CN112279853A - Selective JAK1 inhibitor compound and preparation method and application thereof - Google Patents

Abstract

The invention provides a heterocyclic compound serving as a JAK1 inhibitor and a synthesis and use method thereof, and particularly provides a compound shown as a formula (I), a preparation method thereof and application of the compound serving as a JAK1 inhibitor. The compounds exhibit excellent inhibitory activity against JAK 1.

Description

Selective JAK1 inhibitor compound and preparation method and application thereof

Technical Field

The invention relates to a selective JAK1 inhibitor compound, an isomer, a solvate, a salt of the compound, a medicament taking the compound or the salt as an active ingredient, and application of the compound in medicaments for treating and/or preventing JAK1 related target diseases such as immune system diseases, rheumatoid arthritis, tumors and the like.

Background

The JAK-STAT signaling pathway is a cytokine-stimulated signaling pathway discovered in recent years, JAKs play an important role in cytokine signaling, and downstream substrates of the JAK kinase family include protein-transcribing Signal Transducers and Activators (STATs). JAK proteins are important members of this pathway and abnormal increases in their activity often lead to the development of diseases, many of which are associated with abnormal cellular responses of the JAK-STAT signaling pathway, including autoimmune diseases, inflammatory diseases, bone diseases, metabolic diseases, neurological and neurodegenerative diseases, cancer, cardiovascular diseases, allergies and asthma, alzheimer's disease.

Rheumatoid Arthritis (RA) is a common chronic autoimmune disease in clinic, mainly manifested by joint swelling, pain, stiffness, deformity and severely impaired function, with a population incidence of 0.5% -1.0%. Because the pathogenesis of RA is not clear, the pathological process of RA is difficult to control, the disability rate is high, the physical and mental health of a patient is seriously damaged, and the life quality of the patient is reduced. Currently, non-steroidal anti-inflammatory drugs (NSAIDs), disease-modifying antirheumatic drugs (DMARDs), and antibody-based drugs are mainly used for the treatment of RA. First-line drugs for treating RA have long been DMARDs, and in 1988, the 1 st DMARD drug Methotrexate (MTX) was FDA approved for the treatment of RA, an important milestone in the history of RA treatment. The medicine is widely applied due to the advantages of effectiveness, tolerance, safety and the like, but has adverse reactions including nausea, vomiting, stomach discomfort, hepatotoxicity and the like. In contrast, newly developed antibody drugs have better efficacy and safety indexes for moderate-severe RA, but beneficial people are obviously limited due to targeting of specific cytokines, and the popularization of the drugs is limited due to treatment cost and injection mode administration.

Over the course of the past 20 years, RA treatment has advanced significantly and patient condition has been effectively controlled by existing treatments. Nevertheless, RA patients suffer from disease recurrence, poor therapeutic efficacy, poor long-term tolerance, and some adverse reactions. More importantly, the quality of life of RA patients, including organ function such as joints, has not been truly improved by current therapies, and there is still a great unmet clinical need in this field to address the need to restore normal function in patients.

Research shows that mononuclear/macrophage, lymphocyte and the like infiltrated in RA synovial tissues and cells generate a large amount of cytokines in an autocrine mode, the cytokines interact with each other, JAK/STAT Signal channels (Janus kinase/Signal transducer and activators of transcription signaling pathway) are activated through different ways, and cascade amplification of the cytokines can be blocked by specifically inhibiting the JAK/STAT Signal channels, so that symptoms of damaged joints of RA patients are improved, and the JAK/STAT Signal channels become potential targets for treating RA.

As JAK kinase participates in various important physiological processes in vivo, adverse reactions are possibly generated by the wide inhibition of different subtypes, Tofacitinib is used for moderate and severe RA patients with insufficient MTX reaction or intolerance, and certain adverse reactions accompanied by the Tofacitinib are observed in clinical tests and comprise infection, tuberculosis, tumor, anemia, liver injury, cholesterol increase and the like. Tofacitinib has obvious inhibitory activity on JAK1, JAK2 and JAK3 subtypes, and because JAK2 activity is related to erythrocyte differentiation and lipid metabolism, part of adverse reactions are considered to be related to the non-selective inhibitory characteristics of the medicine. Therefore, the search for selective JAK1 and/or JAK3 inhibitors would be a new direction for the study of RA drugs. At present, JAK inhibitors are proved to be used for treating diseases of a blood system, tumors, rheumatoid arthritis, psoriasis and the like.

In view of the foregoing, there remains a need in the art for the development of more selective JAK inhibitor compounds.

Disclosure of Invention

The invention aims to provide a selective JAK1 inhibitor compound with a novel structure.

In a first aspect of the present invention, there is provided a compound represented by the following formula (I), and isomers, solvates or pharmaceutically acceptable salts thereof:

wherein, the A ring is selected from the following group:

the L is selected from the following group:

R₁selected from the group consisting of: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted- (X)_m-H, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C1-3 substituents selected from N, S (O)_pAnd a heteroatom of O, a 4-10 membered heterocyclic group, a substituted or unsubstituted 6-10 membered aryl group, a substituted or unsubstituted heterocyclic group having 1-3 heteroatoms selected from N, S (O)_pAnd a 5-10 membered heteroaryl group of a heteroatom of O;

R₂、R₃、R₄and R₅Each independently selected from the group consisting of: hydrogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted- (X)_m-H, each of said X is independently selected from the group consisting of: -CH₂-，N，O，S(O)_p(ii) a And only 1,2 or 3 xs are selected from the group consisting of: n, O, S (O)_p；

Each n is independently selected from: 0.1, 2,3 or 4;

n1 is selected from: 0.1, 2,3 or 4;

n and n1 are not 0 at the same time;

n2 is selected from: 1,2 or 3;

m is selected from: 1.2, 3,4, 5 or 6;

p is 0, 1 or 2;

g is O, CH₂Or none; and when G is an O atom, n2 is selected from 2 or 3;

unless otherwise specified, "substituted" means substituted with one or more (e.g., 2,3, 4, etc.) substituents selected from the group consisting of: halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy, C3-C8 cycloalkyl, halogenated C3-C8 cycloalkyl, oxo, -CN, hydroxy, amino, carboxy, amide, sulfonamide, sulfone, a group unsubstituted or substituted with one or more substituents selected from the group consisting of: C6-C10 aryl, halogenated C6-C10 aryl, 5-10 membered heteroaryl having 1-3 heteroatoms selected from N, S and O, halogenated 5-10 membered heterocyclyl having 1-3 heteroatoms selected from N, S and O; and the substituents are selected from the group consisting of: halogen, C1-C6 alkyl, C1-C6 alkoxy, or ═ O.

In another preferred embodiment, the compound of formula I has a structure selected from the group consisting of formula I-1, I-2, I-3, and I-4:

in another preferred embodiment, the a ring is selected from the group consisting of:

in another preferred embodiment, L is selected from L1, L2, L3 and L4, preferably, L is selected from L1 and L3.

In another preferred embodiment, R is₁Selected from the group consisting of: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted- (X)_m-H, substituted or unsubstituted cycloalkyl of C3-C8, substituted or unsubstituted with 1-3 substituents selected from N, S (O)_pAnd a heteroatom of O, a 4-10 membered heterocyclic group, a substituted or unsubstituted 6-10 membered aryl group, a substituted or unsubstituted heterocyclic group having 1-3 heteroatoms selected from N, S (O)_pAnd a 5-10 membered heteroaryl group of a heteroatom of O.

In another preferred embodiment, the compound is selected from the group consisting of:

in a second aspect of the invention, there is provided a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a compound, isomer, solvate or pharmaceutically acceptable salt or hydrate thereof according to the first aspect of the invention.

In another preferred embodiment, the pharmaceutical composition is used for treating or preventing diseases related to the activity or expression amount of JAK kinase; preferably, the disease is selected from the group consisting of: cancer, myeloproliferative disease, inflammation, immunological disease, organ transplantation, viral disease, cardiovascular disease or metabolic disease, human or animal autoimmune disease, rheumatoid arthritis, skin disorders, multiple sclerosis, rheumatoid arthritis, psoriatic arthritis, inflammatory bowel disease, myasthenia gravis, psoriasis.

In a third aspect of the present invention, there is provided a use of the compound according to the second aspect of the present invention, or a pharmaceutically acceptable salt or hydrate thereof, for preparing a pharmaceutical composition for treating or preventing a disease associated with an activity or an expression amount of a JAK kinase.

In another preferred embodiment, the disease is selected from the group consisting of: cancer, myeloproliferative disease, inflammation, immunological disease, organ transplantation, viral disease, cardiovascular disease or metabolic disease, human or animal autoimmune disease, rheumatoid arthritis, skin disorders, multiple sclerosis, rheumatoid arthritis, psoriatic arthritis, inflammatory bowel disease, myasthenia gravis, psoriasis.

In another preferred embodiment, the pharmaceutical composition is for use in the treatment or prevention of a disease selected from the group consisting of: autoimmune diseases, rheumatoid arthritis, skin disorders, multiple sclerosis, rheumatoid arthritis, psoriatic arthritis, inflammatory bowel disease, myasthenia gravis, psoriasis in humans or animals.

It is to be understood that within the scope of the present invention, the above-described features of the present invention and those specifically described below (e.g., in the examples) may be combined with each other to form new or preferred embodiments. Not to be reiterated herein, but to the extent of space.

Detailed Description

The present inventors have conducted extensive and intensive studies for a long time and have unexpectedly found a compound represented by the formula (I). The compounds have unexpected activity in modulating cytokines and/or interferons and are useful in the treatment of diseases mediated by cytokines and/or interferons. Based on the above findings, the inventors have completed the present invention.

Definition of

As used herein, the term "alkyl" includes straight or branched chain alkyl groups. E.g. C₁-C₄Alkyl represents a straight or branched chain alkyl group having 1 to 4 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, and the like.

As used herein, the term "C₃-C₈Cycloalkyl "refers to cycloalkyl groups having 3 to 8 carbon atoms. It may be a single ring, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or the like. Also bicyclic, e.g., bridged, fused or spiro forms.

As used herein, the term "C₆-C₁₀Aryl "means an aryl group having 6 to 10 carbon atoms, for example, phenyl or naphthyl and the like.

As used herein, the term "5-10 membered heteroaryl having 1-3 heteroatoms selected from the group consisting of N, S and O" refers to a cyclic aromatic group having 5-10 atoms and wherein 1-3 atoms are heteroatoms selected from the group consisting of N, S and O. It may be a single ring or a condensed ring form. Specific examples may be pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, pyrrolyl, pyrazolyl, imidazolyl, (1,2,3) -triazolyl and (1,2,4) -triazolyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl and the like.

Unless specifically stated to be "substituted or unsubstituted", the groups of the present invention may be substituted with a substituent selected from the group consisting of: halogen, nitrile group, nitro group, hydroxyl group, amino group, C₁-C₆Alkyl-amino, C₁-C₆Alkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, C₁-C₆Alkoxy, halo C₁-C₆Alkyl, halo C₂-C₆Alkenyl, halo C₂-C₆Alkynyl, halo C₁-C₆Alkoxy, allyl, benzyl, C₆-C₁₂Aryl radical, C₁-C₆alkoxy-C₁-C₆Alkyl radical, C₁-C₆Alkoxy-carbonyl, phenoxycarbonyl, C₂-C₆Alkynyl-carbonyl, C₂-C₆Alkenyl-carbonyl, C₃-C₆Cycloalkyl-carbonyl, C₁-C₆Alkyl-sulfonyl, and the like.

As used herein, "halogen" or "halogen atom" refers to F, Cl, Br, and I. More preferably, the halogen or halogen atom is selected from F, Cl and Br. "halogenated" means substituted with an atom selected from F, Cl, Br, and I.

Unless otherwise specified, the structural formulae depicted herein are intended to include all isomeric forms (e.g., enantiomers, diastereomers and geometric isomers (or conformational isomers)): for example, R, S configuration containing an asymmetric center, (Z), (E) isomers of double bonds, and the like. Thus, individual stereochemical isomers of the compounds of the present invention or mixtures of enantiomers, diastereomers or geometric isomers (or conformers) thereof are within the scope of the present invention.

As used herein, the term "hydrate" refers to a complex formed by the coordination of a compound of the present invention with water.

The compounds of the present application may be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combinations of the specific embodiments with other chemical synthetic methods, and equivalents known to those skilled in the art, with preferred embodiments including, but not limited to, the examples of the present application.

The solvent used in the present application can be commercially available. Abbreviations used in this application are as follows: aq represents an aqueous solution; HATU represents O- (7-azabenzotriazol-1-yl) -N, N' -tetramethyluronium hexafluorophosphate; EDC stands for N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride; m-CPBA represents 3-chloroperoxybenzoic acid; eq represents equivalent, equivalent; CDI represents carbonyldiimidazole; DCM represents dichloromethane; PE represents petroleum ether; DIAD represents diisopropyl azodicarboxylate; DMF represents N, N-dimethylformamide; DMSO represents dimethyl sulfoxide; EtOAc for ethyl acetate; EtOH stands for ethanol; MeOH represents methanol; cbz represents benzyloxycarbonyl, an amino protecting group; boc represents tert-butyloxycarbonyl, an amino protecting group; HOAc represents acetic acid; NaCNBH₃Represents sodium cyanoborohydride; r.t. represents room temperature; THF represents tetrahydrofuran; TFA represents trifluoroacetic acid; DIPEA stands for diisopropylethylamine; boc₂O represents di-tert-butyl dicarbonate; LDA stands for lithium diisopropylamide.

The compound is artificially synthesized or

The software names, and the commercial compounds are under the supplier catalog name.

Pharmaceutical compositions and methods of administration

The compound of the present invention and various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates thereof, and pharmaceutical compositions containing the compound of the present invention as a main active ingredient are useful for the prevention and/or treatment (stabilization, alleviation or cure) of various autoimmune and inflammation-related diseases including cancer, myeloproliferative diseases, inflammation, immunological diseases, organ transplantation, viral diseases, cardiovascular diseases, metabolic diseases, and the like, because the compound of the present invention has an excellent inhibitory activity against JAK1 kinase.

The pharmaceutical compositions of the present invention comprise a safe and effective amount of a compound of the present invention in combination with a pharmaceutically acceptable excipient or carrier. Wherein "safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 1-2000mg of a compound of the invention per dose, more preferably, 1-200mg of a compound of the invention per dose. Preferably, said "dose" is a capsule or tablet.

"pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances which are suitable for human use and must be of sufficient purity and sufficiently low toxicity. By "compatible" is meant herein that the components of the composition are capable of intermixing with and with the compounds of the present invention without significantly diminishing the efficacy of the compounds. Examples of pharmaceutically acceptable carrier moieties are cellulose and its derivatives (e.g., sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (e.g., stearic acid, magnesium stearate), calcium sulfate, vegetable oils (e.g., soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (e.g., propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (e.g., propylene glycol, glycerin, mannitol, sorbitol, etc.), and the like

) Wetting agents (e.g., sodium lauryl sulfate), coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, and the like.

The mode of administration of the compounds or pharmaceutical compositions of the present invention is not particularly limited, and representative modes of administration include (but are not limited to): oral, parenteral (intravenous, intramuscular, subcutaneous or topical).

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, starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia; (c) humectants, for example, glycerol; (d) disintegrating agents, for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) slow solvents, such as paraffin; (f) absorption accelerators, e.g., quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glycerol 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 capsules, tablets and pills, the dosage forms may also comprise buffering agents.

Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared using coatings and shells such as enteric coatings and other materials well known in the art. They may contain opacifying agents and the release of the active compound or compounds in such compositions may be delayed in release in a certain part of the digestive tract. Examples of embedding components which can be used are polymeric substances and wax-like substances. If desired, the active compound may also be in microencapsulated form with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly employed in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1, 3-butylene glycol, dimethylformamide and oils, in particular, cottonseed, groundnut, corn germ, olive, castor and sesame oils or mixtures of such materials and the like.

In addition to these inert diluents, the compositions can also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

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

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 nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols and suitable mixtures thereof.

The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable therapeutic agents.

When administered in combination, the pharmaceutical composition further comprises one or more (2, 3,4, or more) other pharmaceutically acceptable therapeutic agents. One or more (2, 3,4, or more) of such other pharmaceutically acceptable therapeutic agents may be used simultaneously, separately or sequentially with a compound of the invention for the prevention and/or treatment of cytokine and/or interferon mediated diseases.

When the pharmaceutical composition is used, a safe and effective amount of the compound of the present invention is suitable for mammals (such as human beings) to be treated, wherein the administration dose is a pharmaceutically-considered effective administration dose, and for a human body with a weight of 60kg, the daily administration dose is usually 1 to 2000mg, preferably 1 to 500 mg. Of course, the particular dosage will depend upon such factors as the route of administration, the health of the patient, and the like, and is within the skill of the skilled practitioner.

The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Having described the present application in detail and having disclosed specific embodiments thereof, it will be apparent to one skilled in the art that various changes and modifications can be made to the embodiments of the application without departing from the spirit and scope of the application.

Examples

Example 1

First step of

Dissolve 1a (5.00g,32.5mmol) in acetone (50mL), cool in an ice-water bath, add 2N aqueous sodium hydroxide (19.5mL,39.0mmol) and p-toluenesulfonyl chloride (6.80g,35.7mmol) in that order, warm to room temperature after addition, and stir overnight. After the reaction was complete, filtration was carried out and the solid was washed with water (20mL × 2) and dried under reduced pressure to give 1b (8.70g) in yield: 84 percent.

MS-ESI calculated value [ M +1 ]]⁺308, measured value 308.

Second step of

1c (4.00g,23.7mmol) was dissolved in cyclohexane (200mL) and heated to 40 ℃. A solution of 2, 2-dichloroacetyl chloride (4.50g,30.8mmol) in cyclohexane (100mL) was added dropwise. After the dropwise addition, the reaction temperature was maintained at 40 ℃ and the reaction was continued for 1 hour. After completion of the reaction, the temperature was naturally lowered to room temperature, and ethyl acetate (100mL) and water (100mL) were added to the reaction system, followed by liquid separation and extraction with ethyl acetate (100 mL. times.1). The organic phases were combined, washed with saturated brine (50mL × 1), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give a residue, which was purified by column chromatography (ethyl acetate: petroleum ether ═ 0-100%) to give 1d (6.30g) in yield: 95 percent.

The third step

1d (6.30g,22.3mmol) was dissolved in methanol (100mL), and ammonium chloride (2.50g) and a Zn/Cu alloy (12.50g) were added and stirred at room temperature overnight. The solvent methanol was removed by concentration under reduced pressure, ethyl acetate (100mL) was added, filtered, and the filter cake was washed with ethyl acetate (20 mL. times.2). The organic phases were combined and concentrated under reduced pressure to give a residue which was purified by column chromatography (ethyl acetate: petroleum ether ═ 0 to 100%) to give 1e (3.00g) in yield: and 64 percent.

MS-ESI calculated value [ M-56+1 ]]⁺156, measured value 156.

The fourth step

1e (500mg,2.4mmol)) was dissolved in ethanol (10mL), and hydroxylamine hydrochloride (786mg,9.5mmol) and triethylamine (1.2g,11.9mmol) were added. Stirring overnight at room temperature, concentration under reduced pressure gave a residue which was purified by column chromatography (ethyl acetate: petroleum ether ═ 0-100%) to give 1f (450mg), yield: 84 percent.

MS-ESI calculated value [ M-56+1 ]]⁺171, found value 171.

¹H NMR(400MHz,CDCl₃)δ7.70-7.47(m,1H),4.53-4.24(m,1H),3.93-3.68(m,2H),3.54-3.33(m,1H),3.23-3.04(m,1H),2.80-2.50(m,1H),2.13-1.86(m,2H),1.46(s,9H).

The fifth step

1f (210mg,0.9mmol) was dissolved in methanol (5mL), and aqueous ammonia (0.5mL) and Raney-Ni were added. The reaction was stirred at room temperature overnight under a hydrogen atmosphere (60 psi). After completion of the reaction, the reaction mixture was directly filtered and concentrated under reduced pressure to obtain 1g (140mg) which was used as it was in the next reaction.

MS-ESI calculated value [ M +1 ]]⁺213, found 213.

The sixth step

1g of the crude product (140mg,0.9mmol) was dissolved in dichloromethane (3mL), cooled in an ice-water bath and triethylamine (202mg,2.0mmol) and p-fluorobenzoyl chloride (159mg,1.0mmol) were added. After stirring the reaction at room temperature for 1 hour, dichloromethane (50mL) and water (50mL) were added, the layers were separated, and the aqueous layer was extracted with dichloromethane (50 mL. times.1). The organic phases were combined, washed with saturated brine (50 mL. times.1), and dried over anhydrous sodium sulfate. Concentration under reduced pressure gave a residue which was purified by column chromatography (ethyl acetate: petroleum ether 0-100%) to give 1h (120mg), two-step yield: 54 percent.

MS-ESI calculated value [ M-56+1 ]]⁺279, found 279.

Seventh step

Dissolve 1h (95mg,0.3mmol) in methanol (2mL) and slowly add 4M hydrogen chloride dioxane solution (5 mL). After stirring the reaction mixture at room temperature for two hours, the reaction mixture was concentrated under reduced pressure to obtain a crude product 1i, which was used directly in the next reaction.

Eighth step

Dissolve 1i (crude from the previous step) in isopropanol (3mL) and add 1b (86mg,0.28mmol) and diisopropylethylamine (181mg, 1.4 mmol). The temperature was raised to 80 ℃ and stirred overnight. After completion of the reaction, the reaction mixture was cooled to room temperature, and concentrated under reduced pressure to give a residue, which was purified by column chromatography (ethyl acetate: petroleum ether ═ 0 to 100%) to give 1j (118mg) in two-step yield: and 69 percent.

MS-ESI calculated value [ M +1 ]]⁺506, measured value 506.

The ninth step

1j (51mg,0.1mmol) was dissolved in tetrahydrofuran (2mL), and 3N aqueous sodium hydroxide (0.14mL,0.4mmol) was added. The reaction was refluxed overnight. After completion of the reaction, the reaction mixture was cooled to room temperature, adjusted to pH 7 with 6N aqueous hydrochloric acid, and concentrated under reduced pressure to give a residue, which was purified by high performance liquid preparative chromatography to give 1(12mg) in yield: 34 percent.

MS-ESI calculated value [ M +1 ]]⁺352, found value 352.

¹H NMR(400MHz,CDCl₃)δ11.59(brs,1H),8.47(d,J＝6.0Hz,1H),8.12(s,1H),7.95(dd,J＝8.4Hz,5.6Hz,2H),7.30(t,J＝8.8Hz,2H),7.13(t,J＝2.1Hz,1H),6.53(brs,1H),4.66-4.57(m,1H),4.48-4.35(m,1H),4.08-3.79(m,2H),3.47-3.34(m,1H),2.94-2.82(m,1H),2.13-2.03(m,1H),2.03-1.89(m,2H).

Example 2

First step of

1g of the crude product (220mg,1.2mmol) was dissolved in dichloromethane (3mL), cooled in an ice-water bath and triethylamine (300mg,3.0mmol) and propanesulfonyl chloride (214mg,1.5mmol) were added. The temperature was naturally raised to room temperature, and after 1 hour of reaction, dichloromethane (50mL) and water (50mL) were added, and the mixture was separated, and the aqueous phase was extracted with dichloromethane (50 mL. times.1). The combined organic phases were washed with brine (50 mL. times.1), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (ethyl acetate: petroleum ether ═ 0 to 100%) to give 2a (135mg), yield: 42 percent.

MS-ESI calculated value [ M-56+1 ]]⁺263, found 263.

Second step of

Dissolve 2a (75mg,0.3mmol) in methanol (0.5mL) and slowly add 4M hydrogen chloride dioxane solution (2 mL). After stirring reaction for two hours at room temperature, the reaction mixture is concentrated under reduced pressure to obtain a crude product of 2b, which is directly used for the next reaction.

The third step

Dissolve 2b (crude from the previous step) in n-butanol (3mL) and add 1b (74mg,0.3mmol) and diisopropylethylamine (155mg,1.2 mmol). The temperature was raised to 130 ℃ and stirred overnight. After completion of the reaction, the reaction mixture was cooled to room temperature and concentrated under reduced pressure, and the crude product was purified by column chromatography (ethyl acetate: petroleum ether ═ 0 to 100%) to give 2c (40mg) in yield: 34 percent.

MS-ESI calculated value [ M +1 ]]⁺490, found 490.

The fourth step

2c (40mg,0.08mmol) was dissolved in tetrahydrofuran (2mL), and 3N aqueous sodium hydroxide (0.08mL,0.24mmol) was added and the reaction was refluxed overnight. After the reaction, the reaction mixture was neutralized with 6N hydrochloric acid aqueous solution, concentrated under reduced pressure, and the crude product was purified by high performance liquid preparative chromatography to give 2(8mg), yield: 29 percent.

MS-ESI calculated value [ M +1 ]]⁺336, measured value 336.

¹H NMR(400MHz,DMSO-d6)δ11.59(brs,1H),8.09(s,1H),7.44(d,J＝8.0Hz,1H),7.15-7.10(m,1H),6.50(brs,1H),4.48-4.40(m,1H),4.00-3.80(m,3H),3.30-3.20(m,1H),2.98-2.84(m,3H),2.40-2.26(m,1H),2.11-1.93(m,1H),1.86-1.74(m,1H),1.71-1.59(m,2H),0.97(t,J＝8.0Hz,3H).

Example 3

Reference is made to 1g of preparation 1a in example 1. preparation 3b from 3 a. Example 3 was prepared from 3b according to the method of example 2.

MS-ESI calculated value [ M +1 ]]⁺350, found 350.

¹H NMR(400MHz,DMSO-d6)δ11.62(s,1H),8.14(s,0.4H),8.11(s,0.6H),7.59(d,J＝8.4Hz,0.4H),7.36(d,J＝8.4Hz,0.6H),7.17-7.14(m,1H),6.68-6.67(m,0.6H),6.52-6.51(m,0.4H),4.67-4.61(m,0.6H),4.55-4.43(m,1H),4.22-4.16(m,0.4H),3.85-3.76(m,0.6H),3.69-3.61(m,0.4H),3.37-3.29(m,0.4H),3.08(t,J＝12Hz,0.6H),2.99-2.89(m,2H),2.84-2.76(m,0.6H),2.68-2.64(m,0.4H),2.59-2.52(m,0.6H),2.33-2.13(m,1.4H),1.94-1.75(m,2H),1.70-1.62(m,3.4H),1.46-1.36(m,0.6H),0.99-0.93(m,3H).

Example 4

First step of

4a (100mg,0.5mmol) was dissolved in N, N-dimethylformamide (10mL), and triethylamine (80mg,0.9mmol), cyanoacetic acid (48mg,0.6mmol) and O- (7-azabenzotriazol-1-yl) -N, N, N ', N' -tetramethylurea (233mg,0.6mmol) were added in that order. The reaction was carried out at room temperature overnight. After the reaction was completed, the reaction mixture was diluted with water (20mL), extracted with ethyl acetate (20mL × 1), and the organic phase was concentrated under reduced pressure to give a residue, which was purified by column chromatography (ethyl acetate: petroleum ether ═ 0 to 100%) to give 4b-1(88mg) in yield: 67% and 4b-2(23mg), yield: 17 percent.

Second step of

4b-1(88mg,0.3mmol) was dissolved in dichloromethane (5mL), and then trifluoroacetic acid (72mg,0.6mmol) was added to react at room temperature for 1 hour. After the reaction is finished, crude product 4c is obtained by directly concentrating under reduced pressure and is directly used for the next step.

The third step

The crude 4c (46mg,0.3mmol) was dissolved in isopropanol (5mL) and diisopropylethylamine (67mg,0.5mmol), 1a (40mg,0.3mmol) were added sequentially. The reaction mixture was reacted at 80 ℃ for 16 hours. After the reaction was completed, the temperature was lowered to room temperature, and the reaction mixture was concentrated under reduced pressure to obtain a residue, which was separated and purified by preparative high performance liquid chromatography to obtain 4(13mg) with a yield of 17%.

MS-ESI calculated value [ M +1 ]]⁺297, found 297.

¹H NMR(400MHz,CD₃OD)δ8.13(s,1H),7.13(d,J＝3.6,1H),6.73(d,J＝3.6,1H),4.39(q,1H),4.24(dd,J＝12.0,2.8Hz,1H),4.11(d,J＝11.2Hz,1H),3.89-3.60(m,2H),3.43-3.37(m,1H),3.33-3.30(m,2H),3.06-2.99(m,1H),2.74-2.66(m,1H),1.82-1.75(m,1H).

Example 5

Example 5 was obtained from compound 4b-2 via 2-step reaction according to the procedure of example 4.

MS-ESI calculated value [ M +1 ]]⁺297, found 297.

¹H NMR(400MHz,DMSO-d6)δ11.62(s,1H),8.66(d,1H),8.12(s,1H),7.13(t,J＝4.0Hz,1H),6.60-6.59(m,1H),4.13(d,J＝11.2Hz,1H),4.00(d,J＝10Hz,1H),3.99-3.93(m,1H),3.79-3.74(m,1H),3.62(s,2H),3.55-3.51(m,1H),2.98-2.91(m,2H),2.21-2.09(m,2H).

Example 6

First step of

6a (212mg,1.0mmol) was dissolved in dichloromethane (5mL) and triethylamine (202mg,2.0mmol) and p-fluorobenzoyl chloride (174mg,1.1mmol) were added sequentially. The reaction mixture was reacted at room temperature for 16 hours. Concentration under reduced pressure gave a residue, which was dispersed in ethyl acetate (10mL) and water (10mL), and the organic phase was separated and washed with saturated brine (10mL × 1), dried over anhydrous sodium sulfate solid, filtered and concentrated under reduced pressure to give a residue, which was purified by column chromatography (ethyl acetate: petroleum ether ═ 0-100%) to give 6b (250mg) in 75% yield.

MS-ESI calculated value [ M-56+1 ]]⁺279, found 279.

Second step of

6b (186mg,0.56mmol) was dissolved in dichloromethane (2.5mL) and 4M 1, 4-dioxane solution of hydrogen chloride (5mL) was added. The reaction mixture was reacted at room temperature for 2 hours, and concentrated under reduced pressure to give crude 6c (131 mg).

The third step

Crude 6c (131mg,0.56mmol) was dissolved in isopropanol (5mL) and diisopropylethylamine (289mg,2.24mmol) and 1b (206mg,0.6mmol) were added sequentially. The reaction mixture was reacted at 80 ℃ for 16 hours. After completion of the reaction, concentration under reduced pressure gave a residue, which was purified by column chromatography (ethyl acetate: petroleum ether ═ 0 to 100%) to give 6d (70mg) in 25% yield.

MS-ESI calculated value [ M +1 ]]⁺506, measured value 506.

The fourth step

6d (60mg,0.12mmol) was dissolved in tetrahydrofuran (10mL), followed by addition of 3N aqueous sodium hydroxide (20mL) and reaction at 60 ℃ for 16 hours. The organic solvent was removed by concentration under reduced pressure, the aqueous phase was adjusted to pH 7-8, extracted with ethyl acetate (10mL x 3), the resulting organic phase was washed with saturated brine (20mL x 1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a residue, which was isolated and purified by preparative liquid chromatography to give 6(16mg), yield: 37 percent.

MS-ESI calculated value [ M +1 ]]⁺352, found value 352.

¹H NMR(400MHz,CD₃OD)δ8.09(s,1H),7.86(m,2H),7.21(t,2H),7.12(d,1H),6.54(d,1H),4.51(s,2H),4.48(m,1H),4.35(s,2H),2.77(m,2H),2.43(m,2H).

Example 7

First step of

6a (212mg,1.0mmol) was dissolved in dichloromethane (5mL) and triethylamine (121mg,1.2mmol) and propanesulfonyl chloride (242mg,1.7mmol) were added. The reaction mixture was reacted at room temperature for 16 hours. After completion of the reaction, concentration under reduced pressure gave a residue, which was purified by column chromatography (ethyl acetate: petroleum ether ═ 0 to 100%) to give tert-7 a (280mg), yield: 88 percent.

Second step of

7a (260mg,0.8mmol) was dissolved in dichloromethane (2mL) and a 4M solution of hydrogen chloride in 1, 4-dioxane (10mL) was added. The reaction mixture was reacted at room temperature for 3 hours. After the reaction was complete, it was directly concentrated under reduced pressure to give crude 7b (179 mg).

The third step

Crude 7b (179mg,0.8mmol) was dissolved in n-butanol (5mL) and diisopropylethylamine (423mg,3.28mmol), 1a (151mg,1.0mmol) were added sequentially. The reaction was warmed to 120 ℃ and reacted for 16 hours. After completion of the reaction, the reaction mixture was cooled to room temperature, concentrated under reduced pressure, dissolved in water (20mL), adjusted to pH about 7 with 2N dilute hydrochloric acid, extracted with ethyl acetate (15mL × 3), the organic phases were combined, washed with saturated brine (20mL × 1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by preparative liquid chromatography to give 7(98mg) in yield: 36 percent.

MS-ESI calculated value [ M +1 ]]⁺336, measured value 336.

¹H NMR(400MHz,CD₃OD)δ8.08(s,1H),7.12(d,1H),6.51(d,1H),4.44(s,2H),4.30(s,2H),3.84(m,1H),3.02-2.98(m,2H),2.74-2.69(m,2H),2.33-2.28(m,2H),1.86-1.77(m,2H),1.08(t,3H).

Example 8

First step of

8a (350mg,1.6mmol) and triethylamine (626mg,6.2mmol) were dissolved in tetrahydrofuran (5mL), and benzyl chloroformate (528mg,3.1mmol) was added dropwise while cooling on ice. The reaction mixture was reacted at room temperature for 18 hours. After completion of the reaction, the reaction was quenched with water (10mL), extracted with dichloromethane (20mL × 2), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by column chromatography (ethyl acetate: petroleum ether ═ 0 to 100%) to obtain ester 8b (460mg) in yield: 83 percent.

MS-ESI calculated value [ M +23 ]]⁺383, found 383.

Second step of

8b (350mg,1.0mmol) was dissolved in N, N-dimethylformamide (10mL), and sodium hydride (60%, 156mg,3.9mmol) was slowly added under ice bath. The resulting reaction solution was stirred at this temperature for a further 0.5 h, then methyl iodide (552mg,3.9mmol) was added dropwise slowly. After the completion of the dropwise addition, the reaction solution was warmed to room temperature and reacted for 18 hours. After the reaction was completed, it was quenched with water (10mL), extracted with dichloromethane (20mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a residue, which was purified by column chromatography (ethyl acetate: petroleum ether ═ 0 to 100%) to give 8c (250mg), yield: 66 percent.

Calculated value of MS-ESI [ M-100+1 ]]⁺289, found value 289.

The third step

8c (250mg, 0.6mmol) was dissolved in methanol (8mL) and then wet palladium on carbon (10%, 150mg) was added. The reaction solution was placed under a hydrogen atmosphere (15psi) and stirred at room temperature overnight. After completion of the reaction, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to give a crude product 8d (150 mg).

MS-ESI calculated value [ M +1 ]]⁺255, found 255.

The fourth step

8d of the crude product (150mg, 0.6mmol), diisopropylethylamine (152mg,1.2mmol) and 1a (109mg,0.7mmol) were dissolved in isopropanol (6 mL). The reaction was stirred at 95 ℃ for 18 hours under nitrogen. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by column chromatography (ethyl acetate: petroleum ether ═ 0 to 100%) to give 8e (158mg) in yield: 72 percent.

MS-ESI calculated value [ M +1 ]]⁺372, measured value 372.

The fifth step

8e (158mg,0.4mmol) was dissolved in dichloromethane (2mL) and 4M 1, 4-dioxane hydrogen chloride solution (6mL) was added. The reaction solution was stirred at room temperature for 3 hours. After the reaction was completed, the reaction mixture was directly concentrated under reduced pressure to obtain crude product 8f (115mg), yield: 100 percent.

MS-ESI calculated value [ M +1 ]]⁺272, found value 272.

The sixth step

The 8f crude product (115mg,0.4mmol), triethylamine (128mg,1.3mmol) was dissolved in dichloromethane (3 mL). Propane-1-sulfonyl chloride (73mg,0.5mmol) was added dropwise at 0 deg.C under nitrogen. The reaction mixture was reacted at room temperature for 4 hours. After completion of the reaction, the reaction was quenched with saturated brine (5mL), extracted with dichloromethane (10mL × 2), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give a residue, which was purified by preparative high performance liquid chromatography to give 8(36mg), yield: 23 percent.

MS-ESI calculated value [ M +1 ]]⁺378, measured value 378.

¹H-NMR(400MHz,CDCl₃)δ10.03(s,1H),8.30(s,1H),7.04(d,J＝3.6Hz,1H),6.55(d,J＝3.2Hz,1H),5.25-5.15(m,1H),4.29-4.23(m,1H),3.36(s,3H),2.87-2.83(m,5H),2.52-2.44(m,2H),2.34-2.29(m,5H),2.20-2.17(m,1H),1.83-1.71(m,2H),1.08-1.04(m,3H).

Example 9

First step of

9a (56mg,0.2mmol) was dissolved in N, N-dimethylformamide (2mL), sodium hydride (60%, 9mg,0.2mmol) was added at 0 ℃ under nitrogen, stirring was continued for 10 min after the addition was complete, and methyl iodide (34mg,0.3mmol) was added. The reaction solution was warmed to room temperature and the reaction was continued for 1 hour. After the reaction was complete, the reaction was quenched with saturated aqueous ammonium chloride (10mL), extracted with ethyl acetate (20mL x 2), and the organic phases were combined, washed with saturated brine (20mL x 1), dried over anhydrous sodium sulfate, filtered, and concentrated to give crude 9b (50 mg). Directly used for the next reaction.

MS-ESI calculated value [ M-56+1 ]]⁺242, found value 242.

Second step of

9b (742mg,2.5mmol) was dissolved in dichloromethane (10mL) and trifluoroacetic acid (10mL) was added. The reaction was stirred at room temperature overnight. After the reaction was completed, crude 9c (500mg) was directly obtained by concentration under reduced pressure and used in the next reaction.

MS-ESI calculated value [ M-56+1 ]]⁺198, found 198.

The third step

Crude 9c (500mg,2.5mmol) was dissolved in isopropanol (10mL) and diisopropylethylamine (970mg,7.5mmol) and 1b (750mg,2.5mmol) were added. The reaction solution was heated to 90 ℃ and reacted for 48 hours. After the reaction was completed, it was cooled to room temperature, and concentrated under reduced pressure to give a residue, which was purified by column chromatography (ethyl acetate: petroleum ether ═ 0 to 100%) to give 9d (300mg), yield in 3 steps: 28 percent.

MS-ESI calculated value [ M-56+1 ]]⁺469, found 469.

¹H NMR(400MHz,CDCl₃)δ8.38(s,1H),8.05-8.03(d,J＝8Hz,2H),7.45-7.44(d,J＝4Hz,1H),7.28-7.26(d,J＝8Hz,2H),6.61-6.60(d,J＝4.4Hz,1H),5.00-4.97(m,1H),4.15(q,J＝7.2Hz,14.4Hz,2H),3.19(s,3H),3.06-3.01(m,1H),2.44-2.38(m,3H),2.34(s,3H),2.32-2.29(m,2H),2.22-2.15(m,3H),1.27(t,J＝7.6Hz,14.4Hz,3H).

The fourth step

9d (1.2g,1.8mmol) was dissolved in ethanol/water (20mL/20mL ═ v/v), and sodium hydroxide (222mg,5.4mmol) was added. The reaction solution was heated to 90 ℃ and reacted for 1 hour. After completion of the reaction, the reaction mixture was cooled to room temperature, adjusted to pH 3 with 1N hydrochloric acid, extracted with dichloromethane (50mL × 3), and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give 9e (500mg) which was used in the next reaction.

The fifth step

The crude 9e (20mg,0.07mmol) was dissolved in dichloromethane (10m L) and 1-ethyl- (3-dimethylaminopropyl) carbonyldiimine hydrochloride (27mg,0.14mmol), 1-hydroxybenzotriazole (19mg,0.14mmol) and 2,2, 2-trifluoroethylamine (10mg,0.1mmol) were added under nitrogen. The reaction mixture was stirred at room temperature for 1 hour. After the reaction was complete, dichloromethane (10mL) and water (10mL) were added, the layers were separated, the aqueous phase was extracted with dichloromethane (10mL x 1), the organic phases were combined, washed with saturated brine (20mL x 1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a residue which was preparative purified by reverse phase to give 9(5mg), yield: 18 percent.

MS-ESI calculated value [ M +1 ]]⁺368, measured value 368.

¹H NMR(400MHz,DMSO-d6)δ11.61(s,1H),8.37(t,J＝6.4Hz,12.4Hz,1H),8.09(s,1H),7.14(t,J＝2.8Hz,5.6Hz,1H),6.54(d,J＝1.6Hz,1H),5.13-5.09(m,1H),3.89-3.84(m,2H),3.19(s,3H),3.02-2.98(m,1H),2.36-2.33(m,1H),2.29-2.25(m,3H),2.19-2.15(m,3H),2.12-2.09(m,1H).

Example 10

Example 10 was synthesized from 9e according to the procedure of example 9.

MS-ESI calculated value [ M +1 ]]⁺380, found 380.

¹H NMR(400MHz,DMSO-d6)δ11.61(s,1H),9.98(s,1H),8.10(s,1H),7.63-7.60(d,J＝12Hz,1H),7.30-7.29(m,2H),7.14-7.13(m,1H),6.86-6.81(m,1H),6.56-6.55(m,1H),5.16-5.10(m,1H),3.21(s,3H),3.19-3.11(m,1H),2.43-2.13(m,8H).

Example 11

Example 11 was synthesized from 9e according to the procedure of example 9.

MS-ESI calculated value [ M +1 ]]⁺363, measured value 363.

¹H NMR(400MHz,CD₃OD-d4)δ8.17-8.16(m,1H),8.16-7.98(m,2H),7.68-7.63(m,1H),7.00-6.97(m,2H),6.54-6.53(d,J＝3.6Hz,1H),5.03-4.95(m,1H),3.17(s,3H),3.16-3.15(m,1H),2.48-2.13(m,8H).

Example 12

The crude 9e (100mg,0.4mmol) was dissolved in N, N-dimethylformamide (5mL) and N, N-diisopropylethylamine (90mg,0.7mmol), cyclopropylamine (40mg,0.7mmol) and 2- (7-benzotriazol oxide) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (135mg,0.35mmol) were added at room temperature. The reaction solution was stirred at room temperature for 12 hours. After completion of the reaction, ethyl acetate (10mL) and water (10mL) were added, and the mixture was separated and extracted with ethyl acetate (10 mL. times.1). The organic phases were combined, washed with brine (20mL x 1), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue which was purified by reverse phase preparative purification to give 12(13mg) yield: 11 percent.

MS-ESI calculated value [ M +1 ]]⁺326, found 326.

¹H NMR(400MHz,DMSO-d6)δ11.60(s,1H),8.09(s,1H),7.72(d,J＝4.4Hz,1H),7.13-7.12(m,1H),6.54-6.53(m,1H),5.13-5.08(m,1H),3.19(s,3H),2.87-2.61(m,1H),2.60-2.56(m,1H),2.36-2.12(m,7H),2.12-1.98(m,1H),0.61-0.51(m,2H),0.35-0.32(m,2H).

Example 13

Example 13 was synthesized from 9e according to the procedure of example 11.

MS-ESI calculated value [ M +1 ]]⁺376, found 376.

¹H NMR(400MHz,DMSO-d6)δ11.60-11.55(brs,1H),8.16-8.10(m,2H),7.12(s,1H),6.55(s,1H),5.14-5.08(m,1H),4.05-3.99(m,1H),3.18(s,3H),2.90-2.83(m,3H),2.65-2.60(m,1H),2.38-2.01(m,9H).

Example 14

Example 14 was synthesized from 9e according to the procedure of example 9.

MS-ESI calculated value [ M +1 ]]⁺380, found 380.

¹H NMR(400MHz,DMSO-d₆)δ11.61(s,1H),9.82(s,1H),8.10(s,1H),7.63-7.60(m,2H),7.14-7.10(m,3H),6.56-6.55(m,1H),5.19-5.10(m,1H),3.22(s,3H),3.18-3.10(m,1H),2.45-2.12(m,8H).

Example 15

First step of

15a (50mg,0.2mmol) and triethylamine (67mg,0.7mmol) were dissolved in dichloromethane (10mL) and 4-fluorobenzoyl chloride (53mg,0.3mmol) was added at 0 ℃ under nitrogen. The reaction solution was stirred at room temperature for 3 hours. After completion of the reaction, the reaction mixture was quenched with saturated brine (20mL), extracted with dichloromethane (20mL × 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give a residue, which was purified by column chromatography (ethyl acetate: petroleum ether ═ 0-100%) to give 15b (70mg) in yield: 91 percent.

MS-ESI calculated value [ M +1 ]]⁺349, found 349.

Second step of

15b (70mg,0.2mmol) was dissolved in dichloromethane (10mL) and trifluoroacetic acid (2mL) was added dropwise at 0 ℃. The reaction solution was stirred at room temperature for 1 hour. After the reaction was completed, the reaction mixture was directly concentrated under reduced pressure to give crude 15c (50mg), yield: 100 percent.

The third step

The crude product 15c (50mg,0.2mmol), diisopropylethylamine (106mg,0.8mmol) and 1b (63mg,0.2mmol) were dissolved in isopropanol (10 mL). The reaction was stirred at 90 ℃ overnight under nitrogen. After the reaction was completed, it was cooled to room temperature, concentrated under reduced pressure, and the residue was diluted with a saturated sodium bicarbonate solution (20mL), extracted with ethyl acetate (30mL × 3), combined with the organic phases, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a residue, which was purified by column chromatography (methanol: dichloromethane ═ 0 to 100%) to give 15d (100mg) in yield: 93 percent.

MS-ESI calculated value [ M +1 ]]⁺520, found 520.

The fourth step

15d (100mg,0.2mmol) and sodium hydroxide (39mg,1.0mmol) were dissolved in a mixed solvent of tetrahydrofuran and water (20mL/20mL ═ v/v). The reaction was stirred at 75 ℃ overnight under nitrogen. After the reaction, the reaction mixture was cooled to room temperature, and the pH of the reaction mixture was adjusted to about 7 with 1N hydrochloric acid. The precipitated solid was filtered, and the residue was washed with tetrahydrofuran (10mL × 3), water (10mL × 3), and acetonitrile (10mL × 3) in this order, and the residue was dried under reduced pressure to give 15(40mg), yield: 57 percent.

MS-ESI calculated value [ M +1 ]]⁺366, found value 366.

¹H-NMR(400MHz,DMSO-d6)δ11.51(s,1H),8.42(d,J＝8.0Hz,1H),8.22(s,1H),7.91-7.83(m,2H),7.25(t,J＝8.0Hz,2H),7.10(t,J＝4.0Hz,1H),6.60(q,J＝1.6Hz,1H),4.47-4.36(m,1H),3.91-3.86(m,2H),3.82-3.76(m,2H),2.77(s,2H),2.36-2.23(m,2H),1.59-1.51(m,2H).

Example 16

First step of

14a (50mg,0.2mmol) and triethylamine (67mg,0.7mmol) were dissolved in dichloromethane (10mL) and propane-1-sulfonyl chloride (63mg,0.4mmol) was added dropwise at 0 deg.C under nitrogen. The reaction was stirred at room temperature overnight. After the reaction was completed, the reaction was quenched with saturated brine (20mL), extracted with dichloromethane (20mL × 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give a residue, which was purified by column chromatography (ethyl acetate: petroleum ether ═ 0-100%) to give 16a (46mg) in yield: 62 percent.

MS-ESI calculated value [ M-56+1 ]]⁺277, found 277.

Second step of

16a (46mg, 0.14mmol) was dissolved in dichloromethane (10mL) and trifluoroacetic acid (2mL) was added dropwise at 0 ℃. The resulting reaction solution was stirred at room temperature for 1 hour. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain crude 16b (32mg) in yield: 100 percent.

The third step

Crude product to 16b (32mg,0.14mmol), diisopropylethylamine (71mg,0.6mmol) and 1b (43mg,0.14mmol) were dissolved in isopropanol (10 mL). The reaction was stirred at 90 ℃ overnight under nitrogen. After the reaction was completed, it was cooled to room temperature, and concentrated under reduced pressure to give a residue, which was diluted with a saturated sodium bicarbonate solution (20mL), extracted with ethyl acetate (30mL × 3), combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a residue, which was purified by column chromatography (methanol: dichloromethane ═ 0 to 100%) to give 16c (60mg), yield: 86 percent.

MS-ESI calculated value [ M +1 ]]⁺504, found 504.

The fourth step

16c (60mg,0.12mmol) and sodium hydroxide (24mg,0.6mmol) were dissolved in a mixed solvent of tetrahydrofuran and water (20mL/10mL ═ v/v). The reaction was stirred at 75 ℃ overnight under nitrogen. After completion of the reaction, the reaction mixture was cooled to room temperature, 1N hydrochloric acid was added dropwise to adjust the pH to 7, and the precipitated solid was filtered and washed with tetrahydrofuran (10 mL. times.3), water (10 mL. times.3), and acetonitrile (10 mL. times.3) in this order. The residue was dried under reduced pressure to give 16(40mg), yield: 95 percent.

MS-ESI calculated value [ M +1 ]]⁺350, found 350.

¹H-NMR(400MHz,DMSO-d6)δ11.57(s,1H),8.08(s,1H),7.19(d,J＝8.0Hz,1H),7.11(t,J＝2.8Hz,1H),6.58(d,J＝2.4Hz,1H),3.87-3.82(m,2H),3.73-3.68(m,3H),2.95(t,J＝8.0Hz,2H),2.68(t,J＝3.2Hz,2H),2.29-2.21(m,2H),1.65(t,J＝7.6Hz,2H),1.45-1.37(m,2H),0.97(t,J＝7.6Hz,3H).

Example 17

First step of

17a (4mL,0.11mmol) was mixed with acetic anhydride (2.5mL,0.03mmol) under nitrogen and stirred at 60 deg.C for 1 h. The reaction was cooled to afford 17b crude product (6.5mL), yield: 100 percent.

Second step of

17c (50mg,0.2mmol) was dissolved in dichloromethane (5mL) and 17b (1mL) was added dropwise thereto at 0 ℃. The reaction solution was stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was quenched with 1N sodium hydroxide solution (5mL), extracted with dichloromethane (10mL × 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure and filtered to obtain 17d of a crude product (50mg), yield: 89 percent.

MS-ESI calculated value [ M-56+1 ]]⁺199, found 199.

The third step

17d of the crude product (50mg,0.2mmol) was dissolved in tetrahydrofuran (10mL) and borane dimethylsulfide (1M,0.6mL,0.6mmol) was added dropwise thereto at 0 ℃ under nitrogen. The reaction was stirred at room temperature overnight. After the reaction was completed, it was quenched with methanol (2mL), diluted with 1N sodium hydroxide solution (5mL), extracted with dichloromethane (10mL × 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 17e crude product (45mg), yield: 88 percent.

MS-ESI calculated value [ M +1 ]]⁺241, measured value 241.

The fourth step

The 17e crude product (45mg,0.2mmol), diisopropylethylamine (72mg,0.6mmol) and 1a (28mg,0.2mmol) were dissolved in n-butanol (10 mL). The reaction was stirred at 120 ℃ overnight under nitrogen. After completion of the reaction, it was cooled to room temperature, and concentrated under reduced pressure to give a residue, which was purified by column chromatography (dichloromethane: petroleum ether ═ 0 to 100%) to give 17f (55mg) in yield: 82 percent.

MS-ESI calculated value [ M +1 ]]⁺358, found 358.

The fifth step

17f (55mg,0.15mmol) was dissolved in anhydrous dichloromethane (10mL), to which trifluoroacetic acid (1mL) was added dropwise at 0 ℃. The reaction solution was stirred at room temperature for 1 hour. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain 17g of a crude product (39mg), yield: 100 percent.

MS-ESI calculated value [ M +1 ]]⁺258, found 258.

The sixth step

17g of the crude product (39mg,0.15mmol) and triethylamine (31mg,0.3mmol) were dissolved in dichloromethane (10 mL). 4-fluorobenzoyl chloride (24mg,0.15mmol) was added dropwise thereto at 0 deg.C under nitrogen. The reaction solution was stirred at room temperature for 2 hours. After the reaction was completed, the reaction was quenched with saturated brine (20mL), extracted with dichloromethane (10mL × 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give a residue, which was purified by preparative high performance liquid chromatography to give 17(15mg) in yield: 26 percent.

MS-ESI calculated value [ M +1 ]]⁺380, found 380.

¹H-NMR(400MHz,DMSO-d6)δ11.61(s,1H),8.61(d,J＝7.6Hz,1H),8.10(s,1H),7.92(dd,J＝8.8Hz,2.0Hz,2H),7.29(t,J＝8.8Hz,2H),7.14(t,J＝2.8Hz,1H),6.56(d,J＝0.9Hz,1H),5.21-5.16(m,1H),4.39-4.32(m,1H),3.22(s,3H),2.43-2.12(m,8H).

Example 18

First step of

17c (50mg,0.2mmol) and triethylamine (67mg,0.7mmol) were dissolved in dichloromethane (10mL) and propane-1-sulfonyl chloride (63mg,0.4mmol) was added dropwise thereto at 0 ℃ under a nitrogen blanket. The reaction was stirred at room temperature overnight. After the reaction was completed, the reaction was quenched with saturated brine (20mL), extracted with dichloromethane (20mL × 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give a residue, which was purified by column chromatography (ethyl acetate: petroleum ether ═ 0-100%) to give 18a (71mg) in yield: 97 percent.

MS-ESI calculated value [ M +23 ]]⁺355, found 355.

Second step of

18a (71mg,0.21mmol) was dissolved in anhydrous tetrahydrofuran (5mL) and lithium aluminum hydride (24mg, 0.63mmol) was added portionwise thereto at 0 ℃ under nitrogen. The reaction was stirred at 80 ℃ overnight. After the reaction was complete, cool to room temperature, quench the reaction carefully with water (1mL), stir vigorously, filter, and concentrate the filtrate under reduced pressure to give 18b crude product (50mg), yield: 96 percent.

MS-ESI calculated value [ M +1 ]]⁺247 measured value247。

The third step

18b crude (50mg,0.2mmol), diisopropylethylamine (105mg,0.8mmol) and 1a (31mg,0.2mmol) were dissolved in n-butanol (10 mL). The reaction was stirred at 120 ℃ overnight under nitrogen. After the reaction was completed, it was cooled to room temperature, and concentrated under reduced pressure to give a residue, which was subjected to preparative high performance liquid chromatography to give 18(60mg) in yield: 14 percent.

MS-ESI calculated value [ M +1 ]]⁺364, measured value 364.

¹H-NMR(400MHz,DMSO-d6)δ11.60(s,1H),8.09(s,1H),7.37(d,J＝8.8Hz,1H),7.13(t,J＝2.8Hz,1H),6.55(d,J＝1.6Hz,1H),5.16-5.11(m,1H),3.68-3.61(m,1H),3.20(s,3H),2.91-2.87(m,2H),2.38-2.31(m,5H),2.29-2.23(m,1H),2.17-1.97(m,2H),1.69-1.62(m,2H),0.98(t,J＝7.6Hz,3H).

Example 19

First step of

4a (110mg,0.52mmol) was dissolved in dichloromethane (5mL), and then triethylamine (158mg,1.56mmol) and p-fluorobenzoyl chloride (164mg,1.04mmol) were sequentially added to the reaction system, reacted at room temperature for 1 hour, and the reaction solution was concentrated under reduced pressure to give a residue, which was purified by column chromatography (ethyl acetate: petroleum ether ═ 0-100%) to give 19a (160mg) in yield: 92 percent.

Second step of

To a solution of 19a (160mg,0.48mmol) in dichloromethane (5mL) was added a 4M solution of dioxane hydrochloride (0.50mL,2.00mmol), reacted at room temperature for 1 hour, and concentrated under reduced pressure directly to give crude 19b (112mg) which was used in the next reaction.

The third step

The crude product of 19b (112mg,0.48mmol) was dissolved in isopropanol (5mL), and diisopropylethylamine (185mg,1.44mmol), 1a (73mg,0.48mmol) were added to the reaction system in this order, and the temperature was raised to 80 ℃ for reaction for 16 hours. After naturally cooling to room temperature, the reaction system was diluted with water (20mL) and extracted with ethyl acetate (10 mL. times.3), the organic phases were combined and concentrated under reduced pressure to give a residue, which was purified by preparative HPLC to give 19-Peak1(62mg), two-step yield: 37% and 19-Peak2(20mg), two-step yield: 12 percent.

19-Peak1：

MS-ESI calculated value [ M +1 ]]⁺352, found value 352.

¹H-NMR(400MHz,DMSO-d6)δ11.57(s,1H),8.57(d,J＝6.0Hz,1H),8.08(s,1H),7.90(dd,J＝8.4Hz,5.6Hz,2H),7.27(t,J＝8.8Hz,2H),7.08(t,J＝3.0Hz,1H),6.52(s,1H),4.46-4.38(m,1H),4.12(m,1H),4.00(m,1H),3.60-3.51(m,2H),3.33-3.30(m,1H),2.92(m,1H),2.58-2.53(m,1H),2.06-1.99(m,1H).

19-Peak2：

MS-ESI calculated value [ M +1 ]]⁺352, found value 352.

¹H-NMR(400MHz,DMSO-d6)δ11.63(s,1H),8.80(d,J＝6.8Hz,1H),8.13(s,1H),7.95(dd,J＝8.8Hz,5.6Hz,2H),7.31(t,J＝8.8Hz,2H),7.15(s,1H),6.62(s,1H),4.23-4.19(m,2H),4.07-4.03(m,1H),3.84-3.78(m,1H),3.59-3.55(m,1H),3.12-3.07(m,1H),3.03-2.99(m,1H),2.43-2.33(m,1H),2.18-2.14(m,1H).

Example 20

First step of

Dissolving 6a (212mg,1.00mmol) in N, N-dimethylformamide (5mL), then adding triethylamine (303mg,3.00mmol), 2- (7-benzotriazole oxide) -N, N' -tetramethylurea hexafluorophosphate (570mg,1.50mmol), cyanoacetic acid (128mg,1.50mmol) in this order, stirring the reaction system at room temperature for 16 hours, adding water (20mL) to quench the reaction, extracting the reaction solution with ethyl acetate (20mL × 3), combining the organic phases, concentrating under reduced pressure to obtain a residue, purifying the residue by column chromatography (ethyl acetate: petroleum ether ═ 0-100%) to obtain 20a (140mg), yield: 50 percent.

Second step of

To a solution of 20a (120mg,0.43mmol) in dichloromethane (5mL) was added trifluoroacetic acid (1mL) slowly dropwise, reacted at room temperature for 3 hours, and concentrated under reduced pressure to give crude 20b (90mg) which was used directly in the next reaction.

The third step

20b (40mg,0.22mmol) was dissolved in n-butanol (2mL), then triethylamine (68mg,0.67mmol), 1a (34mg,0.22mmol) were added in this order, the temperature was raised to 100 ℃ for microwave reaction for 1.5 hours, the reaction was concentrated under reduced pressure to give a residue, which was purified by preparative HPLC to give 20(21mg), yield: 32 percent.

MS-ESI calculated value [ M +1 ]]⁺297, found 297.

¹H-NMR(400MHz,DMSO-d₆)δ11.60(s,1H),8.51(d,J＝7.2Hz,1H),8.08(s,1H),7.13(s,1H),6.37(s,1H),4.30(s,2H),4.19(s,2H),4.14-4.08(m,1H),3.59(s,2H),2.59-2.54(m,2H),2.19-2.13(m,2H).

Example 21

First step of

17g (29mg,0.11mmol), 3,3, 3-trifluoropropionic acid (29mg,0.22mmol) and triethylamine (22mg,0.22mmol) were dissolved in N, N-dimethylformamide (4mL), and O- (7-azabenzotriazol-1-yl) -N, N, N ', N' -tetramethylurea (84mg,0.22mmol) was added and reacted at room temperature overnight. The reaction was quenched by addition of water (20mL) and extracted with ethyl acetate (10mL x 2), the organic phases were combined, washed with saturated brine (10mL x 1), the organic phase was concentrated under reduced pressure to give a residue which was purified by preparative HPLC to give 21(21mg), yield: 51 percent.

MS-ESI calculated value [ M +1 ]]⁺368, measured value 368.

¹H-NMR(400MHz,CD₃OD)δ8.11(s,1H),7.12(d,J＝4.0Hz,1H),6.65(d,J＝3.6Hz,1H),5.16-5.07(m,1H),4.30-4.22(m,1H),3.33-3.31(m,3H),3.17-3.09(m,2H),2.66-2.60(m,1H),2.53-2.48(m,1H),2.44-2.29(m,4H),2.14-2.02(m,2H).

Example 22

First step of

To a solution of 17g (22mg,0.08mmol), triethylamine (16mg,0.16mmol) in dichloromethane (5mL) was added dropwise propionyl chloride (17mg,0.16mmol), reacted at room temperature for 1 hour, and concentrated under reduced pressure to give a residue which was purified by preparative HPLC to give 22(6mg), yield: 23 percent.

MS-ESI calculated value [ M +1 ]]⁺326, found 326.

¹H-NMR(400MHz,CDCl₃)δ9.95(s,1H),8.28(s,1H),7.04(d,J＝4.0Hz,1H),6.56(d,J＝3.2Hz,1H),5.73-5.71(m,1H),5.21-5.16(m,1H),4.92(s,1H),4.43-4.33(m,1H),3.31(m,3H),2.68-2.62(m,1H),2.50-2.40(m,2H),2.34-2.28(m,3H),2.04-1.93(m,2H),0.97-0.95(m,2H),0.74-0.72(m,2H).

Example 23

First step of

To a solution of 17g (29mg,0.11mmol), triethylamine (22mg,0.22mmol) in dichloromethane (5mL) was added dropwise 3-fluorobenzoyl chloride (36mg,0.22mmol), reacted at room temperature for 1 hour, and concentrated under reduced pressure to give a residue, which was purified by preparative HPLC to give 23(11mg), yield: 27 percent.

MS-ESI calculated value [ M +1 ]]⁺380, found 380.

¹H-NMR(400MHz,DMSO-d₆)δ11.61(s,1H),8.68(d,J＝7.6Hz,1H),8.10(s,1H),7.70(d,J＝7.6Hz,1H),7.65(d,J＝10.4Hz,1H),7.51(q,J＝5.6Hz,1H),7.40-7.35(m,1H),7.14(d,J＝3.6Hz,1H),6.57(d,J＝3.2Hz,1H),5.23-5.14(m,1H),4.39-4.32(m,1H),3.22(s,3H),2.43-2.37(m,2H),2.35-2.28(m,3H),2.24-2.13(m,3H).

Example 24

First step of

Dissolving 9e (80mg,0.28mmol), 24a (28mg,0.34mmol) and 1-hydroxybenzotriazole (76mg,0.56mmol) and sodium bicarbonate (47mg,0.56mmol) in dichloromethane (5mL), adding 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (108mg,0.56mmol) to the reaction system, reacting overnight at room temperature, adding water (20mL) to the reaction solution to quench the reaction, extracting with dichloromethane (20mL × 2), combining the organic phases, washing with saturated saline (10mL × 1), concentrating the organic phase under reduced pressure to obtain a residue, and purifying the residue by preparative HPLC to obtain 24(6mg), yield: 6 percent.

MS-ESI calculated value [ M +1 ]]⁺353, measured value 353.

¹H-NMR(400MHz,DMSO-d6)δ11.61(s,1H),10.97(s,1H),8.10(s,1H),7.84(s,1H),7.13(d,J＝3.6Hz,1H),7.07(s,1H),6.55(d,J＝3.6Hz,1H),5.17-5.08(m,1H),3.29-3.28(m,1H),3.21(s,3H),2.41-2.29(m,4H),2.27-2.14(m,4H).

Example 25

First step of

Example 25(18mg) was obtained by reacting 9e with morphine according to the procedure of example 9, yield: 21 percent.

MS-ESI calculated value [ M +1 ]]⁺356, found 356.

¹H-NMR(400MHz,DMSO-d6)δ11.61(s,1H),8.09(s,1H),7.14-7.13(m,1H),6.55-6.54(m,1H),5.12-5.08(m,1H),3.54-3.51(m,4H),3.43-3.42(m,2H),3.34-3.33(m,2H),3.29-3.21(m,1H),3.20(s,3H),2.43-2.36(m,1H),2.34-2.09(m,7H).

Example 26

Example 3 purification by chiral SFC gave examples 26-Peak1, 26-Peak2, 26-Peak3, 26-Peak 4.

26-Peak1：

MS-ESI calculated value [ M +1 ]]⁺350, found 350.

¹H NMR(400MHz,DMSO-d6)δ11.61(s,1H),8.10(s,1H),7.362(d,J＝8.0Hz,1H),7.15(d,J＝3.6Hz,1H),6.67(d,J＝2.4Hz,1H),4.67-4.61(m,1H),4.47(d,J＝12.4Hz,1H),3.81(t,J＝7.2Hz,1H),3.07(t,J＝12.4Hz,1H),2.97-2.92(m,2H),2.82-2.77(m,1H),2.58-2.49(m,1H),2.16(t,J＝20.2,10.0Hz,1H),1.94-1.62(m,5H),1.43-0.99(m,1H),0.96(t,J＝7.2Hz,1H).

26-Peak2：

MS-ESI calculated value [ M +1 ]]⁺350, found 350.

¹H NMR(400MHz,DMSO-d6)δ11.63(s,1H),8.14(s,1H),7.59(s,1H),7.17(d,J＝3.6Hz,1H),6.52(d,J＝3.6Hz,1H),4.55-4.50(m,1H),4.22-4.16(m,1H),3.68-3.62(m,1H),3.37-3.36(m,1H),2.91(t,J＝15.6Hz,2H),2.67-2.50(m,1H),2.33-2.22(m,2H),1.86-1.84(m,1H),1.71-1.62(m,5H),0.95(t,J＝7.2Hz,3H).

26-Peak3：

MS-ESI calculated value [ M +1 ]]⁺350, found 350.

¹H NMR(400MHz,DMSO-d6)δ11.62(s,1H),8.14(s,1H),7.59(d,J＝7.6Hz，1H),7.17(d,J＝3.6Hz,1H),6.52(d,J＝3.6Hz,1H),4.55-4.50(m,1H),4.22-4.16(m,1H),3.68-3.62(m,1H),3.37-3.36(m,1H),2.91(t,J＝15.6Hz,2H),2.67-2.50(m,1H),2.33-2.22(m,2H),1.86-1.84(m,1H),1.71-1.62(m,5H),0.95(t,J＝7.2Hz,3H).

26-Peak4：

MS-ESI calculated value [ M +1 ]]⁺350, found 350.

¹H NMR(400MHz,DMSO-d6)δ11.61(s,1H),8.10(s,1H),7.36(d,J＝8.0Hz,1H),7.15(d,J＝3.6Hz,1H),6.67(d,J＝2.4Hz,1H),4.67-4.61(m,1H),4.47(d,J＝12.4Hz,1H),3.81(t,J＝7.2Hz,1H),3.07(t,J＝12.4Hz,1H),2.97-2.92(m,2H),2.82-2.77(m,1H),2.58-2.49(m,1H),2.16(t,J＝20.2,10.0Hz,1H),1.94-1.62(m,5H),1.43-0.99(m,1H),0.96(t,J＝7.2Hz,1H).

Example 27

First step of

To a solution of 17g (7mg,0.03mmol), oxazole-4-carboxylic acid (4mg,0.03mmol) and triethylamine (6mg,0.06mmol) in N, N-dimethylformamide (3mL) was added O- (7-azabenzotriazol-1-yl) -N, N, N ', N' -tetramethylurea (21mg,0.06mmol), and the mixture was reacted at room temperature overnight. The reaction was quenched by addition of water (20mL), extracted with ethyl acetate (10mL x 2), the organic phase was washed with saturated brine (10mL x 1), the organic phase was concentrated under reduced pressure to give a residue, which was purified by preparative HPLC to give 27(4mg), yield: 40 percent.

MS-ESI calculated value [ M +1 ]]⁺353, measured value 353.

¹H-NMR(400MHz,CDCl₃)δ9.89(s,1H),8.31(s,1H),8.23(s,1H),7.86(s,1H),7.04(d,J＝3.6Hz,2H),6.56(d,J＝3.6Hz,1H),5.26-5.17(m,1H),4.58-4.48(m,1H),3.30(s,3H),2.74-2.69(m,1H),2.56-2.46(m,2H),2.38-2.29(m,3H),2.17-2.07(m,2H).

Example 28

First step of

Example 28(5mg) was obtained by reacting 9e with 2-aminothiazole in accordance with the method of example 9, yield: 8 percent.

MS-ESI calculated value [ M +1 ]]⁺369, found 369.

¹H-NMR(400MHz,CD₃OD)δ8.14(s,1H),7.44-7.43(s,1H),7.12-7.11(s,2H),6.66(s,1H),5.19-5.10(m,1H),3.35(m,4H),2.53-2.35(m,8H).

Example 29

First step of

9e (100mg,0.35mmol) was dissolved in pyridine (5mL), 2, 3-difluoroaniline (91mg,0.70mmol) was added, the temperature was reduced to 0 ℃, phosphorus oxychloride (60mg,0.39mmol) was added to the reaction system, and after stirring at room temperature for 12 hours, concentration under reduced pressure gave a residue which was purified by preparative HPLC to give 29(19mg), yield: 14 percent.

MS-ESI calculated value [ M +1 ]]⁺398, found 398.

¹H-NMR(400MHz,DMSO-d6)δ11.61(s,1H),9.75(s,1H),8.10(s,1H),7.68-7.66(m,1H),7.18-7.13(m,3H),6.56-6.55(m,1H),5.16-5.12(m,1H),3.28(s,1H),3.21(s,3H),2.42-2.14(m,8H).

Example 30

Example 30(2mg) was obtained by reacting 9e and 2-amino-1, 3, 4-thiadiazole according to the procedure of example 9, yield: 2 percent.

MS-ESI calculated value [ M +1 ]]⁺370, found 370.

¹H-NMR(400MHz,CDCl₃ and CD₃OD)δ8.76(s,1H),8.13(s,1H),6.97(s,1H),6.49(s,1H),5.10(m,1H),3.33-3.25(m,4H),2.54-2.19(m,8H).

Example 31

Example 31(11mg) was obtained by reacting 9e with 2,2,3,3, 3-pentafluoropropylamine according to the procedure of example 9, yield: 8 percent.

MS-ESI calculated value [ M +1 ]]⁺418, actual measurementA value 418.

¹H-NMR(400MHz,DMSO-d6)δ11.68(s,1H),8.35-8.32(m,1H),8.09(s,1H),7.13-7.12(m,1H),6.55-6.53(m,1H),5.13-5.09(m,1H),3.98-3.88(m,2H),3.19(s,3H),3.04-2.99(m,1H),2.48-2.13(m,8H).

Example 32

First step of

Example 32(15mg) was obtained by reacting 9e with 1,1,1,3,3, 3-hexafluoro-2-propylamine according to the method of example 29, yield: 10 percent.

MS-ESI calculated value [ M +1 ]]⁺436, found 436.

¹H-NMR(400MHz,DMSO-d6)δ11.60(s,1H),9.19-9.16(d,J＝10.0Hz,1H),8.10(s,1H),7.14-7.12(m,1H),6.55-6.54(m,1H),5.89-5.83(m,1H),5.13-5.09(m,1H),3.20(s,3H),3.18-3.14(m,1H),2.39-2.16(m,8H).

Example 33

Example 33(23mg) was obtained by reacting 9e with 4-trifluoromethylthiazol-2-amine according to the method of example 29, in yield: 15 percent.

MS-ESI calculated value [ M +1 ]]⁺437, found 437.

¹H-NMR(400MHz,DMSO-d6)δ12.46(s,1H),11.61(s,1H),8.10(s,1H),7.93(s,1H),7.14-7.13(m,1H),6.56-6.55(m,1H),5.15-5.11(m,1H),3.28-3.21(m,1H),3.19(s,3H),2.42-2.19(m,8H).

Example 34

Example 34(3mg) was obtained by reacting 9e with 3-methyl-1, 2, 4-thiadiazol-5-amine according to the method of example 29, yield: 5 percent.

MS-ESI calculated value [ M +1 ]]⁺384, found 384.

¹H-NMR(400MHz,DMSO-d6)δ12.75-12.64(brs,1H),11.60(s,1H),8.10(s,1H),7.14-7.11(m,1H),6.55-6.53(m,1H),5.17-5.09(m,1H),3.32-3.26(m,1H),3.20(s,3H),2.43(s,3H),2.42-2.21(m,8H).

Example 35

Example 35(6mg) was obtained by reacting 9e with 3-methoxy-1, 2, 4-thiadiazol-5-amine according to the method of example 29, yield: 9 percent.

MS-ESI calculated value [ M +1 ]]⁺400, found value 400.

¹H-NMR(400MHz,DMSO-d6)δ12.79-12.70(brs,1H),11.60(s,1H),8.10(s,1H),7.14-7.12(m,1H),6.56-6.55(m,1H),5.17-5.08(m,1H),3.92(s,3H),3.37-3.33(m,1H),3.20(s,3H),2.42-2.21(m,8H).

Example 36

Example 36(14mg) was obtained by reacting 9e with 3, 3-difluoropyrrolidine according to the procedure of example 9, yield: 25 percent.

MS-ESI calculated value [ M +1 ]]⁺376, found 376.

¹H-NMR(400MHz,CDCl₃)δ9.68(s,1H),8.22(s,1H),6.96-6.95(m,1H),6.48-6.48(m,1H),5.12-5.04(m,1H),3.76-3.70(m,1H),3.66-3.60(m,2H),3.56-3.52(m,1H),3.21(s,3H),3.08-2.94(m,1H),2.48-2.13(m,10H).

Example 37

Example 37(18mg) was obtained by reacting 9e with 4-cyanopiperidine according to the procedure of example 9 in yield: 32 percent.

MS-ESI calculated value [ M +1 ]]⁺379, found 379.

¹H-NMR(400MHz,CDCl₃)δ9.68(s,1H),8.22(s,1H),6.96-6.95(m,1H),6.48-6.48(m,1H),5.12-5.04(m,1H),3.83-3.78(m,1H),3.77-3.61(m,2H),3.37-3.29(m,4H),3.23-3.14(m,1H),2.91-2.86(m,1H),2.54-2.16(m,8H),1.92-1.73(m,4H).

Example 38

Example 38(4mg) was obtained by reacting 9e with 2-aminoacetonitrile according to the procedure of example 9, yield: 8 percent.

MS-ESI calculated value [ M +1 ]]⁺325, found value 325.

¹H-NMR(400MHz,CDCl₃)δ9.68(s,1H),8.22(s,1H),6.96-6.95(m,1H),6.48-6.48(m,1H),5.75-5.28(s,1H),5.18-5.10(m,1H),4.24-4.15(m,2H),3.28(s,3H),3.00-2.92(s,1H),2.54-5.10(m,8H).

Example 39

Example 39(4mg) was obtained by reacting 9e with methylpiperazine according to the procedure of example 9, yield: 7 percent.

MS-ESI calculated value [ M +1 ]]⁺369, found 369.

¹H-NMR(400MHz,CDCl₃)δ9.68(s,1H),8.22(s,1H),6.96-6.95(m,1H),6.48-6.48(m,1H),5.19-5.10(m,1H),3.67-3.44(m,2H),3.38-3.33(m,1H),3.32(s,3H),3.30-3.14(m,1H),2.54-2.12(m,16H).

Example 40

Example 40(2mg) was obtained by reacting 9e with N-methyl-2, 2, 2-trifluoroethylamine according to the procedure of example 9, yield: 4 percent.

MS-ESI calculated value [ M +1 ]]⁺382, measured value 382.

¹H-NMR(400MHz,CDCl₃)δ9.04(s,1H),8.21(s,1H),7.30-7.26(m,1H),7.24-6.55(m,1H),5.20-5.13(m,1H),4.07-4.00(m,2H),3.33-3.21(m,4H),3.05-3.02(s,3H),2.56-2.11(m,8H).

EXAMPLE 41

Example 41(9mg) was obtained by reacting 9e with ethylamine according to the method of example 9, yield: 20 percent.

MS-ESI calculated value [ M +1 ]]⁺314, measured value 314.

¹H-NMR(400MHz,CDCl₃)δ9.14-9.13(m,1H),8.28(s,1H),7.01-7.00(m,1H),6.55-6.54(m,1H),5.30(s,1H),5.16-5.11(m,1H),3.33-3.26(m,5H),2.90-2.86(m,1H),2.52-2.11(m,8H),1.25-1.14(m,3H).

Example 42

Example 42(2mg) was obtained by reacting 9e with 3-methanesulfonylazetidine according to the procedure of example 9, yield: 3 percent.

MS-ESI calculated value [ M +1 ]]⁺404, measured value 404.

¹H-NMR(400MHz,CDCl₃ and CD₃OD)δ8.16(s,1H),7.04-7.03(m,1H),6.55(s,1H),5.07-5.06(m,1H),4.44-4.40(m,2H),4.29-4.27(m,2H),3.71(s,1H),3.28-3.27(m,3H),3.04-2.94(m,4H),2.44-2.18(m,8H).

Example 43

Example 43(9mg) was obtained by reacting 9e with 4, 4-difluoropiperidine according to the procedure of example 9, yield: 15 percent.

MS-ESI calculated value [ M +1 ]]⁺390, found 390.

¹H-NMR(400MHz,CDCl₃)δ9.72(s,1H),8.29(s,1H),7.03-7.02(m,1H),6.56-5.65(m,1H),5.18-5.14(m,1H),3.73-3.68(m,2H),3.48-3.45(m,2H),3.28(s,3H),3.23-3.19(m,1H),2.54-2.15(m,8H),2.15-1.91(m,4H).

Example 44

Example 44(1mg) was obtained by reacting 9e with thiomorpholine 1, 1-dioxide according to the procedure of example 9, yield: 2 percent.

MS-ESI calculated value [ M +1 ]]⁺404, measured value 404.

¹H-NMR(400MHz,DMSO-d6)δ11.61(s,1H),8.09(s,1H),7.14-7.12(m,1H),6.55-6.54(m,1H),6.56-5.6.55(m,1H),5.11(m,1H),3.86-3.72(m,4H),3.35-3.33(m,3H),3.29-3.08(m,4H),2.42-2.11(m,8H).

Example 45

First step of

Dissolving 9b (500mg,1.70mmol) in tetrahydrofuran (5mL), cooling to-78 ℃, adding 1M lithium diisopropylamide tetrahydrofuran solution (0.25mL,2.50mmol) dropwise to the reaction system, stirring at-78 ℃ for 1 hour, adding methyl iodide (350mg,2.50mmol) to the system, continuing stirring for 2 hours, concentrating under reduced pressure to give a residue, and purifying the residue by column chromatography (petroleum ether: ethyl acetate ═ 0-100%) to give 45a (320mg) with yield: and 63 percent.

Example 45 was obtained from 45a via a multi-step reaction according to the procedure for synthesizing example 9 from 9b in example 9.

MS-ESI calculated value [ M +1 ]]⁺382, measured value 382.

¹H-NMR(400MHz,DMSO-d₆)δ11.60(s,1H),8.18(t,J＝6.4Hz,1H),8.09(s,1H),7.14-7.12(m,1H),6.55-6.52(m,1H),5.13-5.08(m,1H),3.93-3.80(m,2H),3.19(s,3H),2.43-2.25(m,4H),2.20-2.14(m,2H),2.05-2.00(m,1H),1.93-1.87(m,1H),1.30(s,3H).

Example 46

Example 46(18mg) was obtained by reacting 9e with 5-methoxythiazol-2-amine according to the method of example 9, yield: 27 percent.

MS-ESI calculated value [ M +1 ]]⁺399, measured value 399.

¹H-NMR(400MHz,DMSO-d6)δ11.61(s,1H),8.10(s,1H),7.13-7.12(d,J＝3.6Hz,1H),6.76(s,1H),6.55-6.54(d,J＝3.2Hz,1H),5.14-5.10(m,1H),3.81(s,3H),3.20(s,3H),3.18-3.14(m,1H),2.39-2.14(m,8H).

Example 47

Example 47(7mg) was obtained by reacting 9e and 5- (trifluoromethyl) thiazol-2-amine according to the method of example 9, yield: 10 percent.

MS-ESI calculated value [ M +1 ]]⁺437, found 437.

¹H-NMR(400MHz,DMSO-d6)δ12.61(s,1H),11.61(s,1H),8.10-8.08(m,2H),7.14-7.13(m,1H),6.56-6.55(m,1H),5.15-5.11(m,1H),3.34(s,1H),3.21(s,3H),2.41-2.22(m,8H).

Example 48

Example 48(5mg) was obtained by reacting 9e with 4- (tert-butyl) thiazol-2-amine according to the method of example 9, yield: 7 percent.

MS-ESI calculated value [ M +1 ]]⁺425, found 425.

¹H-NMR(400MHz,DMSO-d6)δ11.94(s,1H),11.61(s,1H),8.10(s,1H),7.14-7.12(m,1H),6.71(s,1H),6.56-6.55(m,1H),5.15-5.11(m,1H),3.29-3.22(m,1H),3.20(s,3H),2.41-2.15(m,8H),1.25(s,9H).

Example 49

Example 49(8mg) was obtained by reacting 9e with 5-methylthiazol-2-amine according to the method of example 9, yield: 12 percent.

MS-ESI calculated value [ M +1 ]]⁺383, found 383.

¹H-NMR(400MHz,DMSO-d6)δ11.76(brs,1H),11.61(s,1H),8.10(s,1H),7.14-7.10(m,2H),6.56-6.54(m,1H),5.17-5.09(m,1H),3.29-3.23(m,1H),3.20(s,3H),2.41-2.15(m,8H),1.24(s,3H).

Example 50

Example 50(2mg) was obtained by reacting 9e and 5- (trifluoromethyl) isoxazol-3-amine according to the method of example 29, yield: 4 percent.

MS-ESI calculated value [ M +1 ]]⁺421, measured value 421.

¹H-NMR(400MHz,DMSO-d6)δ11.62(s,1H),11.29(s,1H),8.10(s,1H),7.62(s,1H),7.14-7.13(m,1H),6.56(s,1H),5.14-5.10(m,1H),3.26-3.21(m,1H),3.20(s,3H),2.41-2.17(m,8H).

Example 51

Example 51(9mg) was obtained by reacting 9e with (3R,4S) -3, 4-difluoropyrrolidine hydrochloride according to the procedure of example 9 in yield: 14 percent.

MS-ESI calculated value [ M +1 ]]⁺376, found 376.

¹H-NMR(400MHz,DMSO-d6)δ11.67(s,1H),8.09(s,1H),7.14-7.12(m,1H),6.55-6.54(m,1H),5.38-5.33(m,1H),5.24-5.22(m,1H),5.21-5.08(m,1H),3.82-3.41(m,4H),3.19(s,3H),3.18-3.16(m,1H),2.42-2.31(m,8H).

Example 52

Example 51(5mg) was obtained by reacting 9e with (3R,4R) -3, 4-difluoropyrrolidine according to the procedure of example 9, yield: 9 percent.

MS-ESI calculated value [ M +1 ]]⁺376, found 376.

¹H-NMR(400MHz,CDCl₃)δ8.28(s,1H),7.05-7.04(d,J＝4.0Hz,1H),6.58-6.57(d,J＝4.0Hz,1H),5.22-5.09(m,3H),3.99-3.89(m,1H),3.80-3.66(m,3H),3.31(s,3H),3.15-3.10(m,1H),2.55-2.18(m,9H).

Example 53

Example 53(4mg) was obtained by reacting 17g with 4- (trifluoromethyl) thiazole-2-carboxylic acid according to the method of example 17, yield: 5 percent.

MS-ESI calculated value [ M +1 ]]⁺437, found 437.

¹H-NMR(400MHz,DMSO-d6)δ11.60(s,1H),9.28(d,J＝8.4Hz,1H),8.76(d,J＝0.8Hz,1H),8.10(s,1H),7.14-7.12(m,1H),6.56-6.55(m,1H),5.18-5.14(m,1H),4.38-4.32(m,1H),3.21(s,3H),2.43-2.12(m,8H).

Example 54

Reference example 3 example 54-Peak1 and 54-Peak2 were obtained from 3b and 54a by a multi-step reaction.

54-Peak1：

MS-ESI calculated value [ M +1 ]]⁺409, measured value 409.

¹H NMR(400MHz,DMSO-d6)δ11.63(s,1H),9.20(d,J＝6.8Hz,1H),8.14(s,1H),7.86(t,J＝9.6Hz,1H),7.61-7.57(m,1H),7.17(t,J＝3.2Hz,1H),6.59(t,J＝1.6Hz,1H),4.84-4.78(m,1H),4.22-4.15(m,1H),3.99-3.94(m,1H),3.71-3.65(m,1H),2.73(t,J＝6.0Hz,1H),2.51-2.34(m,2H),1.94-1.63(m,4H).

54-Peak2：

MS-ESI calculated value [ M +1 ]]⁺409, measured value 409.

¹H NMR(400MHz,DMSO-d6)δ11.63(s,1H),8.88(d,J＝6.8Hz,1H),8.12(s,1H),7.87-7.83(m,1H),7.57-7.53(m,1H),7.16(t,J＝3.2Hz,1H),6.69(t,J＝1.6Hz,1H),4.85-4.78(m,1H),4.45(d,J＝12.4Hz,1H),4.37-4.29(m,1H),3.20-3.14(m,1H),2.94(t,J＝7.2Hz,1H),2.61-2.57(m,1H),2.33-2.24(m,1H),1.94(d,J＝3.6Hz,1H),1.79-1.68(m,2H),1.51-1.43(m,4H).

Biological activity assay

The in vitro inhibition effect of the compound on JAK1 kinase is detected by using a Caliper mobility shift assay method. Test compounds were dissolved in DMSO to prepare 10mM stock solutions. Gradient dilution of compound stock solution with DMSO to prepare 50X working solution (total 10 concentrations), and transferring 40 μ L of each working solution to

In the motherboard. By using

A non-contact nano-liter acoustic pipetting system transfers 400nL of compound solution or DMSO at a corresponding concentration from a master plate to a 384-well reaction plate. And then 10 muL of 2% kinase solution is added into a 384-hole reaction plate, after the reaction plate is incubated for 10 minutes at room temperature, 10 muL of 2% FAM labeled polypeptide and ATP mixed solution is added, after the reaction plate is incubated for a specific time at 28 ℃,30 muL of stop solution is added, detection is carried out on a Caliper, and the conversion value is calculated, namely the height of the product peak is higher than the sum of the heights of the substrate peak and the product peak. Percent inhibition of kinase by the compound was calculated using the following formula and an IC was fitted using XLFIT5.4.0.8₅₀The value is obtained.

Percent inhibition is (max-conversion)/(max-min) × 100, where "max" represents the conversion reading for the DMSO control and "min" represents the conversion reading for the low control. The results of the testing of the synthesized compounds of the present invention are shown in table 1.

Inhibitory Activity of the Compounds of Table 1 against JAK1

Note: 0.01 μ M +++ < ═ 0.01 μ M +++ <0.1 μ M, and +++ > <0.1 μ M. Indicates inhibitory activity of the indicated compounds against JAK1 at 1mM ATP concentration.

And (4) conclusion: the compound synthesized by the invention has strong inhibiting effect on Jak 1.

All documents referred to herein are incorporated by reference into this application as if each were individually incorporated by reference. Furthermore, it should be understood that various changes and modifications of the present invention can be made by those skilled in the art after reading the above teachings of the present invention, and these equivalents also fall within the scope of the present invention as defined by the appended claims.

Claims (10)

1. A compound of formula (I), and isomers, solvates or pharmaceutically acceptable salts thereof:

wherein, the A ring is selected from the following group:

the L is selected from the following group:

R₁selected from the group consisting of: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted- (X)_m-H, substituted or unsubstituted cycloalkyl of C3-C8, substituted or unsubstituted with 1-3 substituents selected from N, S (O)_pAnd a heteroatom of O, a 4-10 membered heterocyclic group, a substituted or unsubstituted 6-10 membered aryl group, a substituted or unsubstituted heterocyclic group having 1-3 heteroatoms selected from N, S (O)_pAnd a 5-10 membered heteroaryl group of a heteroatom of O;

R₂、R₃、R₄and R₅Each independently selected from the group consisting of: hydrogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted- (X)_m-H, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, each said X being independently selected from the group consisting of: -CH₂-，N，O，S(O)_p(ii) a And only 1,2 or 3 xs are selected from the group consisting of: n, O, S (O)_p；

Each n is independently selected from: 0.1, 2,3 or 4;

n1 is selected from: 0.1, 2,3 or 4;

n and n1 are not 0 at the same time;

n2 is selected from: 1.2 or 3;

m is selected from: 1.2, 3,4, 5 or 6;

p is 0, 1 or 2;

g is O, CH₂Or none; and when G is an O atom, n2 is selected from 2 or 3;

unless otherwise specified, "substituted" means substituted with one or more (e.g., 2,3, 4, etc.) substituents selected from the group consisting of: halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy, C3-C8 cycloalkyl, halogenated C3-C8 cycloalkyl, oxo, -CN, hydroxy, amino, carboxy, amide, sulfonamide, sulfone, a group unsubstituted or substituted with one or more substituents selected from the group consisting of: C6-C10 aryl, halogenated C6-C10 aryl, 5-10 membered heteroaryl having 1-3 heteroatoms selected from N, S and O, halogenated 5-10 membered heterocyclyl having 1-3 heteroatoms selected from N, S and O; and the substituents are selected from the group consisting of: halogen, C1-C6 alkyl, C1-C6 alkoxy, or ═ O.

2. The compound of claim 1, wherein the compound of formula I has a structure selected from the group consisting of formula I-1, I-2, I-3, and I-4:

3. the compound of claim 1, wherein the a ring is selected from the group consisting of:

4. the compound of claim 1, wherein L is selected from the group consisting of L1, L2, L3 and L4, preferably L is selected from the group consisting of L1 and L3.

5. The compound of claim 1, wherein the compound is characterized byIn the above formula R₁Selected from the group consisting of: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted- (X)_m-H, substituted or unsubstituted cycloalkyl of C3-C8, substituted or unsubstituted with 1-3 substituents selected from N, S (O)_pAnd a heteroatom of O, a 4-10 membered heterocyclic group, a substituted or unsubstituted 6-10 membered aryl group, a substituted or unsubstituted heterocyclic group having 1-3 heteroatoms selected from N, S (O)_pAnd a 5-10 membered heteroaryl group of a heteroatom of O.

6. The compound of claim 1, wherein said compound is selected from the group consisting of:

7. a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a compound, isomer, solvate or pharmaceutically acceptable salt or hydrate thereof of claim 1.

8. The pharmaceutical composition of claim 7, wherein the pharmaceutical composition is for treating or preventing a disease associated with the activity or expression level of a JAK kinase; preferably, the disease is selected from the group consisting of: cancer, myeloproliferative disease, inflammation, immunological disease, organ transplantation, viral disease, cardiovascular disease or metabolic disease, human or animal autoimmune disease, rheumatoid arthritis, skin disorders, multiple sclerosis, rheumatoid arthritis, psoriatic arthritis, inflammatory bowel disease, myasthenia gravis, psoriasis.

9. Use of a compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt or hydrate thereof, for the preparation of a pharmaceutical composition for the treatment or prevention of a disease associated with the activity or expression level of a JAK kinase.

10. The use according to claim 9, wherein the disease is selected from the group consisting of: cancer, myeloproliferative disease, inflammation, immunological disease, organ transplantation, viral disease, cardiovascular disease or metabolic disease, human or animal autoimmune disease, rheumatoid arthritis, skin disorders, multiple sclerosis, rheumatoid arthritis, psoriatic arthritis, inflammatory bowel disease, myasthenia gravis, psoriasis.