Phosphodiesterase 4B inhibitor as well as preparation method and application thereof

Abstract

The invention provides a phosphodiesterase 4B selective inhibitor as well as a preparation method and application thereof. Specifically, the invention provides a compound as shown in a formula (I), or a stereoisomer, a tautomer, a solvate, a prodrug, an isotope label and pharmaceutically acceptable salts thereof, and a pharmaceutical composition containing the compound. The invention also provides application of the compound and / or the pharmaceutical composition in preparation of drugs for preventing and / or treating inflammatory diseases and fibrosis diseases. The compound and / or the pharmaceutical composition provided by the invention show relatively high phosphodiesterase 4B inhibitory activity. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the field of medicinal chemistry, and in particular to a class of selective phosphodiesterase 4B inhibitors and a preparation method and use thereof. Background Art

[0002] Inflammation is a very common and important basic pathological process. The defensive response of living tissues with vascular systems to damaging factors is called inflammation. Inflammatory reactions are closely related to many diseases, such as dermatitis, psoriasis, systemic lupus erythematosus, etc. For example, systemic lupus erythematosus is caused by genetic changes, which causes epidermal hyperplasia, thereby causing a series of inflammatory reactions. Therefore, treating inflammation plays an important role in treating the above diseases. Studies have shown that various inflammatory reactions in the body are related to the molar concentration of intracellular cAMP. cAMP and cGMP play an important regulatory role in cell activity, and the regulation of their concentration is mainly determined by the balance between the synthesis of nucleotide cyclase and the hydrolysis of phosphodiesterase (PDEs).

[0003] Phosphodiesterases (PDEs) have the function of hydrolyzing the intracellular second messengers cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP). A decrease in the intracellular cAMP or cGMP content will cause abnormal biochemical reactions transmitted by these second messengers. PDEs are widely distributed in the human body, and their physiological effects involve multiple research fields. Excessive levels of PDEs in the body will cause a decrease in cAMP concentration, thereby inducing diseases such as asthma, depression, psoriasis, and inflammation. PDEs have 11 subtypes, one of which, PDE4, plays a significant role in inflammatory cells. In recent years, PDEs, as a new therapeutic target, have attracted widespread attention from many scholars and become a new research hotspot.

[0004] There are 4 subtypes of PDE4, namely PDE4A to PDE4D. Existing PDE4 inhibitors such as rolipram, roflumilast, cilomilast, apremilast, etc. are all non-selective PDE4 inhibitors. Due to poor selectivity, existing PDE4 inhibitors cause a variety of drug side effects, especially the inhibition of PDE4D in the central nervous system, which can cause severe vomiting reactions. International patent application WO2013026797 discloses a new type of PDE4B inhibitor-Compound II, but further studies have found that Compound II is not highly selective for PDE4D, and vomiting side effects have occurred in clinical trials. Therefore, it is of great clinical significance to find and discover PDE4B inhibitors with better selectivity.

[0005] Summary of the invention

[0006] In order to overcome the deficiencies of the prior art, the present invention provides a compound having the following formula (I):

[0007]

[0008] or its stereoisomers, tautomers, solvates, prodrugs, isotope-labeled substances, and pharmaceutically acceptable salts thereof, wherein

[0009] X is selected from S=O, S(=O)2;

[0010] R1 is selected from H, C1-C6 alkyl;

[0011] R1' is selected from C3-C8 carbocyclic group, C6-C 10 aryl, 5-10 membered heteroaryl, 5-10 membered heterocyclic group; wherein the carbocyclic group, aryl, heteroaryl, heterocyclic group is replaced by R4-(CR a R b ) m -substituted and optionally substituted by halogen, hydroxy, amino, mercapto, C1-C6 alkyl, C1-C6 alkoxy;

[0012] R2, R2', R3, R3' are each independently selected from hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl;

[0013] R4 is selected from hydroxyl, amino, thiol, carboxyl, -SO3H, -PO4H, tetrazolyl, triazolyl, -C(=O)OC1-C6 alkyl, -C(=O)NR c R d ;

[0014] R a , R b are each independently selected from hydrogen, C1-C6 alkyl, halogen, hydroxyl, or R a , R b Together with the carbon atom to which it is attached, it forms a C3-C6 carbocyclic group; R c , R d are each independently selected from hydrogen, C1-C6 alkyl, or R c , R d Together with the nitrogen atom to which it is attached, it forms a 3- to 6-membered heterocyclic group;

[0015] Het is selected from 5- or 6-membered nitrogen aromatic group;

[0016] L is selected from a bond, C1-C6 alkylene, -C(=O)-, -NR a C(=O)-、-C(=O)-N(R a )-, -OC(=O)-, -C(=O)O-;

[0017] Ar is selected from 5-10 membered heterocyclic group, 5-10 membered heteroaryl, C6-C 10 Aryl, C3-C8 carbocyclic group; the heterocyclic group, heteroaryl group, aryl group, carbocyclic group are optionally substituted by one or more groups selected from the following: halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy;

[0018] n is selected from 1 or 2; m is selected from 1, 2, 3, 4, 5 or 6;

[0019] The condition is: when X is S=O, Het is When R1' is a C3-C8 carbocyclic group, L is not a bond.

[0020] On the other hand, the present invention provides a pharmaceutical composition comprising a compound represented by formula (I) or its stereoisomers, tautomers, solvates, prodrugs, isotope-labeled substances and pharmaceutically acceptable salts thereof, and a pharmaceutically acceptable carrier.

[0021] In another aspect, the present invention provides a compound represented by formula (I) or its stereoisomers, tautomers, solvates, prodrugs, isotope-labeled substances and pharmaceutically acceptable salts, or a pharmaceutical composition comprising the same, for use in the preparation of a medicament for preventing and / or treating a disease mediated by phosphodiesterase 4B.

[0022] Beneficial effects: Compared with the prior art, the compounds of the present invention not only have significantly better inhibitory activity of phosphodiesterase 4B and stronger target selectivity, but also have better therapeutic effects on idiopathic pulmonary fibrosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 The results of lung tissue pathology test on the effects of the compounds of the present invention on chronic pulmonary obstructive pulmonary disease in rats are shown.

[0025] Figure 2 The results of lung histopathological examination showing the effects of the compounds of the present invention on bleomycin-induced pulmonary fibrosis in rats. DETAILED DESCRIPTION

[0026] definition

[0027] As used in this specification, the following words and phrases are generally intended to have the meanings set forth below, unless the context in which they are used indicates otherwise.

[0028] As used herein, the term "alkyl" refers to a monovalent group of a straight or branched saturated hydrocarbon chain (typically having 1 to 6 carbon atoms, 1 to 4 carbon atoms, or 1 to 3 carbon atoms). This term is exemplified by groups such as methyl, ethyl, 1-propyl (n-propyl), 2-propyl (isopropyl), 1-butyl (n-butyl), 2-methyl-1-propyl (isobutyl), 2-butyl (sec-butyl), 2-methyl-2-propyl (tert-butyl), 1-pentyl (n-pentyl), 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl, 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl-3-pentyl, 2,3-dimethyl-2-butyl, 3,3-dimethyl-2-butyl, and the like.

[0029] As used herein, the term "alkylene" refers to a divalent radical of a straight or branched saturated hydrocarbon chain (typically having 1 to 6 carbon atoms, 1 to 4 carbon atoms, or 1 to 3 carbon atoms). The term is exemplified by radicals such as methylene, ethylene, propylene, butylene, pentylene, hexylene, and the like.

[0030] As used herein, the term "alkenyl" refers to a linear or branched unsaturated hydrocarbon chain monovalent group having 2 to 6 carbon atoms (more typically 2 to 4 carbon atoms, or 2 to 3 carbon atoms) and having a carbon-carbon double bond (e.g., 1 or 2 carbon-carbon double bonds). The term is exemplified by groups such as vinyl (i.e., -CH=CH2), propen-1-yl (i.e., -CH=CHCH3), propen-3-yl (or allyl, i.e., -CH2CH=CH2), propen-2-yl (i.e., -C(CH3)=CH2), butadienyl (including 1,2-butadienyl and 1,3-butadienyl), and the like.

[0031] As used herein, the term "alkynyl" refers to a linear or branched unsaturated hydrocarbon chain monovalent group (typically having 2 to 6 carbon atoms, 2 to 4 carbon atoms, or 2 to 3 carbon atoms) and having carbon-carbon triple bonds (e.g., 1 or 2 carbon-carbon triple bonds). The term is exemplified by groups such as ethynyl (i.e., -C≡CH), propargyl (i.e., -CH2C≡CH), propynyl (i.e., -C≡CCH3), etc.

[0032] As used herein, the term "aryl" refers to an aromatic carbocyclic group of 6 to 14 carbon atoms (more typically 6 to 10 carbon atoms, or 6 carbon atoms) having a single ring (e.g., phenyl) or multiple rings (e.g., biphenyl) or multiple condensed (fused) rings (e.g., naphthyl, fluorenyl, and anthracenyl). The term is exemplified by groups such as phenyl, fluorenyl, naphthyl, anthracenyl, 1,2,3,4-tetrahydronaphthalene (if the point of attachment is through the aryl group), and the like.

[0033] As used herein, the term "halogen" refers to fluorine, chlorine, bromine and iodine.

[0034] As used herein, the term "alkoxy" refers to an "alkyl-O-" group, wherein alkyl is as defined herein. This term is exemplified by groups such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, and the like.

[0035] As used herein, the term "carbocyclic radical" refers to a monoradical saturated or partially unsaturated group having 3 to 8 carbon atoms as 3 to 8 yuan of monocyclic rings or a plurality of thick (fused) rings or bridged rings or spirocycles.Carbocyclic ring or carbocyclic radical can be saturated or partially unsaturated, and can be fused with another saturated, partially unsaturated or aromatic ring, provided that the ring atom connected with the target molecule is not aromatic carbon.The example of carbocyclic ring or carbocyclic radical includes, but is not limited to cyclopropane, cyclobutane, cyclopentane, cyclohexane, cyclobutene, cyclopentene, cyclohexene, cycloheptene, cyclopentadiene etc.

[0036] As used herein, the term "heteroaryl" refers to an aromatic ring group containing a monocyclic or multiple condensed (fused) ring (e.g., containing 2 rings) of 5 to 10 ring atoms in the ring, wherein in addition to carbon atoms, the ring atoms also include at least one heteroatom selected from oxygen, nitrogen and / or sulfur. If the ring is aromatic, sulfur and nitrogen atoms can also exist in oxidized form. Multiple condensed (fused) ring heteroaryl is a monocyclic heteroaryl as defined above fused with one or more rings selected from the following to form a multiple condensed ring system: heteroaryl (to form, for example, naphthyridinyl, such as 1,8-naphthyridinyl), heterocycle (for example, 1,2,3,4-tetrahydronaphthyridinyl, such as 1,2,3,4-tetrahydro-1,8-naphthyridinyl), carbocycle (to form, for example, 5,6,7,8-tetrahydroquinolinyl) and aryl (to form, for example, indazolyl). It should be understood that the point of attachment of the heteroaryl can be on any suitable atom of the heteroaryl, including carbon atoms and heteroatoms (e.g., nitrogen). Exemplary heteroaryl groups include, but are not limited to, pyridinyl, pyrrolyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrazolyl, thienyl, indolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, furanyl, oxadiazolyl, thiadiazolyl, quinolinyl, isoquinolinyl, benzothiazolyl, benzoxazolyl, indazolyl, quinoxalinyl, quinazolinyl, 5,6,7,8-tetrahydroisoquinolinyl, benzofuranyl, benzimidazolyl, thiaindenyl, pyrrolo[2,3-b]pyridinyl, quinazolinyl-4(3H)-one, triazolyl, 4,5,6,7-tetrahydro-1H-indazolyl, and 3b,4,4a,5-tetrahydro-1H-cyclopropane[3,4]cyclopenta[1,2-c]pyrazolyl.

[0037] As used herein, the term "heterocyclyl" refers to a monovalent or divalent saturated or partially unsaturated group having 3 to 8-membered monocycles or multiple condensed (condensed) rings or bridged rings or spirocycles with 3 to 14 ring atoms in the ring, wherein in addition to carbon atoms, the ring atoms also include at least one or more nitrogen atoms. Examples of heterocyclyl groups include, but are not limited to, aziridine rings, azetidine rings, tetrahydropyrrole rings, piperidine rings, azepane rings, azocyclooctane rings, tetrahydroimidazole rings, tetrahydropyrazole rings, tetrahydrooxazole rings, tetrahydroisoxazole rings, tetrahydrothiazole rings, tetrahydroisothiazole rings, piperazine rings, morpholine rings, dihydropyridyl, 4,5,6,7-tetrahydro-1H-benzo[d]imidazole, 4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine, etc.

[0038] As used herein, the term "therapeutically effective amount" refers to an amount sufficient to affect treatment as defined below when administered to a mammal in need of such treatment. The therapeutically effective amount will vary with the subject and disease condition being treated, the weight and age of the subject, the severity of the disease condition, the mode of administration, etc., and can be readily determined by one of ordinary skill in the art.

[0039] As used herein, the term "stereoisomer" refers to a compound having the same chemical composition and connectivity, but whose atoms have different orientations in space that cannot be interchanged by single bond rotation. "Stereoisomer" includes "diastereomers" and "enantiomers". "Diastereomers" refers to stereoisomers with two or more chiral centers and whose molecules are not mirror images of each other. Diastereomers have different physical properties, such as melting points, boiling points, spectral characteristics and reactivity. Diastereomeric mixtures can be separated under high-resolution analytical procedures (such as crystallization, electrophoresis and chromatography). "Enantiomers" refers to two stereoisomers that are non-overlapping mirror images of each other.

[0040] As used herein, the term "tautomer" refers to the coexistence of two (or more) compounds that differ only in the position and electron distribution of one (or more) mobile atoms, such as keto-enol tautomers.

[0041] As used herein, the term "pharmaceutically acceptable salt" refers to salts that retain the biological effectiveness and properties of a given compound and are not biologically or otherwise undesirable. Pharmaceutically acceptable salts can be acid addition salts and / or base addition salts. Acid addition salts can be prepared from inorganic acids and organic acids. Salts derived from inorganic acids include hydrochlorides, hydrobromides, sulfates, nitrates, phosphates, carbonates, bisulfates, hydrogenphosphates, dihydrogenphosphates, bicarbonates, etc.; salts derived from organic acids include formates, acetates, propionates, glycolates, pyruvates, oxalates, malates, malonates, succinates, maleates, fumarates, tartrates, citrates, benzoates, cinnamates, mandelates, methanesulfonates, ethanesulfonates, p-toluenesulfonates, salicylates, lactates, nicotinates, lauryl sulfates, naphthylsulfonates, camphorsulfonates, gluconates, glucuronates, oleates, palmitates, stearates, pamoates, trifluoroacetates, etc. Base addition salts can be formed with inorganic or organic bases. Salts derived from inorganic bases include sodium, potassium, ammonium, calcium, magnesium, iron, zinc, copper, lithium, barium, aluminum salts and the like; salts derived from organic bases include salts formed with various primary, secondary and tertiary amines, for example, ethylamine, diethylamine, n-propylamine, isopropylamine, diethanolamine, meglumine, lysine, piperazine, piperidine, morpholine, tromethamine, choline and the like.

[0042] As used herein, the term "pharmaceutically acceptable" means that the substance or composition must be chemically and / or toxicologically compatible with the other ingredients comprising the formulation and / or the mammal to be treated therewith.

[0043] As used herein, the term "solvate" refers to an association or complex of one or more solvent molecules with a compound of the invention. Examples of solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, and ethanolamine. The term "hydrate" refers to a complex in which the solvent molecule is water.

[0044] As used herein, the term "prodrug" refers to those compounds that readily undergo chemical changes under physiological conditions to provide compounds of the present invention. In addition, prodrugs can be converted into compounds of the present invention by chemical or biochemical methods in an ex vivo environment. For example, when a prodrug is placed in a transdermal patch reservoir together with a suitable enzyme or chemical reagent, the prodrug can be slowly converted into a compound of the present invention.

[0045] Any formula or structure given herein, including Formula I or any formula disclosed herein, is also intended to represent unlabeled forms of the compound as well as isotopically labeled forms. These forms of the compound may also be referred to as "isotopically labeled." An isotopically labeled compound has a structure as depicted herein except that one or more atoms are replaced by an atom having a selected atomic mass or mass number. Examples of isotopes that may be incorporated into the compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, chlorine, and iodine, such as, but not limited to, 2 H (deuterium, D), 3 H (tritium), 11 C. 13 C. 14 C. 13 N. 15 N. 15 O. 17 O. 18 O. 31 P. 32 P. 35 S. 18 F. 36 Cl, 123 I and 125 I. Various isotope-labeled substances of the present invention, for example, substances in which radioactive isotopes (e.g. 3 H. 13 C and 14 C). Such compounds are synthesized by means well known in the art, for example by employing starting materials in which one or more hydrogens have been replaced with deuterium.

[0046] Compound

[0047] In one embodiment, the compound of the present invention is a compound represented by formula (I):

[0048]

[0049] or its stereoisomers, tautomers, solvates, prodrugs, isotope labels and pharmaceutically acceptable salts thereof, wherein X, R1, R1', R2, R2', R3, R3', Het, L, Ar, and n are as described above.

[0050] In one embodiment, the compound of the present invention is a compound represented by formula (I) or its stereoisomers, tautomers, solvates, prodrugs, isotope-labeled substances and pharmaceutically acceptable salts thereof, wherein:

[0051] X is selected from S=O, S(=O)2;

[0052] R1 is selected from H, C1-C6 alkyl;

[0053] R1' is selected from C3-C8 carbocyclic group, C6-C 10 aryl, 5-10 membered heteroaryl; wherein the carbocyclic group, aryl, heteroaryl is replaced by R4-(CR a R b ) m -substituted and optionally substituted by halogen, hydroxy, amino, mercapto, C1-C6 alkyl, C1-C6 alkoxy;

[0054] R2, R2', R3, R3' are each independently selected from hydrogen, C1-C6 alkyl;

[0055] R4 is selected from hydroxyl, amino, thiol, carboxyl, -SO3H, -PO4H, tetrazolyl, triazolyl, -C(=O)OC1-C6 alkyl, -C(=O)NR c R d ;

[0056] R a , R b are each independently selected from hydrogen, C1-C6 alkyl, halogen, hydroxyl, or R a , R b Together with the carbon atom to which it is attached, it forms a C3-C6 carbocyclic group; R c , R d are each independently selected from hydrogen, C1-C6 alkyl, or R c , R d Together with the nitrogen atom to which it is attached, it forms a 3- to 6-membered heterocyclic group;

[0057] Het is selected from 5- or 6-membered nitrogen aromatic group;

[0058] L is selected from a bond, C1-C6 alkylene, -C(=O)-, -NR e C(=O)-、-C(=O)-N(R e)-, -OC(=O)-, -C(=O)O-; R e Selected from hydrogen or C1-C6 alkyl;

[0059] Ar is selected from 5-10 membered heteroaryl, C6-C 10 Aryl; the heteroaryl and aryl are optionally substituted by one or more groups selected from the following: halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy;

[0060] n is selected from 1 or 2; m is selected from 1, 2, 3, 4, 5 or 6;

[0061] The condition is: when X is S=O, Het is When R1' is a C3-C8 carbocyclic group, L is not a bond.

[0062] In one embodiment, the compound of the present invention is a compound represented by formula (I) or its stereoisomers, tautomers, solvates, prodrugs, isotope-labeled substances, and pharmaceutically acceptable salts thereof, wherein

[0063] R1' is selected from cyclopropane, cyclobutane, cyclopentane, cyclohexane, phenyl, pyrrolyl, furanyl, thienyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl; the above groups are replaced by R4-(CR a R b ) m -substituted and optionally substituted by halogen, hydroxy, amino, mercapto, C1-C6 alkyl, C1-C6 alkoxy;

[0064] R2, R2', R3, R3' are each independently selected from hydrogen, C1-C6 alkyl;

[0065] R4 is selected from hydroxyl, amino, thiol, carboxyl, -SO3H, -PO4H, tetrazolyl, triazolyl, -C(=O)OC1-C 20 Alkyl, -C(=O)NR c R d ;

[0066] R a , R b are each independently selected from hydrogen, C1-C6 alkyl, halogen, hydroxyl, or R a , R b Together with the carbon atom to which it is attached, it forms a C3-C6 carbocyclic group; R c , R d are each independently selected from hydrogen, C1-C6 alkyl, or R c , R d Together with the nitrogen atom to which it is attached, it forms a 3- to 6-membered heterocyclic group;

[0067] Het is selected from 5- or 6-membered nitrogen aromatic group;

[0068] L is selected from a bond, C1-C6 alkylene, -C(=O)-, -NR a C(=O)-、-C(=O)-N(R a )-, -OC(=O)-, -C(=O)O-;

[0069] Ar is selected from pyrrolyl, furanyl, thienyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, phenyl; the above groups are optionally substituted by one or more groups selected from the following: halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy; n is selected from 1 or 2; m is selected from 1, 2, 3, 4, 5 or 6;

[0070] The condition is: when X is S=O, Het is When R1' is cyclopropane, cyclobutane, cyclopentane or cyclohexane, L is not a bond.

[0071] In one embodiment, the compound of the present invention is a compound represented by formula (I) or its stereoisomers, tautomers, solvates, prodrugs, isotope-labeled substances and pharmaceutically acceptable salts thereof, wherein

[0072] R1' is selected from cyclobutane, phenyl, pyridyl; the above groups are replaced by R4-(CR a R b ) m -substituted and optionally substituted by halogen;

[0073] R2, R2', R3, R3' are each independently selected from hydrogen, C1-C6 alkyl;

[0074] R4 is selected from hydroxyl, amino, thiol, carboxyl, -SO3H, -PO4H, tetrazolyl, triazolyl, -C(=O)OC1-C 20 Alkyl, -C(=O)NR c R d ;

[0075] R a , R b are each independently selected from hydrogen, C1-C3 alkyl, halogen, or R a , R b Together with the carbon atom to which it is connected, it forms a cyclopropane group or a cyclobutane group; R c , R d are each independently selected from hydrogen, C1-C3 alkyl, or R c , R dTogether with the nitrogen atom to which it is attached, it forms a 3- to 6-membered heterocyclic group;

[0076] Het is selected from 5- or 6-membered nitrogen aromatic group;

[0077] L is selected from a bond, C1-C6 alkylene, -C(=O)-, -NR a C(=O)-、-C(=O)-N(R a )-, -OC(=O)-, -C(=O)O-;

[0078] Ar is selected from pyrrolyl, furanyl, thienyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, phenyl; the above groups are optionally substituted by one or more groups selected from the following: halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy; n is selected from 1 or 2; m is selected from 1, 2, 3, 4, 5 or 6;

[0079] The condition is: when X is S=O, Het is When R1' is cyclobutane, L is not a bond.

[0080] In one embodiment, the compound of the present invention is a compound represented by formula (II):

[0081]

[0082] or its stereoisomers, tautomers, solvates, prodrugs, isotope-labeled substances, and pharmaceutically acceptable salts thereof, wherein:

[0083] X is selected from S=O, S(=O)2;

[0084] R1 is selected from H, C1-C6 alkyl;

[0085] L is selected from a bond, C1-C6 alkylene, -C(=O)-, -NR e C(=O)-、-C(=O)-N(R e )-, -OC(=O)-, -C(=O)O-; R e Selected from hydrogen or C1-C6 alkyl;

[0086] Ar is selected from pyrrolyl, furanyl, thienyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, phenyl; the above groups are optionally substituted by one or more groups selected from the following: halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy;

[0087] Provided that: when X is S=O, L is not a bond.

[0088] In one embodiment, the compound of the present invention is a compound represented by formula (III):

[0089]

[0090] or its stereoisomers, tautomers, solvates, prodrugs, isotope-labeled substances, and pharmaceutically acceptable salts thereof, wherein:

[0091] X is selected from S=O, S(=O)2;

[0092] R1 is selected from H, C1-C6 alkyl;

[0093] L is selected from a bond, C1-C6 alkylene, -C(=O)-, -NR e C(=O)-、-C(=O)-N(R e )-, -OC(=O)-, -C(=O)O-; R e Selected from hydrogen or C1-C6 alkyl;

[0094] Ar is selected from pyrrolyl, furanyl, thienyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, phenyl; the above groups are optionally substituted by one or more groups selected from the following: halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy.

[0095] In one embodiment, the compound of the present invention is a compound represented by formula (IV):

[0096]

[0097] or its stereoisomers, tautomers, solvates, prodrugs, isotope-labeled substances, and pharmaceutically acceptable salts thereof, wherein:

[0098] X is selected from S=O, S(=O)2;

[0099] R1 is selected from H, C1-C6 alkyl;

[0100] L is selected from a bond, C1-C6 alkylene, -C(=O)-, -NR e C(=O)-、-C(=O)-N(R e )-, -OC(=O)-, -C(=O)O-; R e Selected from hydrogen or C1-C6 alkyl;

[0101] Ar is selected from pyrrolyl, furanyl, thienyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, phenyl; the above groups are optionally substituted by one or more groups selected from the following: halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy;

[0102] Provided that: when X is S=O, L is not a bond.

[0103] In one embodiment, the compound of the present invention is a compound represented by formula (V):

[0104]

[0105] or its stereoisomers, tautomers, solvates, prodrugs, isotope-labeled substances, and pharmaceutically acceptable salts thereof, wherein:

[0106] X is selected from S=O, S(=O)2;

[0107] R1 is selected from H, C1-C6 alkyl;

[0108] R4 is selected from carboxyl, -SO3H, -PO4H, tetrazolyl, triazolyl, -C(=O)OC1-C6 alkyl, -C(=O)NR c R d ;

[0109] R a , R b are each independently selected from hydrogen, C1-C3 alkyl, halogen, or R a , R b Together with the carbon atom to which it is connected, it forms a cyclopropane group or a cyclobutane group; R c , R d are each independently selected from hydrogen, C1-C3 alkyl, or R c , R d Together with the nitrogen atom to which it is attached, it forms a 3- to 6-membered heterocyclic group;

[0110] R5 is selected from halogen, hydroxy, amino, thiol, C1-C6 alkyl, C1-C6 alkoxy;

[0111] Het is selected from 5- or 6-membered nitrogen aromatic group;

[0112] L is selected from a bond, C1-C6 alkylene, -C(=O)-, -NR e C(=O)-、-C(=O)-N(R e )-, -OC(=O)-, -C(=O)O-; R e Selected from hydrogen or C1-C6 alkyl;

[0113] Ar is selected from pyrrolyl, furanyl, thienyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, phenyl; the above groups are optionally substituted by one or more groups selected from the following: halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy;

[0114] p is selected from 1, 2 or 3; q is selected from 1, 2 or 3.

[0115] In one embodiment, the compound of the present invention is a compound represented by formula (V) or its stereoisomers, tautomers, solvates, prodrugs, isotope-labeled substances, and pharmaceutically acceptable salts thereof, wherein X is selected from S=O, S(=O)2;

[0116] R1 is selected from H, C1-C6 alkyl;

[0117] R4 is selected from carboxyl, -SO3H, -PO4H, tetrazolyl, triazolyl, -C(=O)OC1-C6 alkyl;

[0118] R a , R b are each independently selected from hydrogen, halogen, or R a , R b Together with the carbon atom to which it is attached, it forms a cyclopropane group or a cyclobutane group;

[0119] R5 is selected from halogen, hydroxy, amino, thiol, C1-C6 alkyl, C1-C6 alkoxy;

[0120] Het is selected from triazole;

[0121] L is selected from a bond, C1-C6 alkylene, -C(=O)-, -NR e C(=O)-、-C(=O)-N(R e )-, -OC(=O)-, -C(=O)O-; R e Selected from hydrogen or C1-C6 alkyl;

[0122] Ar is selected from pyridyl, pyrimidinyl, phenyl; the above groups are optionally substituted by one or more groups selected from the following: halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy;

[0123] p is selected from 1; q is selected from 1.

[0124] In one embodiment, compounds of the invention include, but are not limited to, the following compounds:

[0125]

[0126]

[0127] or a stereoisomer, a tautomer, a solvate, a prodrug, or a pharmaceutically acceptable salt thereof.

[0128] Pharmaceutical compositions and administration

[0129] The pharmaceutical composition provided by the present invention comprises the compound of the present invention or its stereoisomer, tautomer, solvate, prodrug, isotope label and pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are known to those skilled in the art, including diluents, lubricants, disintegrants, adhesives, buffers, preservatives, stabilizers, wetting agents, glidants, emulsifiers, colorants, flavoring agents, sweeteners, etc. Depending on the different routes of administration of the drug, such as oral administration, parenteral administration and rectal administration, the pharmaceutical composition of the present invention can be made in solid form (including but not limited to tablets, capsules, pills, granules, powders, powders, suppositories) or liquid form (including but not limited to solutions, suspensions, emulsions, tinctures, syrups). When the pharmaceutical composition of the present invention is in solid form, the pharmaceutically acceptable carrier generally includes one or more of the following: a) diluents, such as lactose, glucose, sucrose, mannitol, sorbitol, cellulose, etc.; b) lubricants, such as silicon dioxide, talc, stearic acid, polyethylene glycol, etc.; c) binders, such as magnesium aluminosilicate, gelatinized starch, gelatin, tragacanth gum, methylcellulose, sodium carboxymethylcellulose, microcrystalline cellulose, polyvinyl pyrrolidone, etc.; d) disintegrants, such as starch, alginic acid, agar, corn starch; e) stabilizers, such as antioxidants such as ascorbic acid; f) glidants, such as silicon dioxide; g) flavoring agents, such as mint, methyl salicylate; sweeteners, such as sucrose, saccharin. When the pharmaceutical composition of the present invention is in liquid form, the pharmaceutically acceptable carrier generally includes one or more of the following: a) a diluent, such as water for injection, physiological saline, Ringer's solution, polyethylene glycol, glycerol, propylene glycol, etc.; b) an antioxidant, such as ascorbic acid or sodium bisulfite; c) a buffer, such as acetate, phosphate, etc.

[0130] The effective dose of the compounds of the present invention depends at least on the nature, extent, delivery method and pharmaceutical dosage form of the condition being treated, and will be ultimately determined by the clinician. It can be expected that about 0.0001 to about 100 mg per kilogram of body weight per day; generally about 0.01 to about 10 mg per kilogram of body weight per day; more typically, about 0.01 to about 5 mg per kilogram of body weight per day; most typically about 0.05 to about 0.5 mg per kilogram of body weight per day. For example, the daily candidate dose for an adult of about 70 kg body weight will be in the range of 1 mg to 1000 mg, preferably in the range of 5 mg to 500 mg, and can be administered in the form of a single dose or multiple doses.

[0131] Indications

[0132] The compounds of the present invention have extremely strong inhibitory activity on PDE4B and can be used to prevent and / or treat inflammatory diseases associated with PDE4B, including but not limited to atopic dermatitis, asthma, chronic obstructive pulmonary disease (COPD), idiopathic pulmonary fibrosis, interstitial lung disease, chronic sinusitis, allergic rhinitis, allergic dermatitis, contact dermatitis, psoriasis, systemic lupus erythematosus, ulcerative colitis, Crohn's disease, depression, bipolar depression, mania, anxiety, schizophrenia, Alzheimer's disease, stroke, chronic pain, liver fibrosis, renal fibrosis, and nephritis.

[0133] Example

[0134] Example 1

[0135] Step 1 Synthesis of tert-butyl 4-(5-chloropyrimidin-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate (2)

[0136]

[0137] To a 100 mL single-necked bottle, tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate (1,1.00 g, 3.23 mmol), 5-chloro-2-iodopyrimidine (932 mg, 3.88 mmol), dioxane (10 mL), H2O (2 mL) and Na2CO3 (1.02 g, 9.70 mmol) were added in sequence, and the atmosphere was replaced with nitrogen three times. Finally, Pd(PPh3)4 (186 mg, 0.16 mmol) was added and the reaction was carried out at 90°C for 16 h. TLC (V 石油醚 :V 乙酸乙酯 =10:1) to monitor the complete reaction of the raw materials. Water (50 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (50 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated and purified by column chromatography (SiO2, V 石油醚 :V 乙酸乙酯 =20:1) to obtain 510 mg of white solid 2 with a yield of 53.4%. 1 H NMR (400MHz, CDCl3) δ (ppm): 8.62 (s, 2H, ArH), 7.19 (s, 1H, CH), 4.17 (m, 2H, CHCH2), 3.63 (m, 2H, NCH2), 2.68 (m, 2H, CCH2), 1.49 (s, 9H, C(CH3)3). Step 2 Synthesis of 5-chloro-2-(1,2,3,6-tetrahydropyridin-4-yl)pyrimidine hydrochloride (3)

[0138]

[0139] 2 (510 mg, 1.72 mmol), anhydrous dichloromethane (2 mL) and HCl (4 M dioxane solution, 6 mL) solution were added to a 50 mL single-necked bottle in sequence. The reaction was carried out at room temperature for 2 h under nitrogen protection. TLC (V 石油醚 :V 乙酸乙酯 =10:1) The reaction of the raw material was monitored to be complete. The reaction solution was directly concentrated to obtain 390 mg of white solid 3 crude product, which was used directly in the next step without purification.

[0140] 1 H NMR(400MHz,DMSO-d6)δ(ppm):9.03(s,2H,NH,HCl),8.95(s,2H,ArH),7.16(s,1H,C =CH), 3.86 (d, J = 4.4Hz, 2H, CHCH2), 3.38 (m, 2H, NHCH2), 2.77 (d, J = 2.1Hz, 2H, CCH2).

[0141] Step 3 Synthesis of (1-aminocyclobutyl)methanol hydrochloride (5)

[0142]

[0143] To a 50mL Schlenk reaction tube, tert-butyl (1-(hydroxymethyl)cyclobutyl)carbamate (4, 3.00g, 14.90mmol) and dichloromethane (6mL) were added in sequence. After stirring to dissolve, a 1,4-dioxane solution of hydrogen chloride (4.0M, 6mL) was added dropwise to the reaction system at 0°C. After the addition was completed, the reaction was carried out at room temperature under nitrogen protection for 3h. TLC (V 二氯甲烷 :V 甲醇 =50:1) monitor the reaction of the raw materials to complete. The reaction solution was concentrated under reduced pressure, and the obtained crude product was slurried with ethyl acetate (5 mL), filtered, and dried to obtain 1.98 g of white solid 5, with a yield of 96.6%.

[0144] 1 H NMR (600MHz, DMSO-d6) δ (ppm): 8.20 (br, 3H, NH2, HCl), 5.47 (t, J = 5.3Hz, 1H, OH), 3.55 (d, J = 5.3 Hz,2H,CH2OH),2.23-2.11(m,2H,CH2CH2CH2),2.05-1.96(m,2H,CH2CH2CH2),1.90-1.81(m,1H,H of CH2CH2CH2),1.79-1.71(m,1H,Hof CH2CH2CH2); HRMS(ESI):m / z[M-HCl+H]+ Theoretical value C5H 11 NO:102.0919; measured value:102.0915.

[0145] Step 4 Synthesis of (1-((2-chloro-6,7-dihydrothieno[3,2-d]pyrimidin-4-yl)amino)cyclobutyl)methanol (7)

[0146]

[0147] 2,4-Dichloro-6,7-dihydrothieno[3,2-d]pyrimidine (6, 1.37 g, 6.60 mmol), 5 (1.00 g, 7.27 mmol), acetonitrile (8 mL) and triethylamine (3.34 g, 33.00 mmol) were added to a 50 mL Schlenk solution in sequence, and the reaction was carried out at 76°C under nitrogen protection overnight. TLC (V 石油醚 :V 乙酸乙酯 =1:1) The reaction was monitored to be complete, and about 30% of the raw material remained. The reaction solution was concentrated under reduced pressure and purified by rapid preparative chromatography (20 g, V 石油醚 :V 乙酸乙酯 =1:1) to obtain 1.08 g of white solid 7 with a yield of 60.0%.

[0148] 1 H NMR (600MHz, CDCl3) δ (ppm): 4.67 (s, 1H, NH), 3.90 (s, 3H, CH2OH), 3.44-3.39 (m, 2H, SCH2), 3.31-3.2 3(m,2H,SCH2CH2),2.41-2.32(m,2H,CH2CH2CH2),2.19-2.11(m,2H,CH2CH2CH2),2.03-1.94(m,1H,H of CH2CH2CH2),1.94-1.85(m,1H,H of CH2CH2CH2); HRMS(ESI):m / z[M+H] + Theoretical value C 11 H 14 ClN3OS:272.0624; Measured value:272.0604.

[0149] Step 5 Synthesis of 2-chloro-4-((1-(hydroxymethyl)cyclobutyl)amino)-6,7-dihydrothieno[3,2-d]pyrimidine-5-oxide (8)

[0150]

[0151] 7 (500 mg, 1.84 mmol), S-(-)-BINOL (53 mg, 0.184 mmol), dichloromethane (3 mL), tetraisopropyl titanate (26 mg, 0.092 mmol) and water (33 mg, 1.84 mmol) were added to a 50 mL Schlenk reaction tube in sequence. After the addition was completed, the mixture was stirred at room temperature under nitrogen protection for 1 h. Subsequently, tert-butyl peroxide (262 mg, 2.03 mmol) was added. After the addition was completed, the mixture was reacted at room temperature under nitrogen protection for 2 h. TLC (V 二氯甲烷 :V 甲醇 =15:1) to monitor the complete reaction of the raw materials. The system was directly purified by column chromatography without treatment (SiO2, V 二氯甲烷 :V 甲醇 =15:1) to obtain 340 mg of light yellow solid 8, with a yield of 64.2%.

[0152] 1 H NMR(600MHz,DMSO-d6)δ(ppm):8.61(s,1H,NH),4.90(t,J=5.6Hz,1H,OH),3.75-3.65(m,2H,CH2OH),3.60-3.52(m,1H,H of SCH2),3.39-3.34(m,1H,Hof SCH2),3.18-3.10(m,1H,H of SCH2CH2),3.06-2.99(m,1H,H of SCH2CH2),2.31-2.15(m,4H,CH2CH2CH2),1.83-1.69(m,2H,CH2CH2CH2); HRMS(ESI):m / z[M+H] + Theoretical value C 11 H 14 ClN3O2S: 288.0573; Measured value: 288.0554.

[0153] Step 6 Preparation of 2-(4-(5-chloropyrimidin-2-yl)-3,6-dihydropyridin-1(2H)-yl)-4-((1-(hydroxymethyl)cyclobutyl)amino)-6,7-dihydrothiophene[3,2-d]pyrimidine-5-oxide (Example 1)

[0154]

[0155] 3 (168 mg, 0.72 mmol), 8 (200 mg, 0.69 mmol), N,N-diisopropylethylamine (224 mg, 1.73 mmol), tetrahydrofuran (8 mL) and water (2 mL) were added to a 50 mL single-necked bottle in sequence. After the addition, the atmosphere was replaced with nitrogen three times and the reaction was carried out at 65 °C overnight. TLC (V 二氯甲烷 :V 甲醇=10:1) to monitor the complete reaction of the raw materials. Add water (20 mL) to the reaction solution, extract with ethyl acetate (20 mL×3), combine the organic phases, dry with anhydrous sodium sulfate, filter, concentrate and purify through a thick preparation plate (V 二氯甲烷 :V 甲醇 =10:1) to obtain 104 mg of the compound of Example 1 as a white solid with a yield of 44.6%.

[0156] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 8.88 (s, 2H, ArH), 7.37 (s, 1H, C=CH), 7.28 (s, 1H, NH), 4.84 (t, J=5.7Hz, 1H, OH),4.45(s,2H,CH2OH),3.96(t,J=5.6Hz,2H,NCH2CH2),3.76-3.69(m,2H,NCH2CH),3.41(d,J=8.5Hz,1H,H of SCH2),3.22-3.14(m,1H,H ofSCH2),2.98-2.90(m,1H,H ofSCH2CH2),2.89-2.82(m,1H,H of SCH2CH2),2.64(s,2H,NCH2CH2),2.35(m,2H,H of CH2CH2CH2),2.25-2.16(m,2H,H of CH2CH2CH2),1.78(d,J=9.5Hz,2H,H of CH2CH2CH2); 13 C NMR (100MHz, DMSO-d6) δ (ppm): 175.32, 162.09, 158.12, 156.22, 128.79, 58.97, 48.94, 29.47, 14.41.; HRMS (ESI): m / z [M+H] + Theoretical value C 20 H 23 ClN6O2S:447.1370; Measured value:447.1363

[0157] Example 2

[0158] Step 1 Synthesis of 2-bromomethyl-5-chloropyrimidine (2)

[0159]

[0160] Compound 1 (1.0 g, 7.78 mmol) and carbon tetrachloride (20 mL) were added to a 50 mL single-necked bottle in sequence. NBS (1.52 g, 8.56 mmol) and BPO (0.19 g, 0.78 mmol) were added under stirring. After the addition was completed, the mixture was refluxed under nitrogen protection for 36 h. TLC (V 石油醚 :V 乙酸乙酯 =10:1) monitor the remaining 30% of the raw material. The reaction solution was cooled to room temperature, filtered, the filter cake was washed with dichloromethane (10 mL), the filtrate was concentrated and purified by flash preparative chromatography (12 g, V 石油醚 :V 乙酸乙酯 =10:1) to obtain 320 mg of white solid intermediate 2 with a yield of 17.8%.

[0161] 1 H NMR (600 MHz, CDCl3) δ (ppm): 8.68 (s, 2H, ArH), 4.58 (s, 2H, CH2). Step 2 Synthesis of tert-butyl 4-((5-chloropyrimidin-2-yl)methyl)piperazine-1-carboxylate (3)

[0162]

[0163] To a 50 mL single-mouth bottle, add intermediate 2 (0.35 g, 1.69 mmol), N-Boc-piperazine (0.38 g, 2.03 mmol) and dichloromethane (20 mL) in sequence, cool to 0 ° C, add triethylamine (0.21 g, 2.03 mmol), and react at room temperature for 3 h under nitrogen protection. TLC (ethyl acetate) monitors the completion of the reaction. Add water (1.5 mL) to quench the reaction, extract the reaction solution with dichloromethane (30 mL × 3), combine the organic phases, wash the organic phases with saturated sodium chloride solution (50 mL × 3), dry over anhydrous sodium sulfate, concentrate and purify by rapid preparative chromatography (12 g, V 石油醚 :V 乙酸乙酯 =1:1) to obtain 281 mg of white solid intermediate 3 with a yield of 53.3%.

[0164] 1 H NMR (600 MHz, DMSO-d6) δ (ppm): 8.86 (s, 2H, ArH), 3.70 (s, 2H, CH2), 3.27-3.23 (m, 4H, piperazine-H), 2.41-2.39 (m, 4H, piperazine-H), 1.34 (s, 9H, C(CH3)3).

[0165] Step 3 Synthesis of 1-((5-chloropyrimidin-2-yl)methyl)piperazine trifluoroacetate (4)

[0166]

[0167] Add intermediate 3 (0.28 g, 0.9 mmol) and dichloromethane (6 mL) to a 25 mL single-mouth bottle, cool to 0 ° C, add TFA (2 mL), and react at room temperature for 1 h under nitrogen protection. TLC (ethyl acetate) monitors the completion of the reaction. Concentrate to obtain 190 mg of white solid intermediate 4, with a yield of 99%.

[0168] Step 4 Synthesis of 2-(4-((5-chloropyrimidin-2-yl)methyl)piperazin-1-yl)-4-((1-(hydroxymethyl)cyclobutyl)amino)-6,7-dihydrothieno[3,2-d]pyrimidine 5-oxide (Example 2)

[0169]

[0170] To a 25 mL single-necked bottle, intermediate 4 (0.095 g, 0.45 mmol), intermediate 8 of Example 1 (0.129 g, 0.45 mmol), DIPEA (0.174 g, 1.35 mmol), THF (4 mL) and H2O (0.5 mL) were added in sequence. After the addition was completed, the reaction was carried out at 65°C under nitrogen protection overnight. TLC (V 二氯甲烷 :V 甲醇 =20:1) monitor the completion of the reaction, concentrate under reduced pressure, add THF (4 mL), slurry for 1 h, filter, and dry the filter cake to obtain 100 mg of white solid Example 2, with a yield of 47.9%.

[0171] 1 H NMR (400MHz, CDCl3) δ (ppm): 8.68 (s, 2H, ArH), 5.90 (s, 1H, NH), 4.13 (s, 1H, OH), 3.9 1-3.84(m,4H,Piperazine-H),3.84-3.82(m,4H,Piperazine-H),3.61-3.50(m,1H,H ofCH2),3.41-3.32(m,1H,H of CH2),3.05-2.91(m,2H,CH2),2.58(t,J=4.9Hz,4H,CH2,CH2),2.29-2.21(m,4H,CH2CH2),1.97-1.89(m,1H,H of CH2),1.87-1.82(m,1H,H of CH2).

[0172] 13C NMR(151MHz,DMSO-d6)δ(ppm):175.50,165.42,162.37,158.18,156.13,129.73 ,109.85,64.11,63.86,59.01,52.83,48.97,44.21,32.96,30.19,30.03,14.90.

[0173] MS (ESI): m / z [M+H] + Theoretical value C 20 H 26 ClN7O2S: 464.1635; Measured value: 464.1625. Example 3

[0174] Step 1 Synthesis of tert-butyl 4-(5-chloropyrimidine-2-carbonyl)piperazine-1-carboxylate (2)

[0175]

[0176] Compound 1 (500 mg, 3.15 mmol), DMF (10 mL), HATU (1.37 g, 3.6 mmol) and DIPEA (0.84 mL, 4.8 mmol) were added to a 100 mL single-necked bottle in sequence, and stirred at room temperature for 0.5 h under nitrogen protection. Then, tert-butyl piperazine-1-carboxylate (447 mg, 2.4 mmol) was added and the reaction was allowed to proceed overnight at room temperature. TLC (V 二氯甲烷 :V 甲醇 =20:1) to monitor the complete reaction of the raw material. Add water (30 mL) to quench, extract with ethyl acetate (30 mL×3), combine the organic phases, dry over anhydrous sodium sulfate, filter, concentrate, and purify by column chromatography (V 二氯甲烷 :V 甲醇 =15:1), to obtain 370 mg of yellow solid intermediate 2, with a yield of 47.0%.

[0177] 1 H NMR (600 MHz, DMSO-d6) δ (ppm): 9.04 (s, 2H, ArH), 3.65-3.61 (m, 2H, piperazine-CH2), 3.44-3.41 (m, 2H, piperazine-CH2), 3.29-3.25 (m, 2H, piperazine-CH2), 3.20-3.17 (m, 2H, piperazine-CH2), 1.40 (s, 9H, C(CH3)3).

[0178] Step 2 Synthesis of (5-chloropyrimidin-2-yl)(piperazine-1-yl)methanone hydrochloride (3)

[0179]

[0180] Compound 2 (320 mg, 0.98 mmol) and 1M HCl / dioxane (6 mL) solution were added to a 50 mL single-necked bottle, and the mixture was reacted at room temperature for 1 h under nitrogen protection. TLC (V 二氯甲烷 :V 甲醇 =20:1) Monitor the reaction of the raw materials to be complete. The reaction solution was directly concentrated to dryness under reduced pressure, and ethyl acetate (30 mL) was added for pulping and filtration to obtain 150 mg of yellow solid 3, with a yield of 58.2%.

[0181] 1 H NMR (400 MHz, D2O) δ (ppm): 8.88 (s, 2H, ArH), 3.95 (t, J = 5.6 Hz, 2H, piperazine-CH2), 3.66 (t, J = 5.2 Hz, 2H, piperazine-CH2), 3.35 (t, J = 5.6 Hz, 2H, piperazine-CH2), 3.23 (t, J = 5.2 Hz, 2H, piperazine-CH2).

[0182] Step 3 Synthesis of (5-chloropyrimidin-2-yl)(4-(4-((1-(hydroxymethyl)cyclobutyl)amino)-5-oxo-6,7-dihydrothieno[3,2-d]pyrimidin-2-yl)piperazine-1-yl)methanone (Example 3)

[0183]

[0184] To a 50 mL single-necked bottle, intermediate 3 (150 mg, 0.57 mmol), intermediate 8 (156 mg, 0.54 mmol) of Example 1, tetrahydrofuran (8 mL), DIPEA (0.24 mL, 1.35 mmol) and water (2 mL) were added in sequence, and the mixture was reacted at 65° C. overnight under nitrogen protection. TLC (V 二氯甲烷 :V 甲醇 =20:1) to monitor the complete reaction of the raw materials. Water (10 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (20 mL×3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated and purified by flash preparative chromatography (4 g, V 二氯甲烷 :V 甲醇 =20:1), and 200 mg of crude product was obtained. The crude product was slurried with tetrahydrofuran (10 mL) to obtain 170 mg of white solid Example 3, with a yield of 65.9%

[0185] 1H NMR (600 MHz, CDCl3) δ (ppm): 8.79 (s, 2H, ArH), 6.29 (s, 1H, NH), 4.07-3.93 (m, 3H, piperazine-H), 3.89-3.81 (m, 5H, piperazine-H), 3.78-3.72 (m, 1H, OH), 3.62-3.53 (m, 1H, H of SOCH2), 3.43-3.33 (m, 3H, 1H of SOCH2, 2H of SOCH2CH2), 3.08-2.94 (m, 2H, CH2OH), 2.37-2.24 (m, 2H, NHCCH2), 2.23-2.14 (m, 2H, NHCCH2), 1.93-1.81 (m, 2H, NHCCH2CH2).

[0186] 13 C NMR(150MHz,DMSO-d6)δ(ppm):175.50,164.52,162.37,159.99,158.19,156.84,131.29 ,110.54,64.08,59.06,49.05,46.32,44.60,43.83,41.56,33.03,30.21,30.05,14.92.

[0187] HRMS(ESI):m / z[M+H] + Theoretical value C 20 H 24 ClN7O3S: 478.1428; Measured value: 478.1399. Example 4

[0188] Step 1 Synthesis of 2-chloro-4-((1-(hydroxymethyl)cyclobutyl)amino)-6,7-dihydrothieno[3,2-d]pyrimidine 5,5-dioxide (2)

[0189]

[0190] In a 25 mL single-necked bottle, intermediate 7 (100 mg, 0.37 mmol), m-chloroperbenzoic acid (85%, 189 mg, 0.93 mmol) and dichloromethane (4 mL) were added in sequence. After the addition was complete, the mixture was reacted at room temperature for 7 h. TLC (V 石油醚 :V 乙酸乙酯 =1:1) to monitor the reaction of the raw materials. Add dichloromethane (10 mL) to the reaction solution, dilute, wash with saturated sodium thiosulfate solution (10 mL), saturated sodium bicarbonate solution (10 mL) and saturated brine (10 mL) in sequence, dry over anhydrous sodium sulfate, filter, concentrate and purify by flash preparative chromatography (4 g, V 石油醚 :V 乙酸乙酯=1:1), to obtain 79 mg of white solid intermediate 2, with a yield of 70.3%.

[0191] 1 H NMR (600MHz, DMSO-d6) δ (ppm): 7.53 (s, 1H, NH), 4.99 (t, J = 5.6Hz, 1H, OH), 3.67 (d, J = 5.7Hz, 2H, CH2OH), 3.63 (t, J = 6.9Hz, 2H, S CH2),3.26-3.20(m,2H,SCH2CH2),2.40-2.30(m,2H,CH2CH2CH2),2.20-2.12(m,2H,CH2CH2CH2),1.86-1.69(m,2H,CH2CH2CH2).

[0192] HRMS(ESI):m / z[M+H] + Theoretical value C 11 H 14 ClN3O3S: 304.0522; Found: 304.0503. Step 2 Synthesis of 2-(4-(5-chloropyrimidin-2-yl)piperidin-1-yl)-4-((6-(hydroxymethyl)bicyclo[3.2.0]heptane-6-yl)amino)-6,7-dihydrothieno[3,2-d]pyrimidine 5,5-dioxide (Example 4)

[0193]

[0194] To a 10 mL Schlenk reaction tube, intermediate 2 (150 mg, 0.494 mmol), 5-chloro-2-(piperidin-4-yl)pyrimidine (103 mg, 0.519 mmol), tetrahydrofuran (2 mL), water (0.5 mL) and N,N-diisopropylethylamine (160 mg, 1.24 mmol) were added in sequence. After the addition was completed, the reaction was carried out at 65 °C under nitrogen protection overnight. TLC (V 石油醚 :V 乙酸乙酯 =1:2) monitor the reaction of the raw materials to complete. The reaction system was filtered, and the filter cake was washed with tetrahydrofuran (5 mL), and dried to obtain 167 mg of white solid Example 4, with a yield of 72.7%.

[0195] 1H NMR (600MHz, CDCl3) δ (ppm): 8.62 (s, 2H, ArH), 5.61 (s, 1H, NH), 4.48-5.10 (m, 2H, OH and 1H of SCH2), 3.90 (d, J = 5.2Hz, 2H, CH2OH), 3.85 (t, J = 5.4Hz, 1H, H of SCH2),3.42(t,J=7.0Hz,2H,SCH2CH2),3.22-3.14(m,1H,CH),3.14-3.04(m,4H,N(CH2)2),2.35- 2.28(m,2H,CHCH2),2.28-2.20(m,2H,CHCH2),2.11-2.03(m,2H,CH2CH2CH2),2.02-1.94(m,1H,H of CH2CH2CH2),1.93-1.78(m,3H,CH2CH2CH2 and 1H of CH2CH2CH2).

[0196] 13 C NMR (151MHz, DMSO-d6) δ (ppm): 170.71, 168.62, 162.49, 156.20, 155.63, 129.06, 104.65, 64.68, 58.78, 50.53, 44.42, 30.75, 29.94, 28.84, 14.46.

[0197] HRMS(ESI):m / z[M+H] + Theoretical value C 20 H 25 ClN6O3S: 465.1475; Measured value: 465.1457. Example 5

[0198] Step 1 Synthesis of methyl 4-((2-methoxy-2-oxoethyl)thio)butanoate (3)

[0199]

[0200] Compound 2 (13.36 g, 0.126 mol) and methanol (90 mL) were added to a 500 mL single-necked bottle in sequence. After stirring at room temperature for 0.5 h, potassium iodide (150 mg, 0.88 mmol) and compound 1 (20.00 g, 0.146 mol) were added and stirred at 65°C for 20 h. TLC (V 石油醚 :V 乙酸乙酯=10:1) monitor the reaction of the raw materials, cool the reaction solution to room temperature, filter, collect and concentrate the filtrate, add DCM (200 mL) to dissolve, wash with saturated brine (100 mL×3), dry, filter and concentrate to obtain 26.00 g of light yellow oily intermediate 3, with a yield of 93.1%.

[0201] 1 H NMR (600MHz, CDCl3) δ (ppm): δ3.72(s,3H,OCH3),3.66(s,3H,OCH3),3.21(s,2H,COCH2S),2 .67(t,J=7.2Hz,2H,SCH2),2.43(t,J=7.2Hz,2H,COCH2),1.95-1.90(m,2H,COCH2SCH2CH2).

[0202] Step 2 Synthesis of methyl 3-oxotetrahydro-2H-thiopyran-2-carboxylate (4)

[0203]

[0204] Sodium methoxide (7.50 g, 0.139 mol), toluene (260 mL) and intermediate 3 (26.00 g, 0.126 mol) were added to a 500 mL single-necked bottle in sequence, and stirred at 105° C. for 3 h under a nitrogen atmosphere. TLC (V 石油醚 :V 乙酸乙酯 =10:1) monitor the reaction of the raw materials to complete, cool the reaction solution to ice bath and add concentrated hydrochloric acid (35 mL), add EA (150 mL × 3) to extract, combine the organic phases, wash with saturated brine (100 mL × 3), dry, filter, concentrate and purify by flash preparative chromatography (120 g × 2, V 石油醚 :V 乙酸乙酯 =100:3), to obtain 15.10 g of light yellow oily intermediate 4, with a yield of 86.7%.

[0205] Step 3 Synthesis of 2-(methylthio)-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-ol (6)

[0206]

[0207] Potassium hydroxide (6.30 g, 0.113 mol), methanol (85 mL), S-methylisothiourea sulfate (5) (12.80 g, 0.046 mol) and intermediate 4 (12.00 g, 0.069 mol) were added to a 250 mL single-necked bottle in sequence, and the mixture was stirred at room temperature for 16 h under a nitrogen atmosphere. TLC (V 二氯甲烷 :V 甲醇=15:1) Monitor the reaction of the raw materials to be complete, pour the reaction solution into ice water (150 mL), add acetic acid (20 mL), stir in an ice bath for 0.5 h, filter, collect the filter cake, stir with pure water (150 mL) at room temperature for 1 h, filter, wash the filter cake with water (30 mL×2), collect the filter cake and dry it to obtain 11.20 g of white solid intermediate 6, with a yield of 76.2%.

[0208] 1 H NMR (600MHz, DMSO-d6) δ (ppm): 2.91-2.87 (m, 2H, SCH2), 2.61 (t, J = 6.3Hz, 2H, ArCH2), 2.45 (s, 3H, SCH3), 2.04-1.99 (m, 2H, SCH2CH2).

[0209] Step 4 Synthesis of 7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidine-2,4-diol (7)

[0210]

[0211] Intermediate 6 (8.00 g, 37.41 mmol), water (30 mL) and acetic acid (60 mL) were added to a 250 mL single-necked bottle in sequence, and stirred at 110° C. for 3 days under a nitrogen atmosphere. TLC (V 二氯甲烷 :V 甲醇 =10:1) monitor the reaction of the raw materials to complete, cool the reaction solution in an ice bath until solid precipitates, filter, wash the filter cake with water (30 mL×2), collect the filter cake and dry to obtain 5.60 g of white solid intermediate 7, with a yield of 82.3%.

[0212] 1 H NMR (600MHz, DMSO-d6) δ (ppm): 11.16 (s, 1H, OH), 10.82 (s, 1H, OH), 2.93-2.74 (m, 2H, SCH2), 2.46-2.35 (m, 2H, ArCH2), 1.99-1.96 (m, 2H, SCH2CH2). Step 5 Synthesis of 2,4-dichloro-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidine (8)

[0213]

[0214] To a 250 mL single-necked bottle, intermediate 7 (2.60 g, 14.20 mmol), phosphorus oxychloride (6 mL) and N,N-dimethylaniline (172 mg, 1.42 mmol) were added in sequence, and the mixture was stirred at 110° C. for 3 h under a nitrogen atmosphere. TLC (V 二氯甲烷 :V 甲醇=15:1) monitor the reaction of the raw materials to complete, cool to room temperature, add the reaction solution dropwise into ice water (100 mL) to quench, add DCM (100 mL×3) to extract, add saturated brine (50 mL×3) to wash, dry, filter, concentrate and purify by flash preparative chromatography (40 g, V 石油醚 :V 乙酸乙酯 =10:1), to obtain 2.50 g of white solid intermediate 8, with a yield of 80.6%.

[0215] 1 H NMR (600MHz, DMSO-d6) δ (ppm): 3.15-3.11 (m, 2H, SCH2), 2.90 (t, J = 6.2Hz, 2H, ArCH2), 2.12-2.05 (m, 2H, SCH2CH2).

[0216] Step 6 Synthesis of (1-((2-chloro-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)cyclobutyl)methanol (9)

[0217]

[0218] To a 50 mL single-necked bottle, intermediate 8 (584 mg, 2.64 mmol), acetonitrile (8 mL), intermediate 5 of Example 1 (400 mg, 2.91 mmol) and triethylamine (1.34 g, 16.52 mmol) were added in sequence and stirred at 75°C for 16 h under a nitrogen atmosphere. TLC (V 石油醚 :V 乙酸乙酯 =3:1) to monitor the reaction completion of the raw material, concentrate under reduced pressure and purify by flash preparative chromatography (24 g, V 石油醚 :V 乙酸乙酯 =3:2), to obtain 420 mg of white solid intermediate 9, with a yield of 55.9%.

[0219] 1 H NMR (600MHz, DMSO-d6) δ (ppm): 6.16 (s, 1H, NH), 4.86 (t, J = 5.6Hz, 1H, OH), 3.65 (d, J = 5.7Hz, 2H, HOCH2), 3.09-3.04 (m, 2 H,SCH2),2.67(t,J=6.3Hz,2H,ArCH2),2.31-2.26(m,2H,SCH2CH2),2.15-2.05(m,4H,CH2CH2),1.84-1.68(m,2H,CH2).

[0220] Step 7 Synthesis of 2-chloro-4-((1-(hydroxymethyl)cyclobutyl)amino)-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidine 5,5-dioxide (10)

[0221]

[0222] To a 50 mL single-necked bottle, intermediate 9 (300 mg, 1.05 mmol), DCM (15 mL) and 85% m-CPBA (532 mg, 2.62 mmol) were added in sequence, and the mixture was stirred at room temperature for 4 h under a nitrogen atmosphere. TLC (V 石油醚 :V 乙酸乙酯 =1:1) to monitor the reaction of the raw material, add sodium thiosulfate aqueous solution (30 mL) to quench, extract with DCM (40 mL×3), combine the organic phases, wash with saturated NaHCO3 solution (20 mL×2), saturated brine (30 mL) in sequence, dry, filter, concentrate and purify by flash preparative chromatography (12 g, V 二氯甲烷 :V 甲醇 =50:1), to obtain 320 mg of white solid intermediate 10, with a yield of 90.9%.

[0223] HRMS(ESI):m / z[M+H] + Theoretical value C 12 H 16 ClN3O3S: 3187.0684; Found: 318.0663. Step 8 Synthesis of 2-(4-(5-chloropyrimidin-2-yl)piperidin-1-yl)-4-((1-(hydroxymethyl)cyclobutyl)amino)-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidine 5,5-dioxide (Example 5)

[0224]

[0225] Compound 10 (315 mg, 0.99 mmol), THF (4 mL), water (0.5 mL), DIPEA (320 mg, 2.48 mmol) and 5-chloro-2-(piperidin-4-yl)pyrimidine (236 mg, 1.19 mmol) were added to a 25 mL single-necked bottle in sequence. The mixture was heated at room temperature at 65°C overnight under a nitrogen atmosphere. TLC (V 二氯甲烷 :V 乙酸乙酯 =1:1) to monitor the reaction completion of the raw material, add water (20 mL) to quench, extract with ethyl acetate (50 mL × 3), wash with saturated brine (20 mL × 2), dry, filter, concentrate and purify by flash preparative chromatography (12 g, V 二氯甲烷 :V 甲醇 =50:1) to obtain a crude product, which was slurried with a mixed solvent (DCM:MeOH=10:1, 15 mL) overnight, filtered, and the filter cake was washed with water (10 mL). The filter cake was collected and dried to obtain 410 mg of white solid Example 5, with a yield of 90.5%.

[0226] 1 H NMR (600MHz, CDCl3) δ (ppm): 8.61 (s, 2H, ArH), 6.81 (s, 1H, NH), 4.74 (br, 2H, SO2CH2), 4.12 (t, J=5.2Hz,1H,OH),3.88(d,J=5.1Hz,2H,HOCH2),3.34-3.26(m,2H,ArCH2),3.19-3.11(m,1H,CH) ,3.07-2.96(m,2H,NCH2),2.76(t,J=6.5Hz,2H,NCH2),2.43-2.40(m,2H,SCH2CH2),2.34-2.31 (m,2H,CHCH2),2.25-2.17(m,2H,CHCH2),2.09-2.01(m,2H,CCH2),1.98-1.74(m,4H,CCH2CH2).

[0227] 13 C NMR (151MHz, CDCl3) δ (ppm): 170.77, 166.01, 159.45, 157.34, 156.20, 129.05, 105.62, 64.28, 58.80, 51.61, 44.47, 43.88, 31.92, 30.74, 30.08, 18.95, 14.60. Example 6

[0228] Step 1 Synthesis of 1-(tert-butyl)4-(4-nitrophenyl)piperidine-1,4-dicarboxylate (2)

[0229]

[0230] Compound 1 (2.00 g, 8.72 mmol), p-nitrophenol (1.21 g, 8.72 mmol) and acetonitrile (90 mL) were added to a 250 mL single-necked bottle in sequence, stirred to dissolve, and then DCC (1.80 g, 8.72 mmol) was added. After the addition was complete, the reaction was carried out at room temperature under nitrogen protection overnight. TLC (V 石油醚 :V 乙酸乙酯 =3:1) to monitor the complete reaction of the raw materials. Filter the reaction system, concentrate the filtrate under reduced pressure and purify it by rapid preparative chromatography (40 g, V 石油醚 :V 乙酸乙酯 =3:1) to obtain 1.65 g of white solid intermediate 2 with a yield of 54.0%.

[0231] 1H NMR(600MHz,CDCl3)δ(ppm):8.37-8.22(m,2H,ArH),7.32-7.26(m,2H,ArH),4.12(s,2H,NCH2),3.03-2.86(m,2 H,NCH2),2.83-2.71(m,1H,CH),2.06(d,J=13.0Hz,2H,CHCH2),1.85-1.71(m,2H,CHCH2),1.48(s,9H,(CH3)3).

[0232] Step 2 Synthesis of tert-butyl 4-((3,5-dichloropyridin-4-yl)carbamoyl)piperidine-1-carboxylate (3)

[0233]

[0234] To a 50 mL single-mouth bottle, intermediate 2 (1.00 g, 2.85 mmol), 4-amino-3,5-dichloropyridine (930 mg, 5.70 mmol) and N,N-dimethylformamide (15 mL) were added in sequence, and then sodium hydride (60%, dispersed in mineral oil, 205 mg, 5.13 mmol) was added to the reaction system in two batches at 0°C. After the addition was completed, the reaction was carried out at room temperature under nitrogen protection for 4 h. TLC (V 石油醚 :V 乙酸乙酯 =3:1) to monitor the complete reaction of the raw materials. Water (30 mL) was added to the reaction solution to quench, and the mixture was extracted with ethyl acetate (20 mL×3). The organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, concentrated and purified by flash preparative chromatography (20 g, V 石油醚 :V 乙酸乙酯 =1:1) to obtain 950 mg of white solid intermediate 3 with a yield of 89.0%.

[0235] 1 H NMR (600MHz, CDCl3) δ (ppm): 8.53 (d, J = 1.4Hz, 2H, ArH), 4.17 (s, 2H, NCH2), 2.93 -2.79(m,2H,NCH2),2.62-2.51(m,1H,CH),1.98(d,J=13.2Hz,2H,CHCH2),1.87-1.72(m,2H,CHCH2),1.47(s,9H,(CH3)3).

[0236] HRMS(ESI):m / z[M+H] + Theoretical value C 16 H 21Cl2N3O3: 374.1038; Found: 373.0960. Step 3 Synthesis of N-(3,5-dichloropyridin-4-yl)piperidine-4-carboxamide hydrochloride (4)

[0237]

[0238] Add intermediate 3 (950 mg, 2.54 mmol) and dichloromethane (3 mL) to a 50 mL Schlenk reaction tube, stir to dissolve, then add hydrogen chloride 1,4-dioxane solution (4.0 M, 3 mL) dropwise to the reaction system at 0°C, and react at room temperature for 2 h under nitrogen protection. TLC (V 石油醚 :V 乙酸乙酯 =2:1) ​​Monitor the reaction of the starting materials to be complete. Filter the reaction system, wash the filter cake with ethyl acetate (5 mL), and dry to obtain 766 mg of white solid intermediate 4, with a yield of 97.0%.

[0239] 1 H NMR (400 MHz, DMSO-d6) δ (ppm): 10.48 (s, 1H, HCl), 9.24 (s, 1H, NH), 8.85 (s, 1H, NH), 8.69 (s, 2H, ArH), 3.29 (d, J = 13.5 Hz, 2H, NCH2), 3.00-2.89 (m, 2H, NCH2), 2.87-2.77 (m, 1H, CH), 2.09-1.97 (m, 2H, CHCH2), 1.95-1.79 (m, 2H, CHCH2). Step 4 Synthesis of N-(3,5-dichloropyridin-4-yl)-1-(4-((1-(hydroxymethyl)cyclobutyl)amino)-5-oxo-6,7-dihydrothieno[3,2-d]pyrimidin-2-yl)piperidine-4-carboxamide (Example 6)

[0240]

[0241] To 10 mL of Schlenk, intermediate 4 (240 mg, 0.77 mmol), intermediate 8 of Example 1 (202 mg, 0.70 mmol), tetrahydrofuran (3 mL), water (0.75 mL) and N,N-diisopropylethylamine (272 mg, 2.10 mmol) were added in sequence. After the addition was completed, the reaction was carried out at 65° C. under nitrogen protection overnight. TLC (V 二氯甲烷 :V 甲醇 =10:1) Monitor the reaction of the starting materials to be complete. Filter the reaction system, wash the filter cake with tetrahydrofuran (10 mL), and dry to obtain 162 mg of white solid Example 6, with a yield of 44.0%.

[0242] 1H NMR(400MHz,DMSO-d6)δ(ppm):10.22(s,1H,NH),8.67(s,2H,ArH),7.36(s,1H,NH),4.84(t, J=5.7Hz,1H,OH),4.66(d,J=13.0Hz,2H,CH2OH),3.79-3.68(m,2H,SCH2),3.47-3.37(m,1H,H of SCH2CH2),3.26-3.16(m,1H,H of SCH2CH2),3.08-2.98(m,2H,NCH2),2.97-2.77(m,3H,NCH2 and CH),2.41-2.25(m,2H,CHCH2),2.23-2.11(m,2H,CHCH2),1.89(d,J=12.9Hz,2H,CH2CH2CH2),1.83-1.69(m,2H,CH2CH2CH2),1.64-1.49(m,2H,CH2CH2CH2).

[0243] HRMS(ESI):m / z[M+H] + Theoretical value C 22 H 26 Cl2N3O3S: 525.124; Measured value: 525.121. Example 7

[0244] Step 1 Synthesis of tert-butyl 4-(2-hydroxy-5-methylbenzamido)piperidine-1-carboxylate (3)

[0245]

[0246] Compound 1 (1.00 g, 6.57 mmol), compound 2 (1.58 g, 7.88 mmol), HATU (3.00 g, 7.88 mmol), DIPEA (1.70 g, 13.17 mmol) and dichloromethane (20 mL) were added to a 100 mL single-necked bottle in sequence and reacted at room temperature for 1 h under nitrogen protection. TLC (V 石油醚 :V 乙酸乙酯 =2:1) ​​After the reaction was completed, water (20 mL) was added to the reaction solution, and dichloromethane (50 mL×3) was used for extraction. The organic phase was collected. The product was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by flash chromatography (40 g, V 石油醚 :V 乙酸乙酯 =3:1) to obtain 350 mg of white solid intermediate 3 with a yield of 15.9%.

[0247] 1H NMR (400MHz, DMSO-d6) δ (ppm): 12.34 (s, 1H, OH), 8.53 (d, J = 7.8Hz, 1H, ArH), 7.68 (d, J = 2.1Hz, 1H, HN), 7.21 (dd, J1 = 8.4Hz, J2 = 2.1Hz, 1H, ArH), 6 .78(d,J=8.3Hz,1H,ArH),4.07-3.91(m,3H,CH2,CH),2.94-2.74(m,2H,CH2),2.24(s,3H,ArCH3),1.83-1.76(m,2H,CH2),1.41(s,9H,C(CH3)3).

[0248] Step 2 Synthesis of 2-hydroxy-5-methyl-N-(piperidin-4-yl)benzamide trifluoroacetate (4)

[0249]

[0250] In a 25 mL single-necked bottle, intermediate 3 (0.2 g, 0.6 mmol) and dichloromethane (3 mL) were added in sequence, the temperature was lowered to 0°C, TFA (3 mL) was slowly added dropwise, and the reaction was carried out at room temperature for 1 h. TLC (V 石油醚 :V 乙酸乙酯 =2:1) ​​After the reaction was completed, the mixture was concentrated to give 206 mg of yellow oily intermediate 4 with a yield of 99%, which was used directly in the next step.

[0251] Step 3 Synthesis of 2-hydroxy-N-(1-(4-((1-(hydroxymethyl)cyclobutyl)amino)-5-oxo-6,7-dihydrothieno[3,2-d]pyrimidin-2-yl)piperidin-4-yl)-5-methylbenzamide (Example 7)

[0252]

[0253] To a 50 mL Schlenk reaction tube, intermediate 4 (0.10 g, 0.30 mmol), intermediate 8 of Example 1 (0.086 g, 0.3 mmol), DIPEA (0.116 g, 0.90 mmol), THF (4 mL) and H2O (1 mL) were added in sequence, and the mixture was reacted at 65°C overnight under nitrogen protection. TLC (V 二氯甲烷 :V 甲醇 =10:1) after the reaction was completed, the reaction was concentrated and purified by preparative plate (V 二氯甲烷 :V 甲醇 =10:1) to obtain 105 mg of white solid Example 7, with a yield of 71.4%.

[0254] 1H NMR (400MHz, DMSO-d6) δ (ppm): 12.40 (s, 1H, ArOH), 8.51 (d, J = 7.9Hz, 1H, NH), 7.67 (d, J = 2.4Hz, 1H, NH), 7.40 (s, 1H, ArH), 7.20 (dd, J1 = 8.4Hz, J2 = 2.1Hz, 1H,ArH),6.78(d,J=8.3Hz,1H,ArH),4.85(t,J=5.6Hz,1H,OH),4.69-4.56(m ,2H,CH2),4.24-4.12(m,1H,CH),3.78-3.68(m,2H,CH2),3.51-3.39(m,1H,H of CH2),3.26-3.16(m,1H,H of CH2),2.98-2.84(m,2H,CH2),2.41-2.26(m,2H,CH2),2.22(s,3H,CH3),2.20-2.12(m,2H,CH2),1.87-1.71(m,4H,CH2CH2),1.56-1.43(m,2H,CH2).

[0255] 13 C NMR(101MHz,MeOD)δ(ppm):175.89,168.94,162.44,158.13,157.56,134.11,127.97,127.52,1 16.86,115.25,107.24,64.21,58.72,43.14,32.09,31.24,31.20,29.85,29.72,19.14,14.45.

[0256] HRMS(ESI):m / z[M+H] + Theoretical value C 24 H 31 N5O4S: 486.2175; Measured value: 486.2161. Example 8

[0257] Step 1 Synthesis of tert-butyl 6-(5-chloropyrimidin-2-yl)-2,6-diazaspiro[3.3]heptane-2-carboxylate (2)

[0258]

[0259] Compound 1 (700 mg, 3.53 mmol), tert-butyl 2,6-diazaspiro[3.3]heptane-2-carboxylate (848 mg, 3.53 mmol), tetrahydrofuran (20 mL) and triethylamine (429 mg, 4.24 mmol) were added to a 50 mL single-mouth bottle in sequence. After the addition was complete, the reaction was allowed to proceed at room temperature overnight. TLC (V 石油醚:V 乙酸乙酯 =4:1) to monitor the complete reaction of the raw material. The reaction solution was directly concentrated and purified by rapid preparative chromatography (20 g, V 石油醚 :V 乙酸乙酯 =4:1) to obtain 825 mg of white solid intermediate 2 with a yield of 75.2%.

[0260] 1 H NMR(400MHz,DMSO-d6)δ(ppm):8.40(s,2H,ArH),4.16(s,4H,N(CH2)2),4.02(s,4H,N(CH2)2),1.37(s,9H,C(CH3)3).

[0261] Step 2 Synthesis of 2-(5-chloropyrimidin-2-yl)-2,6-diazaspiro[3.3]heptane (3)

[0262]

[0263] To a 50 mL single-necked bottle, intermediate 2 (500 mg, 1.61 mmol), tetrahydrofuran (8 mL) and trifluoroacetic acid (2 mL) were added in sequence, and the reaction was carried out at room temperature for 2 h. TLC (V 二氯甲烷 :V 甲醇 =10:1) monitor the reaction of the raw materials to complete. The reaction solution was concentrated, and the residue was added with saturated sodium bicarbonate aqueous solution (20 mL). After the addition was complete, the mixture was extracted with dichloromethane (20 mL×3), and the organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 320 mg of white solid intermediate 3, which was used directly in the next step without purification.

[0264] Step 3 Synthesis of 2-(6-(5-chloropyrimidin-2-yl)-2,6-diazaspiro[3.3]heptane-2-yl)-4-((1-(hydroxymethyl)cyclobutyl)amino)-6,7-dihydrothieno[3,2-d]pyrimidine 5-oxide (Example 8)

[0265]

[0266] In a 50 mL single-necked bottle, intermediate 8 (228 mg, 0.79 mmol), intermediate 3 (200 mg, 0.95 mmol), N,N-diisopropylethylamine (306 mg, 2.37 mmol) and tetrahydrofuran (10 mL) were added in sequence, and the reaction was carried out at 80° C. under nitrogen protection overnight. TLC (V 二氯甲烷 :V 甲醇 =10:1) to monitor the complete reaction of the raw material. The reaction solution was directly concentrated and purified by rapid preparative chromatography (20 g, V 二氯甲烷 :V 甲醇=10:1) to obtain 210 mg of a white solid (Example 8) with a yield of 44.1%.

[0267] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 8.41 (s, 2H, ArH), 7.36 (s, 1H, NH), 4.85 (t, J = 5.6Hz, 1H, OH), 4.23(s,4H,N(CH2)2),4.19(s,4H,N(CH2)2),3.70(d,J=5.7Hz,2H,SCH2),3.45-3.34(m,1H,H of CH2OH),3.22-3.14(m,1H,H of CH2OH),2.97-2.82(m,2H,SCH2CH2),2.42-2.28(m,2H,H of CH2CH2CH2),2.17-2.06(m,2H,H of CH2CH2CH2),1.85-1.69(m,2H,Hof CH2CH2CH2).

[0268] 13 C NMR(100MHz,DMSO-d6)δ(ppm):175.21,163.44,161.08,158.30,156.47,118.79 ,110.66,64.63,60.60,60.12,59.10,48.97,33.68,32.77,30.32,30.15,14.77.

[0269] HRMS(ESI):m / z[M+H] + Theoretical value C 20 H 24 ClN7O2S: 462.1479; Measured value: 462.1474. Example 9

[0270] Step 1 Synthesis of 2-chloro-N-(3,5-dichloropyridin-4-yl)-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-amine (2)

[0271]

[0272] Compound 1 (500 mg, 2.27 mmol), 4-amino-3,5-dichloropyridine (740 mg, 4.54 mmol), Pd2(dba)3 (211 mg, 0.23 mmol), BINAP (143 mg, 0.23 mmol), potassium tert-butoxide (509 mg, 4.54 mmol) and anhydrous toluene (20 mL) were added to a 100 mL Schlenk reaction tube in sequence, and the mixture was reacted at 100 °C for 15 h under nitrogen protection. TLC (V 石油醚 :V 乙酸乙酯 =3:1) to monitor the reaction completion, and the reaction solution was cooled to room temperature. Filtered, concentrated and subjected to flash preparative chromatography (20 g, V 石油醚 :V 乙酸乙酯 =4:1) to obtain 130 mg of yellow solid intermediate 2 with a yield of 16.5%.

[0273] 1 H NMR (400MHz, CDCl3) δ (ppm): 8.54 (s, 2H, ArH), 6.57 (s, 1H, NH), 3.19-3.15 (m, 2H, SCH2), 2.97-2.92 (m, 2H, CH2CH2CH2), 2.35-2.29 (m, 2H, CH2CH2CH2).

[0274] HRMS(ESI):m / z[M+H] + Theoretical value C 12 H9Cl3N4S: 346.9692; Found: 346.9670. Step 2 Synthesis of 2-chloro-4-((3,5-dichloropyridin-4-yl)amino)-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidine 5,5-dioxide (3)

[0275]

[0276] To a 100 mL single-necked bottle, intermediate 2 (600 mg, 1.73 mmol), m-CPBA (85%, 879 mg, 4.33 mmol) and dichloromethane (20 mL) were added in sequence and stirred at room temperature for 5 h under N2 protection. TLC (V 石油醚 :V 乙酸乙酯 =3:1) monitoring showed that the reaction was complete. Saturated sodium thiosulfate aqueous solution (100 mL) was added, extracted with dichloromethane (100 mL×3), the organic phase was washed with saturated sodium bicarbonate aqueous solution (100 mL×3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated and subjected to flash preparative chromatography (12 g, V 石油醚 :V 乙酸乙酯 =9:1~1:1) to obtain 110 mg of white solid intermediate 3 with a yield of 16.7%.

[0277] 1 H NMR (400MHz, CDCl3) δ (ppm): 8.65 (s, 1H, NH), 8.59 (s, 2H, ArH), 3.54-3.42 (m, 2H, SO2CH2), 3.13-3.04 (m, 2H, CH2CH2CH2), 2.66-2.53 (m, 2H, CH2CH2CH2).

[0278] HRMS(ESI):m / z[MH] - Theoretical value C 12 H9Cl3N4O2S: 376.9434; Found: 376.9456. Step 3 Synthesis of 2-(4-(5-chloropyrimidin-2-yl)piperidin-1-yl)-4-((3,5-dichloropyridin-4-yl)amino)-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidine 5,5-dioxide (Example 9)

[0279]

[0280] To a 100 mL single-necked bottle, intermediate 3 (110 mg, 0.29 mmol), 5-chloro-2-(piperidin-4-yl)pyrimidine (57 mg, 0.29 mmol), tetrahydrofuran (8 mL), water (2 mL) and DIPEA (112 mg, 0.87 mmol) were added in sequence, and the mixture was reacted at 65 °C for 3 h under nitrogen protection. TLC (V 二氯甲烷 :V 甲醇 =50:1) to monitor the completion of the reaction, concentrate and perform flash chromatography (12 g, V 石油醚 :V 乙酸乙酯 =1:1) to obtain 93 mg of white solid Example 9 with a yield of 59.2%.

[0281] 1 H NMR (400MHz, CDCl3) δ (ppm): 8.61 (s, 2H, ArH), 8.58 (s, 1H, NH), 8.51 (s, 2H, ArH), 4.87 (br, 1H, H ofSO2CH2), 4.39 (br, 1H, H of SO2CH2),3.45-3.34(m,2H,NCH2),3.16-3.07(m,1H,CH),3.05-2.83(m,4H,NCH2,CH2CH2 CH2),2.54-2.43(m,2H,CH2CH2CH2),2.03-1.87(m,2H,CHCH2),1.82-1.69(m,2H,CHCH2).

[0282] 13C NMR (101MHz, CDCl3) δ (ppm): 170.43, 166.25, 159.21, 155.62, 155.54, 148.02 ,141.17,129.14,128.73,106.06,51.77,44.64,43.67,31.78,30.66,18.72.

[0283] HRMS(ESI):m / z[M+Na] + Theoretical value C 21 H 20 Cl3N7O2S:562.0363; Measured value:562.0380.

[0284] Embodiment 11

[0285] Step 1 Synthesis of methyl 2-(4-((2-chloro-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)acetate (2)

[0286]

[0287] In a 100 mL single-mouth bottle, add intermediate 8 of Example 5 (150 mg, 0.66 mmol), compound 1 (0.10 mg, 0.55 mmol), Xantphos (48 mg, 0.083 mmol), Na2CO3 (120 mg, 1.10 mmol) and 1,4-dioxane (2 mL) in sequence, replace nitrogen fully, add Pd2(dba)3 (25 mg, 0.028 mmol), continue to replace nitrogen three times after addition. React at 70°C overnight. TLC (V 石油醚 :V 乙酸乙酯 =2:1) ​​After the reaction was completed, ethyl acetate (10 mL) was added to the reaction solution, which was filtered, concentrated and purified by flash preparative chromatography (40 g, V 石油醚 :V 乙酸乙酯 =1:1) to obtain 200 mg of white solid intermediate 2 with a yield of 41.3%.

[0288] 1H NMR (400MHz, DMSO-d6) δ (ppm): 8.71 (s, 1H, NH), 7.46 (dd, J1=12.2Hz, J2=2.1Hz, 1H, ArH), 7.37-7.33 (m, 1H, ArH), 7.29 (t, J=8.4 Hz,1H,ArH),3.70(s,2H,ArCH2),3.63(s,3H,OCH3),3.17-3.12(m,2H,CH2),2.79(t,J=6.4Hz,2H,CH2),2.17-2.10(m,2H,CH2).

[0289] Step 2 Synthesis of methyl 2-(4-(((2-chloro-5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)acetate (3)

[0290]

[0291] To a 25 mL single-necked bottle, intermediate 2 (280 mg, 0.76 mmol) and DCM (10 mL) were added in sequence. At room temperature, m-CPBA (85%, 390 mg, 1.90 mmol) was added in two batches. After the addition was completed, the reaction was carried out at room temperature under nitrogen protection for 2.5 h. TLC (V 石油醚 :V 乙酸乙酯 =1:1) After the reaction was completed, the reaction solution was quenched with saturated sodium thiosulfate solution (10 mL), extracted with DCM (30 mL × 3), and the organic phase was collected. It was washed with saturated sodium bicarbonate solution (30 mL × 3) and saturated brine (30 mL × 3) in sequence, dried over anhydrous sodium sulfate, filtered, concentrated and purified by flash preparative chromatography (20 g, V 石油醚 :V 乙酸乙酯 =1:1) to obtain 202 mg of white solid intermediate 3 with a yield of 72.4%.

[0292] HRMS(ESI):m / z[M+H] + Theoretical value C 16 H 15 ClFN3O4S: 400.0530; theoretical value: 400.0534. Step 3 Synthesis of methyl 2-(4-((2-(4-(5-chloropyrimidin-2-yl)piperidin-1-yl)-5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)acetate (4)

[0293]

[0294] To a 25 mL single-necked bottle, intermediate 3 (210 mg, 0.53 mmol), 5-chloro-2-(piperidin-4-yl)pyrimidine (114 mg, 0.58 mmol), DIPEA (205 g, 1.59 mmol), THF (4 mL) and H2O (1 mL) were added in sequence, and the mixture was reacted at 65°C overnight under nitrogen protection. TLC (V 二氯甲烷 :V 甲醇 =30:1) The reaction was monitored to be complete, concentrated, and slurried with tetrahydrofuran (1 mL) to obtain 130 mg of white solid intermediate 4, with a yield of 87.6%.

[0295] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 8.87 (s, 2H, ArH), 8.68 (s, 1H, NH), 7.61 (dd, J1=12.3Hz, J2=2. 0Hz,1H,ArH),7.31(t,J=8.4Hz,1H,ArH),7.26-7.22(m,1H,ArH),4.80-4.51(m,2H,CH2),3.7 0(s,2H,ArCH2),3.62(s,3H,OCH3),3.59-3.55(m,2H,CH2),3.23-3.12(m,3H,CH2CH2),2.82( t,J=6.4Hz,2H,CH2),2.31-2.26(m,2H,CH2),2.05-2.00(m,2H,CH2),1.75-1.64(m,2H,CH2).

[0296] HRMS(ESI):m / z[M+H] + Theoretical value C 25 H 26 ClFN6O4S:561.1487; Found:561.1479.

[0297] Step 4 Synthesis of 2-(4-((2-(4-(5-chloropyrimidin-2-yl)piperidin-1-yl)-5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)acetic acid (Example 11)

[0298]

[0299] To a 25 mL single-necked bottle, intermediate 4 (150 mg, 0.28 mmol), ethanol (1 mL), tetrahydrofuran (1 mL) and water (0.5 mL) were added in sequence and reacted at room temperature for 2 h under N2 protection. TLC (V 二氯甲烷 :V 甲醇=30:1) monitor the reaction completion, adjust the solution pH to 7 with 1M hydrochloric acid, and concentrate. Add H2O (2mL), adjust the solution pH to 5 with 1M hydrochloric acid, filter, and dry the filter cake to obtain 132 mg of white solid Example 11, with a yield of 90.7%.

[0300] 1 H NMR(400MHz,DMSO-d6)δ(ppm):12.47(s,1H,COOH),8.87(s,2H,ArH),8.67(s,1H,NH),7.5 9(dd,J1=12.2Hz,J2=2.1Hz,1H,ArH),7.30(t,J=8.4Hz,1H,ArH),7.24-7.20(m,1H,ArH), 4.76-4.48(m,2H,CH2),3.59-3.54(m,4H,CH2,CH2),3.24-3.12(m,3H,CH2CH2),2.82(t,J =6.4Hz,2H,CH2),2.32-2.25(m,2H,CH2),2.06-2.00(m,2H,CH2),1.78-1.63(m,2H,CH2).

[0301] 13 C NMR(101MHz,DMSO-d6)δ(ppm):172.33,170.55,167.29,160.75(d,J C-F =243.4Hz),156.23,156.14,138.91(d,J C-F =11.1Hz),132.31(d,J C-F =6.1Hz),129.01,117.78(d,J C-F =17.2Hz),117.17(d,J C-F =3.0Hz),108.48(d,J C-F =27.3Hz),106.03,51.36,44.11,43.91,34.30,31.92,30.63,18.77.

[0302] HRMS(ESI):m / z[M+H] + Theoretical value C 24 H 24 ClFN6O4S:547.1331; Found:547.1300.

[0303] Example 12

[0304] Step 1 Synthesis of 4-azido-3,5-dichloropyridine (2)

[0305]

[0306] Compound 1 (500 mg, 2.7 mmol), DMSO (5 mL) and sodium azide (356 mg, 5.4 mmol) were added to a 25 mL single-necked bottle in sequence, and the mixture was reacted at 30° C. for 36 h under nitrogen protection. TLC (V 石油醚 :V 乙酸乙酯 =5:1) Monitor the reaction of the starting material to be complete, cool, add water (15 mL) to quench the reaction, solid precipitates, filter and dry to obtain 450 mg of bluish white solid intermediate 2, yield 86.9%.

[0307] 1 H NMR (400MHz, CDCl3) δ (ppm): 8.40 (s, 2H, ArH).

[0308] Step 2 Synthesis of 4-((1-(hydroxymethyl)cyclobutyl)amino)-2-((trimethylsilyl)ethynyl)-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidine 5,5-dioxide (3)

[0309]

[0310] Into a 100 mL single-necked bottle, the intermediate 10 of Example 5 (600 mg, 1.89 mmol), acetonitrile (30 mL), trimethylethynylsilane (1.11 g, 11.3 mmol), cuprous iodide (54 mg, 0.28 mmol), triethylamine (0.8 mL, 5.66 mmol) and tetrakis(triphenylphosphine)palladium (327 mg, 0.28 mmol) were added in sequence, and the mixture was reacted at 80° C. for 2 h under nitrogen protection. TLC (V 石油醚 :V 乙酸乙酯 =1:1) monitor the complete reaction of the raw material, cool, directly concentrate, and purify by column chromatography (V 石油醚 :V 乙酸乙酯 =1:1), to obtain 520 mg of yellow solid intermediate 3, with a yield of 72.6%.

[0311] 1H NMR (400 MHz, DMSO-d6) δ (ppm): 6.93 (s, 1H, NH), 5.02 (t, J = 5.6 Hz, 1H, OH), 3.66 (d, J = 5.6 Hz, 2H, CH2OH), 3.63-3.56 (m, 2H, SO2CH2), 2.85 (t, J = 6.4 Hz, 2H, SO2CH2CH2), 2.33-2.23 (m, 4H, SO2CH2CH2CH2, cyclobutane-CH2), 2.17-2.09 (m, 2H, cyclobutane-CH2), 1.92-1.82 (m, 1H, cyclobutane-CH2), 1.80-1.72 (m, 1H, cyclobutane-CH2), 0.24 (s, 9H, (CH3)3).

[0312] Step 3 Synthesis of 2-ethynyl-4-((1-(hydroxymethyl)cyclobutyl)amino)-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidine 5,5-dioxide (4)

[0313]

[0314] To a 100 mL single-necked bottle, add intermediate 3 (520 mg, 1.37 mml), methanol (15 mL) and tetrahydrofuran (15 mL) in sequence, and add lithium hydroxide monohydrate (58 mg, 1.37) in water (15 mL) dropwise under an ice-water bath. After the addition is complete, react at room temperature for 30 min. TLC (V 石油醚 :V 乙酸乙酯 =1:2) monitor the reaction of the raw material, dilute with water (30 mL), extract with ethyl acetate (30 mL×3), combine the organic phases, dry over anhydrous sodium sulfate, filter, concentrate, and purify by column chromatography (V 石油醚 :V 乙酸乙酯 =1:2), to obtain 390 mg of yellow solid intermediate 4, with a yield of 92.6%.

[0315] 1 H NMR (400 MHz, DMSO-d6) δ (ppm): 6.94 (s, 1H, NH), 5.01 (t, J = 5.6 Hz, 1H, OH), 4.31 (s, 1H, CH), 3.67 (d, J = 5.2 Hz, 2H, CH2OH), 3.64-3.56 (m, 2H, SO2CH2), 2.85 (t, J = 6.4 Hz, 2H, SO2CH2CH2), 2.32-2.24 (m, 4H, SO2CH2CH2CH2, cyclobutane-CH2), 2.17-2.09 (m, 2H, cyclobutane-CH2), 1.93-1.82 (m, 1H, cyclobutane-CH2), 1.80-1.73 (m, 1H, cyclobutane-CH2).

[0316] Step 4 Synthesis of 2-(1-(3,5-dichloropyridin-4-yl)-1H-1,2,3-triazol-4-yl)-4-((1-(hydroxymethyl)cyclobutyl)amino)-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidine 5,5-dioxide (Example 12)

[0317]

[0318] Into a 25 mL single-necked bottle, intermediate 2 (231 mg, 1.22 mmol), 4 (250 mg, 0.81 mmol), tert-butyl alcohol (8 mL), water (2 mL), anhydrous copper sulfate (20 mg, 0.08 mmol) and sodium ascorbate (32 mg, 0.16 mmol) were added in sequence, and the mixture was reacted at 60 °C overnight under nitrogen protection. TLC (V 石油醚 :V 乙酸乙酯 =1:2) monitor the reaction of the raw materials, cool to room temperature, dilute with water (20 mL), extract with dichloromethane (15 mL×3), combine the organic phases, dry over anhydrous sodium sulfate, filter, concentrate, and purify by column chromatography (V 二氯甲烷 :V 甲醇 =100:1), to obtain 160 mg of white solid Example 12, with a yield of 39.6%.

[0319] 1 H NMR (400 MHz, DMSO-d6) δ (ppm): 9.19 (s, 1H, ArH), 9.03 (s, 2H, ArH), 7.01 (s, 1H, NH), 5.03 (s, 1H, OH), 3.77 (d, J = 5.6 Hz, 2H, CH2OH), 3.67-3.60 (m, 2H, SO2CH2), 2.98 (t, J = 6.4 Hz, 2H, SO2CH2CH2), 2.46-2.31 (m, 4H, SO2CH2CH2CH2, cyclobutane-CH2), 2.26-2.18 (m, 2H, cyclobutane-CH2), 2.03-1.92 (m, 1H, cyclobutane-CH2), 1.85-1.73 (m, 1H, cyclobutane-CH2).

[0320] 13 C NMR(101MHz,DMSO-d6)δ(ppm):165.66,157.56,156.74,149.58,146.84,139.78, 129.89,129.67,114.37,64.36,59.33,55.38,51.01,31.59,30.21,18.79,14.45.

[0321] HRMS(ESI):m / z[M+H]+ Theoretical value C 19 H 19 Cl2N7O3S: 496.0725; Measured value 496.0711. Example 13

[0322] Step 1 Synthesis of tert-butyl 5-(((trifluoromethyl)sulfonyl)oxy)-2-azabicyclo[2.2.2]oct-5-ene-2-carboxylate (2)

[0323]

[0324] Compound 1 (1.30 g, 5.77 mmol) and tetrahydrofuran (40 mL) were added to a 50 mL single-mouth bottle in sequence. Under nitrogen protection, LiHMDS (11.5 mL, 11.5 mmol, 1 M in THF) was added dropwise at -78 °C, and the temperature was kept for 30 min. Then, a solution of 2-[N, n-bis(trifluoromethanesulfonyl)amino]-5-chloropyridine (3.40 g, 8.66 mmol) in tetrahydrofuran (10 mL) was added dropwise. After the addition was completed, the temperature was slowly raised to room temperature and the reaction was allowed to proceed overnight. TLC (V 石油醚 :V 乙酸乙酯 =5:1) monitor the reaction of the raw material, add water (30 mL) to quench the reaction, extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated sodium chloride solution (50 mL), dry with anhydrous sodium sulfate, filter, concentrate, and purify with column chromatography (V 石油醚 :V 乙酸乙酯 =20:1), to obtain 1.96 g of yellow oily intermediate 2, with a yield of 95.0%.

[0325] Step 2 Synthesis of tert-butyl 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-azabicyclo[2.2.2]oct-5-ene-2-carboxylate (3)

[0326]

[0327] In a 100 mL single-necked bottle, intermediate 2 (2.30 g, 6.44 mmol), 1,4-dioxane (45 mL), biboric acid pinacol ester (1.96 g, 7.72 mmol), potassium acetate (1.90 g, 19.3 mmol) and Pd(dppf)Cl2 (470 mg, 0.64 mmol) were added in sequence, and the mixture was reacted at 60°C for 3 h under nitrogen protection. TLC (V 石油醚 :V 乙酸乙酯=5:1) monitor the reaction completion of the raw material, cool, add water (30 mL) to quench the reaction, extract with ethyl acetate (30 mL×3), combine the organic phases, wash with saturated sodium chloride solution (50 mL), dry over anhydrous sodium sulfate, filter and concentrate to obtain 2.70 g of black oily intermediate 3, which was directly used in the next step with 80% purity.

[0328] Step 3 Synthesis of tert-butyl 5-(5-chloropyrimidin-2-yl)-2-azabicyclo[2.2.2]oct-5-ene-2-carboxylate (4)

[0329]

[0330] Into a 50 mL single-necked bottle, intermediate 3 (2.70 g, 6.44 mmol, 80% wt), 1,4-dioxane / water (40 mL / 5 mL), 5-chloro-2-iodopyrimidine (1.86 g, 7.72 mmol), sodium carbonate (2.05 g, 19.3 mmol) and tetrakis(triphenylphosphine)palladium (675 mg, 0.64 mmol) were added in sequence, and the mixture was reacted at 90°C overnight under nitrogen protection. TLC (V 石油醚 :V 乙酸乙酯 =5:1) monitor the reaction of the raw material to be complete, cool, add water (30 mL) to quench the reaction, extract with ethyl acetate (30 mL×3), combine the organic phases, wash with saturated sodium chloride solution (50 mL), dry with anhydrous sodium sulfate, filter, concentrate, and purify with column chromatography (V 石油醚 :V 乙酸乙酯 =10:1), to obtain 1.40 g of yellow oily intermediate 4, with a two-step yield of 67.7%.

[0331] 1 H NMR (400MHz, CDCl3) δ (ppm): 8.64 (s, 2H, ArH), 7.60-7.52 (m, 1H, ArH), 5.00-4.78 (br, 1H, NCH), 3.88-3.80 (m, 1H, NCH2CH), 3.39-3.34 (m, 1H, H of NCH2),3.12-3.02(m,1H,H of CH2CH2),2.10-2.04(m,1H,H of CH2CH2),1.82-1.74(m,1H,H of CH2CH2),1.48-1.46(m,2H,H of CH2CH2),1.45-1.40(m,9H,(CH3)3).

[0332] Step 4 Synthesis of tert-butyl 5-(5-chloropyrimidin-2-yl)-2-azabicyclo[2.2.2]octane-2-carboxylate (5)

[0333]

[0334] Intermediate 4 (0.90 g, 2.80 mmol), ethyl acetate (30 mL) and platinum dioxide (90 mg) were added to a 50 mL single-necked bottle in sequence, and the mixture was reacted at room temperature overnight under a hydrogen atmosphere (15 psi). TLC (V 石油醚 :V 乙酸乙酯 =5:1) monitor the complete reaction of the raw materials, cool, filter through diatomaceous earth, concentrate, and purify by column chromatography (V 石油醚 :V 乙酸乙酯 =15:1), to obtain 390 g of white solid intermediate 5, with a yield of 43.0%.

[0335] 1 H NMR (400MHz, CDCl3) δ (ppm): 8.66-8.58 (m, 2H, ArH), 4.19 (s, 0.5H, NCH), 4.06 (s, 0.5H, NCH), 3.38-3.23(m,2H,NCH2),3.24-3.15(m,1H,NCH2CH),2.64-2.56(m,1H,CH),2.41-2.32(m,1H,H of CH2CH2),2.14-1.90(m,2H,CH2),1.87-1.66(m,3H,H of CH2CH2),1.45(d,J=11.2Hz,9H,(CH3)3).

[0336] Step 5 Synthesis of 5-(5-chloropyrimidin-2-yl)-2-azabicyclo[2.2.2]octane hydrochloride (6)

[0337]

[0338] To a 50 mL single-necked bottle, intermediate 5 (200 mg, 0.62 mmol) and 4N HCl / EA (5 mL) were added in sequence, and the mixture was reacted at room temperature for 1 h under nitrogen protection. TLC (V 石油醚 :V 乙酸乙酯 =5:1) The reaction of the starting material was monitored to be complete and the product was directly concentrated to obtain 137 g of white solid intermediate 6, which was used directly in the next step without purification.

[0339] Step 6 Synthesis of 2-(5-(5-chloropyrimidin-2-yl)-2-azabicyclo[2.2.2]octan-2-yl)-4-((1-(hydroxymethyl)cyclobutyl)amino)-6,7-dihydrothieno[3,2-d]pyrimidine 5-oxide (Example 13)

[0340]

[0341] To a 50 mL single-necked bottle, intermediate 6 (137 mg, 0.61 mmol), intermediate 8 (177 mg, 0.61 mmol) of Example 1, tetrahydrofuran / water (6 mL / 1.5 mL) and DIPEA (238 mg, 1.84 mmol) were added in sequence, and the mixture was reacted at 65° C. overnight under nitrogen protection. TLC (V 二氯甲烷 :V 甲醇 =10:1) monitor the reaction of the raw materials, cool, add water (20 mL), extract with ethyl acetate (30 mL×3), combine the organic phases, wash with saturated sodium chloride solution (50 mL), dry with anhydrous sodium sulfate, filter, concentrate, and purify by column chromatography (V 二氯甲烷 :V 甲醇 =50:1) and then purified by preparation plate (V 二氯甲烷 :V 甲醇 =10:1) to obtain 175 mg of white solid Example 13, with a two-step yield of 59.7%.

[0342] 1 H NMR(400MHz,DMSO-d6)δ(ppm):8.95-8.80(m,2H,ArH),7.19(s,1H,NH),4.84-4.82(m,1H, OH),4.79-4.69(m,1H,NCH),3.72-3.71(m,1H,NCH2CH),3.66-3.61(m,1H,CH),3.42-3.32 (m, 4H, CH2OH, NCH2CH), 3.29-3.09 (m, 2H, SOCH2CH2), 2.94-2.79 (m, 2H, SOCH2CH2), 2.41-2.22 (m, 3H, azabicyclo-octane-CH2), 2.17-2.00 (m, 3H, azabicyclo-octane-CH2), 1.90-1.63 (m, 6H, cyclobutane-CH2).

[0343] 13 C NMR(101MHz,DMSO-d6)δ(ppm):175.42,175.00,170.35,161.48,158.17,157.81,155.92,128.80,109.5 5,109.09,64.29,58.90,48.92,46.18,43.77,43.03,42.52,32.78,31.94,31.03,30.13,25.59,14.85.

[0344] HRMS(ESI):m / z[M+H] + Theoretical value C 22 H 27ClN6O2S: 475.1683; Measured value 475.1696 Example 14

[0345] Step 1 Synthesis of methyl 2-(4-((2-chloro-6,7-dihydrothieno[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)acetate (3)

[0346]

[0347] Compound 1 (6.90 g, 33.3 mmol), 2 (6.10 g, 33.3 mmol), 1,4-dioxane (130 mL), Pd2(dba)3 (1.14 g, 1.25 mmol), Xantphos (1.45 g, 2.50 mmol) and sodium carbonate (7.06 g, 66.6 mmol) were added to a 500 mL single-necked bottle in sequence, and the mixture was reacted at 75° C. overnight under nitrogen protection. TLC (V 石油醚 :V 乙酸乙酯 =3:1) monitor the complete reaction of the raw material, cool, filter, concentrate, and purify by column chromatography (V 石油醚 :V 乙酸乙酯 =3:1), to obtain 6.50 g of white solid intermediate 3, with a yield of 55.1%.

[0348] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 9.29 (s, 1H, NH), 7.52 (dd, J1 = 12.4Hz, J2 = 2.0Hz, 1H, ArH), 7.36 (dd, J1 = 8.4Hz, J2 = 2.0Hz, 1H, ArH), 7.29(t,J=8.4Hz,1H,ArH),3.69(s,2H,CH2COOCH3),3.63(s,3H,COOCH3),3.46-3.39(m,2H,SCH2),3.25(t,J=8.4Hz,2H,SCH2CH2).

[0349] Step 2 Synthesis of methyl 2-(4-((2-chloro-5-oxo-6,7-dihydrothieno[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)acetate (4)

[0350]

[0351] Into a 50 mL single-mouth bottle, intermediate 3 (2.00 g, 5.6 mmol), dichloromethane (15 mL), S-(-)-BINOL (160 mg, 0.56 mmol), tetraisopropyl titanate (80 mg, 0.28 mmol) and water (100 mg, 5.6 mmol) were added in sequence, and the mixture was reacted at room temperature for 1 h under nitrogen protection. Then, 70% wt tert-butyl peroxide (800 mg, 6.1 mmol) was added dropwise. After the addition was completed, the reaction was continued for 2 h. TLC (V 二氯甲烷 :V 甲醇 =20:1) monitor the complete reaction of the raw material, wet sample loading, and column chromatography purification (V 二氯甲烷 :V 甲醇 =100:1), to obtain 1.85 g of yellow solid intermediate 4, with a yield of 88.4%.

[0352] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 10.56 (s, 1H, NH), 7.55 (dd, J1=12.0Hz, J2=2.0Hz, 1H, ArH), 7.44-7.33 (m, 2H, ArH), 3.73 (s, 2H, CH2COOCH3), 3.72-3.65 (m, 1H, H ofSOCH2),3.64(s,3H,COOCH3),3.49-3.42(m,1H,H of SOCH2),3.30-3.23(m,1H,H ofSOCH2CH2),3.17-3.11(m,1H,H ofSOCH2CH2).

[0353] Step 3 Synthesis of methyl 2-(4-((2-(4-(5-chloropyrimidin-2-yl)-3,6-dihydropyridin-1(2H)-yl)-5-oxo-6,7-dihydrothieno[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)acetate (5)

[0354]

[0355] To a 250 mL single-necked bottle, intermediate 4 (2.00 g, 5.4 mmol), intermediate 3 (1.32 g, 5.7 mmol) of Example 1, tetrahydrofuran / water (40 mL / 10 mL) and DIPEA (2.9 mL, 16.3 mmol) were added in sequence, and the mixture was reacted at 65° C. overnight under nitrogen protection. TLC (V 二氯甲烷 :V 甲醇 =15:1) monitor the complete reaction of the raw material, cool, concentrate, and purify by column chromatography (V 二氯甲烷 :V 甲醇 =100:1), and then beaten twice (30mL×2, V 石油醚 :V乙酸乙酯 =1:1, V 乙酸乙酯 :V 二氯甲烷 =5:1) to obtain 930 mg of white solid intermediate 5 with a yield of 32.5%.

[0356] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 9.72 (s, 1H, NH), 8.90 (s, 2H, ArH), 7.67 (d, J = 10.4Hz, 1H, ArH), 7.56 (d, J = 8.4Hz, 1H, ArH), 7.35-7.2 8(m,2H,ArH and C=CH),4.50(br,2H,NCH2CH),4.01(br,2H,NCH2CH2),3.71(s,2H,CH2COOCH3),3.64(s,3H,COOCH3),3.59-3.51(m,1H,H of SOCH2),3.30-3.26(m,1H,H of SOCH2),3.12-3.06(m,1H,H of SOCH2CH2),3.03-2.96(m,1H,H of SOCH2CH2),2.70(br,2H,NCH2CH2).

[0357] Step 4 Synthesis of 2-(4-((2-(4-(5-chloropyrimidin-2-yl)-3,6-dihydropyridin-1(2H)-yl)-5-oxo-6,7-dihydrothieno[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)acetic acid (Example 14)

[0358]

[0359] To a 25 mL single-necked bottle, intermediate 5 (450 mg, 0.85 mmol), acetic acid (4.5 mL) and 37% concentrated hydrochloric acid (1.5 mL) were added in sequence, and the mixture was reacted at 60°C for 4 h under nitrogen protection. TLC (V 二氯甲烷 :V 甲醇 =20:1) monitor the complete reaction of the raw materials, cool, directly concentrate, add water (20mL), solid precipitate, filter, and then slurry (15mL, V 二氯甲烷 :V 甲醇 =50:1) purification to give 250mg of white solid Example 14, yield 57.0%.

[0360] 1H NMR(400MHz,DMSO-d6)δ(ppm):12.43(s,1H,COOH),9.71(s,1H,NH),8.90(s,2H,A rH),7.65(d,J=10.4Hz,1H,ArH),7.54(d,J=8.4Hz,1H,ArH),7.33-7.28(m,2H,ArH andC=CH),4.50(br,2H,NCH2CH),4.02(br,2H,NCH2CH2),3.59(s,2H,CH2COOH),3.59-3.51(m,1H,H of SOCH2),3.30-3.26(m,1H,H ofSOCH2),3.13-3.06(m,1H,H of SOCH2) SOCH2CH2),3.02-2.96(m,1H,H of SOCH2CH2),2.70(br,2H,NCH2CH2).

[0361] Embodiment 15

[0362] Step 1 Synthesis of methyl 2-(4-((2-(4-(5-chloropyrimidin-2-yl)piperazin-1-yl)-5-oxo-6,7-dihydrothieno[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)acetate (2)

[0363]

[0364] Into a 25 mL single-necked bottle, intermediate 4 of Example 14 (307 mg, 0.83 mmol), THF (8 mL), H2O (2 mL), 5-chloro-2-(piperazin-1-yl)pyrimidine (1,180 mg, 0.90 mmol) and DIPEA (322 mg, 2.49 mmol) were added in sequence. After the addition was completed, nitrogen was protected and the mixture was kept at 65°C overnight. TLC (V 二氯甲烷 :V 甲醇 =15:1) to monitor the complete reaction of the raw materials, the reaction solution was concentrated and subjected to rapid preparative chromatography (12 g, V 二氯甲烷 :V 甲醇 =50:1) to obtain 395 mg of white solid intermediate 2 with a yield of 90.2%.

[0365] 1H NMR (400MHz, DMSO-d6) δ (ppm): 9.73 (s, 1H, NH), 8.44 (s, 2H, ArH), 7.59 (d, J = 12.8Hz, 1H, ArH), 7.51 (d, J = 8.4Hz, 1H, ArH), 7 .30(t,J=8.8Hz,1H,ArH),3.91-3.76(m,8H,N(CH2CH2)2),3.69(s,2H,CH2COOCH3),3.63(s,3H,COOCH3),3.57-3.48(m,1H,H ofSOCH2),3.27-3.25(m,1H,H of SOCH2),3.11-3.03(m,1H,Hof SOCH2CH2),3.01-2.95(m,1H,H of SOCH2CH2).

[0366] Step 2 Synthesis of 2-(4-((2-(4-(5-chloropyrimidin-2-yl)piperazin-1-yl)-5-oxo-6,7-dihydrothieno[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)acetic acid (Example 15)

[0367]

[0368] To a 50 mL single-necked bottle, intermediate 2 (300 mg, 0.58 mmol), MeOH (3.7 mL), THF (3.7 mL), H2O (2.7 mL) and NaOH (108 mg, 2.70 mmol) were added in sequence, and the mixture was reacted at 35°C for 4 h under nitrogen protection. TLC (V 二氯甲烷 :V 甲醇 =10:1) Monitor the reaction of the starting material, add water (10 mL) to quench, adjust the pH to 5 with dilute HCl (1 M), and a light yellow solid precipitates. Filter, slurry the solid with MeOH (10 mL) for 4 h, and filter to obtain 200 mg of light yellow solid Example 15, with a yield of 68.41%.

[0369] 1H NMR (400MHz, DMSO-d6) δ (ppm): 12.42 (s, 1H, COOH), 9.73 (s, 1H, NH), 8.46 (s, 2H, ArH), 7.57 (d, J=12.8Hz, 1H, ArH), 7.50 (d ,J=6.4Hz,1H,ArH),7.29(t,J=8.6Hz,1H,ArH),3.95-3.77(m,8H,N(CH2CH2)2),3.59(s,2H,CH2COOH),3.57-3.50(m,1H,H of SOCH2),3.28-3.26(m,1H,H of SOCH2),3.11-3.05(m,1H,H of SOCH2CH2),3.02-2.97(m,1H,H ofSOCH2CH2).

[0370] Example 16

[0371] Step 1 Synthesis of 2-(4-((2-(4-(5-chloropyrimidin-2-yl)piperazin-1-yl)-5-oxo-6,7-dihydrothieno[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)acetic acid sodium salt (Example 16)

[0372]

[0373] To a 25 mL single-necked bottle, add Example 15 (150 mg, 0.29 mmol), water (2.5 mL) and sodium hydroxide (13 mg, 0.32 mmol) in sequence, under nitrogen protection, react at room temperature for 0.5 h, the solid slowly dissolved, cool in an ice-water bath, add acetone (15 mL) after 30 min, a white solid precipitates, continue stirring for 30 min, then filter and dry to obtain 105 mg of white solid Example 16, with a yield of 67.2%.

[0374] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 9.68 (s, 1H, NH), 8.45 (s, 2H, ArH), 7.43 (d, J = 12.4Hz, 1H, ArH), 7.36 ( d,J=8.4Hz,1H,ArH),7.20(t,J=8.4Hz,1H,ArH),3.91-3.78(m,8H,N(CH2CH2)2),3.57-3.49(m,1H,H of SOCH2),3.30-3.24(m,1H,Hof SOCH2),3.18(s,2H,CH2COONa),3.09-3.03(m,1H,H of SOCH2CH2),3.00-2.95(m,1H,H of SOCH2CH2).

[0375] Embodiment 17

[0376] Step 1 Synthesis of methyl 2-(4-((2-(4-(5-chloropyrimidin-2-yl)piperidin-1-yl)-5-oxo-6,7-dihydrothieno[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)acetate (2)

[0377]

[0378] Into a 25 mL single-necked bottle, intermediate 4 of Example 14 (307 mg, 0.83 mmol), THF (8 mL), H2O (2 mL), 5-chloro-2-(piperidin-4-yl)pyrimidine (1,180 mg, 0.91 mmol) and DIPEA (322 mg, 2.49 mmol) were added in sequence. After the addition was completed, nitrogen was protected and the mixture was kept at 65°C overnight. TLC (V 二氯甲烷 :V 甲醇 =15:1) to monitor the complete reaction of the raw materials, the reaction solution was concentrated and subjected to rapid preparative chromatography (12 g, V 二氯甲烷 :V 甲醇 =50:1) to obtain 320 mg of yellow solid intermediate 2 with a yield of 73.85%.

[0379] Step 2 Synthesis of 2-(4-((2-(4-(5-chloropyrimidin-2-yl)piperidin-1-yl)-5-oxo-6,7-dihydrothieno[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)acetic acid (Example 17)

[0380]

[0381] To a 25 mL single-necked bottle, intermediate 2 (200 mg, 0.39 mmol), MeOH (2.5 mL), THF (2.5 mL), H2O (1.8 mL) and NaOH (71 mg, 1.78 mmol) were added in sequence. After the addition was complete, nitrogen was protected and the reaction was carried out at 35°C for 4 h. TLC (V 二氯甲烷 :V 甲醇 =10:1) Monitor the reaction of the raw material, add water (10 mL) to quench the reaction, adjust the pH to 5 with dilute HCl (1N), and a light yellow solid precipitates. After filtration, slurry with MeOH (10 mL) for 4 h, filter and dry to obtain 115 mg of light yellow solid Example 17, with a yield of 60.52%.

[0382] 1H NMR (400MHz, DMSO-d6) δ (ppm): 12.40 (s, 1H, COOH), 9.68 (s, 1H, NH), 8.87 (s, 2H, ArH), 7.62 (d, J = 12.8Hz, 1H, ArH), 7.47 (d, J = 8.4Hz, 1H, ArH), 7.26(t,J=8.4Hz,1H,ArH),4.69(br,2H,SOCH2),3.57(s,2H,CH2COOH),3.55-3.48(m,1H,CH),3.29-3.13(m,4H,CH2NCH2),3.09-3.02(m,1H,H of SOCH2CH2),3.01-2.95(m,1H,H of SOCH2CH2),2.07-1.98(m,2H,NCH2CH2),1.78-1.64(m,2H,NCH2CH2).

[0383] HRMS(ESI):m / z[M+H] + Theoretical value C 22 H 24 ClFN6O3S: 517.1225; Measured value: 517.1194. Example 18

[0384] Step 1 Synthesis of methyl 2-(4-((2-(4-(5-chloropyrimidin-2-yl)-3,6-dihydropyridin-1(2H)-yl)-5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)acetate (2)

[0385]

[0386] Into a 250 mL single-necked bottle, compound Example 11 intermediate 3 (2.00 g, 5.0 mmol), Example 1 intermediate 3 (1.28 g, 5.5 mmol), tetrahydrofuran / water (40 mL / 4 mL) and DIPEA (2.7 mL, 15.0 mmol) were added in sequence, and the mixture was reacted at 65° C. overnight under nitrogen protection. TLC (V 二氯甲烷 :V 甲醇 =20:1) monitor the reaction of the raw materials, cool, add water (20 mL), extract with ethyl acetate (30 mL×3), combine the organic phases, wash with saturated sodium chloride solution (50 mL), dry with anhydrous sodium sulfate, filter, concentrate, and purify by column chromatography (V 二氯甲烷 :V 甲醇 =100:1) and then slurried (150 mL, V 石油醚 :V 乙酸乙酯 =2:1) ​​purification to obtain 850 mg of white solid intermediate 2 with a yield of 30.4%.

[0387] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 8.89 (s, 2H, ArH), 8.70 (s, 1H, NH), 7.63 (d, J = 12 .0Hz,1H,CH),7.38-7.25(m,3H,ArH),4.46(br,2H,NCH2),3.96(br,2H,NCH2), 3.72(s,2H,COCH2),3.64(s,3H,OCH3),3.58-3.55(m,2H,SO2CH2),2.84(t,J=6 .4Hz,2H,SO2CH2CH2),2.68(s,2H,SO2CH2CH2CH2),2.33-2.25(m,2H,NCH2CH2).

[0388] Step 2 Synthesis of 2-(4-((2-(4-(5-chloropyrimidin-2-yl)-3,6-dihydropyridin-1(2H)-yl)-5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)acetic acid (Example 18)

[0389]

[0390] To a 25 mL single-necked bottle, intermediate 2 (50 mg, 0.09 mmol), acetic acid (2.0 mL) and 37% concentrated hydrochloric acid (1.0 mL) were added in sequence, and the mixture was reacted at 80°C for 1 h under nitrogen protection. TLC (V 二氯甲烷 :V 甲醇 =20:1) Monitor the reaction of the starting material to be complete, cool, directly concentrate, add water (10 mL), solid precipitates, filter and dry to obtain 35 mg of white solid Example 18, yield 71.8%.

[0391] 1 H NMR(400MHz,DMSO-d6)δ(ppm):12.44(s,1H,COOH),8.89(s,2H,ArH),8.70(s,1H,NH),7.62(d,J=12.0Hz,1H,CH),7.35-7.24(m,3H,ArH),4.46(br,2H,NC H2),3.97(br,2H,NCH2),3.64-3.54(m,4H,SO2CH2),2.84(t,J=6.4Hz,2H,SO2CH2CH2),2.68(d,J=6.4Hz,2H,SO2CH2CH2CH2),2.33-2.25(m,2H,NCH2CH2).

[0392] 13 C NMR (101MHz, DMSO-d6) δ (ppm): 172.24, 167.11, 160.70 (d, J = 240.5Hz), 156.09, 155.89, 138.91, 132.29, 128 .83,117.71(d,J=13.4Hz),117.27,108.55(d,J=27.0Hz),106.78,51.29,44.61,34.20,31.85,25.18,18.71.

[0393] HRMS(ESI):m / z[M+H] + Theoretical value C 24 H 22 ClFN6O4S:545.1174; Found: 545.1160.

[0394] Embodiment 19

[0395] Step 1 Synthesis of 2-(4-((2-(4-(5-chloropyrimidin-2-yl)-3,6-dihydropyridin-1(2H)-yl)-5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)ammonium acetate (Example 19)

[0396]

[0397] Example 18 (80 mg, 0.15 mmol) and water (5 mL) were added to a 25 mL single-necked bottle in sequence. The solid did not dissolve. 27% ammonia water was added dropwise under an ice-water bath to adjust the pH to 8. After stirring for 15 min, the system became clear. Acetone (10 mL) was added and stirring was continued. A white solid slowly precipitated. After 30 min, it was filtered and dried to obtain 40 mg of white solid Example 19 with a yield of 48.5%.

[0398] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 8.89 (s, 2H, ArH), 7.58 (d, J = 12.1Hz, 1H, CH), 7.35-7.22 (m, 3H, ArH), 4.46 (br, 2H, NCH2), 3.96 (br, 2H, NCH2), 3 .61-3.53(m,2H,COCH2),3.49(s,2H,SO2CH2),2.84(t,J=6.4Hz,2H,SO2CH2CH2),2.68(s,2H,SO2CH2CH2CH2),2.29(t,J=6.0Hz,2H,NCH2CH2).

[0399] Embodiment 20

[0400] Step 1 Synthesis of 2-(4-((2-(4-(5-chloropyrimidin-2-yl)piperidin-1-yl)-5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)ammonium acetate (Example 20)

[0401]

[0402] Example 11 (300 mg, 0.55 mmol), THF (4.5 mL) and methanol (4.5 mL) were added to a 50 mL single-mouth bottle in sequence and stirred at room temperature until homogeneous. Ammonia water (0.1 mL) was slowly dripped into the above reaction system and reacted overnight at room temperature. The system gradually became turbid from clear, filtered, and the filter cake was rinsed with water (2 mL × 3) and collected. 200 mg of white solid Example 20 was obtained by air drying at 50 ° C overnight, with a yield of 64.5%.

[0403] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 8.86 (s, 2H, ArH), 8.63 (s, 1H, NH), 7.56 (dd, J1=12.2Hz, J2=2.1Hz, 1H ,ArH),7.27(t,J=8.4Hz,1H,ArH),7.19(dd,J1=8.3Hz,J2=2.1Hz,1H,ArH),4.60(br,2H,SO2CH2),3. 58-3.54(m,2H,NCH2),3.52(s,2H,ArCH2CO),3.23-3.11(m,3H,NCH2 and CH),2.81(t,J=6.4Hz,2H,SO2C H2CH2CH2),2.31-2.24(m,2H,SO2CH2CH2),2.04-2.00(m,2H,NCH2CH2),1.70-1.67(m,2H,NCH2CH2).

[0404] HRMS(ESI):m / z[M+H] + Theoretical value C 24 H 24 ClFN6O4S: 547.1331; Measured value: 547.1342. Example 21

[0405] Step 1 Synthesis of dimethyl 2-(2-chloro-4-nitrophenyl)malonate (2)

[0406]

[0407] Compound 1 (5.00 g, 28.51 mmol), NMP (100 mL), dimethyl malonate (4.78 g, 36.22 mmol) and sodium hydroxide (2.40 g, 59.85 mmol) were added to a 250 mL reaction bottle in sequence, and reacted at 80° C. for 2 h under nitrogen protection. TLC (V 石油醚 :V 乙酸乙酯 =10:1) monitor the reaction to completion. The reaction solution was cooled to room temperature, quenched with water (300 mL), and adjusted to pH 3 with dilute hydrochloric acid (1N) under ice bath conditions. Solids precipitated and were filtered. The filter cake was rinsed with water (50 mL). The filter cake was collected and dried in vacuo for 2 h to obtain 7.90 g of yellow solid intermediate 2, with a yield of 85.4%.

[0408] 1 H NMR (400MHz, CDCl3) δ (ppm): 8.30 (d, J = 2.4Hz, 1H, ArH), 8.15 (dd, J1 = 8.6Hz, J2 = 2.4Hz, 1H, ArH), 7.74 (d, J = 8.6Hz, 1H, ArH), 5.31 (s, 1H, CH), 3.81 (s, 6H, 2CH3).

[0409] Step 2 Synthesis of methyl 2-(2-chloro-4-nitrophenyl)acetate (3)

[0410]

[0411] To a 50 mL reaction bottle, intermediate 2 (1.00 g, 3.48 mmol), DMSO (10 mL), water (0.06 mL) and sodium chloride (366 mg, 6.26 mmol) were added in sequence, and the mixture was reacted at 110° C. for 16 h under nitrogen protection. TLC (V 石油醚 :V 乙酸乙酯 =5:1) to monitor the reaction completion. The reaction solution was cooled to room temperature, quenched with water (30 mL), extracted with ethyl acetate (50 mL×3), the organic phases were combined, washed with saturated brine (50 mL×4), the organic phases were collected, dried over anhydrous sodium sulfate, filtered, concentrated and purified by flash preparative chromatography (40 g, V 石油醚 :V 乙酸乙酯 =5:1), to obtain 320 mg of yellow oily intermediate 3, with a yield of 40.1%.

[0412] 1H NMR (400MHz, CDCl3) δ (ppm): 8.28 (d, J = 2.4Hz, 1H, ArH), 8.11 (dd, J1 = 8.4Hz, J2 = 2.4Hz, 1H, ArH), 7.50 (d, J = 8.4Hz, 1H, ArH), 3.88 (s, 2H, CH2), 3.74 (s, 3H, CH3).

[0413] Step 3 Synthesis of methyl 2-(4-amino-2-chlorophenyl)acetate (4)

[0414]

[0415] To a 25 mL reaction bottle, intermediate 3 (320 mg, 1.39 mmol), ethanol (4 mL), water (1 mL), iron powder (389 mg, 6.97 mmol) and ammonium chloride (38 mg, 0.70 mmol) were added in sequence and reacted at 85 °C for 1 h under nitrogen protection. TLC (V 石油醚 :V 乙酸乙酯 =3:1) to monitor the complete reaction of the raw materials. The reaction solution was filtered through diatomaceous earth, the filter cake was rinsed with ethyl acetate (50 mL), the filtrate was collected, concentrated and purified by rapid preparative chromatography (12 g, V 石油醚 :V 乙酸乙酯 =3:1), to obtain 220 mg of yellow oily intermediate 4, with a yield of 75.5%.

[0416] Step 4 Synthesis of methyl 2-(2-chloro-4-((2-chloro-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)phenyl)acetate (5)

[0417]

[0418] To a 25 mL single-necked bottle, intermediate 4 (210 mg, 1.05 mmol), 1,4-dioxane (5 mL), intermediate 8 of Example 5 (280 mg, 1.26 mmol), sodium carbonate (223 mg, 2.10 mmol), Xantphos (91 mg, 0.16 mmol) and Pd2(dba)3 (72 mg, 0.08 mmol) were added in sequence, and the mixture was reacted at 70°C for 16 h under nitrogen protection. TLC (V 乙酸乙酯 :V 石油醚 =3:1) to monitor the complete reaction of the raw materials. The reaction solution was filtered through diatomaceous earth, the filter cake was rinsed with ethyl acetate (50 mL), the filtrate was collected, concentrated and purified by rapid preparative chromatography (12 g, V 石油醚 :V 乙酸乙酯 =3:1), to obtain 290 mg of white solid intermediate 5, with a yield of 57.6%.

[0419] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 8.70 (s, 1H, NH), 7.69 (d, J = 2.2Hz, 1H, ArH), 7.50 (dd, J1 = 8.4Hz, J2 = 2.2Hz, 1H, ArH), 7.36 (d, J = 8.4Hz, 1H, ArH) H),3.79(s,2H,CH2COOCH3),3.64(s,3H,COOCH3),3.16-3.09(m,2H,SCH2),2.79(t,J=6.3Hz,2H,SCH2CH2CH2),2.16-2.10(m,2H,SCH2CH2).

[0420] Step 5 Synthesis of methyl 2-(2-chloro-4-((2-chloro-5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)phenyl)acetate (6)

[0421]

[0422] To a 50 mL single-necked bottle, intermediate 5 (280 mg, 0.73 mmol), DCM (10 mL) and m-chloroperbenzoic acid (85%, 371 mg, 3.84 mmol) were added in sequence and reacted at room temperature for 2 h under nitrogen protection. TLC (V 乙酸乙酯 :V 石油醚 =2:1) ​​to monitor the complete reaction of the raw materials. Add saturated sodium thiosulfate (10 mL) to quench, extract with dichloromethane (30 mL×3), combine the organic phases, wash with saturated brine (50 mL×3), collect the organic phases, dry, filter, concentrate and purify by flash preparative chromatography (12 g, V 二氯甲烷 :V 甲醇 =100:1), to obtain 230 mg of white solid intermediate 6, with a yield of 75.9%.

[0423] Step 6 Synthesis of methyl 2-(2-chloro-4-((2-(4-(5-chloropyrimidin-2-yl)piperidin-1-yl)-5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)phenyl)acetate (7)

[0424]

[0425] Into a 25 mL single-necked bottle, intermediate 6 (230 mg, 0.55 mmol), tetrahydrofuran (3 mL), water (0.6 mL), 5-chloro-2-(piperidin-1-yl)pyrimidine (121 mg, 0.61 mmol) and DIPEA (215 mg, 1.66 mmol) were added in sequence, and the mixture was reacted at 65° C. for 16 h under a nitrogen atmosphere. TLC (V二氯甲烷 :V 甲醇 =20:1) to monitor the complete reaction of the raw materials. The reaction solution was cooled to room temperature, directly mixed, concentrated and purified by rapid preparative chromatography (12 g, V 二氯甲烷 :V 甲醇 =100:1), to obtain 280 mg of white solid intermediate 7, with a yield of 91.5%.

[0426] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 8.87 (s, 2H, ArH), 8.66 (s, 1H, ArH), 7.92 (s, 1H, NH), 7. 37(s,2H,ArH),4.56(br,2H,SO2CH2),3.78(s,2H,CH2COOCH3),3.62(s,3H,CH2COOCH3 ),3.59-3.52(m,2H,NCH2),3.26-3.10(m,3H,NCH2 and CH),2.82(t,J=6.4Hz,2H,SO2CH2C H2CH2),2.31-2.25(m,2H,SO2CH2CH2),2.04-1.99(m,2H,CHCH2),1.70(br,2H,CHCH2).

[0427] Step 7: Synthesis of 2-(2-chloro-4-((2-(4-(5-chloropyrimidin-2-yl)piperidin-1-yl)-5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)phenyl)acetic acid (Example 21)

[0428]

[0429] To a 50 mL single-necked bottle, intermediate 7 (270 mg, 0.48 mmol), methanol (3 mL), THF (3 mL), water (2.2 mL) and sodium hydroxide (89 mg, 2.23 mmol) were added in sequence and reacted at room temperature for 2 h under nitrogen protection. TLC (V 二氯甲烷 :V 甲醇 =20:1) monitor the reaction of the raw materials. Adjust the pH to 4 with dilute hydrochloric acid (1M) under ice bath conditions, solid precipitates, filter, rinse the filter cake with water (20 mL), collect the filter cake and air dry for 16 h to obtain 220 mg of white solid Example 21, with a yield of 93.7%.

[0430] 1H NMR(400MHz,DMSO-d6)δ(ppm):12.53(s,1H,COOH),8.87(s,2H,ArH),8.65(s,1H,ArH ),7.91(s,1H,NH),7.34(s,2H,ArH),4.70-4.55(m,2H,SO2CH2),3.66(s,2H,COCH2), 3.61-3.55(m,2H,NCH2),3.25-3.09(m,3H,NCH2 and CH),2.82(t,J=6.4Hz,2H,SO2CH2CH 2CH2),2.31-2.25(m,2H,SO2CH2CH2),2.07-1.97(m,2H,CHCH2),1.70(s,2H,CHCH2).

[0431] 13 C NMR(101MHz,DMSO-d6)δ(ppm):172.18,170.52,167.34,159.49,156.18,156.14,138.49,133.9 0,132.48,129.01,128.86,121.97,120.23,106.05,51.36,44.09,43.92,38.64,31.92,18.76.

[0432] HRMS(ESI):m / z[M+H] + Theoretical value C 24 H 24 Cl2N6O4S:563.1035; Measured value:563.1025.

[0433] Embodiment 22

[0434] Step 1 Synthesis of 3-ethyl 1-benzyl 2-(2,6-difluoro-4-nitrophenyl)malonate (3)

[0435]

[0436] Sodium hydride (60%, 497 mg, 12.43 mmol) and DMF (10 mL) were added to a 50 mL reaction bottle in sequence. Under nitrogen protection and ice bath conditions, a DMF (2 mL) solution of intermediate 2 (2.51 g, 11.31 mmol) was added dropwise to the system. After the addition was completed, the mixture was kept warm for 0.5 h. A DMF (3 mL) solution of compound 1 (1.00 g, 5.65 mmol) was then added dropwise. The mixture was reacted at 70 °C for 1 h. TLC (V 石油醚 :V 乙酸乙酯=5:1) to monitor the reaction completion. The reaction solution was cooled to an ice bath, quenched with water (30 mL), extracted with ethyl acetate (50 mL × 3), the organic phases were combined, washed with saturated brine (50 mL × 4), the organic phases were collected, dried over anhydrous sodium sulfate, filtered, concentrated and purified by flash chromatography (40 g, V 石油醚 :V 乙酸乙酯 =5:1), to obtain 2.00 g of yellow oily intermediate 3, with a yield of 95.2%.

[0437] 1 H NMR (400MHz, CDCl3) δ (ppm): 7.84 (s, 1H, ArH), 7.82 (s, 1H, ArH), 7.38-7.35 (m, 5H, ArH), 5.19 ( s,2H,ArCH2),5.07(s,1H,CH),4.28-4.21(q,J=7.2Hz,1H,CH3CH2),1.25(t,J=6.4Hz,3H,CH3).

[0438] Step 2 Synthesis of ethyl 2-(4-amino-2,6-difluorophenyl)acetate (4)

[0439]

[0440] In a 50 mL single-necked bottle, intermediate 3 (1.80 g, 4.75 mmol), ethanol (18 mL), ammonium formate (1.51 mg, 23.7 mmol) and Pd / C (10% wet, 270 mg, 0.15 w / w) were added in sequence and reacted at 60°C for 3 h under nitrogen protection. TLC (V 石油醚 :V 乙酸乙酯 =3:1) to monitor the complete reaction of the raw materials. The reaction solution was filtered through diatomaceous earth, the filter cake was rinsed with ethyl acetate (50 mL), the filtrate was collected, concentrated and purified by rapid preparative chromatography (40 g, V 石油醚 :V 乙酸乙酯 =5:1), to obtain 530 mg of white solid intermediate 4, with a yield of 52.0%.

[0441] Step 3 Synthesis of ethyl 2-(4-((2-chloro-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)-2,6-difluorophenyl)acetate (5)

[0442]

[0443] To a 50 mL single-necked bottle, intermediate 4 (530 mg, 2.63 mmol), 1,4-dioxane (10 mL), intermediate 8 of Example 5 (699 mg, 3.16 mmol), sodium carbonate (558 mg, 5.26 mmol), Xantphos (228 mg, 0.40 mmol) and Pd2(dba)3 (180 mg, 0.20 mmol) were added in sequence, and the mixture was reacted at 70°C for 16 h under nitrogen protection. TLC (V 乙酸乙酯 :V 石油醚 =3:1) to monitor the complete reaction of the raw materials. The reaction solution was filtered through diatomaceous earth, the filter cake was rinsed with ethyl acetate (100 mL), the filtrate was collected, concentrated and purified by rapid preparative chromatography (40 g, V 石油醚 :V 乙酸乙酯 =3:1), to obtain 690 mg of white solid intermediate 5, with a yield of 54.2%.

[0444] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 8.84 (s, 1H, NH), 7.43 (d, J = 9.7Hz, 2H, ArH), 4.11 (q, J = 7.2Hz, 2H, CH2CH3), 3.68 (s, 2H, ArCH 2CO),3.19-3.12(m,2H,SCH2),2.81(t,J=6.3Hz,2H,SCH2CH2CH2),2.16-2.10(m,2H,SCH2CH2),1.19(t,J=7.0Hz,3H,CH3).

[0445] Step 4 Synthesis of ethyl 2-(4-((2-chloro-5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)-2,6-difluorophenyl)acetate (6)

[0446]

[0447] To a 100 mL single-necked bottle, intermediate 5 (690 mg, 1.73 mmol), DCM (20 mL) and m-chloroperbenzoic acid (85%, 869 mg, 4.32 mmol) were added in sequence and reacted at room temperature for 2 h under nitrogen protection. TLC (V 乙酸乙酯 :V 石油醚 =2:1) ​​to monitor the complete reaction of the raw material. Add saturated sodium thiosulfate (20 mL) to quench, extract with dichloromethane (50 mL×3), combine the organic phases, wash with saturated brine (50 mL×3), collect the organic phases, dry, filter, concentrate and purify by flash preparative chromatography (20 g, V 二氯甲烷 :V 甲醇=100:1), to obtain 570 mg of white solid intermediate 6, with a yield of 76.5%.

[0448] Step 5 Synthesis of ethyl 2-(4-((2-(4-(5-chloropyrimidin-2-yl)piperidin-1-yl)-5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)-2,6-difluorophenyl)acetate (7)

[0449]

[0450] In a 25 mL single-necked bottle, intermediate 6 (570 mg, 1.32 mmol), tetrahydrofuran (6 mL), water (1 mL), 5-chloro-2-(piperidin-4-yl)pyrimidine (287 mg, 1.45 mmol) and DIPEA (512 mg, 3.96 mmol) were added in sequence and reacted at 65° C. for 16 h under a nitrogen atmosphere. TLC (V 二氯甲烷 :V 甲醇 =20:1) to monitor the complete reaction of the raw materials. The reaction solution was cooled to room temperature, directly mixed, concentrated and purified by rapid preparative chromatography (20 g, V 二氯甲烷 :V 甲醇 =100:1), to obtain 720 mg of white solid intermediate 7, with a yield of 92.1%.

[0451] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 8.87 (s, 2H, ArH), 8.75 (s, 1H, NH), 7.42 (d, J = 9.6Hz, 2H, A rH),4.61(br,2H,SO2CH2),4.10(m,2H,CH2CH3),3.67(s,2H,COCH2),3.59-3.54(m,2H,NC H2),3.23-3.13(m,3H,NCH2 and CH),2.83(t,J=6.4Hz,2H,SCH2CH2CH2),2.31-2.24(m,2H,S CH2CH2),2.07-1.97(m,2H,CHCH2),1.73-1.63(m,2H,CHCH2),1.18(t,J=7.2Hz,3H,CH3).

[0452] Step 6 Synthesis of 2-(4-((2-(4-(5-chloropyrimidin-2-yl)piperidin-1-yl)-5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)-2,6-difluorophenyl)acetic acid (Example 22)

[0453]

[0454] To a 50 mL single-necked bottle, intermediate 7 (300 mg, 0.51 mmol), ethanol (4 mL), THF (4 mL), water (2.2 mL) and sodium hydroxide (89 mg, 2.23 mmol) were added in sequence and reacted at room temperature for 2 h under nitrogen protection. TLC (V 二氯甲烷 :V 甲醇 =20:1) monitor the reaction of the raw materials. Adjust the pH to 4 with dilute hydrochloric acid (1N) under ice bath conditions, solid precipitates, filter, rinse the filter cake with water (20 mL), collect the filter cake and air dry for 16 h to obtain 250 mg of white solid Example 22, with a yield of 87.7%.

[0455] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 12.64 (s, 1H, COOH), 8.86 (d, J = 2.4Hz, 2H, ArH), 8.74 (s, 1H, NH), 7.40 (d, J = 9.4Hz, 2H, ArH), 4.61 (br, 2H, SO2CH2), 3.57 ( s, 4H, NCH2 and COCH2), 3.25-3.14 (m, 3H, NCH2 and CH), 2.83 (br, 2H, SCH2CH2CH2), 2.31-2.26 (m, 2H, SCH2CH2), 2.05-2.01 (m, 2H, CHCH2), 1.70 (s, 2H, CHCH2).

[0456] 13 C NMR (101MHz, DMSO-d6) δ (ppm): 171.34, 170.50, 167.54, 161.03 (dd, J1 = 244.4Hz, J2 = 11.0Hz), 159.41, 156. 13,139.26(t,J=14.2Hz),129.01,106.34,104.56(d,J=29.8Hz),51.36,44.06,43.92,31.93,27.95,18.71.

[0457] HRMS(ESI):m / z[M+H] + Theoretical value C 24 H 23 ClF2N6O4S:565.1236; Measured value:565.1227.

[0458] Embodiment 23

[0459] Step 1 Synthesis of 1-(2-fluorophenyl)cyclopropane-1-carbonitrile (2)

[0460]

[0461] Compound 1 (10.00 g, 74 mmol), 1,2-dibromoethane (27.82 g, 148 mmol), tetrabutylammonium bromide (2.38 g, 7.4 mmol), 50% NaOH (60 mL) and toluene (60 mL) were added to a 250 mL three-necked flask, stirred evenly, and reacted at room temperature overnight under nitrogen protection. TLC (V 石油醚 :V 乙酸乙酯 =5:1) to monitor the complete reaction of the raw materials. Add water (50 mL) to the reaction solution, extract with ethyl acetate (100 mL×2), combine the organic phases, wash with saturated brine (30 mL×2), dry with anhydrous sodium sulfate, filter, concentrate and purify by flash preparative chromatography (120 g, V 石油醚 :V 乙酸乙酯 =10:1), to obtain 5.26 g of colorless oily intermediate 2, with a yield of 44.1%.

[0462] 1 H NMR (400 MHz, CDCl3) δ (ppm): 7.37-7.29 (m, 2H, ArH), 7.16-7.02 (m, 2H, ArH), 1.70-1.67 (m, 2H, cyclopropyl-H), 1.41-1.38 (m, 2H, cyclopropyl-H).

[0463] Step 2 Synthesis of 1-(2-fluoro-4-nitrophenyl)cyclopropane-1-carbonitrile (3)

[0464]

[0465] Intermediate 2 (4.50 g, 28.04 mmol) and sulfuric acid (25 mL) were added to a 250 mL single-necked bottle, and potassium nitrate (3.11 g, 30.84 mmol) was added to an ice-water bath under nitrogen protection, and the reaction was allowed to proceed at room temperature overnight. TLC (V 石油醚 :V 乙酸乙酯 =2:1) ​​to monitor the complete reaction of the raw materials. Add water (50 mL) to the reaction solution, extract with ethyl acetate (100 mL×2), combine the organic phases, dry over anhydrous sodium sulfate, filter, concentrate and purify by flash preparative chromatography (40 g, V 二氯甲烷 :V 甲醇 =50:1), to obtain 6.60 g of white solid intermediate 3, with a yield of 79.4%.

[0466] 1H NMR (400 MHz, DMSO-d6) δ (ppm): 8.30-8.22 (m, 1H, ArH), 8.19 (dd, J1=6.4 Hz, J2=2.9 Hz, 1H, ArH), 7.48 (t, J=9.1 Hz, 1H, ArH), 1.48-1.41 (m, 2H, cyclopropyl-H), 1.10-1.07 (m, 2H, cyclopropyl-H).

[0467] Step 3 Synthesis of 1-(2-fluoro-4-nitrophenyl)cyclopropane-1-carboxylic acid ethyl ester (4)

[0468]

[0469] Intermediate 3 (2.60 g, 12.61 mmol) and ethanol (26 mL) were added to a 250 mL single-necked bottle, and sulfuric acid (5.20 mL) was added under nitrogen protection, and the reaction was refluxed overnight. TLC (V 石油醚 :V 乙酸乙酯 =5:1) to monitor the complete reaction of the raw materials. Add water (50 mL) to the reaction solution, extract with ethyl acetate (150 mL×2), combine the organic phases, dry over anhydrous sodium sulfate, filter, concentrate and purify by flash preparative chromatography (40 g, V 石油醚 :V 乙酸乙酯 =10:1), to obtain 2.50 g of colorless oily intermediate 4, with a yield of 78.3%.

[0470] 1 H NMR (400 MHz, CDCl3) δ (ppm): 8.26-8.12 (m, 2H, ArH) 7.19 (t, J = 8.9 Hz, 1H, ArH), 4.12 (q, J = 7.1 Hz, 2H, CH2), 1.76-1.73 (m, 2H, cyclopropyl-H), 1.25-1.22 (m, 2H, cyclopropyl-H), 1.17 (t, J = 7.1 Hz, 3H, CH3).

[0471] Step 4 Synthesis of 1-(4-amino-2-fluorophenyl)cyclopropane-1-carboxylic acid ethyl ester (5)

[0472]

[0473] In a 100 mL single-necked bottle, intermediate 4 (2.00 g, 7.89 mmol), anhydrous ethanol (40 mL) and 10% wet palladium carbon (400 mg, 20% wt) were added. The reaction was carried out at 60° C. under a hydrogen pressure of 15 psi overnight. TLC (V 石油醚 :V 乙酸乙酯=3:1) to monitor the complete reaction of the raw materials. Ethyl acetate (50 mL) was added to the reaction solution for dilution, filtered through celite, concentrated, and purified by rapid preparative chromatography (20 g, V 石油醚 :V 乙酸乙酯 =5:1), to obtain 1.90 g of colorless oily intermediate 5, with a yield of 86.3%.

[0474] 1 H NMR (400 MHz, CDCl3) δ (ppm): 6.83 (t, J = 8.9 Hz, 1H, ArH), 6.60-6.51 (m, 2H, ArH), 4.10 (q, J = 7.1 Hz, 2H, CH2), 1.62-1.59 (m, 2H, cyclopropyl-H), 1.20-1.13 (m, 5H, cyclopropyl-H and CH3).

[0475] Step 5 Synthesis of ethyl 1-(4-((2-chloro-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)cyclopropane-1-carboxylate (6)

[0476]

[0477] To a 50 mL single-mouth bottle, intermediate 5 (1.00 g, 4.47 mmol), intermediate 8 of Example 5 (1.18 g, 5.37 mmol), XantPhos (388 mg, 0.67 mmol), sodium carbonate (949 mg, 8.96 mmol) and 1,4-dioxane (12 mL) were added in sequence. Under nitrogen protection, tris(dibenzylideneacetone)dipalladium (307 mg, 0.34 mmol) was added and the reaction was carried out at 75°C overnight. TLC (V 二氯甲烷 :V 甲醇 =20:1) to monitor the complete reaction of the raw materials. Add water (100 mL) to the reaction solution, extract with ethyl acetate (200 mL×2), combine the organic phases, wash with saturated brine (30 mL), dry with anhydrous sodium sulfate, filter, concentrate and purify by flash preparative chromatography (20 g, V 石油醚 :V 乙酸乙酯 =5:1), to obtain 850 mg of white solid intermediate 6, with a yield of 46.6%.

[0478] 1H NMR (400 MHz, CDCl3) δ (ppm): 7.53-7.46 (m, 1H, ArH), 7.41 (dd, J1 = 6.4 Hz, J2 = 2.8 Hz, 1H, ArH), 7.03 (t, J = 9.1 Hz, 1H, ArH), 6.56 (s, 1H, NH), 4.11 (q, J = 7.1 Hz, 2H, CH2CH3), 3.21-3.05 (m, 2H, SCH2), 2.88 (t, J = 6.4 Hz, 2H, NCCH2), 2.31-2.21 (m, 2H, SCH2CH2), 1.67-1.65 (m, 2H, cyclopropyl-H), 1.23-1.19 (m, 2H, cyclopropyl-H), 1.17 (t, J = 7.1 Hz, 3H, CH3).

[0479] Step 6 Synthesis of ethyl 1-(4-((2-chloro-5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)cyclopropane-1-carboxylate (7)

[0480]

[0481] Intermediate 6 (850 mg, 2.08 mmol) and dichloromethane (12 mL) were added to a 50 mL single-necked bottle. Under nitrogen protection, m-chloroperbenzoic acid (1.06 g, 5.2 mmol) was added in two batches and reacted at room temperature overnight. TLC (V 石油醚 :V 乙酸乙酯 =1:1) to monitor the complete reaction of the raw materials. Saturated sodium thiosulfate solution (20 mL) was added to the reaction solution and stirred for 10 min, extracted with dichloromethane (100 mL×2), the organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, concentrated and purified by flash preparative chromatography (12 g, V 石油醚 :V 乙酸乙酯 =2:1), to obtain 800 mg of white solid intermediate 7, with a yield of 87.4%.

[0482] Step 7 Synthesis of ethyl 1-(4-((2-(4-(5-chloropyrimidin-2-yl)piperidin-1-yl)-5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)cyclopropane-1-carboxylate (8)

[0483]

[0484] Into a 50 mL single-necked bottle, intermediate 7 (300 mg, 0.68 mmol), tetrahydrofuran (5 mL), water (1 mL), 5-chloro-2-(piperidin-1-yl)pyrimidine (148 mg, 0.75 mmol) and N,N-diisopropylethylamine (264 mg, 2.05 mmol) were added in sequence, and the reaction was carried out at 65 °C under nitrogen protection overnight. TLC (V 二氯甲烷 :V 甲醇 =40:1) to monitor the complete reaction of the raw materials. Water (50 mL) was added to the reaction solution, extracted with ethyl acetate (100 mL×2), the organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, concentrated and purified by flash preparative chromatography (12 g, V 石油醚 :V 乙酸乙酯 =1:2), to obtain 400 mg of white solid intermediate 8, with a yield of 97.7%.

[0485] 1 H NMR (400MHz, CDCl3) δ (ppm): 8.62 (s, 2H, ArH), 8.43 (s, 1H, ArH), 7.51 (dd, J1 = 6.5Hz, J2 = 2.7Hz, 1H, ArH), 7.36-7. 32(m,1H,ArH),6.98(t,J=9.1Hz,1H,NH),4.11(q,J=7.1Hz,2H,CH2CH3),3.40-3.32(m,2H,SCH2),3.19-3.12(m,1 H, CH), 3.02 (t, J = 12.8 Hz, 2H, NCH2), 2.84 (t, J = 6.5 Hz, 2H, NCH2), 2.46-2.43 (m, 2H, NCCH2), 2.06-2.03 (m, 2H, SCH2CH2) 1.90-1.75 (m, 2H, CHCH2), 1.63-1.61 (m, 2H, CHCH2), 1.50-1.43 (m, 2H, cyclopropyl-H) 1.18-1.12 (m, 5H, CH3 and cyclopropyl-H).

[0486] Step 8 Synthesis of 1-(4-((2-(4-(5-chloropyrimidin-2-yl)piperidin-1-yl)-5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)cyclopropane-1-carboxylic acid (Example 23)

[0487]

[0488] To a 50 mL single-necked bottle, intermediate 8 (300 mg, 0.50 mmol), sodium hydroxide (120 mg, 3 mmol), water (3 mL) and ethanol (3 mL) were added in sequence, and the mixture was reacted at 35 °C overnight under nitrogen protection. TLC (V 二氯甲烷 :V 甲醇 =20:1) to monitor the complete reaction of the raw materials. Add water (5 mL) to the reaction solution, adjust the pH to 5 with 1M hydrochloric acid, extract with dichloromethane (50 mL×2), combine the organic phases, wash with saturated brine (30 mL), dry with anhydrous sodium sulfate, filter, concentrate and purify by flash preparative chromatography (20 g, V 石油醚 :V 乙酸乙酯 =1:2), to obtain 38 mg of light yellow solid Example 23, with a yield of 13.3%.

[0489] 1 H NMR (400MHz, CDCl3) δ (ppm): 8.63 (s, 2H, ArH), 8.41 (s, 1H, ArH), 7.54 (dd, J1 = 6.4Hz, J2 = 2.7Hz, 1H, ArH), 7.35-7.31 (m, 1H, ArH), 6.98 (t, J = 9.1Hz, 1H ,NH),4.78(br,2H,SCH2),3.39-3.32(m,2H,NCCH2),3.19-3.12(m,1H,CH), 3.01(t,J=11.7Hz,2H,NCH2),2.83(t,J=6.5Hz,2H,NCH2),2.48-2.40(m,2H SCH2CH2), 2.05-2.02 (m, 2H, CHCH2), 1.82-1.78 (m, 2H, CHCH2), 1.70-1.67 (m, 2H, cyclopropyl-H), 1.26-1.24 (m, 2H, cyclopropyl-H).

[0490] 13 C NMR (101MHz, DMSO-d6) δ (ppm) 174.82, 170.58, 167.02, 159.49, 158.67 (d, J = 244.42Hz), 156.35, 156.12, 134.09 (d, J = 2.0Hz), 128.99, 127.69 ( d,J=15.2Hz), 126(d,J=4.0Hz), 122.65(d,J=8.08Hz), 115.51(d,J=23. 23Hz),105.76,51.46,44.20,43.74,31.92,30.66,24.03,18.80,15.78.

[0491] HRMS(ESI):m / z[M+H]+ Theoretical value C 26 H 26 ClFN6O4S: 573.1487; Measured value: 573.1454 Example 24

[0492] Step 1 Synthesis of 2-(4-((2-(4-(5-chloropyrimidin-2-yl)piperidin-1-yl)-5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)-N-neopentylacetamide (Example 24)

[0493]

[0494] Example 11 (300 mg, 0.55 mmol), anhydrous DMF (12 mL), HATU (316 mg, 0.83 mmol), HOBT (112 mg, 0.83 mmol), DIPEA (213 mg, 1.65 mmol) and t-pentylamine (72 mg, 0.83 mmol) were added to a 100 mL single-necked bottle in sequence, and the mixture was reacted at room temperature for 15 h under nitrogen protection. TLC (V 二氯甲烷 :V 甲醇 =30:1) to monitor the completion of the reaction, add water (50 mL) to quench the reaction system, extract with ethyl acetate (50 mL×3), collect the organic phase, wash with saturated sodium chloride aqueous solution (50 mL×3), dry with anhydrous sodium sulfate, filter, concentrate and perform flash preparative chromatography (12 g, V 石油醚 :V 乙酸乙酯 =1:1) to obtain 208 mg of white solid Example 24 with a yield of 61.4%.

[0495] 1H NMR (400MHz, DMSO-d6) δ (ppm): 8.87 (br, 2H, 2NH), 8.64 (s, 1H, ArH), 7.95 (t, J = 6.3Hz, 1H, ArH), 7.60-7.50 (m, 1H, ArH) ,7.28(t,J=8.4Hz,1H,ArH),7.24-7.16(m,1H,ArH),4.87-4.37(m,2H,NCH2),3.61-3.52(m,2H,NCH2),3.47(s,2H,Hof ArCH2CO),3.25-3.05(m,3H,CH and SO2CH2),2.89(d,J=6.2Hz,2H,NHCH2),2.81(t,J=6.4Hz,2H,SO2CH2CH2CH2),2.34-2.22(m, 2H,SO2CH2CH2CH2),2.07-1.96(m,2H,CHCH2),1.79-1.57(m,2H,CHCH2),0.81(s,9H,3CH3).

[0496] 13 C NMR(101MHz,DMSO-d6)δ(ppm):170.09,169.24,166.80,160.13(d,J C-F =243.4Hz),159.09,155.78,155.68,138.03(d,J C-F =11.1Hz),131.55(d,J C-F =6.1Hz),128.55,118.66(d,J C-F =16.2Hz),116.76(d,J C-F =2.0Hz),108.14(d,J C-F =27.3Hz),105.54,50.90,49.74,43.65,43.42,34.99,32.03,31.44,30.16,27.21,18.31.

[0497] HRMS(ESI):m / z[M+H] + Theoretical value C 29 H 35 ClFN7O3S:616.2273; Measured value:616.2241.

[0498] Embodiment 25

[0499] Step 1 Synthesis of dimethyl 2-(2-fluoro-4-nitrophenyl)malonate (2)

[0500]

[0501] Sodium hydride (60%, 10.00 g, 250.00 mmol) and anhydrous DMF (250 mL) were added to a 1L single-mouth bottle, cooled to 0°C, dimethyl malonate (30.03 g, 227.28 mmol) was added dropwise, and the mixture was reacted at 0°C for 30 min under nitrogen protection. A solution of compound 1 (25.00 g, 113.64 mmol) in anhydrous DMF (120 mL) was then added dropwise, and the mixture was reacted at 70°C for 17 h. TLC (V 石油醚 :V 乙酸乙酯 =4:1) to monitor the completion of the reaction. Add saturated ammonium chloride aqueous solution (1 L) to quench the reaction system, extract with ethyl acetate (1 L × 3), collect the organic phase, wash with saturated sodium chloride aqueous solution (1 L × 3), dry with anhydrous sodium sulfate, filter, concentrate and column chromatography (V 石油醚 :V 乙酸乙酯 =4:1) to obtain 30.60 g of yellow solid intermediate 2 with a yield of 99.3%.

[0502] Step 2 Synthesis of methyl 2-(2-fluoro-4-nitrophenyl)acetate (3)

[0503]

[0504] To a 500 mL single-necked bottle, intermediate 2 (30.59 g, 112.79 mmol), DMSO (210 mL), sodium chloride (6.59 g, 112.79 mmol) and water (4.2 mL) were added in sequence and reacted at 120°C for 4 h under nitrogen protection. TLC (V 石油醚 :V 乙酸乙酯 =10:1) to monitor the completion of the reaction, add water (500 mL) to quench the reaction system, extract with ethyl acetate (500 mL × 3), collect the organic phase, wash with saturated sodium chloride aqueous solution (500 mL × 3), dry with anhydrous sodium sulfate, filter, concentrate and column chromatography (V 石油醚 :V 乙酸乙酯 =10:1) to obtain 8.13 g of yellow oily intermediate 3 in a yield of 33.8%.

[0505] Step 3 Synthesis of 1-(2-fluoro-4-nitrophenyl)cyclobutane-1-carboxylic acid methyl ester (4)

[0506]

[0507] Sodium hydride (60%, 4.95 g, 123.86 mmol) and anhydrous DMF (230 mL) were added to a 500 mL single-mouth bottle, and intermediate 3 (12.00 g, 56.30 mmol) was added under ice bath, and the mixture was reacted at 0°C for 0.5 h. 1,3-diiodopropane (19.99 g, 67.56 mmol) was then added, and the mixture was reacted at 0°C for 4 h under nitrogen protection. TLC (V 石油醚 :V 乙酸乙酯 =10:1) to monitor the completion of the reaction, add saturated ammonium chloride aqueous solution (500 mL) to quench the reaction system, extract with ethyl acetate (500 mL×3), collect the organic phase, wash with saturated sodium chloride aqueous solution (500 mL×3), dry with anhydrous sodium sulfate, filter, concentrate and perform flash preparative chromatography (120 g×2, V 石油醚 :V 乙酸乙酯 =9:1) to obtain 8.53 g of yellow oily intermediate 4 in a yield of 59.8%.

[0508] 1 H NMR (400MHz,CDCl3)δ(ppm):8.07-8.02(m,1H,ArH),7.92-7.85(m,1H,ArH),7.49-7.38(m,1H,ArH),3.69(s,3H,CH3),2.91-2.83(m,2H,CCH2),2.59-2.50(m,2H,CCH2),2.39-2.23(m,1H,H of CCH2CH2),2.02-1.87(m,1H,H of CCH2CH2). Step 4 Synthesis of methyl 1-(4-amino-2-fluorophenyl)cyclobutane-1-carboxylate (5)

[0509]

[0510] In a 250 mL single-necked bottle, intermediate 4 (2.53 g, 9.99 mmol), ethyl acetate (85 mL) and 10% Pd / C (13 wt %, 0.33 g) were added in sequence and reacted at room temperature for 15 h under a hydrogen atmosphere. TLC (V 石油醚 :V 乙酸乙酯 =3:1) to monitor the reaction completion. Filter, concentrate and flash preparative chromatography (40 g, V 石油醚 :V 乙酸乙酯 =4:1) to obtain 1.40 g of yellow oily intermediate 5 with a yield of 62.6%.

[0511] 1H NMR (400MHz, DMSO-d6) δ (ppm): 6.95 (t, J = 8.8Hz, 1H, ArH), 6.34 (d, J = 8.3Hz, 1H, ArH), 6.25 (d, J = 13.4Hz, 1H, A rH),5.29(s,2H,NH2),3.56(s,3H,CH3),2.66-2.55(m,2H,CCH2),2.43-2.29(m,2H,CCH2),2.07-1.92(m,1H,H of CCH2CH2),1.90-1.74(m,1H,H ofCCH2CH2).

[0512] HRMS(ESI):m / z[M+H] + Theoretical value C 12 H 14 FNO2: 224.1087; Found: 224.1051. Step 5 Synthesis of methyl 1-(4-((2-chloro-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)cyclobutane-1-carboxylate (6)

[0513]

[0514] Into a 100 mL single-necked bottle, intermediate 5 (1.89 g, 8.47 mmol), anhydrous 1,4-dioxane (28 mL), intermediate 8 of Example 5 (2.25 g, 10.16 mmol), Pd2(dba)3 (590 mg, 0.64 mmol), XantPhos (730 mg, 1.27 mmol) and sodium carbonate (1.80 g, 16.94 mmol) were added in sequence, and the mixture was reacted at 75°C for 16 h under nitrogen protection. TLC (V 石油醚 :V 乙酸乙酯 =2:1) ​​to monitor the reaction completion, the reaction solution was cooled to room temperature. Filtered, concentrated and subjected to flash preparative chromatography (80 g, V 石油醚 :V 乙酸乙酯 =3:1) to obtain 5.81 g of yellow solid intermediate 6 with a yield of 67.5%.

[0515] 1H NMR(400MHz,DMSO-d6)δ(ppm):8.73(s,1H,NH),7.48-7.27(m,3H,ArH),3.60(s,3H,CH3),3.20-3.08(m,2H,SCH2), 2.84-2.76(m,2H,SCH2CH2CH2),2.75-2.61(m,2H,SCH2CH2CH2),2.49-2.41(m,2H,CCH2),2.20-2.01(m,3H,CCH2,H of CCH2CH2),1.95-1.81(m,1H,H of CCH2CH2).

[0516] HRMS(ESI):m / z[M+H] + Theoretical value C 19 H 19 ClFN3O2S:408.0949; Found:408.0912.

[0517] Step 6 Synthesis of methyl 1-(4-(((2-chloro-5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)cyclobutane-1-carboxylate (7)

[0518]

[0519] To a 250 mL single-necked bottle, intermediate 6 (2.90 g, 7.11 mmol), m-CPBA (85%, 3.61 g, 17.78 mmol) and dichloromethane (87 mL) were added in sequence and reacted at room temperature for 2 h under N2 protection. TLC (V 石油醚 :V 乙酸乙酯 =1:1) monitoring showed that the reaction was complete, saturated sodium thiosulfate aqueous solution (200 mL) was added to quench the reaction system, and dichloromethane (200 mL×3) was used for extraction, and the organic phase was collected. The organic phase was washed with saturated sodium bicarbonate aqueous solution (200 mL×3), dried over anhydrous sodium sulfate, filtered, concentrated and purified by flash chromatography (40 g, V 石油醚 :V 乙酸乙酯 =1:1) to obtain 2.86 g of yellow solid intermediate 7 with a yield of 91.4%.

[0520] 1H NMR(400MHz,DMSO-d6)δ(ppm):9.10(s,1H,NH),7.50-7.36(m,2H,ArH),7.35-7.29(m,1H,ArH),3.75-3.67(m,2H, SCH2),3.61(s,3H,CH3),2.98(t,J=6.3Hz,2H,SO2CH2CH2),2.75-2.64(m,2H,SO2CH2CH2),2.52(d,J=8.2Hz,1H,H of CCH2),2.47(d,J=9.4Hz,1H,H of CCH2),2.39-2.28(m,2H,CCH2),2.16-2.02(m,1H,H of CCH2CH2),1.95-1.81(m,1H,H of CCH2CH2).

[0521] Step 7 Synthesis of methyl 1-(4-((2-(4-(5-chloropyrimidin-2-yl)piperidin-1-yl)-5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)cyclobutane-1-carboxylate (8)

[0522]

[0523] To a 25 mL single-necked bottle, intermediate 7 (500 mg, 0.29 mmol), 5-chloro-2-(piperidin-4-yl)pyrimidine (247 mg, 1.25 mmol), tetrahydrofuran (8.8 mL), water (1.8 mL) and DIPEA (442 mg, 3.42 mmol) were added in sequence, and the mixture was reacted at 65 °C for 16 h under nitrogen protection. TLC (V 二氯甲烷 :V 甲醇 =50:1) to monitor the completion of the reaction, concentrate and perform flash chromatography (12 g, V 二氯甲烷 :V 甲醇 =50:1) to obtain 644 mg of white solid intermediate 8 with a yield of 94.0%.

[0524] 1H NMR (400MHz, DMSO-d6) δ (ppm): 8.87 (s, 2H, ArH), 8.68 (s, 1H, NH), 7.57 (d, J = 13.2Hz, 1 H,ArH),7.42-7.07(m,2H,ArH),4.95-4.28(m,2H,SO2CH2),3.67-3.48(m,5H,NCH2and CH3),3.24-3.10(m,3H,NCH2 and CH),2.92-2.77(m,2H,SO2CH2CH2CH2),2.49-2.38(m,2H,SO2CH2CH2CH2),2.36-2.22(m,2H,CCH2),2.09-2.02(m,3H,CCH2,H of CHCH2),1.93-1.81(m,1H,H of CHCH2),1.80-1.58(m,2H,CHCH2),1.33-1.22(m,2H,CCH2CH2).

[0525] Step 8 Synthesis of 1-(4-((2-(4-(5-chloropyrimidin-2-yl)piperidin-1-yl)-5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)cyclobutane-1-carboxylic acid (Example 25)

[0526]

[0527] To a 50 mL single-necked bottle, intermediate 8 (344 mg, 0.57 mmol), ethanol (6 mL), tetrahydrofuran (12 mL), water (6 mL) and sodium hydroxide (114 mg, 2.85 mmol) were added in sequence and reacted at 35 °C for 1.5 h under nitrogen protection. TLC (V 二氯甲烷 :V 甲醇 =50:1) monitor the remaining raw material to about 10%, adjust the pH to 7 with 1N dilute hydrochloric acid under ice bath, and concentrate to remove the organic solvent. Then adjust the pH to 3 with 1N dilute hydrochloric acid, filter, collect the filter cake, and then pass it through a thick preparation plate (V 二氯甲烷 :V 甲醇 =19:1) to obtain 71 mg of white solid Example 25 with a yield of 21.3%.

[0528] 1H NMR(400MHz,DMSO-d6)δ(ppm):12.37(s,1H,COOH),8.87(s,2H,ArH),8.67(s,1H,NH),7 .60-7.51(m,1H,ArH),7.35-7.22(m,2H,ArH),4.63(br,2H,SO2CH2),3.59-3.52(m,2H,N CH2),3.27-3.09(m,3H,CH and NCH2),2.82(t,J=6.4Hz,2H,SO2CH2CH2CH2),2.71-2.61(m,2 H,SO2CH2CH2CH2),2.47-2.38(m,2H,CCH2),2.33-2.23(m,2H,CCH2),2.15-1.99(m,3H,H ofCHCH2 and CHCH2),1.88-1.80(m,1H,H of CHCH2),1.77-1.66(m,2H,CCH2CH2).

[0529] 13 C NMR(101MHz,DMSO-d6)δ(ppm):175.87,170.10,166.83,159.82(d,J C-F =244.4Hz),159.10,155.72,155.67,138.22(d,J C-F =11.1Hz),128.56,128.20(d,J C-F =6.1Hz),126.07(d,J C-F =15.2Hz),116.22(d,J C-F =3.0Hz),108.15(d,J C-F =27.3Hz),105.60,50.90,48.37,43.65,43.49,31.46,31.06,30.18,18.31,16.83.

[0530] HRMS(ESI):m / z[M+H] + Theoretical value C 27 H 28 ClFN6O4S:587.1643; Found:587.1639.

[0531] Embodiment 26

[0532] Step 1 Synthesis of methyl 1-(4-((2-(4-(5-chloropyrimidin-2-yl)-3,6-dihydropyridin-1(2H)-yl)-5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)cyclobutane-1-carboxylate (2)

[0533]

[0534] In a 25 mL single-necked bottle, intermediate 7 of Example 25 (210 mg, 0.53 mmol), intermediate 3 of Example 1 (114 mg, 0.58 mmol), DIPEA (205 g, 1.59 mmol), THF (4 mL) and H2O (1 mL) were added in sequence, and the mixture was reacted at 65°C overnight under nitrogen protection. TLC (V 二氯甲烷 :V 甲醇 =30:1) The reaction was monitored to be complete, concentrated, and slurried with tetrahydrofuran (1 mL) to obtain 130 mg of white solid intermediate 2 with a yield of 87.6%.

[0535] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 8.91 (s, 2H, ArH), 8.72 (s, 1H, NH), 7.60 (d, J = 13.0Hz, 1H, ArH), 7.39 (d, J =5.2Hz,2H,ArH),7.30(s,1H,C=CH),4.48(br,2H,SO2CH2),3.99(br,2H,NCH2),3.64-3.57(m,3H,NCH2,H of NCH2CH2),3.62(s,3H,CH3),3.16-3.11(m,1H,H of NCH2CH2),2.86(t,J=6.5Hz,2H,SO2CH2CH2CH2),2.74-2.67(m,4H,CH2CH2CH2),2.33-2.27(m,2H,SO2CH2CH2CH2),2.14-2.07(m,1H,H of CH2CH2CH2),1.94-1.86(m,1H,H ofCH2CH2CH2).

[0536] Step 2 Synthesis of 1-(4-((2-(4-(5-chloropyrimidin-2-yl)-3,6-dihydropyridin-1(2H)-yl)-5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)cyclobutane-1-carboxylic acid (Example 26)

[0537]

[0538] To a 25 mL single-necked bottle, intermediate 2 (280 mg, 0.47 mmol), acetic acid (5 mL) and concentrated hydrochloric acid (2.5 mL) were added in sequence and reacted at 90°C for 40 min under nitrogen protection. TLC (V 二氯甲烷 :V 甲醇 =15:1) monitor the reaction completion, concentrate to (4mL), add ice water (8mL) to the residue, stir at room temperature for 1h. Filter, rinse the filter cake with water (2mL×3), and collect the filter cake. Prepare plate purification (V 二氯甲烷 :V 甲醇 =20:1) three times to obtain 49 mg of white solid Example 26, with a yield of 17.8%.

[0539] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 12.39 (s, 1H, COOH), 8.90 (s, 2H, ArH), 8.70 (s, 1H, NH), 7.58 (d, J = 1 3.0Hz,1H,ArH),7.34-7.33(m,2H,ArH),7.31-7.28(m,1H,C=CH),4.47(br,2H,SO2CH2CH2CH2),3.9 8(br,2H,NCH2),3.59-3.56(m,2H,NCH2CH2),2.85(t,J=6.4Hz,2H,SO2CH2CH2CH2),2.72-2.65(m, 4H,CH2CH2CH2),2.47-2.42(m,2H,NCH2CH2),2.32-2.26(m,2H,SO2CH2CH2CH2),2.14-2.07(m,1H,H of CH2CH2CH2),1.89-1.85(m,1H,H ofCH2CH2CH2).

[0540] 13 C NMR(101MHz,DMSO-d6)δ(ppm):176.35,167.12,161.90,160.11(d,J C-F =245.4Hz),159.55,156.06,155.88,138.23(d,J C-F =11.1Hz),134.03,128.85,128.41(d,J C-F =6.0Hz),126.58(d,J C-F =15.1Hz),116.74,108.64(d,J C-F=27.2Hz),106.44,67.47,55.36,51.33,48.84,44.66,31.89,31.52,25.21,18.74,17.30.

[0541] HRMS(ESI):m / z[M+H] + Theoretical value C 27 H 26 ClFN6O4S: 585.1487; Measured value: 585.1507 Example 27

[0542] Step 1 Synthesis of methyl 2-(4-((2-(4-(5-chloropyrimidin-2-yl)piperazin-1-yl)-5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)acetate (1)

[0543]

[0544] In a 25 mL single-necked bottle, intermediate 3 of Example 11 (200 mg, 0.50 mmol), 5-chloro-2-(piperazin-1-yl)pyrimidine (109 mg, 0.55 mmol), DIPEA (194 mg, 1.50 mmol), THF (5 mL) and H2O (1 mL) were added in sequence, and the mixture was reacted at 65°C overnight under nitrogen protection. TLC (V 二氯甲烷 :V 甲醇 =50:1) The reaction was monitored to be complete, concentrated, and slurried with tetrahydrofuran (2 mL) to obtain 142 mg of white solid intermediate 1 with a yield of 87.6%.

[0545] Step 2 Synthesis of 2-(4-((2-(4-(5-chloropyrimidin-2-yl)piperazin-1-yl)-5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)amino)-2-fluorophenyl)acetic acid (Example 27)

[0546]

[0547] To a 25 mL single-necked bottle, intermediate 1 (142 mg, 0.25 mmol), ethanol (2 mL), tetrahydrofuran (2 mL), water (2 mL) and sodium hydroxide (15 mg, 0.38 mmol) were added in sequence and reacted at 30 °C for 4 h under N2 protection. TLC (V 二氯甲烷 :V 甲醇 =50:1) monitoring the completion of the reaction, adjusting the pH of the solution to 7 with 3M hydrochloric acid, concentrating, continuing to adjust the pH of the solution to 4 with 3M hydrochloric acid, filtering, washing the filter cake with water (10 mL), and drying the filter cake to obtain 120 mg of white solid Example 27, with a yield of 87.6%.

[0548] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 12.62 (s, 1H, COOH), 8.70 (s, 1H, NH), 8.46 (s, 2H, ArH), 7.56 (dd, J1 = 12.1Hz, J2 = 2.1Hz, 1H, ArH), 7.36-7.24 (m ,2H,ArH),3.82-3.80(m,8H,piperazine-H),3.63-3.56(m,4H,CH2COOH,SO2CH2),2.84(t,J=6.4Hz,2H,SO2CH2CH2CH2),2.35-2.25(m,2H,SO2CH2CH2).

[0549] 13 C NMR(101MHz,DMSO)δ(ppm):172.35,167.22,160.74(d,J C-F =244.4Hz),159.87,159.63,156.49,156.19,138.70(d,J C-F =11.1Hz),132.31(d,J C-F =6.1Hz),118.20,118.08(d,J C-F =17.2Hz),117.37(d,J C-F =3.0Hz),108.66(d,J C-F =27.3Hz),106.42,51.35,43.72,43.50,34.44,31.89,18.75.

[0550] HRMS(ESI):m / z[M+H] + Theoretical value C 23 H 23 ClFN7O4S:548.1283; Found:548.1215.

[0551] Biological Activity Study (I) Inhibitory Activity of the Compounds of the Invention on PDE4B Enzyme

[0552] 1. Materials and Instruments

[0553]

[0554] 2 Experimental process

[0555] 2.1 Compound preparation and treatment: All compounds were prepared into 50 mM stock solutions in dimethyl sulfoxide.

[0556] 2.2 Preparation of working stock solutions: a) Reference Rolipram was serially diluted 3-fold starting from 2 mmol with dimethyl sulfoxide for a total of 10 concentrations. b) 2 mmol of the compound of the invention was serially diluted 3-fold with dimethyl sulfoxide (DMSO) for a total of 10 concentrations. c) Prepare 200X positive control (2 mM Rolipram) and 200X vehicle control (100% dimethyl sulfoxide). d) The compound plate was centrifuged at 1000 rpm for 1 minute.

[0557] 2.3 Complex screening

[0558] a) Using the Echo 550, transfer 20 nL of compound dilution to each well of the assay plate.

[0559] b) Seal the test plate and centrifuge at 1000 rpm for 1 minute.

[0560] c) Prepare 2X PDE4B2 in chilled PDE Assay Buffer.

[0561] d) Add 2 μL of 2X PDE4B2 to each well of the assay plate (prepared in step b).

[0562] e) Seal the assay plate and equilibrate at RT for 10 minutes.

[0563] f) Prepare 2X Cyclic-3',5'-AMP in PDE Assay Buffer.

[0564] g) Start the reaction by adding 2 μl of 2X Cyclic-3',5'-AMP (prepared in step f) to each well of the assay plate (prepared in step e). Incubate at room temperature for 60 minutes.

[0565] h) Add 4 μL AMP-Glo ​​Reagent I.

[0566] i) Add 8 μL AMP detection solution and incubate at room temperature for 60 minutes.

[0567] k) Read the RLU signal on an Envision 2105 plate reader.

[0568] 3. Test results

[0569] As shown in the table below, the compounds of the present invention have extremely strong PDE4B inhibitory activity, and the activity of some compounds is much stronger than the inhibitory activity of compound II disclosed in WO2013026797.

[0570]

[0571]

[0572] Note: “+”: IC 50 >1μM

[0573] "++" IC 50 =100nM~1μM

[0574] "+++" IC 50 =20nM~100nM

[0575] "++++"IC 50 =1~20nM

[0576] "++++++" IC 50 <1nM

[0577] (II) Selectivity of the compounds of the present invention for PDE4 enzyme subtypes 1. Materials and instruments

[0578]

[0579] 2. Methods

[0580] Same as PDE4B detection method

[0581] 3. Results

[0582] The compound of the present invention has better selectivity for PDE4B and is superior to compound 2 disclosed in patent WO2013026797.

[0583]

[0584] (III) Toxicity of the compounds of the present invention to hERG

[0585] HEK-293 cells stably expressing hERG potassium channel were used (purchased from Creacell: Product No. A-0320). HEK293 cells were cultured in DMEM medium containing 10% fetal bovine serum and 0.8 mg / mL G418 at 37°C and 5% carbon dioxide concentration. Before the patch clamp test, cells were stained with TrypLE TM Express separation, 4×10 3 The cells were plated onto cover slips and cultured in 24-well plates (final volume: 500 μL) and assayed 18 hours later.

[0586] Whole-cell patch clamp detection: After the whole-cell seal is formed, the cell membrane voltage is clamped at -80mV. The clamp voltage is also depolarized from -80mV to -50mV for 0.5s (as a leakage current detection), then stepped to 30mV for 2.5s, and then quickly restored to -50mV for 4s to stimulate the tail current of the hERG channel. Repeat data collection every 10s to observe the effects of drugs on the hERG tail current. A 0.5s -50mV stimulation is used as a leakage current detection. The experimental data is collected by the EPC-10 amplifier and stored in the software. When the hERG current recorded by the whole cell is stable, the drug is administered. After each drug concentration acts for 5min (or the current is stable), the next concentration is detected. Each test compound is tested (0.03μM, 0.3μM, 1μM, 3μM, 10μM, 30μM) The data at the concentration, and the hERG IC is calculated according to the calculation formula. 50 value.

[0587] Compared with compound II disclosed in WO2013026797, the activity of the compounds of the present invention in inhibiting hERG potassium channel is significantly reduced, and most compounds inhibit hERG with IC 50 >30μM, with low risk of cardiotoxicity and higher safety.

[0588]

[0589]

[0590] (IV) Effects of the compounds of the present invention on chronic obstructive pulmonary disease in rats

[0591] 1. Animals and Reagents

[0592]

[0593] 2. Experimental Procedure

[0594] 2.1 Experimental Grouping

[0595] 40 SD rats were adaptively raised in an animal experimental center for 7 days, with 12-hour alternating lighting, an ambient temperature of 20-25°C, and a humidity of 50%-65%. Subsequently, the SD rats were randomly divided into 5 groups according to body weight, with 8 rats in each group, half of which were male and half were female, and they were respectively a blank control group, a model control group, and an Example 11 administration group (doses were 5, 10, and 20 mg / kg, respectively).

[0596] 2.2 Preparation of experimental reagents

[0597] Sample solution to be tested: 5 mg, 10 mg and 20 mg of the compound of Example 11 were weighed and dissolved in 10 ml of 0.5% CMC-Na aqueous solution to prepare solutions with concentrations of 0.5, 1 and 2 mg / ml.

[0598] 2.3 Experimental plan

[0599] The SD rats except the blank control group were fumigated + intratracheally instilled with LPS to establish a rat model of chronic obstructive pulmonary disease (COPD). On the 1st, 14th, 28th, and 42nd day of modeling, the rats were anesthetized with chloral hydrate, and then 125μLLS (2mg / mL) solution was instilled into the trachea. The animals were placed in a homemade fumigation box for the rest of the time. Each group of rats was fumigated with 10 lit cigarettes for 1.5 hours (once in the morning and once in the afternoon) for 50 consecutive days. After the modeling was completed, each group of rats was given medication for 28 consecutive days, twice a day.

[0600] 2.4 Experimental Results

[0601] 2.4.1 Results of forced vital capacity (FVC) and forced expiratory volume in 1 second (FEV1) in COPD rats

[0602] The rat pulmonary function tester was used to detect the pulmonary function FVC and FEV1 on the 14th day of treatment and the last dose, and the FEV1 / FVC ratio was calculated. The results are shown in Table 1.

[0603] Table 1. Effect of Example 11 on FEV1 / FVC in rats with chronic pulmonary obstructive pulmonary disease (Mean±SD, n=8)

[0604]

[0605] ## P<0.01vs blank group; * P<0.05, ** P<0.01vs model group.

[0606] Compared with the blank group, the FEV1 / FVC (percentage of maximum vital capacity in 1 second) of the rats in the model group was significantly reduced (P<0.01), indicating that the model was successfully established; compared with the model group, the group administered with the compound of Example 11 could significantly improve the FEV1 / FVC lung function of the rats (P<0.05, P<0.01), indicating that it had a good therapeutic effect; the 10 mg / kg and 20 mg / kg dosage administration groups of the compound of Example 11 were significantly better than the 5 mg / kg dosage administration group (P<0.01), indicating that there was a dose gradient relationship.

[0607] 2.4.2 Levels of IL-1β and TNF-α in serum of COPD rats

[0608] After the last administration, blood was collected for determination of inflammatory indexes, and the levels of IL-1β and TNF-α in serum were detected according to the ELISA instructions. The results are shown in Table 2.

[0609] Table 2. Effect of Example 11 on IL-1β and TNF-α in serum of COPD rats (mean ± SD, n = 8)

[0610]

[0611] ## P<0.01vs blank group; * P<0.05, ** P<0.01vs model group.

[0612] Compared with the blank group, the levels of IL-1β and TNF-α in the serum of the rats in the model group were significantly increased (P<0.01), indicating that the model was successfully established; compared with the model group, the group administered with the compound of Example 11 could significantly improve the levels of IL-1β and TNF-α in the serum of the rats (P<0.01), indicating that it had a significant therapeutic effect; and a dose gradient relationship was suggested.

[0613] 2.4.3 Results of lung tissue pathology in COPD rats

[0614] After the last treatment, all three lobes of the right lung of the rats were removed, fixed in 10% formaldehyde, dehydrated, embedded in paraffin, and sliced. HE staining was performed to observe the morphological changes of the lung tissues of the rats in each group. The results are as follows: Figure 1 And as shown in Table 3.

[0615] Table 3. Effect of Example 11 on lung tissue pathology scores in COPD rats (mean ± SD, n = 8)

[0616]

[0617] ## P<0.01vs blank group; * P<0.05, ** P<0.01vs model group.

[0618] Compared with the blank group, the alveolar septa in the lung tissue of the rats in the model group were widened and inflammatory cells infiltrated, and the adjacent lung tissue and alveolar cavity showed compensatory expansion, indicating that the model was successful; compared with the model group, the group treated with the compound of Example 11 could significantly improve the pathological score of the rat lung tissue (P<0.01), indicating a significant therapeutic effect; and suggesting a dose gradient relationship.

[0619] (V) Effects of the compounds of the present invention on eosinophilic asthma in mice induced by house dust mite extract 1. Animals and reagents

[0620]

[0621] 2. Experimental Procedure

[0622] 2.1 Experimental Grouping

[0623] Fifty BALB / c mice were randomly divided into five groups according to body weight, with 10 mice in each group, half of which were male and half were female, including a blank control group, a model control group, and an Example 11 medication group (doses were 5, 10, and 20 mg / kg, respectively).

[0624] 2.2 Preparation of experimental reagents

[0625] House dust mite extract solution: Weigh 25 mg of house dust mite extract, add it into 1 ml of normal saline and mix well to prepare a solution with a concentration of 25 mg / ml.

[0626] Sample solution to be tested: 5 mg, 10 mg and 20 mg of the compound of Example 11 were respectively dissolved in 10 ml of 0.5% CMC-Na aqueous solution to prepare solutions with concentrations of 0.5, 1 and 2 mg / ml.

[0627] 2.3 Experimental plan

[0628] BALB / c mice other than the blank control group were sensitized by intranasal drops of house dust mite extract solution on the 1st, 7th and 14th days of modeling. After the mice were anesthetized with chloral hydrate, 25mg / ml house dust mite extract solution was slowly dripped into the nose (20ul); on the 20th to 25th day, 50mg / ml house dust mite extract solution was dripped into the nose for 6 consecutive days (20ul) to establish the mouse eosinophilic asthma model. After that, each treatment group was treated with continuous medication for 14 days, twice a day.

[0629] 2.4 Experimental Results

[0630] 2.4.1 Results of eosinophil count and sneezing frequency

[0631] The number of eosinophils in the lung lavage fluid of mice and the number of sneezing times of mice within 20 minutes after the last administration were counted. The results are shown in Table 4.

[0632] Table 4. Effects of Example 11 on eosinophils and sneezing frequency in eosinophilic asthma mice (mean ± SD, n = 10)

[0633]

[0634] ## P<0.01vs blank group; * P<0.05, ** P<0.01vs model group.

[0635] Compared with the blank group, the percentage of eosinophils in the alveolar lavage fluid of the model group was significantly increased (P<0.01), indicating that the model was successfully established; compared with the model group, the group treated with the compound of Example 11 could significantly reduce eosinophils (P<0.05, P<0.01), having a significant therapeutic effect; and suggesting a dose gradient relationship.

[0636] Compared with the blank group, the number of sneezes in the model group within 20 minutes was significantly increased (P<0.01), indicating that the model was successfully established; compared with the model group, the groups treated with the compound of Example 11 were able to significantly reduce the number of sneezes within 20 minutes (P<0.01), showing a significant therapeutic effect; and suggesting a dose gradient relationship.

[0637] 2.4.2 Contents of IgE and IL-4 in bronchoalveolar lavage fluid

[0638] The levels of IgE and IL-4 in the bronchoalveolar lavage fluid of mice with eosinophilic asthma were measured. The results are shown in Table 5.

[0639] Table 5. Effect of Example 11 on IgE and IL-4 in bronchoalveolar lavage fluid of mice with eosinophilic asthma (Mean±SD, n=10)

[0640]

[0641] ## P<0.01vs blank group; * P<0.05, ** P<0.01vs model group.

[0642] Compared with the blank group, the IgE content in the alveolar lavage fluid of the model group mice was significantly increased (P<0.01), indicating that the model was successfully established; compared with the model group, the Example 11 compound-administered groups were able to significantly reduce the IgE content in the alveolar lavage fluid (P<0.05, P<0.01), having a significant therapeutic effect; and suggesting a dose gradient relationship.

[0643] Compared with the blank group, the IL-4 content in the alveolar lavage fluid of the model group mice was significantly increased (P<0.01), indicating that the model was successfully established; compared with the model group, the Example 11 compound-administered groups were able to significantly reduce the IL-4 content in the alveolar lavage fluid (P<0.01), having a significant therapeutic effect; and suggesting a dose gradient relationship.

[0644] (VI) Effects of the compounds of the present invention on bleomycin-induced pulmonary fibrosis in rats

[0645] 1. Animals and Reagents

[0646]

[0647] 2. Experimental Procedure

[0648] 2.1 Experimental Grouping

[0649] Fifty SD rats were randomly divided into five groups according to their body weight, with 10 rats in each group, half of which were male and half were female, including a blank control group, a model control group, and an Example 11 medication group (doses were 5, 10, and 20 mg / kg, respectively).

[0650] 2.2 Preparation of experimental reagents

[0651] Bleomycin solution: Weigh 5 mg of bleomycin powder and add it to 1 ml of normal saline and mix well to prepare a solution with a concentration of 5 mg / ml.

[0652] Sample solution to be tested: 5 mg, 10 mg and 20 mg of the compound of Example 11 were weighed and dissolved in 10 ml of 0.5% CMC-Na aqueous solution to prepare solutions with concentrations of 0.5, 1 and 2 mg / ml.

[0653] 2.3 Experimental plan

[0654] After rats were anesthetized with chloral hydrate, bleomycin solution was injected into the trachea through the vocal cords, the dose was 5 mg / kg, and the administration volume was 1 ml / kg; the blank group rats were injected with an equal amount of normal saline into the trachea. The time for intratracheal injection of bleomycin was set at the first day, and oral administration began on the 8th day, with an administration volume of 10 ml / kg, twice a day, for 14 consecutive days. Blood and lungs were collected from animals in each group on the 22nd day. After the rats were killed, the trachea was exposed, and the intravenous cannula was inserted into the rat bronchi. PBS (5 ml) was lavaged twice, and the lavage fluid was collected. Then, centrifuged at 1500 r / min for 10 minutes. The supernatant was collected.

[0655] 2.4 Experimental Results

[0656] The degree of fibrosis was determined by Masson's staining; meanwhile, ELISA was used to measure the levels of IL-6, TNF-α, and TGF-β1 in bronchoalveolar lavage fluid.

[0657] Table 6. Effect of Example 11 on bleomycin-induced pulmonary fibrosis in rats (mean ± SD, n = 10)

[0658]

[0659] ## P<0.01vs blank group; * P<0.05, ** P<0.01vs model group.

[0660] like Figure 2As shown in Table 6, compared with the blank control group, the model group rats can significantly increase the collagen fiber content of lung tissue after bleomycin induction (P<0.01), indicating that the modeling is successful; compared with the model control group, the Example 11 administration group can significantly reduce the collagen fiber content of lung tissue (P<0.05, P<0.01), indicating that the administration group has a therapeutic effect.

[0661] Table 7. Effect of Example 11 on TGF-β and IL-6 in rat bronchoalveolar lavage fluid induced by bleomycin (mean ± SD, n = 10)

[0662]

[0663] # P<0.05, ## P<0.01vs blank group; * P<0.05, ** P<0.01vs model group.

[0664] As shown in Table 7, compared with the blank control group, the model group rats could significantly increase the content of TGF-β and IL-6 in the alveolar lavage fluid after bleomycin induction (P<0.01), indicating that the model was successfully established; compared with the model control group, the Example 11 administration group could significantly reduce the content of TGF-β and IL-6 in the alveolar lavage fluid (P<0.05, P<0.01), indicating that the administration group had a therapeutic effect.

[0665] The references mentioned herein are all incorporated herein by reference. It should be understood that many changes and modifications can be made to the technical solutions of the present invention without departing from the spirit and scope of the present disclosure.

Claims

1. Compound represented by formula (I) or its stereoisomers, tautomers, solvates, prodrugs, isotope labels, and pharmaceutically acceptable salts thereof, wherein X is selected from S=O, S(=O)2; R1 is selected from H, C1-C6 alkyl; R1' is selected from C3-C8 carbocyclic group, C6-C 10 aryl, 5-10 membered heteroaryl, 5-10 membered heterocyclic group; wherein the carbocyclic group, aryl, heteroaryl, heterocyclic group is replaced by R4-(CR a R b ) m -substituted and optionally substituted by halogen, hydroxy, amino, mercapto, C1-C6 alkyl, C1-C6 alkoxy; R2, R2', R3, R3' are each independently selected from hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl; R4 is selected from hydroxyl, amino, thiol, carboxyl, -SO3H, -PO4H, tetrazolyl, triazolyl, -C(=O)OC1-C6 alkyl, -C(=O)NR c R d ; R a , R b are each independently selected from hydrogen, C1-C6 alkyl, halogen, hydroxyl, or R a , R b Together with the carbon atom to which it is attached, it forms a C3-C6 carbocyclic group; R c , R d are each independently selected from hydrogen, C1-C6 alkyl, or R c , R d Together with the nitrogen atom to which it is attached, it forms a 3- to 6-membered heterocyclic group; Het is selected from 5- or 6-membered nitrogen aromatic group; L is selected from a bond, C1-C6 alkylene, -C(=O)-, -NR e C(=O)-、-C(=O)-N(R e )-, -OC(=O)-, -C(=O)O-; R e Selected from hydrogen or C1-C6 alkyl; Ar is selected from 5-10 membered heterocyclic group, 5-10 membered heteroaryl, C6-C 10 Aryl, C3-C8 carbocyclic group; the heterocyclic group, heteroaryl group, aryl group, carbocyclic group are optionally substituted by one or more groups selected from the following: halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy; n is selected from 1 or 2; m is selected from 1, 2, 3, 4, 5 or 6; The condition is: when X is S=O, Het is When R1' is a C3-C8 carbocyclic group, L is not a bond.

2. The compound according to claim 1 or its stereoisomers, tautomers, solvates, prodrugs, isotope-labeled substances, and pharmaceutically acceptable salts thereof, wherein X is selected from S=O, S(=O)2; R1 is selected from H, C1-C6 alkyl; R1' is selected from C3-C8 carbocyclic group, C6-C 10 aryl, 5-10 membered heteroaryl; wherein the carbocyclic group, aryl, heteroaryl is replaced by R4-(CR a R b ) m -substituted and optionally substituted by halogen, hydroxy, amino, mercapto, C1-C6 alkyl, C1-C6 alkoxy; R2, R2', R3, R3' are each independently selected from hydrogen, C1-C6 alkyl; R4 is selected from hydroxyl, amino, thiol, carboxyl, -SO3H, -PO4H, tetrazolyl, triazolyl, -C(=O)OC1-C6 alkyl, -C(=O)NR c R d ; R a , R b are each independently selected from hydrogen, C1-C6 alkyl, halogen, hydroxyl, or R a , R b Together with the carbon atom to which it is attached, it forms a C3-C6 carbocyclic group; R c , R d are each independently selected from hydrogen, C1-C6 alkyl, or R c , R d Together with the nitrogen atom to which it is attached, it forms a 3- to 6-membered heterocyclic group; Het is selected from 5- or 6-membered nitrogen aromatic group; L is selected from a bond, C1-C6 alkylene, -C(=O)-, -NR e C(=O)-、-C(=O)-N(R e )-, -OC(=O)-, -C(=O)O-; R e Selected from hydrogen or C1-C6 alkyl; Ar is selected from 5-10 membered heteroaryl, C6-C 10 Aryl; the heteroaryl and aryl are optionally substituted by one or more groups selected from the following: halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy; n is selected from 1 or 2; m is selected from 1, 2, 3, 4, 5 or 6; The condition is: when X is S=O, Het is When R1' is a C3-C8 carbocyclic group, L is not a bond.

3. The compound according to claim 1 or 2, or its stereoisomers, tautomers, solvates, prodrugs, isotope-labeled substances, and pharmaceutically acceptable salts thereof, wherein R1' is selected from cyclopropane, cyclobutane, cyclopentane, cyclohexane, phenyl, pyrrolyl, furanyl, thienyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl; the above groups are replaced by R4-(CR a R b ) m -substituted and optionally substituted by halogen, hydroxy, amino, mercapto, C1-C6 alkyl, C1-C6 alkoxy; R2, R2', R3, R3' are each independently selected from hydrogen, C1-C6 alkyl; R4 is selected from hydroxyl, amino, thiol, carboxyl, -SO3H, -PO4H, tetrazolyl, triazolyl, -C(=O)OC1-C 20 Alkyl, -C(=O)NR c R d ; R a , R b are each independently selected from hydrogen, C1-C6 alkyl, halogen, hydroxyl, or R a , R b Together with the carbon atom to which it is attached, it forms a C3-C6 carbocyclic group; R c , R d are each independently selected from hydrogen, C1-C6 alkyl, or R c , R d Together with the nitrogen atom to which it is attached, it forms a 3- to 6-membered heterocyclic group; Het is selected from 5- or 6-membered nitrogen aromatic group; L is selected from a bond, C1-C6 alkylene, -C(=O)-, -NR a C(=O)-、-C(=O)-N(R a )-, -OC(=O)-, -C(=O)O-; Ar is selected from pyrrolyl, furanyl, thienyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, phenyl; the above groups are optionally substituted by one or more groups selected from the following: halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy; n is selected from 1 or 2; m is selected from 1, 2, 3, 4, 5 or 6; The condition is: when X is S=O, Het is When R1' is cyclopropane, cyclobutane, cyclopentane or cyclohexane, L is not a bond.

4. The compound according to any one of claims 1 to 3 or its stereoisomers, tautomers, solvates, prodrugs, isotope-labeled substances, and pharmaceutically acceptable salts thereof, wherein R1' is selected from cyclobutane, phenyl, pyridyl; the above groups are replaced by R4-(CR a R b ) m -substituted and optionally substituted by halogen; R2, R2', R3, R3' are each independently selected from hydrogen, C1-C6 alkyl; R4 is selected from hydroxyl, amino, thiol, carboxyl, -SO3H, -PO4H, tetrazolyl, triazolyl, -C(=O)OC1-C 20 Alkyl, -C(=O)NR c R d ; R a , R b are each independently selected from hydrogen, C1-C3 alkyl, halogen, or R a , R b Together with the carbon atom to which it is connected, it forms a cyclopropane group or a cyclobutane group; R c , R d are each independently selected from hydrogen, C1-C3 alkyl, or R c , R d Together with the nitrogen atom to which it is attached, it forms a 3- to 6-membered heterocyclic group; Het is selected from 5- or 6-membered nitrogen aromatic group; L is selected from a bond, C1-C6 alkylene, -C(=O)-, -NR a C(=O)-、-C(=O)-N(R a )-, -OC(=O)-, -C(=O)O-; Ar is selected from pyrrolyl, furanyl, thienyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, phenyl; the above groups are optionally substituted by one or more groups selected from the following: halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy; n is selected from 1 or 2; m is selected from 1, 2, 3, 4, 5 or 6; The condition is: when X is S=O, Het is When R1' is cyclobutane, L is not a bond.

5. The compound according to any one of claims 1 to 4, which is a compound represented by formula (II): or its stereoisomers, tautomers, solvates, prodrugs, isotope-labeled substances, and pharmaceutically acceptable salts thereof, wherein: X is selected from S=O, S(=O)2; R1 is selected from H, C1-C6 alkyl; L is selected from a bond, C1-C6 alkylene, -C(=O)-, -NR e C(=O)-、-C(=O)-N(R e )-, -OC(=O)-, -C(=O)O-; R e Selected from hydrogen or C1-C6 alkyl; Ar is selected from pyrrolyl, furanyl, thienyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, phenyl; the above groups are optionally substituted by one or more groups selected from the following: halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy; Provided that: when X is S=O, L is not a bond.

6. The compound according to any one of claims 1 to 4, which is a compound represented by formula (III): or its stereoisomers, tautomers, solvates, prodrugs, isotope-labeled substances, and pharmaceutically acceptable salts thereof, wherein: X is selected from S=O, S(=O)2; R1 is selected from H, C1-C6 alkyl; L is selected from a bond, C1-C6 alkylene, -C(=O)-, -NR e C(=O)-、-C(=O)-N(R e )-, -OC(=O)-, -C(=O)O-; R e Selected from hydrogen or C1-C6 alkyl; Ar is selected from pyrrolyl, furanyl, thienyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, phenyl; the above groups are optionally substituted by one or more groups selected from the following: halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy.

7. The compound according to any one of claims 1 to 4, which is a compound represented by formula (IV): or its stereoisomers, tautomers, solvates, prodrugs, isotope-labeled substances, and pharmaceutically acceptable salts thereof, wherein: X is selected from S=O, S(=O)2; R1 is selected from H, C1-C6 alkyl; L is selected from a bond, C1-C6 alkylene, -C(=O)-, -NR e C(=O)-、-C(=O)-N(R e )-, -OC(=O)-, -C(=O)O-; R e Selected from hydrogen or C1-C6 alkyl; Ar is selected from pyrrolyl, furanyl, thienyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, phenyl; the above groups are optionally substituted by one or more groups selected from the following: halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy; Provided that: when X is S=O, L is not a bond.

8. The compound according to any one of claims 1 to 4, which is a compound represented by formula (V): or its stereoisomers, tautomers, solvates, prodrugs, isotope-labeled substances, and pharmaceutically acceptable salts thereof, wherein: X is selected from S=O, S(=O)2; R1 is selected from H, C1-C6 alkyl; R4 is selected from carboxyl, -SO3H, -PO4H, tetrazolyl, triazolyl, -C(=O)OC1-C6 alkyl, - C(=O)NR c R d ; R a , R b are each independently selected from hydrogen, C1-C3 alkyl, halogen, or R a , R b Together with the carbon atom to which it is connected, it forms a cyclopropane group or a cyclobutane group; R c , R d are each independently selected from hydrogen, C1-C3 alkyl, or R c , R d Together with the nitrogen atom to which it is attached, it forms a 3- to 6-membered heterocyclic group; R5 is selected from halogen, hydroxy, amino, thiol, C1-C6 alkyl, C1-C6 alkoxy; Het is selected from 5- or 6-membered nitrogen aromatic group; L is selected from a bond, C1-C6 alkylene, -C(=O)-, -NR e C(=O)-、-C(=O)-N(R e )-, -OC(=O)-, -C(=O)O-; R e Selected from hydrogen or C1-C6 alkyl; Ar is selected from pyrrolyl, furanyl, thienyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, phenyl; the above groups are optionally substituted by one or more groups selected from the following: halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy; p is selected from 1, 2 or 3; q is selected from 1, 2 or 3.

9. The compound according to claim 8 or its stereoisomers, tautomers, solvates, prodrugs, isotope-labeled substances, and pharmaceutically acceptable salts thereof, wherein: X is selected from S=O, S(=O)2; R1 is selected from H, C1-C6 alkyl; R4 is selected from carboxyl, -SO3H, -PO4H, tetrazolyl, triazolyl, -C(=O)OC1-C6 alkyl; R a , R b are each independently selected from hydrogen, halogen, or R a , R b Together with the carbon atom to which it is attached, it forms a cyclopropane group or a cyclobutane group; R5 is selected from halogen, hydroxy, amino, thiol, C1-C6 alkyl, C1-C6 alkoxy; Het is selected from triazole; L is selected from a bond, C1-C6 alkylene, -C(=O)-, -NR e C(=O)-、-C(=O)-N(R e )-, -OC(=O)-, -C(=O)O-; R e Selected from hydrogen or C1-C6 alkyl; Ar is selected from pyridyl, pyrimidinyl, phenyl; the above groups are optionally substituted by one or more groups selected from the following: halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy; p is selected from 1; q is selected from 1.

10. The compound according to claim 1, selected from: or its stereoisomers, tautomers, solvates, prodrugs, isotope-labeled substances and pharmaceutically acceptable salts thereof.

11. A pharmaceutical composition comprising the compound according to any one of claims 1 to 10 or its stereoisomers, tautomers, solvates, prodrugs, isotope-labeled substances and pharmaceutically acceptable salts thereof, and a pharmaceutically acceptable carrier.

12. Use of the compound according to any one of claims 1 to 10 or its stereoisomers, tautomers, solvates, prodrugs, isotope-labeled substances and pharmaceutically acceptable salts thereof and the pharmaceutical composition according to claim 11 in the preparation of a medicament for preventing and / or treating a disease mediated by phosphodiesterase 4B.

13. The use according to claim 12, wherein the disease is an inflammatory disease.

14. The use according to claim 12, wherein the inflammatory disease mediated by phosphodiesterase 4B includes but is not limited to atopic dermatitis, rheumatoid arthritis, asthma, chronic obstructive pulmonary disease (COPD), idiopathic pulmonary fibrosis, interstitial lung disease, chronic sinusitis, allergic rhinitis, allergic dermatitis, contact dermatitis, psoriasis, systemic lupus erythematosus, ulcerative colitis, Crohn's disease, depression, bipolar depression, mania, anxiety, schizophrenia, Alzheimer's disease, stroke, chronic pain, liver fibrosis, kidney fibrosis, and nephritis.

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