Aryl heterocyclic kv1.3 inhibitor, and preparation method therefor and use thereof

Abstract

The present invention provides a novel Kv1.3 channel (or Kv1.3) inhibitor, which can be used for preventing and / or treating Kv1.3 channel (or Kv1.3)-related diseases, including immune and inflammatory diseases, such as multiple sclerosis, inflammatory bowel disease, ulcerative colitis, Crohn's disease, rheumatoid arthritis, type I diabetes, psoriasis and asthma, spondylitis and periodontitis; and obesity, type 2 diabetes, renal fibrosis, Alzheimer's disease, and ischemic stroke.

Description

Aryl heterocyclic Kv1.3 inhibitors and their preparation method and use Technical Field

[0001] The present invention relates to a Kv1.3 inhibitor, a preparation method and use thereof. The inhibitor and a pharmaceutical composition containing the inhibitor can be used to prevent and / or treat Kv1.3-related diseases or conditions. Background Art

[0002] Kv1.3 channels (or Kv1.3) are voltage-gated potassium channels (Kv) that are highly expressed in macrophages, microglia, and effector memory T cells (TEMs). Kv1.3 channels can modulate cell membrane potential, indirectly influencing calcium signaling in TEMs. Effector memory T cells mediate a variety of immune and inflammatory diseases, such as multiple sclerosis, inflammatory bowel disease, rheumatoid arthritis, type 1 diabetes, psoriasis, and asthma.

[0003] Autoimmune diseases are a broad category of illnesses caused by the body's immune system attacking normal cells and tissues. Current treatment options typically include anti-inflammatory or immunosuppressant drugs, which can only alleviate symptoms and have serious toxic side effects. Studies have shown that effector memory T cells are closely related to the causes of autoimmune diseases (Proceedings of the National Academy of Sciences, 103, 46 (2006): 17414-17419). Selectively inhibiting the function of effector memory T cells can be used to treat autoimmune diseases while avoiding damage to protective immune responses and reducing treatment-induced side effects. Effector memory T cells express Kv1.3 in large quantities and rely on it to perform cellular functions. Therefore, selective Kv1.3 inhibitors are expected to become low-toxic and highly effective drugs for the treatment of autoimmune diseases.

[0004] Peptide toxins with Kv1.3 inhibitory properties have been isolated from scorpions and sea anemones. Dalazatide (ShK-186), derived from unnatural amino acids, has demonstrated promising therapeutic efficacy in a Phase Ib clinical trial for the treatment of psoriasis. However, poor subtype selectivity and the inability of peptides to be taken orally limit their application. Therefore, the development of highly selective, orally available small-molecule Kv1.3 inhibitors holds significant social and market value.

[0005] Summary of the Invention

[0006] The present invention provides novel Kv1.3 channel (or Kv1.3) inhibitors, which can be used to prevent and / or treat diseases associated with Kv1.3 channel (or Kv1.3).

[0007] According to one aspect of the present invention, the present invention provides a compound represented by formula (A) or a stereoisomer thereof, or a pharmaceutically acceptable salt or deuterated substance thereof:

[0008] in,

[0009] X1, X2, X3, X4, and X5 are independently selected from C and N, and two of them are N, so that A is a 5-membered heteroaryl group containing two N;

[0010] R1, R2, R3, R4 are independently selected from H, F, Cl, Br, CN, C 1-4 Straight-chain or branched alkyl, C 2-4 Straight-chain or branched alkenyl, C 2-4 Straight-chain or branched alkynyl, C 1-4 Straight-chain or branched alkyloxy, C 2-4 Straight-chain or branched alkenyloxy, C 2-4 Straight-chain or branched alkynyloxy, C 3-7 Cycloalkyl.

[0011] In a preferred embodiment, in formula (A):

[0012] R1, R2, R3, R4 are independently selected from H, F, Cl, Br, CN, C 1-4 Straight-chain or branched alkyl, C 2-4 Straight-chain or branched alkenyl, C 2-4 Straight-chain or branched-chain alkynyl.

[0013] In a preferred embodiment, in formula (A):

[0014] R1, R2, R3, and R4 are independently selected from H, F, Cl, and Br.

[0015] In a preferred embodiment, in formula (A):

[0016] R1 and R2 are independently selected from F, Cl, and Br, and R3 and R4 are H.

[0017] In a preferred embodiment, in formula (A):

[0018] X1 and X4 are N, and X2, X3, and X5 are C.

[0019] In a preferred embodiment, in formula (A):

[0020] X2 and X3 are N, and X1, X4, and X5 are C.

[0021] In a preferred embodiment, in formula (A):

[0022] X1 and X4 are N, X2, X3, and X5 are C;

[0023] R1 and R2 are independently selected from F, Cl, and Br, and R3 and R4 are H.

[0024] In a preferred embodiment, in formula (A):

[0025] X2 and X3 are N, X1, X4, and X5 are C;

[0026] R1 and R2 are independently selected from F, Cl, and Br, and R3 and R4 are H.

[0027] According to one aspect of the present invention, the present invention provides a compound represented by formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt or deuterated substance thereof:

[0028] in,

[0029] X1, X2, X3, X4, and X5 are independently selected from C and N, and two of them are N, so that A is a 5-membered heteroaryl group containing two N;

[0030] R1, R2, R3, R4 are independently selected from H, F, Cl, Br, CN, C 1-4 Straight-chain or branched alkyl, C 2-4 Straight-chain or branched alkenyl, C 2-4 Straight-chain or branched alkynyl, C 1-4 Straight-chain or branched alkyloxy, C 2-4 Straight-chain or branched alkenyloxy, C 2-4 Straight-chain or branched alkynyloxy, C 3-7 Cycloalkyl.

[0031] In a preferred embodiment, in formula (I):

[0032] R1, R2, R3, R4 are independently selected from H, F, Cl, Br, CN, C 1-4 Straight-chain or branched alkyl, C 2-4 Straight-chain or branched alkenyl, C 2-4 Straight-chain or branched-chain alkynyl.

[0033] In a preferred embodiment, in formula (I):

[0034] R1, R2, R3, and R4 are independently selected from H, F, Cl, and Br.

[0035] In a preferred embodiment, in formula (I):

[0036] X1 and X4 are N, X2, X3, and X5 are C;

[0037] R1 and R2 are independently selected from F, Cl, and Br, and R3 and R4 are H.

[0038] In a preferred embodiment, in formula (I):

[0039] X2 and X3 are N, X1, X4, and X5 are C;

[0040] R1 and R2 are independently selected from F, Cl, and Br, and R3 and R4 are H.

[0041] According to one aspect of the present invention, the present invention provides a compound represented by formula (II) or a stereoisomer thereof, or a pharmaceutically acceptable salt or deuterated substance thereof:

[0042] in,

[0043] X1, X2, X3, X4, and X5 are independently selected from C and N, and two of them are N, so that A is a 5-membered heteroaryl group containing two N;

[0044] R1, R2, R3, R4 are independently selected from H, F, Cl, Br, CN, C 1-4 Straight-chain or branched alkyl, C 2-4 Straight-chain or branched alkenyl, C 2-4 Straight-chain or branched alkynyl, C 1-4 Straight-chain or branched alkyloxy, C 2-4 Straight-chain or branched alkenyloxy, C 2-4 Straight-chain or branched alkynyloxy, C 3-7 Cycloalkyl.

[0045] In a preferred embodiment, in formula (II):

[0046] R1, R2, R3, R4 are independently selected from H, F, Cl, Br, CN, C 1-4 Straight-chain or branched alkyl, C 2-4 Straight-chain or branched alkenyl, C 2-4 Straight-chain or branched-chain alkynyl.

[0047] In a preferred embodiment, in formula (II):

[0048] R1, R2, R3, and R4 are independently selected from H, F, Cl, and Br.

[0049] In a preferred embodiment, in formula (II):

[0050] X2 and X3 are N, X1, X4, and X5 are C;

[0051] R1 and R2 are independently selected from F, Cl, and Br, and R3 and R4 are H.

[0052] In a preferred embodiment, the compound of the present invention is selected from the following compounds or pharmaceutically acceptable salts or deuterated derivatives thereof:

[0053] In the present invention, although substituents are disclosed in groups or ranges, the groups or ranges of the present invention specifically refer to each specific group they cover. For example, the term "C 1-4 The term "alkyl" specifically refers to the independently disclosed methyl (ie, C1 alkyl), ethyl (ie, C2 alkyl), propyl (ie, C3 alkyl), and butyl (ie, C4 alkyl).

[0054] In the present invention, "C 3-7 Examples of "cycloalkyl" include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl.

[0055] In the present invention, examples of the "5-membered heteroaryl group containing 2 N atoms" include pyrazole and imidazole.

[0056] The compounds of the present invention may be asymmetric, for example, having one or more stereocenters. All stereoisomers, such as enantiomers and diastereomers, are included within the scope of the present invention unless otherwise indicated. In the present invention, compounds containing asymmetrically substituted carbon atoms can be isolated in either optically active or racemic forms. Various methods for preparing optically active forms are known in the art, for example, by resolving a racemic mixture or by stereoselective synthesis.

[0057] The present invention also includes pharmaceutically acceptable salts of the compounds. Pharmaceutically acceptable salts of the compounds of the present invention can be prepared by reacting with non-toxic inorganic or organic acids. Inorganic acids include hydrochloric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, sulfuric acid, nitric acid, hydrogen sulfate, boric acid, and hemisulfuric acid; organic acids include formic acid, acetic acid, propionic acid, butyric acid, valeric acid, hexanoic acid, heptanoic acid, undecanoic acid, palmitic acid, stearic acid, oleic acid, oxalic acid, malonic acid, adipic acid, lactic acid, malic acid, maleic acid, hippuric acid, tartaric acid, citric acid, succinic acid, ascorbic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, benzoic acid, camphoric acid, camphorsulfonic acid, citric acid, fumaric acid, gluconic acid, galacturonic acid, and dodecylsulfuric acid; and various amino acids.

[0058] The present invention also includes hydrates and solvates of the compounds.

[0059] The present invention also includes all forms of the compounds wherein the atoms are isotopes. Isotopes include all atoms having the same atomic number but different mass numbers. For example, isotopes of hydrogen include deuterium.

[0060] The present invention also includes prodrugs of the compounds. "Prodrugs" refer to compounds obtained by structural modification of the compounds that are inactive or less active outside the patient's body, but release the compound through enzymatic or non-enzymatic conversion in the patient's body to exert its pharmacological effect.

[0061] It will be appreciated by those skilled in the art that the compounds of the present invention can be prepared by various methods disclosed in the literature. The compounds of the present invention can be prepared by reacting in a suitable solvent. A person skilled in the art of organic synthesis can easily select a suitable solvent that does not substantially react with the reactants, intermediates, or products. The reaction can be carried out in a solvent or in a mixture of more than one solvent. The compounds of the present invention can be prepared by reacting at a suitable temperature, for example, between the freezing temperature of the solvent and the boiling temperature of the solvent. The method for preparing the compounds of the present invention involves protecting and deprotecting various chemical groups. A person skilled in the art of organic synthesis can easily determine whether chemical groups need to be protected and deprotected and select suitable protecting groups. The reaction for preparing the compounds of the present invention can be monitored using any method known in the art, such as nuclear magnetic resonance spectroscopy, infrared spectroscopy, mass spectrometry, chromatography, etc. The following schemes 1-2 describe synthetic routes that can be used to synthesize the compounds of the present invention, for example, compounds having the structure of Formula I.

[0062] As shown in Scheme 1, compounds 1-a, 1-b, and 1-c can be prepared by any method known in the art and / or purchased commercially. As shown in Scheme 1, PG refers to a protecting group. Non-limiting examples of protecting groups include methyl, allyl, acetyl, tert-butyloxycarbonyl, benzyl, or other protecting groups known in the art to be suitable as OH protecting groups. Other substituents are defined herein. As shown in Scheme 1, 1-a can be subjected to a two-step reaction of phenolic hydroxyl protection and formylation to produce 1-c; 1-c can be subjected to a Wittig reaction, nitromethane substitution, nitro group reduction, and intramolecular cyclization to produce 1-g; and 1-h can be reacted with methyloxonium tetrafluoroborate to produce the intermediate 1-h. 1-h can be reacted with aminoacetaldehyde dimethyl acetal through substitution and ring closure to produce 1-j, which can then be subjected to bromination to produce 1-k. 1-k can be subjected to a Suzuki reaction or Stille reaction to produce 1-l, or it can be substituted with a hydroxyl- or amino-containing compound to produce 1-l, and deprotection to produce the final product 1-m.

[0063] Compound 2-c, as shown in Scheme 2, can be prepared by any method known in the art and / or commercially available. As shown in Scheme 2, 2-a reacts with a triflate to produce 2-b, which is then reacted with 2-c via a Suzuki reaction to produce 2-d, which is then reacted with DMFDMA to produce 2-e. This is then subjected to a two-step ring closure with hydrazine hydrate and reduction to produce 2-g. 2-g undergoes a Mitsunobu reaction or substitution reaction to produce 2-h, which is then deprotected to produce the final product 2-i. Intermediate 2-f can also be ring-closed with a substituted hydrazine to produce 2-j, which is then reduced and deprotected to produce the final product 2-l.

[0064] The compounds of the present invention are capable of inhibiting Kv1.3. Therefore, according to another aspect of the present invention, there is provided a method for inhibiting Kv1.3 using the compounds of the present invention.

[0065] According to another aspect of the present invention, a method for preventing and / or treating Kv1.3-related diseases is provided, wherein a therapeutically and / or preventively effective amount of a compound of the present invention or a pharmaceutical composition containing the compound of the present invention is administered to an individual in need thereof.

[0066] Kv1.3-related diseases include any disease that is directly and / or indirectly related to the expression and / or activity of Kv1.3, for example, a disease that can be prevented and / or treated by inhibiting Kv1.3.

[0067] Kv1.3-related diseases include immune and inflammatory diseases such as multiple sclerosis, inflammatory bowel disease, ulcerative colitis, Crohn's disease, rheumatoid arthritis, type 1 diabetes, psoriasis and asthma, spondylitis, periodontitis; as well as obesity, type 2 diabetes, renal fibrosis, Alzheimer's disease, ischemic stroke, etc.

[0068] According to another aspect of the present invention, the present invention provides use of the compound of the present invention in preparing a Kv1.3 inhibitor.

[0069] According to another aspect of the present invention, the present invention provides use of the compound of the present invention in the preparation of a medicament for preventing and / or treating Kv1.3-related diseases.

[0070] When the compound of the present invention is used to prevent and / or treat Kv1.3-related diseases, the compound of the present invention can be administered in the form of a pharmaceutical composition. Therefore, according to another aspect of the present invention, the present invention provides a pharmaceutical composition comprising the compound of the present invention and a pharmaceutically acceptable carrier.

[0071] It will be appreciated by those skilled in the art that the pharmaceutical compositions of the present invention can be prepared by various methods disclosed in the literature. The compounds or pharmaceutical compositions of the present invention can be administered by a variety of routes, depending on the area where local or systemic treatment is needed and the area where treatment is needed. For example, the compositions can be administered orally, parenterally (e.g., intravenously, intraarterially, subcutaneously, intraperitoneally, intramuscularly or by infusion), intracranially, e.g., intrathecally or intraventricularly, transdermally, ophthalmically, nasally, vaginally, rectally, or pulmonary (e.g., by inhalation or insufflation of a powder or aerosol).

[0072] For oral administration, the pharmaceutical compositions of the present invention are typically provided in the form of tablets, capsules, or solutions. Tablets may comprise a compound of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. The carrier includes, but is not limited to, a diluent, a disintegrant, a binder, a lubricant, a colorant, or a preservative. Capsules include hard capsules and soft capsules. For parenteral administration, the pharmaceutical compositions of the present invention may be administered by intravenous, intramuscular, or subcutaneous injection. They are typically provided as sterile aqueous solutions or suspensions or lyophilized powders, and are adjusted for appropriate pH and isotonicity.

[0073] The effective amount of the compounds of the present invention can be determined based on the specific use of the treatment, the mode of administration, and the condition of the individual in need, such as the patient. Those skilled in the art are capable of determining the effective amount of the compounds of the present invention. Typical dosage ranges are, for example, 1 μg / kg / day to 1000 mg / kg / day.

[0074] When preventing and / or treating Kv1.3-related diseases, the compounds of the present invention may be used in combination with one or more other drugs. Other drugs include but are not limited to prednisone, methylprednisolone, ocrelizumab, natalizumab, alemtuzumab, beta interferon, glatiramer, fingolimod, dimethyl fumarate, desiroximet, teriflunomide, siponimod, cladribine, mesalamine, balaxazine, olsalazine, azathioprine, mercaptopurine, methotrexate, infliximab, adalimumab, golimumab, becelizumab, vedolizumab, Ustekinumab, methotrexate, leflunomide, hydroxychloroquine, sulfasalazine, abatacept, adalimumab, anakinra, becelizumab, etanercept, golimumab, infliximab, rituximab, salilukumab, tocilizumab, baricitinib, tofacitinib, upadacitinib, insulin glulisine, insulin lispro, insulin aspart, insulin glargine, insulin detemir, insulin degludec, fluticasone, budesonide, mometasone furoate, beclomethasone, ciclesonide, montelukast, zafirlukast, zileuton, salmeterol, omalizumab, mepolizumab, benralizumab, reslizumab, etc. BRIEF DESCRIPTION OF THE DRAWINGS

[0075] Figure 1 Ellipsoid diagram of the molecular structure of compound (S)-4-(6-(allyloxy)-2,3-dichlorophenyl)pyrrolidin-2-one

[0076] Figure 2 Ellipsoid diagram of the molecular structure of compound (R)-4-(6-(allyloxy)-2,3-dichlorophenyl)pyrrolidin-2-one

[0077] Figure 3 NOESY spectrum of Example 3

[0078] FIG4 NOESY spectrum of Example 4

[0079] Figure 5 NOESY spectrum of Example 5 DETAILED DESCRIPTION

[0080] Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiments are only used to illustrate the present invention and are not used in limiting the scope of the present invention.In addition, should be understood that after reading the content of the present invention being recorded, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms fall within the limited scope of the present invention equally.

[0081] Intermediate 1. 3-(6-(allyloxy)-2,3-dichlorophenyl)-5-methoxy-3,4-dihydro-2H-pyrrole

[0082] Synthesis of 4-(allyloxy)-1,2-dichlorobenzene (2):

[0083] 3,4-Dichlorophenol (120.0 g, 0.74 mol) and potassium carbonate (253.9 g, 1.84 mol) were mixed in N,N-dimethylformamide (1.5 L). Allyl bromide (106.9 g, 0.88 mol) was added portionwise at 0°C. The resulting mixture was stirred at room temperature for 16 hours. After completion of the reaction, it was diluted with water (1500 mL) and extracted twice with ethyl acetate (1500 mL). The combined organic phases were washed with saturated brine (1500 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 20:1) to yield a yellow solid (120.0 g, 68% yield).

[0084] Synthesis of 6-(allyloxy)-2,3-dichlorobenzaldehyde (3):

[0085] 4-(Allyloxy)-1,2-dichlorobenzene (120.0 g, 0.59 mol) was dissolved in tetrahydrofuran (1200 mL) at -78°C, and n-butyllithium (260.0 mL, 0.65 mol, 2.5 M solution in n-hexane) was added dropwise over 30 minutes. N,N-dimethylformamide (86.4 g, 1.18 mol) was then added at the same temperature, and the resulting mixture was stirred at the same temperature for another 3 hours. After completion, the reaction was quenched with saturated aqueous ammonium chloride (1000 mL) and extracted twice with ethyl acetate (3000 mL). The combined organic phases were washed with saturated brine (1500 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 15:1) to afford a pale yellow solid (139.5 g, 86% yield).

[0086] 1 H NMR (400MHz, DMSO-d6) δ10.36(s,1H),7.83(d,J=9.1Hz,1H),7.25(d,J=9.1Hz,1H),6.13–5.96(m ,1H),5.46(m,J=17.3,1.7Hz,1H),5.31(m,J=10.6,3.0,1.4Hz,1H),4.74(dt,J=4.9,1.5Hz,2H).

[0087] Synthesis of ethyl (E)-3-(6-(allyloxy)-2,3-dichlorophenyl)acrylate (4):

[0088] To a solution of ethyl 2-(dimethoxyphosphoryl)acetate (142.6 g, 0.73 mol) in tetrahydrofuran (2000 mL) at 0°C was added sodium hydride (29.1 g, 0.73 mol, 60% in mineral oil). After stirring for 30 minutes, 6-(allyloxy)-2,3-dichlorobenzaldehyde (139.5 g, 0.61 mol) was added and stirred at 0°C for 4 hours. After completion of the reaction, the mixture was diluted with water (2000 mL) and extracted with ethyl acetate (4000 mL). The organic phase was washed with saturated brine (2000 mL) and dried over anhydrous sodium sulfate. The residue was concentrated under reduced pressure and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 5:1) to afford a yellow solid (170.0 g, 79% yield).

[0089] 1H NMR (400MHz, DMSO-d6) δ7.84(d,J=16.2Hz,1H),7.64(d,J=9.1Hz,1H),7.17(d,J=9.2Hz,1H),6.81(d,J=16.2Hz,1H ),6.13–6.00(m,1H),5.47–5.25(m,2H),4.74(dt,J=5.2,1.5Hz,2H),4.21(q,J=7.1Hz,2H),1.26(t,J=7.1Hz,3H).

[0090] Synthesis of ethyl 3-(6-(allyloxy)-2,3-dichlorophenyl)-4-nitrobutyrate (5):

[0091] To a reaction flask containing nitromethane (2000 mL) at room temperature were added ethyl (E)-3-(6-(allyloxy)-2,3-dichlorophenyl)acrylate (170.0 g, 0.56 mol) and 1,8-diazabicyclo[5.4.0]undec-7-ene (103.1 g, 0.68 mol). The reaction mixture was stirred at 60°C for 16 hours. After completion, the mixture was diluted with water (1000 mL) and extracted with ethyl acetate (3000 mL). The organic phase was washed with saturated brine (1000 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 5:1) to afford a light yellow oil (174.0 g, 72% yield).

[0092] 1 H NMR (400MHz, DMSO-d6) δ7.53(d,J=9.0Hz,1H),7.06(d,J=9.1Hz,1H),6.07(ddd,J=15.9,10.6,5.2Hz,1H),5.41(d,J=17.2Hz,1H),5.33(dd,J=10. 6,1.1Hz,1H),5.00(ddd,J=19.0,13.0,7.5Hz,2H),4.69(d,J=4.4Hz,3H) ,4.04–3.91(m,2H),2.89(dq,J=17.6,8.7Hz,2H),1.06(t,J=7.1Hz,3H).

[0093] Synthesis of ethyl 3-(6-(allyloxy)-2,3-dichlorophenyl)-4-aminobutyrate (6):

[0094] Ethyl 3-(6-(allyloxy)-2,3-dichlorophenyl)-4-nitrobutyrate (174.0 g, 0.48 mol) and zinc powder (314.2 g, 4.80 mol) were added to a reaction flask containing glacial acetic acid (2000 mL) at room temperature. The mixture was stirred at 25°C for 3 hours. After completion of the reaction, the mixture was filtered, and the filter cake was washed twice with ethyl acetate. The filtrate was collected and concentrated under reduced pressure to obtain 200.0 g of crude product, which was used directly in the next step without purification.

[0095] LCMS [M+H] + :332.1.

[0096] Synthesis of 4-(6-(allyloxy)-2,3-dichlorophenyl)pyrrolidin-2-one (7):

[0097] Crude ethyl 3-(6-(allyloxy)-2,3-dichlorophenyl)-4-aminobutyrate (200.0 g, 0.45 mol) and potassium carbonate (311.5 g, 2.26 mol) were added to methanol (2000 mL) and stirred at 50°C for 3 hours. After the reaction, the mixture was diluted with water (1000 mL) and extracted three times with ethyl acetate (3000 mL). The organic phases were combined and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. Purification by silica gel column chromatography (petroleum ether:ethyl acetate = 5:1) afforded a white solid (105.0 g, 69% yield).

[0098] LCMS [M+H] + :286.0.

[0099] 1 H NMR (400MHz, DMSO-d6) δ7.75(s,1H),7.51(d,J=9.0Hz,1H),7.08(d,J=9.0Hz,1 H),6.01(ddt,J=17.2,10.4,5.0Hz,1H),5.44–5.35(m,1H),5.26(dd,J=10.6,1. 5Hz,1H),4.64(dd,J=3.6,1.4Hz,2H),4.33(dd,J=18.4,9.2Hz,1H),3.43(dt,J =16.7,8.9Hz,2H),2.54(dd,J=16.7,9.0Hz,1H),2.36(dd,J=16.5,10.7Hz,1H).

[0100] Synthesis of 3-(6-(allyloxy)-2,3-dichlorophenyl)-5-methoxy-3,4-dihydro-2H-pyrrole (8):

[0101] Dissolve 4-(6-(allyloxy)-2,3-dichlorophenyl)pyrrolidin-2-one (500.0 mg, 1.75 mmol) in dichloromethane (5 mL), add trimethyloxonium tetrafluoroborate (311.6 mg, 2.11 mmol), and stir the mixture at room temperature under nitrogen for 3 hours. After the reaction, spin dry the mixture and use the crude product in the next step without purification.

[0102] Intermediate 2. (S)-3-(6-(allyloxy)-2,3-dichlorophenyl)-5-methoxy-3,4-dihydro-2H-pyrrole

[0103] Preparation of (S)-4-(6-(allyloxy)-2,3-dichlorophenyl)pyrrolidin-2-one (2):

[0104] 4-(6-(Allyloxy)-2,3-dichlorophenyl)pyrrolidin-2-one (10 g) was purified by chiral preparation (gradient column: CHIRALPAK IC 250 mm×4.6 mm, 5 μm, flow rate: 3 mL / min, gradient: 40% IPA (NH4OH 0.2%): 60% CO2).

[0105] The first compound obtained by SFC separation was compound 2 (4.5 g, yield 45%), which was analyzed by SFC chromatographic conditions, RT = 4.51 min. LC-MS: [M+H] + :286.0.

[0106] The second compound obtained by SFC separation was compound 3 (4.3 g, yield 43%), which was analyzed by SFC chromatographic conditions, RT = 6.5 min. LC-MS: [M+H] + :286.0.

[0107] Single crystal X-ray diffraction analysis of compounds 2 and 3

[0108] Preparation of single crystals: Single crystals of Compound 2 and Compound 3 were obtained by solvent diffusion crystallization under ethyl acetate / n-hexane conditions and cultured at room temperature for 2 days.

[0109] Single crystal testing instrument model: Bruker D8 Venture

[0110] Instrument parameters:

[0111] Light source: Cu target X-ray: Cu-Kα

[0112] Detector: CMOS area detector Resolution:

[0113] Current and voltage: 50kV, 1.2mA Exposure time: 5s

[0114] Distance from surface detector to sample: 40mm Test temperature: 100(2)K

[0115] Structural analysis and refinement process:

[0116] After the diffraction data were integrated and reduced using the SAINT program, the data were empirically corrected for absorption using the SADABS program. The single crystal structure was directly resolved using SHELXT2014, and the structure was refined using the least squares method. The hydrogen atom refinement process was obtained using isotropic calculations. The hydrogen atoms on CH were obtained by computational hydrogenation and refined using the riding model. The Flack constant of the single crystal of compound 2 was 0.087 (13), and C9 was in the S configuration; the Flack constant of the single crystal of compound 3 was 0.073 (13), and C3 was in the R configuration. The molecular stereoscopic ellipsoid diagrams of compounds 2 and 3 are shown in Figures 1 and 2. The crystal structure data of compound 2 are shown in Table 1, and the crystal structure data of compound 3 are shown in Table 2.

[0117] Table 1. Crystal data of compound 2

[0118] Table 2. Crystal data of compound 3

[0119] Synthesis of (S)-3-(6-(allyloxy)-2,3-dichlorophenyl)-5-methoxy-3,4-dihydro-2H-pyrrole (4):

[0120] Dissolve (S)-4-(6-(allyloxy)-2,3-dichlorophenyl)pyrrolidin-2-one (1.0 g, 3.5 mmol) in dichloromethane (10 mL). Add trimethyloxonium tetrafluoroborate (623.4 mg, 4.23 mmol). Stir the mixture at room temperature under nitrogen for 3 hours. After the reaction, spin dry the mixture and use the crude product in the next step without purification.

[0121] Example 1-2. 3,4-Dichloro-2-((S)-3-((R)-pyrrolidin-3-yl)-6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-6-yl)phenol and 3,4-Dichloro-2-((S)-3-((S)-pyrrolidin-3-yl)-6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-6-yl)phenol

[0122] Synthesis of (S)-3-(6-(allyloxy)-2,3-dichlorophenyl)-N-(2,2-dimethoxyethyl)-3,4-dihydro-2H-pyrrol-5-amine (2):

[0123] (S)-3-(6-(allyloxy)-2,3-dichlorophenyl)-5-methoxy-3,4-dihydro-2H-pyrrole (10.0 g, 33.33 mmol) and 2,2-dimethoxyethan-1-amine (4.2 g, 39.95 mmol) were added to methanol (100.0 mL) at room temperature, and the resulting solution was stirred at 70°C for 4 hours. After the reaction, the reaction solution was concentrated under reduced pressure, and the crude product was used directly in the next step without purification.

[0124] LC-MS: [M+H] + :373.0.

[0125] Synthesis of (S)-6-(6-(allyloxy)-2,3-dichlorophenyl)-6,7-dihydro-5H-pyrrolo[1,2-a]imidazole (3):

[0126] The crude product from the previous step was dissolved in formic acid (100 mL) at room temperature, and the resulting mixture was stirred at 130°C for 4 hours. After the reaction, the formic acid was removed by concentration under reduced pressure. The resulting residue was diluted with saturated aqueous sodium bicarbonate (200.0 mL) and extracted three times with ethyl acetate (500.0 mL). The combined organic phases were washed with saturated sodium chloride (200.0 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated by silica gel column chromatography (dichloromethane:methanol = 20:1) to obtain the desired product (8.1 g, 66%).

[0127] LC-MS: [M+H] + :308.9.

[0128] Synthesis of (S)-6-(6-(allyloxy)-2,3-dichlorophenyl)-3-bromo-6,7-dihydro-5H-pyrrolo[1,2-a]imidazole (4):

[0129] (S)-6-(6-(allyloxy)-2,3-dichlorophenyl)-6,7-dihydro-5H-pyrrolo[1,2-a]imidazole (6.5 g, 0.021 mol) was dissolved in acetonitrile (65 mL) at -10°C, followed by the addition of N-bromosuccinimide (3.4 g, 0.019 mol) in portions. The reaction was stirred at this temperature for 30 minutes. After completion, the reaction was quenched with sodium thiosulfate solution (100 mL) and extracted twice with ethyl acetate (200 mL). The combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane:methanol = 15:1) to afford a pale yellow solid (2.4 g, 25% yield).

[0130] LC-MS: [M+H] + :386.7.

[0131] Synthesis of (S)-3-(6-(2,3-dichloro-6-hydroxyphenyl)-6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-3-yl)-2,5-dihydro-1H-pyrrole-1-carboxylic acid tert-butyl ester (5):

[0132] To a reaction flask containing dioxane and water (10 mL, 1 mL) were added (S)-6-(6-(allyloxy)-2,3-dichlorophenyl)-3-bromo-6,7-dihydro-5H-pyrrolo[1,2-a]imidazole (370.0 mg, 0.95 mmol), tert-butyl 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-2,5-dihydro-1H-pyrrole-1-carboxylate (562.8 mg, 1.91 mmol), sodium carbonate (202.1 mg, 1.91 mmol) and tetrakistriphenylphosphine palladium (110.2 mg, 0.09 mol) at room temperature, and the mixture was stirred at 110°C for 16 hours. After completion of the reaction, the mixture was diluted with water (30 mL) and extracted twice with ethyl acetate (50 mL). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using dichloromethane:methanol = 10:1 as the eluent to afford tert-butyl (S)-3-(6-(2,3-dichloro-6-hydroxyphenyl)-6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-3-yl)-2,5-dihydro-1H-pyrrole-1-carboxylate as an off-white solid (190 mg, 38% yield).

[0133] LC-MS: [M+H] + :435.7.

[0134] Synthesis of tert-butyl 3-((S)-6-(2,3-dichloro-6-hydroxyphenyl)-6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-3-yl)pyrrolidine-1-carboxylate (6):

[0135] (S)-tert-Butyl 3-(6-(2,3-dichloro-6-hydroxyphenyl)-6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-3-yl)-2,5-dihydro-1H-pyrrole-1-carboxylate (180.0 mg, 0.41 mol) and platinum dioxide (90.0 mg) were added to methanol and dichloromethane (5 / 5 mL) and stirred at 25°C under a hydrogen atmosphere for 16 hours. After the reaction, the mixture was filtered and the filtrate was concentrated under reduced pressure to give the desired product (120.0 mg, 66%), which was used directly in the next step without purification.

[0136] LC-MS: [M+H] + :438.0.

[0137] Synthesis of 3,4-dichloro-2-((6S)-3-(pyrrolidin-3-yl)-6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-6-yl)phenol (7):

[0138] Tert-butyl 3-((S)-6-(2,3-dichloro-6-hydroxyphenyl)-6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-3-yl)pyrrolidine-1-carboxylate (120 mg) was added to dichloromethane (5.0 mL) at room temperature. Dioxane hydrochloride (1.0 mL) was slowly added to the reaction solution and stirred for 3 hours. After completion of the reaction, the residue was concentrated under reduced pressure and purified by preparative chromatography (chromatographic column: -Gemini-C18 150 x 21.2 mm, 5 um mobile phase: ACN-H2O (0.1% FA) gradient: 2-30) to give the title product (50.0 mg, 54%).

[0139] LC-MS: [M+H] + :338.0.

[0140] Preparation of 3,4-dichloro-2-((S)-3-((R)-pyrrolidin-3-yl)-6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-6-yl)phenol and 3,4-dichloro-2-((S)-3-((S)-pyrrolidin-3-yl)-6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-6-yl)phenol (8 / 9):

[0141] 3,4-Dichloro-2-((6S)-3-(pyrrolidin-3-yl)-6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-6-yl)phenol (50.0 mg, 0.14 mmol) was chirally prepared (chromatographic column: CHIRALPAK AD-H 250 mm 20 mm, 5 μm, mobile phase: 40% EtOH (NH4OH 0.2%)) to give a pair of enantiomers.

[0142] The first compound obtained by SFC resolution was numbered as Example 1 (15.5 mg, yield 31.0%), which was analyzed by SFC analysis chromatographic conditions, RT = 2.48 min.

[0143] LC-MS: [M+H] + :338.0.

[0144] 1 H NMR (400MHz, DMSO-d6) δ8.36 (s, 1H), 7.31 (d, J = 8.8Hz, 1H), 7.06 (s, 1H), 6.81 (d ,J=8.8Hz,1H),5.16–5.04(m,1H),4.38(t,J=9.8Hz,1H),4.28(dd,J=10.0,7.5H z,1H),3.75–3.59(m,2H),3.51(ddd,J=12.6,8.2,4.7Hz,1H),3.39(dt,J=11.7, 8.4Hz,1H),3.30–3.18(m,3H),2.49(td,J=12.3,7.0Hz,1H),2.20–2.08(m,1H).

[0145] The second compound obtained by SFC separation was numbered as Example 2 (14.1 mg, yield 28.2%), which was analyzed by SFC analysis chromatographic conditions, RT = 4.37 min.

[0146] LC-MS: [M+H] + :338.0.

[0147] 1H NMR (400MHz, DMSO-d6) δ8.37(s,1H),7.31(d,J=8.8Hz,1H),7.10(s,1H),6.81(d,J=8.8Hz,1H),5.09(dd,J=16.5,8.1Hz,1H),4.35(dt,J=17.1,9.8Hz ,2H),3.81–3.59(m,2H),3.52(s,1H),3.39(dd,J=17.4,9.2Hz,1H),3.29(d d,J=10.7,5.1Hz,3H),2.49(d,J=6.0Hz,1H),2.09(dd,J=12.4,8.1Hz,1H).

[0148] Example 3-5. (R)-3-((R)-5-(2,3-dichloro-6-methoxyphenyl)-5,6-dihydrocyclopenta[c]pyrazol-1(4H)-yl)pyrrolidine-1-carboxylic acid tert-butyl ester, (R)-3-((S)-5-(2,3-dichloro-6-methoxyphenyl)-5,6-dihydrocyclopenta[c]pyrazol-1(4H)-yl)pyrrolidine-1-carboxylic acid tert-butyl ester, and (3R)-3-(5-(2,3-dichloro-6-methoxyphenyl)-5,6-dihydrocyclopenta[c]pyrazol-2(4H)-yl)pyrrolidine-1-carboxylic acid tert-butyl ester

[0149] Synthesis of 3-oxocyclopent-1-en-1-yl trifluoromethanesulfonate (2):

[0150] To a solution of 1,3-cyclopentanedione (10.0 g, 0.10 mol) in tetrahydrofuran (150.0 mL) was added triethylamine (20.5 g, 0.20 mol) at 0°C. After stirring for 10 minutes, trifluoromethanesulfonic anhydride (34.5 g, 0.12 mol) was slowly added dropwise to the mixture while maintaining the temperature at 0°C. After the addition was complete, the mixture was warmed to room temperature and stirred for 3 hours. After TLC was completed, the solvent of the mixture was concentrated under reduced pressure at 30°C. The residue was wet-eluted with a sample / eluent (petroleum ether:ethyl acetate = 20:1). The resulting product was concentrated under reduced pressure at 30°C, and most of the solvent was used directly in the next step.

[0151] Synthesis of 3-(2,3-dichloro-6-methoxyphenyl)cyclopent-2-en-1-one (3):

[0152] To a mixed solution of (2,3-dichloro-6-methoxyphenyl)boronic acid (10.0 g, 0.05 mol) (dioxane:water = 5:1, 300 mL) was added 3-oxocyclopent-1-en-1-yl trifluoromethanesulfonate (20.0 g, 0.09 mol), potassium carbonate (16.2 g, 0.12 mol), and tetrakistriphenylphosphine palladium (5.2 g, 0.004 mol). The mixture was stirred at 80°C for 16 hours under nitrogen. After completion of the reaction, the mixture was diluted with water (100.0 mL) and extracted twice with ethyl acetate (100.0 mL). The combined organic phases were washed with saturated brine (200.0 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 3:1) to obtain the desired product (4.0 g, 34% yield).

[0153] LC-MS: [M+H] + :257.0.

[0154] Synthesis of (E)-3-(2,3-dichloro-6-methoxyphenyl)-5-((dimethylamino)methylene)cyclopent-2-en-1-one (4):

[0155] 3-(2,3-Dichloro-6-methoxyphenyl)cyclopent-2-en-1-one (300.0 mg, 1.17 mmol) was added to a solution of N,N-dimethylformamide dimethyl acetal (4.0 mL) at room temperature, and the mixture was stirred at 110°C for 3 hours. After the reaction, the mixture was directly spin-dried to obtain a yellow oil (300.0 mg of a mixture of crude product and starting material, yield: 50%), which was used directly in the next step.

[0156] LC-MS: [M+H] + :312.0.

[0157] Synthesis of 5-(2,3-dichloro-6-methoxyphenyl)-1,4-dihydrocyclopentadienyl[c]pyrazole (5):

[0158] To a solution of (E)-3-(2,3-dichloro-6-methoxyphenyl)-5-((dimethylamino)methylene)cyclopent-2-en-1-one (300.0 mg, 0.96 mmol) and hydrazine hydrochloride (302.84 mg, 4.42 mmol) in ethanol (4.0 mL) was added triethylamine (0.25 mL). The mixture was stirred at 80°C for 1 hour. After completion of the reaction, the filtrate was concentrated under reduced pressure to dryness. The resulting crude product was purified by silica gel chromatography (dichloromethane:methanol = 30:1) to afford a white solid (100.0 mg, 37% yield).

[0159] LC-MS: [M+H] + :281.0.

[0160] Synthesis of 5-(2,3-dichloro-6-methoxyphenyl)-1,4,5,6-tetrahydrocyclopenta[c]pyrazole (6):

[0161] Platinum dioxide (701.1 mg, 3.09 mmol) was added to a solution of 5-(2,3-dichloro-6-methoxyphenyl)-1,4-dihydrocyclopenta[c]pyrazole (700.0 mg, 2.49 mmol) in dichloromethane (35.0 mL). The mixture was stirred at room temperature under a hydrogen atmosphere for 7 days. After the reaction, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel chromatography (dichloromethane:methanol = 40:1) to afford a yellow solid (280.0 mg, 40% yield).

[0162] LC-MS: [M+H] + :283.0.

[0163] Synthesis of tert-butyl (R)-3-((R)-5-(2,3-dichloro-6-methoxyphenyl)-5,6-dihydrocyclopenta[c]pyrazol-1(4H)-yl)pyrrolidine-1-carboxylate, tert-butyl (R)-3-((S)-5-(2,3-dichloro-6-methoxyphenyl)-5,6-dihydrocyclopenta[c]pyrazol-1(4H)-yl)pyrrolidine-1-carboxylate, and tert-butyl (3R)-3-(5-(2,3-dichloro-6-methoxyphenyl)-5,6-dihydrocyclopenta[c]pyrazol-2(4H)-yl)pyrrolidine-1-carboxylate (7 / 8 / 9):

[0164] To a solution of 5-(2,3-dichloro-6-methoxyphenyl)-1,4,5,6-tetrahydrocyclopenta[c]pyrazole (280.0 mg, 0.99 mmol) and tert-butyl (S)-3-(tolyloxy)pyrrolidine-1-carboxylate (677.3 mg, 1.98 mmol) in tetrahydrofuran (28.0 mL) was added sodium hydride (118.7 mg, 2.96 mmol, 60% in mineral oil), and the mixture was stirred at 60°C for 16 hours. After the reaction was completed, ice water was added to quench the reaction. The mixture was extracted twice with ethyl acetate, and the organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and dried in vacuo. The crude product was separated by preparative chromatography (column: Gemini-C18 150×21.2 mm, 5 μm; mobile phase: ACN-H2O (0.1% TFA), gradient: 70-75%) to give three isomers.

[0165] The first compound obtained by preparative chromatography was numbered as Example 3 (30.0 mg, yield: 7%), and its preparative chromatography RT = 6.30 min.

[0166] LC-MS: [M+H] + :452.1.

[0167] 1 H NMR (400MHz, DMSO-d6) δ7.51(d,J=9.0Hz,1H),7.18(s,1H),7.08(d,J=9.1Hz,1H),4.81(dt,J=14.3,6.9Hz,2H),3.76(s,3H),3.6 9–3.54(m,2H),3.41–3.29(m,2H),2.97(t,J=9.8Hz,2H),2.84–2.72(m,2H),2.23(tt,J=13.0,6.6Hz,2H),1.34(d,J=12.2Hz,9H).

[0168] The second compound obtained by preparative chromatography was numbered as Example 4 (40.0 mg, yield: 9%), and its preparative chromatography RT = 6.65 min.

[0169] LC-MS: [M+H] + :452.1.

[0170] 1 H NMR (400MHz, DMSO-d6) δ7.52(d,J=8.9Hz,1H),7.17(s,1H),7.08(d,J=9.0Hz,1H),4. 80(dd,J=17.6,9.0Hz,2H),3.76(d,J=2.6Hz,3H),3.67(dd,J=17.6,10.6Hz,1H),3.5 1(dd,J=11.2,4.7Hz,1H),3.43(t,J=7.5Hz,1H),3.34(dd,J=13.3,6.8Hz,1H),3.06– 2.93(m,2H),2.78(qd,J=13.7,8.9Hz,2H),2.29–2.16(m,2H),1.37(d,J=6.1Hz,9H).

[0171] The third compound obtained by preparative chromatography was numbered as Example 5 (40.0 mg, yield: 9%), and its preparative chromatography RT was 8.12 min.

[0172] LC-MS: [M+H] + :452.1.

[0173] 1 H NMR (400MHz, DMSO-d6) δ7.51(d,J=9.0Hz,1H),7.42(d,J=4.0Hz,1H),7.07(d,J=9 .0Hz,1H),4.87(d,J=5.9Hz,1H),4.60(p,J=9.1Hz,1H),3.76(s,3H),3.69(dd,J=1 0.2,4.1Hz,1H),3.54–3.43(m,2H),3.37(t,J=7.1Hz,1H),2.94(ddd,J=33.6,14. 5,8.9Hz,2H),2.78(ddd,J=14.4,9.0,5.6Hz,2H),2.33–2.23(m,2H),1.40(s,9H).

[0174] It was confirmed by two-dimensional nuclear magnetic resonance (NOESY) that Examples 3 / 4 ( FIG. 3 , FIG. 4 ) were compounds 7 / 8, and Example 5 ( FIG. 5 ) was compound 9.

[0175] Examples 6-7. 3,4-Dichloro-2-((R)-1-(R)-pyrrolidin-3-yl)-1,4,5,6-tetrahydrocyclopenta[c]pyrazol-5-yl)phenol and 3,4-Dichloro-2-((S)-1-((R)-pyrrolidin-3-yl)-1,4,5,6-tetrahydrocyclopenta[c]pyrazol-5-yl)phenol

[0176] To a solution of Example 3 (30.0 mg, 0.067 mmol) in dichloroethane (3.0 mL) at room temperature was added boron tribromide (83.3 mg, 0.33 mmol), and the mixture was stirred at room temperature for 2 hours. After completion of the reaction, the reaction was quenched with ice water (10.0 mL) and the pH was adjusted to approximately 7 with saturated sodium bicarbonate solution. The mixture was extracted twice with ethyl acetate (10.0 mL), and the combined organic phases were dried over anhydrous sodium sulfate. The solvent was concentrated under reduced pressure to dryness, and the crude product was prepared to obtain a white solid, Example 6 (3.5 mg, 16% yield).

[0177] LC-MS: [M+H] + :338.1.

[0178] 1H NMR(400MHz,DMSO-d6)δ10.40(s,1H),9.14(s,1H),7.34(d,J=8.8Hz,1H),7.2 4(s,1H),6.87(d,J=8.8Hz,1H),5.03–4.97(m,1H),4.78–4.70(m,1H),3.63–3. 53(m,2H),3.44–3.30(m,2H),3.14(dd,J=14.8,8.8Hz,1H),3.00–2.90(m,2H), 2.73(dd,J=13.6,9.1Hz,1H),2.34(dd,J=13.6,7.1Hz,1H),2.21–2.14(m,1H).

[0179] Referring to the synthesis of Example 6, Example 3 was replaced by Example 4 to obtain Example 7.

[0180] LC-MS: [M+H] + :338.1.

[0181] 1 H NMR (400MHz, DMSO-d6) δ10.37(s,1H),9.09(s,1H),7.35(d,J=8.8Hz,1H),7.25(s,1H), 6.87(d,J=8.8Hz,1H),5.02–4.96(m,1H),4.78–4.70(m,1H),3.59(dd,J=12.3,6.3Hz,1H ),3.52–3.42(m,2H),3.34(d,J=4.7Hz,1H),3.14(dd,J=14.8,8.7Hz,1H),3.01–2.91(m ,2H),2.74(dd,J=13.5,9.2Hz,1H),2.36(dd,J=13.7,7.3Hz,1H),2.20(d,J=5.1Hz,1H).

[0182] Examples 8-9. 3,4-Dichloro-2-((S)-2-(S)-pyrrolidin-3-yl)-2,4,5,6-tetrahydrocyclopenta[c]pyrazol-5-yl)phenol and 3,4-Dichloro-2-((R)-2-((S)-pyrrolidin-3-yl)-2,4,5,6-tetrahydrocyclopenta[c]pyrazol-5-yl)phenol

[0183] Synthesis of 3,4-dichloro-2-(2-((R)-pyrrolidin-3-yl)-2,4,5,6-tetrahydrocyclopenta[c]pyrazol-5-yl)phenol (2):

[0184] To a solution of tert-butyl (3R)-3-(5-(2,3-dichloro-6-methoxyphenyl)-5,6-dihydrocyclopenta[c]pyrazol-2(4H)-yl)pyrrolidine-1-carboxylate (70.0 mg, 0.155 mmol) in dichloroethane (7.0 mL) was added boron tribromide (194.4 mg, 0.78 mmol) at room temperature, and the mixture was stirred at room temperature for 3 hours. After completion of the reaction, the reaction was quenched with ice water (10.0 mL) and the pH was adjusted to approximately 7 with saturated sodium bicarbonate aqueous solution. The mixture was extracted twice with ethyl acetate (10.0 mL), and the organic phases were combined and dried over anhydrous sodium sulfate. The solvent was concentrated under reduced pressure to dryness, and the crude product was prepared to give a white solid (20.0 mg, 38% yield).

[0185] LC-MS: [M+H] + :338.1.

[0186] Preparation of 3,4-dichloro-2-((S)-2-(S)-pyrrolidin-3-yl)-2,4,5,6-tetrahydrocyclopenta[c]pyrazol-5-yl)phenol and 3,4-dichloro-2-((R)-2-((S)-pyrrolidin-3-yl)-2,4,5,6-tetrahydrocyclopenta[c]pyrazol-5-yl)phenol (3 / 4):

[0187] 3,4-Dichloro-2-(2-((R)-pyrrolidin-3-yl)-2,4,5,6-tetrahydrocyclopenta[c]pyrazol-5-yl)phenol (20.0 mg, 0.059 mmol) was chirally prepared (chromatographic column: CHIRALPAK AD-H 250 mm 20 mm, 5 μm, mobile phase: 40% EtOH (NH4OH 0.2%)) to give a pair of enantiomers.

[0188] The first compound obtained by SFC resolution was numbered as Example 8 (6.9 mg, yield 35%), which was analyzed by SFC analysis chromatographic conditions, RT = 2.92 min.

[0189] LC-MS: [M+H] + :338.0.

[0190] 1H NMR (400MHz, DMSO-d6) δ10.30(s,1H),9.07(s,2H),7.51(s,1H),7.33(d,J=8.8Hz ,1H),6.86(d,J=8.8Hz,1H),5.10(s,1H),4.56(t,J=9.2Hz,1H),3.58(d,J=5.0Hz ,2H),3.51–3.38(m,2H),3.10(dd,J=14.8,9.3Hz,1H),3.00(dd,J=14.3,9.0Hz,1 H),2.78(dt,J=14.3,9.6Hz,2H),2.41–2.31(m,1H),2.17(dd,J=8.5,4.4Hz,1H).

[0191] The second compound obtained by SFC separation was numbered as Example 9 (6.8 mg, yield 34%), which was analyzed by SFC analysis chromatographic conditions, RT = 3.88 min.

[0192] LC-MS: [M+H] + :338.0.

[0193] 1 H NMR (400MHz, DMSO-d6) δ10.32(s,1H),9.07(s,1H),7.51(s,1H),7.34(d,J=8.8H z,1H),6.86(d,J=8.8Hz,1H),5.09(s,1H),4.56(dd,J=18.4,9.2Hz,1H),3.62–3. 50(m,1H),3.49–3.36(m,2H),3.11(dd,J=14.8,9.4Hz,1H),3.00(dd,J=14.2,9. 0Hz,1H),2.82–2.73(m,2H),2.38(dd,J=13.7,7.4Hz,1H),2.27(d,J=4.3Hz,1H).

[0194] Example 10 According to the synthetic route and the preparation method of Examples 1-9, the following compound was prepared.

[0195] Example 11 Activity Test

[0196] 1. Test of compound inhibitory activity against Kv1.X

[0197] 1.1 Cell culture

[0198] HEK293 cells stably expressing Kv1.X (Kv1.1, Kv1.2, Kv1.3, Kv1.4, Kv1.5) potassium channels were cultured in DMEM medium containing 10% fetal bovine serum and 800 μg / mL G418 (Geneticin) at 37°C and a carbon dioxide concentration of 5%.

[0199] 1.2 Record the liquid used

[0200] The extracellular fluid contained 40mM NaCl, 3.5mM KCl, 1mM MgCl2·6H2O, 2mM CaCl2·2H2O, 10mM D-Glucose, 10mM HEPES, 1.25mM NaH2PO4·2H2O, and NaOH adjusted the pH to 7.4; the intracellular fluid contained 20mM KCl, 115mM K-Aspartic, 1mM MgCl2·6H2O, 5mM EGTA, 10mM HEPES, 2mM Na2-ATP, and KOH adjusted the pH to 7.2.

[0201] 1.3 Patch clamp assay

[0202] The voltage stimulation protocol for whole-cell patch-clamp recording of Kv1.X potassium currents was as follows: after whole-cell seal formation, the cell membrane voltage was clamped at -80 mV and then stepped to +40 mV for 500 ms. Data were collected repeatedly every 30 s to observe the effects of drugs on Kv1.X currents. Data were acquired using an EPC-10 amplifier (HEKA) and stored in PatchMaster (HEKA) software.

[0203] Recording electrodes are drawn from capillary glass tubes using a microelectrode puller. Under an inverted microscope, the recording electrode is placed in contact with the cell using the microelectrode manipulator. Negative pressure is applied to create a GΩ seal. After the GΩ seal is established, rapid capacitance compensation is performed. Continued negative pressure is applied to rupture the cell membrane, establishing whole-cell recording mode. Slow capacitance compensation is then performed, and membrane capacitance and series resistance are recorded. No leakage compensation is applied.

[0204] When the Kv1.X current recorded in the whole cell is stable, the drug is administered. After each drug concentration is applied for 5 minutes (or the current is stable), the next concentration is detected. Multiple concentrations are tested for each test compound. The coverslip with cells is placed in the recording bath in an inverted microscope. The test compound and the external solution without the compound are flowed through the recording bath from low concentration to high concentration by gravity perfusion to act on the cells. A vacuum pump is used for liquid exchange during the recording. The current detected in the external solution without the compound for each cell serves as its own control group. At least three cells are used for each concentration and the test is repeated three times independently. All electrophysiological experiments are performed at room temperature.

[0205] 1.4 Data Analysis

[0206] (1)Kv1.1, Kv1.2, Kv1.5-Steady state current

[0207] First, the current after each drug concentration is normalized to the blank control current. Then calculate the inhibition rate corresponding to each drug concentration

[0208] (2)Kv1.3, Kv1.4-Peak current

[0209] First, the peak current after each drug concentration was normalized with the peak current of the blank control. Then calculate the inhibition rate corresponding to each drug concentration The mean (Mean), standard deviation (SD) and standard error (SE) of the inhibition rate at each concentration were calculated. Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC 50 -X)*HillSlope))

[0210] The IC of each compound was calculated using the above equation 50 values, and a nonlinear fitting was performed for the dose-dependent effect, where IC 50 IC is the half inhibitory concentration. 50 Calculations and curve fitting were performed using GraphPad Prism software.

[0211] 2. Test of the inhibitory activity of compounds on hERG

[0212] 2.1 Cell culture

[0213] CHO cells stably expressing hERG were cultured in 35 mm diameter cell culture dishes in a 37°C, 5% CO2 incubator. The cells were passaged every 48 hours at a 1:5 ratio. The culture medium consisted of 90% F12 (Invitrogen), 10% fetal bovine serum (Gibco), 100 μg / mL G418 (Invitrogen), and 100 μg / mL Hygromycin B (Invitrogen). On the day of the experiment, the cell culture medium was aspirated, the cells were rinsed once with extracellular medium, and then digested with 0.25% Trypsin-EDTA (Invitrogen) solution at room temperature for 3-5 minutes. The digestion medium was aspirated, the cells were resuspended in extracellular medium, and then transferred to a dish for electrophysiological recording.

[0214] 2.2 Electrophysiological recording process

[0215] CHO cells stably expressing hERG potassium channels were used to record hERG potassium channel currents using the whole-cell voltage clamp technique at room temperature. Glass microelectrodes were drawn from glass electrode blanks (BF150-86-10, Sutter) using a puller. After perfusing the electrode with internal solution, the tip resistance was approximately 2-5 MΩ. The glass microelectrode was inserted into the amplifier probe to connect to the patch clamp amplifier. The clamping voltage and data recording were controlled and recorded by a computer using pClamp software, with a sampling frequency of 10 kHz and a filter frequency of 2 kHz. After obtaining the whole-cell recording, the cell was clamped at -100 mV to induce the hERG potassium current (I hERG ) was administered with a 2-second depolarization step from -100 mV to +20 mV, followed by repolarization to -50 mV for 1 second before returning to -100 mV. This voltage stimulus was administered every 5 seconds, and administration began after confirming that the hERG potassium current was stable (1 minute). Compounds were administered for at least 1 minute until steady-state or for a maximum of 3 minutes at each test concentration. At least two cells (n ≥ 2) were tested at each concentration.

[0216] 2.3 Data Processing

[0217] Data analysis and processing were performed using pClamp, GraphPad Prism 8, and Excel software. The degree of inhibition of hERG potassium current (peak hERG tail current induced at -50 mV) by different compound concentrations was calculated using the following formula: Inhibition% = [1–(I / Io)] × 100%

[0218] Wherein, Inhibition% represents the inhibition percentage of the compound on hERG potassium current, and I and Io represent the amplitudes of hERG potassium current before and after drug addition, respectively.

[0219] Compound IC 50 The results were calculated using GraphPad Prism 8 software by fitting the following equation: Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC 50 -X)*HillSlope))

[0220] Where X is the Log value of the test sample concentration, Y is the inhibition percentage at the corresponding concentration, and Bottom and Top are the minimum and maximum inhibition percentages, respectively.

[0221] 3. Experimental Results

[0222] The compounds of the present invention have inhibitory activity on Kv1.3 potassium channels, IC 50 Less than 20 nM, preferably less than 10 nM, for example, see Table 1 below.

[0223] Table 1. IC values ​​of the compounds of the present invention against Kv1.3 50 (nM) value

[0224] The compounds of the present invention have no inhibitory activity on Kv1.1, Kv1.2, Kv1.4, Kv1.5, and hERG channels. 50 All are greater than 10 μM, for example, see Table 2 below.

[0225] Table 2. IC values ​​of the compounds of the present invention for inhibition of Kv1.1, Kv1.2, Kv1.4, Kv1.5, and hERG 50 (nM) value

[0226] Therefore, the compounds of the present invention are selective Kv1.3 inhibitors, which not only have excellent activity but also excellent selectivity.

[0227] The above describes the embodiments of the present invention. However, the present invention is not limited to the above embodiments. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included in the scope of protection of the present invention.

Claims

1. A compound represented by formula (A) or its stereoisomers, or pharmaceutically acceptable salts or deuterated derivatives thereof: in, X1, X2, X3, X4, and X5 are independently selected from C and N, and two of them are N, so that A is a 5-membered heteroaryl group containing two N; R1, R2, R3, R4 are independently selected from H, F, Cl, Br, CN, C 1-4 Straight-chain or branched alkyl, C 2-4 Straight-chain or branched alkenyl, C 2-4 Straight-chain or branched alkynyl, C 1-4 Straight-chain or branched alkyloxy, C 2-4 Straight-chain or branched alkenyloxy, C 2-4 Straight-chain or branched alkynyloxy, C 3-7 Cycloalkyl. Preferably, in formula (A): R1, R2, R3, R4 are independently selected from H, F, Cl, Br, CN, C 1-4 Straight-chain or branched alkyl, C 2-4 Straight-chain or branched alkenyl, C 2-4 Straight-chain or branched-chain alkynyl. Preferably, in formula (A): R1, R2, R3, and R4 are independently selected from H, F, Cl, and Br. Preferably, in formula (A): R1 and R2 are independently selected from F, Cl, and Br, and R3 and R4 are H. Preferably, in formula (A): X1 and X4 are N, and X2, X3, and X5 are C. Preferably, in formula (A): X2 and X3 are N, and X1, X4, and X5 are C. Preferably, in formula (A): X1 and X4 are N, X2, X3, and X5 are C; R1 and R2 are independently selected from F, Cl, and Br, and R3 and R4 are H. Preferably, in formula (A): X2 and X3 are N, X1, X4, and X5 are C; R1 and R2 are independently selected from F, Cl, and Br, and R3 and R4 are H.

2. The compound according to claim 1 or its stereoisomer, or a pharmaceutically acceptable salt or deuterated product thereof, wherein the compound has the structure of formula (I): in, X1, X2, X3, X4, and X5 are independently selected from C and N, and two of them are N, so that A is a 5-membered heteroaryl group containing two N; R1, R2, R3, R4 are independently selected from H, F, Cl, Br, CN, C 1-4 Straight-chain or branched alkyl, C 2-4 Straight-chain or branched alkenyl, C 2-4 Straight-chain or branched alkynyl, C 1-4 Straight-chain or branched alkyloxy, C 2-4 Straight-chain or branched alkenyloxy, C 2-4 Straight-chain or branched alkynyloxy, C 3-7 Cycloalkyl.

3. The compound according to claim 2 or its stereoisomer, or their pharmaceutically acceptable salts or deuterated derivatives, wherein: R1, R2, R3, R4 are independently selected from H, F, Cl, Br, CN, C 1-4 Straight-chain or branched alkyl, C 2-4 Straight-chain or branched alkenyl, C 2-4 Straight-chain or branched-chain alkynyl. Preferably, R1, R2, R3, and R4 are independently selected from H, F, Cl, and Br.

4. The compound according to claim 3 or its stereoisomer, or their pharmaceutically acceptable salts or deuterated derivatives, wherein: X1 and X4 are N, X2, X3, and X5 are C; R1 and R2 are independently selected from F, Cl, and Br, and R3 and R4 are H.

5. The compound according to claim 3 or its stereoisomer, or their pharmaceutically acceptable salts or deuterated derivatives, wherein: X2 and X3 are N, X1, X4, and X5 are C; R1 and R2 are independently selected from F, Cl, and Br, and R3 and R4 are H.

6. The compound according to claim 1 or its stereoisomer, or a pharmaceutically acceptable salt or deuterated product thereof, wherein the compound has the structure of formula (II): in, X1, X2, X3, X4, and X5 are independently selected from C and N, and two of them are N, so that A is a 5-membered heteroaryl group containing two N; R1, R2, R3, R4 are independently selected from H, F, Cl, Br, CN, C 1-4 Straight-chain or branched alkyl, C 2-4 Straight-chain or branched alkenyl, C 2-4 Straight-chain or branched alkynyl, C 1-4 Straight-chain or branched alkyloxy, C 2-4 Straight-chain or branched alkenyloxy, C 2-4 Straight-chain or branched alkynyloxy, C 3-7 Cycloalkyl. Preferably, in formula (II): R1, R2, R3, R4 are independently selected from H, F, Cl, Br, CN, C 1-4 Straight-chain or branched alkyl, C 2-4 Straight-chain or branched alkenyl, C 2-4 Straight-chain or branched-chain alkynyl. Preferably, in formula (II): R1, R2, R3, and R4 are independently selected from H, F, Cl, and Br. Preferably, in formula (II): X2 and X3 are N, X1, X4, and X5 are C; R1 and R2 are independently selected from F, Cl, and Br, and R3 and R4 are H.

7. The compound according to claim 1 or its stereoisomer, or its pharmaceutically acceptable salt or deuterated substance, selected from the following compounds or their pharmaceutically acceptable salt or deuterated substance:

8. A pharmaceutical composition, characterized in that It contains the compound according to any one of claims 1 to 7 or its stereoisomer, or their pharmaceutically acceptable salts, deuterated substances, and a pharmaceutically acceptable carrier.

9. A method for preventing and / or treating Kv1.3-related diseases, characterized in that: A therapeutically and / or prophylactically effective amount of the compound according to any one of claims 1 to 7 or its stereoisomer, or their pharmaceutically acceptable salts, deuterated substances, or the pharmaceutical composition according to claim 8 is administered to an individual in need thereof. Preferably, the diseases include immune and inflammatory diseases, such as multiple sclerosis, inflammatory bowel disease, ulcerative colitis, Crohn's disease, rheumatoid arthritis, type 1 diabetes, psoriasis and asthma, spondylitis, periodontitis; as well as obesity, type 2 diabetes, renal fibrosis, Alzheimer's disease, ischemic stroke, etc.

10. Use of the compound according to any one of claims 1 to 7 or its stereoisomer, or pharmaceutically acceptable salts or deuterated substances thereof, in the preparation of a medicament for preventing and / or treating Kv1.3-related diseases. Preferably, the diseases include immune and inflammatory diseases, such as multiple sclerosis, inflammatory bowel disease, ulcerative colitis, Crohn's disease, rheumatoid arthritis, type 1 diabetes, psoriasis and asthma, spondylitis, periodontitis; as well as obesity, type 2 diabetes, renal fibrosis, Alzheimer's disease, ischemic stroke, etc.

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