Heterocyclic compound as TYK2 inhibitor and synthesis and application thereof

Abstract

The invention provides a heterocyclic compound as a TYK2 inhibitor and synthesis and application thereof. Specifically, provided is a compound or a pharmaceutically acceptable salt thereof, the compound is represented by formula A, and each group is as defined herein. # imgabs0 #

Description

Heterocyclic compounds as TYK2 inhibitors and their synthesis and application Technical Field

[0001] The present invention belongs to the fields of medicine and medical aesthetics, and specifically relates to a heterocyclic compound as a TYK2 inhibitor, and its synthesis and application. Background Art

[0002] Janus kinases (JAKs) are intracellular, non-receptor tyrosine kinases that mediate signaling and activation of various cytokines. The JAK kinase family is divided into four subtypes: JAK1, JAK2, JAK3, and TYK2. Each subtype mediates a different type of cytokine signaling pathway. JAK1, JAK2, and TYK2 are expressed in various tissues and cells throughout the body, while JAK3 is primarily expressed in hematopoietic tissues. A common feature of cytokine receptors is that the receptors themselves lack kinase activity, but their intracellular domains contain binding sites for the tyrosine kinases JAK. When cytokine receptors bind to their ligands, they activate the receptor-associated JAKs, leading to receptor phosphorylation. The phosphorylated tyrosine sites can bind to STAT proteins containing SH2 domains, leading to STAT recruitment to the receptors and phosphorylation by JAKs. Subsequently, phosphotyrosine-mediated STAT dimerization occurs, and the activated STAT dimers translocate to the nucleus and activate the transcription of their target genes, thereby regulating various cell functions, including growth, activation, and differentiation.

[0003] TYK2 is the first subtype of the JAK family to be discovered. It mediates the functions of cytokines such as IFN-a, IL-6, IL-10, IL-12, and IL-23. Studies have shown that TYK2 deletion mutations can effectively inhibit the occurrence of immune diseases such as allergies, autoimmunity, and inflammation. IL-23 plays a crucial role in the development and progression of psoriasis. Recent studies have shown that the pathogenesis of psoriasis is that endogenous unknown antigens activate antigen-presenting cells (APCs) to secrete IL-23. IL-23 activates Th17 cells to secrete cytokines such as IL-17, inducing keratinocyte differentiation and division and secretion of IL-23, further stimulating inflammation and keratinocyte proliferation to produce psoriasis. TYK2 and JAK2 jointly mediate the downstream signaling pathways of IL-23. Inhibiting JAK2 can lead to anemia and other blood-related side effects. Therefore, targeting TYK2 is a good strategy to inhibit the IL-23 signaling pathway.

[0004] Early TYK2 inhibitors such as Tofacitinib are all non-selective JAK inhibitors. They are the first oral JAK inhibitors with significant inhibitory activity against JAK1, 2, and 3 subtypes. The inhibition of the activity of other subtypes such as JAK1, JAK2, and JAK3 increases the efficacy of tofacitinib, but it also brings more serious side effects, including infection, tuberculosis, tumors, anemia, liver damage, and increased cholesterol. Because JAK2 activity is related to erythroid cell differentiation and lipid metabolism, some of the above-mentioned adverse reactions such as anemia are believed to be related to the insufficient selectivity of Tofacitinib for JAK2 and are caused by the non-selective inhibition of the drug. There are currently no TYK2 selective inhibitors on the market. Early JAK inhibitors mainly work by competing for the binding of the kinase domain to ATP, so there is a general problem of low selectivity.

[0005] TYK2 is also implicated in some cancers. For example, abnormal survival of acute lymphoblastic leukemia (T-ALL) cells is associated with TYK2 activation. Gene knockout experiments have shown that 88% of T-ALL cell lines and 63% of patient-derived T-ALL cells are dependent on TYK2, thus implicating TYK2 as an oncogene in T-ALL. The TYK2-selective inhibitor NDI-031301 can inhibit the growth of human T-ALL cell lines by inducing apoptosis. It also demonstrates good safety and efficacy in a mouse model bearing KOPT-K1 T-ALL tumor cells, demonstrating the promise of TYK2-selective inhibitors in the treatment of T-ALL.

[0006] Given the good efficacy of non-selective JAK inhibitors and the serious side effects associated with multiple targets, a safer TYK2 selective inhibitor is being developed for psoriasis, lupus erythematosus, inflammatory bowel disease, psoriatic arthritis, arthritis, vasculitis, fibrosis, dermatitis, skin aging, encephalitis, lupus nephritis, neuroinflammation, different types of multiple sclerosis (including optic neuritis, neuromyelitis optica), chronic inflammatory demyelinating polyneuropathy, Parkinson's disease, dementia, amyotrophic lateral sclerosis, myasthenia gravis, mental illness, schizophrenia, epilepsy, spinal injury, sleep deprivation, etc. It has great clinical application potential in the treatment of various autoimmune and inflammatory diseases related to TYK2, including insomnia, brain injury, stroke, neuropsychiatric lupus, diabetic encephalopathy, sepsis-related encephalopathy, central nervous system tumors, Huntington's disease, post-operative neurological syndrome, pain, itching, depression, narcolepsy, hydrocephalus, ankylosing spondylitis, respiratory diseases, diabetes, inflammatory eye diseases, hepatitis, cardiovascular diseases, systemic sclerosis, organ transplantation, alopecia areata, acne, eczema, vitiligo, Sjögren's syndrome, viral inflammation, and some cancers.

[0007] Therefore, there is an urgent need in the art to develop novel brain-penetrating TYK2 inhibitors with high selectivity and better efficacy, especially brain-penetrating TYK2 inhibitors suitable for central nervous system diseases.

[0008] Summary of the Invention

[0009] The purpose of the present invention is to provide a class of brain-penetrating TYK2 inhibitors with novel structure, high safety and excellent efficacy.

[0010] In the first aspect of the present invention, a compound or a pharmaceutically acceptable salt thereof is provided, wherein the compound is as shown in Formula A1,

[0011] in,

[0012] X is N or CR8; R8 is selected from the group consisting of H, optionally substituted C 1-4 alkyl;

[0013] Y, R1, R2, R3, R4, R5, R6, R7, R9 and "optionally substituted" are as defined in Formula A.

[0014] In another preferred embodiment, a compound or a pharmaceutically acceptable salt thereof is provided, wherein the compound is as shown in Formula A,

[0015] Y is N or CR9;

[0016] R1 is selected from the group consisting of H, optionally substituted C 1-4 alkyl;

[0017] R2 is selected from the group consisting of H, optionally substituted C 1-4 alkyl;

[0018] R3 and R4 are each independently selected from the group consisting of H, halogen, optionally substituted C 1-4 alkyl;

[0019] R5 is selected from the group consisting of H, optionally substituted C 1-6 alkyl;

[0020] R6 is selected from the group consisting of H, optionally substituted C 1-4 alkyl;

[0021] R7 is selected from the group consisting of H, optionally substituted C 1-4 alkyl;

[0022] R9 is selected from the group consisting of H, optionally substituted C 1-4 alkyl;

[0023] The term "optionally substituted" means that the group is unsubstituted or substituted with one or more substituents selected from the group consisting of deuterium, halogen, C1-4 Alkyl, C 1-4 Halogenated alkyl.

[0024] In another preferred embodiment, R4 is selected from the following group: H, methyl, ethyl.

[0025] In another preferred embodiment, R4 is H.

[0026] In another preferred embodiment, R5 is C 1-6 alkyl.

[0027] In another preferred embodiment, R5 is selected from the following group: methyl, ethyl, n-propyl, and n-butyl.

[0028] In another preferred embodiment, R5 is ethyl.

[0029] In another preferred embodiment, R6 is selected from the following group: H, methyl, ethyl.

[0030] In another preferred embodiment, R6 is H.

[0031] In another preferred embodiment, R6 is selected from the following group: H, methyl, ethyl.

[0032] In another preferred embodiment, R6 is H.

[0033] In another preferred embodiment, R7 is selected from the following group: H, methyl, ethyl.

[0034] In another preferred embodiment, R7 is H.

[0035] In another preferred embodiment, the compound is as shown in formula A1-S or formula A1-R

[0036] In another preferred embodiment, the compound is as shown in formula AS or formula AR

[0037] In another preferred embodiment, the compound is as shown in formula AS (i.e., S configuration). In another preferred embodiment, the compound is as shown in formula I1

[0038] In another preferred embodiment, the compound is as shown in Formula I

[0039] In another preferred embodiment, R8 and R9 are each independently selected from the following group: H, methyl, ethyl.

[0040] In another preferred embodiment, R8 is H.

[0041] In another preferred embodiment, R9 is H.

[0042] In another preferred embodiment, X is N or CH.

[0043] In another preferred embodiment, X is CR8.

[0044] In another preferred embodiment, X is CH.

[0045] In another preferred embodiment, Y is N or CH.

[0046] In another preferred embodiment, Y is N.

[0047] In another preferred embodiment, R1 is selected from the following group: C 1-4 Alkyl, deuterated C 1-4 alkyl.

[0048] In another preferred embodiment, R1 is selected from the following group: methyl, ethyl, deuterated methyl, deuterated ethyl.

[0049] In another preferred embodiment, R1 is selected from the following group: -CH3, -CD3.

[0050] In another preferred embodiment, R2 is selected from the following group: C 1-4 Alkyl, deuterated C 1-4 alkyl.

[0051] In another preferred embodiment, R2 is selected from the group consisting of methyl, ethyl, deuterated methyl, and deuterated ethyl.

[0052] In another preferred embodiment, R2 is selected from the following group: -CH3, -CD3.

[0053] In another preferred embodiment, R3 is selected from the following group: H, C 1-4 alkyl.

[0054] In another preferred embodiment, R3 is selected from the following group: H, methyl, ethyl.

[0055] In another preferred embodiment, the compound is as shown in formula I1-S or formula I1-R

[0056] In another preferred embodiment, the compound is as shown in Formula IS or Formula IR

[0057] In another preferred embodiment, the compound is as shown in formula IS (ie, S configuration).

[0058] In another preferred embodiment, X, Y, R1, R2, R3, R4, R5, R6, R7, R8 and R9 are each independently a corresponding group of a specific compound shown in the Examples, Table A or Table B.

[0059] In another preferred embodiment, the compound is a compound selected from Table A or Table B or a pharmaceutically acceptable salt thereof,

[0060] Table A

[0061] Table B

[0062] In another preferred embodiment, the compound is compound 1S, 2S, 3S, 4S, 5S, or 6S in Table B.

[0063] In another preferred embodiment, the compound is compound 4S in Table B.

[0064] In a second aspect of the present invention, there is provided a pharmaceutical composition comprising:

[0065] (i) the compound according to the first aspect or a pharmaceutically acceptable salt thereof, and (ii) a pharmaceutically acceptable carrier or excipient.

[0066] In a third aspect of the present invention, there is provided a use of the compound according to the first aspect or the pharmaceutical composition according to the second aspect in the preparation of (i) a medicament for treating or preventing a TYK2-mediated disease and / or (ii) a TYK2 inhibitor.

[0067] In another preferred embodiment, the TYK2-mediated diseases include: psoriasis, lupus erythematosus, inflammatory bowel disease, psoriatic arthritis, arthritis, vasculitis, fibrosis, dermatitis, skin aging, encephalitis, lupus nephritis, neuroinflammation, multiple sclerosis (including optic neuritis and neuromyelitis optica), chronic inflammatory demyelinating polyneuropathy, Parkinson's disease, dementia, amyotrophic lateral sclerosis, myasthenia gravis, mental illness, schizophrenia, epilepsy, spinal injury, sleep disorders, brain injury, stroke, neuropsychiatric lupus, diabetic encephalopathy, sepsis-related encephalopathy, central nervous system tumors, Huntington's disease, postoperative neurological syndrome, pain, itching, depression, narcolepsy, hydrocephalus, ankylosing spondylitis, respiratory diseases, diabetes, inflammatory eye diseases, hepatitis, cardiovascular diseases, systemic sclerosis, organ transplantation, alopecia areata, acne, eczema, vitiligo, Sjögren's syndrome, viral inflammation, cancer, or a combination thereof.

[0068] In a fourth aspect of the present invention, a method for inhibiting TYK2 is provided, comprising:

[0069] The subject is contacted with the compound of the first aspect, thereby inhibiting TYK2 activity in the subject.

[0070] In another preferred embodiment, the object is a cell.

[0071] In another preferred embodiment, the method is non-therapeutic in vitro.

[0072] In a fifth aspect of the present invention, a method for inhibiting the expression of pSTAT5 in a cell is provided, the method comprising:

[0073] The subject is contacted with the compound according to the first aspect, thereby inhibiting the expression of pSTAT5 in the cell.

[0074] In another preferred embodiment, the method is non-therapeutic in vitro.

[0075] In a sixth aspect of the present invention, a method for treating or preventing a TYK2-mediated disease is provided, comprising the steps of:

[0076] A safe and effective amount of the compound of the first aspect or the pharmaceutical composition of the second aspect is administered to a person in need thereof to treat or prevent a TYK2-mediated disease.

[0077] In another preferred embodiment, the TYK2-mediated diseases include: psoriasis, lupus erythematosus, inflammatory bowel disease, psoriatic arthritis, arthritis, vasculitis, fibrosis, dermatitis, skin aging, encephalitis, lupus nephritis, neuroinflammation, multiple sclerosis (including optic neuritis and neuromyelitis optica), chronic inflammatory demyelinating polyneuropathy, Parkinson's disease, dementia, amyotrophic lateral sclerosis, myasthenia gravis, mental illness, schizophrenia, epilepsy, spinal injury, sleep disorders, brain injury, stroke, neuropsychiatric lupus, diabetic encephalopathy, sepsis-related encephalopathy, central nervous system tumors, Huntington's disease, postoperative neurological syndrome, pain, itching, depression, narcolepsy, hydrocephalus, ankylosing spondylitis, respiratory diseases, diabetes, inflammatory eye diseases, hepatitis, cardiovascular diseases, systemic sclerosis, organ transplantation, alopecia areata, acne, eczema, vitiligo, Sjögren's syndrome, viral inflammation, cancer, or a combination thereof.

[0078] In a seventh aspect of the present invention, a method for preparing a compound is provided, wherein the compound is a compound as shown in Formula IS,

[0079] And the preparation method comprises the steps of:

[0080] Reacting the intermediate of formula II-S with the intermediate of formula III to obtain a compound of formula IS;

[0081] In each formula, Y, R1, R2 and R3 are as defined in Formula I.

[0082] In the eighth aspect of the present invention, there is provided an intermediate of formula II-S,

[0083] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one. BRIEF DESCRIPTION OF THE DRAWINGS

[0084] FIG1A and FIG1B show the animal body weight and clinical score results in Test Example 3. FIG.

[0085] 2A and 2B show the results of the levels of TNF-α and IFN-γ in serum in Test Example 3. DETAILED DESCRIPTION

[0086] After extensive and in-depth research, the inventor unexpectedly discovered a class of novel structures (such as ), especially the S configuration of the compound, has excellent TYK2 inhibitory activity and lower toxicity. In addition, the inventors have also overcome the difficulties in synthesizing chiral The difficulty of structural compound identification is solved, thereby avoiding the use of chiral chromatography, thereby providing a class of compounds that are easier to obtain industrially, have excellent TYK2 inhibitory activity and lower toxicity. Based on this, the inventors completed the present invention.

[0087] the term

[0088] As used herein, the term "halogen" refers to F, Cl, Br, or I. Correspondingly, "halo" refers to a group in which a hydrogen atom is replaced by F, Cl, Br, or I.

[0089] Unless otherwise indicated, the term "alkyl" by itself or as part of another substituent refers to a straight or branched chain hydrocarbon radical (i.e., C 1-6 = represents 1-6 carbons). Preferably, the alkyl group has 1-4 carbons, i.e., C1-4 alkyl, and more preferably has 1-3 carbons, i.e., C 1-3 Alkyl. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, and the like.

[0090] As used herein, the terms "comprising", "including" or "comprising" indicate that various components can be used together in the mixture or composition of the present invention. Therefore, the terms "consisting essentially of" and "consisting of" are included in the term "comprising".

[0091] As used herein, the term "deuterated" refers to a group in which one or more hydrogen atoms are replaced by deuterium. Preferably, it refers to a group in which all hydrogen atoms are replaced by deuterium.

[0092] As used herein, the term "pharmaceutically acceptable" refers to substances that are suitable for use in humans and / or animals without excessive adverse side effects (such as toxicity, irritation, and allergic response), ie, at a reasonable benefit / risk ratio.

[0093] Unless otherwise specified, all compounds mentioned in the present invention are intended to include all possible optical isomers, such as single chiral compounds or mixtures of various chiral compounds (i.e., racemates). In all compounds of the present invention, each chiral carbon atom may optionally be in the R configuration or the S configuration, or a mixture of the R and S configurations.

[0094] Certain compounds of the present invention possess asymmetric carbon atoms (optical centers) or double bonds; racemates, diastereomers, geometric isomers, regioisomers, and individual isomers (e.g., isolated enantiomers) are intended to be encompassed within the scope of the present invention. When compounds provided herein have defined stereochemistry (denoted as R or S, or indicated by dashed or wedge-shaped bonds), those skilled in the art will understand that those compounds are substantially free of other isomers (e.g., at least 80%, 90%, 95%, 98%, 99%, and up to 100% free of other isomers).

[0095] The compounds of the present invention may also contain unnatural proportions of atomic isotopes at one or more of the isotopic atoms that make up such compounds. An unnatural proportion of an isotope can be defined as the amount of the atom in question found in nature to 100% of that atom. For example, a compound may incorporate a radioactive isotope, such as tritium ( 3 H), iodine-125 ( 125 I) or carbon-14 ( 14 C), or non-radioactive isotopes such as deuterium ( 2 H) or carbon-13 ( 13 C) In addition to those uses described herein, such isotopic variants may provide additional uses. For example, isotopic variants of the compounds of the present invention may have additional uses, including but not limited to, as diagnostic and / or imaging agents, or as cytotoxic / radiotoxic therapeutic agents. Furthermore, isotopic variants of the compounds of the present invention may have altered pharmacokinetic and pharmacodynamic properties, thereby contributing to increased safety, tolerability, or efficacy during treatment. All isotopic variants of the compounds of the present invention, whether or not radioactive, are intended to be encompassed by the present invention.

[0096] Active ingredient

[0097] As used herein, the term "compound of the present invention" or "compound of the present invention" refers to a compound of formula (A) or formula (I). The term also includes various crystalline forms, pharmaceutically acceptable salts, hydrates, or solvates of the compound of formula (A) or formula (I).

[0098] The term "pharmaceutically acceptable salt" refers to a salt formed by a compound of the present invention with an acid or base that is suitable for use as a pharmaceutical. Pharmaceutically acceptable salts include inorganic salts and organic salts. A preferred class of salts is a salt formed by a compound of the present invention with an acid. Suitable acids for forming salts include, but are not limited to, inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, and phosphoric acid; organic acids such as formic acid, acetic acid, trifluoroacetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, benzoic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, and naphthalenesulfonic acid; and amino acids such as proline, phenylalanine, aspartic acid, and glutamic acid. Another preferred salt is a salt of the compound of the present invention formed with a base, such as an alkali metal salt (e.g., sodium salt or potassium salt), an alkaline earth metal salt (e.g., magnesium salt or calcium salt), an ammonium salt (e.g., lower alkanolammonium salt and other pharmaceutically acceptable amine salts), for example, methylamine salt, ethylamine salt, propylamine salt, dimethylamine salt, trimethylamine salt, diethylamine salt, triethylamine salt, tert-butylamine salt, ethylenediamine salt, hydroxyethylamine salt, dihydroxyethylamine salt, trihydroxyethylamine salt, and amine salts formed from morpholine, piperazine, and lysine, respectively.

[0099] The term "solvate" refers to a complex formed by the coordination of a compound of the present invention with solvent molecules in a specific ratio. "Hydrate" refers to a complex formed by the coordination of a compound of the present invention with water.

[0100] In addition, the compounds of the present invention also include prodrugs of the compounds represented by Formula (A) or Formula (I). The term "prodrug" includes a compound that may be biologically active or inactive, and when administered by an appropriate method, undergoes metabolism or chemical reactions in the human body to transform into a compound of Formula (A) or Formula (I), or a salt or solution composed of a compound of Formula (A) or Formula (I). The prodrugs include (but are not limited to) carboxylate, carbonate, phosphate, nitrate, sulfate, sulfone, sulfoxide, amino, carbamate, azo, phosphoramide, glucoside, ether, acetal, and the like forms of the compounds.

[0101] Preparation method

[0102] The rest of this document describes in more detail methods for preparing compounds of formula (A) or (I) of the present invention, but these specific methods do not limit the present invention in any way. The compounds of the present invention can also be conveniently prepared by optionally combining various synthetic methods described in this specification or known in the art, and such combinations can be readily performed by those skilled in the art.

[0103] Pharmaceutical compositions and methods of administration

[0104] Since the compounds of the present invention have excellent inhibitory activity against TYK2 (selective inhibitory activity), the compounds of the present invention and their various crystalline forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and pharmaceutical compositions containing the compounds of the present invention as the main active ingredient can be used to treat, prevent and alleviate diseases mediated by TYK2 (i.e., TYK2-mediated diseases). According to the prior art, the compounds of the present invention can be used to treat the following diseases: psoriasis, lupus erythematosus, inflammatory bowel disease, psoriatic arthritis, arthritis, vasculitis, fibrosis, dermatitis, skin aging, encephalitis, lupus nephritis, neuroinflammation, different types of multiple sclerosis (including optic neuritis, neuromyelitis optica), chronic inflammatory demyelinating polyneuropathy, Parkinson's disease, dementia, amyotrophic lateral sclerosis, myasthenia gravis, mental illness, schizophrenia, epilepsy, spinal injury, sleep disorders, brain injury, stroke, neuropsychiatric lupus, diabetic encephalopathy, sepsis-related encephalopathy, central nervous system tumors, Huntington's disease, postoperative neurological syndrome, pain, itching, depression, narcolepsy, hydrocephalus, ankylosing spondylitis, respiratory diseases, diabetes, inflammatory eye diseases, hepatitis, cardiovascular diseases, systemic sclerosis, organ transplantation, alopecia areata, acne, eczema, vitiligo, Sjögren's syndrome, viral inflammation, some cancers, etc.

[0105] The pharmaceutical composition of the present invention comprises a safe and effective amount of a compound of the present invention or a pharmacologically acceptable salt thereof, and a pharmacologically acceptable excipient or carrier. "Safe and effective amount" means an amount of the compound sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 1-500 mg of the compound of the present invention per dose, more preferably 1-200 mg per dose. Preferably, "one dose" is one capsule or tablet.

[0106] "Pharmaceutically acceptable carriers" refer to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as Tween ), wetting agents (such as sodium lauryl sulfate), colorants, flavorings, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0107] There is no particular limitation on the administration of the compound or pharmaceutical composition of the present invention. Representative administration routes include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

[0108] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0109] Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifying agents, and the release of the active compound or compounds in such compositions can be delayed in a certain portion of the digestive tract. Examples of useful encapsulating components are polymeric substances and waxes. If desired, the active compound can also be microencapsulated with one or more of the above-mentioned excipients.

[0110] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, or mixtures thereof.

[0111] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0112] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0113] Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0114] Dosage forms for topical administration of the compounds of this invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required.

[0115] The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable compounds.

[0116] When using a pharmaceutical composition, a safe and effective amount of the compound of the present invention is administered to a mammal (e.g., a human) in need of treatment, wherein the dosage is a pharmaceutically effective dosage. For a 60 kg human, the daily dosage is generally 1 to 500 mg, preferably 1 to 200 mg. Of course, the specific dosage will also take into account factors such as the route of administration and the patient's health condition, all of which are within the skill of a skilled physician.

[0117] The main advantages of the present invention include:

[0118] 1. The compounds of the present invention (especially the compounds of the present invention in S configuration) have excellent inhibitory activity against TYK2.

[0119] 2. The compounds of the present invention have excellent inhibitory effects on pSTAT5 expression.

[0120] 3. The compounds of the present invention have excellent brain penetrance (ability to penetrate the blood-brain barrier).

[0121] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention and are not intended to limit the scope of the invention. The experimental methods in the following examples, for which no specific conditions are specified, are generally based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are by weight.

[0122] Preparation Example

[0123] Example 1

[0124] Step A

[0125] Ethylmagnesium bromide (24 ml, 71.6 mmol) was added to a three-necked flask containing 200 ml of tetrahydrofuran at 0°C. A solution of compound 1a (6 g, 34.1 mmol) in tetrahydrofuran was slowly added dropwise under nitrogen at 0°C and stirred at room temperature for 5 h. After completion, the reaction was quenched with saturated ammonium chloride solution (100 ml), diluted with water (100 mL), extracted with ethyl acetate (100 mL x 3), washed with saturated sodium chloride solution (100 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was collected and concentrated under reduced pressure to yield a yellow liquid. This was separated by column chromatography (petroleum ether / ethyl acetate = 4 / 1) to afford compound 1b (3.55 g, 50.57%) as a yellow liquid. LCMS: 206.0, 208.0 [M+H].

[0126] Step B

[0127] Compound 1b (3.55 g, 17.2 mmol) and Dess-Martin periodinane (14.59 g, 34.4 mmol) were added sequentially to a 250 ml round-bottom flask containing 100 ml of anhydrous dichloromethane and stirred at room temperature for 16 hours. After completion of the reaction, saturated aqueous sodium bicarbonate solution was added to adjust the pH to 7-8, as monitored by LCMS. The mixture was extracted with dichloromethane (100 ml x 3). The combined extracts were washed with saturated brine (100 ml). The organic phase was collected, dried over anhydrous Na2SO4, concentrated under reduced pressure, and separated by column chromatography (petroleum ether / ethyl acetate, 4 / 1) to afford compound 1c (3.2 g, 91.16%) as a yellow solid. LCMS: 204.0, 206.0 [M+H]+.

[0128] Step C

[0129] Compound 1c (300 mg, 1.47 mmol) and compound 1d (245.1 mg, 2.21 mmol) were dissolved in dioxane (10 mL). Pd2(dba)3 (238.35 mg, 0.29 mmol), cesium carbonate (1.44 g, 4.41 mmol), and 1,1'-bis(dicyclohexylphosphino)ferrocene (dcpf, 335.5 mg, 0.59 mmol) were added sequentially. The mixture was stirred at 80°C for 3 hours under nitrogen atmosphere. LCMS monitoring was performed. After completion of the reaction, the mixture was concentrated under reduced pressure, diluted with water (20 mL), and extracted with ethyl acetate (20 mL x 3). The combined extracts were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography (petroleum ether / ethyl acetate, 3 / 1) to afford compound 1e (289 mg, 70.56%) as a yellow solid. 1HNMR (400MHz, CD3OD) δ8.59(s,1H),8.33(s,1H),3.04(q,J=7.2Hz,2H),2.27(dd,J=7.4,4.2Hz,1H),1.58(t,J=3.9Hz,1 H), 1.47 (dd, J=7.4, 3.6Hz, 1H), 1.19 (t, J=7.2Hz, 3H), 1.04-0.94 (m, 3H), 0.92-0.87 (m, 1H). LCMS: 279.1, 281.1[M+H]+.

[0130] Step D

[0131] Compound 1f (800 mg, 3.94 mmol) and compound 1g (500 mg, 3.94 mmol) were dissolved in a dioxane / water mixture (20 mL / 4 mL). Pd(dppf)Cl2-CH2Cl2 (160 mg, 0.20 mmol) and potassium carbonate (1.09 g, 7.88 mmol) were added sequentially. The mixture was stirred at 110°C under nitrogen for 16 hours. The reaction was monitored by LCMS. After completion, the mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL x 3). The combined extracts were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by medium-pressure flash chromatography (dichloromethane / methanol, 15 / 1) to afford compound 1h (550 mg, 68.06%) as an off-white solid. 1HNMR (400MHz, DMSO-d6): δppm: 8.16 (s, 1H), 7.73 (d, J = 5.2Hz, 1H), 6.96 (d, J = 5.6Hz, 1H), 6.07 (s, 2H), 4.23 (s, 3H), 3.64 (s, 3H). LCMS: 205.9 [M+H]+.

[0132] Step E

[0133] Compound 1e (75 mg, 0.27 mmol) and compound 1h (55.2 mg, 0.27 mmol) were dissolved in dioxane (5 mL), and XantPhos (62.28 mg, 0.11 mmol), cesium carbonate (263 mg, 0.81 mmol), and Pd2(dba)3 (49.2 mg, 0.054 mmol) were added sequentially. The mixture was stirred in a microwave oven at 115°C for 2 hours under a nitrogen atmosphere. LCMS monitoring was performed. After completion of the reaction, the mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL × 3). The combined extracts were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by medium-pressure flash chromatography (dichloromethane / methanol, 20 / 1) to yield 200 mg of the crude product. The crude product was separated by preparative HPLC (acetonitrile / water = 55:45; Gemini 5uC18 150x21.2 mm) to obtain 25 mg of product, which was then separated by SFC (Thar preparative 80, column: CHIRALPAK IC 250 mmx20 mmx5 μm, Modifier: 40% methanol (NH4OH 0.2%) / 60% CO2, total flow rate: 40 g / min, temperature: 40°C) to obtain two isomers: white solid compound 1R (6.1 mg, Rt: 3.65 min) and white solid compound 1S (7.4 mg, Rt: 7.82 min) with a total yield of 11.26%.

[0134] Compound 1R: 1HNMR (400MHz, DMSO) δ12.33 (s, 1H), 10.74 (s, 1H), 9.69 (s, 1H), 8.95 (s, 1H), 8.33 (s, 1H), 8.17 (d, J = 5.2 Hz, 1H), 7.47 (d, J = 5.2 Hz, 1H), 4.28 (s, 3H), 3.81 (s, 3H), 3.17 (q, J=7.3Hz,2H),2.42(dd,J=7.4,4.4Hz,1H),1.44(t,J=3.8Hz,1H),1.35(dd,J=7.4,3.2Hz, 1H), 1.14 (t, J=7.2Hz, 3H), 0.95-0.82 (m, 3H), 0.78 (d, J=5.4Hz, 1H). LCMS: 448.2[M+H]+.

[0135] Compound 1S: 1HNMR (400MHz, DMSO) δ12.33 (s, 1H), 10.74 (s, 1H), 9.69 (s, 1H), 8.95 (s, 1H), 8.33 (s, 1H), 8.17 (d, J = 5.2 Hz, 1H), 7.47 (d, J = 5.2 Hz, 1H), 4.28 (s, 3H), 3.81 (s, 3H), 3.17 (q, J=7.3Hz,2H),2.42(dd,J=7.4,4.4Hz,1H),1.44(t,J=3.8Hz,1H),1.35(dd,J=7.4,3.2Hz, 1H), 1.14 (t, J=7.2Hz, 3H), 0.95-0.82 (m, 3H), 0.78 (d, J=5.4Hz, 1H). LCMS: 448.2[M+H]+.

[0136] Example 2

[0137] Step A

[0138] Compound 2a (1.3 g, 6.4 mmol) was dissolved in dioxane (35 mL), and pinacol diboron (4.08 g, 16.08 mmol), potassium carbonate (0.63 g, 4.56 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride dichloromethane complex (0.47 g, 0.64 mmol) were added sequentially. The mixture was stirred at 90°C for 5 hours under nitrogen atmosphere. After completion of the reaction, the mixture was cooled to room temperature and concentrated under reduced pressure to obtain the crude product, which was then separated by medium-pressure flash chromatography (petroleum ether / ethyl acetate, 2 / 1) to afford Compound 2b (1 g, 43.4%) as a yellow solid. 1HNMR (400MHz, DMSO) δ6.82-6.77 (m, 2H), 6.75 (d, J = 4.4Hz, 1H), 4.82 (s, 2H), 3.63 (s, 3H), 1.28 (s, 12H). LCMS: 250.1[M+H]+.

[0139] Step B

[0140] Compound 2b (1.04 g, 4.17 mmol) was dissolved in dioxane (16 mL) and water (4 mL). Compound 2c (450 mg, 2.78 mmol), tetrakis(triphenylphosphine)palladium (160.51 mg, 0.139 mmol), and cesium carbonate (1.82 g, 5.56 mmol) were added sequentially. The mixture was stirred at 90°C under nitrogen for 6 hours. LCMS monitoring was used. After completion of the reaction, the mixture was cooled to room temperature and concentrated under reduced pressure. The residue was dissolved in ethyl acetate (100 mL) and washed with water (50 mL) and saturated brine (50 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product. The crude product was separated by medium-pressure flash chromatography (petroleum ether / ethyl acetate, 5 / 1) to afford Compound 2d (0.55 g, 63.9%) as a yellow solid. 1HNMR (400MHz, DMSO-d6): δppm8.47(s,1H),6.96-6.94(m,1H),6.86(t,J=8.0Hz,1H),6.75-6.73(m,1H),4.98(s,2H),3.91(s,3H),3.66(s,3H). LCMS:205.1[M+H]+.

[0141] Step C

[0142] Compound 2e (1 g, 5.2 mmol) was added to a single-necked flask containing 15 mL of N,N-dimethylformamide at room temperature. Compound 2f (0.61 g, 6.24 mmol), N,N-diisopropylethylamine (2.01 g, 15.6 mmol), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (2.96 g, 7.8 mmol) were added with stirring at room temperature. The mixture was stirred at room temperature for 16 h. LCMS monitoring was performed. After completion of the reaction, the reaction solution was poured into 50 mL of ice water and extracted with ethyl acetate (50 mL x 3). The ethyl acetate phase was washed with saturated sodium chloride (20 mL x 3), dried over anhydrous Na2SO4, filtered, and the filtrate was collected and concentrated under reduced pressure to obtain the crude product as a yellow liquid. The crude product was separated by column chromatography (ethyl acetate / petroleum ether, 1 / 5) to obtain 2 g of compound 2 (1.1 g, 80.77%) as a yellow solid. LCMS: 235.0, 237.0 [M+H]+.

[0143] Step D

[0144] Compound 2g (460 mg, 1.96 mmol) and compound 2d (400 mg, 1.96 mmol) were dissolved in dry N,N-dimethylformamide (10 mL). Sodium hydride (234 mg, 5.88 mmol; 60%) was added at 0°C under nitrogen and stirred at room temperature for 16 hours. LCMS monitoring was used. After completion of the reaction, the mixture was quenched with water (20 mL) and extracted with ethyl acetate (50 mL × 2). The combined extracts were washed with saturated brine (30 mL × 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product. The crude product was separated by medium-pressure flash chromatography (petroleum ether / ethyl acetate, 3 / 1) to afford compound 2h (500 mg, 66.5%) as a yellow solid. LCMS: 403.0 [M+H]+.

[0145] Step E

[0146] Ethylmagnesium bromide (0.7 ml, 2 mmol) was added to a three-necked flask containing 200 ml of tetrahydrofuran at 0°C. A solution of compound 2h (500 mg, 1.2 mmol) in tetrahydrofuran was slowly added dropwise under nitrogen at 0°C and stirred at room temperature for 5 h. LCMS monitoring was used. After completion of the reaction, the reaction was quenched with saturated ammonium chloride solution (100 ml), diluted with water (100 mL), extracted with ethyl acetate (100 mL x 3), washed with saturated sodium chloride solution (100 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was collected and concentrated under reduced pressure to obtain a yellow liquid. This was separated by column chromatography (petroleum ether / ethyl acetate, 4 / 1) to obtain a yellow liquid compound 2i (400 mg, 86.2%).

[0147] Step F

[0148] Compound 2i (50 mg, 0.2 mmol) and compound 1d (22.4 mg, 0.2 mmol) were dissolved in dry dioxane (3 mL). 4,5-bis(diphenylphosphino)-9,9-dimethoxyanthracene (15.6 mg, 0.0269 mmol), cesium carbonate (88.2 mg, 0.269 mmol), and tris(dibenzylideneacetone)dipalladium (10.9 mg, 0.0135 mmol) were added sequentially. The mixture was stirred in a microwave oven at 115°C for 1 hour under a nitrogen atmosphere. LCMS monitoring was performed. After completion of the reaction, the mixture was cooled to room temperature and concentrated under reduced pressure. The residue was dissolved in ethyl acetate (50 mL) and washed with water (30 mL) and saturated brine (30 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product, which was separated by medium-pressure flash chromatography (dichloromethane / methanol, 10 / 1 to 5 / 1) to yield 40 mg of the crude product. The crude product was separated by preparative HPLC (acetonitrile / water, 55 / 45; Gemini 5uC18 150x21.2 mm) to obtain 20 mg of the product, which was then separated by SFC (Thar Prep 80, column: CHIRALPAKAD-H 250mmx20mmx5μm, Modifier: 40% methanol (NH4OH 0.2%) / 60% CO2, total flow rate: 40 g / min, temperature: 40°C) to obtain two isomers: compound 2R (3.6 mg, Rt: 2.71 min) and compound 2S (3.9 mg, Rt: 3.51 min) with a total yield of 12.3%.

[0149] Compound 2R: 1HNMR (400MHz, DMSO) δ11.09 (s, 1H), 10.74 (s, 1H), 8.88 (s, 1H), 8.13 (s, 1H), 8.05 (s, 1H), 7.72-7.68 (m, 1H), 7.49 (d, J = 7.9 Hz, 1H), 7.31 (t, J = 7.9 Hz, 1H), 4.24 (s ,3H),3.64(s,3H),3.13(q,J=7.1Hz,2H),2.36(dd,J=7.4,4.2Hz,1H),1.36-1.29(m, 2H), 1.12 (t, J=7.2Hz, 3H), 0.88-0.79 (m, 3H), 0.74-0.68 (m, 1H). LCMS: 446.9[M+H]+.

[0150] Compound 2S: 1HNMR (400MHz, DMSO) δ11.09 (s, 1H), 10.74 (s, 1H), 8.88 (s, 1H), 8.13 (s, 1H), 8.05 (s, 1H), 7.72-7.68 (m, 1H), 7.49 (d, J = 7.9 Hz, 1H), 7.31 (t, J = 7.9 Hz, 1H), 4.24 (s ,3H),3.64(s,3H),3.13(q,J=7.1Hz,2H),2.36(dd,J=7.4,4.2Hz,1H),1.36-1.29(m, 2H), 1.12 (t, J=7.2Hz, 3H), 0.88-0.79 (m, 3H), 0.74-0.68 (m, 1H). LCMS: 446.9[M+H]+.

[0151] Example 3

[0152] Step A

[0153] Compound 1f (780 mg, 3.84 mmol) and compound 3a (500 mg, 3.85 mmol) were dissolved in a dioxane / water mixture (20 mL / 4 mL). Pd(dppf)Cl2-CH2Cl2 (157 mg, 0.19 mmol) and potassium carbonate (1.06 g, 7.68 mmol) were added sequentially. The mixture was stirred at 110°C for 16 hours under a nitrogen atmosphere. The reaction was monitored by LCMS. After completion, the mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL x 3). The combined extracts were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by medium-pressure flash chromatography (dichloromethane / methanol, 15 / 1) to afford compound 3b (300 mg, 37.44%) as an off-white solid. 1HNMR (400MHz, DMSO-d6): δppm: 8.15 (s, 1H), 7.73 (d, J = 5.2Hz, 1H), 6.96 (d, J = 5.6Hz, 1H), 6.06 (s, 2H), 3.63 (s, 3H). LCMS: 209.0 [M+H]+.

[0154] Step B

[0155] Compound 1e (65 mg, 0.23 mmol) and compound 3b (48.5 mg, 0.23 mmol) were dissolved in dioxane (5 mL). XantPhos (53.9 mg, 0.093 mmol), cesium carbonate (227.9 mg, 0.70 mmol), and Pd2(dba)3 (42.7 mg, 0.047 mmol) were added sequentially. The mixture was stirred in a microwave oven at 115°C for 2 hours under a nitrogen atmosphere. LCMS monitoring was performed. After completion of the reaction, the mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL × 3). The combined extracts were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by medium-pressure flash chromatography (dichloromethane / methanol, 20 / 1) to yield 120 mg of the crude product. The crude product was separated by preparative HPLC (acetonitrile / water = 55:45; Gemini 5uC18 150x21.2 mm) to give 30 mg of a white solid, which was then separated by SFC (Thar Preparative 80, column: CHIRALPAK IC 250 mmx20 mmx5 μm, Modifier: 40% methanol (NH4OH 0.2%) / 60% CO2, total flow rate: 40 g / min, temperature: 40°C) to give two isomers: compound 3R (11.5 mg, Rt: 3.58 min) and compound 3S (12.6 mg, Rt: 7.23 min), with a total yield of 22.93%.

[0156] Compound 3R: 1HNMR (400MHz, DMSO) δ12.32 (s, 1H), 10.72 (s, 1H), 9.68 (s, 1H), 8.95 (s, 1H), 8.32 (s, 1H), 8.17 (d, J = 5.2 Hz, 1H), 7.47 (d, J = 5.2 Hz, 1H), 3.81 (s, 3H), 3.16 (q, J = 7.2H z,2H),2.42(dd,J=7.2,4.3Hz,1H),1.44(t,J=3.6Hz,1H),1.35(dd,J=7.3,3.1Hz,1H) ,1.14(t,J=7.1Hz,3H),0.97-0.82(m,3H),0.78(d,J=5.6Hz,1H).LCMS:451.1[M+H]+.

[0157] Compound 3S: 1HNMR (400MHz, DMSO) δ12.32 (s, 1H), 10.72 (s, 1H), 9.68 (s, 1H), 8.95 (s, 1H), 8.32 (s, 1H), 8.17 (d, J = 5.2 Hz, 1H), 7.47 (d, J = 5.2 Hz, 1H), 3.81 (s, 3H), 3.16 (q, J = 7.2 Hz,2H),2.42(dd,J=7.2,4.3Hz,1H),1.44(t,J=3.6Hz,1H),1.35(dd,J=7.3,3.1Hz,1H ),1.14(t,J=7.1Hz,3H),0.97-0.82(m,3H),0.78(d,J=5.6Hz,1H).LCMS:451.2[M+H]+.

[0158] Example 4

[0159] Step A

[0160] Compound 4a (10 g, 45.66 mmol) and potassium carbonate (12.62 g, 91.33 mmol) were added sequentially to a 250 mL round-bottom flask containing 100 mL of N,N-dimethylformamide. The mixture was stirred at room temperature for 10 minutes. Deuterated iodomethane (12.96 g, 91.33 mmol) was then added and stirred at 30°C for 20 hours. The reaction was monitored by LCMS. After completion, the mixture was filtered under reduced pressure, and the filtrate was concentrated under reduced pressure. 300 mL of water was added, and the mixture was extracted with ethyl acetate (200 mL x 3). The combined extracts were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by medium-pressure flash chromatography (dichloromethane / methanol, 15 / 1) to afford compound 4b (5 g, 46.97%) as a yellow solid. LCMS: 236.0, 238.0 [M+H]+.

[0161] Step B

[0162] Compound 4b (5 g, 22.0 mmol), iron powder (4.92 g, 88.1 mmol), and ammonium chloride (4.71 g, 88.1 mmol) were added sequentially to a 500 mL round-bottom flask containing 90 mL of ethanol and 30 mL of water. The mixture was stirred at 70°C for 3 hours. After completion of the reaction, the mixture was filtered and the filtrate was concentrated under reduced pressure. The mixture was diluted with 50 mL of water and extracted with ethyl acetate (200 mL x 3). The combined extracts were dried over anhydrous Na2SO4, concentrated under reduced pressure, and separated by column chromatography (petroleum ether / ethyl acetate, 1 / 1) to afford compound 4c (3.3 g, 72.7%) as a yellow solid. 1H NMR (400 MHz, CDCl3): δ ppm: 7.60 (d, J = 5.3 Hz, 1H), 6.81 (d, J = 5.2 Hz, 1H), 4.82 (s, 2H). LCMS: 206.0, 208.0 [M+H]+.

[0163] Step C

[0164] Compound 4c (1.0 g, 4.85 mmol) and compound 1g (650 mg, 5.34 mmol) were dissolved in a mixture of dioxane (20 mL) and water (4 mL). Potassium carbonate (2.02 g, 14.55 mmol) and Pd(dppf)Cl2-CH2Cl2 (400.1 mg, 0.49 mmol) were added sequentially. The mixture was stirred at 110°C for 16 hours under a nitrogen atmosphere. LCMS monitoring was performed. After completion of the reaction, the mixture was filtered and the filtrate was concentrated under reduced pressure. The mixture was diluted with water (100 mL) and extracted with ethyl acetate (150 mL x 3). The combined extracts were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by medium-pressure flash chromatography (petroleum ether / ethyl acetate, 1 / 2) to afford compound 4d (300 mg, 29.7%) as a yellow solid.

[0165] 1HNMR (400MHz, CDCl3): δppm8.10(s,1H),7.87(d,J=5.2Hz,1H),7.18(d,J=5.2Hz,1H),4.84(s,2H),4.28(s,3H).LCMS:209.2[M+H]+.

[0166] Step D

[0167] Compound 1e (65 mg, 0.23 mmol) and compound 4d (48.5 mg, 0.23 mmol) were dissolved in dioxane (5 mL). XantPhos (53.9 mg, 0.093 mmol), cesium carbonate (227.9 mg, 0.70 mmol), and Pd2(dba)3 (42.7 mg, 0.047 mmol) were added sequentially. The mixture was stirred in a microwave oven at 115°C for 2 hours under a nitrogen atmosphere. LCMS monitoring was performed. After completion of the reaction, the mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL × 3). The combined extracts were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by medium-pressure flash chromatography (dichloromethane / methanol, 20 / 1) to yield 200 mg of the crude product. The crude product was separated by preparative HPLC (acetonitrile / water = 55:45; Gemini 5uC18 150x21.2 mm) to obtain 30 mg of product, which was then separated by SFC (Thar Preparative 80, column: CHIRALPAK IC 250mmx20mmx5μm, Modifier: 40% methanol (NH4OH 0.2%) / 60% CO2, total flow rate: 40 g / min, temperature: 40°C) to obtain two isomers: compound 4R (10.3 mg, Rt: 3.54 min) and compound 4S (9.2 mg, Rt: 7.07 min), with a total yield of 18.53%.

[0168] Compound 4R: 1HNMR (400MHz, DMSO) δ12.33 (s, 1H), 10.73 (s, 1H), 9.69 (s, 1H), 8.95 (s, 1H), 8.33 (s, 1H), 8.18 (d, J = 5.2 Hz, 1H), 7.47 (d, J = 5.2 Hz, 1H), 4.28 (s, 3H), 3.17 (q, J = 7.1H z,2H),2.42(dd,J=7.2,4.4Hz,1H),1.44(t,J=3.7Hz,1H),1.36(dd,J=7.4,3.3Hz,1H) ,1.14(t,J=7.1Hz,3H),0.95-0.82(m,3H),0.78(d,J=5.5Hz,1H).LCMS:451.1[M+H]+.

[0169] Compound 4S: 1HNMR (400MHz, DMSO) δ12.33 (s, 1H), 10.73 (s, 1H), 9.69 (s, 1H), 8.95 (s, 1H),8.33(s,1H),8.18(d,J=5.2Hz,1H),7.47(d,J=5.2Hz,1H),4.28(s,3H),3.17(q,J=7.1Hz,2H),2.42(dd,J=7.2,4.4Hz,1H),1.4 4(t,J=3.7Hz,1H),1.36(dd,J=7.4,3.3Hz,1H),1.14(t,J=7.1Hz,3H),0.95-0.82(m,3H),0.78(d,J=5.5Hz,1H).LCMS:451.1[M+H]+.

[0170] Example 5

[0171] Step A

[0172] Compound 4c (793.3 mg, 3.85 mmol) and compound 3a (500 mg, 3.85 mmol) were dissolved in a mixture of dioxane (20 mL) and water (4 mL). Potassium carbonate (1.6 g, 11.55 mmol) and Pd(dppf)Cl2-CH2Cl2 (318.5 mg, 0.39 mmol) were added sequentially. The mixture was stirred at 90°C for 16 hours under a nitrogen atmosphere. LCMS monitoring was performed. After completion of the reaction, the mixture was filtered and the filtrate was concentrated under reduced pressure. The mixture was diluted with water (100 mL) and extracted with ethyl acetate (150 mL x 3). The combined extracts were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by medium-pressure flash chromatography (petroleum ether / ethyl acetate, 1 / 2) to afford compound 5a (400 mg, 49.1%) as a yellow solid.

[0173] 1HNMR (400MHz, CDCl3): δppm 8.11 (s, 1H), 7.86 (s, 1H), 7.18 (d, J = 5.2Hz, 1H), 4.86 (s, 2H). LCMS: 212.3 [M+H]+.

[0174] Step B

[0175] Compound 1e (65 mg, 0.23 mmol) and compound 5a (48.5 mg, 0.23 mmol) were dissolved in dioxane (5 mL). XantPhos (53.9 mg, 0.093 mmol), cesium carbonate (227.9 mg, 0.70 mmol), and Pd2(dba)3 (42.7 mg, 0.047 mmol) were added sequentially. The mixture was stirred in a microwave oven at 115°C for 2 hours under a nitrogen atmosphere. LCMS monitoring was performed. After completion of the reaction, the mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL × 3). The combined extracts were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by medium-pressure flash chromatography (dichloromethane / methanol, 20 / 1) to yield 200 mg of the crude product. The crude product was separated by preparative HPLC (acetonitrile / water = 55:45; Gemini 5uC18 150x21.2 mm) to obtain 25 mg of product, which was then separated by SFC (Thar Preparative 80, column: CHIRALPAK IC 250 mmx20 mmx5 μm, Modifier: 40% methanol (NH4OH 0.2%) / 60% CO2, total flow rate: 40 g / min, temperature: 40°C) to obtain two isomers: compound 5R (4.3 mg, Rt: 3.78 min) and compound 5S (6.4 mg, Rt: 7.67 min), with a total yield of 10.14%.

[0176] Compound 5R: 1HNMR (400 MHz, DMSO) δ 12.33 (s, 1H), 10.74 (s, 1H), 9.69 (s, 1H), 8.95 (s, 1H), 8.33 (s, 1H), 8.17 (d, J = 5.2 Hz, 1H), 7.47 (d, J = 5.2 Hz, 1H), 3.17 (q, J = 7.1 Hz, 2H), 2.42 (dd, J = 7.2, 4.5 Hz, 1H), 1.44 (t, J = 3.7 Hz, 1H), 1.39-1.33 (m, 1H), 1.14 (t, J = 7.1 Hz, 3H), 0.97-0.82 (m, 3H), 0.78 (d, J = 5.4 Hz, 1H). LCMS: 454.2 [M+H]+.

[0177] Compound 5S: 1H NMR (400 MHz, DMSO) δ 12.33 (s, 1H), 10.74 (s, 1H), 9.69 (s, 1H), 8.95 (s, 1H), 8.33 (s, 1H), 8.17 (d, J = 5.2 Hz, 1H), 7.47 (d, J = 5.2 Hz, 1H), 3.17 (q, J = 7.1 Hz, 2H), 2.42 (dd, J = 7.2, 4.5 Hz, 1H), 1.44 (t, J = 3.7 Hz, 1H), 1.39-1.33 (m, 1H), 1.14 (t, J = 7.1 Hz, 3H), 0.97-0.82 (m, 3H), 0.78 (d, J = 5.4 Hz, 1H). LCMS: 454.0 [M+H]+.

[0178] Example 6

[0179] Step A

[0180] Compound 6a (300 mg, 1.59 mmol) was dissolved in concentrated sulfuric acid (4 mL) at 0°C, and fuming nitric acid (2 mL) was added dropwise. The mixture was stirred at 0°C for 1 hour. The reaction was monitored by LCMS. After completion, the reaction mixture was added to ice water (10 mL) and stirred for 5 minutes. The mixture was then extracted with ethyl acetate (50 mL x 2). The combined extracts were washed with saturated sodium bicarbonate (50 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by medium-pressure flash chromatography (dichloromethane / methanol, 20 / 1) to afford compound 6b (140 mg, 28.24%) as a yellow solid. 1H NMR (400 MHz, DMSO) δ 11.25 (s, 1H), 7.98 (s, 1H), 2.41 (s, 3H). LCMS: Rt = 1.14 min, MS 233.1 [M+H]+.

[0181] Step B

[0182] Compound 6b (140 mg, 0.60 mmol) was dissolved in N,N-dimethylformamide (5 mL), and potassium carbonate (166.0 mg, 1.20 mmol) was added. The mixture was stirred at room temperature for 0.5 hours. Under nitrogen, iodomethane (170.5 mg, 1.20 mmol) was added dropwise, and stirring was continued at room temperature for 16 hours. The reaction was monitored by LCMS. After completion, the mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL x 2). The combined extracts were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by medium-pressure flash chromatography (ethyl acetate / petroleum ether, 1 / 10) to afford compound 6c (144 mg, 97.02%) as an off-white solid. LCMS: 247.0 [M+H]+.

[0183] Step C

[0184] Compound 6c (144 mg, 0.58 mmol) was dissolved in a mixed solvent of ethanol:acetic acid:water (5 mL:5 mL:2.5 mL). Iron powder (180.8 mg, 3.23 mmol) was added at room temperature and stirred under nitrogen for 2 hours. The reaction was monitored by LCMS. After completion, the reaction was filtered under reduced pressure, and the filtrate was concentrated under reduced pressure. 30 mL of water was added, and the mixture was extracted with ethyl acetate (50 mL x 2). The combined extracts were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by medium-pressure flash chromatography (dichloromethane / methanol, 20 / 1) to afford compound 6d (120 mg, 94.81%) as a yellow solid. LCMS: MS: 217.1 [M+H]+.

[0185] Step D

[0186] Compound 6d (120 mg, 0.5527 mmol) and compound 1g (122.79 mg, 0.96 mmol) were dissolved in a mixture of dioxane (10 mL) and water (1 mL). Potassium carbonate (267.44 mg, 1.93 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride dichloromethane complex (52.67 mg, 0.06 mmol) were added sequentially. The mixture was stirred at 110°C for 16 hours under a nitrogen atmosphere. After completion of the reaction, the mixture was filtered and the filtrate was concentrated under reduced pressure. The mixture was diluted with water (100 mL) and extracted with ethyl acetate (50 mL x 2). The combined extracts were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by medium-pressure flash chromatography (dichloromethane / methanol, 20 / 1) to afford compound 6e (110 mg, 90.79%) as a yellow solid. 1HNMR (400MHz, DMSO) δ8.13(s,1H),6.83(s,1H),5.98(s,2H),4.22(s,3H),3.60(s,3H),2.24(s,3H).LCMS:MS220.2[M+H]+.

[0187] Step E

[0188] Compound 1e (33 mg, 0.12 mmol) and compound 6e (25.9 mg, 0.12 mmol) were dissolved in dioxane (5 mL). XantPhos (27.4 mg, 0.047 mmol), cesium carbonate (115.7 mg, 0.36 mmol), and Pd2(dba)3 (21.6 mg, 0.024 mmol) were added sequentially. The mixture was stirred in a microwave oven at 115°C for 2 hours under a nitrogen atmosphere. LCMS monitoring was performed. After completion of the reaction, the mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL × 3). The combined extracts were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by medium-pressure flash chromatography (dichloromethane / methanol, 20 / 1) to yield 50 mg of the crude product. The crude product was separated by preparative HPLC (acetonitrile / water, 55 / 45; Gemini 5uC18 150x21.2 mm) to obtain 20 mg of racemic product, which was then separated by SFC (Thar preparative 80, column: CHIRALPAK IC 250 mmx20 mmx5 μm, Modifier: 40% methanol (NH4OH 0.2%) / 60% CO2, total flow rate: 40 g / min, temperature: 40°C) to obtain two isomers: compound 6R (3.7 mg, Rt: 3.65 min) and compound 6S (2.5 mg, Rt: 7.82 min), with a total yield of 11.2%.

[0189] Compound 6R: 1HNMR (400MHz, DMSO) δ12.25 (s, 1H), 10.70 (s, 1H), 9.92 (s, 1H), 8.94 (s, 1H), 8.29 (s, 1H), 7.33 (s, 1H), 4.27 (s, 3H), 3.78 (s, 3H), 3.16 (q, J = 7.2 Hz, 2H), 2.51 (s, 3H), 2.42(dd,J=7.3,4.4Hz,1H),1.43(t,J=3.8Hz,1H),1.35(dd,J=7.4,3.3Hz,1H),1.13(t ,J=7.2Hz,3H),0.88(dt,J=8.4,4.8Hz,3H),0.77(d,J=5.5Hz,1H).LCMS:462.0[M+H]+.

[0190] Compound 6S: 1HNMR (400MHz, DMSO) δ12.25 (s, 1H), 10.70 (s, 1H), 9.92 (s, 1H), 8.94 (s, 1H), 8.29 (s, 1H), 7.33 (s, 1H), 4.27 (s, 3H), 3.78 (s, 3H), 3.16 (q, J = 7.2 Hz, 2H), 2.51 (s, 3H), 2.42(dd,J=7.3,4.4Hz,1H),1.43(t,J=3.8Hz,1H),1.35(dd,J=7.4,3.3Hz,1H),1.13(t ,J=7.2Hz,3H),0.88(dt,J=8.4,4.8Hz,3H),0.77(d,J=5.5Hz,1H).LCMS:462.0[M+H]+.

[0191] Example 7

[0192] Step A

[0193] Compound 1c (450 mg, 2.22 mmol) and compound 7a (370 mg, 3.33 mmol) were dissolved in dioxane (15 mL). Pd2(dba)3 (360 mg, 0.44 mmol), cesium carbonate (1.44 g, 4.41 mmol), and 1,1'-bis(dicyclohexylphosphino)ferrocene (dcpf, 335.5 mg, 0.59 mmol) were added sequentially. The mixture was stirred at 80°C for 3 hours under a nitrogen atmosphere. LCMS monitoring was performed. After completion of the reaction, the mixture was concentrated under reduced pressure, diluted with water (20 mL), and extracted with ethyl acetate (20 mL × 3). The combined extracts were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography (petroleum ether / ethyl acetate, 3 / 1) to afford compound 7b (521 mg, 84.4%) as a yellow solid. 1HNMR (400MHz, CD3OD) δ8.59(s,1H),8.33(s,1H),3.04(q,J=7.2Hz,2H),2.27(dd,J=7.4,4.2Hz,1H),1.58(t,J=3.9Hz,1 H),1.47(dd,J=7.4,3.6Hz,1H),1.19(t,J=7.2Hz,3H),1.04–0.94(m,3H),0.92–0.87(m,1H).LCMS:279.1,281.1[M+H]+.

[0194] Step B

[0195] Compound 7b (65 mg, 0.23 mmol) and compound 4d (48.54 mg, 0.23 mmol) were dissolved in dioxane (5 mL). XantPhos (54 mg, 0.093 mmol), cesium carbonate (228 mg, 0.70 mmol), and Pd2(dba)3 (42.7 mg, 0.047 mmol) were added sequentially. The mixture was stirred in a microwave oven at 115°C for 2 hours under a nitrogen atmosphere. LCMS monitoring was performed. After completion of the reaction, the mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL × 3). The combined extracts were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by medium-pressure flash chromatography (dichloromethane / methanol, 20 / 1) to yield 150 mg of the crude product. The crude product was separated by preparative HPLC (acetonitrile / water, 55 / 45; Gemini 5uC18 150x21.2 mm) to give compound 4S (30 mg, 29%) as a white solid (OR: 110, 23.2°C). 1H NMR (400 MHz, DMSO) δ 12.33 (s, 1H), 10.73 (s, 1H), 9.69 (s, 1H), 8.95 (s, 1H), 8.33 (s, 1H), 8.17 (d, J = 5.2 Hz, 1H), 7.47 (d, J = 5.2 Hz, 1H), 4.28 (s, 3H), 3.17 (q, J = 7.2 Hz, 2H) ,2.43(dd,J=7.4,4.3Hz,1H),1.44(t,J=3.8Hz,1H),1.36(dd,J=7.4,3.3Hz,1H),1.14 (t,J=7.2Hz,3H),0.98-0.83(m,3H),0.78(dd,J=8.2,2.7Hz,1H).LCMS:451.1[M+H]+.

[0196] Test Examples

[0197] Test Example 1: FACS detection of the inhibitory effect of compounds on pSTAT5 expression in CD3+ cells

[0198] 1 Experimental materials and instruments

[0199] 1.1 Experimental Reagents

[0200] DMSO, Sigma, Cat# D2650-100 mL, store at room temperature.

[0201] Perm buffer III, BD Biosciences, Cat# 558050, stored at 4°C.

[0202] Lysis / Fixation Buffer, BD Biosciences, Cat# 558049, stored at room temperature.

[0203] EDTA, Invitrogen, Cat# 15575-038, stored at room temperature.

[0204] PBS, Hydrone, Cat#SH30256.01, store at 4°C.

[0205] PE mouse anti-human CD3, BD Biosciences, Cat# 555333, store at 4°C.

[0206] Mouse anti-human phosphorylated STAT5 (pY694) (Alexa 647Conjugate), BD Biosciences, Cat#562076, stored at 4°C.

[0207] · IFN-α, Biolegend, Cat#592702.

[0208] 1.2 Experimental consumables

[0209] Microplate, 96-well, PP, v-bottom, Greiner, Cat# GN651201-100EA.

[0210] 5 mL polystyrene round-bottom tubes, FALCON, Cat# 04318011.

[0211] 96 square well storage plate, Thermo, Cat# AB-0661.

[0212] 96-well plate, Corning, Cat#3599.

[0213] 1.3 Instruments

[0214] CO2 cell incubator: MCO-15AC (Thermo).

[0215] Pipettes: 0.2-10 μL, 20-200 μL, 200-1000 μL (Thermo).

[0216] Multichannel pipettes: 0.2-10μL, 5-50μL, 20-300μL (Raining).

[0217] Centrifuge: Thermo Centrifuge ST40R; Thermo LEGEND Micro 21R.

[0218] Water system: Millipore Milli-Q reference system.

[0219] Freezer: Haier ultra-low temperature freezer.

[0220] Haier 4-degree refrigerator.

[0221] Haier -20 degrees freezer.

[0222] Eddy current: EARTH REQUIRED.

[0223] Platemaking machine: QILINBEIER; MH-2.

[0224] Flow cytometer: BD FACS flow cytometer.

[0225] 2 Experimental methods

[0226] 2.1 Compound Dilution

[0227] 1) On the day of the experiment, the compound was prepared into a 10 mM solution in DMSO, diluted to 1.5 mM in DMSO, and then diluted 3-fold in 8 steps.

[0228] concentration.

[0229] 2) Transfer 5 μL of the diluted compound to 120 μL of DPBS solution containing 0.1% BSA.

[0230] 3) Positive and negative control groups were set up, and 0.2% DMSO was added to the positive and negative control groups.

[0231] 2.2 Experimental process

[0232] 1) Add 0.5 million human PBMC cells in a volume of 67.5 uL per well of a 96-well cell culture plate.

[0233] 2) Add 3.5ul of diluted compound and mix well.

[0234] 3) Incubate in a 37°C incubator for 60 minutes.

[0235] 4) IFN-alpha was diluted to 600 ng / mL in DPBS containing 0.1% BSA, and PE-anti-hCD3 antibody was diluted to 600 ng / mL in DPBS containing 0.1% BSA.

[0236] After the 60-minute incubation, add 5 μL of diluted PE-anti-hCD3 antibody to each well.

[0237] 4uL of diluted IFN-αalpha is added to each well, meaning the final concentration of IFN-αalpha in each well is 30ng / mL.

[0238] 5) Incubate in a 37°C incubator for 30 minutes.

[0239] 6) Transfer all cells to a 96-well deep-well plate and add 1 mL of lysis / fixation buffer preheated at 37°C.

[0240] 7) Incubate at 37°C in the dark for 10 minutes.

[0241] 8) Centrifuge at 600 g for 5 minutes, discard the supernatant, add 1 mL of PBS, wash twice, and centrifuge again.

[0242] 9) Add 1 mL of Perm Buffer III to the cell pellet.

[0243] 10) Incubate at 4°C in the dark for 30 minutes.

[0244] 11) Centrifuge at 600 g for 5 minutes, discard the supernatant, add 1 mL of PBS, wash twice, and centrifuge again.

[0245] 12) Dilute APC anti-human pSTAT5 antibody 200-fold in staining buffer, add 100 μL per well to the cell wells, and mix well.

[0246] 13) Incubate at room temperature for 40 minutes.

[0247] 14) Wash twice with staining buffer (1 mL per well) and centrifuge at 600 g for 5 minutes.

[0248] 15) Discard the supernatant and resuspend the cell pellet in 300 μL staining buffer.

[0249] 16) Load the sample into a flow cytometer for analysis. Obtain the IC50 values ​​of the sample to be tested (Table 1).

[0250] Table 1

[0251] A<50nM, B>50nM.

[0252] Test Example 2: JAK family selectivity comparison

[0253] 1. Experimental Methods

[0254] 1.1. Pseudokinase assay procedures are as follows:

[0255] 1.1.1. Dissolve the compound in DMSO to a stock concentration of 10 mM.

[0256] 1.1.2. Prepare the compound dilution plate at a concentration 200 times the final concentration and dilute it 27 times.

[0257] Method, dilute from the highest concentration point to 4 concentration points in total and transfer to Echo plate.

[0258] 1.1.3. Pulse the compounds from the Echo plate into a 384-well assay plate using an Echo instrument, so that the compounds become a 3-fold serial dilution matrix with 11 concentration points.

[0259] 1.1.4. Add 5 μl of 3XTYK2 (JH2) or JAK1 (JH2) kinase to a 384-well assay plate.

[0260] 1.1.5. Add 5 μl 3XTb to a 384-well assay plate.

[0261] 1.1.6. Add 5 μl of 3XTracer to a 384-well assay plate.

[0262] 1.1.7. Centrifuge for 30 seconds and incubate at room temperature for 60 minutes.

[0263] 1.1.8. Read the signal value using Envision microplate reader (PerkinElmer).

[0264] 1.2. Kinase assay procedures are as follows:

[0265] 1.2.1. Dissolve the compound in DMSO to a stock concentration of 10 mM.

[0266] Prepare a compound dilution plate with a compound concentration 100 times the final concentration. Dilute the plate to four concentrations, starting from the highest concentration, using a 27-fold serial dilution method. Transfer the plate to an Echo plate.

[0267] 1.2.3. Pulse the compounds from the Echo plate to the 384 assay plate using the Echo instrument, so that the compounds become a 3-fold serial dilution matrix with 11 concentration points.

[0268] Prepare 2X kinase working solution and add 5 μl per well of a 384-well assay plate. Incubate the compound and kinase at room temperature for 15 minutes.

[0269] 1.2.5. Add 5ul 2X substrate (containing ATP) to a 384-well plate.

[0270] 1.2.6. Incubate at room temperature for 45 minutes.

[0271] 1.2.7. Add the detection reagent mixture to the 384-well plate, centrifuge for 30 seconds, and incubate at room temperature for 60 minutes.

[0272] 1.2.8. Read the signal using Envision microplate reader (PerkinElmer).

[0273] Data Analysis

[0274] 1.3.1. Data analysis was performed using XL-Fit software to obtain compound IC50 values ​​(Table 2).

[0275] Table 2

[0276] Test Example 3: Multiple Sclerosis Animal Model Research

[0277] Experimental Autoimmune Encephalomyelitis (EAE) is a demyelinating disease of the central nervous system and a common animal model of multiple sclerosis. This study aimed to investigate the efficacy of compound 4S on the clinical symptoms and histopathology of myelin oligodendrocyte glycoprotein (MOG)-induced EAE in mice.

[0278] Sixty specific pathogen-free (SPF) female C57BL / 6 mice were randomly divided into six groups based on body weight: a normal control group, a model (vehicle) group, a low-dose (10 mg / kg, QD) compound 4S group, a medium-dose (30 mg / kg, QD) compound 4S group, a medium-dose (30 mg / kg, BID) compound 4S group, and a high-dose (90 mg / kg, QD) compound 4S group, with 10 animals per group. Except for the normal group, mice in all other groups received a subcutaneous injection of 200 μL of MOG 35-55 emulsion to induce EAE. At 0 and 48 hours after MOG immunization, each mouse received a booster immunization with 250 μL of pertussis toxin (PTX, 1 μg / mL) intraperitoneally. Except for the normal group, mice in all other groups were administered vehicle (ethanol: vitamin E succinate polyethylene glycol ester: polyethylene glycol 300 = 5:5:90) or compound 4S daily starting 15 days after immunization (Day 15) until Day 28, for a total of 14 days. Animal health was closely monitored throughout the experiment, with body weights and clinical scores recorded daily (Figures 1A, 1B). On Day 28, mice were euthanized by carbon dioxide, and blood samples were collected and serum TNF-α and IFN-γ levels were measured by ELISA (Figures 2A, 2B).

[0279] Test Example 4: Drug Concentration Analysis of Plasma and Brain Tissue Samples

[0280] The objective of this study was to evaluate the drug concentrations in plasma and brain tissue samples of female C57BL / 6 mice following oral administration of 10 mg / kg, 30 mg / kg, and 90 mg / kg of compound 4S for 15 consecutive days. The drug concentrations of compound 4S in plasma and brain tissue homogenates were measured using liquid chromatography-mass spectrometry (LC-MS / MS) in positive ionization (ESI) multiple reaction monitoring (MRM) mode, with a linear range of 1-10,000 ng / mL.

[0281] Plasma samples from 10 mice orally administered with compound 4S: For the standard curve, quality control samples, single blank samples, and unknown samples, a 10 μL sample volume was added to each of the samples, followed by the addition of 200 μL of a 1:1 (v / v) methanol:acetonitrile precipitant (containing 5 ng / mL terfenadine). For the double blank samples, 200 μL of a 1:1 (v / v) methanol:acetonitrile precipitant was added. The samples were vortexed for 1 min and centrifuged for 15 min (4000 rpm, 4°C) to obtain the supernatant solution. The supernatant was diluted 10-fold with a 1:1 (v / v) methanol:water solution containing 0.1% FA for LC-MS / MS analysis.

[0282] Ten mouse brain tissue homogenates were orally administered with compound 4S. For the standard curve, quality control samples, single blank samples, and unknown samples, a 50 μL sample volume was added to 200 μL of a methanol:acetonitrile (1:1, v / v) precipitant (containing 5 ng / mL terfenadine). For the double blank sample, 50 μL of the blank brain tissue homogenate was added to 200 μL of a methanol / acetonitrile (1:1, v / v) precipitant. The samples were vortexed for 1 minute and centrifuged for 15 minutes (4000 rpm, 4°C) to obtain the supernatant. The supernatant was diluted 10-fold with a methanol:water (1:1, v / v, containing 0.1% FA) solution for LC-MS / MS analysis. Drug exposure in plasma and brain tissue samples was compared at different doses and at the same dose with different dosing frequencies (Table 3).

[0283] Table 3

[0284] Two hours after the last dose, a comparison of the drug concentrations in plasma and brain tissue samples at different doses revealed that the drug concentration showed a significant increase with increasing dose, demonstrating a relatively clear dose-dependence. Comparison of the same dose at different dosing frequencies revealed that the concentrations in plasma and brain tissue samples did not significantly increase with increasing dosing frequency. Furthermore, the ratio of brain tissue to plasma concentrations ranged from 0.788 to 1.32, indicating moderate brain permeability. Comparison of the dose-dependent differences in plasma concentrations revealed that the ratio between plasma concentrations was slightly lower than the dose increase, while the ratio between brain sample concentrations was slightly higher than the dose increase.

[0285] All documents mentioned in this application are incorporated herein by reference, just as if each document were incorporated herein by reference individually. It should also be understood that after reading the above teachings of the present invention, those skilled in the art may make various changes or modifications to the present invention, and that such equivalents also fall within the scope of the claims appended hereto.

Claims

1. A compound or a pharmaceutically acceptable salt thereof, characterized in that: The compound is as shown in formula A, in, Y is N or CR9; R1 is selected from the group consisting of H, optionally substituted C 1-4 alkyl; R2 is selected from the group consisting of H, optionally substituted C 1-4 alkyl; R3 and R4 are each independently selected from the group consisting of H, halogen, optionally substituted C 1-4 alkyl; R5 is selected from the group consisting of H, optionally substituted C 1-6 alkyl; R6 is selected from the group consisting of H, optionally substituted C 1-4 alkyl; R7 is selected from the group consisting of H, optionally substituted C 1-4 alkyl; R9 is selected from the group consisting of H, optionally substituted C 1-4 alkyl; The term "optionally substituted" means that the group is unsubstituted or substituted by one or more substituents selected from the group consisting of deuterium, halogen, C 1-4 Alkyl, C 1-4 Halogenated alkyl.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that Y is N or CH.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: R1 is selected from the following group: C 1-4 Alkyl, deuterated C 1-4 alkyl.

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: R2 is selected from the following group: C 1-4 Alkyl, deuterated C 1-4 alkyl.

5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: R3 is selected from the following group: H, C 1-4 alkyl.

6. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein The compound is as shown in formula I 7. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The compound is as shown in formula IS or formula IR 8. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein The compound is a compound selected from Table A or Table B or a pharmaceutically acceptable salt thereof, Table A Table B 9. The compound according to claim 8 or a pharmaceutically acceptable salt thereof, characterized in that: The compound is compound 1S, 2S, 3S, 4S, 5S, or 6S in Table B.

10. The compound according to claim 8 or a pharmaceutically acceptable salt thereof, characterized in that: The compound is compound 4S.

11. A pharmaceutical composition comprising: (i) the compound according to claim 1 or a pharmaceutically acceptable salt thereof, and (ii) a pharmaceutically acceptable carrier or excipient.

12. Use of the compound according to claim 1 or the pharmaceutical composition according to claim 5 in the preparation of (i) a medicament for treating or preventing a TYK2-mediated disease and / or (ii) a TYK2 inhibitor.

13. The use according to claim 12, characterized in that The TYK2-mediated diseases include: psoriasis, lupus erythematosus, inflammatory bowel disease, psoriatic arthritis, arthritis, vasculitis, fibrosis, dermatitis, skin aging, encephalitis, lupus nephritis, neuroinflammation, multiple sclerosis (including optic neuritis, neuromyelitis optica), chronic inflammatory demyelinating polyneuropathy, Parkinson's disease, dementia, amyotrophic lateral sclerosis, myasthenia gravis, mental illness, schizophrenia, epilepsy, spinal injury, sleep disorders, brain injury, stroke, neuropsychiatric lupus, diabetic encephalopathy, sepsis-related encephalopathy, central nervous system tumors, Huntington's disease, postoperative neurological syndrome, pain, itching, depression, narcolepsy, hydrocephalus, ankylosing spondylitis, respiratory diseases, diabetes, inflammatory eye diseases, hepatitis, cardiovascular diseases, systemic sclerosis, organ transplantation, alopecia areata, acne, eczema, vitiligo, Sjögren's syndrome, viral inflammation, cancer, or a combination thereof.

14. A method for preparing a compound, characterized in that: The compound is a compound as shown in Formula IS, And the preparation method comprises the steps of: Reacting the intermediate of formula II-S with the intermediate of formula III to obtain a compound of formula IS; In each formula, Y, R1, R2 and R3 are as defined in Formula I.

15. An intermediate of formula II-S,

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