Pyrimidine-4(3H)-one heterocyclic compound, preparation method thereof and application thereof in medicine

Abstract

The present invention relates to pyrimidine-4(3 H )-ketone heterocyclic compounds, preparation methods thereof and their applications in medicine. Specifically, the present invention relates to a compound represented by general formula (I) and a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing the compound or its pharmaceutically acceptable salt, a method for using the compound or its pharmaceutically acceptable salt to treat and / or prevent related diseases mediated by SHP2, especially cancer, and a method for preparing the compound or its pharmaceutically acceptable salt. The present invention also relates to the use of the compound or its pharmaceutically acceptable salt or a pharmaceutical composition containing the compound or its pharmaceutically acceptable salt in the preparation of a drug for treating and / or preventing related diseases mediated by SHP2. The substituents of the general formula (I) are the same as those defined in the specification.

Description

Technical Field

[0001] The present invention relates to a novel pyrimidine-4 (3 H )-ketone heterocyclic compounds or pharmaceutically acceptable salts thereof, pharmaceutical compositions containing the compounds or pharmaceutically acceptable salts thereof, methods for preparing the compounds or pharmaceutically acceptable salts thereof, and uses of the compounds or pharmaceutically acceptable salts or pharmaceutical compositions containing the compounds or pharmaceutically acceptable salts thereof in preparing drugs for treating and / or preventing SHP2-mediated related disorders, particularly cancers, and methods for using the same. Background Art

[0002] SHP2 (Src Homology 2 domain-containing Phosphatase 2) is a non-receptor protein tyrosine phosphatase encoded by the PTPN11 (Protein Tyrosine Phosphatase Nonreceptor type 11) gene. SHP2 contains two SH2 domains (Src Homology domain), a PTP domain (ProteinTyrosine Phosphatase domain) and a C-terminal tail. Under normal conditions, SHP2 is in an autoinhibitory conformation, and its N-SH2 binds to PTP, blocking the substrate channel of the PTP catalytic site, thereby inhibiting the activity of PTP. When SH2 binds to a diphosphotyrosine peptide (such as IRS-1), the autoinhibitory interaction of SH2-PTP is released, exposing the PTP catalytic site, making SHP2 in an activated state and catalyzing the dephosphorylation reaction of tyrosine.

[0003] SHP2 is widely expressed. As an oncogene, it mediates the activation of multiple oncogene signal transduction pathways, such as RAS-ERK, PI3K-AKT, and JAK-STAT pathways, promoting the survival and proliferation of cancer cells. SHP2 can bind to and dephosphorylate RAS, increase RAS-RAF association, and activate downstream cell proliferation signals. SHP2 also mediates compensatory activation pathways after kinases such as MEK are inhibited, thereby promoting tumor resistance (Ruess DA, et al., Nat. Med. 2018, 24, 954-960). Therefore, the activation of SHP2 is closely related to the occurrence of various diseases, such as leukemia, melanoma, breast cancer, lung cancer, colon cancer, neuroblastoma, hepatocellular carcinoma, etc.

[0004] SHP2 also plays an important role in the immune checkpoint pathway of PD-1 and BTLA (B- and T-Lymphocyte Attenuator), not only inhibiting T-cell activation but also promoting T-cell dysfunction (Li J, et al., Cancer Res. 2015, 75, 508-518).

[0005] Therefore, SHP2 has attracted a lot of attention as an anti-tumor target. However, due to the high protein sequence homology of the PTP catalytic site and the high hydrophilicity of the PTP catalytic pocket, the SHP2 catalytic site inhibitors have poor selectivity, poor cell permeability and low bioavailability. The discovery of Novartis' allosteric inhibitor SHP099 (Chen Y, et al., Nature 2016, 535, 148-52) provides a new channel for the development of highly specific oral SHP2 inhibitors. At present, several companies have disclosed patent applications for SHP2 inhibitors, including WO2015107495, WO2016203405, WO2018057884, WO2018013597, WO2017211303, etc. The present invention designs a compound having a structure shown in general formula (I), and finds that compounds having such a structure exhibit excellent SHP2 activity inhibition effects. Summary of the invention

[0006] The present invention provides a compound represented by the general formula (I) as a SHP2 inhibitor or its prodrug, stable isotope derivative, pharmaceutically acceptable salt, isomer and mixture thereof:

[0007]

[0008] in:

[0009] Ring A is a benzene ring or a 6-membered heteroaromatic ring, and ring B is a 5-membered heteroaromatic ring fused to ring A, wherein the benzene ring and the heteroaromatic ring are optionally substituted by one or more selected from D, halogen, cyano, oxo, C 1-6 Alkyl, C 3-6 Cycloalkyl, 4-7 membered heterocyclyl, 5-6 membered heteroaryl, -OR a 、-NR a R b 、-C(O)R a 、-C(O)NR a R b 、-S(O)2R a 、-S(O)2NR a R b 、-NR a S(O)2R bor -P(O)(CH3)2, wherein the alkyl, cycloalkyl, heterocyclic and heteroaryl groups are optionally substituted with one or more selected from D, halogen, cyano, oxo, C 1-6 Alkyl, C 3-6 Cycloalkyl, 4-7 membered heterocyclic group, -OR a 、-NR a R b 、-C(O)R a or -C(O)NR a R b substituted by a substituent;

[0010] R 1 H, D, cyano, C 1-2 Alkyl, cyclopropyl, -OR a 、-NR a R b or -C(O)NR a R b , one or more hydrogen atoms of the alkyl and cyclopropyl groups are optionally substituted with one or more D atoms or fluorine atoms;

[0011] R 2 H, C 1-2 Alkyl or cyclopropyl, wherein one or more hydrogen atoms of the alkyl and cyclopropyl groups are optionally substituted by one or more atoms selected from D, fluorine or hydroxyl groups;

[0012] R 4a and R 4b are each independently selected from H, D, halogen, cyano, -OR a 、-NR a R b 、-C(O)R a 、-C(O)NR a R b , C 1-6 Alkyl, C 3-6 Cycloalkyl, 4-7 membered heterocyclyl or 5-6 membered heteroaryl, but R 4a and R 4b Cannot be cyano, -OR at the same time a or -NR a R b The alkyl, cycloalkyl, heterocyclic and heteroaryl groups are optionally substituted by one or more selected from D, halogen, cyano, oxo, C 1-6 Alkyl, C 3-6 Cycloalkyl, 4-7 membered heterocyclyl, 5-6 membered heteroaryl, -OR a 、-NR a R b 、-C(O)R a or -C(O)NR a R bSubstituents; R 4a and R 4b Optional carbon atoms connected to form a C 3-7 Carbocyclic or 4-8 membered heterocyclic ring;

[0013] R 5a , R 5b , R 6a and R 6b Each is independently selected from H, D, fluoro or methyl;

[0014] R a and R b Each independently selected from H, C 1-6 Alkyl, C 3-6 Cycloalkyl or 4-7 membered heterocyclyl, wherein the alkyl, cycloalkyl and heterocyclyl are optionally substituted by one or more substituents selected from D, fluoro, cyano, oxo, hydroxyl, -OCH3 or -NH2.

[0015] One embodiment of the present invention relates to a compound represented by the above general formula (I) or a pharmaceutically acceptable salt, prodrug, stable isotope derivative, isomer and a mixture thereof, which is a compound represented by the general formula (II) or a pharmaceutically acceptable salt, prodrug, stable isotope derivative, isomer and a mixture thereof:

[0016]

[0017] in:

[0018] Ring A is a benzene ring or a 6-membered heteroaromatic ring, and ring B is a 5-membered heteroaromatic ring fused to ring A, wherein the benzene ring and the heteroaromatic ring are optionally substituted by one or more selected from D, halogen, cyano, oxo, C 1-6 Alkyl, C 3-6 Cycloalkyl, 4-7 membered heterocyclyl, 5-6 membered heteroaryl, -OR a 、-NR a R b 、-C(O)R a 、-C(O)NR a R b 、-S(O)2R a 、-S(O)2NR a R b or -NR a S(O)2R b wherein the alkyl, cycloalkyl, heterocyclic and heteroaryl groups are optionally substituted with one or more substituents selected from D, halogen, cyano, oxo, C 1-6 Alkyl, C 3-6 Cycloalkyl, 4-7 membered heterocyclic group, -OR a 、-NR a R b、-C(O)R a or -C(O)NR a R b substituted by a substituent;

[0019] R 1 H, D, cyano, C 1-2 Alkyl, -OR a or -NR a R b , one or more hydrogen atoms of the alkyl group are optionally substituted by one or more D atoms or fluorine atoms;

[0020] R 2 H or C 1-2 Alkyl, wherein one or more hydrogen atoms of the alkyl group are optionally substituted by one or more D or fluorine atoms;

[0021] R 4a and R 4b are each independently selected from H, D, halogen, cyano, -OR a 、-NR a R b 、-C(O)NR a R b , C 1-6 Alkyl, C 3-6 Cycloalkyl, 4-7 membered heterocyclyl or 5-6 membered heteroaryl, but R 4a and R 4b Cannot be cyano, -OR at the same time a or -NR a R b The alkyl, cycloalkyl, heterocyclic and heteroaryl groups are optionally substituted by one or more selected from D, halogen, cyano, oxo, C 1-6 Alkyl, C 3-6 Cycloalkyl, 4-7 membered heterocyclyl, 5-6 membered heteroaryl, -OR a 、-NR a R b 、-C(O)R a or -C(O)NR a R b Substituents; R 4a and R 4b Optional carbon atoms connected to form a C 3-7 Carbocyclic or 4-7 membered heterocyclic ring;

[0022] R a and R b Each independently selected from H, C 1-6 Alkyl, C 3-6Cycloalkyl or 4-7 membered heterocyclyl, wherein the alkyl, cycloalkyl and heterocyclyl are optionally substituted by one or more substituents selected from D, fluoro, cyano, oxo, hydroxyl, -OCH3 or -NH2.

[0023] Another embodiment of the present invention relates to the compound represented by the above general formula (I) or its pharmaceutically acceptable salts, prodrugs, stable isotope derivatives, isomers and mixtures thereof, which is a compound represented by general formula (III) or its pharmaceutically acceptable salts, prodrugs, stable isotope derivatives, isomers and mixtures thereof:

[0024]

[0025] in:

[0026] Ring A is a benzene ring or a 6-membered heteroaromatic ring, and ring B is a 5-membered heteroaromatic ring fused to ring A, wherein the benzene ring and the heteroaromatic ring are optionally substituted by one or more selected from D, halogen, cyano, oxo, C 1-6 Alkyl, C 3-6 Cycloalkyl, 4-7 membered heterocyclyl, 5-6 membered heteroaryl, -OR a 、-NR a R b 、-C(O)R a or -C(O)NR a R b wherein the alkyl, cycloalkyl, heterocyclic and heteroaryl groups are optionally substituted with one or more substituents selected from D, halogen, cyano, oxo, C 1-2 Alkyl, -OR a or -NR a R b substituted by a substituent;

[0027] X is -O- or -CR 8a R 8b -;

[0028] R 1 H, D, cyano, C 1-2 Alkyl or -NR a R b , one or more hydrogen atoms of the alkyl group are optionally substituted by one or more D atoms or fluorine atoms;

[0029] R 2 H or C 1-2 Alkyl, wherein one or more hydrogen atoms of the alkyl group are optionally substituted by one or more D or fluorine atoms;

[0030] R 7a , R 7b , R 8a , R 8b , R 9a and R 9bare each independently selected from H, D, halogen, cyano, C 1-2 Alkyl or -OR c , one or more hydrogen atoms of the alkyl group are optionally substituted by one or more atoms selected from D, fluorine or hydroxyl groups, but R 7a and R 7b , R 8a and R 8b , R 9a and R 9b Optional cannot be -OR at the same time c ;

[0031] R a and R b Each independently selected from H, C 1-6 Alkyl, C 3-6 Cycloalkyl or 4-7 membered heterocyclyl, wherein the alkyl, cycloalkyl and heterocyclyl are optionally substituted by one or more substituents selected from D, fluoro, cyano, oxo, hydroxyl, -OCH3 or -NH2;

[0032] R c H or C 1-2 Alkyl, wherein one or more hydrogen atoms of the alkyl group are optionally substituted by one or more D atoms or fluorine atoms.

[0033] Another embodiment of the present invention relates to the compound represented by the above general formula (I) or its pharmaceutically acceptable salts, prodrugs, stable isotope derivatives, isomers and mixtures thereof, which is a compound represented by general formula (IV) or its pharmaceutically acceptable salts, prodrugs, stable isotope derivatives, isomers and mixtures thereof:

[0034]

[0035] in:

[0036] Ring A is a benzene ring or a 6-membered heteroaromatic ring, and ring B is a 5-membered heteroaromatic ring fused to ring A, wherein the benzene ring and the heteroaromatic ring are optionally substituted by one or more selected from D, halogen, cyano, oxo, C 1-2 Alkyl, -OR a or -NR a R b wherein one or more hydrogen atoms of the alkyl group are optionally substituted with one or more D atoms or fluorine atoms;

[0037] X is -O- or -CH2-;

[0038] R 1 Independently selected from H, D, C 1-2 Alkyl or -NR a R b , one or more hydrogen atoms of the alkyl group are optionally substituted by one or more D atoms or fluorine atoms;

[0039] R 2 H or C 1-2 Alkyl, wherein one or more hydrogen atoms of the alkyl group are optionally substituted by one or more D or fluorine atoms;

[0040] R 7 H, D or C 1-2 Alkyl, wherein one or more hydrogen atoms of the alkyl group are optionally substituted by one or more D or fluorine atoms;

[0041] R a and R b Each independently is H or C 1-2 Alkyl, wherein one or more hydrogen atoms of the alkyl group are optionally substituted by one or more D atoms or fluorine atoms.

[0042] The present invention further relates to a compound represented by the above general formula (I), wherein the compound is selected from:

[0043]

[0044]

[0045]

[0046] or a pharmaceutically acceptable salt, prodrug, stable isotope derivative, isomer or a mixture thereof.

[0047] The compounds of the present invention can effectively inhibit the activity of SHP2, and preferably have an IC 50 The compounds of the present invention have a significant inhibitory effect on the proliferation of NCI-H358 cells, preferably with an IC 50 Less than 1000 nM.

[0048] Another aspect of the present invention relates to a pharmaceutical composition comprising a compound represented by general formula (I) or a pharmaceutically acceptable salt, prodrug, stable isotope derivative, isomer and mixture thereof and a pharmaceutically acceptable carrier and excipient.

[0049] The present invention further relates to a pharmaceutical composition, comprising a compound represented by general formula (I) or a pharmaceutically acceptable salt, prodrug, stable isotope derivative, isomer and a mixture thereof and at least one additional drug, wherein the at least one additional drug includes but is not limited to chemotherapeutic drugs, targeted drugs, DNA synthesis inhibitors, antibody drugs, antibody drug conjugates, anti-tumor drugs, immunosuppressants, etc.

[0050] Another aspect of the present invention also relates to the use of the compound represented by general formula (I) or its pharmaceutically acceptable salts, prodrugs, stable isotope derivatives, isomers and mixtures thereof or the pharmaceutical composition in the preparation of drugs, wherein the drugs are used to treat or prevent SHP2-mediated related diseases, wherein the diseases include but are not limited to leukemia, Noonan syndrome, leopard spot syndrome, neuroblastoma, lung cancer, breast cancer, colon cancer, esophageal cancer, gastric cancer, head and neck cancer, etc.

[0051] According to the present invention, the drug can be in any pharmaceutical dosage form, including but not limited to tablets, capsules, solutions, lyophilized preparations, injections, and the like.

[0052] The pharmaceutical preparations of the present invention can be administered in the form of dosage units containing a predetermined amount of active ingredient per dosage unit. Such units may contain, for example, 0.5 mg to 1 g, preferably 1 mg to 700 mg, and particularly preferably 5 mg to 500 mg of the compound of the present invention, depending on the condition to be treated, the method of administration, and the age, weight, and condition of the patient. In addition, pharmaceutical preparations of this type may be prepared using methods known in the pharmaceutical field, such as mixing the active ingredient with one or more auxiliary materials and / or adjuvants.

[0053] The pharmaceutical formulations of the invention may be adapted for administration by any desired suitable method, for example by oral (including buccal or sublingual), rectal, nasal, topical (including buccal, sublingual or transdermal), vaginal or parenteral (including subcutaneous, intramuscular, intravenous or intradermal) administration. DETAILED DESCRIPTION

[0054] Unless stated otherwise to the contrary, the following terms used in the specification and claims have the following meanings.

[0055] “C x-y " represents the range of carbon number, where x and y are both integers, e.g. 3-8 Cycloalkyl means a cycloalkyl group having 3 to 8 carbon atoms, i.e. a cycloalkyl group having 3, 4, 5, 6, 7 or 8 carbon atoms. 3-8 " also includes any sub-ranges therein, such as C 3-7 , C 3-6 , C 4-7 , C 4-6 , C 5-6 wait.

[0056] "Alkyl" refers to a saturated straight or branched hydrocarbon group containing 1 to 20 carbon atoms, such as 1 to 8 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms. Non-limiting examples of alkyl include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, etc.

[0057] "Cycloalkyl or carbocycle" refers to a saturated cyclic hydrocarbon substituent containing 3 to 14 carbon ring atoms. Cycloalkyl can be a single carbocycle, typically containing 3 to 8, 3 to 7, or 3 to 6 carbon ring atoms. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, etc. Cycloalkyl can also be a double or tricyclic ring fused, bridged, or spiro-bound together, such as decahydronaphthyl, bicyclo[2.2.2]octane, spiro[3.3]heptane, etc.

[0058] "Heterocyclyl or heterocycle" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic group comprising 3 to 20 ring atoms, for example 3 to 14, 3 to 12, 3 to 10, 3 to 8, 3 to 6 or 5 to 6 ring atoms, wherein one or more ring atoms are selected from nitrogen, oxygen or S(O) m (wherein m is an integer 0 to 2), but does not include the ring part of -OO-, -OS- or -SS-, and the remaining ring atoms are carbon. Preferably 3 to 12 ring atoms are included, more preferably 3 to 10 ring atoms, more preferably 4 to 7 ring atoms, more preferably 4 to 6 ring atoms, most preferably 5 or 6 ring atoms, wherein 1 to 4 are heteroatoms, more preferably 1 to 3 are heteroatoms, and most preferably 1 to 2 are heteroatoms. The non-limiting examples of monocyclic heterocyclic radicals include pyrrolidinyl, oxadiazine, piperidinyl, piperazinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiopyranyl, morpholinyl, thiomorpholinyl, homopiperazinyl, azetidinyl etc. The polycyclic heterocyclic group includes fused, bridged or spiro polycyclic heterocyclic groups, such as octahydrocyclopenta[c]pyrrole, octahydropyrrolo[1,2-a]pyrazine, 3,8-diazabicyclo[3.2.1]octane, 5-azaspiro[2.4]heptane, 2-oxa-7-azaspiro[3.5]nonane and the like.

[0059] "Aryl or aromatic ring" refers to an aromatic monocyclic or fused polycyclic group containing 6 to 14 carbon atoms, preferably 6 to 10 members, such as phenyl and naphthyl, most preferably phenyl. The aryl ring may be fused to a heteroaryl, heterocyclyl or cycloalkyl ring, wherein the ring connected to the parent structure is an aryl ring, non-limiting examples include:

[0060]

[0061] wait.

[0062] "Heteroaryl or heteroaromatic ring" refers to a heteroaromatic system containing 5 to 14 ring atoms, wherein 1 to 4 ring atoms are selected from heteroatoms including oxygen, sulfur and nitrogen. Heteroaryl is preferably 5 to 10 yuan, and more preferably heteroaryl is 5 or 6 yuan, such as furanyl, thienyl, pyridyl, pyrrolyl, pyrimidinyl, pyrazinyl, pyrazolyl, imidazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, quinolyl, isoquinolyl, indolyl, isoindolyl, etc. The heteroaryl ring can be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring, non-limiting examples include:

[0063]

[0064] wait.

[0065] "Halogen" refers to fluorine, chlorine, bromine or iodine.

[0066] "Cyano" refers to -CN.

[0067] "Oxo" refers to =0.

[0068] "Optional" means that the subsequently described event or circumstance may but need not occur, and the description includes the case where the event or circumstance occurs or does not occur. For example, "a heterocyclic group optionally substituted with an alkyl group" means that the alkyl group may but need not be present, and the description includes the case where the heterocyclic group is substituted with an alkyl group and the case where the heterocyclic group is not substituted with an alkyl group.

[0069] "Substitution" means that one or more hydrogen atoms, preferably 5, more preferably 1 to 3 hydrogen atoms in a group are replaced independently of each other by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) possible or impossible substitutions without undue effort. For example, amino or hydroxyl groups with free hydrogen may be unstable when combined with carbon atoms with unsaturated (such as olefinic) bonds. The substituents include, but are not limited to, halogen, cyano, nitro, oxo, -SF5, C 1-4 Alkyl, C 3-7 Cycloalkyl, 4-7 membered heterocyclic group, phenyl, 5-6 membered heteroaryl, etc.

[0070] "Isomers" refer to compounds that have the same molecular formula but differ in the nature or order of bonding of their atoms or in the arrangement of their atoms in space. Isomers that differ in the arrangement of their atoms in space are called "stereoisomers". Stereoisomers include optical isomers, geometric isomers and conformational isomers.

[0071] The compounds of the present invention may exist in the form of optical isomers. Optical isomers include enantiomers and diastereomers. Enantiomers refer to two stereoisomers that are mirror images of each other but cannot overlap each other. A racemic mixture or racemate refers to a mixture of equal amounts of left-handed and right-handed enantiomers of a chiral molecule. Diastereomers refer to two stereoisomers that are not mirror images of each other and cannot overlap. When an optical isomer is a single isomer and its absolute configuration is determined, it is an absolute configuration of "R" or "S" according to the configuration of the substituents on the chiral carbon atom; when the absolute configuration of an optical isomer is undetermined, it is (+) or (-) according to the measured optical rotation value. Methods for preparing and separating optical isomers are known in the art.

[0072] The compounds of the present invention may also exist as geometric isomers. The present invention contemplates various geometric isomers and mixtures thereof generated by the distribution of substituents around carbon-carbon double bonds, carbon-nitrogen double bonds, cycloalkyl or heterocyclic groups. Substituents around carbon-carbon double bonds or carbon-nitrogen bonds are designated as Z or E configurations, and substituents around cycloalkyl or heterocyclic rings are designated as cis or trans configurations.

[0073] The compounds of the present invention may also exhibit tautomerism, such as keto-enol tautomerism.

[0074] It should be understood that the present invention includes any tautomeric or stereoisomeric forms and mixtures thereof and is not limited solely to any one tautomeric or stereoisomeric form used in the naming of the compound or chemical formula.

[0075] "Isotopes" refers to all isotopes of atoms present in the compounds of the present invention. Isotopes include those atoms having the same atomic number but different mass numbers. Examples of isotopes suitable for incorporation into the compounds of the present invention are hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine, such as but not limited to 2 H (D), 3 H. 13 C. 14 C. 15 N. 17 O. 18 O. 31 P. 32 P. 35 S. 18 F and 36Cl. Isotope-labeled compounds of the present invention can generally be prepared by conventional techniques known to those skilled in the art or by methods similar to those described in the accompanying Examples using appropriate isotope-labeled reagents in place of non-isotope-labeled reagents. Such compounds have various potential uses, for example as standards and reagents in determining biological activity. In the case of stable isotopes, such compounds have the potential to favorably alter biological, pharmacological or pharmacokinetic properties. Deuterium (D) is a preferred isotope of the present invention, such as hydrogen in a methyl, methylene or methine group can be replaced by deuterium.

[0076] The compounds of the present invention can be given in the form of prodrugs. "Prodrug" refers to derivatives of the bioactive compounds of the present invention that are converted into, for example, by oxidation, reduction, hydrolysis, etc. (they are each carried out using enzymes or without enzyme participation) under physiological conditions in vivo. Examples of prodrugs are the following compounds: wherein the amino group in the compounds of the present invention is acylated, alkylated or phosphorylated, such as eicosanoylamino, alanylamino, pivaloyloxymethylamino, or wherein the hydroxyl group is acylated, alkylated, phosphorylated or converted into borate, such as acetoxy, palmitoyloxy, pivaloyloxy, succinyloxy, fumaryloxy, alanyloxy, or wherein the carboxyl group is esterified or amidated, or wherein the sulfhydryl group is selectively delivered to the target and / or to the carrier molecule of the cytosol of the cell, such as a peptide to form a disulfide bridge. These compounds can be prepared by the compounds of the present invention according to known methods.

[0077] "Pharmaceutically acceptable salts" or "pharmaceutically acceptable salts" refer to salts made from pharmaceutically acceptable bases or acids, including inorganic bases or acids and organic bases or acids. In the case where the compounds of the present invention contain one or more acidic or basic groups, the present invention also includes their corresponding pharmaceutically acceptable salts. Therefore, the compounds of the present invention containing acidic groups can exist in salt form and can be used according to the present invention, for example as alkali metal salts, alkaline earth metal salts or as ammonium salts. More specific examples of such salts include sodium salts, potassium salts, calcium salts, magnesium salts or salts with ammonia or organic amines, such as ethylamine, ethanolamine, triethanolamine or amino acids. The compounds of the present invention containing basic groups can exist in salt form and can be used according to the present invention in the form of addition salts thereof with inorganic or organic acids. The example of suitable acid comprises hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, nitric acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenedisulfonic acid, oxalic acid, acetic acid, tartaric acid, lactic acid, salicylic acid, benzoic acid, formic acid, propionic acid, pivalic acid, malonic acid, succinic acid, pimelic acid, fumaric acid, maleic acid, malic acid, sulfamic acid, phenylpropionic acid, gluconic acid, ascorbic acid, isonicotinic acid, citric acid, adipic acid and other acid well known to persons skilled in the art. If the compound of the present invention contains acidic and basic groups in the molecule simultaneously, the present invention also comprises inner salt or betaine except the salt form mentioned. Each salt can be obtained by conventional methods well known to persons skilled in the art, for example by making these contact with organic or inorganic acid or base in solvent or dispersant or by with other salt anion exchange or cation exchange.

[0078] "Pharmaceutical composition" refers to a composition containing one or more compounds described herein or their pharmaceutically acceptable salts, prodrugs, stable isotope derivatives, isomers and mixtures thereof, as well as other components such as pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitate the absorption of the active ingredient, and thus exert biological activity.

[0079] Therefore, when referring to "compound", "compound of the present invention" or "compound described in the present invention" in this application, all forms of the compound, such as pharmaceutically acceptable salts, prodrugs, stable isotope derivatives, isomers and mixtures thereof, are included.

[0080] As used herein, the term "therapeutically effective amount" refers to an amount of the compound of the present invention that is effective in inhibiting the function of SHP2 and / or treating or preventing the disease.

[0081] As used herein, the term "patient" refers to a mammal, especially a human.

[0082] Synthesis method

[0083] The present invention also provides a method for preparing the compound. The preparation of the compound described in the general formula (I) of the present invention can be completed by the following exemplary methods and examples, but these methods and examples should not be considered to limit the scope of the present invention in any way. The compound described in the present invention can also be synthesized by synthetic techniques known to those skilled in the art, or a combination of methods known in the art and the method described in the present invention. The product obtained from each step of the reaction is obtained by separation techniques known in the art, including but not limited to extraction, filtration, distillation, crystallization, chromatographic separation, etc. The starting materials and chemical reagents required for the synthesis can be conventionally synthesized or purchased according to the literature (which can be queried from SciFinder).

[0084] The heterocyclic compound of the general formula (I) of the present invention can be synthesized according to the route described in method A: 1) A1 and the NH in the piperidine compound A2 are subjected to a substitution reaction under base catalysis or a condensation reaction using a condensation agent to obtain an intermediate A3; 2) A3 is then subjected to a Buchwald coupling reaction with the thiol group on the 5,6-fused heteroaromatic ring to generate a product A4.

[0085] Method A:

[0086]

[0087] The heterocyclic compound of the general formula (I) of the present invention can be synthesized according to the route described in method B: 1) A1 and A2a are subjected to a substitution reaction under base catalysis or a condensation reaction using a condensation agent to obtain an intermediate A3a; 2) the keto group in A3a and ( R )-2-methylpropane-2-sulfenamide undergoes reductive amination to generate A5a; 3) A5a then reacts with the thiol group on the 5,6-fused heteroaromatic ring via Buchwald coupling to generate A6a; 4) A6a is deprotected in an acidic environment to obtain the product A4a.

[0088] Method B:

[0089] Example

[0090] The starting materials of the present invention can be synthesized by methods known in the art, or can be purchased from chemical companies such as ABCR MBH & Co. KG, Acros Organics, Aldrich Chemical Company, AccelaChemBio Inc., and Beijing Coupling Chemicals.

[0091] The structure of the compound was determined by nuclear magnetic resonance (NMR) or mass spectrometry (MS). NMR was measured using a Bruker ASCEND-400 NMR spectrometer, and the solvent was deuterated dimethyl sulfoxide (DMSO- d6 ), deuterated chloroform (CDC13), deuterated methanol (CD3OD), etc. The internal standard is tetramethylsilane (TMS), and the chemical shift is 10 -6 The units are given as (ppm). MS was measured using an Agilent SQD (ESI) mass spectrometer (Agilent 6120).

[0092] HPLC was performed using an Agilent 1260 DAD high pressure liquid chromatograph (Poroshell120 EC-C18, 50×3.0 mm, 2.7 μm column) or a Waters Arc high pressure liquid chromatograph (Sunfire C18, 150×4.6 mm, 5 μm column).

[0093] Unless otherwise specified in the examples, the reaction temperature is room temperature (20-30°C).

[0094] Unless otherwise specified in the examples, the reactions were carried out under an argon atmosphere or a nitrogen atmosphere. The argon atmosphere or the nitrogen atmosphere means that the reaction flask is connected to an argon or nitrogen balloon with a volume of about 1 L.

[0095] Hydrogen atmosphere means that the reaction bottle is evacuated and then filled with hydrogen (repeated 3 times), and then connected to a hydrogen balloon with a volume of about 1 L.

[0096] The microwave reaction was carried out using a CEM Discover-SP microwave reactor.

[0097] The reaction progress in the examples was monitored using an Agilent LC-MS / MS chromatograph (1260 / 6120), or thin layer chromatography (TLC), with the thickness of the silica gel plate being 0.15-0.2 mm (Qingdao Ocean GF254).

[0098] The compounds were purified by column chromatography or thin layer chromatography, wherein the column chromatography used 200-300 mesh silica gel from Qingdao Ocean, and the thin layer chromatography used GF254 silica gel plate with a thickness of 0.4-0.5 mm from Qingdao Ocean.

[0099] The developing solvent system for column chromatography or thin layer chromatography is usually a) dichloromethane and methanol system, b) petroleum ether and ethyl acetate system, or as shown in the examples. The volume ratio of the solvent is adjusted according to the polarity of the compound, and a small amount of triethylamine or other acidic or basic reagents can be added to further adjust.

[0100] The compounds were purified using a Waters mass spectrometry-guided automated preparation system (mass spectrometry detector: SQD2), and the compounds were eluted with a reversed phase high pressure column (XBridge-C18, 19×150 mm, 5 μm) at a flow rate of 20 mL / min using an appropriate acetonitrile / water (containing 0.1% trifluoroacetic acid or formic acid, or 0.05% ammonia) gradient according to the polarity of the compounds. Some examples can be purified using an automated preparation system by adding 1 N dilute hydrochloric acid, and then removing the solvent under reduced pressure to obtain a hydrochloride.

[0101] The abbreviation DMF refers to N,N-dimethylformamide.

[0102] The abbreviation DIPEA refers to N,N-diisopropylethylamine.

[0103] The abbreviation DBU refers to 1,8-diazabicyclo[5.4.0]undec-7-ene.

[0104] The abbreviation NBS refers to N-bromosuccinimide.

[0105] The abbreviation NIS refers to N-iodosuccinimide.

[0106] The abbreviation XantPhos refers to 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene.

[0107] The abbreviation Pd2(dba)3 refers to tris(dibenzylideneacetone)dipalladium.

[0108] Carter condensation agent refers to benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate.

[0109] Example 1

[0110] ( R )-5-((1H-pyrrolo[2,3- b ]pyridin-4-yl)thio)-2-(1-amino-8-azaspiro[4.5]decane-8-yl)-3-methylpyrimidine-4(3 H )-Ketohydrochloride

[0111]

[0112]

[0113] first step

[0114] 2-Chloro-5-iodopyrimidine-4(3 H )-Keto (1b)

[0115] 2,4-Dichloro-5-iodopyrimidine 1a (8.0 g, 29.1 mmol) was dissolved in tetrahydrofuran (100 mL), and sodium hydroxide solution (1 N, 45 mL) was added at 0°C. The reaction mixture was warmed to room temperature and stirred for 16 hours. The pH value was adjusted to acidic with citric acid and extracted with ethyl acetate (60 mL×3). The organic phases were combined and washed with saturated brine (30 mL×3), dried over anhydrous sodium sulfate, and desolventized under reduced pressure to obtain the target product 1b (6.7 g, solid) with a yield of 91%.

[0116] MS m / z (ESI): 257 [M+1]

[0117] Step 2

[0118] 2-Chloro-5-iodo-3-methylpyrimidine-4(3 H )-Keto (1c)

[0119] Dissolve 1b (5.6 g, 21.7 mmol) in tetrahydrofuran (100 mL), add iodomethane (3.7 g, 26 mmol) and DIPEA (8.4 g, 65.1 mmol), heat to 60°C, and stir for 16 hours. Pour into water (120 mL) and extract with ethyl acetate (80 mL×3). The organic phases were combined and washed with saturated brine (30 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 3 / 7) to obtain the target product 1c (2.2 g, solid), with a yield of 37%.

[0120] MS m / z (ESI): 271 [M+1]

[0121] Step 3

[0122] 8-(5-iodo-1-methyl-6-carbonyl-1,6-dihydropyrimidin-2-yl)-8-azaspiro[4.5]decan-1-one (1d)

[0123] 8-Azaspiro[4.5]decan-1-one (hydrochloride, 586 mg, 3.1 mmol) and potassium carbonate (1.51 g, 11 mmol) were added to a solution of 1c (1.0 g, 3.65 mmol) in acetonitrile (25 mL). The reaction mixture was heated to 80°C and stirred for 16 hours. After cooling to room temperature, the reaction mixture was poured into water (50 mL) and extracted with ethyl acetate (30 mL×3). The organic phases were combined and washed with saturated brine (20 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 7 / 3) to obtain the target product 1d (1.2 g, solid) with a yield of 85%.

[0124] MS m / z (ESI): 388 [M+1]

[0125] Step 4

[0126] ( R )-N-(( R )-8-(5-iodo-1-methyl-6-carbonyl-1,6-dihydropyrimidin-2-yl)-8-azaspiro[4.5]decane-1-yl)-2-methylpropane-2-sulfinamide (1e)

[0127] To a solution of 1d (1.2 g, 3.1 mmol) in tetrahydrofuran (30 mL) was added ( R )-2-methylpropane-2-sulfenamide (750 mg, 6.2 mmol) and tetraethyl titanate (2.82 g, 12.4 mmol), heated to 90°C, and stirred for 16 hours. Cooled to 0°C, methanol (10 mL) and lithium borohydride tetrahydrofuran solution (2.0 M, 1.5 mL, 3 mmol) were added and stirred for 1 hour. Ammonium chloride solution was added to quench and filtered. The filtrate was extracted with ethyl acetate (25 mL×3), and the organic phases were combined and washed with saturated brine (20 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 2 / 1) to obtain the target product 1e (1.4 g, solid), with a yield of 93%.

[0128] MS m / z (ESI): 493 [M+1]

[0129] Step 5

[0130] 3-((1H-pyrrolo[2,3- b ]ethyl pyridin-4-yl)thio)propanoate (1 g)

[0131] To 4-bromo-1H-pyrrolo[2,3- b ] Ethyl 3-mercaptopropionate (408 mg, 3.04 mmol), DIPEA (588 mg, 4.56 mmol), Pd2(dba)3 (140 mg, 0.152 mmol) and XantPhos (132 mg, 0.228 mmol) were added to a solution of pyridine 1f (300 mg, 1.52 mmol) in dioxane (20 mL). The reaction mixture was heated to 100°C under nitrogen and reacted for 2 hours. The temperature was cooled to room temperature and desolventized under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 7 / 3) to obtain the target product 1g (200 mg, oily substance) with a yield of 53%.

[0132] MS m / z (ESI): 251 [M+1]

[0133] Step 6

[0134] 1H-pyrrolo[2,3- b ]Sodium pyridine-4-thiolate (1h)

[0135] To a solution of 1 g (200 mg, 0.8 mmol) in methanol (20 mL), add a methanol solution of sodium methoxide (30%, 158 mg, 0.88 mmol) and stir at room temperature for 2 hours. Remove the solvent under reduced pressure to obtain the target product 1h (100 mg, solid) with a yield of 73%. The product was used directly in the next step without purification.

[0136] MS m / z (ESI): 151 [M+1]

[0137] Step 7

[0138] ( R )-N-(( R )-8-(5-((1H-pyrrolo[2,3- b ]pyridin-4-yl)thio)-1-methyl-6-carbonyl-1,6-dihydropyrimidin-2-yl)-8-azaspiro[4.5]decane-1-yl)-2-methylpropane-2-sulfinamide (1i)

[0139] 1h (55 mg, 0.32 mmol) was dissolved in dioxane (6 mL), and 1e (82 mg, 0.16 mmol), DIPEA (128 mg, 0.99 mmol), Pd2(dba)3 (30 mg, 0.033 mmol) and XantPhos (29 mg, 0.049 mmol) were added. The reaction mixture was heated to 80°C under nitrogen protection and reacted for 2 hours. The mixture was cooled to room temperature and desolventized under reduced pressure. The residue was purified by silica gel column chromatography (methanol / dichloromethane = 1 / 9) to obtain the target product 1i (30 mg, solid) with a yield of 37%.

[0140] MS m / z (ESI): 515 [M+1]

[0141] Step 8

[0142] ( R )-5-((1H-pyrrolo[2,3- b ]pyridin-4-yl)thio)-2-(1-amino-8-azaspiro[4.5]decane-8-yl)-3-methylpyrimidine-4(3 H )-Ketohydrochloride (1)

[0143] 1i (30 mg, 0.058 mmol) was dissolved in methanol (5 mL), and a solution of hydrogen chloride in dioxane (4.0 M, 5 mL) was added, and the mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure, and the residue was purified by reverse phase preparative HPLC to obtain the target product 1 (7.1 mg, solid) with a yield of 31%.

[0144] MS m / z (ESI): 411 [M+1]

[0145] 1 H NMR (400 MHz, DMSO- d 6 ) δ 12.68 (s, 1H), 8.24 – 8.14 (m, 5H), 7.68 (s, 1H), 6.91 – 6.87 (m, 1H), 6.74 (s, 1H), 3.75 – 3.64 (m, 2H), 3.43 (s,3H), 3.19 – 3.08 (m, 3H), 2.08 – 2.05 (m, 1H), 1.92 – 1.64 (m, 7H), 1.54 –1.51 (m, 1H), 1.44 – 1.41 (m, 1H).

[0146] Example 2

[0147] ( R )-2-(1-amino-8-azaspiro[4.5]decane-8-yl)-3-methyl-5-((1-methyl-1H-pyrrolo[2,3- b ]pyridin-4-yl)thio)pyrimidine-4(3 H )-Ketoformate

[0148]

[0149] first step

[0150] 3-((1-methyl-1H-pyrrolo[2,3- b ]4-pyridin-4-yl)thio)propionic acid ethyl ester (2b)

[0151] To 4-bromo-1-methyl-1H-pyrrolo[2,3- b] Pd2(dba)3 (87 mg, 0.095 mmol) was added to a mixture of pyridine 2a (200 mg, 0.95 mmol), ethyl 3-mercaptopropionate (255 mg, 1.9 mmol), DIPEA (368 mg, 2.85 mmol), XantPhos (55 mg, 0.095 mmol) and dioxane (10 mL). The mixture was heated to 100°C under nitrogen protection and reacted for 2 hours. The mixture was cooled to room temperature, water (10 mL) was added, and the mixture was extracted with ethyl acetate (20 mL×3). The organic phase was dried over anhydrous sodium sulfate, the desiccant was removed by filtration, and the solvent was removed under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 3 / 7) to obtain the target product 2b (240 mg, solid) with a yield of 96%.

[0152] MS m / z (ESI): 265 [M+1]

[0153] Step 2

[0154] 1-Methyl-1H-pyrrolo[2,3- b ]Pyridine-4-thiol (2c)

[0155] Potassium tert-butoxide (204 mg, 1.82 mmol) was added to a solution of 2b (240 mg, 0.91 mmol) in tetrahydrofuran (5 mL) and stirred at room temperature for 2 hours. The mixture was acidified to pH 4 with hydrochloric acid (6 N) and extracted with ethyl acetate (20 mL). The organic phase was dried over anhydrous sodium sulfate, filtered to remove the desiccant, and desolventized under reduced pressure to obtain the target product 2c (130 mg, solid) with a yield of 87%.

[0156] MS m / z (ESI): 165 [M+1]

[0157] Step 3

[0158] ( R )-2-methyl-N-(( R )-8-(1-methyl-5-((1-methyl-1H-pyrrolo[2,3- b ]pyridin-4-yl)thio)-6-carbonyl-1,6-dihydropyrimidin-2-yl)-8-azaspiro[4.5]decane-1-yl)propane-2-sulfenamide (2d)

[0159] Pd2(dba)3 (20 mg, 0.022 mmol) was added to a mixture of 2c (50 mg, 0.24 mmol), 1e (110 mg, 0.22 mmol), DIPEA (85 mg, 0.66 mmol), XantPhos (13 mg, 0.022 mmol) and dioxane (5 mL), and the mixture was heated to 100°C under nitrogen protection for 2 hours. The mixture was cooled to room temperature, desolventized under reduced pressure, and the residue was purified by silica gel column chromatography (methanol / dichloromethane = 1 / 9) to obtain the target product 2d (80 mg, solid) with a yield of 68%.

[0160] MS m / z (ESI): 529 [M+1]

[0161] Step 4

[0162] ( R )-2-(1-amino-8-azaspiro[4.5]decane-8-yl)-3-methyl-5-((1-methyl-1H-pyrrolo[2,3- b ]pyridin-4-yl)thio)pyrimidine-4(3 H )-Ketoformate (2)

[0163] 2d (80 mg, 0.14 mmol) was dissolved in methanol (8 mL), and a solution of hydrogen chloride in dioxane (4.0 M, 2 mL) was added, and the mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure, and the residue was purified by reverse phase preparative HPLC to obtain the target product 2 (21.7 mg, solid) with a yield of 34%.

[0164] MS m / z (ESI): 425 [M+1]

[0165] 1 H NMR (400 MHz, DMSO- d 6 ) δ 8.34 (s, 1H), 8.14 (s, 1H), 8.03 (d, J =5.1 Hz, 1H), 7.51 (d, J = 3.5 Hz, 1H), 6.58 (d, J = 5.1 Hz, 1H), 6.44 (d, J =3.5 Hz, 1H), 3.80 (s, 3H), 3.66 – 3.58 (m, 2H), 3.41 (s, 3H), 3.07 (t, J=11.5 Hz, 2H), 2.99 – 2.96 (m, 1H), 2.0 – 1.92 (m, 1H), 1.80 – 1.68 (m, 4H), 1.60 – 1.53 (m, 3H), 1.40 (d, J = 12.9 Hz, 1H), 1.33 (d, J = 13.1 Hz, 1H).

[0166] Example 3

[0167] ( R )-5-((1H-indazol-4-yl)thio)-2-(1-amino-8-azaspiro[4.5]decane-8-yl)-3-methylpyrimidine-4(3 H )-Ketohydrochloride

[0168]

[0169] first step

[0170] Ethyl 3-((1H-indazol-4-yl)thio)propanoate (3b)

[0171] To a solution of 4-bromo-1H-indazole 3a (500 mg, 2.53 mmol) in dioxane (20 mL) were added ethyl 3-mercaptopropionate (680 mg, 5.06 mmol), DIPEA (979 mg, 7.59 mmol), Pd2(dba)3 (231 mg, 0.253 mmol) and XantPhos (219 mg, 0.397 mmol). The reaction mixture was heated to 100°C under nitrogen for 2 hours. The temperature was cooled to room temperature, the solvent was removed under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 2.6 / 1) to obtain the target product 3b (560 mg, solid) with a yield of 89%.

[0172] MS m / z (ESI): 251 [M+1]

[0173] Step 2

[0174] Sodium 1H-indazole-4-thiolate (3c)

[0175] To a solution of 3b (560 mg, 2.24 mmol) in methanol (20 mL) was added a methanol solution of sodium methoxide (30%, 443.5 mg, 2.46 mmol) and stirred at 30°C for 1 hour. The solvent was removed under reduced pressure to obtain the target product 3c (200 mg, solid) with a yield of 51%. The product was used directly in the next step without purification.

[0176] MS m / z (ESI): 151 [M+1]

[0177] Step 3

[0178] ( R )-N-(( R )-8-(5-((1H-indazol-4-yl)thio)-1-methyl-6-carbonyl-1,6-dihydropyrimidin-2-yl)-8-azaspiro[4.5]decane-1-yl)-2-methylpropane-2-sulfenamide (3d)

[0179] To a solution of 3c (70 mg, 0.41 mmol) in dioxane (6 mL) were added 1e (50 mg, 0.10 mmol), DIPEA (37.2 mg, 0.30 mmol), Pd2(dba)3 (9.1 mg, 0.01 mmol) and XantPhos (8.7 mg, 0.015 mmol). The reaction mixture was heated to 80°C under nitrogen protection for 4 hours. The mixture was cooled to room temperature and desolventized under reduced pressure. The residue was purified by silica gel column chromatography (methanol / dichloromethane = 1 / 9) to obtain the target product 3d (50 mg, solid) with a yield of 96%.

[0180] MS m / z (ESI): 515 [M+1]

[0181] Step 4

[0182] ( R )-5-((1H-indazol-4-yl)thio)-2-(1-amino-8-azaspiro[4.5]decane-8-yl)-3-methylpyrimidine-4(3 H )-Ketohydrochloride (3)

[0183] 3d (50 mg, 0.097 mmol) was dissolved in methanol (5 mL), and a solution of hydrogen chloride in dioxane (4.0 M, 5 mL) was added, and the mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure, and the residue was purified by reverse phase preparative high performance liquid chromatography to obtain the target product 3 (16 mg, solid) with a yield of 41%.

[0184] MS m / z (ESI): 411 [M+1]

[0185] 1 H NMR (400 MHz, DMSO- d 6 ) δ 8.14 (brs, 3H), 8.06 (s, 1H), 7.97 (s,1H), 7.40 (d, J= 8.4 Hz, 1H), 7.24 (dd, J = 8.3, 7.2 Hz, 1H), 6.83 (d, J =6.8 Hz, 1H), 3.60 (d, J = 13.5 Hz, 1H), 3.52 (d, J = 13.3 Hz, 1H), 3.40 (s,3H), 3.17 – 3.12 (m, 1H), 3.05 – 2.98 (m, 2H), 2.07 – 2.04 (m, 1H), 1.77 –1.62 (m, 7H), 1.49 (d, J = 12.8 Hz, 1H), 1.39 (d, J = 13.1 Hz, 1H).

[0186] Example 4

[0187] ( R )-2-(1-amino-8-azaspiro[4.5]decane-8-yl)-3-methyl-5-(pyrazolo[1,5- a ]pyridin-4-ylthio)pyrimidine-4(3 H )-Ketoformate

[0188]

[0189] first step

[0190] 4-Bromopyrazolo[1,5- a ]Pyridine-3-carboxylic acid ethyl ester (4b)

[0191] To a solution of 3-bromopyridine 4a (5.0 g, 32.0 mmol) in acetonitrile (25 mL) was added O-(2,4-dinitrophenyl)hydroxylamine (6.37 g, 32.0 mmol), heated to 40°C, and stirred for 16 hours. The solvent was removed under reduced pressure, and the residue was slurried with ether and dried to obtain a yellow solid (3.8 g).

[0192] The above product (2.0 g, 5.6 mmol) was dissolved in DMF (20 mL), potassium carbonate (12.1 g, 88.1 mmol) and ethyl propiolate (7.1 g, 52.8 mmol) were added, and the reaction mixture was stirred at room temperature for 20 hours. Poured into water (50 mL), extracted with ethyl acetate (30 mL×3). The organic phases were combined and washed with saturated brine (20 mL×3), dried over anhydrous sodium sulfate, and desolventized under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 3 / 7) to obtain the target product 4b (350 mg, solid), with a yield of 8%.

[0193] MS m / z (ESI): 269 [M+1]

[0194] Step 2

[0195] 4-Bromopyrazolo[1,5- a ]Pyridine (4c)

[0196] 4b (350 mg, 1.30 mmol) was dissolved in hydrobromic acid solution (12 mL), heated to 100°C, and stirred for 6 hours. The mixture was cooled to room temperature, neutralized with sodium hydroxide solution (2 N), and extracted with ethyl acetate (20 mL×3). The organic phases were combined, washed with saturated brine (15 mL×3), dried over anhydrous sodium sulfate, and desolventized under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 1 / 4) to obtain the target product 4c (200 mg, solid) with a yield of 79%.

[0197] MS m / z (ESI): 197 [M+1]

[0198] Step 3

[0199] 3-(Pyrazolo[1,5- a ]4-pyridin-4-ylthio)propionic acid ethyl ester (4d)

[0200] To a solution of 4c (200 mg, 1.02 mmol) in dioxane (20 mL) were added ethyl 3-mercaptopropionate (680 mg, 5.06 mmol), DIPEA (394 mg, 3.06 mmol), Pd2(dba)3 (93.3 mg, 0.102 mmol) and XantPhos (88.4 mg, 0.153 mmol). The reaction mixture was heated to 100°C under nitrogen for 2 hours. The temperature was cooled to room temperature, the solvent was removed under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 3 / 7) to obtain the target product 4d (240 mg, solid) with a yield of 94%.

[0201] MS m / z (ESI): 251 [M+1]

[0202] Step 4

[0203] Pyrazolo[1,5- a ]Sodium pyridine-4-thiolate (4e)

[0204] To a solution of 4d (230 mg, 0.92 mmol) in methanol (20 mL) was added a methanol solution of sodium methoxide (30%, 182 mg, 1.01 mmol) and stirred at 30°C for 4 hours. The solvent was removed under reduced pressure to obtain the target product 4e (150 mg, solid) with a yield of 95%. The product was used directly in the next step without purification.

[0205] MS m / z (ESI): 151 [M+1]

[0206] Step 5

[0207] ( R )-2-methyl-N-(( R )-8-(1-methyl-6-carbonyl-5-(pyrazolo[1,5- a ]pyridin-4-ylthio)-1,6-dihydropyrimidin-2-yl)-8-azaspiro[4.5]decane-1-yl)propane-2-sulfenamide (4f)

[0208] To a solution of 4e (85.3 mg, 0.495 mmol) in dioxane (6 mL) were added 1e (70 mg, 0.142 mmol), DIPEA (54.9 mg, 0.426 mmol), Pd2(dba)3 (12.9 mg, 0.0142 mmol) and XantPhos (12.3 mg, 0.0213 mmol). The reaction mixture was heated to 80°C under nitrogen protection for 4 hours. The mixture was cooled to room temperature and desolventized under reduced pressure. The residue was purified by silica gel column chromatography (methanol / dichloromethane = 1 / 13) to obtain the target product 4f (40 mg, solid) with a yield of 55%.

[0209] MS m / z (ESI): 515 [M+1]

[0210] Step 6

[0211] ( R )-2-(1-amino-8-azaspiro[4.5]decane-8-yl)-3-methyl-5-(pyrazolo[1,5- a ]pyridin-4-ylthio)pyrimidine-4(3 H )-Ketoformate (4)

[0212] 4f (40 mg, 0.078 mmol) was dissolved in methanol (5 mL), and a solution of hydrogen chloride in dioxane (4.0 M, 5 mL) was added, and the mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure, and the residue was purified by reverse phase preparative HPLC to obtain the target product 4 (16 mg, solid) with a yield of 51%.

[0213] MS m / z (ESI): 411 [M+1]

[0214] 1 H NMR (400 MHz, DMSO- d 6 ) δ 8.56 (d, J = 6.9 Hz, 1H), 8.39 (s, 1H), 8.09 (s, 1H), 8.03 (d, J = 2.2 Hz, 1H), 6.89 (d, J = 7.0 Hz, 1H), 6.80 (t, J = 7.0 Hz, 1H), 6.63 (d, J = 1.7 Hz, 1H), 3.64 – 3.53 (m, 2H), 3.39 (s, 3H), 3.05 – 2.97 (m, 3H), 1.98 – 1.95 (m, 1H), 1.80 – 1.67 (m, 4H), 1.64 – 1.53(m, 3H), 1.40 (d, J = 13.0 Hz, 1H), 1.32 (d, J = 13.3 Hz, 1H).

[0215] Example 5

[0216] ( R )-2-(1-amino-8-azaspiro[4.5]decane-8-yl)-3-methyl-5-(pyrazolo[1,5- a ]pyridin-5-ylthio)pyrimidine-4(3 H )-Ketohydrochloride

[0217]

[0218] first step

[0219] 3-(Pyrazolo[1,5- a ] ethyl pyridin-5-ylthio)propanoate (5b)

[0220] To 5-bromopyrazolo[1,5- a] Ethyl 3-mercaptopropionate (326 mg, 2.43 mmol), DIPEA (627 mg, 4.86 mmol), Pd2(dba)3 (148 mg, 0.162 mmol) and XantPhos (140 mg, 0.243 mmol) were added to a solution of pyridine 5a (320 mg, 1.62 mmol) in dioxane (20 mL). The reaction mixture was heated to 100°C under nitrogen and reacted for 6 hours. The temperature was cooled to room temperature and the solvent was removed under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 3 / 1) to obtain the target product 5b (400 mg, oily substance) with a yield of 97%.

[0221] MS m / z (ESI): 251 [M+1]

[0222] Step 2

[0223] Pyrazolo[1,5- a ]Pyridine-5-thiol (5c)

[0224] To a solution of 5b (400 mg, 1.6 mmol) in methanol (20 mL) was added a methanol solution of sodium methoxide (30%, 317 mg, 1.76 mmol) and stirred at room temperature for 2 hours. The reaction mixture was poured into water (100 mL), the pH was adjusted to weak acidity with citric acid, and extracted with ethyl acetate (30 mL×3). The organic phases were combined and washed with saturated brine (20 mL×3), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and desolventized under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 2 / 3) to obtain the target product 5c (100 mg, oily substance) with a yield of 42%.

[0225] MS m / z (ESI): 151 [M+1]

[0226] Step 3

[0227] ( R )-2-methyl-N-(( R )-8-(1-methyl-6-carbonyl-5-(pyrazolo[1,5- a ]pyridin-5-ylthio)-1,6-dihydropyrimidin-2-yl)-8-azaspiro[4.5]decane-1-yl)propane-2-sulfenamide (5d)

[0228] 5c (30 mg, 0.20 mmol) was dissolved in dioxane (6 mL), and 1e (50 mg, 0.10 mmol), DIPEA (38.7 mg, 0.30 mmol), Pd2(dba)3 (9.0 mg, 0.010 mmol) and XantPhos (8.5 mg, 0.015 mmol) were added. The reaction mixture was heated to 80°C under nitrogen protection and reacted for 2 hours. The mixture was cooled to room temperature and desolventized under reduced pressure. The residue was purified by silica gel column chromatography (methanol / dichloromethane = 1 / 9) to obtain the target product 5d (60 mg, solid, crude product).

[0229] MS m / z (ESI): 515 [M+1]

[0230] Step 4

[0231] ( R )-2-(1-amino-8-azaspiro[4.5]decane-8-yl)-3-methyl-5-(pyrazolo[1,5- a ]pyridin-5-ylthio)pyrimidine-4(3 H )-Ketohydrochloride (5)

[0232] 5d (60 mg, crude product) was dissolved in methanol (5 mL), and a solution of hydrogen chloride in dioxane (4.0 M, 5 mL) was added, and the mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure, and the residue was purified by reverse phase preparative HPLC to obtain the target product 5 (14.1 mg, solid), with a two-step yield of 34%.

[0233] MS m / z (ESI): 411 [M+1]

[0234] 1 H NMR (400 MHz, DMSO- d 6 ) δ 8.56 (d, J = 7.3 Hz, 1H), 8.13 (s, 1H), 8.06 (brs, 3H), 7.95 (s, 1H), 7.94 (d, J = 2.2 Hz, 2H), 7.33 (d, J = 1.4 Hz,1H), 6.68 (dd, J = 7.3, 2.1 Hz, 1H), 6.44 (dd, J = 2.2, 0.7 Hz, 1H), 3.66 (d, J = 13.4 Hz, 1H), 3.58 (d, J= 13.3 Hz, 1H), 3.41 (s, 3H), 3.21 – 3.14 (m,1H), 3.10 – 3.02 (m, 2H), 2.07 – 2.03 (m, 1H), 1.87 – 1.58 (m, 7H), 1.50 (d, J = 12.8 Hz, 1H), 1.41 (d, J = 12.9 Hz, 1H).

[0235] Example 6

[0236] ( R )-5-((1H-indazol-5-yl)thio)-2-(1-amino-8-azaspiro[4.5]decane-8-yl)-3-methylpyrimidine-4(3 H )-Ketohydrochloride

[0237]

[0238] first step

[0239] Ethyl 3-((1H-indazol-5-yl)thio)propanoate (6b)

[0240] Pd2(dba)3 (243 mg, 0.265 mmol) was added to a mixture of 5-bromo-1H-indazole 6a (520 mg, 2.65 mmol), ethyl 3-mercaptopropionate (533 mg, 3.98 mmol), DIPEA (1.02 g, 7.95 mmol), XantPhos (153 mg, 0.265 mmol) and dioxane (10 mL), and the mixture was heated to 100°C under nitrogen protection for 2 hours. The mixture was cooled to room temperature and desolventized under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 1 / 1) to obtain the target product 6b (630 mg, solid) with a yield of 95%.

[0241] MS m / z (ESI): 251 [M+1]

[0242] Step 2

[0243] 1H-Indazole-5-thiol (6c)

[0244] Potassium tert-butoxide (672 mg, 6.0 mmol) was added to a solution of 6b (500 mg, 2.0 mmol) in tetrahydrofuran (10 mL) and stirred at room temperature for 2 hours. The mixture was acidified to pH 4 with hydrochloric acid (6 N) and extracted with dichloromethane (20 mL). The organic phase was dried over anhydrous sodium sulfate, filtered to remove the desiccant, and desolventized under reduced pressure to obtain the target product 6c (300 mg, solid) with a yield of 100%.

[0245] MS m / z (ESI): 151 [M+1]

[0246] Step 3

[0247] ( R )-N-(( R )-8-(5-((1H-indazol-5-yl)thio)-1-methyl-6-carbonyl-1,6-dihydropyrimidin-2-yl)-8-azaspiro[4.5]decane-1-yl)-2-methylpropane-2-sulfenamide (6d)

[0248] Pd2(dba)3 (18 mg, 0.02 mmol) was added to a mixture of 6c (60 mg, 0.4 mmol), 1e (100 mg, 0.2 mmol), DIPEA (77 mg, 0.6 mmol), XantPhos (12 mg, 0.02 mmol) and dioxane (5 mL), and the mixture was heated to 100°C under nitrogen protection for 2 hours. The mixture was cooled to room temperature and desolventized under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 1 / 1) to obtain the target product 6d (70 mg, solid) with a yield of 67%.

[0249] MS m / z (ESI): 515 [M+1]

[0250] Step 4

[0251] ( R )-5-((1H-indazol-5-yl)thio)-2-(1-amino-8-azaspiro[4.5]decane-8-yl)-3-methylpyrimidine-4(3 H )-Ketohydrochloride (6)

[0252] 6d (70 mg, 0.14 mmol) was dissolved in methanol (5 mL), and a solution of hydrogen chloride in dioxane (4.0 M, 2 mL) was added, and the mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure, and the residue was purified by reverse phase preparative HPLC to obtain the target product 6 (30 mg, solid) with a yield of 54%.

[0253] MS m / z (ESI): 411 [M+1]

[0254] 1 H NMR (400 MHz, DMSO- d 6 ) δ 8.28 (s, 3H), 8.05 (s, 1H), 7.79 (s, 1H), 7.69 (s, 1H), 7.55 (d, J = 8.7 Hz, 1H), 7.34 (dd, J = 8.7, 1.6 Hz, 1H), 3.56(d, J = 13.2 Hz, 1H), 3.49 (d, J = 13.5 Hz, 1H), 3.39 (s, 3H), 3.17 – 3.13(m, 1H), 3.06 – 2.96 (m, 2H), 2.07 – 2.01 (m, 1H), 1.90 – 1.61 (m, 7H), 1.52(d, J = 13.1 Hz, 1H), 1.39 (d, J = 13.0 Hz, 1H).

[0255] Example 7

[0256] ( R )-2-(1-amino-8-azaspiro[4.5]decane-8-yl)-5-((4-chloro-2-methyl-2H-indazol-5-yl)thio)-3-methylpyrimidine-4(3 H )-Ketohydrochloride

[0257]

[0258] first step

[0259] 5-Bromo-4-chloro-2H-indazole (7b)

[0260] Under ice-bath cooling, sodium nitrite (396 mg, 5.73 mmol) in water (4 mL) was added to a solution of 4-bromo-3-chloro-2-methylaniline 7a (1.0 g, 4.58 mmol) in acetic acid (20 mL), and the mixture was stirred at room temperature for 1 hour. Most of the solvent was removed by rotary evaporation, and the residue was suspended in water (30 mL) and filtered. The filter cake was washed with water (25 mL×3) and dried in air to obtain the target product 7b (780 mg, solid) with a yield of 74%. The product was used directly in the next step without purification.

[0261] MS m / z (ESI): 231 [M+1]

[0262] Step 2

[0263] 5-Bromo-4-chloro-2-methyl-2H-indazole (7c)

[0264] Trimethyloxonium tetrafluoroborate (733 mg, 4.95 mmol) was added to a solution of 7b (760 mg, 3.3 mmol) in ethyl acetate (15 mL) under ice-bath cooling, and the reaction mixture was stirred at room temperature for 4 hours. Petroleum ether (30 mL) was added to dilute, stirred for 10 minutes, and filtered. Ethyl acetate (20 mL) was added to the filtrate, and washed with sodium bicarbonate solution (30 mL×3) and saturated brine (20 mL×3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 1.4 / 1) to obtain the target product 7c (460 mg, solid) with a yield of 57%.

[0265] MS m / z (ESI): 245 [M+1]

[0266] Step 3

[0267] Ethyl 3-((4-chloro-2-methyl-2H-indazol-5-yl)thio)propanoate (7d)

[0268] To a solution of 7c (440 mg, 1.8 mmol) in dioxane (20 mL) were added ethyl 3-mercaptopropionate (483 mg, 3.6 mmol), DIPEA (697 mg, 5.4 mmol), Pd2(dba)3 (165 mg, 0.18 mmol) and XantPhos (156 mg, 0.27 mmol). The reaction mixture was heated to 100°C under nitrogen and reacted for 16 hours. The temperature was cooled to room temperature and the solvent was removed under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 1.7 / 1) to obtain the target product 7d (300 mg, solid) with a yield of 59%.

[0269] MS m / z (ESI): 299 [M+1]

[0270] Step 4

[0271] Sodium 4-chloro-2-methyl-2H-indazole-5-thiolate (7e)

[0272] To a solution of 7d (140 mg, 0.47 mmol) in methanol (20 mL) was added a methanol solution of sodium methoxide (30%, 93 mg, 0.52 mmol) and stirred at 30°C for 32 hours. The solvent was removed under reduced pressure to obtain the target product 7e (100 mg, solid) with a yield of 96%. The product was used directly in the next step without purification.

[0273] MS m / z (ESI): 199 [M+1]

[0274] Step 5

[0275] ( R )-N-(( R )-8-(5-((4-chloro-2-methyl-2H-indazol-5-yl)thio)-1-methyl-6-carbonyl-1,6-dihydropyrimidin-2-yl)-8-azaspiro[4.5]decane-1-yl)-2-methylpropane-2-sulfinamide (7f)

[0276] 7e (80 mg, 0.36 mmol) was dissolved in dioxane (8 mL), and 1e (100 mg, 0.20 mmol), DIPEA (77 mg, 0.60 mmol), Pd2(dba)3 (18 mg, 0.020 mmol) and XantPhos (18 mg, 0.030 mmol) were added. The reaction mixture was heated to 90°C under nitrogen protection and reacted for 2 hours. After cooling to room temperature, the reaction mixture was poured into a mixed solvent of ethyl acetate (15 mL) and petroleum ether (15 mL), filtered, and the filter cake was dried to obtain the target product 7f (61 mg, solid) with a yield of 54%.

[0277] MS m / z (ESI): 563 [M+1]

[0278] Step 6

[0279] ( R )-2-(1-amino-8-azaspiro[4.5]decane-8-yl)-5-((4-chloro-2-methyl-2H-indazol-5-yl)thio)-3-methylpyrimidine-4(3 H )-Ketohydrochloride (7)

[0280] 7f (61 mg, 0.11 mmol) was dissolved in methanol (5 mL), and a solution of hydrogen chloride in dioxane (4.0 M, 5 mL) was added, and the mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure, and the residue was purified by reverse phase preparative HPLC to obtain the target product 7 (16 mg, solid) with a yield of 31%.

[0281] MS m / z (ESI): 459 [M+1]

[0282] 1 H NMR (400 MHz, DMSO- d 6) δ 8.43 (s, 1H), 8.12 (brs, 3H), 7.90 (s,1H), 7.51 (dd, J = 9.0, 0.7 Hz, 1H), 6.99 (d, J = 9.0 Hz, 1H), 4.17 (s, 3H), 3.60 (d, J = 13.3 Hz, 1H), 3.53 (d, J = 13.3 Hz, 1H), 3.40 (s, 3H), 3.16 –3.13 (m, 1H), 3.06 – 2.98 (m, 2H), 2.07 – 2.04 (m, 1H), 1.87 – 1.60 (m, 7H),1.49 (d, J = 13.0 Hz, 1H), 1.39 (d, J = 13.1 Hz, 1H).

[0283] Example 8

[0284] ( R )-2-(1-amino-8-azaspiro[4.5]decane-8-yl)-5-((4-chloropyrazolo[1,5- a ]pyridin-5-yl)thio)-3-methylpyrimidine-4(3 H )-Ketoformate

[0285]

[0286] first step

[0287] Tert-butyl [[(2,4,6-trimethylphenyl)sulfonyl]oxy]carbamate (8b)

[0288] 2,4,6-Trimethylbenzenesulfonyl chloride 8a (2.0 g, 9.15 mmol) and tert-butyl hydroxycarbamate (1.22 g, 9.15 mmol) were dissolved in tert-butyl methyl ether (30 mL), cooled to 0°C, triethylamine (1.4 mL, 10.1 mmol) was added dropwise, and then warmed to room temperature and stirred for 4 hours. After washing with water, the organic phase was dried over anhydrous sodium sulfate and desolventized under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 1 / 5) to obtain the target product 8b (2.5 g, solid) with a yield of 87%.

[0289] MS m / z (ESI): 338 [M+23], 216 [M+1-100]

[0290] Step 2

[0291] O-(2,4,6-Trimethylbenzenesulfonyl)hydroxylamine (8c)

[0292] 8b (2.5 g, 7.93 mmol) was added in portions to trifluoroacetic acid (20 mL) at 0°C, and the reaction mixture was stirred at 0°C for 2 hours. Ice water was slowly added and stirred for 15 minutes. Filtered, the filter cake was washed with water until the filtrate was neutral. The filter cake was dried to obtain the target product 8c (1.5 g, solid), with a yield of 88%.

[0293] MS m / z (ESI): 216 [M+1]

[0294] Step 3

[0295] 5-Bromo-4-chloropyrazolo[1,5- a ]Pyridine-3-carboxylic acid ethyl ester (8e)

[0296] 4-Bromo-3-chloropyridine 8d (1.4 g, 7.28 mmol) was dissolved in acetonitrile (25 mL), 8c (1.6 g, 7.28 mmol) was added, and the reaction mixture was stirred at 40°C overnight. The product was filtered and dried to obtain a yellow solid product (1.7 g).

[0297] The above product (1.6 g, 3.92 mmol) and ethyl propiolate (0.42 g, 4.32 mmol) were dissolved in N,N -dimethylformamide (15 mL), potassium carbonate (1.1 g, 7.84 mmol) was added, and stirred at room temperature for 3 hours. Water was added to quench the mixture, and the mixture was extracted with ethyl acetate (100 mL×3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and desolventized under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 1 / 2) to obtain the target product 8e (120 mg, solid), with a yield of 5.8%.

[0298] MS m / z (ESI): 303, 305 [M+1]

[0299] Step 4

[0300] 5-Bromo-4-chloropyrazolo[1,5- a ]Pyridine (8f)

[0301] 8e (110 mg, 0.36 mmol) was dissolved in hydrobromic acid solution (3 mL), heated to 100°C, and stirred for 3 hours. The mixture was cooled to room temperature, neutralized to neutrality by adding sodium hydroxide solution (2 M), and extracted with dichloromethane (30 mL×3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and desolventized under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 3 / 7) to obtain the target product 8f (65 mg, solid), with a yield of 77%.

[0302] MS m / z (ESI): 231, 233 [M+1]

[0303] Step 5

[0304] 3-((4-chloropyrazolo[1,5- a ]Ethyl pyridin-5-yl)thio)propanoate (8 g)

[0305] To a solution of 8f (60 mg, 0.26 mmol) in dioxane (5 mL) were added ethyl 3-mercaptopropionate (38 mg, 0.29 mmol), DIPEA (67 mg, 0.52 mmol), Pd2(dba)3 (24 mg, 0.026 mmol) and XantPhos (30 mg, 0.052 mmol). The reaction mixture was heated to 100°C under nitrogen for 3 hours. The mixture was cooled to room temperature and desolventized under reduced pressure. The residue was purified by silica gel column chromatography (methanol / dichloromethane = 1 / 28) to obtain the target product 8g (60 mg, solid) with a yield of 81%.

[0306] MS m / z (ESI): 285 [M+1]

[0307] Step 6

[0308] 4-Chloropyrazolo[1,5- a ]Sodium pyridine-5-thiolate (8h)

[0309] To a solution of 8 g (60 mg, 0.21 mmol) in methanol (3 mL), add a methanol solution of sodium methoxide (30%, 45.5 mg, 0.25 mmol) and stir at 30°C for 4 hours. Remove the solvent under reduced pressure to obtain the target product 8h (40 mg, solid) with a yield of 92%. The product was used directly in the next reaction without purification.

[0310] MS m / z (ESI): 185 [M+1]

[0311] Step 7

[0312] ( R )-N-((R )-8-(5-((4-chloropyrazolo[1,5- a ]pyridin-5-yl)thio)-1-methyl-6-carbonyl-1,6-dihydropyrimidin-2-yl)-8-azaspiro[4.5]decane-1-yl)-2-methylpropane-2-sulfinamide (8i)

[0313] 8h (40 mg, 0.19 mmol) was dissolved in dioxane (5 mL), and 1e (95 mg, 0.19 mmol), DIPEA (49 mg, 0.38 mmol), Pd2(dba)3 (17 mg, 0.02 mmol) and XantPhos (22 mg, 0.04 mmol) were added. The reaction mixture was heated to 100°C under nitrogen protection and reacted for 4 hours. The mixture was cooled to room temperature and desolventized under reduced pressure. The residue was purified by silica gel column chromatography (methanol / dichloromethane = 1 / 28) to obtain the target product 8i (32 mg, oily substance) with a yield of 30%.

[0314] MS m / z (ESI): 549 [M+1]

[0315] Step 8

[0316] ( R )-2-(1-amino-8-azaspiro[4.5]decane-8-yl)-5-((4-chloropyrazolo[1,5- a ]pyridin-5-yl)thio)-3-methylpyrimidine-4(3 H )-Ketoformate (8)

[0317] 8i (32 mg, 0.06 mmol) was dissolved in methanol (2 mL), and a solution of hydrogen chloride in dioxane (4.0 M, 1 mL) was added, and the mixture was stirred at room temperature for 1 hour. The mixture was concentrated under reduced pressure, and the residue was purified by reverse phase preparative HPLC to obtain the target product 8 (13 mg, solid) with a yield of 47%.

[0318] MS m / z (ESI): 445 [M+1]

[0319] 1 H NMR (400 MHz, DMSO- d 6 ) δ 8.54 (d, J = 7.3 Hz, 1H), 8.35 (s, 1H), 8.19 (s, 1H), 8.06 (d, J = 2.2 Hz, 1H), 6.60 (d, J = 1.8 Hz, 1H), 6.48 (d,J = 7.3 Hz, 1H), 3.69 – 3.60 (m, 2H), 3.41 (s, 3H), 3.09 (t, J = 11.7 Hz, 2H), 3.02 – 2.99 (m, 1H), 2.00 – 1.91 (m, 1H), 1.76 – 1.50 (m, 7H), 1.42 (d, J =13.2 Hz, 1H), 1.35 (d, J = 12.9 Hz, 1H).

[0320] Example 9

[0321] ( R )-5-((2-(1-amino-8-azaspiro[4.5]decane-8-yl)-1-methyl-6-carbonyl-1,6-dihydropyrimidin-5-yl)thio)-4-chloro-2-methyl-2H-indazole-3-carbonitrile

[0322]

[0323] first step

[0324] 5-Bromo-4-chloro-2-methyl-2H-indazole-3-carbaldehyde (9a)

[0325] 7c (800 mg, 3.28 mmol) was dissolved in tetrahydrofuran (20 mL), cooled to -78°C, and a solution of lithium diisopropylamide in tetrahydrofuran (2 M, 2.95 mL, 5.9 mmol) was added. The mixture was stirred for 90 minutes, and the mixture was heated to 0°C and stirred for 30 minutes. The mixture was cooled to -78°C again, and DMF (0.76 mL) was added dropwise. The mixture was stirred for 0.5 hours, and the mixture was heated to room temperature and stirred for 2 hours. The mixture was quenched by adding saturated ammonium chloride solution and extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, filtered to remove the desiccant, and desolventized under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 3 / 7) to obtain the target product 9a (520 mg, solid) with a yield of 60%.

[0326] MS m / z (ESI): 273 [M+1]

[0327] Step 2

[0328] 5-Bromo-4-chloro-2-methyl-2H-indazole-3-carbaldehyde oxime (9b)

[0329] 9a (490 mg, 1.8 mmol) was dissolved in isopropanol (8 mL), methanol (8 mL) and water (8 mL), and hydroxylamine hydrochloride (497 mg, 7.2 mmol) and sodium carbonate (763 mg, 7.2 mmol) were added, heated to 50°C, and stirred for 16 hours. Poured into water (60 mL), extracted with ethyl acetate (50 mL×3). The organic phases were combined and washed with saturated brine (20 mL×3), dried over anhydrous sodium sulfate, and desolventized under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 3 / 7) to obtain the target product 9b (470 mg, solid), with a yield of 92%.

[0330] MS m / z (ESI): 288 [M+1]

[0331] Step 3

[0332] 5-Bromo-4-chloro-2-methyl-2H-indazole-3-carbonitrile (9c)

[0333] 9b (470 mg, 1.63 mmol) was dissolved in acetonitrile (30 mL), copper acetate (147 mg, 0.81 mmol) was added, heated to 85°C, and stirred overnight. The solvent was removed under reduced pressure, and the residue was poured into water (50 mL) and extracted with ethyl acetate (30 mL × 3). The organic phases were combined and washed with saturated brine (25 mL × 3), dried over anhydrous sodium sulfate, and removed under reduced pressure to obtain the target product 9c (390 mg, solid) with a yield of 89%.

[0334] MS m / z (ESI): 270 [M+1]

[0335] Step 4

[0336] Ethyl 3-((4-chloro-3-cyano-2-methyl-2H-indazol-5-yl)thio)propanoate (9d)

[0337] To a solution of 9c (380 mg, 1.4 mmol) in dioxane (5 mL) were added ethyl 3-mercaptopropionate (385 mg, 2.8 mmol), DIPEA (542 mg, 4.2 mmol), Pd2(dba)3 (128 mg, 0.14 mmol) and XantPhos (121 mg, 0.21 mmol). The reaction mixture was heated to 100°C under nitrogen protection and reacted for 16 hours. The temperature was cooled to room temperature and the solvent was removed under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 1 / 1) to obtain the target product 9d (350 mg, solid) with a yield of 77%.

[0338] MS m / z (ESI): 324 [M+1]

[0339] Step 5

[0340] Sodium 4-chloro-3-cyano-2-methyl-2H-indazole-5-thiolate (9e)

[0341] To a solution of 9d (195 mg, 0.60 mmol) in methanol (20 mL) was added a methanol solution of sodium methoxide (30%, 130 mg, 0.72 mmol), and the mixture was stirred at 30°C for 16 hours. The solvent was removed under reduced pressure to obtain the target product 9e (220 mg, solid, crude product). The product was used directly in the next step without purification.

[0342] MS m / z (ESI): 224 [M+1]

[0343] Step 6

[0344] ( R )-N-(( R )-8-(5-((4-chloro-3-cyano-2-methyl-2H-indazol-5-yl)thio)-1-methyl-6-carbonyl-1,6-dihydropyrimidin-2-yl)-8-azaspiro[4.5]decane-1-yl)-2-methylpropane-2-sulfinamide (9f)

[0345] 9e (89.2 mg, 0.36 mmol) was dissolved in dioxane (6 mL), and 1e (120 mg, 0.24 mmol), DIPEA (92.8 mg, 0.72 mmol), Pd2(dba)3 (21.9 mg, 0.024 mmol) and XantPhos (20.8 mg, 0.036 mmol) were added. The reaction mixture was heated to 80°C under nitrogen protection and reacted for 16 hours. The mixture was cooled to room temperature and desolventized under reduced pressure. The residue was purified by silica gel column chromatography (methanol / dichloromethane = 1 / 12) to obtain the target product 9f (90 mg, solid) with a two-step yield of 63%.

[0346] MS m / z (ESI): 588 [M+1]

[0347] Step 7

[0348] ( R )-5-((2-(1-amino-8-azaspiro[4.5]decane-8-yl)-1-methyl-6-carbonyl-1,6-dihydropyrimidin-5-yl)thio)-4-chloro-2-methyl-2H-indazole-3-carbonitrile formate (9)

[0349] 9f (90 mg, 0.15 mmol) was dissolved in methanol (10 mL), and a solution of hydrogen chloride in dioxane (4.0 M, 10 mL) was added, and the mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure, and the residue was purified by reverse phase preparative HPLC to obtain the target product 9 (14 mg, solid) with a yield of 19%.

[0350] MS m / z (ESI): 484 [M+1]

[0351] 1 H NMR (400 MHz, DMSO- d 6 ) δ 8.14 (s, 1H), 7.79 (brs, 3H), 7.75 (d, J =9.0 Hz, 1H), 7.05 (d, J = 9.1 Hz, 1H), 4.34 (s, 3H), 3.69 (d, J = 14.1 Hz,1H), 3.61 (d, J = 14 Hz, 1H), 3.41 (s, 3H), 3.24 – 3.16 (m, 1H), 3.13 – 3.06(m, 2H), 2.08 – 2.04 (m, 1H), 1.80 – 1.64 (m, 7H), 1.49 – 1.41 (m, 2H).

[0352] Example 10

[0353] ( S )-2-(4-amino-2-oxa-8-azaspiro[4.5]decane-8-yl)-5-((4-chloro-2-methyl-2H-indazol-5-yl)thio)-3-methylpyrimidine-4(3 H )-ketone

[0354]

[0355] first step

[0356] 2-((tert-Butyldimethylsilyl)oxy)ethan-1-ol (10b)

[0357] Tert-butyldimethylsilyl chloride (50 g, 334 mmol) was added dropwise to a mixture of ethylene glycol 10a (124 g, 2 mol), imidazole (34 g, 500 mmol) and dichloromethane (500 mL) at 0°C, and the reaction mixture was stirred at room temperature for 16 hours. Water (300 mL) and dichloromethane (600 mL) were added to dilute the mixture, and the liquids were separated. The aqueous phase was extracted with dichloromethane (300 mL×3). The organic phases were combined and dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 1 / 5) to obtain the target product 10b (35 g, oily substance) with a yield of 10%.

[0358] MS m / z (ESI): 177 [M+1]

[0359] Step 2

[0360] 2-((tert-Butyldimethylsilyl)oxy)acetaldehyde (10c)

[0361] Oxalyl chloride (27.6 g, 218.9 mmol) was added dropwise to dichloromethane (500 mL) at -30°C, cooled to -78°C, and dimethyl sulfoxide (21.7 g, 278.6 mmol) was added dropwise. After the reaction mixture was stirred at -78°C for 30 minutes, a solution of 10b (35 g, 199 mmol) in dichloromethane (100 mL) was slowly added and stirred for 1 hour. Triethylamine (100.5 g, 995 mmol) was added dropwise, and stirring was continued at -78°C for 30 minutes, and then stirred at room temperature overnight. The mixture was washed with water (300 mL), dilute hydrochloric acid (1N, 400 mL×2), saturated sodium bicarbonate solution (400 mL), and saturated brine (400 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the target product 10c (34 g, oily substance) with a yield of 98%.

[0362] 1 H NMR (400 MHz, CDCl3) δ 9.60 (s, 1H), 4.11 (s, 2H), 0.82 (s, 9H), 0.00 (s, 6H).

[0363] Step 3

[0364] 1-(tert-Butyl)4-ethyl 4-(2-((tert-butyldimethylsilyl)oxy)-1-hydroxyethyl)piperidine-1,4-dicarboxylate (10d)

[0365] To a tetrahydrofuran solution of lithium diisopropylamide (2 M, 148 mL, 296 mmol) and tetrahydrofuran (400 mL) was added dropwise a solution of 1-(tert-butyl) 4-ethylpiperidine-1,4-dicarboxylate (50 g, 197 mmol) in tetrahydrofuran (100 mL) at -10°C, and the reaction mixture was stirred at 0°C for 30 minutes. 10c (34 g, 197 mmol) was added, and the mixture was stirred at 0°C for 1 hour and at room temperature for 1 hour. A mixed solution of saturated sodium bicarbonate and water (400 mL, 1 / 4 v / v) was added, and ethyl acetate (200 mL) was added, and the layers were separated. The aqueous phase was extracted with ethyl acetate (200 mL×3). The organic phases were combined and dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 2 / 3) to obtain the target product 10d (75 g, oily substance) with a yield of 88%.

[0366] MS m / z (ESI): 332 [M+1-100]

[0367] Step 4

[0368] tert-Butyl 4-(2-((tert-butyldimethylsilyl)oxy)-1-hydroxyethyl)-4-(hydroxymethyl)piperidine-1-carboxylate (10e)

[0369] To a solution of 10d (75 g, 174 mmol) in tetrahydrofuran (600 mL) was added a solution of lithium borohydride in tetrahydrofuran (2 M, 130 mL, 261 mmol) and stirred at room temperature for 2 hours. The mixture was cooled to 0°C, quenched with saturated sodium bicarbonate solution and water (150 mL, 1 / 2 v / v), diluted with ethyl acetate (200 mL), and filtered. The filtrate was separated, and the aqueous phase was extracted with ethyl acetate (200 mL×3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the target product 10e (55 g, oily substance) with a yield of 81%.

[0370] MS m / z (ESI): 290 [M+1-100]

[0371] Step 5

[0372] 4-(1,2-Dihydroxyethyl)-4-(hydroxymethyl)piperidine-1-carboxylic acid tert-butyl ester (10f)

[0373] To a solution of 10e (55 g, 128.5 mmol) in tetrahydrofuran (400 mL) was added a solution of tetrabutylammonium fluoride in tetrahydrofuran (1 M, 193 mL, 193 mmol) and stirred at room temperature for 2 hours. The mixture was diluted with a saturated sodium bicarbonate solution and water (100 mL, 1 / 2 v / v), ethyl acetate (200 mL) was added, the liquids were separated, and the aqueous phase was extracted with ethyl acetate (200 mL×3). The organic phases were combined and dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 1 / 4 to 1 / 0) to obtain the target product 10f (23 g, oily substance) with a yield of 65%.

[0374] MS m / z (ESI): 176 [M+1-100]

[0375] Step 6

[0376] 4-Hydroxy-2-oxa-8-azaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (10 g)

[0377] Sodium hydride (60%, 11.7 g, 292.6 mmol) was suspended in tetrahydrofuran (100 mL), cooled to 0°C, and a solution of 10f (23 g, 83.6 mmol) in tetrahydrofuran (100 mL) was added dropwise, followed by a solution of 4-toluenesulfonyl chloride (16 g, 83.6 mmol) in tetrahydrofuran (200 mL). The reaction mixture was stirred at 0°C for 2 hours. Saturated ammonium chloride solution (50 mL) was slowly added and stirred vigorously until no gas was generated. Saturated ammonium chloride solution (100 mL) and saturated brine (100 mL) were added and extracted with ethyl acetate (200 mL×3). The organic phases were combined and dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 0 / 1 to 1 / 0) to obtain the target product 10g (8.5 g, oily substance) with a yield of 39%.

[0378] MS m / z (ESI): 158 [M+1-100]

[0379] Step 7

[0380] 4-Carbonyl-2-oxa-8-azaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (10h)

[0381] The reaction mixture 10g (8.5 g, 32.9 mmol), Dess-Martin periodinane (27.4 g, 64.56 mmol) and dichloromethane (200 mL) were stirred at 0°C for 2 hours. Saturated sodium bicarbonate solution and saturated sodium thiosulfate solution (100 mL, 1 / 1 v / v) were added, stirred vigorously, and the liquids were separated. The aqueous phase was extracted with dichloromethane (200 mL×3). The organic phases were combined and dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 2 / 3) to obtain the target product 10h (6.2 g, oily substance) with a yield of 74%.

[0382] MS m / z (ESI): 156 [M+1-100]

[0383] Step 8

[0384] 2-Oxa-8-azaspiro[4.5]decan-4-one (10i)

[0385] To a solution of 10h (500 mg, 1.96 mmol) in dichloromethane (8 mL) was added a solution of hydrogen chloride in dioxane (4 M, 2 mL) and stirred at room temperature for 1 hour. The solvent was removed under reduced pressure to obtain the target product 10i (hydrochloride, 370 mg, solid) with a yield of 99%. The product was used directly in the next step without purification.

[0386] MS m / z (ESI): 156 [M+1]

[0387] Step 9

[0388] 8-(5-iodo-1-methyl-6-carbonyl-1,6-dihydropyrimidin-2-yl)-2-oxa-8-azaspiro[4.5]decan-4-one (10j)

[0389] 1c (100 mg, 0.37 mmol) was added to a mixture of 10i (hydrochloride, 106 mg, 0.555 mmol), potassium carbonate (153 mg, 1.11 mmol) and acetonitrile (4 mL), heated to 80 °C and stirred for 16 h. The mixture was cooled to room temperature, and water (5 mL) and ethyl acetate (20 mL) were added and separated. The aqueous phase was extracted with ethyl acetate (10 mL×3). The organic phases were combined and dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 1 / 4) to obtain the target product 10j (70 mg, solid) with a yield of 49%.

[0390] MS m / z (ESI): 390 [M+1]

[0391] Step 10

[0392] ( R )-N-(( S )-8-(5-iodo-1-methyl-6-carbonyl-1,6-dihydropyrimidin-2-yl)-2-oxa-8-azaspiro[4.5]decane-4-yl)-2-methylpropane-2-sulfenamide (10k)

[0393] The reaction mixture 10j (60 mg, 0.15 mmol), ( R )-2-methylpropane-2-sulfenamide (36.3 mg, 0.3 mmol), tetraethyl titanate (137 mg, 0.6 mmol) and tetrahydrofuran (5 mL) were heated to 90°C and stirred for 4 hours. After cooling to 0°C, methanol (2 mL) and a tetrahydrofuran solution of lithium borohydride (2.0 M, 0.15 mL, 0.3 mmol) were added and stirred for 1 hour. Water (5 mL) and dichloromethane (20 mL) were added and the mixture was separated. The aqueous phase was extracted with dichloromethane (10 mL×3). The organic phases were combined and dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (methanol / dichloromethane = 1 / 9) to obtain the target product 10k (70 mg, solid) with a yield of 92%.

[0394] MS m / z (ESI): 495 [M+1]

[0395] Step 11

[0396] ( R )-N-(( S )-8-(5-((4-chloro-2-methyl-2H-indazol-5-yl)thio)-1-methyl-6-carbonyl-1,6-dihydropyrimidin-2-yl)-2-oxa-8-azaspiro[4.5]decane-4-yl)-2-methylpropane-2-sulfenamide (10l)

[0397] Pd2(dba)3 (12.8 mg, 0.014 mmol) was added to a mixture of 10k (70 mg, 0.14 mmol), 7e (37.4 mg, 0.17 mmol), DIPEA (54 mg, 0.42 mmol), XantPhos (8.1 mg, 0.014 mmol) and dioxane (4 mL), and the mixture was heated to 100°C under nitrogen protection for 2 hours. The mixture was cooled to room temperature, and water (5 mL) and dichloromethane (20 mL) were added and separated. The aqueous phase was extracted with dichloromethane (10 mL×3), and the organic phases were combined and dried over anhydrous sodium sulfate, filtered, and desolventized under reduced pressure. The residue was purified by silica gel column chromatography (methanol / dichloromethane = 1 / 9) to obtain the target product 10l (50 mg, solid) with a yield of 63%.

[0398] MS m / z (ESI): 565 [M+1]

[0399] Step 12

[0400] ( S )-2-(4-amino-2-oxa-8-azaspiro[4.5]decane-8-yl)-5-((4-chloro-2-methyl-2H-indazol-5-yl)thio)-3-methylpyrimidine-4(3 H )-Keto (10)

[0401] 10l (50 mg, 0.09 mmol) was dissolved in methanol (8 mL), and a solution of hydrogen chloride in dioxane (4.0 M, 2 mL) was added, and the mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure, and the residue was purified by reverse phase preparative HPLC to obtain the target product 10 (5.8 mg, solid) with a yield of 14%.

[0402] MS m / z (ESI): 461 [M+1]

[0403] 1 H NMR (400 MHz, DMSO- d 6 ) δ 8.43 (s, 1H), 7.91 (s, 1H), 7.51 (d, J =9.0 Hz, 1H), 6.99 (d, J = 9.0 Hz, 1H), 4.18 (s, 3H), 4.02 – 3.95 (m, 1H), 3.69 (d, J = 8.5 Hz, 1H), 3.63 (d, J= 8.5 Hz, 1H), 3.39 (s, 3H), 3.15 – 3.13(m, 1H), 3.08 – 3.00 (m, 2H), 1.84 – 1.76 (m, 1H), 1.72 – 1.65 (m, 1H), 1.54– 1.43 (m, 2H).

[0404] Embodiment 11

[0405] 2-((3 S ,4 S )-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decane-8-yl)-5-((4-chloro-2-methyl-2H-indazol-5-yl)thio)-3-methylpyrimidine-4(3 H )-Ketoformate

[0406]

[0407] first step

[0408] ( S )-2-((tert-butyldimethylsilyl)oxy)propionic acid methyl ester (11b)

[0409] To a solution of imidazole (68.5 g, 1008 mmol) in dichloromethane (800 mL) was added dropwise at 0°C ( S )-2-hydroxypropionic acid methyl ester 11a (70 g, 672 mmol), then slowly add tert-butyldimethylsilyl chloride (120 g, 807 mmol), and the reaction mixture is stirred at room temperature for 16 hours. Dilute with water (600 mL) and extract with dichloromethane (500 mL×2). The organic phases are combined and washed with dilute hydrochloric acid (1 N, 300 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the target product 11b (150 g, oily substance) with a yield of 99%.

[0410] 1 H NMR (400 MHz, CDCl3) δ 4.32 (q, J = 6.7 Hz, 1H), 3.70 (s, 3H), 1.38(d, J = 6.8 Hz, 3H), 0.88 (s, 9H), 0.08 (s, 3H), 0.05 (s, 3H).

[0411] Step 2

[0412] ( S)-2-((tert-Butyldimethylsilyl)oxy)propanal (11c)

[0413] A solution of 11b (151.2 g, 693 mmol) in dichloromethane (1200 mL) was cooled to -78°C, and a solution of diisobutylaluminum hydride in n-hexane (1 M, 832 mL, 832 mmol) was added dropwise. The mixture was heated to -40°C and stirred for 1 hour. The reaction mixture was poured into a saturated solution of sodium potassium tartrate (140 mL), and ether (100 mL) was added. The mixture was stirred at room temperature for 2 hours. The liquids were separated, and the aqueous phase was extracted with ether. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 4 / 1) to obtain the target product 11c (62.3 g, oily substance) with a yield of 48%.

[0414] 1 H NMR (400 MHz, CDCl3) δ 9.60 (s, 1H), 4.09 (q, J = 6.8 Hz, 1H), 1.27(d, J = 6.9 Hz, 3H), 0.91 (s, 9H), 0.10 (s, 3H), 0.08 (s, 3H).

[0415] Step 3

[0416] 1-(tert-butyl)4-ethyl4-((2 S )-2-((tert-Butyldimethylsilyl)oxy)-1-hydroxypropyl)piperidine-1,4-dicarboxylate (11d)

[0417] To a tetrahydrofuran solution of lithium diisopropylamide (1 M, 425 mL, 425 mmol) and tetrahydrofuran (800 mL) was added dropwise a solution of 1-(tert-butyl) 4-ethylpiperidine-1,4-dicarboxylate (72.8 g, 283.5 mmol) in tetrahydrofuran (50 mL) at -50°C, and the reaction mixture was stirred at -10°C for 1 hour. 11c (53.3 g, 283.5 mmol) was added and stirred at -10°C for 1 hour. The temperature was raised to 0°C and stirred for 1 hour, and then stirred at room temperature for 1 hour. A mixed solution of saturated sodium bicarbonate and water (400 mL, 1 / 4 v / v) was added, and ethyl acetate (500 mL) was added and the layers were separated. The aqueous phase was extracted with ethyl acetate (500 mL×3). The organic phases were combined and dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the target product 11d (120 g, oil, crude product). The product was used directly in the next reaction without purification.

[0418] MS m / z (ESI): 346 [M+1-100]

[0419] Step 4

[0420] 4-((2 S )-2-((tert-butyldimethylsilyl)oxy)-1-hydroxypropyl)-4-(hydroxymethyl)piperidine-1-carboxylic acid tert-butyl ester (11e)

[0421] To a solution of 11d (120 g, 269.6 mmol) in tetrahydrofuran (1200 mL) was added a solution of lithium borohydride in tetrahydrofuran (2 M, 270 mL, 539.2 mmol) and stirred at room temperature for 16 hours. The mixture was cooled to 0°C, quenched with saturated sodium bicarbonate solution and water (600 mL, 1 / 2 v / v), diluted with ethyl acetate (800 mL), and filtered. The filtrate was separated, and the aqueous phase was extracted with ethyl acetate (500 mL×3). The organic phases were combined and washed with saturated brine (300 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the target product 11e (90 g, oily, crude). The product was used directly in the next step without purification.

[0422] MS m / z (ESI): 404 [M+1]

[0423] Step 5

[0424] 4-((2 S tert-Butyl 1,2-dihydroxypropyl)-4-(hydroxymethyl)piperidine-1-carboxylate (11f)

[0425] To a solution of 11e (90 g, 223 mmol) in tetrahydrofuran (1300 mL) was added a tetrahydrofuran solution of tetrabutylammonium fluoride (1 M, 303 mL, 303 mmol) and stirred at room temperature overnight. The mixture was diluted with saturated sodium bicarbonate solution and water (800 mL, 1 / 2 v / v), ethyl acetate (800 mL) was added, the liquids were separated, and the aqueous phase was extracted with ethyl acetate (500 mL×3). The organic phases were combined and washed with saturated brine (300 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 49 / 1) to obtain the target product 11f (24.5 g, oily substance), with a three-step yield of 26%.

[0426] MS m / z (ESI): 290 [M+1]

[0427] Step 6

[0428] (3 S)-4-hydroxy-3-methyl-2-oxa-8-azaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (11 g)

[0429] Sodium hydride (60%, 10.8 g, 270.0 mmol) was suspended in tetrahydrofuran (600 mL), cooled to 0°C, and a tetrahydrofuran solution (40 mL) of 11f (22.3 g, 77.1 mmol) was added dropwise, followed by a tetrahydrofuran solution (50 mL) of 4-toluenesulfonyl chloride (14.6 g, 77.1 mmol). The reaction mixture was stirred at 0°C for 6 hours. The temperature was lowered to -20°C, and a saturated ammonium chloride solution (10 mL) was slowly added, and the mixture was stirred vigorously until no gas was generated. The mixture was extracted with ethyl acetate (300 mL×3). The organic phases were combined, washed with saturated brine (150 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 1 / 1) to obtain the target product 11g (12.1 g, oily substance) with a yield of 58%.

[0430] MS m / z (ESI): 172 [M+1-100]

[0431] Step 7

[0432] ( S )-3-methyl-4-carbonyl-2-oxa-8-azaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (11h)

[0433] Slowly add Dess-Martin periodinane (37.8 g, 89.2 mmol) to a solution of 11g (12.1 g, 44.6 mmol) in dichloromethane (140 mL) at 0°C, and then stir at room temperature for 16 hours. Add saturated sodium bicarbonate solution until the solution is neutral, and extract with dichloromethane (150 mL×3). The organic phases are combined and washed with saturated brine (80 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue is purified by silica gel column chromatography (ethyl acetate / petroleum ether = 3 / 7) to obtain the target product 11h (8.36 g, oily substance) with a yield of 70%.

[0434] MS m / z (ESI): 170 [M+1-100]

[0435] Step 8

[0436] ( S )-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-one (11i)

[0437] 11h (1.2 g, 4.46 mmol) was dissolved in a solution of hydrogen chloride in dioxane (4 M, 20 mL) and stirred at room temperature for 2 hours. The solvent was removed under reduced pressure to obtain the target product 11i (hydrochloride, 800 mg, solid) with a yield of 87%.

[0438] MS m / z (ESI): 170 [M+1]

[0439] Step 9

[0440] ( S )-8-(5-iodo-1-methyl-6-carbonyl-1,6-dihydropyrimidin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-one (11j)

[0441] To 2-chloro-5-iodo-3-methylpyrimidine-4(3 H )-ketone 1c (130 mg, 0.481 mmol) in acetonitrile (20 mL) was added with 11i (hydrochloride, 108 mg, 0.529 mmol) and potassium carbonate (199 mg, 1.44 mmol), heated to 80°C and stirred for 16 hours. Cooled to room temperature, water (50 mL) was added, and extracted with ethyl acetate (30 mL×3). The organic phases were combined and washed with saturated brine (20 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 1.3 / 1) to obtain the target product 11j (180 mg, solid) with a yield of 92%.

[0442] MS m / z (ESI): 404 [M+1]

[0443] Step 10

[0444] ( R )-N-((3 S ,4 S )-8-(5-iodo-1-methyl-6-carbonyl-1,6-dihydropyrimidin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decane-4-yl)-2-methylpropane-2-sulfenamide (11k)

[0445] To a solution of 11j (180 mg, 0.446 mmol) in tetrahydrofuran (2 mL) was added ( R)-2-methylpropane-2-sulfenamide (108 mg, 0.892 mmol) and tetraethyl titanate (813 mg, 3.59 mmol), heated to 90°C, and stirred for 16 hours. Cooled to 0°C, methanol (5 mL) and lithium borohydride tetrahydrofuran solution (2.0 M, 0.23 mL, 0.46 mmol) were added and stirred for 1 hour. Quenched with saturated ammonium chloride solution and filtered. The filtrate was extracted with ethyl acetate (25 mL×3). The organic phases were combined and washed with saturated brine (20 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (methanol / dichloromethane = 1 / 16) to obtain the target product 11k (90 mg, solid), with a yield of 39%.

[0446] MS m / z (ESI): 509 [M+1]

[0447] Step 11

[0448] ( R )-N-((3 S ,4 S )-8-(5-((4-chloro-2-methyl-2H-indazol-5-yl)thio)-1-methyl-6-carbonyl-1,6-dihydropyrimidin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decane-4-yl)-2-methylpropane-2-sulfinamide (11l)

[0449] 7e (60 mg, 0.27 mmol), DIPEA (60.7 mg, 0.471 mmol), XantPhos (14.2 mg, 0.023 mmol) and Pd2(dba)3 (14.4 mg, 0.0157 mmol) were added to a solution of 11k (80 mg, 0.157 mmol) in dioxane (5 mL), and the mixture was heated to 80°C under nitrogen protection for 4 hours. The mixture was cooled to room temperature and desolventized under reduced pressure. The residue was purified by silica gel column chromatography (methanol / dichloromethane = 1 / 13) to obtain the target product 11l (55 mg, solid) with a yield of 60%.

[0450] MS m / z (ESI): 579 [M+1]

[0451] Step 12

[0452] 2-((3 S ,4 S )-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decane-8-yl)-5-((4-chloro-2-methyl-2H-indazol-5-yl)thio)-3-methylpyrimidine-4(3 H)-Ketoformate (11)

[0453] 11l (55 mg, 0.095 mmol) was dissolved in methanol (5 mL), and a solution of hydrogen chloride in dioxane (4.0 M, 5 mL) was added, and the mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure, and the residue was purified by reverse phase preparative high performance liquid chromatography to obtain the target product 11 (16.8 mg, solid) with a yield of 37%.

[0454] MS m / z (ESI): 475 [M+1]

[0455] 1 H NMR (400 MHz, DMSO- d 6 ) δ 8.43 (s, 1H), 8.21 (s, 1H), 7.91 (s, 1H),7.51 (d, J = 9.1 Hz, 1H), 6.99 (d, J = 9.0 Hz, 1H), 4.18 (s, 3H), 4.11 – 4.05(m, 1H), 3.68 (d, J = 8.6 Hz, 1H), 3.51 (d, J = 8.6 Hz, 1H), 3.47 – 3.43 (m,2H), 3.41 (s, 3H), 3.20 – 3.03 (m, 2H), 3.00 (d, J = 4.9 Hz, 1H), 1.89 – 1.82(m, 1H), 1.77 – 1.72 (m, 1H), 1.62 – 1.53 (m, 2H), 1.11 (d, J = 6.4 Hz, 3H).

[0456] Example 12

[0457] 2-((3 S ,4 S )-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-5-((4-chloropyrazolo[1,5- a ]pyridin-5-yl)thio)-3-methylpyrimidine-4(3 H )-Ketoformate

[0458]

[0459] first step

[0460] (R )-N-((3 S ,4 S )-8-(5-((4-chloropyrazolo[1,5- a ]pyridin-5-yl)thio)-1-methyl-6-carbonyl-1,6-dihydropyrimidin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decane-4-yl)-2-methylpropane-2-sulfenamide (12a)

[0461] 11k (140 mg, 0.28 mmol) was dissolved in dioxane (16 mL), and 8h (60 mg, 0.29 mmol), DIPEA (71.5 mg, 0.56 mmol), Pd2(dba)3 (25.3 mg, 0.03 mmol) and XantPhos (32 mg, 0.06 mmol) were added. The reaction mixture was heated to 100°C under nitrogen protection and reacted for 16 hours. The mixture was cooled to room temperature and desolventized under reduced pressure. The residue was purified by silica gel column chromatography (methanol / dichloromethane = 1 / 25) to obtain the target product 12a (100 mg, solid) with a yield of 64%.

[0462] MS m / z (ESI): 565 [M+1]

[0463] Step 2

[0464] 2-((3 S ,4 S )-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-5-((4-chloropyrazolo[1,5- a ]pyridin-5-yl)thio)-3-methylpyrimidine-4(3 H )-Ketoformate (12)

[0465] 12a (86 mg, 0.166 mmol) was dissolved in methanol (5 mL), and a solution of hydrogen chloride in dioxane (4.0 M, 1 mL) was added, and the mixture was stirred at room temperature for 1 hour. The mixture was concentrated under reduced pressure, and the residue was purified by reverse phase preparative HPLC to obtain the target product 12 (16.8 mg, solid) with a yield of 19%.

[0466] MS m / z (ESI): 461 [M+1]

[0467] 1 H NMR (400 MHz, DMSO- d 6 ) δ 8.51 (d, J= 7.3 Hz, 1H), 8.26 (s, 1H), 8.17 (s, 1H), 8.04 (d, J = 2.1 Hz, 1H), 6.59 (d, J = 1.5 Hz, 1H), 6.48 (d, J = 7.3 Hz, 1H), 4.14 – 4.07 (m, 1H), 3.72 (d, J = 8.7 Hz, 1H), 3.57 – 3.51 (m,3H), 3.39 (s, 3H), 3.21 – 3.07 (m, 2H), 3.06 (d, J = 4.9 Hz, 1H), 1.88 – 1.75(m, 2H), 1.66 – 1.55 (m, 2H), 1.12 (d, J = 6.4 Hz, 3H).

[0468] Embodiment 13

[0469] ( R )-5-((1H-indazol-4-yl)thio)-6-amino-2-(1-amino-8-azaspiro[4.5]decane-8-yl)-3-methylpyrimidine-4(3 H )-Ketoformate

[0470]

[0471] first step

[0472] 6-amino-5-bromo-3-methylpyrimidine-2,4(1 H ,3 H )-dione (13b)

[0473] 6-amino-3-methylpyrimidine-2,4(1 H ,3 H )-dione (1.0 g, 7.08 mmol) was dissolved in DMF (6 mL), and NBS (1.38 g, 7.8 mmol) was added. The mixture was stirred at room temperature for 16 hours. Water (20 mL) was added for dilution and the mixture was filtered. The filter cake was washed with water (5 mL×2) and dried under vacuum to obtain the target product 13b (1.0 g, solid) with a yield of 64%.

[0474] MS m / z (ESI): 220 [M+1]

[0475] Step 2

[0476] 8-(4-Amino-5-bromo-1-methyl-6-carbonyl-1,6-dihydropyrimidin-2-yl)-8-azaspiro[4.5]decan-1-one (13c)

[0477] 13b (200 mg, 0.91 mmol) was dissolved in DMF (5 mL), and 8-azaspiro[4.5]decane-1-one (167 mg, 1.1 mmol), Carter condensation agent (1.2 g, 2.73 mmol) and DBU (692 mg, 4.55 mmol) were added, and stirred at room temperature overnight. Ethyl acetate (50 mL) was added to dilute, and the mixture was washed with water (10 mL×3) and saturated brine (10 mL). The organic phase was desolvated under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 7 / 3) to obtain the target product 13c (170 mg, oily substance) with a yield of 53%.

[0478] MS m / z (ESI): 355 [M+1]

[0479] Step 3

[0480] ( R )-N-(( R )-8-(4-amino-5-bromo-1-methyl-6-carbonyl-1,6-dihydropyrimidin-2-yl)-8-azaspiro[4.5]decane-1-yl)-2-methylpropane-2-sulfinamide (13d)

[0481] To a solution of 13c (160 mg, 0.45 mmol) in tetrahydrofuran (15 mL) was added ( R )-2-methylpropane-2-sulfenamide (110 mg, 0.9 mmol) and tetraethyl titanate (411 mg, 1.8 mmol), heated to 90°C, and stirred overnight. Cooled to room temperature, methanol (5 mL) and lithium borohydride tetrahydrofuran solution (2.0 M, 0.5 mL) were added, and stirred for 1 hour. Concentrated under reduced pressure, the residue was purified by silica gel column chromatography (methanol / dichloromethane = 1 / 20) to obtain the target product 13d (200 mg, oily substance), with a yield of 96%.

[0482] MS m / z (ESI): 460 [M+1]

[0483] Step 4

[0484] ( R )-N-(( R)-8-(5-((1H-indazol-4-yl)thio)-4-amino-1-methyl-6-carbonyl-1,6-dihydropyrimidin-2-yl)-8-azaspiro[4.5]decane-1-yl)-2-methylpropane-2-sulfenamide (13e)

[0485] 13d (100 mg, 0.22 mmol) was dissolved in dioxane (10 mL), and sodium 1H-indazole-4-thiolate 3c (75 mg, 0.44 mmol), DIPEA (84 mg, 0.66 mmol), Pd2(dba)3 (40 mg, 0.044 mmol) and XantPhos (51 mg, 0.088 mmol) were added. The reaction mixture was heated to 100°C under nitrogen protection and stirred overnight. The temperature was cooled to room temperature, and the solvent was removed under reduced pressure. The residue was purified by thin layer silica gel preparative chromatography to obtain the target product 13e (30 mg, solid) with a yield of 26%.

[0486] MS m / z (ESI): 530 [M+1]

[0487] Step 5

[0488] ( R )-5-((1H-indazol-4-yl)thio)-6-amino-2-(1-amino-8-azaspiro[4.5]decane-8-yl)-3-methylpyrimidine-4(3 H )-Ketoformate (13)

[0489] 13e (30 mg, 0.056 mmol) was dissolved in methanol (2 mL), and a solution of hydrogen chloride in dioxane (4.0 M, 2 mL) was added, and the mixture was stirred at room temperature for 1 hour. The mixture was concentrated under reduced pressure, and the residue was purified by reverse phase preparative HPLC to obtain the target product 13 (5 mg, solid) with a yield of 21%.

[0490] MS m / z (ESI): 426 [M+1]

[0491] 1 H NMR (400 MHz, CD3OD) δ 8.54 (s, 1H), 8.12 (s, 1H), 7.30 (d, J = 8.3Hz, 1H), 7.20 (t, J = 7.7 Hz, 1H), 6.78 (d, J = 7.0 Hz, 1H), 3.65 – 3.57 (m,2H), 3.42 (s, 3H), 3.27 – 3.25 (m, 1H), 3.09 (t,J = 12.3 Hz, 2H), 2.22 –2.17 (m, 1H), 1.85 – 1.72 (m, 7H), 1.54 (t, J = 13.5 Hz, 2H).

[0492] Embodiment 14

[0493] 6-amino-2-((3 S ,4 S )-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decane-8-yl)-5-((4-chloro-2-methyl-2H-indazol-5-yl)thio)-3-methylpyrimidine-4(3 H )-Ketoformate

[0494]

[0495] first step

[0496] ( S )-8-(4-amino-1-methyl-6-carbonyl-1,6-dihydropyrimidin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-one (14a)

[0497] 6-amino-3-methylpyrimidine-2,4(1 H ,3 H )-dione 13a (400 mg, 2.8 mmol) was dissolved in DMF (25 mL), and 11i (576 mg, 2.8 mmol), Carter condensation agent (6.18 g, 14 mmol) and DBU (1.29 g, 8.5 mmol) were added and stirred at room temperature overnight. Water (60 mL) was added to quench the mixture, and the mixture was extracted with a mixed solvent of dichloromethane and methanol (10 / 1 v / v, 50 mL×3). The organic phases were combined and washed with saturated brine (20 mL×3), dried over anhydrous sodium sulfate, and desolventized under reduced pressure. The residue was purified by silica gel column chromatography (methanol / dichloromethane = 1 / 9) to obtain the target product 14a (700 mg, oily substance) with a yield of 84%.

[0498] MS m / z (ESI): 293 [M+1]

[0499] Step 2

[0500] ( R )-N-((3 S ,4 S)-8-(4-amino-1-methyl-6-carbonyl-1,6-dihydropyrimidin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decane-4-yl)-2-methylpropane-2-sulfenamide (14b)

[0501] To a solution of 14a (700 mg, 2.3 mmol) in tetrahydrofuran (30 mL) was added ( R )-2-methylpropane-2-sulfenamide (557 mg, 4.6 mmol) and tetraethyl titanate (2.06 g, 9.2 mmol), heated to 90°C, and stirred for 16 hours. Cooled to 0°C, methanol (10 mL) and lithium borohydride tetrahydrofuran solution (2.0 M, 1.35 mL) were added and stirred for 1 hour. Ammonium chloride solution was added to quench, filtered, and the filtrate was extracted with ethyl acetate (25 mL×3). The organic phases were combined and dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (methanol / dichloromethane = 1 / 10) to obtain the target product 14b (80 mg, solid), with a yield of 8.7%.

[0502] MS m / z (ESI): 398 [M+1]

[0503] Step 3

[0504] ( R )-N-((3 S ,4 S )-8-(4-amino-5-iodo-1-methyl-6-carbonyl-1,6-dihydropyrimidin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decane-4-yl)-2-methylpropane-2-sulfenamide (14c)

[0505] 14b (80 mg, 0.2 mmol) was dissolved in acetonitrile (10 mL), and NIS (54.2 mg, 0.24 mmol) was added and stirred at room temperature for 1.5 hours. The solvent was removed under reduced pressure, and the residue was purified by silica gel column chromatography (methanol / dichloromethane = 1 / 12) to obtain the target product 14c (100 mg, solid) with a yield of 95%.

[0506] MS m / z (ESI): 524 [M+1]

[0507] Step 4

[0508] ( R )-N-((3 S ,4 S)-8-(4-amino-5-((4-chloro-2-methyl-2H-indazol-5-yl)thio)-1-methyl-6-carbonyl-1,6-dihydropyrimidin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decane-4-yl)-2-methylpropane-2-sulfenamide (14d)

[0509] 14c (50 mg, 0.095 mmol) was dissolved in dioxane (6 mL), and 7e (22.8 mg, 0.114 mmol), DIPEA (36.7 mg, 0.285 mmol), Pd2(dba)3 (8.6 mg, 0.0095 mmol) and XantPhos (8.2 mg, 0.014 mmol) were added. The reaction mixture was heated to 80°C under nitrogen protection and stirred for 16 hours. The mixture was cooled to room temperature, diluted with ethyl acetate (100 mL), and washed with saturated brine. The organic phase was dried over anhydrous sodium sulfate, desolvated under reduced pressure, and the residue was purified by silica gel column chromatography (methanol / dichloromethane = 1 / 14) to obtain the target product 14d (46 mg, solid) with a yield of 82%.

[0510] MS m / z (ESI): 594 [M+1]

[0511] Step 5

[0512] 6-amino-2-((3 S ,4 S )-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decane-8-yl)-5-((4-chloro-2-methyl-2H-indazol-5-yl)thio)-3-methylpyrimidine-4(3 H )-Ketoformate (14)

[0513] 14d (46 mg, 0.077 mmol) was dissolved in methanol (5 mL), and a solution of hydrogen chloride in dioxane (4.0 M, 5 mL) was added, and the mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure, and the residue was purified by reverse phase preparative high performance liquid chromatography to obtain the target product 14 (6.5 mg, solid) with a yield of 17%.

[0514] MS m / z (ESI): 490 [M+1]

[0515] 1 H NMR (400 MHz, DMSO- d 6 ) δ 8.33 (s, 1H), 8.20 (s, 1H), 7.46 (d, J =9.0 Hz, 1H), 6.76 (d,J = 9.0 Hz, 1H), 4.15 (s, 3H), 4.10 – 4.06 (m, 1H), 3.69 (d, J = 8.5 Hz, 1H), 3.52 (d, J = 8.5 Hz, 1H), 3.39 – 3.37 (m, 2H), 3.29(s, 3H), 3.11 – 2.97 (m, 3H), 1.85 – 1.71 (m, 2H), 1.63 – 1.54 (m, 2H), 1.11(d, J = 6.4 Hz, 3H).

[0516] Embodiment 15

[0517] 6-amino-2-((3 S ,4 S )-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-5-((4-chloropyrazolo[1,5- a ]pyridin-5-yl)thio)-3-methylpyrimidine-4(3 H )-Ketoformate

[0518]

[0519] first step

[0520] ( R )-N-((3 S ,4 S )-8-(4-amino-5-((4-chloropyrazolo[1,5- a ]pyridin-5-yl)thio)-1-methyl-6-carbonyl-1,6-dihydropyrimidin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decane-4-yl)-2-methylpropane-2-sulfenamide (15a)

[0521] 14c (40 mg, 0.076 mmol) was dissolved in dioxane (4 mL), and 8h (31.3 mg, 0.152 mmol), DIPEA (29.4 mg, 0.228 mmol), Pd2(dba)3 (6.9 mg, 0.0076 mmol) and XantPhos (6.5 mg, 0.0114 mmol) were added. The reaction mixture was heated to 80°C under nitrogen protection and stirred for 16 hours. The mixture was cooled to room temperature, diluted with ethyl acetate (100 mL), and washed with saturated brine. The organic phase was dried over anhydrous sodium sulfate, desolvated under reduced pressure, and the residue was purified by silica gel column chromatography (methanol / dichloromethane = 1 / 17) to obtain the target product 15a (35 mg, solid) with a yield of 79%.

[0522] MS m / z (ESI): 580 [M+1]

[0523] Step 2

[0524] 6-amino-2-((3 S ,4 S )-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-5-((4-chloropyrazolo[1,5- a ]pyridin-5-yl)thio)-3-methylpyrimidine-4(3 H )-Ketoformate (15)

[0525] 15a (35 mg, 0.06 mmol) was dissolved in methanol (5 mL), and a solution of hydrogen chloride in dioxane (4.0 M, 5 mL) was added, and the mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure, and the residue was purified by reverse phase preparative HPLC to obtain the target product 15 (4.2 mg, solid) with a yield of 15%.

[0526] MS m / z (ESI): 476 [M+1]

[0527] 1 H NMR (400 MHz, CD3OD) δ 8.52 (s, 1H), 8.28 (d, J = 8.0 Hz, 1H), 7.93(d, J = 2.3 Hz, 1H), 6.55 (d, J = 1.7 Hz, 1H), 6.45 (d, J = 7.3 Hz, 1H), 4.28– 4.23 (m, 1H), 3.89 (d, J = 8.9 Hz, 1H), 3.77 (d,J = 8.9 Hz, 1H), 3.63 –3.56 (m, 2H), 3.43 (s, 3H), 3.24 – 3.02 (m, 3H), 1.97 – 1.90 (m, 2H), 1.79 –1.69 (m, 2H), 1.26 (d, J = 6.5 Hz, 3H).

[0528] Example 16

[0529] 2-(4-(Aminomethyl)-4-methylpiperidin-1-yl)-5-((4-chloro-2-methyl-2H-indazol-5-yl)thio)-3-methylpyrimidine-4(3 H )-Ketoformate

[0530]

[0531] first step

[0532] Tert-butyl ((1-benzyl-4-methylpiperidin-4-yl)methyl)carbamate (16b)

[0533] The reaction mixture of 1-benzyl-4-methylpiperidine-4-carbonitrile 16a (2 g, 9.3 mmol), di-tert-butyl dicarbonate (6.1 g, 27.9 mmol), nickel chloride hexahydrate (2.2 g, 9.3 mmol) and methanol (50 mL) was stirred at room temperature for 15 minutes. The mixture was cooled to 0°C, sodium borohydride (1.77 g, 46.5 mmol) was added, the mixture was warmed to room temperature and stirred for 8 hours. The solvent was removed under reduced pressure, and the residue was suspended in dichloromethane and filtered. The filtrate was removed under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 1 / 1) to obtain the target product 16b (1.2 g, oily substance) with a yield of 40%.

[0534] MS m / z (ESI): 319 [M+1]

[0535] Step 2

[0536] Tert-butyl ((4-methylpiperidin-4-yl)methyl)carbamate (16c)

[0537] 16b (1.2 g, 3.77 mmol) was dissolved in methanol (20 mL), palladium carbon (10%, water content 55%, 1.2 g) was added, and stirred at room temperature for 3 hours under hydrogen atmosphere. The mixture was filtered, the filter cake was washed with methanol, and the filtrate was concentrated to obtain the target product 16c (810 mg, oily substance) with a yield of 94%.

[0538] MS m / z (ESI): 229 [M+1]

[0539] Step 3

[0540] Tert-butyl ((1-(5-iodo-1-methyl-6-carbonyl-1,6-dihydropyrimidin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (16d)

[0541] 1c (150 mg, 0.55 mmol) was added to a mixture of 16c (150 mg, 0.66 mmol), potassium carbonate (228 mg, 1.65 mmol) and acetonitrile (10 mL), heated to 90°C and stirred for 16 hours. The mixture was concentrated under reduced pressure and the residue was purified by silica gel column chromatography (methanol / dichloromethane = 1 / 9) to give the target product 16d (150 mg, solid) in a yield of 58%.

[0542] MS m / z (ESI): 463 [M+1]

[0543] Step 4

[0544] Tert-butyl ((1-(5-((4-chloro-2-methyl-2H-indazol-5-yl)thio)-1-methyl-6-carbonyl-1,6-dihydropyrimidin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (16e)

[0545] The reaction mixture 16d (150 mg, 0.32 mmol), 7e (86 mg, 0.39 mmol), DIPEA (124 mg, 0.96 mmol), Pd2(dba)3 (29 mg, 0.032 mmol), XantPhos (18.5 mg, 0.032 mmol) and dioxane (4 mL) were heated to 90°C under nitrogen protection and stirred for 3 hours. The solvent was removed under reduced pressure and the residue was purified by silica gel column chromatography (methanol / dichloromethane = 1 / 9) to obtain the target product 16e (130 mg, solid) with a yield of 75%.

[0546] MS m / z (ESI): 533 [M+1]

[0547] Step 5

[0548] 2-(4-(Aminomethyl)-4-methylpiperidin-1-yl)-5-((4-chloro-2-methyl-2H-indazol-5-yl)thio)-3-methylpyrimidine-4(3 H )-Ketoformate (16)

[0549] 16e (130 mg, 0.24 mmol) was dissolved in dichloromethane (5 mL), trifluoroacetic acid (1 mL) was added, and the mixture was stirred at room temperature for 1 hour. The mixture was concentrated under reduced pressure, and the residue was purified by reverse phase preparative HPLC to obtain the target product 16 (32.4 mg, solid) with a yield of 31%.

[0550] MS m / z (ESI): 433 [M+1]

[0551] 1 H NMR (400 MHz, DMSO- d 6 ) δ 8.43 (s, 1H), 8.37 (s, 1H), 7.91 (s, 1H),7.51 (d, J = 9.0 Hz, 1H), 6.99 (d, J = 9.0 Hz, 1H), 4.18 (s, 3H), 3.40 – 3.36(m, 5H), 3.17 (t, J = 10.2 Hz, 2H), 2.65 (s, 2H), 1.63 – 1.57 (m, 2H), 1.42(d, J = 11.1 Hz, 2H), 1.01 (s, 3H).

[0552] Embodiment 17

[0553] 2-((3 S ,4 S )-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-3-methyl-5-((1-methyl-1H-pyrrolo[2,3- b ]pyridin-4-yl)thio)pyrimidine-4(3 H )-Ketoformate

[0554]

[0555] first step

[0556] ( R )-2-methyl-N-((3 S ,4 S )-3-methyl-2-oxa-8-azaspiro[4.5]decane-4-yl)propane-2-sulfenamide (17b)

[0557] (3 S ,4 S )-4-((( Rtert-Butyl ((tert-butylsulfinyl)amino)-3-methyl-2-oxa-8-azaspiro[4.5]decane-8-carboxylate 17a (300 mg, 0.8 mmol) was dissolved in dichloromethane (5 mL), trifluoroacetic acid (1 mL) was added, and stirred at room temperature for 1 hour. The mixture was concentrated under reduced pressure to obtain the target product 17b (350 mg, crude product). The product was used directly in the next step without purification.

[0558] MS m / z (ESI): 275 [M+1]

[0559] Step 2

[0560] ( R )-N-((3 S ,4 S )-8-(5-iodo-1-methyl-6-carbonyl-1,6-dihydropyrimidin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decane-4-yl)-2-methylpropane-2-sulfenamide (17c)

[0561] The reaction mixture 1c (216 mg, 0.8 mmol), 17b (crude product, 350 mg, 0.8 mmol), potassium carbonate (562 mg, 4.05 mmol) and acetonitrile (10 mL) were heated to 80°C and stirred for 16 hours. The mixture was concentrated under reduced pressure and the residue was purified by silica gel column chromatography (methanol / dichloromethane = 1 / 9) to give the target product 17c (360 mg, solid) in a yield of 89%.

[0562] MS m / z (ESI): 509 [M+1]

[0563] Step 3

[0564] ( R )-2-methyl-N-((3 S ,4 S )-3-methyl-8-(1-methyl-5-((1-methyl-1H-pyrrolo[2,3- b ]pyridin-4-yl)thio)-6-carbonyl-1,6-dihydropyrimidin-2-yl)-2-oxa-8-azaspiro[4.5]decane-4-yl)propane-2-sulfenamide (17d)

[0565] The reaction mixture 17c (180 mg, 0.35 mmol), 2c (70 mg, 0.42 mmol), DIPEA (135 mg, 1.05 mmol), Pd2(dba)3 (32 mg, 0.035 mmol), XantPhos (20 mg, 0.035 mmol) and dioxane (5 mL) were heated to 90°C under nitrogen protection and stirred for 2 hours. The mixture was cooled to room temperature and desolventized under reduced pressure. The residue was purified by silica gel column chromatography (methanol / dichloromethane = 1 / 9) to obtain the target product 17d (160 mg, solid) with a yield of 83%.

[0566] MS m / z (ESI): 545 [M+1]

[0567] Step 4

[0568] 2-((3 S ,4 S )-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-3-methyl-5-((1-methyl-1H-pyrrolo[2,3- b ]pyridin-4-yl)thio)pyrimidine-4(3 H )-Ketoformate (17)

[0569] 17d (160 mg, 0.29 mmol) was dissolved in methanol (4 mL), and a solution of hydrogen chloride in dioxane (4.0 M, 2 mL) was added, and the mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure, and the residue was purified by reverse phase preparative HPLC to obtain the target product 17 (36 mg, solid) with a yield of 28%.

[0570] MS m / z (ESI): 441 [M+1]

[0571] 1 H NMR (400 MHz, DMSO- d 6 ) δ 8.22 (s, 1H), 8.14 (s, 1H), 8.03 (d, J =5.1 Hz, 1H), 7.51 (d, J = 3.5 Hz, 1H), 6.58 (d, J = 5.1 Hz, 1H), 6.44 (d, J =3.5 Hz, 1H), 4.12 – 4.04 (m, 1H), 3.80 (s, 3H), 3.70 (d, J= 8.6 Hz, 1H), 3.53 – 3.48 (m, 3H), 3.40 (s, 3H), 3.19 – 3.08 (m, 2H), 3.02 (d, J = 4.9 Hz,1H), 1.88 – 1.84 (m, 2H), 1.64 – 1.54 (m, 2H), 1.11 (d, J = 6.4 Hz, 3H).

[0572] Embodiment 18

[0573] 5-((1H-pyrrolo[2,3- b ]pyridin-4-yl)thio)-2-((3 S ,4 S )-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decane-8-yl)-3-methylpyrimidine-4(3 H )-Ketoformate

[0574]

[0575] first step

[0576] 1H-pyrrolo[2,3- b ]Pyridine-4-thiol (18a)

[0577] Potassium tert-butoxide (224 mg, 2 mmol) was added to a solution of 1 g (250 mg, 1 mmol) in tetrahydrofuran (5 mL) and stirred at room temperature for 2 hours. The mixture was quenched with water and washed with ethyl acetate. The aqueous phase was acidified with hydrochloric acid (6 N) to pH 4 and extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, filtered to remove the desiccant, and desolventized under reduced pressure to obtain the target product 18a (100 mg, solid) with a yield of 67%.

[0578] MS m / z (ESI): 151 [M+1]

[0579] Step 2

[0580] ( R )-N-((3 S ,4 S )-8-(5-((1H-pyrrolo[2,3- b ]pyridin-4-yl)thio)-1-methyl-6-carbonyl-1,6-dihydropyrimidin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decane-4-yl)-2-methylpropane-2-sulfenamide (18b)

[0581] The reaction mixture 17c (180 mg, 0.35 mmol), 18a (63 mg, 0.42 mmol), DIPEA (135 mg, 1.05 mmol), Pd2(dba)3 (32 mg, 0.035 mmol), XantPhos (20 mg, 0.035 mmol) and dioxane (5 mL) were heated to 90°C under nitrogen protection and stirred for 2 hours. The mixture was cooled to room temperature and desolventized under reduced pressure. The residue was purified by silica gel column chromatography (methanol / dichloromethane = 1 / 9) to obtain the target product 18b (150 mg, solid) with a yield of 80%.

[0582] MS m / z (ESI): 531 [M+1]

[0583] Step 3

[0584] 5-((1H-pyrrolo[2,3- b ]pyridin-4-yl)thio)-2-((3 S ,4 S )-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decane-8-yl)-3-methylpyrimidine-4(3 H )-Ketoformate (18)

[0585] 18b (150 mg, 0.28 mmol) was dissolved in methanol (4 mL), and a solution of hydrogen chloride in dioxane (4.0 M, 2 mL) was added, and the mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure, and the residue was purified by reverse phase preparative HPLC to obtain the target product 18 (37.9 mg, solid) with a yield of 31%.

[0586] MS m / z (ESI): 427 [M+1]

[0587] 1 H NMR (400 MHz, DMSO- d 6 ) δ 11.73 (s, 1H), 8.25 (s, 1H), 8.14 (s, 1H), 7.99 (d, J = 5.1 Hz, 1H), 7.47 – 7.45 (m, 1H), 6.54 (d, J = 5.1 Hz, 1H), 6.42(d, J = 2.3 Hz, 1H), 4.15 – 4.05 (m, 1H), 3.71 (d, J= 8.5 Hz, 1H), 3.52 –3.48 (m, 3H), 3.41 (s, 3H), 3.24 – 3.05 (m, 3H), 1.89 – 1.75 (m, 2H), 1.65 –1.55 (m, 2H), 1.12 (d, J = 6.2 Hz, 3H).

[0588] Embodiment 19

[0589] 2-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decane-8-yl)-5-((4-chloro-2-methyl-2H-indazol-5-yl)thio)-3-(methyl-d3)pyrimidin-4(3H)-one

[0590]

[0591] first step

[0592] (3S,4S)-3-Methyl-2-oxa-8-azaspiro[4.5]decan-4-amine (19b)

[0593] A solution of (3S,4S)-4-(((R)-tert-butylsulfinyl)amino)-3-methyl-2-oxa-8-azaspiro[4.5]decane-8-carboxylic acid tert-butyl ester 19a (95.0 g, 254 mmol) and hydrogen chloride in methanol (4 M, 1 L) were stirred at room temperature for 4 hours. The mixture was concentrated under reduced pressure, and the residue was dissolved in methanol (500 mL). A mixed solvent of petroleum ether and ethyl acetate (1 / 1, 4 L) was slowly added and stirred at room temperature for 2 hours. The target product 19b (64.1 g, solid, dihydrochloride) was obtained after filtration and drying of the filter cake. The yield was >100%.

[0594] MS m / z (ESI): 171[M+1]

[0595] 1H NMR (400 MHz, DMSO- d 6 ) δ 9.12 (brs, 2H), 8.33 (s, 3H), 4.25 – 4.19(m, 1H), 3.82 (d, J = 9.1 Hz, 1H), 3.64 (d, J= 9.1 Hz, 1H), 3.47 (brs, 1H), 3.27 – 3.24 (m, 1H), 3.19 – 3.16 (m, 1H), 2.94 – 2.85 (m, 2H), 2.03 – 1.85(m, 2H), 1.79 – 1.67 (m, 2H), 1.23 (d, J = 6.5 Hz, 3H).

[0596] Step 2

[0597] 2-Chloro-5-iodo-3-(methyl-d3)pyrimidin-4(3H)-one (19c)

[0598] DBU (3.85 g, 25.3 mmol) was added to a solution of 2-chloro-5-iodopyrimidin-4(3H)-one 1b (5.0 g, 19.5 mmol) in tetrahydrofuran (100 mL), the temperature was lowered to 0°C, deuterated iodomethane (3.39 g, 23.4 mmol) was added, the mixture was heated to 50°C, and stirred for 6 hours. The solvent was removed under reduced pressure, ethyl acetate (3.5 L) was added to the residue, and the mixture was washed with dilute hydrochloric acid (0.5 N, 1 L× 2) and brine (800 mL×2). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 1 / 19 to 1 / 4) to obtain the target product 19c (3.3 g, solid) with a yield of 62%.

[0599] MS m / z (ESI): 274[M+1]

[0600] Step 3

[0601] 2-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decane-8-yl)-5-iodo-3-(methyl-d3)pyrimidin-4(3H)-one (19d)

[0602] The reaction mixture 19c (1.8 g, 6.5 mmol), 19b (1.67 g, 6.9 mmol, dihydrochloride), cesium carbonate (9.5 g, 29.3 mmol) and acetonitrile (60 mL) were stirred at room temperature for 3 hours under nitrogen protection. The mixture was concentrated under reduced pressure and the residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 9 / 1) to obtain the target product 19d (2.4 g, solid) with a yield of 91%.

[0603] MS m / z (ESI): 408[M+1]

[0604] Step 4

[0605] 2-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decane-8-yl)-5-((4-chloro-2-methyl-2H-indazol-5-yl)thio)-3-(methyl-d3)pyrimidin-4(3H)-one (19)

[0606] The reaction mixture 7e (1.9 g, 9.0 mmol), 19d (2.3 g, 5.65 mmol), cuprous iodide (430 mg, 2.26 mmol), 1,10-phenanthroline (814 mg, 4.52 mmol), potassium phosphate (2.3 g, 11.3 mmol) and dioxane (50 mL) were heated to 100°C under nitrogen protection and stirred overnight. The temperature was cooled to room temperature and desolventized under reduced pressure. The residue was purified by silica gel column chromatography (methanol / dichloromethane = 1 / 9), and the obtained crude product (2.5 g) was added with acetonitrile (25 mL), stirred at 90°C for 3 hours, and then stirred at room temperature for 16 hours. Filtered, the filter cake was vacuum dried at 40°C for 4 hours to obtain the target product 19 (1.47 g, solid), with a yield of 54%.

[0607] MS m / z (ESI): 478[M+1]

[0608] 1H NMR (400 MHz, DMSO- d 6 ) δ 8.42 (s, 1H), 7.91 (s, 1H), 7.50 (dd, J =9.0, 0.9 Hz, 1H), 6.98 (d, J = 9.0 Hz, 1H), 4.17 (s, 3H), 4.05 – 4.02 (m,1H), 3.64 (d, J = 8.5 Hz, 1H), 3.47 (d, J = 8.5 Hz, 1H), 3.48 – 3.37 (m, 2H), 3.15 – 3.05 (m, 2H), 2.90 (d, J = 5.1 Hz, 1H), 1.86 – 1.80 (m, 1H), 1.74 –1.68 (m, 1H), 1.59 – 1.50 (m, 2H), 1.38 (brs, 2H), 1.07 (d, J = 6.4 Hz, 3H).

[0609] Biological experiments

[0610] Activity inhibition test of SHP2

[0611] Rapid fluorometric assay to evaluate the effects of compounds of the invention on SHP2 activity using the surrogate substrate DiFMUP

[0612] The experimental method is outlined as follows:

[0613] The recombinant human SHP2 full-length protein was expressed and purified by the protein purification and identification platform of Tsinghua University. The dual-phosphorylated peptide (H2N-LN(pY)IDLDLV-(dPEG8)LST(pY)ASINFQK-amide) was synthesized by GenScript Biotech Co., Ltd. The alternative substrate DiFMUP was purchased from Thermo Fisher Scientific (Cat. No. D6567). The reaction buffer contained the following components: 60 mM HEPES (pH 7.2), 75 mM NaCl, 75 mM KCl, 1 mM EDTA, 0.05% TWEEN 20, and 5 mM DTT.

[0614] The compounds were dissolved in DMSO (Sigma, Cat. No. D5879) to 100 μM, and then serially diluted 4-fold in DMSO to a minimum concentration of 6.1 nM. Each concentration point was further diluted 25-fold in reaction buffer.

[0615] Add 10 µL of compound solution and 10 µL of 0.25 nM SHP2 protein solution to a 384-well assay plate (Corning, catalog number 3575), mix well and incubate at room temperature for 30 minutes; add 10 µL of 0.5 µM dual phosphorylated peptide (dissolved in reaction buffer), mix well and incubate at room temperature for 30-60 minutes; then add 10 µL of 60 µM DiFMUP solution (dissolved in reaction buffer), mix well and leave at room temperature. After 30 minutes, the fluorescence signal was detected using an enzyme reader (EnSpire, Perkin Elmer) at an excitation wavelength of 340 nm / emission wavelength of 450 nm. The fluorescence signal value is positively correlated with the degree of dephosphorylation of the substrate, thus reflecting the catalytic activity of SHP2 phosphatase. In this experiment, the group without protein addition was used as the 100% inhibition group, and the group with protein but no compound addition was used as the 0% inhibition group. The compound inhibition curve was drawn using XLfit software and its inhibition IC was calculated. 50 , the experimental results are shown in the table.

[0616] Compound No. <![CDATA[IC 50 (nM)]]> 1. 23 2. 20 3. 20 4. 126 5. 801 6. 877 7. 12 8. 8.6 9. 8.8 10. 36 11. 4.0 12. 3.1 13. 8.7 14. 2.5 15. 2.7 16. 56 17. 14 18. 5.5 19. 11

[0617] The example compounds of the present invention have an inhibitory effect on the activity of SHP2, preferably IC 50 Less than 50 nM.

[0618] Determination of NCI-H358 cell proliferation inhibition

[0619] The luminescent cell viability test was used to evaluate the effect of the compounds of the present invention on the proliferation of NCI-H358 human non-small cell lung cancer cells.

[0620] The experimental method is outlined as follows:

[0621] The compound was dissolved in DMSO (Sigma, Catalog No. D5879) and diluted to 5 mM, then serially diluted 4-fold with DMSO to a minimum concentration of 0.31 μM, and each concentration point was further diluted 50-fold with RPMI 1640 medium (Thermo Fisher Scientific, Catalog No. 11995073). 50 Lower values ​​can reduce the starting concentration of the compound.

[0622] NCI-H358 (ATCC, catalog number CRL-5807) cells were cultured in RPMI 1640 complete medium [RPMI1640 medium contains 10% FBS (GBICO, catalog number 10099-141) and 100 units / mL penicillin-streptomycin mixture (Thermo Fisher Scientific, catalog number 15140122)]. Cells (15,000 cells / mL) were seeded in 90 μL complete medium in a 96-well plate, and 10 μL of compound solution was added to each well after overnight culture. The plates were cultured in a 37ºC, 5% CO2 incubator for 5 days, and then the plates were taken out and equilibrated to room temperature according to the instructions of the CellTilter-Glo (CTG) kit (Promega, catalog number G7572). 50 μL of CTG reagent was added to fully lyse the cells, and the cells were left at room temperature for 10 minutes before the luminescence signal was read using an ELISA reader (EnVision, Perkin Elmer). In the experiment, 10 μM positive control RMC-4550 was added as a negative control (100% inhibition), and 0.2% DMSO was added as a positive control (0% inhibition). XLfit software was used to draw the compound inhibition curve and calculate its inhibition IC 50 , the experimental results are shown in the following table.

[0623] Compound No. <![CDATA[IC 50 (nM)]]> 1 310 2 383 3 434 4 2359 7 575 8 309 9 1345 10 1763 11 165 12 264 14 108 15 342 17 540 18 335 19 222

[0624] The example compounds of the present invention have an inhibitory effect on cell proliferation, preferably IC 50 is less than 1000 nM.

[0625] Determination of hERG potassium channel blockade

[0626] The effects of the compounds of the present invention on potential arrhythmias were evaluated by measuring the blocking effect on hERG potassium channels.

[0627] The experimental method is outlined as follows:

[0628] Extracellular solution: 140 mM NaCl, 3.5 mM KCl, 1 mM MgCl2, 2 mM CaCl2, 10 mM D-glucose, 10 mM HEPES, 1.25 mM NaH2PO4, pH = 7.4.

[0629] Electrode solution: 20 mM KCl, 115 mM K-aspartate, 1 mM MgCl2, 5 mM EGTA, 10 mM HEPES, 2 mM Na2-ATP, pH = 7.2.

[0630] Compound solution: Dissolve the test compound in DMSO to prepare a 10 mM stock solution, dilute it to 3 mM with DMSO, and then dilute it to 3 µM with extracellular fluid for subsequent use.

[0631] Cell culture: HEK293 cells stably overexpressing hERG potassium channel (Creacell, Catalog No. A-0320) were cultured in DMEM medium (Gibco, Catalog No. 11995-065) containing 10% fetal bovine serum (Gibco, Catalog No. 1428478) and 0.8 mg / mL G418 (Amresco, Catalog No. E859-5G) at 37°C and 5% CO2. The old medium was removed and washed once with PBS (Gibco, Catalog No. 1009-141), and then 1 mL of TrypLE TM Express solution (Gibco, catalog number 12604021), incubate at 37°C for 30 seconds. When the cells detach from the bottom of the culture dish, add 5 mL of complete medium preheated at 37°C, transfer the cell suspension to a sterile centrifuge tube, and collect the cells after centrifugation at 1000 rpm for 5 minutes. Inoculate the cells in a 6 cm cell culture dish, with 2.5*10 cells in each cell culture dish. 5 (Final volume: 5 mL). Before the patch clamp test, 3*10 3 Cells were plated onto coverslips, cultured in 24-well plates (final volume: 500 µL), and assayed after 18 hours.

[0632] Voltage stimulation scheme for whole-cell patch clamp recording of whole-cell hERG potassium current: When the whole-cell seal is formed, the cell membrane voltage is clamped at -80 mV. The clamping voltage is depolarized from -80 mV to -50 mV and maintained for 0.5 seconds (as a leak current detection), then stepped to 30 mV and maintained for 2.5 seconds, and then quickly restored to -50 mV and maintained for 4 seconds to stimulate the tail current (Peak tail current) of the hERG channel. The hERG potassium current is recorded once every 10 seconds. The experimental data are collected by the EPC-10 amplifier (HEKA) and stored in the PatchMaster (HEKA v2x73) software.

[0633] Assay: Recording electrodes were made from capillary glass tubes (Sutter Instruments) using a microelectrode puller (Sutter Instruments). The coverslip containing cells was removed from a 24-well plate placed in an incubator and placed under an inverted microscope. The recording electrode was perfused with electrode solution, and then the microelectrode manipulator (Sutter Instruments) was manipulated to contact the recording electrode to the cell surface and apply negative pressure suction to form a GΩ seal; then fast capacitance compensation was performed; then negative pressure suction was continued until the cell membrane was broken and the whole-cell recording mode was formed. In the whole-cell recording mode, slow capacitance compensation was performed and membrane capacitance and series resistance were recorded without leakage compensation. When the hERG tail current recorded in the whole cell was stable for 3-5 minutes, 8 mL of extracellular solution without compound (blank control) and 8 mL of 3 µM test compound solution were passed through the recording bath by gravity perfusion to act on the cells for 5 minutes (or until the current stabilized). The current detected in the extracellular solution without compound for each cell was used as its own control. Two to three cells were tested independently. All electrophysiological experiments were performed at room temperature.

[0634] Data analysis: First, the current after the test compound is treated and the blank control current are standardized ( ), and then calculate the inhibition rate corresponding to this compound concentration (1 - ).

[0635] Compound No. Percent inhibition of hERG current at 3 µM concentration (n = 2) 7 10% 8 45% 9 65% 11 -0.5% 12 21% 14 -4% 15 1%

Claims

1. A compound of general formula (II) or a pharmaceutically acceptable salt thereof: in: Ring A is a benzene ring or a 6-membered heteroaromatic ring containing one nitrogen atom, and ring B is a 5-membered heteroaromatic ring containing one or two nitrogen atom fused to ring A, wherein the benzene ring and the heteroaromatic ring are optionally substituted by one or more selected from D, halogen, cyano, C 1-6 Alkyl, -OR a or NR a R b substituted by a substituent; R 1 H, D, cyano, C 1-2 Alkyl or -NR a R b ; R 2 H or C 1-2 Alkyl, wherein one or more hydrogen atoms of the alkyl group are optionally substituted by one or more D or fluorine atoms; R 4a and R 4b Each independently selected from C 1-6 Alkyl, the alkyl group is optionally substituted by one or more selected from D or -NR a R b Substituents; R 4a and R 4b Optional carbon atoms connected to form a C 3-7 A carbocyclic ring or a 4-7 membered heterocyclic ring containing one oxygen; R a and R b are each independently selected from H.

2. A compound of general formula (IV) or a pharmaceutically acceptable salt thereof: in: Ring A is a benzene ring or a 6-membered heteroaromatic ring containing one nitrogen atom, and ring B is a 5-membered heteroaromatic ring containing one or two nitrogen atom fused to ring A, wherein the benzene ring and the heteroaromatic ring are optionally substituted by one or more selected from D, halogen, cyano, C 1-6 Alkyl, -OR a or NR a R b substituted by a substituent; X is -O- or -CH2-; R 1 H, D, C 1-2 Alkyl or -NR a R b ; R 2 H or C 1-2 Alkyl, wherein one or more hydrogen atoms of the alkyl group are optionally substituted by one or more D or fluorine atoms; R 7 H, D or C 1-2 Alkyl, wherein one or more hydrogen atoms of the alkyl group are optionally substituted by one or more D or fluorine atoms; R a and R b Each is independently H.

3. A compound or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:

4. A pharmaceutical composition comprising the compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers and excipients.

5. A pharmaceutical composition comprising a compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof and at least one additional drug, wherein the at least one additional drug is selected from chemotherapeutic drugs, targeted drugs, DNA synthesis inhibitors, antibody drugs, antibody drug conjugates, anti-tumor drugs and immunosuppressants.

6. Use of the compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 4 or 5 in the preparation of a medicament for treating and / or preventing diseases associated with abnormal SHP2 activity.

7. The use according to claim 6, wherein the disease associated with abnormal SHP2 activity is selected from leukemia, Noonan syndrome, leopard spot syndrome, neuroblastoma, melanoma, lung cancer, breast cancer, esophageal cancer, colon cancer, head and neck cancer, and gastric cancer.

Images