atmosphere. After completion of the reaction, the reactant was filtered through a celite pad and concentrated under reduced pressure to obtain compound K2 (89mg, 88%) as a white solid.

¹ H NMR (400 MHz, chloroform-d) δ 7.78 (br, 1H), 4.12–4.09 (m, 4H), 3.30 (t, J = 5.2 Hz, 2H), 1.52 (s, 9H). LC/MS: m/z 240.0 (M+H)+(ES).

[Step 3] Synthesis of Compound K3

After dissolving compound K2 (89mg, 0.37mmol) and pyridine (0.06mL, 0.74mmol) in dichloromethane (1mL), compound B1 (117mg, 0.37mmol) obtained in step 1 of Example 2 was dissolved at 0°C. added. Then, the reaction was stirred at room temperature for 4 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The reaction concentrate was purified by silica gel column chromatography (MC/MeOH), and solidified with dichloromethane and hexane to obtain compound K3 (31.6mg, 16%) as an ivory solid.

¹ H NMR (400 MHz, DMSO-d6) δ 10.55 (br, 1H), 9.64 (s, 1H), 8.12 (s, 1H), 6.93 (s, 1H), 4.61 (s, 2H), 4.12 (m, 2H), 3.85 (m, 2H), 2.77 (t, J = 7.0 Hz, 4H), 2.55 (t, J = 7.0 Hz, 4H), 1.99–1.92 (m, 4H), 1.43 (s, 9H). LC/MS: m/z 518.1 (M+H)+(ES).

[Step 4] Synthesis of Compound K4 (Compound 18)

After compound K3 (13.9mg, 0.027mmol) was added and dissolved in dichloromethane (0.45mL), trifluoroacetic acid (0.15mL) was added at 0°C. Then, the reaction was stirred at room temperature for 2 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, purified by silica gel column chromatography (MC/MeOH), and solidified with dichloromethane and hexane to obtain the target compound K4 (Compound 18) (1.23mg, 11%) as a white solid. obtained.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 10.53 (br, 1H), 8.12 (s, 1H), 6.93 (s, 1H), 5.30 (br, 2H), 4.38 (s, 2H), 3.95 (m , 2H), 3.76 (m, 2H), 2.78 (t, J = 7.2 Hz, 4H), 2.59 (t, J = 7.2 Hz, 4H), 1.97–1.94 (m, 4H). LC/MS: m/z 418.1 (M+H)+(ES).

Example 19: Synthesis of compound 16, ethyl-7-(N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)sulfamoyl)-5 ,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine-3-carboxylate

[Step 1] Synthesis of Compound L1

7-(tert-butyl)-3-ethyl-5,6-dihydro-[1,2,4]triazolo(4,3-a)pyrazine-3,7(8H)-dicarboxylate (200 mg, 0.65 mmol) was dissolved in methanol (1 mL), and then 4 N HCl (1,4-dioxane solution, 1.5 mL) was added. Then, the reaction was stirred at room temperature for 4 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to remove the solvent and then dried under reduced pressure to obtain compound L1. It was used in the next reaction without additional purification.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 10.50 (br, 1H), 4.59 (s, 2H), 4.53 (t, J = 5.4 Hz, 2H), 4.40 (dd, J = 14.4, 6.8 Hz, 2H) ), 3.58 (s, 2H), 1.35 (t, J = 7.0 Hz, 3H).

[Step 2] Synthesis of Compound L2 (Compound 16)

After dissolving compound L1 (157mg, 0.65mmol) and pyridine (0.13mL, 1.60mmol) in dichloromethane (3mL), compound B1 (212mg, 0.67mmol) obtained in step 1 of Example 2 was dissolved at 0°C. added. Then, the reaction was stirred at room temperature for 4.5 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The reaction concentrate was purified by silica gel column chromatography (MC/MeOH), and solidified with dichloromethane and hexane to obtain the target compound L2 (Compound 16) (1.01mg, 0.3%) as a white solid.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 10.57 (br, 1H), 8.10 (br, 1H), 6.89 (s, 1H), 4.81 (s, 2H), 4.46-4.34 (m, 4H), 3.76 (s, 2H), 2.76 (t, J = 7.0 Hz, 4H), 2.46 (m, 4H), 1.88 (t, J = 7.4 Hz, 4H), 1.34 (t, J = 7.0 Hz, 3H). LC/MS: m/z 475.1 (M+H)+(ES).

Example 20: Synthesis of Compound 20, N-((2,4,5,6-tetrahydro-1H-cyclobuta[f]inden-3-yl)carbamoyl)-5,6-dihydro-[1 ,2,4]triazolo[4,3-a]pyrazine-7(8H)-sulfonamide

[Step 1] Synthesis of Compound M1

Chlorosulfonyl isocyanate (0.25mL, 2.8mmol) was added to isopropyl ether (4mL) and dissolved. Here, 2,4,5,6-tetrahydro-1H-cyclobuta)[1f]inden-3-amine (446mg, 2.8mmol) was dissolved in isopropyl ether (10mL) and added dropwise at -15°C. did The reaction was stirred at -15 °C for 1 hour. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain compound M1 (598mg, 71%) as a yellow oil.

[Step 2] Synthesis of Compound M2 (Compound 20)

Under a nitrogen atmosphere, compound M1 (50mg, 0.17mmol) and 5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine (21mg, 0.17mmol) were tetrahydro After adding and dissolving in furan (THF) (2 mL), sodium hydride (60% in mineral oil; 28.5 mg, 0.57 mmol) was added at -10 °C. The reaction was stirred at -10 °C for 1 hour 30 minutes. Thereafter, the reaction was terminated using an aqueous ammonium chloride solution, and the organic layer was extracted using ethyl acetate. The organic layer was dried using sodium sulfate and then concentrated under reduced pressure. The reaction concentrate was purified by silica gel column chromatography (MC/MeOH) to obtain the desired compound M2 (Compound 20) (1.2mg, 1.8%) as a yellow oil.

¹ H NMR (400 MHz, DMSO-d6) δ 10.55 (br, 1H), 8.51 (s, 1H), 8.12 (s, 1H), 6.71 (s, 1H), 4.71 (s, 2H), 4.13 (t , J = 5.6 Hz, 2H), 3.78 (t, J = 5.2 Hz, 2H), 2.93 (m, 2H), 2.88 (m, 2H), 2.81 (t, J = 7.6 Hz, 2H), 2.66 (t , J = 7.2 Hz, 2H), 1.99–1.93 (m, 2H). LC/MS: m/z 389.1 (M+H)+(ES).

<Experimental example>

Experimental Example 1. ELISA for interleukin-1β (IL-1β)

The mouse cell line J774A.1 (ATCC) was cultured in DMEM medium [DMEM (Hyclone) + 10% Fetal Bovine Serum (Hyclone) + 1% Penicillin-Streptomycin (Hyclone)].

Human THP-1 cells (ATCC) were cultured in RPMI medium [RPMI (Hyclone) + 10% Fetal Bovine Serum (Hyclone) + 1% Penicillin-Streptomycin (Hyclone) + 0.05 mM concentration of 2- and cultured in mercaptoethanol].

J774A.1 cells were dispensed in a 96-well plate (tissue culture plate, 96 well, Falcon) at a concentration of 100,000 cells/well, treated with LPS (Sigma) (2 μg/ml) for 4 h, and the example compounds (0.001 to 10,000 nM) were each treated for 30 min, and then treated with ATP (Sigma) (5 mM) for 1 h to induce inflammasome, and then the cell culture medium was used as a sample.

THP-1 cells were each dispensed at a concentration of 50,000 cells/well, differentiated by treatment with PMA (Sigma) (100 ng/ml) for 20 h, and then treated with LPS (Sigma) (1 μg/ml) for 4 h. Example compounds ( 0.001 ~ 10,000 nM) each was treated for 30 min, and then treated with ATP (Sigma) (5 mM) for 1 h to induce inflammasome, and then the cell culture medium was used as a sample.

IL-1β was measured using an ELISA kit (Invitrogen) according to the manufacturer's instructions, and absorbance was measured at 450 nm using a spectrophotometer microplate reader (BioTek, Synergy H4).

<Activity in J774A.1 or THP-1 (IC ₅₀ )>

A: less than 50 nM

B: 50 nM to 100 nM

C: greater than 100 nM to 1 uM

D: greater than 1uM to 10uM

E: greater than 10uM

[Table 4]

* NT: not tested

As can be seen in Table 4, it was confirmed that the compounds of the present invention or salts thereof can inhibit the release of IL-1β induced by NLRP3 inflammasome activity. That is, the compound of the present invention provides an inflammasome-related mechanism, and thus can be usefully used for the treatment of diseases caused by overactivation of the NLRP3 inflammasome.

Experimental Example 2. Pharmacokinetic test in mice

Each of Compound 3 and Compound 4 of the present application was dissolved in 10% DMSO/10% Cremophor EL/80% water for injection and then orally administered to 9-week-old BALB/c female mice at a dose of 10 mg/kg. Up to 8 hours after oral administration, blood was collected in a tube containing lithium heparin. Plasma was separated by centrifuging the tube at 6,000g, 4°C, and 5 minutes. Then, after pretreatment by protein precipitation method, the concentration of the test substance in plasma was quantitatively analyzed by LC-MS/MS system. From the drug concentration-time curve obtained from the mouse plasma test sample, the pharmacokinetic parameters AUC, Cmax, Tmax and t _1/2 were obtained by non-compartmental model method using the PK solution program, and the results are shown in Table 5 below.

[Table 5]

As above, the present invention has been described in detail, to those skilled in the art, it will be clear that these specific descriptions are only preferred embodiments, and the scope of the present invention is not limited thereby. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Claims (7)

A compound represented by Formula I, a stereoisomer thereof or a pharmaceutically acceptable salt thereof:
<Formula I>

In the above formula I,
X ₁ and X ₂ are each independently N or CR ₂ ;
R ₁ is H, C1-C5 alkyl, C1-C5 alkyl-OH, -(C1-C5 alkyl)-O-(C1-C5 alkyl), -(C1-C5 alkyl)-NH _2, -(C1-C5 alkyl)-NH-(C1-C5 alkyl), -(C1-C5 alkyl)-N(C1-C5 alkyl) ₂ , cycloalkyl, or heterocycloalkyl;
R ₂ is H, C1-C5 alkyl, C1-C5 alkyl-OH, halogen, CF ₃ , NRaRb or C(=O)O(C1-C5 alkyl);
Ra and Rb are each independently H, C1-C5 alkyl, -C(=O)(C1-C5 alkyl), C1-C5 alkyl-OH, or -S(=O) ₂ (C1-C5 alkyl);
m and n are each independently an integer from 0 to 2;
Z is CH ₂ , NR ₃ or O;
R ₃ is H or C1-C5 alkyl;
W ₁ is O or NRc, Rc is H or CN;
W ₂ is O or S;

Is

,

or

ego,
X ₃ is CR ₄ or N, where k is 0 or 1;
R ₄ is H or halogen;
R ₅ and R ₆ are each independently H or C1-C5 alkyl.
The method of claim 1, wherein the compound represented by Formula I is a compound represented by the following Formula Ia,
A compound represented by Formula I, a stereoisomer thereof or a pharmaceutically acceptable salt thereof:
<Formula Ia>

In Formula Ia, each of X ₁ , X ₂ , R ₁ , W ₁ , W ₂ , Z and A is the same as defined in Formula I.
The method of claim 1, wherein the compound represented by Formula I is a compound represented by Formula II below,
A compound represented by Formula I, a stereoisomer thereof or a pharmaceutically acceptable salt thereof:
<Formula II>

In Formula II above,
X ₁ and X ₂ are each independently N or CR ₂ ;
R ₁ is H, C1-C5 alkyl, C1-C5 alkyl-OH, -(C1-C5 alkyl)-O-(C1-C5 alkyl), -(C1-C5 alkyl)-NH _2, -(C1-C5 alkyl)-NH-(C1-C5 alkyl), -(C1-C5 alkyl)-N(C1-C5 alkyl) ₂ , cycloalkyl, or heterocycloalkyl;
R ₂ is H, C1-C5 alkyl, halogen, CF ₃ , NRaRb or C(=0)O(C1-C5 alkyl);
Ra and Rb are each independently H or C1-C5 alkyl;
Z is NR ₃ or O;
R ₃ is H or C1-C5 alkyl;

Is

,

or

ego,
X ₃ is CR ₄ , where k is 0 or 1;
R ₄ is H or halogen;
R ₅ and R ₆ are each independently H or C1-C5 alkyl.
Compounds listed in the table below, stereoisomers thereof or pharmaceutically acceptable salts thereof:

According to claim 4,
Compounds listed in the table below, stereoisomers thereof or pharmaceutically acceptable salts thereof:

Comprising the compound according to any one of claims 1 to 5, a stereoisomer thereof or a pharmaceutically acceptable salt thereof,
A pharmaceutical composition for the treatment or prevention of NLRP3 activity-related diseases.
According to claim 6,
The NLRP3 activity-related disease
Inflammation, autoimmune disease, cancer, infection, central nervous system disease, metabolic disease, cardiovascular disease, respiratory disease, liver disease, kidney disease, eye disease, skin disease, lymphatic conditions, psychological disorders, graft-versus-host disease, allodynia, wound and Including one or more diseases selected from scars,
pharmaceutical composition.