Isoquinolinone and quinazolinone compounds and their compositions and uses

Abstract

The present invention belongs to the field of medicine, and specifically relates to isoquinolinone and quinazolinone compounds shown in formula (I) or (II), or stereoisomers, tautomers, deuterated substances, nitrogen oxides, solvates, or pharmaceutically acceptable salts thereof, and pharmaceutical compositions comprising the compounds, and the use of the compounds and pharmaceutical compositions thereof in the preparation of drugs for preventing, treating, treating, and / or alleviating PI3-kinase-mediated diseases, disorders, and / or conditions, or inhibiting PI3-kinase activity. The compounds provided by the present invention show excellent inhibitory activity and kinase selectivity to target kinases, and have good pharmacokinetic properties and higher bioavailability.

Description

Field of the Invention

[0001] The present invention belongs to the field of medicine, and specifically relates to a new class of isoquinolinone and quinazolinone compounds, pharmaceutically acceptable salts thereof and pharmaceutical compositions comprising the compounds, as well as the use of the compounds and the pharmaceutical compositions thereof in preparing drugs for preventing, treating, curing, and / or alleviating diseases, disorders and / or conditions associated with abnormal PI3-kinase. Background of the Invention

[0002] The phosphoinositide 3-kinase (PI3K) pathway is an intracellular signaling pathway with regulatory roles in cell survival, proliferation, and differentiation. The phosphoinositide 3-kinase (PI3K) family of enzymes are central regulators of growth, proliferation, migration, and metabolism in many cells and tissues. PI3Ks are lipid kinases that generate the lipid second messenger phosphatidylinositol-3,4,5-triphosphate (PIP3), which is used downstream of cell surface receptors to regulate growth, metabolism, survival, and differentiation. PIP3 is produced by four distinct class I PI3K catalytic isoforms, which are divided into two groups: class IA (p110α, p110β, and p110δ) and class IB (p110γ). All class I PI3Ks are constitutively associated with regulatory subunits, and the main difference between class IA and class IB PI3Ks is that they are associated with unique regulatory subunits. Class IA PI3Ks (PI3Kα, PI3Kβ, and PI3Kδ) are heterodimeric complexes composed of a catalytic subunit p110 (p110α, p110β, and p110δ, respectively) and a regulatory subunit p85 (e.g., p85α, p85β, p55δ, p55α, and p50α). These signaling responses are typically mediated through receptor tyrosine kinases (RTKs). Class IB PI3Kγ signals through G protein-coupled receptors (GPCRs) and consists of the catalytic subunit p110γ. In normal cells, the PI3K / mTOR pathway has regulatory roles in cell survival, proliferation, and differentiation. However, aberrant activation of this pathway has been implicated in a variety of human diseases, including cancer, immune deficiency, inflammation, and developmental disorders. Multiple inhibitors targeting key nodes within the PI3K pathway are in various stages of clinical development for the treatment of a variety of human diseases. ("Small-molecule inhibitors of the PI3K signaling network." Future MedChem. 2011, 3(5), 549-565).

[0003] The expression of PI3Kα and PI3Kβ is ubiquitous, while PI3Kδ and PI3Kγ are mainly expressed in leukocytes. The relatively restricted expression of PI3Kδ and PI3Kγ indicates the important roles of these two isoforms in the adaptive and innate immune systems (J. Med. Chem. 2012, 55(20), 8559–8581).

[0004] The PI3Kα isoform is involved in angiogenesis and glucose homeostasis. The PI3K / mTOR pathway is frequently dysregulated in cancer, usually due to activating mutations or amplifications of PIK3CA. Gain-of-function mutations in PIK3CA, the gene encoding the PI3K p110α catalytic subunit, are among the most common somatic alterations in solid tumors.

[0005] PI3Kδ is primarily expressed in cells of the hematopoietic lineage and is activated by cytokine receptors, antigen receptors, growth factor receptors, and co-stimulatory receptors. PI3Kδ is important in the development and activation of T and B cells. Blockade of the PI3Kδ signaling pathway increases genetic instability. Gain-of-function (GOF) mutations in PI3Kδ lead to a range of developmental and functional defects in B and T cells, compromising host defense. Loss-of-function (LOF) mutations lead to more severe B-cell lymphopenia and agammaglobulinemia, but do not cause T-cell senescence.

[0006] The class IB PI3K catalytic subunit, p110γ, is a master regulator of immune cell function and plays a key role in immune signaling. p110γ is a key player in inflammatory diseases and has been identified as a therapeutic target in cancer due to its immunomodulatory effects. PI3Kγ plays an important role in regulating myeloid (macrophages, mast cells, neutrophils) and lymphoid (T cells, B cells, and natural killer cells) derived immune cells. It regulates immune cell chemotaxis, cytokine release, and the production of reactive oxygen species. The ability of PI3Kγ to mediate a variety of immune cell functions is controlled by its activation downstream of numerous cell surface receptors, including G protein-coupled receptors (GPCRs), IgE / antigen receptors, receptor tyrosine kinases (RTKs), and Toll-like receptors (TLRs). In mouse models, genetic or pharmacological deletion of PI3Kγ is protective against a variety of inflammatory diseases, including cardiovascular disease, arthritis, lupus, asthma, lung inflammation and fibrosis, and metabolic syndrome. PI3Kγ is also a driver of pancreatic ductal adenocarcinoma progression through immunomodulatory effects. Targeting PI3Kγ in the immune system in combination with checkpoint inhibitors has shown promise in clinical cancer therapy studies (Eganelisib, Infinity Pharmaceuticals News Release, 2021). (Henau OD, Rausch M, Winkler D et al. Nature 539(7629), 443–447(2016); Kaneda MM, Messer KS, Ralainirina N et al. Nature 539(7629), 437–442(2016).)

[0007] Given its critical role in cancer and immunity, the PI3K pathway has been a focus of drug development research over the past two decades. In 2014, the PI3Kδ inhibitor idelalisib (Zydelig; Gilead Sciences) became the first PI3K inhibitor approved for specific B-cell malignancies. Subsequently, the pan-class I PI3K inhibitor copanlisib (Aliqopa; Bayer) was approved in 2017, and the dual PI3Kδ / PI3Kγ inhibitor duvelisib (Copiktra; Verastem, now Secura Bio) was approved for the same indication in 2018. The PI3Kα inhibitor alpelisib (Piqray; Novartis) is the first and only PI3Kα inhibitor approved by the FDA in May 2019 in combination with the estrogen receptor (ER) downregulator fulvestrant for the treatment of PIK3CA-mutated HER2 / ER+ metastatic advanced breast cancer.

[0008] It is necessary to provide new, good candidate drugs for PI3K inhibitors. Specifically, the preferred compounds should bind strongly to the PI3K receptor, show little affinity for other receptors, and show functional activity as agonists. The compounds should be fully absorbed by the gastrointestinal tract, be metabolically stable and have good pharmacokinetic properties. When targeting receptors in the central nervous system, they can freely cross the blood-brain barrier, and when selectively targeting receptors in the peripheral nervous system, they should not cross the blood-brain barrier. They should be non-toxic and show very few side effects. In addition, the ideal candidate drug should exist in a stable, non-hygroscopic and easily formulated physical form. The compounds of the present invention show a specific level of selectivity for PI3Kα, β, γ and δ of different paralogs. In particular, a specific level of selectivity for PI3Kγ is shown.

[0009] The compounds, compositions and methods described in the present invention directly correspond to the above needs and other purposes. Specifically, the present invention provides a class of compounds that can inhibit, regulate and / or modulate PI3-kinase activity, which are used to treat and / or prevent diseases, disorders, and / or conditions associated with abnormal PI3-kinase. Compared with existing similar compounds, the compounds of the present invention have better pharmacological activity. Specifically, the compounds of the present invention show excellent inhibitory activity against PI3-kinase, and are highly selective for PI3Kγ, have obvious advantages in liver microsomal stability, and have good pharmacokinetic properties and higher bioavailability. Therefore, the compounds of the present invention have very good development prospects. Summary of the invention

[0010] Definition of terms

[0011] Certain embodiments of the present invention are now described in detail, and examples thereof are illustrated by the accompanying structural formula and chemical formula. The present invention is intended to cover all substitutions, modifications and equivalent technical solutions, which are all included within the scope of the present invention. It should be appreciated by those skilled in the art that many methods and materials similar or equivalent to those described in the present invention can be used to practice the present invention. The present invention is by no means limited to the methods and materials described in the present invention. In the case where one or more of the combined documents, patents and similar materials are different from or contradictory to the present application (including but not limited to defined terms, term applications, described technologies, etc.), the present application shall prevail.

[0012] "Stereoisomers" refer to compounds that have the same chemical constitution but differ in the way the atoms or groups are arranged in space. Stereoisomers include enantiomers, diastereomers, conformational isomers (rotamers), geometric isomers (cis / trans) isomers, atropisomers, and the like.

[0013] As described herein, the compounds of the present invention may be optionally substituted with one or more substituents, such as the general compounds of the present invention, or as the specific examples, subclasses, and classes of compounds included in the embodiments.

[0014] It should be understood that the term "optionally substituted" can be used interchangeably with the term "substituted or unsubstituted". In general, the term "substituted" means that one or more hydrogen atoms in a given structure are replaced by a specific substituent, whether the hydrogen atom is attached to C or N or other atoms. "Optionally" unless otherwise indicated, an optional substituent group can be substituted at each substitutable position of the group. When more than one position in a given structural formula can be substituted by one or more substituents selected from a specific group, the substituents can be substituted at each position in the same or different manner.

[0015] The term "optionally substituted by..." can be used interchangeably with the term "unsubstituted or substituted by...", i.e., the structure is unsubstituted or substituted by one or more substituents described in the present invention, and the substituents described in the present invention include, but are not limited to, H, D, oxo (=O), F, Cl, Br, I, -OH, -CN, -NO2, -NR e R f 、-C(=O)R 7 、-OC(=O)R 7 、-C(=O)OR 7a 、-S(=O) 0-2 R 7 、-OS(=O) 1-2 R 7 、-S(=O) 1-2 OR 7a 、-N(R 8a )C(=O)R 8 、-C(=O)NR 8a R 8 、-OC(=O)NR 8a R 8 、-N(R 8a )S(=O) 1-2 R 8 、-S(=O) 1-2 NR 8a R 8 、-N(R 8a )C(=O)NR 8a R 8 , C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Haloalkyl, C1-6 Hydroxyalkyl, C 1-6 Aminoalkyl, C 1-6 Cyanoalkyl, C 1-6 Alkoxy, C 1-6 Alkylamino, C 3-8 Cycloalkyl, C 3-8 Cycloalkyl C 1-6 Alkyl, C 2-7 Heterocyclic group, C 2-7 Heterocyclyl C 1-6 Alkyl, C 6-12 Aryl, C 6-12 Aryl C 1-6 Alkyl, C 1-9 Heteroaryl, or C 1-9 Heteroaryl C 1-6 wherein each substituent is independently optionally replaced by 0, 1, 2, 3 or 4 independently selected from H, D, oxo (=O), F, Cl, Br, -OH, -NH2, -CN, -NO2, C 1-6 Deuterated alkyl, C 1-6 Alkyl and C 1-6 Alkoxy group substitution, etc. Among them, R e , R f , R 9 , R 9a , R 10 and R 10a Has the definition as described in the present invention.

[0016] In various parts of this specification, the substituents of the disclosed compounds of the present invention are disclosed according to group types or ranges. It is particularly noted that the present invention includes each independent secondary combination of the individual members of these group types and ranges. For example, the term "C1-C6 alkyl" specifically refers to the independently disclosed methyl, ethyl, C3 alkyl, C4 alkyl, C5 alkyl and C6 alkyl.

[0017] The term "alkyl" or "alkyl group" as used herein refers to a saturated straight or branched monovalent hydrocarbon group containing 1 to 20 carbon atoms, wherein the alkyl group may be optionally substituted with one or more substituents described herein. Unless otherwise specified, the alkyl group contains 1-20 carbon atoms. In one embodiment, the alkyl group contains 1-12 carbon atoms; in another embodiment, the alkyl group contains 1-6 carbon atoms; in yet another embodiment, the alkyl group contains 1-4 carbon atoms; in yet another embodiment, the alkyl group contains 1-3 carbon atoms. The alkyl group may be optionally substituted with one or more substituents described herein.

[0018] Examples of alkyl groups include, but are not limited to, methyl (Me, -CH3), ethyl (Et, -CH2CH3), n-propyl (n-Pr, -CH2CH2CH3), isopropyl (i-Pr, -CH(CH3)2), n-butyl (n-Bu, -CH2CH2CH2CH3), isobutyl (i-Bu, -CH2CH(CH3)2), sec-butyl (s-Bu, -CH(CH3)CH2CH3), tert-butyl (t-Bu, -CH(CH3)CH2CH3), u, -C(CH3)3), n-pentyl (-CH2CH2CH2CH2CH3), 2-pentyl (-CH(CH3)CH2CH2CH3), 3-pentyl (-CH(CH2CH3)2), 2-methyl-2-butyl (-C(CH3)2CH2CH3), 3-methyl-2-butyl (-CH(CH3)CH(CH3)2), 3-methyl-1-butyl (-CH2CH2CH(CH3)2), 2-methyl-1 -butyl (-CH2CH(CH3)CH2CH3), n-hexyl (-CH2CH2CH2CH2CH2CH3), 2-hexyl (-CH(CH3)CH2CH2CH2CH3), 3-hexyl (-CH(CH2CH3)(CH2CH2CH3)), 2-methyl-2-pentyl (-C(CH3)2CH2CH2CH3), 3-methyl-2-pentyl (-CH(CH3)CH(CH3)CH2CH3 ), 4-methyl-2-pentyl (-CH(CH3)CH2CH(CH3)2), 3-methyl-3-pentyl (-C(CH3)(CH2CH3)2), 2-methyl-3-pentyl (-CH(CH2CH3)CH(CH3)2), 2,3-dimethyl-2-butyl (-C(CH3)2CH(CH3)2), 3,3-dimethyl-2-butyl (-CH(CH3)C(CH3)3), n-heptyl, n-octyl, and the like.

[0019] The term "alkenyl" refers to a straight or branched chain monovalent hydrocarbon radical containing 2 to 12 carbon atoms and having at least one site of unsaturation, i.e., a carbon-carbon sp 2 Double bonds, including "cis" and "trans" orientations, or "E" and "Z" orientations. In one embodiment, the alkenyl group contains 2-8 carbon atoms; in another embodiment, the alkenyl group contains 2-6 carbon atoms; in yet another embodiment, the alkenyl group contains 2-4 carbon atoms. Examples of alkenyl groups include, but are not limited to, vinyl (-CH=CH2), allyl (-CH2CH=CH2), and the like. The alkenyl group may be optionally substituted with one or more substituents described herein.

[0020] The term "alkynyl" refers to a straight or branched monovalent hydrocarbon group containing 2-12 carbon atoms, wherein there is at least one unsaturated site, i.e., one carbon-carbon sp triple bond. In one embodiment, the alkynyl group contains 2-8 carbon atoms; in another embodiment, the alkynyl group contains 2-6 carbon atoms; in yet another embodiment, the alkynyl group contains 2-4 carbon atoms. Examples of alkynyl groups include, but are not limited to, ethynyl (-C≡CH), propargyl (-CH2C≡CH), 1-propynyl (-C≡C-CH3), and the like. The alkynyl group may be optionally substituted with one or more substituents described herein.

[0021] The term "alkoxy" means an alkyl group attached to the rest of the molecule via an oxygen atom, wherein the alkyl group has the definition as described herein. Unless otherwise specified, the alkoxy group contains 1-12 carbon atoms. In one embodiment, the alkoxy group contains 1-6 carbon atoms; in another embodiment, the alkoxy group contains 1-4 carbon atoms; in yet another embodiment, the alkoxy group contains 1-3 carbon atoms. The alkoxy group may be optionally substituted with one or more substituents described herein.

[0022] Examples of alkoxy groups include, but are not limited to, methoxy (MeO, -OCH3), ethoxy (EtO, -OCH2CH3), 1-propoxy (n-PrO, n-propoxy, -OCH2CH2CH3), 2-propoxy (i-PrO, i-propoxy, -OCH(CH3)2), 1-butoxy (n-BuO, n-butoxy, -OCH2CH2CH2CH3), 2-methyl-1-propoxy (i-BuO, i-butoxy, -OCH2CH(CH3)2), 2-butoxy (s-BuO, s-butoxy, -OCH(CH3)CH2CH3), 2-methyl-2- Propoxy (t-BuO, t-butoxy, -OC(CH3)3), 1-pentyloxy (n-pentyloxy, -OCH2CH2CH2CH2CH3), 2-pentyloxy (-OCH(CH3)CH2CH2CH3), 3-pentyloxy (-OCH(CH2CH3)2), 2-methyl-2-butoxy (-OC(CH3)2CH2CH3), 3-methyl-2-butoxy (-OCH(CH3)CH(CH3)2), 3-methyl-l-butoxy (-OCH2CH2CH(CH3)2), 2-methyl-l-butoxy (-OCH2CH(CH3)CH2CH3), and the like.

[0023] The term "haloalkyl" or "haloalkoxy" means that an alkyl or alkoxy group is substituted by one or more halogen atoms. Examples of such groups include, but are not limited to, trifluoromethyl (-CF3), trifluoromethoxy (-OCF3), difluoroethyl (-CH2CHF2, -CF2CH3, -CHFCH2F), trifluoroethyl (-CH2CF3, -CF2CH2F, -CFHCHF2), -CF(CH3)2, etc.

[0024] The terms "deuterated alkyl", "deuterated alkoxy", "deuterated cycloalkyl", "deuterated heterocyclyl", "deuterated aryl", or "deuterated heteroaryl" mean that an alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl group is substituted with one or more D atoms. Examples of such groups include, but are not limited to, monodeuterated methyl (-CH2D), dideuterated methyl (-CHD2), trideuterated methyl (-CD3), monodeuterated methoxy (-OCH2D), dideuterated methoxy (-OCHD2), trideuterated methoxy (-OCD3), dideuterated ethyl (-CH2CHD2, -CD2CH3, -CHDCH2D), pentadeuterated ethyl (-CD2CD3), deuterated cyclopropyl, deuterated cyclohexyl, pentadeuterated phenyl, and the like.

[0025] The term "deuterium (D)-containing group" refers to a group or a portion of a group described in the present invention in which one or more H atoms are replaced by D, but does not include a single D group. Such groups include, but are not limited to, C 1-6 Deuterated alkyl, C 3-8 Deuterated cycloalkyl, C 2-9 Deuterated heterocyclic group, C 6-10 Deuterated aryl, C 1-9 deuterated heteroaryl, etc., each of which has the definition described in the present invention, wherein each group is independently and optionally replaced by one or more H, D, F, Cl, Br, I, -OH, -CN, -NO2, -NR e R f , C 1-6 Alkyl and C 1-6 is substituted with a deuterated alkyl group, wherein R e and R f All have the definitions described in the present invention.

[0026] The terms "hydroxyalkyl" or "hydroxy-substituted alkyl" and "hydroxyalkoxy" or "hydroxy-substituted alkoxy" refer to an alkyl or alkoxy group, respectively, as the case may be, substituted with one or more hydroxy groups, wherein "hydroxyalkyl" and "hydroxyalkyl" are used interchangeably, such examples include, but are not limited to, hydroxymethyl (-CH2OH), 2-hydroxyethyl (-CH2CH2OH), 1-hydroxyethyl (-CH(OH)CH3), 2-hydroxyprop-2-yl (-COH(CH3)2), 2-hydroxy-2-methylpropyl (-CH2COH(CH3)2), 3-hydroxypropyl (-CH2CH2CH2OH), 2-hydroxypropyl (-CH2CH(OH)CH3), 2-hydroxy-2-methylpropyl (-CH2CH(OH)(CH3)CH3), hydroxymethoxy (-OCH2OH), and the like.

[0027] The term "cyano-substituted alkyl" or "cyanoalkyl" includes C substituted with one or more cyano groups. 1-10 In some embodiments, the cyanoalkyl group is a C substituted with one or more cyano groups. 1-6 "Lower cyanoalkyl" is a C substituted with one or more cyano groups. 1-4 Examples of “lower cyanoalkyl” include, but are not limited to, CNCH2-, CNCH2CH2-, CNCH2CH2CH2-, CNCH2CHCNCH2-, and the like.

[0028] The term "alkylamino" includes "N-alkylamino" and "N,N-dialkylamino", wherein the amino groups are independently substituted with one or two alkyl groups. In some embodiments, the alkylamino group is one or two C 1-6 The alkyl group is a lower alkylamino group attached to the nitrogen atom. In some other embodiments, the alkylamino group is C 1-3 Suitable alkylamino groups may be monoalkylamino or dialkylamino, examples of which include, but are not limited to, N-methylamino, N-ethylamino, N,N-dimethylamino, N,N-diethylamino, and the like.

[0029] The term "aminoalkyl" includes C 1-10 In some embodiments, aminoalkyl is a C substituted with one or more amino groups. 1-6 "Lower aminoalkyl" is another embodiment of aminoalkyl being a C substituted with one or more amino groups. 1-4 Examples of "lower aminoalkyl" include, but are not limited to, aminomethyl, aminoethyl, aminopropyl, aminobutyl and aminohexyl.

[0030] The term "cycloalkyl" refers to a monovalent or multivalent saturated monocyclic, bicyclic or tricyclic ring system containing 3-12 carbon atoms. Bicyclic cycloalkyls include spirobicyclic alkyls, fused bicyclic alkyls and bridged bicyclic alkyls. In some embodiments, the cycloalkyl contains 3-12 carbon atoms; in other embodiments, the cycloalkyl contains 3-10 carbon atoms; in other embodiments, the cycloalkyl contains 3-8 carbon atoms; in other embodiments, the cycloalkyl contains 3-7 carbon atoms; in other embodiments, the cycloalkyl contains 3-6 carbon atoms; in still other embodiments, the cycloalkyl is C7-C 12 Cycloalkyl containing C7-C 12 Monocyclic alkyl, C7-C 12 Bicyclic alkyl (such as C7-C 12 Spirobicycloalkyl, C7-C 12 Fused bicycloalkyl and C7-C 12 bridged bicycloalkyl) or C7-C 12 Tricycloalkyl. The cycloalkyl group can be independently unsubstituted or substituted by one or more substituents described in the present invention. The term "monocyclic cycloalkyl" or "monocyclic alkyl" represents a cycloalkyl of a monocyclic system, wherein the cycloalkyl has the definition as described above, and the monocyclic cycloalkyl group can be independently unsubstituted or substituted by one or more substituents described in the present invention. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopentyl-1-alkenyl, 1-cyclopentyl-2-alkenyl, 1-cyclopentyl-3-alkenyl, cyclohexyl, 1-cyclohexyl-1-alkenyl, 1-cyclohexyl-2-alkenyl, 1-cyclohexyl-3-alkenyl, cyclohexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, cyclododecyl, etc.

[0031] The term "cycloalkylalkyl" includes cycloalkyl-substituted alkyl groups. In some embodiments, the cycloalkylalkyl group refers to a "lower cycloalkylalkyl" group, i.e., a cycloalkyl group attached to a C 1-6 In other embodiments, a cycloalkylalkyl group refers to a C-containing 1-3 Specific examples include, but are not limited to, cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl, cyclohexylmethyl, cyclopentylethyl, cyclohexylethyl, etc. The cycloalkyl on the cycloalkylalkyl group may be further substituted with one or more substituents described herein.

[0032] The terms "heterocyclyl" and "heterocycle" are used interchangeably herein and refer to a monocyclic, bicyclic or tricyclic ring system containing 3-12 ring atoms, monovalent or polyvalent, saturated or partially unsaturated, non-aromatic, wherein at least one ring atom is selected from nitrogen, oxygen and sulfur atoms. In some embodiments, the heterocyclyl or heterocycle contains 4-12 ring atoms. In some embodiments, the heterocyclyl or heterocycle contains 5-12 ring atoms. In some embodiments, the heterocyclyl or heterocycle contains 4-8 ring atoms. In some embodiments, the heterocyclyl or heterocycle contains 3-10 ring atoms. In some embodiments, the heterocyclyl or heterocycle contains 3-8 ring atoms. In some embodiments, the heterocyclyl or heterocycle contains 3-6 ring atoms. In some embodiments, the heterocyclyl or heterocycle contains 4-7 ring atoms. Unless otherwise specified, the heterocyclic group can be a carbon group or a nitrogen group, and the -CH2- group can be optionally replaced by -C(=O)-, the sulfur atom of the ring can be optionally oxidized to S-oxide, and the nitrogen atom of the ring can be optionally oxidized to N-oxygen compound. The heterocyclic group includes saturated heterocyclic groups (heterocycloalkyl) and partially unsaturated heterocyclic groups. The heterocyclic group has one or more points of attachment to the rest of the molecule. Examples of heterocyclic groups include, but are not limited to, oxirane, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrrolinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, dihydrothiophenyl, 1,3-dioxolane, dithiolanyl, tetrahydropyranyl, dihydropyranyl, 2H-pyranyl, 4H-pyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, dioxanyl, dithianyl, thioxanyl, homopiperazinyl, homopiperidinyl, oxepanyl, thiepanyl, azepanyl, oxazepanyl, oxe ... Base (eg, 1,4-oxazepine 1,2-oxazepine , diazepine Base (eg, 1,4-diazepine Base, 1,2-diazepine , dioxa Base (such as 1,4-dioxa 1,2-dioxa , thiazolin Base (such as 1,4-thiazepine 1,2-thiazepine 1,2,3,4-tetrahydroisoquinolinyl, 1,3-benzodioxolyl, 2-oxa-5-azabicyclo[2.2.1]hept-5-yl, 2-azaspiro[4.4]nonyl, 1,6-dioxaspiro[4.4]nonyl, 2-azaspiro[4.5]decyl, 8-azaspiro[4.5]decyl, 7-azaspiro[4.5]decyl, 3-azaspiro[5.5]undecyl, 2-azaspiro[5.5]undecyl, octahydro-1H-isoindolyl, octahydrocyclopenta[c]pyrrolyl, dihydroindolyl, 1,2,3,4-tetrahydroisoquinolinyl, hexahydrofuro[3,2-b]furanyl and dodecahydroisoquinolinyl, and the like. Examples of heterocyclic groups in which the -CH2- group is replaced by -C(=O)- include, but are not limited to, 2-oxopyrrolidinyl, oxo-1,3-thiazolidinyl, 2-piperidone and 3,5-dioxopiperidinyl. Examples of heterocyclic groups in which the sulfur atom is oxidized include, but are not limited to, cyclopentane sulfone and 1,1-dioxothiomorpholinyl. The heterocyclic group may be optionally substituted by one or more substituents described herein.

[0033] In one embodiment, the heterocyclic group is a 4-7 atom heterocyclic group, which refers to a monovalent or polyvalent, saturated or partially unsaturated non-aromatic monocyclic or bicyclic ring containing 4-7 ring atoms, wherein at least one ring atom is selected from nitrogen, sulfur and oxygen atoms. Unless otherwise specified, the 4-7 atom heterocyclic group can be a carbon group or a nitrogen group, and the -CH2- group can be optionally replaced by -C(=O)-. The sulfur atom of the ring can be optionally oxidized to an S-oxide. The nitrogen atom of the ring can be optionally oxidized to an N-oxygen compound. The 4-7 atom heterocyclic group has one or more points of attachment to the rest of the molecule. Examples of monocyclic heterocyclic groups of 4 to 7 atoms include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrrolinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, dihydrothiophenyl, tetrahydropyranyl, dihydropyranyl, 2H-pyranyl, 4H-pyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, dioxanyl, dithianyl, thioxanyl, 1,2-oxazinyl, 1,2-thiazinyl, hexahydropyridazinyl, homopiperazinyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepanyl, 1,4-oxazepine 1,2-oxazepine , diazepine 1,4-diazepine Base, 1,2-diazepine yl) and thiazepine 1,4-thiazepine 1,2-thiazepine Examples of bicyclic heterocyclic groups composed of 4 to 7 atoms include, but are not limited to: 3-azabicyclo[3,2,0]heptane, 3-oxobicyclo[3,2,0]heptane, etc.; Examples of heterocyclic groups composed of 4 to 7 atoms in which -CH2- groups are replaced by -C(=O)- include, but are not limited to, 2-oxopyrrolidinyl, oxo-1,3-thiazolidinyl, 2-piperidone and 3,5-dioxopiperidinyl; Examples of heterocyclic groups composed of 4 to 7 atoms in which sulfur atoms are oxidized include, but are not limited to, cyclopentanesulfonyl, 1,1-dioxotetrahydrothiophene, 1,1-dioxotetrahydrothiopyran, 1,1-dioxothiomorpholinyl. The heterocyclic groups composed of 4 to 7 atoms may be optionally substituted by one or more substituents described in the present invention.

[0034] The term "heterocyclylalkyl" includes alkyl groups substituted with heterocyclyl, wherein heterocyclyl and alkyl have the meanings described herein, such examples include, but are not limited to, tetrahydrofuranylmethyl, pyrrol-2-ylmethyl, morpholin-4-ylethyl, piperazin-4-ylethyl, piperidin-4-ylethyl, and the like.

[0035] The term "aryl" refers to monocyclic, bicyclic and tricyclic carbon ring systems containing 6-14 ring atoms, or 6-12 ring atoms, or 6-10 ring atoms, wherein at least one ring system is aromatic, wherein each ring system contains a ring consisting of 3-7 atoms and has one or more points of attachment to the rest of the molecule. The term "aryl" can be used interchangeably with the term "aromatic ring". Examples of aryl groups can include phenyl, naphthyl and anthracenyl. The aryl groups can be independently optionally substituted with one or more substituents described herein.

[0036] The term "arylalkyl" or "aralkyl" includes alkyl groups substituted with aryl. In some embodiments, the arylalkyl group refers to a "lower arylalkyl" group, i.e., an aryl group is attached to a C 1-6 In other embodiments, the arylalkyl group refers to a C 1-3 Specific examples include, but are not limited to, benzyl, diphenylmethyl, phenethyl, etc. The aryl group on the arylalkyl group may be further substituted with one or more substituents described in the present invention.

[0037] The term "heteroaryl" refers to monocyclic, bicyclic and tricyclic ring systems containing 5-12 ring atoms, or 5-10 ring atoms, or 5-6 ring atoms, wherein all rings are aromatic and at least one aromatic ring contains one or more heteroatoms, wherein each ring system contains 5-7 ring atoms and has one or more points of attachment to the rest of the molecule. The term "heteroaryl" can be used interchangeably with the terms "heteroaromatic ring" or "heteroaromatic compound". In one embodiment, heteroaryl is a heteroaryl containing 5-12 atoms of 1, 2, 3 or 4 heteroatoms independently selected from O, S and N. In another embodiment, heteroaryl is a heteroaryl containing 5-10 atoms of 1, 2, 3 or 4 heteroatoms independently selected from O, S and N. In another embodiment, heteroaryl is a heteroaryl containing 5-6 atoms of 1, 2, 3 or 4 heteroatoms independently selected from O, S and N. The heteroaryl group is optionally substituted with one or more substituents described herein.

[0038] Examples of heteroaryl groups include, but are not limited to, 2-furanyl, 3-furanyl, N-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, N-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 1,2,3-oxadiazolyl, 1,2,5-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,3-triazolyl , 1,2,3-thiodiazolyl, 1,3,4-thiodiazolyl, 1,2,5-thiodiazolyl, pyrazinyl, 1,3,5-triazinyl; also includes the following bicyclic rings, but is by no means limited to these bicyclic rings: benzimidazolyl, benzofuranyl, benzothiophenyl, indolyl (such as 2-indolyl), purinyl, quinolyl (such as 2-quinolyl, 3-quinolyl, 4-quinolyl), isoquinolyl (such as 1-isoquinolyl), [1,2,4]triazolo[4,3-b]pyridazinyl, [1,2,4]triazolo[1,5-a]pyrimidinyl, [1,2,4]triazolo[1,5-a]pyridinyl, and the like.

[0039] The term "heteroarylalkyl" means an alkyl group substituted by one or more heteroaryl groups, wherein the heteroaryl and alkyl groups have the meanings described herein, examples of which include, but are not limited to, pyridine-2-methyl, imidazole-2-methyl, furan-2-ethyl, indole-3-methyl, and the like.

[0040] The term "halogen" refers to F, Cl, Br or I.

[0041] As described in the present invention, there are two connection points in the ring system connected to the rest of the molecule, as shown in formula (a1), indicating that either the E end or the E' end can be connected to the rest of the molecule, that is, the connection methods of the two ends can be interchangeable.

[0042] E-(CH2) t1 -LE′ (a1)

[0043] As described in the present invention, the substituent is connected to the central ring by a bond to form a ring system (as shown below) to represent that the substituent can be substituted at any substitutable position on any ring. For example, formula b represents that any possible substitutable position on ring A or ring B can be substituted, as shown in formula c, d, e, f, g, h, i, j, k, l, m, n, o, p, q, etc.

[0044]

[0045] The "pharmaceutically acceptable salt" used in the present invention refers to organic salts and inorganic salts of the compounds of the present invention. Pharmaceutically acceptable salts are well known in the art, as described in the literature: SM Berge et al., describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66: 1-19. Pharmaceutically acceptable salts formed by non-toxic acids include, but are not limited to, inorganic acid salts formed by reaction with amino groups, such as hydrochlorides, hydrobromides, phosphates, sulfates, perchlorates, and organic acid salts such as acetates, oxalates, maleates, tartrates, citrates, succinates, malonates, or other methods described in books and literature, such as ion exchange methods, to obtain these salts. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, cyclopentylpropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, picrate, pivalate, propionate, stearate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like. Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N + (C 1-4 The present invention also contemplates quaternary ammonium salts formed by any compound containing a N group. Water-soluble or oil-soluble or dispersible products can be obtained by quaternization. Alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Pharmaceutically acceptable salts further include appropriate, non-toxic ammonium, quaternary ammonium salts and amine cations formed by counter ions, such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, C 1-8 Sulfonates and aromatic sulfonates.

[0046] The "solvate" of the present invention refers to an association formed by one or more solvent molecules and the compound of the present invention. Solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid and aminoethanol. The term "hydrate" refers to an association formed by the solvent molecule being water.

[0047] Description of the compounds of the present invention

[0048] The present invention discloses a novel class of pyrimidine amine compounds, which can be used as inhibitors of PI3-kinase activity, especially PI3K-γ activity, and are used to prevent, treat, cure, and / or alleviate diseases, disorders, and / or conditions associated with abnormal PI3-kinase, such as respiratory diseases, viral infections, non-viral respiratory infections, allergic diseases, autoimmune diseases, inflammatory diseases, cardiovascular diseases, malignant blood diseases, neurodegenerative diseases, pancreatitis, multiple organ failure, kidney disease, platelet aggregation, cancer, sperm motility, transplant rejection, graft rejection, lung injury or pain, etc. Compared with existing similar compounds, the compounds of the present invention have better pharmacological activity, specifically, the compounds of the present invention show excellent inhibitory activity against PI3-kinase and are highly selective against PI3Kγ, and show obvious advantages in pharmacokinetics. Therefore, the compounds of the present invention have very good development prospects.

[0049] The compounds disclosed in the present invention can show strong inhibitory activity against PI3-kinase, especially PI3K-γ. On the one hand, the present invention provides an isoquinolinone and quinazolinone compound having a structure shown in formula (I):

[0050]

[0051] or a stereoisomer, tautomer, deuterated form, nitrogen oxide, solvate, or pharmaceutically acceptable salt thereof;

[0052] in,

[0053] X is -C(R c )-, or N;

[0054] Z 1 and Z 2 Each is independently -C(R 4d )-or N;

[0055] R a , R b and R c Each independently represents H, D, F, Cl, C 1-3 Alkyl, C 1-6 Deuterated alkyl, C 1-3 Alkoxy, C 1-3 Haloalkyl, C 1-3 Hydroxyalkyl, C 1-3 Aminoalkyl, C 1-3 Cyanoalkyl, C 1-6 Deuterated alkoxy, C substituted with 1 to 5 deuterium 1-6 Haloalkyl, C substituted with 1 to 5 deuterium 1-6 Hydroxyalkyl, C substituted with 1 to 5 deuterium 3-8Cycloalkyl, or C 1-9 heteroaryl;

[0056] R 1 It is H, D, -C 1-6 Alkyl-NR e R f 、-C(=O)R 7 、-C(=O)OR 7a 、-C(=O)NR 7 R 7a , C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Haloalkyl, C 3-8 Cycloalkyl, C 2-9 Heterocyclic group, C 6-10 Aryl, or C 1-9 Heteroaryl; wherein R 1 Optionally, 0, 1, 2, 3 or 4 R 5 replace;

[0057] R 2 Each occurrence is independently H, D, F, Cl, Br, I, -OH, -CN, -NO2, -NR e R f , C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy, C 1-6 Hydroxyalkyl, C 1-6 Aminoalkyl, C 1-6 Cyanoalkyl, C 2-9 Heterocyclic group, C 3-8 Cycloalkyl, C 6-10 Aryl, or C 1-9 Heteroaryl; wherein R 2 At each occurrence, 0, 1, 2, 3, 4 or 5 independently selected from H, D, oxo (=O), F, Cl, Br, I, -OH, -NH2, -CN, -NO2, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Alkoxy and C 1-6 haloalkyl radical substitution;

[0058] R 3 It is H, D, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6Haloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-8 Cycloalkyl, C 3-8 Deuterated cycloalkyl, C 2-9 Heterocyclic group, C 6-10 Aryl, or C 1-9 Heteroaryl; wherein R 3 Optionally, 0, 1, 2, 3, 4 or 5 R 6 replace;

[0059] R 4a , R 4b , R 4c and R 4d are independently H, D, F, Cl, -CN, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy, C 1-6 Hydroxyalkyl, C 1-6 Aminoalkyl, C 1-6 Cyanoalkyl, C 2-9 Heterocyclic group, C 3-8 Cycloalkyl, C 6-10 Aryl, or C 1-9 Heteroaryl; wherein R 4a , R 4b , R 4c and R 4d Each is independently optionally replaced by 0, 1, 2, 3, 4 or 5 independently selected from H, D, oxo (=O), F, Cl, Br, I, -OH, -NH2, -CN, -NO2, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Alkoxy, C 1-6 Deuterated alkoxy, C 1-6 Haloalkyl and C 1-6 radical substitution with halodeuterated alkyl;

[0060] R 5 and R 6 , each occurrence is independently H, D, oxo (=O), F, Cl, Br, I, -OH, -CN, -NO2, -NR e R f 、-C(=O)R 7 , C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy, C 1-6 Hydroxyalkyl, C 1-6 Aminoalkyl, C 1-6 Cyanoalkyl, C 2-9 Heterocyclic group, C 3-8 Cycloalkyl, C 6-10 Aryl, or C 1-9 heteroaryl; wherein each -NR e R f 、-C(=O)R 7 , C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy, C 1-6 Hydroxyalkyl, C 1-6 Aminoalkyl, C 1-6 Cyanoalkyl, C 2-9 Heterocyclic group, C 3-8 Cycloalkyl, C 6-10 Aryl and C 1-9 Heteroaryl is optionally substituted by 0, 1, 2, 3, 4 or 5 groups independently selected from H, D, oxo (=O), F, Cl, Br, I, -OH, -NH2, -CN, -NO2, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Alkoxy and C 1-6 haloalkyl radical substitution;

[0061] R e , R f , R 7 and R 7a , at each occurrence, are independently H, D, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl, C 3-8 Cycloalkyl, C 3-8 Cycloalkyl C 1-6 Alkyl, C 2-9 Heterocyclic group, C 2-9 Heterocyclyl C 1-6 Alkyl, C 6-10 Aryl, C 6-10 Aryl C 1-6 Alkyl, C 1-9 Heteroaryl, or C 1-9 Heteroaryl C 1-6Alkyl; wherein each C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl, C 3-8 Cycloalkyl, C 3-8 Cycloalkyl C 1-6 Alkyl, C 2-9 Heterocyclic group, C 2-9 Heterocyclyl C 1-6 Alkyl, C 6-12 Aryl, C 6-12 Aryl C 1-6 Alkyl, C 1-9 Heteroaryl and C 1-9 Heteroaryl C 1-6 The alkyl radical is optionally substituted with 0, 1, 2, 3, 4 or 5 radicals independently selected from H, D, oxo (=O), F, Cl, Br, I, -OH, -NH2, -CN, -NO2, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Alkoxy and C 1-6 haloalkyl radical substitution; and

[0062] n is 0, 1, 2, or 3;

[0063] The conditions are: 1) When R 1 When it is pyrazolyl, R 1 Optionally, 0, 1, 2, 3 or 4 R 5 Replace, then each R 2 , R 4a , R 4b and R 4c At least one of them is independently F, Cl, -CN, C 1-6 Alkoxy, C 1-6 Deuterated alkyl, C 1-6 Deuterated alkoxy, C substituted with 1 to 5 deuterium 1-6 Haloalkyl, C substituted with 1 to 5 deuterium 1-6 Hydroxyalkyl, C substituted with 1 to 5 deuterium 3-8 Cycloalkyl, or C 1-9 heteroaryl; or 2) when R 1 It is H, D, -C 1-6 Alkyl-NR e R f 、-C(=O)R 7 、-C(=O)OR 7a 、-C(=O)NR 7 R 7a , C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Haloalkyl, C3-8 Cycloalkyl, C 2-9 Heterocyclic group, C 6-10 Aryl, or C except pyrazolyl 1-9 When heteroaryl, R 1 Optionally, 0, 1, 2, 3 or 4 R 5 Replace, then each R a , R b , R c , R 2 , R 3 , R 4a , R 4b , R 4c and R 4d At least one of them is independently D (deuterium), C 1-6 Deuterated alkyl, C 1-6 Deuterated alkoxy, C substituted with 1 to 5 deuterium 1-6 Haloalkyl, C substituted with 1 to 5 deuterium 1-6 Hydroxyalkyl, C substituted with 1 to 5 deuterium 3-8 Cycloalkyl, or C 1-9 Heteroaryl.

[0064] On the other hand, the present invention provides an isoquinolinone and quinazolinone compound having a structure shown in formula (Ib):

[0065]

[0066] or a stereoisomer, tautomer, deuterated form, nitrogen oxide, solvate, or pharmaceutically acceptable salt thereof;

[0067] in,

[0068] X is -C(R c )-, or N;

[0069] Z 1 and Z 2 Each is independently -C(R 4d )-or N;

[0070] R a , R b and R c Each independently represents H, D, F, Cl, C 1-3 Alkyl, C 1-3 Deuterated alkyl, C 1-3 Alkoxy, C 1-3 Haloalkyl, C 1-3 Hydroxyalkyl, C 1-3 Aminoalkyl, or C 1-3 Cyanoalkyl;

[0071] R1 It is H, D, -C 1-6 Alkyl-NR e R f 、-C(=O)R 7 、-C(=O)OR 7a 、-C(=O)NR 7 R 7a , C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Haloalkyl, C 3-8 Cycloalkyl, C 2-9 Heterocyclic group, C 6-10 Aryl, or C 1-9 Heteroaryl; wherein R 1 Optionally, 0, 1, 2, 3 or 4 R 5 replace;

[0072] R 2 Each occurrence is independently H, D, F, Cl, Br, I, -OH, -CN, -NO2, -NR e R f , C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy, C 1-6 Hydroxyalkyl, C 1-6 Aminoalkyl, C 1-6 Cyanoalkyl, C 2-9 Heterocyclic group, C 3-8 Cycloalkyl, C 6-10 Aryl, or C 1-9 heteroaryl; wherein each R 2 , independently and optionally replaced by 0, 1, 2, 3, 4 or 5 independently selected from H, D, oxo (=O), F, Cl, Br, I, -OH, -NH2, -CN, -NO2, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Alkoxy and C 1-6 haloalkyl radical substitution;

[0073] R 3 It is H, D, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Haloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-8 Cycloalkyl, C3-8 Deuterated cycloalkyl, C 2-9 Heterocyclic group, C 6-10 Aryl, or C 1-9 Heteroaryl; wherein R 3 Optionally, 0, 1, 2, 3, 4 or 5 R 6 replace;

[0074] R 4a , R 4b , R 4c and R 4d are each independently H, D, F, Cl, a deuterium-containing group, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy, C 1-6 Hydroxyalkyl, C 1-6 Aminoalkyl, C 1-6 Cyanoalkyl, C 2-9 Heterocyclic group, C 3-8 Cycloalkyl, C 6-10 Aryl, or C 1-9 Heteroaryl; wherein R 4a , R 4b , R 4c and R 4d Each is independently optionally replaced by 0, 1, 2, 3, 4 or 5 independently selected from H, D, oxo (=O), F, Cl, Br, I, -OH, -NH2, -CN, -NO2, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Alkoxy, C 1-6 Deuterated alkoxy, C 1-6 Haloalkyl and C 1-6 radical substitution with halodeuterated alkyl;

[0075] R 5 and R 6 , each occurrence is independently H, D, oxo (=O), F, Cl, Br, I, -OH, -CN, -NO2, -NR e R f 、-C(=O)R 7 , C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy, C 1-6Hydroxyalkyl, C 1-6 Aminoalkyl, C 1-6 Cyanoalkyl, C 2-9 Heterocyclic group, C 3-8 Cycloalkyl, C 6-10 Aryl, or C 1-9 heteroaryl; wherein each -NR e R f 、-C(=O)R 7 , C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy, C 1-6 Hydroxyalkyl, C 1-6 Aminoalkyl, C 1-6 Cyanoalkyl, C 2-9 Heterocyclic group, C 3-8 Cycloalkyl, C 6-10 Aryl and C 1-9 Heteroaryl is optionally substituted by 0, 1, 2, 3, 4 or 5 groups independently selected from H, D, oxo (=O), F, Cl, Br, I, -OH, -NH2, -CN, -NO2, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Alkoxy and C 1-6 haloalkyl radical substitution;

[0076] R e , R f , R 7 and R 7a , at each occurrence, are independently H, D, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl, C 3-8 Cycloalkyl, C 3-8 Cycloalkyl C 1-6 Alkyl, C 2-9 Heterocyclic group, C 2-9 Heterocyclyl C 1-6 Alkyl, C 6-10 Aryl, C 6-10 Aryl C 1-6 Alkyl, C 1-9 Heteroaryl, or C 1-9 Heteroaryl C 1-6 Alkyl; wherein each C 1-6 Alkyl, C 1-6 Haloalkyl, C1-6 Hydroxyalkyl, C 3-8 Cycloalkyl, C 3-8 Cycloalkyl C 1-6 Alkyl, C 2-9 Heterocyclic group, C 2-9 Heterocyclyl C 1-6 Alkyl, C 6-12 Aryl, C 6-12 Aryl C 1-6 Alkyl, C 1-9 Heteroaryl and C 1-9 Heteroaryl C 1-6 The alkyl radical is optionally substituted with 0, 1, 2, 3, 4 or 5 radicals independently selected from H, D, oxo (=O), F, Cl, Br, I, -OH, -NH2, -CN, -NO2, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Alkoxy and C 1-6 haloalkyl radical substitution; and

[0077] n is 0, 1, 2, or 3;

[0078] The conditions are: 1) When R 1 When it is pyrazolyl, R 2 , R 4a , R 4b and R 4c At least one of is independently F, Cl, or a deuterated group; or 2) when R 1 It is H, D, -C 1-6 Alkyl-NR e R f 、-C(=O)R 7 、-C(=O)OR 7a 、-C(=O)NR 7 R 7a , C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Haloalkyl, C 3-8 Cycloalkyl, C 2-9 Heterocyclic group, C 6-10 Aryl, or C except pyrazolyl 1-9 When heteroaryl, R a , R b , R c , R 2 , R 3 , R 4a , R 4b , R 4c and R 4d At least one of them is independently D (deuterium) or a deuterium-containing group.

[0079] In some embodiments, the compound represented by formula (I) has the structure represented by formula (Ia):

[0080]

[0081] or a stereoisomer, tautomer, deuterated substance, nitrogen oxide, solvate, or pharmaceutically acceptable salt thereof.

[0082] In some embodiments of Formula (I) or (Ia), R 1 is phenyl, pyridyl, pyridazinyl, pyrazinyl, pyrimidinyl, triazinyl, thiazolyl, imidazolyl, oxazolyl, thiadiazolyl,

[0083] where Y 1 is O, S, or -NH-;

[0084] Y 6 is O, or -NH-;

[0085] t3 and t4 are each independently 1, 2, 3 or 4; and

[0086] Where R 1 Optionally, 0, 1, 2, 3 or 4 R 5 replace;

[0087] The conditions are: 1) R 1 for When R 1 Optionally, 0, 1, 2, 3 or 4 R 5 Replace, then each R 2 , R 4a , R 4b and R 4c At least one of them is independently F, Cl, -CN, C 1-3 Alkoxy, C 1-3 Deuterated alkyl, C 1-3 Deuterated alkoxy, C substituted with 1 to 5 deuterium 1-3 Haloalkyl, C substituted with 1 to 5 deuterium 1-3 Hydroxyalkyl, C substituted with 1 to 5 deuterium 3-6 Cycloalkyl, or C 1-9 Heteroaryl; 2) when R 1 is phenyl, pyridyl, pyridazinyl, pyrazinyl, pyrimidinyl, triazinyl, thiazolyl, imidazolyl, oxazolyl, thiadiazolyl, When R 1 Optionally 0, 1, 2, 3 or 4 R 5 Replace, then each R a , R b , R c, R 2 , R 3 , R 4a , R 4b , R 4c and R 4d At least one of them is independently D (deuterium), C 1-3 Deuterated alkyl, C 1-3 Deuterated alkoxy, C substituted with 1 to 5 deuterium 1-3 Haloalkyl, C substituted with 1 to 5 deuterium 1-3 Hydroxyalkyl, C substituted with 1 to 5 deuterium 3-6 Cycloalkyl, or C 1-9 Heteroaryl.

[0088] In some embodiments of Formula (I) or (Ib), R 1 is phenyl, pyridyl, pyridazinyl, pyrazinyl, pyrimidinyl, triazinyl, thiazolyl, imidazolyl, oxazolyl, thiadiazolyl,

[0089] where Y 1 is O, S, or -NH-;

[0090] Y 6 is O, or -NH-;

[0091] t3 and t4 are each independently 1, 2, 3 or 4; and

[0092] Where R 1 Optionally 0, 1, 2, 3 or 4 R 5 replace;

[0093] The conditions are: 1) R 1 for When R 2 , R 4a , R 4b and R 4c At least one of them is independently F, Cl, or a deuterated group; 2) when R 1 is phenyl, pyridyl, pyridazinyl, pyrazinyl, pyrimidinyl, triazinyl, thiazolyl, imidazolyl, oxazolyl, thiadiazolyl, When R a , R b , R c , R 2 , R 3 , R 4a , R 4b , R 4c and R 4d At least one of them is independently D (deuterium) or a deuterium-containing group.

[0094] In some embodiments of Formula (I) or (Ia), R 4a , R 4b , R 4c and R 4d Each independently represents H, D, F, Cl, C 1-3 Alkyl, C 1-3 Deuterated alkyl, C 1-3 Alkoxy, C 1-3 Haloalkyl, C 1-3 Halogenated alkoxy, C 1-3 Hydroxyalkyl, C 1-3 Aminoalkyl, C 1-3 Cyanoalkyl, C 3-6 Heterocyclic group, C 3-6 Deuterated heterocyclic group, C 3-6 Cycloalkyl, C 3-6 deuterated cycloalkyl, phenyl, or a heteroaryl group consisting of 5-6 atoms; wherein R 4a , R 4b , R 4c and R 4d Each is independently optionally replaced by 0, 1, 2, 3, 4 or 5 independently selected from H, D, oxo (=O), F, Cl, Br, I, -OH, -NH2, -CN, -NO2, C 1-3 Alkyl, C 1-3 Deuterated alkyl, C 1-3 Alkoxy, C 1-3 Deuterated alkoxy, C 1-3 Haloalkyl and C 1-3 The radicals are substituted with halo-deuterated alkyl.

[0095] In some embodiments of Formula (I), (Ia) or (Ib), R 4a , R 4b , R 4c and R 4d Each independently is H, F, Cl, D, -CN, -OCH3, -OCH2CH3,

[0096] On the other hand, the present invention also provides a compound represented by formula (II):

[0097]

[0098] or a stereoisomer, tautomer, deuterated form, nitrogen oxide, solvate, or pharmaceutically acceptable salt thereof;

[0099] in,

[0100] X is -C(R c )-, or N;

[0101] W is for C 3-10 Cycloalkyl, C 2-9 Heterocyclic group, C 6-10 Aryl, or C 1-9 Heteroaryl; wherein W is optionally replaced by 0, 1, 2, 3 or 4 R 4 replace;

[0102] R a , R b and R c Each independently represents H, D, F, Cl, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl, C 1-6 Aminoalkyl, or C 1-6 Cyanoalkyl;

[0103] R 1 Yes-C 1-3 Hydroxyalkyl-C(=O)OR 7a , -C(=O)OR 7a , -C(=O)NR 7 R 7a ,

[0104] where Y 2 and Y 3 Each independently is -(CH2) t1 -、-(CH2) t1 -L-, -(CH2) t1 -L-(CH2) t2 -, O, or -NH-;

[0105] Y 4 Yes - (CH2) t1 -、-(CH2) t1 -L-, -(CH2) t1 -L-(CH2) t2 -, -C(=O)-, O, S, or -NH-;

[0106] Y 5 is -CH-, or N;

[0107] Y 6 is O, or -NH-;

[0108] Y 7 is O, or -NH-;

[0109] L is -C(=O)-, O, S, or -NH-;

[0110] t1, t2 and t3, at each occurrence, are independently 1, 2, 3 or 4;

[0111] t4 is 1; t5 is 1 or 2; and

[0112] represents a single bond or a double bond; and

[0113] Where R 1 Optionally, 0, 1, 2, 3 or 4 R 5 replace;

[0114] R 2 Each occurrence is independently H, D, F, Cl, Br, I, -OH, -CN, -NO2, -NR e R f , C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy, C 1-6 Hydroxyalkyl, C 1-6 Aminoalkyl, C 1-6 Cyanoalkyl, C 2-9 Heterocyclic group, C 3-8 Cycloalkyl, C 6-10 Aryl, or C 1-9 Heteroaryl; wherein R 2 At each occurrence, 0, 1, 2, 3, 4 or 5 independently selected from H, D, oxo (=O), F, Cl, Br, I, -OH, -NH2, -CN, -NO2, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Alkoxy and C 1-6 haloalkyl radical substitution;

[0115] R 3 It is H, D, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Haloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-8 Cycloalkyl, C 3-8 Deuterated cycloalkyl, C 2-9 Heterocyclic group, C 6-10 Aryl, or C 1-9 Heteroaryl; wherein R3 Optionally, 0, 1, 2, 3, 4 or 5 R 6 replace;

[0116] R 4 Each occurrence is independently H, D, F, Cl, Br, I, -OH, -CN, -NO2, -NR e R f , C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy, C 1-6 Hydroxyalkyl, C 1-6 Aminoalkyl, C 1-6 Cyanoalkyl, C 2-9 Heterocyclic group, C 3-8 Cycloalkyl, C 6-10 Aryl, or C 1-9 Heteroaryl; wherein R 4 At each occurrence, 0, 1, 2, 3, 4 or 5 independently selected from H, D, oxo (=O), F, Cl, Br, I, -OH, -NH2, -CN, -NO2, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Alkoxy and C 1-6 haloalkyl radical substitution;

[0117] R 5 and R 6 , each occurrence is independently H, D, oxo (=O), F, Cl, Br, I, -OH, -CN, -NO2, -NR e R f 、-C(=O)R 7 , C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy, C 1-6 Hydroxyalkyl, C 1-6 Aminoalkyl, C 1-6 Cyanoalkyl, C 2-9 Heterocyclic group, C 3-8 Cycloalkyl, C 6-10 Aryl, or C 1-9 heteroaryl; wherein each -NRe R f 、-C(=O)R 7 , C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy, C 1-6 Hydroxyalkyl, C 1-6 Aminoalkyl, C 1-6 Cyanoalkyl, C 2-9 Heterocyclic group, C 3-8 Cycloalkyl, C 6-10 Aryl and C 1-9 Heteroaryl is optionally substituted by 0, 1, 2, 3, 4 or 5 groups independently selected from H, D, oxo (=O), F, Cl, Br, I, -OH, -NH2, -CN, -NO2, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Alkoxy and C 1-6 haloalkyl radical substitution;

[0118] R 5a Yes-C(=O)R 7 , C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy, C 1-6 Hydroxyalkyl, C 1-6 Aminoalkyl, C 1-6 Cyanoalkyl, C 2-9 Heterocyclic group, C 3-8 Cycloalkyl, C 6-10 Aryl, or C 1-9 heteroaryl; wherein each -C(=O)R 7 , C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy, C 1-6 Hydroxyalkyl, C 1-6 Aminoalkyl, C 1-6 Cyanoalkyl, C 2-9 Heterocyclic group, C 3-8Cycloalkyl, C 6-10 Aryl and C 1-9 Heteroaryl is optionally substituted by 0, 1, 2, 3, 4 or 5 groups independently selected from H, D, oxo (=O), F, Cl, Br, I, -OH, -NH2, -CN, -NO2, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Alkoxy and C 1-6 haloalkyl radical substitution;

[0119] R e , R f , R 7 and R 7a , at each occurrence, are independently H, D, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl, C 3-8 Cycloalkyl, C 3-8 Cycloalkyl C 1-6 Alkyl, C 2-9 Heterocyclic group, C 2-9 Heterocyclyl C 1-6 Alkyl, C 6-10 Aryl, C 6-10 Aryl C 1-6 Alkyl, C 1-9 Heteroaryl, or C 1-9 Heteroaryl C 1-6 Alkyl; wherein each C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl, C 3-8 Cycloalkyl, C 3-8 Cycloalkyl C 1-6 Alkyl, C 2-9 Heterocyclic group, C 2-9 Heterocyclyl C 1-6 Alkyl, C 6-12 Aryl, C 6-12 Aryl C 1-6 Alkyl, C 1-9 Heteroaryl and C 1-9 Heteroaryl C 1-6 The alkyl radical is optionally substituted with 0, 1, 2, 3, 4 or 5 radicals independently selected from H, D, oxo (=O), F, Cl, Br, I, -OH, -NH2, -CN, -NO2, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Alkoxy and C 1-6 haloalkyl radical substitution; and

[0120] n is 0, 1, 2, or 3.

[0121] In some embodiments of Formula (II), W is

[0122] C 3-8 Cycloalkyl, C 3-8 Heterocyclic group,

[0123] Where Z 1 , Z 2 and Z 3 are each independently -CH-, or N; and

[0124] wherein W is optionally replaced by 0, 1, 2, 3 or 4 R 4 replace.

[0125] In some embodiments, the compound represented by formula (II) has the structure represented by formula (IIa):

[0126]

[0127] or a stereoisomer, tautomer, deuterated form, nitrogen oxide, solvate, or pharmaceutically acceptable salt thereof;

[0128] Wherein, m is 0, 1, 2 or 3.

[0129] In some embodiments of Formula (II) or (IIa), R 1 yes Where R 1 Optionally 0, 1, 2, 3 or 4 R 5 replace.

[0130] In some embodiments of Formula (II) or (IIa), R 4 Each occurrence is independently H, D, F, Cl, Br, I, -OH, -CN, -NO2, -NH2, C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Haloalkyl, C 1-3 Halogenated alkoxy, C 1-3 Hydroxyalkyl, C 1-3 Aminoalkyl, C 1-3 Cyanoalkyl, C 3-6 Heterocyclic group, C 3-6 Cycloalkyl, phenyl, or a heteroaryl group consisting of 5-6 atoms; wherein R 4At each occurrence, each is independently optionally replaced by 0, 1, 2, 3, 4 or 5 independently selected from H, D, oxo (=O), F, Cl, Br, I, -OH, -NH2, -CN, -NO2, C 1-3 Alkyl, C 1-3 Deuterated alkyl, C 1-3 Alkoxy and C 1-3 The haloalkyl group is substituted.

[0131] In some embodiments of Formula (II) or (IIa), R 4 At each occurrence, each is independently R 4 Each occurrence is independently H, D, F, Cl, Br, I, -OH, -CN, -NO2, -NH2, methyl, ethyl, isopropyl, methoxy, ethoxy, fluoromethyl, -CD3, -CHD2, -CH2D, cyclopropyl, phenyl, or a heteroaryl group consisting of 5-6 atoms; wherein each R 4 , independently and optionally replaced by 0, 1, 2, 3, 4 or 5 independently selected from H, D, oxo (=O), F, Cl, Br, I, -OH, -NH2, -CN, -NO2, C 1-3 Alkyl, C 1-3 Deuterated alkyl, C 1-3 Alkoxy and C 1-3 The haloalkyl group is substituted.

[0132] In some embodiments of Formula (II) or (IIa), R 5a Yes-C(=O)R 7 , C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-4 Halogenated alkoxy, C 1-4 Hydroxyalkyl, C 1-4 Aminoalkyl, C 1-4 Cyanoalkyl, C 3-6 Heterocyclic group, C 3-6 Cycloalkyl, C 6-10 Aryl, or C 1-9 heteroaryl; wherein each -C(=O)R 7 , C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-4 Halogenated alkoxy, C1-4 Hydroxyalkyl, C 1-4 Aminoalkyl, C 1-4 Cyanoalkyl, C 3-6 Heterocyclic group, C 3-6 Cycloalkyl, C 6-10 Aryl and C 1-9 Heteroaryl is optionally substituted by 0, 1, 2, 3, 4 or 5 groups independently selected from H, D, oxo (=O), F, Cl, Br, I, -OH, -NH2, -CN, -NO2, C 1-6 Alkyl, C 1-4 Deuterated alkyl, C 1-4 Alkoxy and C 1-4 The haloalkyl group is substituted.

[0133] In some embodiments of Formula (II) or (IIa), R 5a It is methyl, ethyl, isopropyl, cyclopropyl, -CF3, -CH2CH2OH, -C(=O)CH3, -C(=O)CH2CH3, or -CH2CH2OCH3.

[0134] In some embodiments of Formula (I), (Ia), (Ib), (II) or (IIa), R 3 is cyclopropyl, pyridyl, or phenyl; wherein R 3 Optionally, 0, 1, 2, 3, 4 or 5 independently selected from H, D, F, Cl, Br, I, -OH, -CN, -NO2, -NR e R f , C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Deuterated alkyl and C 1-3 The haloalkyl group is substituted.

[0135] In some embodiments of Formula (I), (Ia), (Ib), (II) or (IIa), R 5 and R 6 Each occurrence is independently H, D, oxo (=O), F, Cl, Br, I, -OH, -CN, -NO2, -NR e R f 、-C(=O)R 7 , C 1-3 Alkyl, C 1-6 Deuterated alkyl, C 1-3 Alkoxy, C 1-3 Haloalkyl, C 1-3 Hydroxyalkyl, C 3-6 Cycloalkyl, phenyl, or a heteroaryl group consisting of 5-6 atoms; wherein each -NR e R f、-C(=O)R 7 , C 1-3 Alkyl, C 1-6 Deuterated alkyl, C 1-3 Alkoxy, C 1-3 Haloalkyl, C 1-3 Hydroxyalkyl, C 3-6 Cycloalkyl, phenyl and 5-6 atoms of heteroaryl are independently optionally substituted by 0, 1, 2, 3, 4 or 5 atoms independently selected from H, D, oxo (=O), F, Cl, Br, I, -OH, -NH2, -CN, -NO2, C 1-3 Alkyl, C 1-3 Deuterated alkyl, C 1-3 Alkoxy and C 1-3 The haloalkyl group is substituted.

[0136] In some embodiments of Formula (I), (Ia), (Ib), (II) or (IIa), R 5 and R 6 Each occurrence is independently H, D, F, Cl, Br, I, -OH, -CN, -NO2, -NR e R f , methyl, ethyl, isopropyl, cyclopropyl, -CD3, -OCD3, -CF3, -CH2CH2OH, -C(=O)CH3, -C(=O)CH2CH3, or -CH2CH2OCH3.

[0137] In some embodiments of Formula (I), (Ia), (II) or (IIa), R a , R b and R c Each independently represents H, D, F, -CN, -NO2, -NR e R f , methyl, ethyl, isopropyl, -CD3, -CHD2, -CH2D, methoxy, ethoxy, halomethyl, haloethyl, or -CH2CH2OH.

[0138] In some embodiments of Formula (I), (Ia), (Ib), (II) or (IIa), R e , R f , R 7 and R 7a , at each occurrence, are independently H, D, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 1-4 Haloalkyl, C 1-4 Hydroxyalkyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkyl C 1-4 Alkyl, C3-6 Heterocyclic group, C 3-6 Heterocyclyl C 1-4 Alkyl, C 6-10 Aryl, C 6-10 Aryl C 1-4 Alkyl, C 1-9 Heteroaryl, or C 1-9 Heteroaryl C 1-4 Alkyl; wherein each C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 1-4 Haloalkyl, C 1-4 Hydroxyalkyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkyl C 1-4 Alkyl, C 3-6 Heterocyclic group, C 3-6 Heterocyclyl C 1-4 Alkyl, C 6-10 Aryl, C 6-10 Aryl C 1-4 Alkyl, C 1-9 Heteroaryl and C 1-9 Heteroaryl C 1-4 The alkyl group is optionally substituted with 0, 1, 2, 3, 4 or 5 independently selected from H, D, oxo (=O), F, Cl, Br, I, -OH, -NH2, -CN, -NO2, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 1-4 Alkoxy and C 1-4 The haloalkyl group is substituted.

[0139] In some embodiments of Formula (I), (Ia), (Ib), (II) or (IIa), R e , R f , R 7 and R 7a , at each occurrence, are independently H, D, C 1-2 Alkyl, C 1-2 Deuterated alkyl, C 1-2 haloalkyl, phenyl, or cyclopropyl.

[0140] In some embodiments of Formula (I), (Ia) or (Ib), the compound is a compound having one of the following structures:

[0141]

[0142]

[0143]

[0144] or a stereoisomer, tautomer, deuterated substance, nitrogen oxide, solvate, or pharmaceutically acceptable salt thereof.

[0145] In some embodiments of formula (II) or (IIa), the compound is a compound having one of the following structures:

[0146]

[0147]

[0148] or a stereoisomer, tautomer, nitrogen oxide, solvate, or pharmaceutically acceptable salt thereof.

[0149] Unless otherwise indicated, stereoisomers, tautomers, solvates, metabolites or pharmaceutically acceptable salts of the compounds represented by formula (I), (Ia), (Ib), (II) or (IIa) are all included in the scope of the present invention.

[0150] The compounds disclosed herein may contain asymmetric or chiral centers and may therefore exist in different stereoisomeric forms. The present invention is intended to include all stereoisomeric forms of the compounds of formula (I), (Ia), (Ib), (II) or (IIa), including but not limited to diastereomers, enantiomers, atropisomers and geometric (or conformational) isomers, and mixtures thereof such as racemic mixtures, as part of the present invention.

[0151] In structures disclosed herein, when the stereochemistry of any particular chiral atom is not indicated, all stereoisomers of the structure are contemplated within the present invention and are included as compounds disclosed herein. When stereochemistry is indicated by a solid wedge or dashed line representing a particular configuration, the stereoisomers of the structure are thus unambiguous and defined.

[0152] The compound shown in formula (I), (Ia), (Ib), (II) or (IIa) can exist in the form of a salt. In one embodiment, the salt refers to a pharmaceutically acceptable salt. The term "pharmaceutically acceptable" means that the substance or composition must be chemically and / or toxicologically compatible with the other ingredients of the formulation and / or the mammal treated therewith. In another embodiment, the salt is not necessarily a pharmaceutically acceptable salt, and can be an intermediate for preparing and / or purifying the compound shown in formula (I), (Ia), (Ib), (II) or (IIa) and / or for separating the enantiomer of the compound shown in the present formula (I), (Ia), (II) or (IIa).

[0153] In another aspect, the present invention relates to an intermediate for preparing a compound represented by formula (I), (Ia), (Ib), (II) or (IIa).

[0154] On the other hand, the present invention relates to methods for preparing, separating and purifying compounds represented by formula (I), (Ia), (Ib), (II) or (IIa).

[0155] On the other hand, the present invention provides a pharmaceutical composition comprising a compound of the present invention. In one embodiment, the pharmaceutical composition of the present invention further comprises a pharmaceutically acceptable excipient, diluent or carrier, or a combination thereof. In another embodiment, the pharmaceutical composition can be a liquid, solid, semi-solid, gel or spray formulation.

[0156] In some embodiments, the pharmaceutical composition described herein further comprises an additional therapeutic agent.

[0157] On the other hand, the present invention relates to a use of the compound described in the present invention or the pharmaceutical composition described in the present invention in the preparation of a medicament for preventing, treating, curing, and / or alleviating PI3-kinase-mediated diseases, disorders, and / or conditions, or inhibiting PI3-kinase activity.

[0158] In some embodiments, the PI3-kinase mediated disease, disorder, and / or condition is selected from respiratory disease, viral infection, non-viral respiratory infection, allergic disease, autoimmune disease, inflammatory disease, cardiovascular disease, hematological malignancy, neurodegenerative disease, pancreatitis, multiple organ failure, kidney disease, platelet aggregation, cancer, sperm motility, transplant rejection, graft rejection, lung injury or pain.

[0159] In some embodiments, the PI3-kinase mediated disease, disorder, and / or condition is selected from asthma, chronic obstructive pulmonary disease (COPD), viral respiratory infection, viral respiratory disease exacerbation, aspergillosis, leishmaniasis, allergic rhinitis, allergic dermatitis, rheumatoid arthritis, multiple sclerosis, inflammatory bowel disease, thrombosis, atherosclerosis, hematological malignancies, neurodegenerative diseases, pancreatitis, multiple organ failure, kidney disease, platelet aggregation, cancer, sperm motility, transplant rejection, graft rejection, lung injury, pain associated with rheumatoid arthritis or osteoarthritis, back pain, systemic inflammatory pain, retrohepatic neuralgia, diabetic neuropathy, inflammatory neuropathic pain (trauma), trigeminal neuralgia, or central pain.

[0160] In some embodiments, the cancer is selected from acute myeloid leukemia, myelodysplastic syndrome, myeloproliferative disorder, chronic myeloid leukemia, T-cell acute lymphoblastic leukemia, B-cell acute lymphoblastic leukemia, non-Hodgkin lymphoma, B-cell lymphoma, solid tumor, or breast cancer.

[0161] In some embodiments, wherein the PI3-kinase is PI3K-γ.

[0162] Pharmaceutical compositions, preparations and administration of the compounds of the present invention

[0163] The present invention provides a pharmaceutical composition, which comprises a compound disclosed in the present invention, or a compound listed in the embodiments, or a stereoisomer, tautomer, nitrogen oxide, solvate, metabolite or pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable excipient, diluent, carrier, solvent or a combination thereof. The amount of the compound in the pharmaceutical composition disclosed in the present invention refers to an amount that can effectively detect the inhibition of protein kinase in a biological sample or a patient.

[0164] It should also be recognized that certain compounds of the present invention may be present in free form for treatment, or, if appropriate, in the form of pharmaceutically acceptable derivatives thereof. Some non-limiting embodiments of pharmaceutically acceptable derivatives include pharmaceutically acceptable salts, esters, salts of these esters, or any other adducts or derivatives that can directly or indirectly provide the compounds of the present invention or their metabolites or residues when administered to a patient in need thereof.

[0165] In document such as Remington: The Science and Practice of Pharmacy, 21st edition, 2005, ed. DB Troy, Lippincott Williams & Wilkins, Philadelphia, and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and JC Boylan, 1988-1999, Marcel Dekker, New York, various carriers for configuring pharmaceutically acceptable compositions, and known techniques for their preparation are disclosed, and the respective contents of these documents are incorporated by reference into the present invention. Except any commonly used carriers such as those which are incompatible with the disclosed compounds of the present invention due to any undesirable biological effects or any other components in the pharmaceutically acceptable compositions interacting in a harmful manner, it is concerned that their application belongs to the scope of the present invention.

[0166] The pharmaceutical composition provided by the present invention can be co-formulated with other active ingredients that do not impair the intended therapeutic effect, or co-formulated with substances that supplement the intended effect.

[0167] Uses of the compounds and compositions of the present invention

[0168] The compounds of the present invention are inhibitors of kinase activity, in particular inhibitors of PI3-kinase activity. Compounds that are PI3-kinase inhibitors can be used to treat disorders in which the underlying pathology (at least in part) is attributed to improper PI3-kinase activity, such as asthma, chronic obstructive pulmonary disease (COPD), viral infections, non-viral respiratory infections, allergic diseases, autoimmune diseases, inflammatory diseases, cardiovascular diseases, malignant blood diseases, neurodegenerative diseases, pancreatitis, multiple organ failure, kidney disease, platelet aggregation, cancer, sperm motility, transplant rejection, graft rejection, lung injury or pain, etc. "Improper PI3-kinase activity" refers to any PI3-kinase activity that deviates from the normal PI3-kinase activity expected in a specific patient. Improper PI3-kinase can take the form of, for example, an abnormal increase in activity, or a distorted or dysregulated form of PI3-kinase. These improper activities can be caused by, for example, overexpression or mutation of protein kinases that are improperly or uncontrolled and activated. Therefore, in another aspect, the present invention relates to a method for treating the disease or condition.

[0169] Such diseases or conditions include, but are not limited to, respiratory diseases, including asthma, chronic obstructive pulmonary disease, and idiopathic pulmonary fibrosis (IPF); viral infections, including viral respiratory infections and exacerbations of viral respiratory diseases, such as asthma and COPD; non-viral respiratory infections, including aspergillosis and leishmaniasis; allergic diseases, including allergic rhinitis and atopic dermatitis; autoimmune diseases, including rheumatoid arthritis and multiple sclerosis; inflammatory diseases, including inflammatory bowel disease; cardiovascular diseases, including thrombosis and atherosclerosis; hematological malignancies; neurodegenerative diseases; pancreatitis; multiple organ failure; kidney disease; platelet aggregation; cancer; sperm motility; transplant rejection; graft rejection; lung injury; and pain, including pain associated with rheumatoid arthritis or osteoarthritis, back pain, systemic inflammatory pain, retrohepatic neuralgia, diabetic neuropathy, inflammatory neuropathic pain (trauma), trigeminal neuralgia, and central pain. In one embodiment, such disorders include respiratory diseases, including asthma and chronic obstructive pulmonary disease (COPD); allergic diseases, including allergic rhinitis and atopic dermatitis; autoimmune diseases, including rheumatoid arthritis and multiple sclerosis; inflammatory diseases, including inflammatory bowel disease; cardiovascular diseases, including thrombosis and atherosclerosis; hematological malignancies; neurodegenerative diseases; pancreatitis; multiple organ failure; kidney disease; platelet aggregation; cancer; sperm motility; transplant rejection; graft rejection; lung injury; and pain, including pain associated with rheumatoid arthritis or osteoarthritis, back pain, systemic inflammatory pain, retrohepatic neuralgia, diabetic neuropathy, inflammatory neuropathic pain (traumatic), trigeminal neuralgia, and central pain.

[0170] In such diseases or disorders, the cancer is selected from acute myeloid leukemia, myelodysplastic syndrome, myeloproliferative disease, chronic myeloid leukemia, T-cell acute lymphoblastic leukemia, B-cell acute lymphoblastic leukemia, non-Hodgkin lymphoma, B-cell lymphoma, solid tumor, or breast cancer.

[0171] The therapeutic method of the present invention comprises administering a safe and effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof to a patient in need thereof. Various embodiments of the present invention include methods for treating any of the disorders or diseases mentioned in the present invention by administering a safe and effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof to a patient in need thereof.

[0172] Combination therapy

[0173] The compounds of the invention may be administered as the sole active agent, or may be administered in combination with other therapeutic agents, including other compounds that have the same or similar therapeutic activity and that are determined to be safe and effective for such combination administration.

[0174] In one aspect, the present invention provides a method for treating, preventing or ameliorating a disease or condition, comprising administering a safe and effective amount of a combination drug comprising a compound disclosed herein and one or more therapeutically active agents. In some embodiments, the combination drug comprises one or two other therapeutic agents.

[0175] Examples of other therapeutic agents include, but are not limited to: anti-cancer agents, including chemotherapeutic agents and anti-proliferative agents; anti-inflammatory agents; and immunomodulatory or immunosuppressive agents.

[0176] On the other hand, the present invention provides products including compounds of the present invention and at least one other therapeutic agent, which can be prepared as a combination for simultaneous, separate or sequential administration in treatment. In some embodiments, the treatment is for the treatment of diseases or symptoms mediated by one or more protein kinases, such as PI3k-kinase activity. The products provided by the combined preparation include compositions present in the same pharmaceutical composition and containing compounds disclosed herein and other therapeutic agents, or compounds disclosed herein and other therapeutic agents in different forms, for example, a kit.

[0177] In another aspect, the present invention provides a pharmaceutical composition comprising a compound disclosed herein and one or more additional therapeutic agents. In some embodiments, the pharmaceutical composition may comprise a pharmaceutically acceptable excipient as described above.

[0178] On the other hand, the present invention provides a medicine box comprising two or more separate pharmaceutical compositions, wherein at least one pharmaceutical composition comprises a disclosed compound of the present invention. In some embodiments, the medicine box includes a tool for separately holding the composition, such as a container, a separate bottle or a separate foil box. An example of such a medicine box is a blister package, which is commonly used for packaging tablets, capsules, etc.

[0179] The compounds disclosed herein can be administered as the sole active ingredient or co-administered with other therapeutic agents, for example, as an adjuvant.

[0180] In some embodiments, the other therapeutic agents include, for example, immunosuppressants, immunomodulators or other anti-inflammatory agents, drugs for treating or preventing acute or chronic rejection of allogeneic or xenogeneic transplants, or inflammation, or autoimmune diseases, or chemotherapeutic agents, such as malignant cell antiproliferative agents.

[0181] The compound represented by formula (I) of the present invention is used in combination with other immunosuppressants / immunomodulators, anti-inflammatory agents, chemotherapeutic agents or anti-infective agents, wherein the dosage of the immunosuppressants / immunomodulators, anti-inflammatory agents, chemotherapeutic agents or anti-infective agents used in combination depends on the type of combined medication, whether it is a steroid compound or a calcineurin inhibitor, as well as the specific drugs being used for treatment and the treatment conditions, etc.

[0182] Examples of anti-inflammatory agents include nonsteroidal anti-inflammatory drugs (NSAIDs). Examples of NSAIDs include sodium cromoglycate, nedocromil sodium, phosphodiesterase (PDE) inhibitors (e.g., theophylline, PDE4 inhibitors, or mixed PDE3 / PDE4 inhibitors), leukotriene antagonists, leukotriene synthesis inhibitors (e.g., montelukast), iNOS inhibitors, trypsin and elastase inhibitors, beta-2 integrin antagonists and adenosine receptor agonists or antagonists (e.g., adenosine 2α receptor agonists), cytokine antagonists (e.g., chemokine receptor antagonists, including CCR3 antagonists), cytokine synthesis inhibitors, or 5-lipoxygenase inhibitors.

[0183] The compounds of formula (I) may also be advantageously used in combination with other compounds or in combination with other therapeutic agents, especially antiproliferative agents. Such antiproliferative agents include, but are not limited to, aromatase inhibitors; antiestrogens; topoisomerase I inhibitors; topoisomerase II inhibitors; microtubule active agents; alkylating agents; histone deacetylase inhibitors; compounds that induce cell differentiation processes; cyclooxygenase inhibitors; MMP inhibitors; mTOR inhibitors; antitumor antimetabolites; platinum compounds; compounds that target / reduce protein or lipid kinase activity and other antiangiogenic compounds; compounds that target, reduce or inhibit protein or lipid phosphatase activity; gonadorelin agonists; antiandrogens; methionine aminopeptidase inhibitors; bisphosphonates; biological response modifiers; antiproliferative antibodies; heparanase inhibitors; inhibitors of Ras oncogenic isoforms; telomerase inhibitors; proteasome inhibitors; agents for treating hematological tumors; compounds that target, reduce or inhibit Flt-3 activity; Hsp90 inhibitors; temozolomide and folinic acid.

[0184] "Combination" means a fixed combination in a single dosage unit form or a kit of parts for combined administration, wherein the compounds disclosed herein and the combination partner can be administered independently at the same time or can be administered separately within a certain time interval, especially so that the combination partner exhibits a cooperative, such as synergistic effect. As used herein, the term "co-administration" or "combined administration" and the like are intended to encompass administration of the selected combination partner to a single individual (e.g., patient) in need thereof, and are intended to include treatment regimens in which the substances are not necessarily administered by the same route of administration or at the same time.

[0185] Treatment

[0186] In some embodiments, the treatment methods disclosed herein include administering a safe and effective amount of a compound of the present invention or a pharmaceutical composition comprising a compound of the present invention to a patient in need thereof. The various embodiments disclosed herein include methods for treating the diseases or conditions described herein by administering a safe and effective amount of a compound of the present invention or a pharmaceutical composition comprising a compound of the present invention to a patient in need thereof.

[0187] In some embodiments, the compounds disclosed in the present invention or the pharmaceutical composition comprising the compounds disclosed in the present invention can be administered once, or according to the dosing regimen, several times at different time intervals within a specified time period. For example, once, twice, three times or four times a day. In one embodiment, it is administered once a day. In another embodiment, it is administered twice a day. It can be administered until the desired therapeutic effect is achieved or the desired therapeutic effect is maintained indefinitely. The suitable dosing regimen of the compounds disclosed in the present invention or the pharmaceutical composition comprising the compounds disclosed in the present invention depends on the pharmacokinetic properties of the compounds, such as dilution, distribution and half-life, which can be determined by technicians. In addition, the suitable dosing regimen of the compounds disclosed in the present invention or the pharmaceutical composition comprising the compounds disclosed in the present invention, including the duration of the implementation of the regimen, depends on the disease being treated, the severity of the disease being treated, the age and physical condition of the patient being treated, the medical history of the patient being treated, the nature of the concurrent therapy, the desired therapeutic effect, and other factors within the knowledge and experience of the technician. The technical personnel in the field should also understand that the appropriate dosing regimen may be required to be adjusted for the response of individual patients to the dosing regimen, or when the needs of individual patients change over time.

[0188] The pharmaceutical composition or combination / combination of the present invention can be about 1-1000mg active ingredient, or about 1-500mg or about 1-250mg or about 1-150mg or about 0.5-100mg, or about 1-50mg of active ingredient unit dose for individuals of about 50-70kg. The therapeutically effective dose of the compound, pharmaceutical composition or combination thereof depends on the species, body weight, age and individual disease, disorder or condition or severity to be treated of the individual. A physician of ordinary skill in the art, a clinician or a veterinarian can easily determine the effective amount of each active ingredient for preventing, treating or inhibiting the progression of a disease or disorder. The above-cited dosage characteristics have been confirmed in in vitro and in vivo tests using favorable mammals, such as mice, rats, dogs, monkeys, or isolated organs, tissues and specimens thereof. The compounds of the present invention can be used in vitro in the form of a solution, such as an aqueous solution, and can also be used in vivo in the form of a suspension or an aqueous solution, enterally, parenterally and through a suitable vein. The range of therapeutically effective amounts in vivo depends on the route of administration, and is between about 0.01-500 mg / kg, or between about 1-100 mg / kg.

[0189] The compounds disclosed in the present invention can be administered simultaneously with, before or after, one or more other therapeutic agents. The compounds disclosed in the present invention can be administered separately with other therapeutic agents through the same or different administration routes, or in the form of the same pharmaceutical composition.

[0190] General synthetic scheme

[0191] To describe the present invention, the following examples are listed. However, it should be understood that the present invention is not limited to these examples, which are only provided to provide methods for practicing the present invention.

[0192] Generally, the compounds of the present invention can be prepared by the methods described herein, unless otherwise specified, wherein the substituents are defined as shown in formula (I), (Ia), (Ib), (II) or (IIa). The following reaction schemes and examples are provided to further illustrate the present invention.

[0193] In the examples described below, all temperatures are set in degrees Celsius unless otherwise indicated. Reagents were purchased from commercial suppliers such as Aldrich Chemical Company, Alfa Chemical Company, Shanghai Shaoyuan Reagent Co., Ltd., SAIN Chemical Technology (Shanghai) Co., Ltd., Shanghai Baide Pharmaceutical Technology Co., Ltd., and Shanghai Haohong Biomedical Technology Co., Ltd. Unless otherwise specified, the reagents used in the present invention can be used without further purification. Common solvents were purchased from commercial suppliers such as Beijing Haizhiyuan Weiye Technology Co., Ltd.

[0194] Anhydrous THF, dioxane, DCM, toluene and DMF were purchased from commercial suppliers, such as Energychemical Company and Aldrich Chemical Company. EtOAc, PE, CH3CN, NMP and DMSO were treated with anhydrous Na2SO4 before use.

[0195] The following reactions were generally carried out under positive pressure of nitrogen or argon or with a drying tube over anhydrous solvents (unless otherwise indicated), reaction bottles were plugged with appropriate rubber stoppers, and substrates were injected via syringes. All glassware was dried.

[0196] The chromatographic column used was a silica gel column, and the silica gel (300-400 mesh) was purchased from Qingdao Ocean Chemical Plant.

[0197] 1 H NMR spectra and 13 The C / 2D data were collected on a Bruker Avance III 400 MHz. 1H NMR spectra were obtained in CDC13, DMSO-d6, CD3OD or acetone-d6 as solvents (in ppm) and TMS (0 ppm) or chloroform (7.26 ppm) as reference standards. When multiple peaks are present, the following abbreviations are used: s (singlet), d (doublet), t (triplet), m (multiplet), br (broadened), brs (broadened singlet), dd (doublet of doublets), dt (doublet of triplets). Coupling constants are expressed in Hertz (Hz).

[0198] LC / MS was performed on an Agilent 1260 (binary pump / DAD detector) coupled to an Agilent 6120 / 6125 mass spectrometer.

[0199] Method 1:

[0200] Column: HALO C18 2.7 μm, 4.6 mm×30 mm, mobile phase: MeCN (0.05% HCOOH)-Water (0.05% HCOOH); gradient: from 5% to 95% MeCN, elution 0.8 min, hold 0.8 min, total run time is 2.0 min; flow rate: 1.8 mL / min; column temperature: 45°C;

[0201] Method 2:

[0202] Column: HALO C18 2.7 μm, 4.6 mm×50 mm, mobile phase: MeCN (0.025% trifluoroacetic acid)-water (0.025% trifluoroacetic acid); gradient: from 5% to 95% MeCN, elution 1.0 min, hold 1.0 min, total run time is 2.5 min; flow rate: 1.8 mL / min; column temperature: 45°C.

[0203] Purity test by RP-HPLC:

[0204] Compound purity test on RP-HPLC (Shimadzu 2010 / 2030)

[0205] Method 1:

[0206] Column: Gemini 4.6×150mm 5um; Mobile phase: H2O (0.05% trifluoroacetic acid)-MeCN (0.05% trifluoroacetic acid). Gradient: from 10% to 100% MeCN, elution for 8 minutes, hold for 2 minutes. Flow rate: 1.2mL / min, column temperature: 35℃ / 40℃. Method 2:

[0207] Column: XBRIDGE 2.1×50mm, 3.5um; Mobile phase: H2O (0.05% trifluoroacetic acid)-MeCN (0.05% trifluoroacetic acid). Gradient: from 10% to 100% MeCN, elution for 7 minutes, hold for 1 minute. Flow rate: 0.8mL / min, column temperature: 35℃ / 40℃. Compound purification by SFC:

[0208] SFC purification was performed on a Thar P80 equipped with UV detection.

[0209] Method: Column CHIRALPAK AD-H 250 mm, 20 mm, 5 μm, modifier: 30% EtOH (0.2% NH4OH).

[0210] Compound purification by RP-HPLC:

[0211] RP-HPLC purification was performed on a Gilson purification system (322 or 306 pump and GX-281 fraction collector), Shimadzu LC20Ap, and Waters MS triggered purification system;

[0212] Method 1:

[0213] Columns Gemini C18 21x150 mm, 5μm Xbrige C18 19x150 mm, 5μm, Spolar C18 20x150mm and Ultimate AQ-C18 30x250 mm, 10μm

[0214] Mobile Phase:

[0215] 1. MeCN in water (0.1% HCOOH), flow rate: 20 ml / min, 50 ml / min, column 30x250 mm, 10 μm; wavelength: 210-400 nm. Inject the sample into DMSO (+optional formic acid and water), elute with a linear gradient from 10% to 95% MeCN over 10 minutes.

[0216] 2. MeCN in water (0.1% trifluoroacetic acid), flow rate: 20 ml / min, 50 ml / min, column 30x250 mm, 10 μm; wavelength: 210-400 nm. Samples were injected into DMSO (+optional formic acid and water), eluted with a linear gradient from 10% to 95% MeCN over 10 minutes.

[0217] 3. Aqueous solution of MeCN (0.1% NH3-H2O / 10mM NH4AC), flow rate: 20ml / min, 50ml / min, column 30x250 mm, 10μm; wavelength: 210-400nm. The sample was injected into DMSO (+optional formic acid and water), and eluted with a linear gradient from 10% to 95% MeCN for 10 minutes.

[0218] Typical synthetic steps for preparing the compounds disclosed in the present invention are shown in the following synthetic schemes. 1 , R 2 , R 3 , R 4a , R 4b , R 4c , R a , R b , X, Z 1 , Z 2 , W and n are all defined as described in the present invention.

[0219] Synthesis Scheme 1:

[0220]

[0221] The compound of the present invention having a structure as shown in formula (6) can be prepared by the general synthesis method described in Synthesis Scheme 1. The specific steps can be referred to in the Examples. In Synthesis Scheme 1, compound (1) can be obtained by purchasing commercial reagents or synthesizing with reference to literature (WO2016149160A1). Compound (1) and R 1 The substituted terminal alkyne is coupled under the catalysis of a suitable palladium salt or copper salt to obtain compound (3). The Boc protecting group is removed under acidic conditions to obtain a derivative of compound (4). Compound (4) and compound (5) react under suitable alkaline conditions or in the presence of a condensing agent to obtain the target kinase inhibitor (6).

[0222] Synthesis Scheme 2:

[0223]

[0224] The compound of the present invention having a structure as shown in formula (8) can be prepared by the general synthesis method described in Synthesis Scheme 2. For specific steps, refer to the Examples. In Synthesis Scheme 2, an amine derivative (4) and a carboxylic acid derivative (7) are condensed in the presence of a suitable condensing agent to obtain the target kinase inhibitor (8).

[0225] Fragment synthesis scheme 1:

[0226]

[0227] Alkyne derivatives having a structure as shown in formula (2) can be prepared by the general synthesis method described in fragment synthesis scheme 1. The specific steps can be referred to in the examples. In synthesis scheme 2, compound (2-a) is coupled with trimethylethynylsilane under suitable alkaline conditions (such as potassium carbonate, cesium carbonate and potassium phosphate, etc.) under the catalysis of suitable palladium salts and copper salts to obtain compound (2-b). TBAF is used to remove the TMS protecting group to obtain the acetylene derivative (2). Alternatively, R 1 The substituted aldehyde derivative (2-c) is reacted with dimethyl (1-diazo-2-oxopropyl)phosphonate at low temperature under appropriate alkaline conditions to obtain an acetylene derivative (2).

[0228] Fragment synthesis scheme 2:

[0229]

[0230] The carboxylic acid derivative having the structure shown in formula (5) can be prepared by the general synthesis method described in fragment synthesis scheme 2, and the specific steps can be referred to in the examples. In synthesis scheme 3, compound (5-a) is deethylated under suitable alkaline conditions to obtain a carboxylic acid derivative (5-b). Compound (5-b) and compound (4) are condensed in the presence of a condensing agent (such as EDCI or HATU) to obtain the target kinase inhibitor (6). Compound (5-b) and N-hydroxysuccinimide are condensed in the presence of a condensing agent to obtain compound (5-c). Under suitable alkaline conditions, compound (5-c) and compound (4) react to obtain the target kinase inhibitor (6).

[0231] Deuterated fragment example 1:

[0232] The deuterated building blocks used to synthesize the compounds of the present invention include commercial reagents and can be synthesized by reference documents (such as WO2017161116A1), specifically including but not limited to the following structures:

[0233] Example

[0234] Intermediate 1 2,5-dioxopyrrol-1-yl 2-aminopyrazolo[1,5-a]pyrimidine-3-carboxylate-5,7-D2

[0235]

[0236] Step 1) 2-aminopyrazolo[1,5-a]pyrimidine-3-carboxylic acid ethyl ester-5,7-D2

[0237] 1,1,3,3-tetraethoxypropane-1,3-D2 (1313.3 mg, 5.9 mmol) was added to a solution of 3,5-diamino-1H-pyrazole-4-carboxylic acid ethyl ester (1 g, 5.9 mmol) in acetic acid (30 mL). The mixture was heated to 120°C and stirred for 16 h, cooled to room temperature, concentrated under reduced pressure, and the residue was diluted with acetone (30 mL). After filtration, 2-aminopyrazolo[1,5-a]pyrimidine-3-carboxylic acid ethyl ester-5,7-D2 (1.2 g, 93.21% yield) was obtained. MS (ESI): 209.1 [M+H] + .

[0238] Step 2) 2-aminopyrazolo[1,5-a]pyrimidine-3-carboxylic acid-5,7-D2

[0239] LiOH (1.22 g, 29 mmol) was added to a solution of ethyl 2-aminopyrazolo[1,5-a]pyrimidine-3-carboxylate (1.2 g, 5.8 mmol) in MeOH / H2O (50 mL / 10 mL). The mixture was stirred at 60°C for 4 h, the solvent was removed under reduced pressure, and the residue was adjusted to pH 5 with 1N HCl. After suction filtration, the filter residue was dried to obtain 2-aminopyrazolo[1,5-a]pyrimidine-3-carboxylic acid-5,7-D2 (785 mg, 72.41% yield) as a yellow solid. MS (ESI): 181.0 [M+H] + .

[0240] Step 3) 2,5-dioxopyrrol-1-yl 2-aminopyrazolo[1,5-a]pyrimidine-3-carboxylate-5,7-D2

[0241] EDCI (279.8 mg, 1.46 mmol) and 1-hydroxypyrrolidine-2,5-dione (168 mg, 1.46 mmol) were added to a DMF (10 mL) solution of 2-aminopyrazolo[1,5-a]pyrimidine-3-carboxylic acid (200 mg, 1.12 mmol), and the mixture was stirred at room temperature for 16 h. After the reaction was completed, it was diluted with water (30 ml) and extracted three times with ethyl acetate (30 mL x 3). The combined organic phase was washed with saturated brine (40 mL x 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 2,5-dioxopyrrolidin-1-yl 2-aminopyrazolo[1,5-a]pyrimidine-3-carboxylate-5,7-D2 (238 mg, 69.32% yield) as a yellow solid. MS (ESI): 277.9 [M+H] + .

[0242] Intermediate 2(S)-3-(1-aminoethyl)-8-chloro-2-phenylisoquinolin-1(2H)-one

[0243]

[0244] Step 1) Tert-butyl (S)-(1-(methoxy(methyl)amino)-1-oxopropan-2-yl)carbamate

[0245] CDI (8.92 g, 0.055 mmol) was added to a solution of (tert-butoxycarbonyl)-L-alanine (8.0 g, 0.042 mol) in DCM (200 mL). The mixture was stirred at room temperature for 1 h, then N,O-dimethylhydroxylamine (4.95 g, 0.051 mol) and TEA (17.1 g, 0.17 mol) were added, and the mixture was stirred at 25 ° C for 16 h. The mixture was diluted with H2O (20 mL) and then extracted with DCM (50 mL×3). The combined organic layers were washed with brine (40 mL x 2), dried over Na2SO4, and concentrated under reduced pressure. The residue was purified by flash chromatography (DCM / MeOH=98 / 2) to give tert-butyl (S)-(1-(methoxy(methyl)amino)-1-oxopropane-2-yl)carbamate (6.8 g, 62.17% yield) as a white solid. MS(ESI):254.9[M+Na] + .

[0246] Step 2) 2-Chloro-6-methyl-N-phenylbenzamide

[0247] Oxalyl chloride (4.83 g, 0.038 mol) was added dropwise to a solution of 2-chloro-6-methylbenzoic acid (5.0 g, 0.029 mol) and DMF (210 mg, 0.29 mmol) in DCM (50 mL) at 0°C, and the mixture was stirred at 0°C for 2 h. The above reaction mixture was added to a solution of aniline (2.73 g, 0.029 mol) and TEA (8.88 g, 0.088 mol) in DCM (50 mL) at 0°C. The mixture was stirred at room temperature for 5 h. The mixture was diluted with H2O (30 ml) and extracted with DCM (50 mL×3). The combined organic layers were washed with brine (40 mL x 2), dried over Na2SO4, and concentrated under reduced pressure. The residue was purified by flash chromatography (PE / EA=3 / 1) to give 2-chloro-6-methyl-N-phenylbenzamide (5.66 g, 70.65% yield) as a yellow solid. MS (ESI): 245.9 [M+H] + .

[0248] Step 3) Tert-butyl (S)-(4-(3-chloro-2-(phenylcarbamoyl)phenyl)-3-oxobutan-2-yl)carbamate

[0249] At -30 ° C, under nitrogen, in 10 minutes, n-BuLi (4.3ml, 10.2mmol) solution was added dropwise to a stirred solution of 2-chloro-6-methyl-N-phenylbenzamide (1g, 4.1mmol) in THF (20mL), and the reaction mixture was stirred at -30 ° C for 1 hour. At -30 ° C and under nitrogen, in 10 minutes, i-PrMgCl (9.5ml, 12.3mmol) solution was added dropwise to a stirred solution of (S)-(1-(methoxy(methyl)amino)-1-oxopropane-2-yl)carbamic acid tert-butyl ester (1.43g, 6.15mmol) in THF (20mL), and the reaction mixture was stirred at -30 ° C for 1h. Then the resulting solution was slowly added to the above reaction mixture. The mixture was stirred at room temperature for 3 hours. The mixture was quenched with NH4Cl (30mL) and then extracted with EtOAc (30mL x 3). The combined organic layers were washed with brine (10 mL x 2), dried over Na2SO4, and concentrated under reduced pressure. The residue was purified by flash chromatography (PE / EA=3 / 1) to give (S)-tert-butyl (4-(3-chloro-2-(phenylcarbamoyl)phenyl)-3-oxobutan-2-yl)carbamate (1.45 g, 75.61% yield) as a yellow solid. MS (ESI): 438.8 [M+Na] + .

[0250] Step 4) (S)-3-(1-aminoethyl)-8-chloro-2-phenylisoquinolin-1(2H)-one

[0251] To a solution of (S)-tert-butyl (4-(3-chloro-2-(phenylcarbamoyl)phenyl)-3-oxobutan-2-yl)carbamate (1.4 g, 3.4 mmol) in MeOH (30 mL) was added concentrated hydrochloric acid (2.8 mL) at room temperature, and the mixture was stirred at 80°C for 2 hours. The mixture was concentrated under reduced pressure, and the residue was diluted with H2O (10 mL), and the pH was adjusted to 7-8 with NaHCO3 solution. The mixture was extracted with DCM (30 mL x 3), and the combined organic layers were washed with brine (20 mL x 2), dried over Na2SO4, and concentrated under reduced pressure. The residue was purified by flash chromatography (DCM / MeOH=95 / 5) to give (S)-3-(1-aminoethyl)-8-chloro-2-phenylisoquinolin-1(2H)-one (830 mg, 73.53% yield) as a yellow solid. MS(ESI):298.9[M+H] + .

[0252] Intermediate 3 2-amino-5-methylpyrazolo[1,5-a]pyrimidine-3-carboxylic acid

[0253]

[0254] Step 1) 2-amino-5-methylpyrazolo[1,5-a]pyrimidine-3-carboxylic acid ethyl ester

[0255] To a solution of ethyl 3,5-diamino-1H-pyrazole-4-carboxylate (1 g, 6.0 mmol) in DMF (10 mL) was added 4,4-dimethoxy-2-butanone (1.6 g, 12.0 mmol), the reaction was heated at 110 °C for 1 hour, then CH3COOH (361 mg, 6.0 mmol) was added and the reaction was heated for 2 hours. After cooling to room temperature, the mixture was neutralized with saturated sodium bicarbonate and diluted with water (30 mL). The mixture was extracted with EtOAc (40 mL x 3). The combined organic layers were washed with brine (60 mL x 2), dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by column chromatography (DCM / MeOH=20 / 1) to give ethyl 2-amino-5-methylpyrazolo[1,5-a]pyrimidine-3-carboxylate (940 mg, 71%) as a white solid. MS (ESI) m / z 224.1 [M+H] + .

[0256] Step 2) 2-amino-5-methylpyrazolo[1,5-a]pyrimidine-3-carboxylic acid

[0257] To a solution of ethyl 2-amino-5-methylpyrazolo[1,5-a]pyrimidine-3-carboxylate (940 mg, 4.3 mmol) in MeOH / H2O (20 mL / 4 mL) was added NaOH (854 mg, 21.4 mmol), and the mixture was stirred at 60°C for 16 hours. The mixture was concentrated under reduced pressure, and the residue was diluted with water (20 mL). The mixture was acidified to pH=3-4 with 1N HCl, and then the mixture was extracted with EtOAc (200 mL). The separated organic layer was washed with brine (100 mL), dried over Na2SO4, and concentrated in vacuo to give 2-amino-5-methylpyrazolo[1,5-a]pyrimidine-3-carboxylic acid (450 mg, 53%) as a yellow solid. MS (ESI) m / z 196.0 [M+H] + .

[0258] Example 1 (S)-2-amino-N-(1-(8-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-2-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-5,7-D2-3-carboxamide

[0259]

[0260] Step 1) Nitrosoproline

[0261] Concentrated hydrochloric acid (5 mL) was added to a solution of pyrrolidine-2-carboxylic acid and NaNO2 (4.19 g, 0.061 mmol) in water (10 mL) at 0°C. The mixture was stirred at room temperature for 16 h, diluted with water (20 mL), and extracted three times with ethyl acetate (30 mL x 3). The combined organic phase was washed with brine (40 mL x 2), dried over anhydrous sodium sulfate, and concentrated to remove the solvent to obtain nitrosoproline (5.5 g, 83% yield) as a white solid. MS (ESI): 145.1 [M+H] + . 1 H NMR (400MHz, DMSO) δ6.50 (d, J=8.6Hz, 1H), 5.81-5.67 (m, 1H), 5.07-4.90 (m, 2H), 4.64-4 .54(m,1H),3.44-3.37(m,1H),2.31-2.19(m,1H),2.07-1.96(m,1H),1.41-1.33(m,9H).

[0262] Step 2) 5,6-dihydro-4H-pyrrolo[1,2-c][1,2,3]oxadiazole-7-azonia-3-inium salt

[0263] At 0°C, TFAA (3.8 g, 0.018 mol) was added to a solution of nitrosoproline (2 g, 0.014 mol) in acetonitrile (6 mL) and stirring was continued for 3 h. The mixture was quenched with potassium carbonate (3.1 g, 0.022 mol) and diluted with water (20 mL). The mixture was extracted three times with ethyl acetate (20 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM / MeOH=20 / 1) to give 5,6-dihydro-4H-pyrrole[1,2-c][1,2,3]oxadiazole-7-azonia-3-inium salt (1.5 g, 77% yield) as a brown oil. MS (ESI): 252.8 [2M+H] + . 1 H NMR (400MHz, CDCl3) δ4.44-4.39(m,2H), 2.94-2.90(m,2H), 2.81-2.74(m,2H).

[0264] Step 3) 5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole-2-carboxylic acid ethyl ester

[0265] A xylene solution (10 mL) of 5,6-dihydro-4H-pyrrole [1,2-c] [1,2,3] oxadiazole-7-azonium-3-indole (0.5 g, 4 mmol) and methyl prop-2-ynoate (1.18 g, 12 mmol) was heated to 140 ° C and stirred for 16 h. Cool to room temperature and concentrate. The residue was diluted with EtOAc (30 mL) and washed with saturated brine (20 mL), and the organic phase was dried over saturated sodium sulfate and concentrated under reduced pressure. The resulting residue was separated by silica gel column chromatography to obtain 5,6-dihydro-4H-pyrrole [1,2-b] pyrazole-2-carboxylic acid ethyl ester (0.23 g, 30% yield) as a brown oil. MS (ESI): 181.1 [M + H] + . 1 H NMR (400MHz, CDCl3) δ6.55 (s, 1H), 4.39 (q, J = 7.2Hz, 2H), 4.25-4.16 (m, 2H), 2.99-2.87 (m, 2H), 2.69-2.56 (m, 2H), 1.40 (t, J = 7.2Hz, 3H).

[0266] Step 4) (5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-2-yl)methanol

[0267] At 0°C, LiAlH4 (48 mg, 1.27 mmol) was added to a tetrahydrofuran solution (10 mL) of 5,6-dihydro-4H-pyrrole [1,2-b] pyrazole-2-carboxylic acid ethyl ester (230 mg, 1.27 mmol), and the mixture was stirred at 0°C for 2 h. The mixture was quenched with a 10% aqueous solution of NaOH (2 mL) and filtered. The filtrate was extracted three times with ethyl acetate (20 mL x 3), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to remove the solvent to obtain (5,6-dihydro-4H-pyrrole [1,2-b] pyrazole-2-yl) methanol (200 mg, 90% yield) as a colorless oil. MS (ESI): 139.1 [M+H] + . 1 H NMR (400MHz, CDCl3) δ5.99(s,1H),5.09(s,1H),4.65(d,J=1.6Hz,2H),4.16-4.12(m,2H),2.88(t,J=7.2Hz,2H),2.62–2.58(m,2H).

[0268] Step 5) 5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole-2-carbaldehyde

[0269] MnO2 (157 mg, 1.8 mmol) was added to a solution of (5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-2-yl)methanol (50 mg, 0.36 mmol) in chloroform (20 mL), and the mixture was heated to 60°C and stirred for 16 h. The mixture was filtered and the filtrate was concentrated to give 5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole-2-carboxaldehyde (30 mg, 58% yield) as a colorless oil. MS (ESI): 137.1 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.92 (s, 1H), 6.53 (s, 1H), 4.23 (t, J = 7.2Hz, 2H), 2.99-2.95 (m, 2H), 2.72-2.62 (m, 2H).

[0270] Step 6) 2-ethynyl-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole

[0271] To a solution of 5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole-2-carboxaldehyde (30 mg, 0.22 mmol) and potassium carbonate (61 mg, 0.44 mmol) in methanol (5 mL) was added dimethyl (1-diazo-2-oxopropyl)phosphonate (63 mg, 0.33 mmol), and the mixture was stirred at room temperature for 16 h. The solvent was removed by concentration, and the residue was diluted with ethyl acetate (30 mL). The mixture was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and separated by silica gel column chromatography to obtain 2-ethynyl-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole (22 mg, 61.8% yield) as a colorless oil. MS (ESI): 133.2 [M+H] + . 1 H NMR (400MHz, CDCl3) δ6.19(s,1H),4.28-4.17(m,2H),3.13(s,1H),2.96-2.87(m,2H),2.67-2.60(m,2H).

[0272] Step 7) (S)-3-(1-aminoethyl)-8-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-2-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one

[0273] Under nitrogen atmosphere, X-Phos (31.9 mg, 0.067 mmol), Pd2(dba)3 (30.6 mg, 0.033 mmol) and K3PO4 (85.3 mg, 0.40 mmol) were added to a solution (30 mL) of (S)-3-(1-aminoethyl)-8-chloro-2-phenylisoquinolin-1(2H)-one (40 mg, 0.13 mmol) and 2-ethynyl-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole (21.2 mg, 0.16 mmol) in acetonitrile, and the mixture was heated to 80°C and stirred for 5 h. The mixture was cooled to room temperature and concentrated under reduced pressure to remove the solvent. The residue was diluted with ethyl acetate (30 mL). The mixture was washed with saturated brine (20 mL), concentrated under reduced pressure to remove the solvent, and the residue was separated by flash silica gel column chromatography (DCM / MeOH=91 / 9) to obtain (S)-3-(1-aminoethyl)-8-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-2-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (60 mg, 90.9% yield) as a yellow oil. MS (ESI): 395.1 [M+H] + .

[0274] Step 8) (S)-2-amino-N-(1-(8-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-2-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-5,7-D2-3-carboxamide

[0275] At room temperature, DIEA (63.8 mg, 0.49 mmol) was added to an acetonitrile solution (10 mL) of (S)-3-(1-aminoethyl)-8-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-2-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (60 mg, 0.16 mmol) and 2,5-dioxopyrrolidin-1-yl 2-aminopyrazolo[1,5-a]pyrimidine-3-carboxylate-5,7-D2 (59 mg, 0.21 mmol), and the mixture was heated to 80°C and stirred for 16 h. The mixture was cooled to room temperature, concentrated under reduced pressure to remove the solvent, and diluted with dichloromethane (30 mL). The mixture was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure. The residue was purified by preparative HPLC (Gemini-C18 150x21.2mm, 5um ACN--H2O (0.1% FA) 45-50) to give (S)-2-amino-N-(1-(8-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-2-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-5,7-D2-3-carboxamide (5.3 mg, 5.8% yield) as a white solid. MS (ESI): 556.8 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.93 (dd, J=6.7, 1.6Hz, 1H), 8.56 (dd, J=4.5, 1.6Hz, 1H), 8.01 (d, J=6 .7Hz,1H),7.66(d,J=4.5Hz,2H),7.63-7.55(m,2H),7.53-7.46(m,3H),7.41-7.36(m,1H) ,7.02(dd,J=6.7,4.5Hz,1H),6.76(s,1H),6.44(s,2H),6.17(s,1H),4.56(t,J=6.8Hz,1H ), 4.09-4.02 (m, 2H), 2.82 (t, J = 7.3Hz, 2H), 2.54 (d, J = 7.7Hz, 1H), 1.35 (d, J = 6.8Hz, 3H).

[0276] Example 2 (S)-2-amino-5-methyl-N-(1-(8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0277]

[0278] Step 1) 1-Methyl-4-((trimethylsilyl)ethynyl)-1H-pyrazole

[0279] Trimethylethynylsilane (2.8 g, 28.8 mmol), CuI (366.24 mg, 1.923 mmol) and bis(triphenylphosphine)palladium(II)dichloride (1.3 g, 1.9 mmol) were added to a solution of 4-iodo-1-methylpyrazole (4 g, 19.2 mmol) in triethylamine (30 mL), heated to 90 °C and stirred for 16 h. After cooling to room temperature, the solvent was removed by concentration under reduced pressure. The residue was dissolved in DCM (300 mL), diluted with water (150 mL), and washed with saturated brine (150 mL x 2). The organic phase was dried over anhydrous sodium sulfate and concentrated. The residue was separated by silica gel column chromatography (DCM / MEOH=10 / 1) to give 1-methyl-4-((trimethylsilyl)ethynyl)-1H-pyrazole (2.5 g, 72.9% yield) as a brown oil. MS (ESI): 179.1 [M+H] + .

[0280] Step 2) 4-ethynyl-1-methyl-1H-pyrazole

[0281] To a solution of 1-methyl-4-((trimethylsilyl)ethynyl)-1H-pyrazole (2.3 g, 7.291 mmol) in THF (20 mL) was added tetrabutylammonium fluoride (14.6 mL, 14.6 mmol, 1 N in THF) at room temperature. The reaction was stirred at room temperature for 1 hour. The reaction was diluted with EtOAc (300 mL) and then washed with brine (150 mL×2). The separated organic layer was dried over Na2SO4 and concentrated in vacuo. The residue was purified by chromatography (PE:EA=10:1) to give 4-ethynyl-1-methyl-1H-pyrazole (0.9 g, 65% yield) as a clear oil. MS (ESI): 107.0 [M+H] + . 1 HNMR (400MHz, CDCl3) δ7.60(s,1H),7.52(s,1H),3.88(s,3H),3.01(s,1H).

[0282] Step 3) (S)-3-(1-aminoethyl)-8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one

[0283] To a mixture of (S)-3-(1-aminoethyl)-8-chloro-2-phenylisoquinolin-1(2H)-one (400 mg, 1.34 mmol) and 4-ethynyl-1-methyl-1H-pyrazole (213.1 mg, 2.00 mmol) in CH3CN (30 mL) at room temperature under nitrogen atmosphere were added X-Phos (319.1 mg, 0.67 mmol), Pd2(dba)3 (306.5 mg, 0.33 mmol) and K3PO4 (852.5 mg, 4.02 mmol), and the mixture was stirred at 80°C for 5 h. After cooling to room temperature, the mixture was concentrated in vacuo, and the residue was diluted with EtOAc (130 mL). The mixture was washed with brine (50 mL), dried over Na2SO4, and concentrated in vacuo. The residue was purified by flash chromatography (DCM / MeOH=91 / 9) to give (S)-3-(1-aminoethyl)-8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (460 mg, 83.93% yield) as a yellow oil. MS (ESI): 369.1 [M+H] + .

[0284] Step 4) (S)-2-amino-5-methyl-N-(1-(8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0285] To a solution of (S)-3-(1-aminoethyl)-8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (60 mg, 0.16 mmol) and 2-amino-5-methylpyrazolo[1,5-a]pyrimidine-3-carboxylic acid (31.31 mg, 0.16 mmol) in DCM (20 mL) was added DIEA (84.1 mg, 0.65 mmol), EDCI (46.8 mg, 0.24 mmol), HOAT (39.4 mg, 0.24 mmol). The mixture was stirred at 40° C. for 16 hours. The mixture was diluted with DCM (30 mL), then washed with brine (20 mL), dried over Na2SO4, and concentrated in vacuo. The residue was purified by preparative HPLC (Gemini-C18 150x21.2mm, 5um ACN--H2O (0.1% FA) 30-70) to give (S)-2-amino-5-methyl-N-(1-(8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (28.8 mg, 32.47% yield) as a white solid. MS (ESI): 545.9 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.78(d,J=6.9Hz,1H),8.17(d,J=6.6Hz,1H),8.01(s,1H),7.66-7.45(m,8H),7.40(d,J=7.6Hz,1H) ,6.91(d,J=6.9Hz,1H),6.73(s,1H),6.32(s,2H),4.526-4.49(m,1H),3.82(s,3H),2.57(s,3H),1.35(d,J=6.8Hz,3H).

[0286] Example 3 2-amino-N-((1S)-1-(1-oxo-8-(2-(5-oxopyrrolidin-3-yl)ethynyl)-2-phenylisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0287]

[0288] Step 1) 1-(2,4-dimethoxybenzyl)-5-oxopyrrolidine-3-carboxylic acid

[0289] A toluene (50 mL) solution of (2,4-dimethoxyphenyl)methylamine (2.0 g, 11.9 mmol) and 2-methylenediamine (1.55 g, 11.9 mmol) was heated to 100 ° C and stirred for 16 h. The mixture was diluted with water (20 mL) and then extracted with EtOAc (150 mL). The combined organic phases were washed with saturated brine (40 mL x 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM / MeOH=20 / 1) to obtain 1-(2,4-dimethoxybenzyl)-5-oxopyrrolidine-3-carboxylic acid (3.2 g, 91% yield) as a colorless oily liquid. MS (ESI): 280.1 [M+H] + .

[0290] Step 2) 1-[(2,4-dimethoxyphenyl)methyl]-4-(hydroxymethyl)pyrrolidin-2-one

[0291] At 0°C, BH3.THF (13 mL, 1 moL / L) was added to a solution of 1-(2,4-dimethoxybenzyl)-5-oxopyrrolidine-3-carboxylic acid (3.2 g, 11.4 mmol) in THF (50 mL), then the temperature was raised to room temperature and stirred for 16 h. The mixture was quenched with MeOH (20 mL) and concentrated under reduced pressure. The residue was dissolved in EtOAc (200 mL) and washed with saturated brine (40 mL x 2). The organic phase was dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM / MeOH=20 / 1) to give 1-[(2,4-dimethoxyphenyl)methyl]-4-(hydroxymethyl)pyrrolidin-2-one (2.0 g, 59% yield) as a colorless oily liquid. MS (ESI): 266.1 [M+H] + .

[0292] Step 3) 1-[(2,4-dimethoxyphenyl)methyl]-5-oxopyrrolidine-3-carbaldehyde

[0293] At 20°C, Dess-Martin periodinane (3.1 g, 6 mmol) was added to a solution of 1-[(2,4-dimethoxyphenyl)methyl]-4-(hydroxymethyl)pyrrolidin-2-one (1.6 g, 6.03 mmol) in DCM (50 mL), and the mixture was stirred at this temperature for 16 h. DCM (250 mL) was added to the mixture and filtered. The filtrate was washed with aqueous Na2SO3 solution (50 mL) and aqueous NaHCO3 solution (50 mL). The organic phase was dried over anhydrous Na2SO4 and concentrated under reduced pressure to give 1-[(2,4-dimethoxyphenyl)methyl]-5-oxopyrrolidine-3-carboxaldehyde (1.6 g, 80% yield) as a colorless oily liquid. MS (ESI): 264.1 [M+H]+ .

[0294] Step 4) 1-[(2,4-dimethoxyphenyl)methyl]-4-ethynylpyrrolidin-2-one

[0295] At 20°C, dimethyl (1-diazo-2-oxopropyl)phosphonate (1.38 g, 7.3 mmol) was added to a solution of 1-[(2,4-dimethoxyphenyl)methyl]-5-oxopyrrolidine-3-carboxaldehyde (1.6 g, 6.1 mmol) and K2CO3 (2.5 g, 18 mmol) in MeOH (100 mL), and the mixture was stirred for 4 h. The mixture was diluted with EtOAc (250 mL) and washed with saturated brine (50 mL x 2). The organic phase was dried over anhydrous Na2SO4 and concentrated under reduced pressure to give 1-[(2,4-dimethoxyphenyl)methyl]-4-ethynylpyrrolidine-2-one (1.1 g, 62% yield) as a colorless oily liquid. MS (ESI): 260.1 [M+H] + .

[0296] Step 5) 3-((1S)-1-aminoethyl)-8-(2-(1-((2,4-dimethoxyphenyl)methyl)-5-oxopyrrolidin-3-yl)ethynyl)-2-phenylisoquinolin-1-one

[0297] Under nitrogen atmosphere, K3PO4 (341 mg, 1.6 mmol), x-Phos (2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl, 102 mg, 0.21 mmol) and Pd2(dba)3 (98 mg, 0.1 mmol) were added to a solution of 1-[(2,4-dimethoxyphenyl)methyl]-4-ethynylpyrrolidin-2-one (208 mg, 0.8 mmol) and (S)-3-(1-aminoethyl)-8-chloro-2-phenylisoquinolin-1(2H)-one (160 mg, 0.53 mmol) in acetonitrile (50 mL). The mixture was heated to 80°C and stirred for 16 h. After the reaction was complete, the mixture was cooled to room temperature and concentrated under reduced pressure. The residue was diluted with EtOAc (200 mL), washed with saturated brine (100 mL), and the organic phase was dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM / MeOH=20 / 1) to give 3-((1S)-1-aminoethyl)-8-(2-(1-((2,4-dimethoxyphenyl)methyl)-5-oxopyrrolidin-3-yl)ethynyl)-2-phenylisoquinolin-1-one (280 mg, 89% yield) as a brown solid. MS (ESI): 522.2 [M+H] + .

[0298] Step 6) 2-amino-N-((1S)-1-(8-(2-(1-((2,4-dimethoxyphenyl)methyl)-5-oxopyrrolidin-3-yl)ethynyl)-1-oxo-2-phenylisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0299] To a solution of 2-aminopyrazolo[1,5-a]pyrimidine-3-carboxylic acid (95 mg, 0.53 mmol), HOAt (109 mg, 0.8 mmol), 3-((1S)-1-aminoethyl)-8-(2-(1-((2,4-dimethoxyphenyl)methyl)-5-oxopyrrolidin-3-yl)ethynyl)-2-phenylisoquinolin-1-one (280 mg, 0.53 mmol) and EDCI (154 mg, 0.80 mmol) in DCM (10 mL) was added DIEA (208 mg, 1.6 mmol) at room temperature. The mixture was heated to 40°C and stirred for 16 h. DCM (60 mL) was added to the mixture to dilute it and then washed with saturated brine (50 mL). The organic phase was dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM / MeOH=20 / 1) to give 2-amino-N-((1S)-1-(8-(2-(1-((2,4-dimethoxyphenyl)methyl)-5-oxopyrrolidin-3-yl)ethynyl)-1-oxo-2-phenylisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (320 mg, 78% yield) as a brown solid. MS (ESI): 682.3 [M+H] + .

[0300] Step 7) 2-amino-N-((1S)-1-(1-oxo-8-(2-(5-oxopyrrolidin-3-yl)ethynyl)-2-phenylisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0301] 2-Amino-N-((1S)-1-(8-(2-(1-((2,4-dimethoxyphenyl)methyl)-5-oxopyrrolidin-3-yl)ethynyl)-1-oxo-2-phenylisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (150 mg, 0.22 mmol) was added to 6N HCl (10 mL), the temperature was raised to 80°C and stirred for 1 h. The residue was concentrated under reduced pressure and purified by preparative HPLC (CAN-H2O, 0.1% FA, gradient 20% to 50%) to give 2-amino-N-((1S)-1-(1-oxo-8-(2-(5-oxopyrrolidin-3-yl)ethynyl)-2-phenylisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (3.7 mg, 3.1% yield) as a white solid. MS (ESI): 532.2 [M+H] + . 1 H NMR(400MHz,CD3OD)δ8.72(d,J=6.8Hz,1H),8.52(s,1H),8.43(t,J=6.4Hz,1H),8.26 (t,J=7.2Hz,1H),8.12(d,J=7.2Hz,1H),8.04(d,J=8.0Hz,1H),8.00(s,1H),7.93(d, J=8.0Hz,1H),7.80-7.71(m,3H),7.02-6.98(m,1H),6.72(d,J=8.8Hz,1H),5.09-5.0 5(m,1H),3.66-3.52(m,3H),2.56-2.48(m,1H),2.35-2.30(m,1H),1.65-1.57(m,3H).

[0302] Example 4 2-amino-N-((1S)-1-(8-(2-(1-methyl-5-oxopyrrolidin-3-yl)ethynyl)-1-oxo-2-phenylisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0303]

[0304] Step 1) 4-(Hydroxymethyl)-1-methylpyrrolidin-2-one

[0305] At 0°C, NaBH4 (481 mg, 12.7 mmol) was added to a MeOH (50 mL) solution of 1-methyl-5-oxopyrrolidine-3-carboxylic acid methyl ester (1 g, 6.3 mmol). The mixture was warmed to room temperature, stirred for 16 h, and then concentrated under reduced pressure. The residue was dissolved with MeOH / DCM=1 / 20 (200 mL) and filtered. The filtrate was dried over anhydrous Na2SO4, concentrated under reduced pressure, and the residue was separated by column chromatography (MeOH / DCM=1 / 20) to obtain 4-(hydroxymethyl)-1-methylpyrrolidin-2-one (900 mg, 98.57% yield) as a colorless oily liquid. MS (ESI): 130.1 [M+H] + . 1 H NMR (400MHz, CDCl3) δ3.62-3.57(m,2H),3.50(dd,J=10.0,8.0Hz,1H),3.26(dd,J=10.0,5. 0Hz, 1H), 2.84 (s, 3H), 2.63-2.55 (m, 1H), 2.55-2.49 (m, 1H), 2.22 (dd, J = 16.4, 5.2Hz, 1H).

[0306] Step 2) 1-methyl-5-oxopyrrolidine-3-carbaldehyde

[0307] At 20°C, Dess-Martin periodinane (2.7 g, 6.5 mmol) was added to a solution of 4-(hydroxymethyl)-1-methylpyrrolidin-2-one (420 mg, 3.2 mmol) in DCM (50 mL), and the mixture was stirred at this temperature for 16 h. DCM (100 mL) was added to the mixture and filtered. The filtrate was washed with an aqueous Na2SO3 solution (50 mL) and an aqueous NaHCO3 solution (50 mL). The organic phase was dried over anhydrous Na2SO4, concentrated under reduced pressure, and the residue was separated by column chromatography (MeOH / DCM=1 / 20) to give 1-methyl-5-oxopyrrolidine-3-carbaldehyde (280 mg, 67.72% yield) as a colorless oily liquid. MS (ESI): 128.1 [M+H] + .

[0308] Step 3) 4-ethynyl-1-methylpyrrolidin-2-one

[0309] At 20°C, dimethyl (1-diazo-2-oxopropyl)phosphonate (718 mg, 3.78 mmol) was added to a solution of 1-methyl-5-oxopyrrolidine-3-carboxaldehyde (400 mg, 3.1 mmol) and K2CO3 (1.3 g, 9.4 mmol) in MeOH (30 mL), and the mixture was stirred for 4 h. The mixture was diluted with H2O (10 mL) and washed with saturated brine (40 mL x 2). The organic phase was dried over anhydrous Na2SO4, concentrated under reduced pressure, and the residue was separated by column chromatography (MeOH / DCM=1 / 30) to obtain 4-ethynyl-1-methylpyrrolidin-2-one (180 mg, 37.17% yield) as a colorless oily liquid. MS (ESI): 124.1 [M+H] + . 1 HNMR (400MHz, DMSO-d6) δ3.70-3.55(m,2H),3.27(dd,J=9.1,6.3Hz,1H),2.71(s,3H),2.56(dd,J=16.3,8.6Hz,1H),2.22(dd,J=16.3,7.1Hz,1H).

[0310] Step 4) 3-((1S)-1-aminoethyl)-8-(2-(1-methyl-5-oxopyrrolidin-3-yl)ethynyl)-2-phenylisoquinolin-1-one

[0311] Under nitrogen atmosphere, K3PO4 (128 mg, 0.6 mmol), x-Phos (38 mg, 0.08 mmol) and Pd2(dba)3 (37 mg, 0.04 mmol) were added to a solution of 4-ethynyl-1-methylpyrrolidin-2-one (37.1 mg, 0.3 mmol) and (S)-3-(1-aminoethyl)-8-chloro-2-phenylisoquinolin-1(2H)-one (60 mg, 0.2 mmol) in acetonitrile (20 mL). The mixture was heated to 80°C and stirred for 4 h. After the reaction was complete, the mixture was cooled to room temperature and concentrated under reduced pressure. The residue was diluted with EtOAc (60 mL) and washed with saturated brine (30 mL x 2). The organic phase was dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM / MeOH=20 / 1) to give 3-((1S)-1-aminoethyl)-8-(2-(1-methyl-5-oxopyrrolidin-3-yl)ethynyl)-2-phenylisoquinolin-1-one (30 mg, 35.0% yield) as a yellow solid. MS (ESI): 473.0 [M+H] + .

[0312] Step 5) 2-amino-N-((1S)-1-(8-(2-(1-methyl-5-oxopyrrolidin-3-yl)ethynyl)-1-oxo-2-phenylisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0313] To a solution of 2-aminopyrazolo[1,5-a]pyrimidine-3-carboxylic acid (15 mg, 0.08 mmol), HOAt (16 mg, 0.12 mmol), 3-((1S)-1-aminoethyl)-8-(2-(1-methyl-5-oxopyrrolidin-3-yl)ethynyl)-2-phenylisoquinolin-1-one (30 mg, 0.08 mmol) and EDCI (22 mg, 0.12 mmol) in DCM (10 mL) was added DIEA (30 mg, 0.23 mmol) at room temperature. The mixture was heated to 40°C and stirred for 16 h. DCM (60 mL) was added to the mixture to dilute it and then washed with saturated brine (40 mL). The organic phase was dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was separated by preparative HPLC (CAN-H2O 0.1FA, gradient 30% to 60%) to give 2-amino-N-((1S)-1-(8-(2-(1-methyl-5-oxopyrrolidin-3-yl)ethynyl)-1-oxo-2-phenylisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (18.0 mg, 42.42% yield) as a white solid. MS (ESI): 546.2 [M+H] + . 1 H NMR (400MHz, CDCl3) δ8.47(d,J=6.8Hz,1H),8.42(d,J=4.5Hz,1H),7.94-7.89(m,1H),7.53(d,J= 1.7Hz,1H),7.51(d,J=7.5Hz,1H),7.47(d,J=1.3Hz,3H),7.42(s,1H),7.35(d,J=8.0Hz,1H),6.8 2(dd,J=6.6,4.5Hz,1H),6.62(s,1H),4.84-4.78(m,1H),3.63(dd,J=12.8,5.0Hz,1H),3.57-3.5 1(m,1H),3.51-3.43(m,1H),2.84(s,3H),2.78-2.71(m,1H),2.63(s,1H),1.40(d,J=6.8Hz,3H).

[0314] Example 5 (S)-2-amino-N-(1-(8-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-ylethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0315]

[0316] Step 1) 5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole-3-carbaldehyde

[0317] At -78 ° C, to a solution of 3-bromo-4H,5H,6H-pyrrolo[1,2-b]pyrazole (200 mg, 1.07 mmol) in tetrahydrofuran (10 mL) was slowly added n-butyl lithium (2.4 M in n-hexane, 0.49 mL, 1.18 mmol), and the mixture was stirred at -70 ° C for 1 hour, and then a solution of N,N-dimethylformamide (156 mg, 2.1 mmol) in tetrahydrofuran (1 mL) was added, and the mixture was stirred at 0 ° C for 2 hours. The mixture was quenched with aqueous ammonium chloride (10 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layer was washed with saturated brine (10 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel chromatography (EA / PE=1 / 3) to give 5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole-3-carbaldehyde (100 mg, 65% yield) as a white solid. MS (ESI): 137.1 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.81(s,1H),7.95(s,1H),4.25-4.16(m,2H),3.18-3.10(m,2H),2.75-2.63(m,2H).

[0318] Step 2) 3-ethynyl-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole

[0319] To a solution of 5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole-3-carboxaldehyde (100 mg, 0.73 mmol) and potassium carbonate (203 mg, 1.47 mmol) in methanol (10 mL) was added dimethyl (1-diazo-2-oxopropyl)phosphonate (212 mg, 1.1 mmol), and the mixture was stirred at 25°C for 16 hours, concentrated under reduced pressure, and the residue was diluted with ethyl acetate (40 mL). The organic layer was washed with saturated brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 3-ethynyl-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole (80 mg, 70% yield) as a colorless oil. MS (ESI): 133.1 [M+H] + . 1 H NMR (400MHz, CDCl3) δ7.61(s,1H),4.16-4.11(m,2H),3.03(s,1H),3.00-2.92(m,2H),2.67-2.60(m,2H).

[0320] Step 3) (S)-3-(1-aminoethyl)-8-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one

[0321] Under nitrogen atmosphere, X-Phos (48 mg, 0.10 mmol), Pd2(dba)3 (46 mg, 0.05 mmol) and potassium phosphate (128 mmol, 0.60 mmol) were added to a solution of (S)-3-(1-aminoethyl)-8-chloro-2-phenylisoquinolin-1(2H)-one (60 mg, 0.20 mmol) and 3-ethynyl-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole (40 mg, 0.30 mmol) in acetonitrile (30 mL). The mixture was heated at 80°C for 5 hours, cooled to room temperature, and then concentrated under reduced pressure. The residue was diluted with ethyl acetate (30 mL), washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel chromatography (DCM / MeOH=91 / 9) to give (S)-3-(1-aminoethyl)-8-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (35 mg, 39.8% yield) as a yellow oil. MS (ESI): 394.9 [M+H] + .

[0322] Step 4) (S)-2-amino-N-(1-(8-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-ylethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0323] To a solution of (S)-3-(1-aminoethyl)-8-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-pyrazol-3-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (37 mg, 0.09 mmol) and 2-amino-5-methylpyrazolo[1,5-a]pyrimidine-3-carboxylic acid (18 mg, 0.10 mmol) in dichloromethane (20 mL) was added N,N'-diisopropylethylamine (36 mg, 0.28 mmol), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (27 mg, 0.14 mmol) and 1-hydroxy-7-azabenzotriazole (19 mg, 0.14 mmol) at room temperature. The mixture was stirred at 40°C for 16 hours, then diluted with dichloromethane (30 mL), washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by high performance preparative liquid phase (Gemini-C18 150x21.2 mm, 5um ACN-H2O (0.1% FA) 30-70) to give (S)-2-amino-N-(1-(8-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinoline-3-ethyl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (9.3 mg, 17.70% yield) as a white solid. MS (ESI): 554.8 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.93 (dd, J=6.7, 1.6Hz, 1H), 8.55 (dd, J=4.5, 1.6Hz, 1H) ,8.00(d,J=6.7Hz,1H),7.66-7.53(m,5H),7.52-7.46(m,3H),7.42-7.34(m,1H), 7.02(dd,J=6.7,4.5Hz,1H),6.73(s,1H),6.44(s,2H),4.55(q,J=6.7Hz,1H),4. 07(t,J=7.2Hz,2H),2.93-2.80(m,2H),2.58-2.53(m,1H),1.35(d,J=6.8Hz,3H).

[0324] Example 6 2-amino-N-((1S)-1-(1-oxo-8-((5-oxotetrahydrofuran-3-yl)ethynyl)-2-phenyl-1,2-dihydroisoquinolin-3-alkyl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0325]

[0326] Step 1) 4-(1-bromovinyl)dihydrofuran-2(3H)-one

[0327] At 0°C, liquid bromine (784 mg, 4.9 mmol) was added to a solution of 4-vinyloxacyclo-2-one (500 mg, 4.46 mmol) in dichloromethane (40 mL). After the mixture was stirred at 0°C for 2 hours, the mixture was concentrated under reduced pressure and the residue was dissolved in acetonitrile (20 mL). 1,8-diazabicycloundec-7-ene (1.35 g, 8.9 mmol) was added to the mixture at 0°C and stirred at 25°C for 16 hours. The mixture was diluted with ethyl acetate (50 mL) and washed with saturated brine (30 mL x 2). The separated organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (EA / PE=1 / 4) to give 4-(1-bromovinyl)dihydrofuran-2(3H)-one (600 mg, 31.7% yield) as a brown oil. MS(ESI):191.0,193.0[M+H] + . 1 H NMR (400MHz, CDCl3) δ5.80 (dd, J=2.4, 0.8Hz, 1H), 5.58 (d, J=2.4Hz, 1H), 4.45-4.39 (m, 1H), 4.25-4.20 (m, 1H), 3.55-3.46 (m, 1H), 2.71-2.66 (m, 2H).

[0328] Step 2) 4-Ethyldihydrofuran-2(3H)-one

[0329] At -70°C, n-butyllithium (2.4N n-hexane solution, 5.9 mL, 14.3 mmol) was slowly added dropwise to a solution of diisopropylamine (1.38 g, 13.6 mmol) in tetrahydrofuran (30 mL), and stirred at -30°C for 1 hour. The mixture was then cooled to -70°C, a solution of 4-(1-bromovinyl)dihydrofuran-2(3H)-one (650 mg, 3.4 mmol) in tetrahydrofuran (3 mL) was added, and stirred at -10°C for 2 hours. The mixture was quenched with aqueous ammonium chloride solution (30 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layer was washed with saturated brine (50 mL), and the separated organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (EA / PE=1 / 4) to give 4-ethynyldihydrofuran-2(3H)-one (250 mg, 43.4% yield) as a colorless oil. 1 H NMR (400MHz, CDCl3) δ4.51 (dd, J=8.8, 7.6Hz, 1H), 4.22 (dd, J=8.8, 7.6Hz, 1H), 3.45-3.33 (m, 1H), 2.81 (dd, J = 17.4, 8.8 Hz, 1H), 2.63 ( dd, J = 17.4, 8.8 Hz, 1H), 2.25 ( d, J = 2.4 Hz, 1H).

[0330] Step 3) 3-((S)-1-aminoethyl)-8-((5-oxotetrahydrofuran-3-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one

[0331] Under nitrogen atmosphere, X-Phos (40 mg, 0.083 mmol), Pd2(dba)3 (38 mg, 0.042 mmol) and potassium phosphate (107 mg, 0.50 mmol) were added to a solution of (S)-3-(1-aminoethyl)-8-chloro-2-phenylisoquinolin-1(2H)-one (50 mg, 0.17 mmol) and 4-ethynyldihydrofuran-2(3H)-one (37 mg, 0.33 mmol) in acetonitrile (30 mL), and the mixture was heated at 80°C for 5 hours. After cooling to room temperature, the mixture was concentrated under reduced pressure, and the residue was diluted with ethyl acetate (30 mL). The mixture was washed with saturated brine (20 mL), and the separated organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (DCM / MeOH=91 / 9) to give 3-((S)-1-aminoethyl)-8-((5-oxotetrahydrofuran-3-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (48 mg, yield 69.30%) as a yellow oil. MS (ESI): 373.0 [M+H] + .

[0332] Step 4) 2-amino-N-((1S)-1-(1-oxo-8-((5-oxotetrahydrofuran-3-yl)ethynyl)-2-phenyl-1,2-dihydroisoquinolin-3-alkyl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0333] To a mixture of 3-((S)-1-aminoethyl)-8-((5-oxotetrahydrofuran-3-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (48 mg, 0.13 mmol) and 2,5-dioxopyrrolidin-1-yl 2-aminopyrazolo[1.5-a]pyrimidine-3-carboxylate (35.5 mg, 0.13 mmol) in acetonitrile (10 mL) was added N,N'-diisopropylethylamine (50 mg, 0.39 mmol), and the mixture was stirred at 80°C for 16 hours. After cooling to room temperature, the mixture was concentrated under reduced pressure, the residue was diluted with dichloromethane (30 mL), washed with saturated brine (20 mL), the separated organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue (Gemini-C18 150x21.2mm.2mm.5um ACN-H2O (0.1% FA) 40-55) was purified to give 2-amino-N-((1S)-1-(1-oxo-8-((5-oxotetrahydrofuran-3-yl)ethynyl)-2-phenyl-1,2-dihydroisoquinoline-3-ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (14.8 mg, 21.1% yield) as a white solid. MS (ESI): 532.8 [M+H] + . 1 HNMR(400MHz,DMSO-d6)δ8.93(dd,J=6.7,1.6Hz,1H),8.55(dd,J=4.5,1.6Hz,1H),7.99( d,J=6.7Hz,1H),7.67-7.42(m,7H),7.37(dd,J=5.7,3.5Hz,1H),7.01(dd,J=6.7,4.5Hz,1 H),6.74(s,1H),6.43(s,2H),4.59-4.47(m,2H),4.20(dd,J=8.3,6.9Hz,1H),3.77-3.68( m, 1H), 2.86 (dd, J = 17.0, 8.6 Hz, 1H), 2.61 ( dd, J = 17.0, 7.8 Hz, 1H), 1.34 ( d, J = 6.8 Hz, 3H).

[0334] Example 7 (S)-2-amino-N-(1-(8-((3-hydroxyoxetan-3-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-alkyl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0335]

[0336] Step 1) 3-ethynyloxetane-3-ol

[0337] At -78 ° C, under nitrogen protection, ethynylmagnesium bromide (25mL, 12.50mmol, 0.5mmol in THF solution) was slowly added to a solution of 3-oxetanone (750mg, 10.41mmol) in tetrahydrofuran (20mL), and the mixture was stirred at room temperature for 2 hours. The reaction was quenched with aqueous ammonium chloride (20mL) and extracted with ethyl acetate (50mL x 3). The combined organic layer was washed with saturated brine (50mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by flash silica gel chromatography (DCM / MeOH=30 / 1) to give 3-ethynyloxetane-3-ol (485mg, 42.75% yield) as a yellow oil. 1 H NMR (400MHz, CDCl3) δ4.83 (dd, J = 6.7, 0.9 Hz, 2H), 4.68 (dd, J = 6.6, 0.9 Hz, 2H), 3.52 (s, 1H), 2.71 (s, 1H).

[0338] Step 2) 3-((S)-1-aminoethyl)-8-((3-hydroxyoxetan-3-yl)ethynyl)-2-phenylnaphthalen-1(2H)-one

[0339] To a solution of (S)-3-(1-aminoethyl)-8-chloro-2-phenylisoquinolin-1(2H)-one (60 mg, 0.20 mmol) and 3-ethynyloxyalkane-3-ol (29 mg, 0.30 mmol) in acetonitrile (10 mL) were added X-Phos (48 mg, 0.10 mmol), Pd2(dba)3 (37 mg, 0.04 mmol) and potassium phosphate (128 mg, 0.60 mmol), and the mixture was heated at 80°C for 5 hours. After cooling to room temperature, the mixture was concentrated under reduced pressure, and the residue was diluted with ethyl acetate (60 mL). The mixture was washed with saturated brine (40 mL), and the separated organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (DCM / MeOH=91 / 9) to give 3-((S)-1-aminoethyl)-8-((3-hydroxyoxetan-3-yl)ethynyl)-2-phenylnaphthalen-1(2H)-one (50 mg, yield 62.20%) as a yellow solid. MS (ESI): 361.0 [M+H] + .

[0340] Step 3) (S)-2-amino-N-(1-(8-((3-hydroxyoxetan-3-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-alkyl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0341] To a solution of 3-((S)-1-aminoethyl)-8-((3-hydroxyoxetan-3-yl)ethynyl)-2-phenylnaphthalene-1(2H)-one (25 mg, 0.07 mmol) and 2-aminopyrazolo[1.5-a]pyrimidine-3-carboxylic acid (12 mg, 0.07 mmol) in dichloromethane (20 mL) were added N,N'-diisopropylethylamine (27 mg, 0.21 mmol), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (20 mg, 0.10 mmol) and 1-hydroxy-7-azabenzotriazole (14 mg, 0.10 mmol). The mixture was stirred at 40°C for 16 hours, then the mixture was diluted with dichloromethane (60 mL), washed with saturated brine (40 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by high performance preparative liquid chromatography (-Gemini-C18 150x21.2mm, 5um: ACN-H2O (0.1% FA), 40-60) to give (S)-2-amino-N-(1-(8-((3-hydroxyoxetane-3-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinoline-3-ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (4.1 mg, 11.24%) as a yellow solid. MS (ESI): 520.9 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.93 (dd, J=6.7, 1.6Hz, 1H), 8.55 (dd, J=4.5, 1.6Hz, 1H),8.00(d,J=6.7Hz,1H),7.70-7.63(m,2H),7.60-7.54(m,2H),7.53-7.45( m,3H),7.41-7.35(m,1H),7.02(dd,J=6.7,4.5Hz,1H),6.75(s,1H),6.58-6.3 5(m,3H),4.73(d,J=6.3Hz,2H),4.55(t,J=6.1Hz,3H),1.35(d,J=6.8Hz,3H).

[0342] Example 8 2-amino-N-((1S)-1-(8-(2-(1,2-dimethyl-5-oxopyrazol-3-yl)ethynyl)-1-oxo-2-phenylisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0343]

[0344] Step 1) 1,2-dimethyl-5-oxopyrazole-3-carboxylic acid methyl ester

[0345] At 0°C, NaH (184 mg, 7.7 mmol) was added to a solution of 2-methyl-5-oxo-1H-pyrazole-3-carboxylic acid methyl ester (1 g, 6.4 mmol) in DMF (20 mL), and the mixture was stirred for 0.5 h. MeI (1.3 g, 9.6 mmol) was then added, followed by stirring for 4 h. The mixture was quenched with water, diluted with NH4Cl solution (50 mL), and extracted with EtOAc (50 mL x 3). The combined organic layers were washed with brine (100 mL x 2), dried over Na2SO4, and concentrated in vacuo. The residue was purified by silica gel chromatography (EA: PE = 1: 2) to give 1,2-dimethyl-5-oxo-pyrazole-3-carboxylic acid methyl ester (1.05 g, 91.5% yield) as a brown solid. MS (ESI): 171.1 [M+H] + .

[0346] Step 2) 5-(Hydroxymethyl)-1,2-dimethylpyrazol-3-one

[0347] At 0°C, LiAlH4 (446 mg, 11.7 mmol) was added to a solution of 1,2-dimethyl-5-oxopyrazole-3-carboxylic acid methyl ester (1 g, 5.9 mmol) in THF (50 mL), and the mixture was stirred for 2 h. The mixture was quenched by adding 10% NaOH (0.5 mL) and water (1 mL), and filtered. The filter cake was washed with EtOAc (40 mL), and the filtrate was dried over Na2SO4 and concentrated in vacuo to give crude 5-(hydroxymethyl)-1,2-dimethylpyrazol-3-one (740 mg, yield 88%) as a colorless oil. MS (ESI): 143.1 [M+H] + .

[0348] Step 3) 1,2-dimethyl-5-oxopyrazole-3-carboxaldehyde

[0349] At 20°C, Dess-Martin reagent (4.4 g, 10.4 mmol) was added to a solution of 5-(hydroxymethyl)-1,2-dimethylpyrazol-3-one (740 mg, 5.2 mmol) in DCM (100 mL), and the mixture was stirred for 16 hours. The mixture was diluted with DCM (100 mL) and filtered. The filtrate was washed with water (50 mL) and brine (50 mL), dried over Na2SO4, and concentrated in vacuo. The residue was purified by silica gel chromatography (EA / PE=1 / 1) to give 1,2-dimethyl-5-oxopyrazole-3-carboxaldehyde (710 mg, 87.5% yield) as a yellow oil. MS (ESI): 141.1 [M+H] + .

[0350] Step 4) 5-ethynyl-1,2-dimethylpyrazol-3-one

[0351] Dimethyl (1-diazo-2-oxopropyl) phosphonate (1.7 g, 6.1 mmol) was added to a solution of 1,2-dimethyl-5-oxopyrazole-3-carboxaldehyde (710 mg, 5.1 mmol) and K2CO3 (2.1 g, 15 mmol) in MeOH (50 mL), and the mixture was stirred at room temperature for 16 hours. After the mixture was concentrated in vacuo, it was diluted with EtOAc (100 mL); then washed with brine (100 mL), dried over Na2SO4, and concentrated in vacuo. The residue was purified by silica gel chromatography (EA:PE=1:1) to give 5-ethynyl-1,2-dimethylpyrazol-3-one (540 mg, 70.4% yield) as a colorless oil. MS (ESI): 137.1 [M+H] + .

[0352] Step 5) 3-((1S)-1-aminoethyl)-8-(2-(1,2-dimethyl-5-oxopyrazol-3-yl)ethynyl)-2-phenylisoquinolin-1-one

[0353] A solution of 5-ethynyl-1,2-dimethylpyrazol-3-one (9.6 mg, 0.07 mmol), 3-[(1S)-1-aminoethyl]-8-chloro-2-phenylisoquinolin-1-one (13.1 mg, 0.044 mmol), K3PO4 (28 mg, 0.13 mmol), X-phos (8.4 mg, 0.02 mmol) and Pd2(dba)3 (8.1 mg, 0.01 mmol) in CH3CN (10 mL) was heated to 80°C and stirred for 16 hours. The mixture was cooled to room temperature and concentrated in vacuo, and the residue was diluted with EtOAc (50 mL). The mixture was washed with brine (20 mL), dried over Na2SO4, and concentrated in vacuo. The residue was purified by silica gel chromatography (MeOH / DCM=1 / 20) to give 3-((1S)-1-aminoethyl)-8-(2-(1,2-dimethyl-5-oxopyrazol-3-yl)ethynyl)-2-phenylisoquinolin-1-one (120 mg, 95% yield) as a brown solid. MS (ESI): 399.2 [M+H] + .

[0354] Step 6) 2-amino-N-((1S)-1-(8-(2-(1,2-dimethyl-5-oxopyrazol-3-yl)ethynyl)-1-oxo-2-phenylisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0355] 3-((1S)-1-aminoethyl)-8-(2-(1,2-dimethyl-5-oxopyrazol-3-yl)ethynyl)-2-phenylisoquinolin-1-one (120 mg, 0.30 mmol) was added to a solution of 2-aminopyrazolo[1,5-a]pyrimidine-3-carboxylic acid (54 mg, 0.3 mmol) and HOAT (61.5 mg, 0.45 mmol), EDCI (86.6 mg, 0.45 mmol), and DIEA (117 mg, 0.9 mmol) in DCM (20 mL), and the mixture was heated to 40°C and stirred for 16 h. After cooling to room temperature, the mixture was diluted with DCM (60 mL). The mixture was washed with brine (50 mL), dried over Na2SO4, and concentrated in vacuo. The residue was purified by preparative HPLC (ACN-H2O 0.1% FA, gradient 30% to 50%) to give 2-amino-N-((1S)-1-(8-(2-(1,2-dimethyl)-5-oxopyrazol-3-yl)ethynyl)-1-oxo-2-phenylisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (56 mg, 32.6% yield) as a white solid. MS (ESI): 559.2 [M+H] + . 1H NMR (400MHz, CDCl3) δ8.49(dd,J=6.8,1.6Hz,1H),8.44(dd,J=4.4,1.6Hz,1H),7.97(d,J=6.8Hz,1H),7.66(dd,J=7.6,1.2Hz,1H),7.60-7.56(m,1H),7 .55-7.48(m,4H),7.38-7.33(m,1H),6.86-6.82(m,1H),6.67(s,1H),5.88( s,1H),4.86-4.81(m,1H),3.93(s,3H),3.91(s,3H),1.42(d,J=6.8Hz,3H).

[0356] Example 9 (S)-2-amino-N-(1-(8-((6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-2-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0357]

[0358] Step 1) 2-((triisopropylsilyl)ethynyl)-6,7-dihydro-5H-pyrrolo[1,2-a]imidazole

[0359] K2CO3 (443 mg, 3.2 mmol) and X-phos (76 mg, 0.16 mmol) were added to a DMF (10 mL) solution of 2-bromo-5H, 6H, 7H-pyrrolo[1,2-a]imidazole (300 mg, 1.6 mmol), ethynyltriisopropylsilane (439 mg, 2.4 mmol) and palladium acetate (36 mg, 0.16 mmol), and the mixture was heated to 80°C and stirred for 16 h. After cooling to room temperature, the mixture was diluted with EtOAc (50 mL) and washed with brine (30 mL×2). The organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was separated and purified by column chromatography (DCM / MeOH=20 / 1) to give the product (200 mg, 41% yield) as a yellow solid. MS (ESI): 289.2 [M+H] + .

[0360] Step 2) 2-((triisopropylsilyl)ethynyl)-6,7-dihydro-5H-pyrrolo[1,2-a]imidazole

[0361] Tetrabutylammonium fluoride (0.8 mL, 0.8 mmol, 1 M in THF) was added to a solution of 2-(2-(triisopropylsilyl)ethynyl)-5H,6H,7H-pyrrolo[1,2-a]imidazole (150 mg, 0.52 mmol) in THF (5 mL), and the mixture was heated to 25°C and stirred for 2 hours. The mixture was diluted with EtOAc (50 mL) and washed with brine (30 mL). The organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was separated by column chromatography (DCM / MeOH=15 / 1) to give 2-ethynyl-6,7-dihydro-5H-pyrrolo[1,2-a]imidazole (60 mg, yield 78%) as a white solid. MS (ESI): 133.0 [M+H] + .

[0362] Step 3) (S)-3-(1-aminoethyl)-8-((6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-2-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one

[0363] Under nitrogen atmosphere, to a solution of (S)-3-(1-aminoethyl)-8-chloro-2-phenylisoquinolin-1(2H)-one (60 mg, 0.20 mmol) and 2-ethynyl-6,7-dihydro-5H-pyrrolo[1,2-a]imidazole (34.5 mg, 0.26 mmol) in MeCN (50 mL) were added X-Phos (38 mg, 0.08 mmol), Pd2(dba)3 (37 mg, 0.04 mmol) and K3PO4 (128 mg, 0.60 mmol). The mixture was heated to 80°C and stirred for 5 h. After cooling to room temperature, the mixture was concentrated in vacuo, and the residue was diluted with EtOAc (130 mL) and washed with brine (50 mL×2). The organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=91 / 9) to give (S)-3-(1-aminoethyl)-8-((6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-2-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (50 mg, 57% yield) as a yellow oil. MS (ESI): 394.9 [M+H] + .

[0364] Step 4) (S)-2-amino-N-(1-(8-((6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-2-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0365] To a solution of (S)-3-(1-aminoethyl)-8-((6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-2-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (50 mg, 0.13 mmol) and 2-aminopyrazolo[1,5-a]pyrimidine-3-carboxylic acid (25 mg, 0.14 mmol) in DCM (20 mL) were added DIEA (65 mg, 0.51 mmol), EDCI (36 mg, 0.19 mmol) and HOAT (26 mg, 0.19 mmol). The mixture was warmed to 40 °C and stirred for 16 h. After cooling to room temperature, the mixture was diluted with DCM (100 mL) and washed with brine (50 mL x 2). The organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by preparative HPLC (-Xbridge-C18 150x 19mm, 5um, ACN-H2O (0.05% NH4OH), 15%-40%) to give (S)-2-amino-N-(1-(8-((6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-2-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (13.4 mg, 19.01% yield) as a white solid. MS (ESI): 554.8 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ8.94(dd,J=6.7,1.6Hz,1H),8.56(dd,J=4.5,1.6Hz,1H),8 .02(d,J=6.6Hz,1H),7.89(s,1H),7.79-7.65(m,3H),7.61-7.46(m,4H),7.43-7.38 (m,1H),7.02(dd,J=6.7,4.5Hz,1H),6.81(s,1H),6.43(s,2H),4.60-4.52(m,1H), 4.17-4.09(m,2H),3.04(t,J=7.6Hz,2H),2.66-2.55(m,2H),1.36(d,J=6.8Hz,3H).

[0366] Example 10 2-amino-5-(fluoromethyl-d)-N-((S)-1-(8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0367]

[0368] Step 1) 2-amino-5-methylpyrazolo[1,5-a]pyrimidine-3-carboxylic acid ethyl ester

[0369] 4,4-dimethoxybutan-2-one (3.1 g, 23.5 mmol) was added to a solution of ethyl 3,5-diamino-1H-pyrazole-4-carboxylate (2 g, 11.7 mmol) in DMF (20 mL). The mixture was heated to 110 ° C and stirred for 1 h, then acetic acid (2.8 g, 47.0 mmol) was added and stirring was continued for 16 h. Cooled to room temperature and concentrated in vacuo. The residue was dissolved in EtOAc (200 mL) and brine (100 mL×3), separated, and the organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (PE:EA=3:1) to give ethyl 2-amino-5-methylpyrazolo[1,5-a]pyrimidine-3-carboxylate (2.2 g, 85% yield) as a yellow solid. LCMS:MS(ESI)m / z 221.2[M+H] + .

[0370] Step 2) 5-methyl-2-(2,2,2-trifluoroacetamido)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid ethyl ester

[0371] Trifluoroacetic anhydride (3.8 g, 18 mmol) was added to a solution of 2-amino-5-methylpyrazolo[1,5-a]pyrimidine-3-carboxylic acid ethyl ester (2 g, 9.0 mmol) and DIEA (2.3 g, 18 mmol) in DCM (30 mL) at 0°C. The mixture was transferred to room temperature and stirred for 16 hours. The mixture was diluted with DCM (100 mL) and washed with brine (50 mL x 2). The liquids were separated, and the organic phase was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=10 / 1) to give 5-methyl-2-(2,2,2-trifluoroacetamide)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid ethyl ester (1.5 g, 47% yield) as a yellow solid. LCMS:MS(ESI)m / z 316.9[M+H] + .

[0372] Step 3) Ethyl 5-formyl-2-(2,2,2-trifluoroacetamido)pyrazolo[1,5-a]pyrimidine-3-carboxylate

[0373] Selenium dioxide (421 mg, 3.8 mmol) was added to a solution of 5-methyl-2-(2,2,2-trifluoroacetamide)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid ethyl ester (600 mg, 1.9 mmol) in 1,4-dioxane (30 mL). The mixture was heated to 100°C and stirred for 36 hours. After cooling to room temperature, it was filtered and the filtrate was concentrated in vacuo. The residue was dissolved in EtOAc (200 mL) and washed with brine (100 mL x 2). The liquids were separated, the organic phase was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography (PE / EA=2 / 1) to give 5-formyl-2-(2,2,2-trifluoroacetamide)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid ethyl ester (270 mg, 43% yield) as a yellow solid. LCMS:MS(ESI)m / z 349.0[M+H] + .

[0374] Step 4) Ethyl 5-(hydroxymethyl-d)-2-(2,2,2-trifluoroacetamido)pyrazolo[1,5-a]pyrimidine-3-carboxylate

[0375] To a solution of 5-formyl-2-(2,2,2-trifluoroacetamide)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid ethyl ester (200 mg, 0.6 mmol) in THF (3 mL) at 0°C was added sodium deuterated borohydride (11 mg, 0.3 mmol). The mixture was transferred to room temperature and stirred for 16 hours. The mixture was quenched with 10% citric acid (30 mL) and extracted with EtOAc (50 mL×2). The combined organic layers were washed with brine (30 mL x 2), dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography (PE / EA=5 / 1) to give 5-(hydroxymethyl-d)-2-(2,2,2-trifluoroacetamide)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid ethyl ester (60 mg, 29% yield) as a yellow solid. LCMS:MS(ESI)m / z 334.0[M+H] + .

[0376] Step 5) Ethyl 5-(fluoromethyl-d)-2-(2,2,2-trifluoroacetamido)pyrazolo[1,5-a]pyrimidine-3-carboxylate

[0377] To a solution of ethyl 5-(hydroxymethyl)-2-(2,2,2-trifluoroacetamido)pyrazolo[1,5-a]pyrimidine-3-carboxylate (50 mg, 0.15 mmol) in DCM (10 mL) at 0°C was added DAST (48 mg, 0.3 mmol). The mixture was transferred to room temperature and stirred for 1 hour. The mixture was diluted with DCM (50 mL) and washed with brine (30 mL x 2). The liquids were separated, the organic phase was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to give ethyl 5-(fluoromethyl-d)-2-(2,2,2-trifluoroacetamido)pyrazolo[1,5-a]pyrimidine-3-carboxylate (40 mg, 80% yield) as a yellow oil. LCMS:MS (ESI) m / z 336.1 [M+H] + .

[0378] Step 6) 2-amino-5-(fluoromethyl-d)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid

[0379] To a solution of ethyl 5-(fluoromethyl)-2-(2,2,2-trifluoroacetamide)pyrazolo[1,5-a]pyrimidine-3-carboxylate (40 mg, 0.12 mmol) in EtOH (4 mL) and H2O (1 mL) was added sodium hydroxide (10 mg, 0.24 mmol). The mixture was transferred to room temperature and stirred for 16 h. The mixture was adjusted to pH = 6-7 with 1M HCl and extracted with EtOAc (30 mL x 3). The combined organic phases were dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH = 10 / 1) to give 2-amino-5-(fluoromethyl-d)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid (20 mg, yield 80%) as a yellow solid. LCMS: MS (ESI) m / z 212.1 [M+H] + .

[0380] Step 7) 2-amino-5-(fluoromethyl-d)-N-((S)-1-(8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0381] To a solution of (S)-3-(1-aminoethyl)-8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (20 mg, 0.05 mmol) in DCM (15 mL) were added 2-amino-5-(fluoromethyl-d)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid (14 mg, 0.06 mmol), HOAt (11 mg, 0.08 mmol), EDCI (13 mg, 0.08 mmol) and DIEA (21 mg, 0.16 mmol). The mixture was warmed to 40°C and stirred for 16 h. After cooling to room temperature, the mixture was diluted with DCM (50 mL) and washed with brine (30 mL x 2). The separated organic layer was dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=15 / 1) to give 2-amino-5-(fluoromethyl-d)-N-((S)-1-(8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (0.8 mg, 2.6% yield) as a white solid. LCMS: MS (ESI) m / z 562.0 [M+H] + . 1 H NMR (400MHz, CD3OD) δ8.67(d,J=7.0Hz,1H),7.77(s,1H),7.59(s,1H),7.58-7.48(m,4H),7.47-7.39(m,3H),7.35(d,J=8.8H z, 1H), 7.01 (d, J = 7.0Hz, 1H), 6.80 (s, 1H), 5.39 (d, J = 2.3Hz, 1H), 4.67 (d, J = 6.9Hz, 1H), 3.79 (s, 3H), 1.38 (d, J = 6.8Hz, 3H).

[0382] Example 11 (S)-2-amino-N-(1-(2-cyclopropyl-8-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-2-yl)ethynyl)-1-oxo-1,2-dihydroisoquinolin-3-yl)ethyl)-5-methylpyrazolo[1,5-a]pyrimidine-3-carboxamide

[0383]

[0384] Step 1) 2-Chloro-N-cyclopropyl-6-methylbenzamide

[0385] Oxalyl chloride (970 mg, 7.67 mmol) was added dropwise to a solution of 2-chloro-6-methylbenzoic acid (1.0 g, 5.9 mmol) in DCM (50 mL) and DMF (40 mg) at 0°C. The mixture was stirred for 2 h at 0°C and concentrated under reduced pressure. A solution of the above acid chloride in DCM (10 mL) was added to a solution of cyclopropylamine (340 mg, 5.9 mmol) and TEA (1.79 g, 17.7 mmol) in DCM (50 mL), and the mixture was stirred at room temperature for 5 h. The mixture was diluted with H2O (30 mL) and extracted with DCM (50 mL x 3). The combined organic phases were washed with brine (40 mL x 2), dried over Na2SO4, and concentrated under reduced pressure. The residue was purified by column chromatography (PE / EA=3 / 1) to give 2-chloro-N-cyclopropyl-6-methylbenzamide (1.2 g, 88% yield) as a yellow solid. MS(ESI):210.01[M+H] + .

[0386] Step 2) Tert-butyl (S)-(4-(3-chloro-2-(cyclopropylcarbamoyl)phenyl)-3-oxobutan-2-yl)carbamate

[0387] In a nitrogen atmosphere, at -30°C, to a solution of 2-chloro-N-cyclopropyl-6-methylbenzamide (300 mg, 1.43 mmol) in THF (20 mL) was added n-BuLi (1.6 mL, 3.72 mmol). The mixture was stirred for 1 hour. In a nitrogen atmosphere, at -30°C, to a solution of tert-butyl (S)-(1-(methoxy(methyl)amino)-1-oxopropane-2-yl)carbamate (498 mg, 2.15 mmol) in THF (20 mL) was slowly added dropwise i-PrMgCl (3.5 mL, 4.58 mmol), and the mixture was stirred for 1 hour. The solution was then slowly added to the above mixture, transferred to room temperature, and stirred for 3 h. The mixture was quenched with an aqueous solution of NH4Cl (30 mL) and extracted with EtOAc (50 mL x 3). The combined organic phase was washed with brine (10 mL x 2), dried over Na2SO4, and concentrated under reduced pressure. The residue was purified by column chromatography (PE / EA=3 / 1) to give (S)-tert-butyl (4-(3-chloro-2-(cyclopropylcarbamoyl)phenyl)-3-oxobutan-2-yl)carbamate (463 mg, yield 81%) as a yellow solid. MS (ESI): 402.9 [M+Na] + .

[0388] Step 3) (S)-3-(1-aminoethyl)-8-chloro-2-cyclopropylisoquinolin-1(2H)-one

[0389] To a solution of (S)-tert-butyl (4-(3-chloro-2-(cyclopropylcarbamoyl)phenyl)-3-oxobutan-2-yl)carbamate (200 mg, 0.53 mmol) in MeOH (10 mL) was added HCl (2.0 mL), and the mixture was heated to 80°C and stirred for 2 h. After cooling to room temperature, the mixture was concentrated under reduced pressure. The residue was diluted with H2O (10 mL), the pH value was adjusted to 7-8 with aqueous NaHCO3 solution, and then extracted with DCM (30 mL x 3), and the combined organic phases were washed with brine (30 mL x 2), dried over Na2SO4, and concentrated under reduced pressure. The residue was purified by column chromatography (DCM / MeOH=95 / 5) to give (S)-3-(1-aminoethyl)-8-chloro-2-cyclopropylisoquinolin-1(2H)-one (130 mg, 85% yield) as a yellow solid. MS(ESI):263.0[M+H] + .

[0390] Step 4) (S)-3-(1-aminoethyl)-2-cyclopropyl-8-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-2-yl)ethynyl)isoquinolin-1(2H)-one To a solution of (S)-3-(1-aminoethyl)-8-chloro-2-cyclopropylisoquinolin-1(2H)-one (50 mg, 0.19 mmol) and 2-ethynyl-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole (33 mg, 0.25 mmol) in MeCN (20 mL) under nitrogen atmosphere were added X-Phos (45 mg, 0.09 mmol), Pd2(dba)3 (43 mg, 0.05 mmol) and K3PO4 (121 mg, 0.57 mmol), and the mixture was warmed to 80°C and stirred for 5 h. After cooling to room temperature, the mixture was concentrated in vacuo. The residue was diluted with EtOAc (100 mL) and washed with brine (50 mL×2). The liquids were separated, and the organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=91 / 9) to give (S)-3-(1-aminoethyl)-2-cyclopropyl-8-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-2-yl)ethynyl)isoquinolin-1(2H)-one (33 mg, 45.3% yield) as a yellow oil. MS (ESI): 359.0 [M+H] + .

[0391] Step 5) (S)-2-amino-N-(1-(2-cyclopropyl-8-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-2-yl)ethynyl)-1-oxo-1,2-dihydroisoquinolin-3-yl)ethyl)-5-methylpyrazolo[1,5-a]pyrimidine-3-carboxamide

[0392] To a solution of (S)-3-(1-aminoethyl)-2-cyclopropyl-8-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-2-yl)ethynyl)isoquinolin-1(2H)-one (33 mg, 0.09 mmol) and 2-amino-5-methylpyrazolo[1,5-a]pyrimidine-3-carboxylic acid (18 mg, 0.09 mmol) in DCM (20 mL) were added DIEA (36 mg, 0.28 mmol), EDCI (26 mg, 0.14 mmol) and HOAT (19 mg, 0.14 mmol), and the mixture was warmed to 40°C and stirred for 16 hours. After cooling to room temperature, the mixture was diluted with DCM (100 mL) and washed with brine (40 mL x 2). The layers were separated, and the organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by preparative HPLC (-Gemini-C18 150x 21.2mm, 5um: ACN--H2O (0.1% FA), 40%-60%) to give (S)-2-amino-N-(1-(2-cyclopropyl-8-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-2-yl)ethynyl)-1-oxo-1,2-dihydroisoquinolin-3-yl)ethyl)-5-methylpyrazolo[1,5-a]pyrimidine-3-carboxamide (5.5 mg, 11% yield) as a white solid. MS (ESI): 532.9 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ8.79(d,J=6.9Hz,1H),8.22(d,J=7.2Hz,1H),7.60-7.53(m,3H), 6.91(d,J=6.9Hz,1H),6.62(s,1H),6.38(s,2H),6.26(s,1H),5.84-5.78(m,1H),4.22(t ,J=6.6Hz,1H),4.14-4.08(m,2H),3.01(s,1H),2.90-2.84(m,2H),2.56(s,3H),2.54(s, 1H), 1.58 (d, J = 6.7Hz, 3H), 1.11-1.03 (m, 2H), 0.91 (t, J = 7.4Hz, 1H), 0.86-0.76 (m, 2H).

[0393] Example 12 (S)-2-amino-N-(1-(8-((6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazol-2-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0394]

[0395] Step 1) 4-Chloro-N'-(diphenylmethylene)butyric acid hydrazide

[0396] 4-Chlorobutyryl chloride (2.8 g, 20 mmol) was added dropwise to a solution of (diphenylmethylene)hydrazine (3.9 g, 20 mmol) and pyridine (1.58 g, 20 mmol) in DCM (60 mL) at 0°C, and the mixture was stirred at room temperature for 16 hours. The mixture was diluted with DCM (200 mL) and washed with brine (100 mL x 2), separated, and the organic phase was dried over anhydrous Na2SO4, filtered, and concentrated to give 4-chloro-N'-(diphenylmethylene)butyric acid hydrazide (6.8 g, crude) as a white solid. MS (ESI): 301.0 [M+H] + .

[0397] Step 2) 1-((Diphenylmethylene)amino)pyrrolidin-2-one

[0398] At 0°C, NaH (73 mg, 3.0 mmol) was slowly added to a solution of 4-chloro-N'-(diphenylmethylene)butyric acid hydrazide (905 mg, 3.0 mmol) in THF (20 mL), and the mixture was stirred at room temperature for 16 hours. The mixture was diluted with EtOAc (60 mL), washed with brine (30 mL×2), separated, and the organic layer was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (PE / EA=2 / 1) to give 1-((diphenylmethylene)amino)pyrrolidin-2-one (768 mg, 92% yield) as a white solid. MS (ESI): 265.0 [M+H] + .

[0399] Step 3) 1-aminopyrrolidin-2-one hydrochloride

[0400] 2N HCl (5 mL) was added to a solution of 1-((diphenylmethylene)amino)pyrrolidin-2-one (503 mg, 1.9 mmol) in H2O (10 mL), transferred to room temperature, and stirred for 16 h. The mixture was extracted with DCM (30 mL), separated, and the aqueous phase was concentrated in vacuo to give 1-aminopyrrolidin-2-one hydrochloride (330 mg, crude) as a white solid. MS (ESI): 101.1 [M+H] + .

[0401] Step 4) Ethyl 2-imino-2-((2-oxopyrrolidin-1-yl)amino)acetate

[0402] To a solution of 1-aminopyrrolidin-2-one hydrochloride (330 mg, 2.4 mmol) in EtOH (20 mL) was added ethyl 2-ethoxy-2-iminoacetate (351 mg, 2.4 mmol), and the mixture was stirred at 40° C. for 16 hours. The mixture was concentrated in vacuo to give ethyl 2-imino-2-((2-oxopyrrolidin-1-yl)amino)acetate (450 mg, crude) as a white solid. MS (ESI): 200.0 [M+H] + .

[0403] Step 5) Ethyl 6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole-2-carboxylate

[0404] A mixture of ethyl 2-imino-2-((2-oxopyrrolidin-1-yl)amino)acetate (450 mg, 2.3 mmol) and POCl3 (10 mL) was stirred at 110 ° C for 16 h. After cooling to room temperature, it was concentrated in vacuo. The residue was diluted with EtOAc (50 mL) and aqueous sodium bicarbonate (30 mL). The liquids were separated, and the organic phase was washed with brine (30 mL x 2), dried over Na2SO4, and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=20 / 1) to give ethyl 6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole-2-carboxylate (100 mg, 25% yield) as a white solid. MS (ESI): 182.0 [M+H] + .

[0405] Step 6) (6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazol-2-yl)methanol

[0406] At 0°C, LiAlH4 (21 mg, 0.55 mmol) was slowly added to a solution of ethyl 6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole-2-carboxylate (100 mg, 0.55 mmol) in THF (10 mL). The mixture was stirred at room temperature for 16 h, quenched with 10% NaOH (0.1 mL) and water (5 mL), and then extracted with EtOAc (30 mL x 2). The combined organic phases were washed with brine (50 mL), dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to give (6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazol-2-yl)methanol (80 mg, crude) as a white solid. MS (ESI): 140.1 [M+H] + .

[0407] Step 7) 6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole-2-carbaldehyde

[0408] MnO2 (157 mg, 1.8 mmol) was added to a solution of (6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole-2-yl)methanol (50 mg, 0.36 mmol) in CHCl3 (20 mL), and the mixture was heated to 70°C and stirred for 16 h. After cooling to room temperature, the mixture was filtered and the filtrate was concentrated in vacuo to give 6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole-2-carboxaldehyde (40 mg, crude) as a white solid. MS (ESI): 138.0 [M+H] + .

[0409] Step 8) 2-ethynyl-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole

[0410] To a solution of 6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole-2-carboxaldehyde (40 mg, 0.30 mmol) in MeOH (10 mL) were added dimethyl (1-diazo-2-oxopropyl)phosphonate (85 mg, 0.44 mmol) and K2CO3 (81 mg, 0.58 mmol), and the mixture was stirred at room temperature for 16 hours and concentrated in vacuo. The residue was diluted with EtOAc (50 mL) and washed with brine (30 mL×2). The organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=20 / 1) to give 2-ethynyl-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole (18 mg, 45% yield) as a white solid. MS (ESI): 156.1 [M+Na] + .

[0411] Step 9) (S)-3-(1-aminoethyl)-8-((6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazol-2-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one

[0412] Under nitrogen atmosphere, 2-ethynyl-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole (27 mg, 0.20 mmol), K3PO4 (57 mg, 0.27 mmol), Pd2(dba)3 (31 mg, 0.03 mmol) and X-Phos (32 mg, 0.07 mmol) were added to a solution of (S)-3-(1-aminoethyl)-8-chloro-2-phenylisoquinolin-1(2H)-one (40 mg, 0.13 mmol) in CH3CN (20 mL), and the mixture was heated to 80°C and stirred for 16 h. After cooling to room temperature, the mixture was concentrated in vacuo. The residue was diluted with EtOAc (50 mL) and washed with brine (30 mL x 3). The liquids were separated, and the organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=20 / 1) to give (S)-3-(1-aminoethyl)-8-((6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazol-2-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (25 mg, 45% yield) as a white solid. MS (ESI): 395.9 [M+H] + .

[0413] Step 10) (S)-2-amino-N-(1-(8-((6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazol-2-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0414] To a solution of (S)-3-(1-aminoethyl)-8-((6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazol-2-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (25 mg, 0.06 mmol) in DCM (20 mL) were added 2-aminopyrazolo[1,5-a]pyrimidine-3-carboxylic acid (12 mg, 0.06 mmol), EDCI (19 mg, 0.09 mmol), HOAT (13 mg, 0.09 mmol) and DIEA (25 mg, 0.19 mmol), and the mixture was warmed to 40°C and stirred for 16 h. After cooling to room temperature, the mixture was diluted with DCM (40 mL) and washed with brine (30 mL x 2). The layers were separated, and the organic phase was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by preparative HPLC (Gemini-C18 150 x 21.2 mm, 5 um, ACN--H2O (0.1% FA) 20%-30%) to give (S)-2-amino-N-(1-(8-((6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazol-2-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (16.1 mg, yield 45%) as a white solid. MS (ESI): 555.9 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.93 (dd, J=6.7, 1.6Hz, 1H), 8.56 (dd, J=4.5, 1.6Hz, 1H ),8.02(d,J=6.7Hz,1H),7.75-7.66(m,3H),7.58-7.46(m,4H),7.41-7.37(m,1 H),7.02(dd,J=6.7,4.5Hz,1H),6.78(s,1H),6.44(s,2H),4.60-4.55(m,1H),4 .11-4.06(m,2H),2.85-2.80(m,2H),2.65-2.58(m,2H),1.36(d,J=6.8Hz,3H).

[0415] Example 13 (S)-2-amino-N-(1-(8-((2,3-dihydropyrazolo[5,1-b]oxazol-6-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0416]

[0417] Step 1) Ethyl 2,3-dihydropyrazolo[5,1-b]oxazole-6-carboxylate

[0418] 1,2-Dibromoethane (6g, 32mmol) and potassium carbonate (2.5g, 17.68mmol) were added to a solution of ethyl 5-hydroxy-1H-pyrazole-3-carboxylate (2g, 12.8mmol) in CH3CN (10mL), and the mixture was stirred at 65°C for 36h. After cooling to room temperature, the mixture was concentrated in vacuo. The residue was diluted with EtOAc (200mL) and washed with water (100mL) and brine (100mL), respectively. The liquids were separated, and the organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=10 / 1) to give ethyl 2,3-dihydropyrazolo[5,1-b]oxazole-6-carboxylate (1.8g, 77% yield) as a white solid. LCMS:MS(ESI)m / z 183.1[M+H] + .

[0419] Step 2) (2,3-dihydropyrazolo[5,1-b]oxazol-6-yl)methanol

[0420] At 0°C, LiAlH4 (31 mg, 0.82 mmol) was added to a solution of ethyl 2,3-dihydropyrazolo[5,1-b]oxazole-6-carboxylate (100 mg, 0.55 mmol) in THF (10 mL), and the mixture was transferred to room temperature and stirred for 2 h. The mixture was quenched with 10% NaOH (0.3 mL) and water (1 mL), then diluted with ethyl acetate (100 mL), separated, washed with brine (50 mL), and the organic phase was dried over anhydrous Na2SO4, filtered, and concentrated to give (2,3-dihydropyrazolo[5,1-b]oxazol-6-yl)methanol (60 mg, yield 78%) as a white solid. LCMS: MS (ESI) m / z 141.2 [M+H] + .

[0421] Step 3) 2,3-dihydropyrazolo[5,1-b]oxazole-6-carbaldehyde

[0422] MnO2 (155 mg, 1.78 mmol) was added to a solution of (2,3-dihydropyrazolo[5,1-b]oxazol-6-yl)methanol (50 mg, 0.36 mmol) in chloroform (10 mL), and the mixture was heated to 62 ° C and stirred for 36 h. After cooling to room temperature, it was filtered. The filter cake was washed twice with DCM (10 mL), and the filtrate was concentrated in vacuo to give 2,3-dihydropyrazolo[5,1-b]oxazole-6-carboxaldehyde (40 mg, 81% yield) as a white solid. LCMS: MS (ESI) m / z 139.1 [M+H] + .

[0423] Step 4) 6-ethynyl-2,3-dihydropyrazolo[5,1-b]oxazole

[0424] To a solution of 2,3-dihydropyrazolo[5,1-b]oxazole-6-carboxaldehyde (40 mg, 0.29 mmol) in MeOH (10 mL) were added dimethyl (1-diazo-2-oxopropyl)phosphonate (83 mg, 0.43 mmol) and potassium carbonate (81 mg, 0.58 mmol), and the mixture was stirred at room temperature for 16 h. The mixture was concentrated under reduced pressure. The residue was dissolved in EtOAc (100 mL) and washed with brine (50 mL). The liquids were separated, the organic phase was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to give 6-ethynyl-2,3-dihydropyrazolo[5,1-b]oxazole (20 mg, yield 52%) as a yellow solid. LCMS:MS (ESI) m / z 135.2 [M+H] + .

[0425] Step 5) (S)-3-(1-aminoethyl)-8-((2,3-dihydropyrazolo[5,1-b]oxazol-6-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one

[0426] To a solution of (S)-3-(1-aminoethyl)-8-chloro-2-phenylisoquinolin-1(2H)-one (30 mg, 0.1 mmol) in CH3CN (10 mL) were added 6-ethynyl-2,3-dihydropyrazolo[5,1-b]oxazole (20 mg, 0.15 mmol), Pd2(dba)3 (9 mg, 0.01 mmol), X-phos (5 mg, 0.01 mmol) and K3PO4 (32 mg, 0.15 mmol), and the mixture was heated to 80°C and stirred for 4 h. After cooling to room temperature, the mixture was concentrated in vacuo. The residue was dissolved in DCM (50 mL) and washed with brine (30 mL). The liquids were separated, and the organic phase was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=10 / 1) to give (S)-3-(1-aminoethyl)-8-((2,3-dihydropyrazolo[5,1-b]oxazol-6-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (20 mg, 50% yield) as a yellow solid. LCMS: MS (ESI) m / z 396.8 [M+H] + .

[0427] Step 6) (S)-2-amino-N-(1-(8-((2,3-dihydropyrazolo[5,1-b]oxazol-6-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0428] To a solution of (S)-3-(1-aminoethyl)-8-((2,3-dihydropyrazolo[5,1-b]oxazol-6-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (20 mg, 0.05 mmol) and 2-aminopyrazolo[1,5-a]pyrimidine-3-carboxylic acid (11 mg, 0.06 mmol) in DCM (20 mL) were added HOAt (10 mg, 0.07 mmol), EDCI (15 mg, 0.07 mmol) and DIEA (20 mg, 0.15 mmol), and the mixture was warmed to 40°C and stirred for 16 h. After cooling to room temperature, the mixture was diluted with DCM (50 mL) and washed with brine (30 mL). The layers were separated, and the organic phase was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography (DCM:MeOH=10:1) to give (S)-2-amino-N-(1-(8-((2,3-dihydropyrazolo[5,1-b]oxazol-6-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (9.5 mg, 33% yield) as a white solid. LCMS: MS (ESI) m / z 556.9 [M+H] + . 1 H NMR (400MHz, CD3OD) δ8.69(dd,J=6.8,1.6Hz,1H),8.50(dd,J=4.5,1.6Hz,1H),8.21(d,J=7.0Hz,1H),7.63(dd,J=14.4,8.2,6.9,2.0Hz,4H),7.53-7. 38(m,4H),6.96(dd,J=6.8,4.5Hz,1H),6.88(s,1H),5.68(s,1H),5.10-5. 02(m,2H),4.76(q,J=6.8Hz,1H),4.32-4.24(m,2H),1.46(d,J=6.8Hz,3H).

[0429] Example 14 (S)-2-amino-N-(1-(8-((2,3-dihydropyrazolo[5,1-b]oxazol-7-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0430]

[0431] Step 1) 2,3-Dihydropyrazolo[5,1-b]oxazole

[0432] 1,2-Dibromoethane (19 g, 0.09 mol) and K2CO3 (6.58 g, 0.047 mol) were added to a solution of 1,2-dihydropyrazol-3-one (2.0 g, 0.02 mol) in CH3CN (20 mL), and the mixture was heated to 80 ° C and stirred for 16 h. After cooling to room temperature, it was filtered and the filtrate was concentrated in vacuo. The residue was diluted with DCM (100 mL) and washed with brine (50 mL). The liquids were separated, the organic phase was dried over Na2SO4, and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=20 / 1) to give 2,3-dihydropyrazolo[5,1-b]oxazole (1.0 g, 34.45% yield) as a yellow oil. MS (ESI): 111.2 [M+H] + .

[0433] Step 2) 7-bromo-2,3-dihydropyrazolo[5,1-b]oxazole

[0434] NBS (873 mg, 4.9 mmol) was added to a solution of 2,3-dihydropyrazolo[5,1-b]oxazole (450 mg, 4.09 mmol) in CH3CN (10 mL), and the mixture was stirred for 5 hours. The mixture was diluted with water (20 mL) and then extracted with DCM (30 mL×3). The combined organic phase was washed with brine (30 mL x 2), dried over Na2SO4, and concentrated in vacuo. The residue was purified by column chromatography (PE:EtOAc=2:1) ​​to give 7-bromo-2,3-dihydropyrazolo[5,1-b]oxazole (500 mg, 51.79% yield) as a white solid. MS (ESI): 189.1 [M+H] + .

[0435] Step 3) 7-((triisopropylsilyl)ethynyl)-2,3-dihydropyrazolo[5,1-b]oxazole

[0436] Under nitrogen atmosphere, ethynyl triisopropylsilane (695 mg, 3.8 mmol), tripotassium phosphate (2 g, 9.5 mmol), tris(dibenzylideneacetone)dipalladium (581 mg, 0.64 mmol) and X-phos (302 mg, 0.64 mmol) were added to a solution of 7-bromo-2,3-dihydropyrazolo[5,1-b]oxazole (600 mg, 3.17 mmol) in CH3CN (20 mL), and the mixture was heated to 80 ° C and stirred for 16 hours. After cooling to room temperature, it was concentrated in vacuo. The residue was diluted with DCM (60 mL) and washed with brine (30 mL x 2). The liquid was separated, and the organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (PE:EtOAc=2:1) ​​to give 7-((triisopropylsilyl)ethynyl)-2,3-dihydropyrazolo[5,1-b]oxazole (300 mg, yield 26.03%) as a yellow oil. MS (ESI): 291.1 [M+H] + .

[0437] Step 4) 7-ethynyl-2,3-dihydropyrazolo[5,1-b]oxazole

[0438] TBAF (1 mL, 1M THF solution) was added to a solution of 7-((triisopropylsilyl)ethynyl)-2,3-dihydropyrazolo[5,1-b]oxazole (300 mg, 1.03 mmol) in THF (20 mL), and the mixture was stirred for 2 h. The mixture was diluted with EtOAc (30 mL) and washed with brine (20 mL×2). The liquids were separated, and the organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (DCM:MeOH=10:1) to give 7-ethynyl-2,3-dihydropyrazolo[5,1-b]oxazole (60 mg, 34.64% yield) as a colorless oil. MS (ESI): 135.1[M+H] + .

[0439] Step 5) (S)-3-(1-aminoethyl)-8-((2,3-dihydropyrazolo[5,1-b]oxazol-7-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one

[0440] Under nitrogen atmosphere, tris(dibenzylideneacetone)dipalladium (9 mg, 0.01 mmol), X-phos (5 mg, 0.01 mmol) and K3PO4 (43 mg, 0.2 mmol) were added to a solution of (S)-3-(1-aminoethyl)-8-chloro-2-phenylisoquinolin-1(2H)-one (30 mg, 0.1 mmol) and 7-ethynyl-2,3-dihydropyrazolo[5,1-b]oxazole (20 mg, 0.15 mmol) in acetonitrile (5 mL), and the mixture was heated to 80°C and stirred for 16 h. After cooling to room temperature, it was concentrated in vacuo. The residue was diluted with DCM (30 mL) and washed with brine (30 mL). The organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=10:1) to give (S)-3-(1-aminoethyl)-8-((2,3-dihydropyrazolo[5,1-b]oxazol-7-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (23 mg, 60.0% yield) as a yellow solid. MS (ESI): 398.0 [M+H] + .

[0441] Step 6) (S)-2-amino-N-(1-(8-((2,3-dihydropyrazolo[5,1-b]oxazol-7-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0442] To a solution of (S)-3-(1-aminoethyl)-8-((2,3-dihydropyrazolo[5,1-b]oxazol-7-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (20 mg, 0.05 mmol) and 2-aminopyrazolo[1,5-a]pyrimidine-3-carboxylic acid (11 mg, 0.06 mmol) in DCM (5 mL) were added HOAT (10 mg, 0.075 mmol), EDCI (14 mg, 0.075 mmol) and DIEA (13 mg, 0.1 mmol), and the mixture was warmed to 40°C and stirred for 6 h. After cooling to room temperature, the mixture was diluted with DCM (40 mL) and washed with brine (20 mL). The layers were separated, and the organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=10 / 1) to give (S)-2-amino-N-(1-(8-((2,3-dihydropyrazolo[5,1-b]oxazol-7-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (5.3 mg, 18.49% yield) as a yellow solid. MS (ESI): 557.0 [M+H] + . 1H NMR(400MHz,)δ8.89(dd,J=6.7,1.6Hz,1H),8.51(dd,J=4.5,1.6Hz,1H),7.9 6(d,J=6.7Hz,1H),7.61-7.51(m,3H),7.49-7.43(m,3H),7.43-7.39(m,2H), 7.35-7.30(m,1H),6.98(dd,J=6.7,4.5Hz,1H),6.68(s,1H),6.40(s,2H),5. 14-5.07(m,2H),4.53-4.48(m,1H),4.29-4.22(m,2H),1.30(t,J=3.4Hz,3H).

[0443] Example 15 (S)-2-amino-N-(1-(5-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)ethynyl)-4-oxo-3-phenyl-3,4-dihydroquinazolin-2-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0444]

[0445] Step 1) (S)-2-(1-aminoethyl)-5-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)ethynyl)-3-phenylquinazolin-4(3H)-one

[0446] Under nitrogen atmosphere, K3PO4 (62 mg, 0.3 mmol), Pd2(dba)3 (34 mg, 0.03 mmol) and X-Phos (35 mg, 0.08 mmol) were added to a solution of (S)-2-(1-aminoethyl)-5-bromo-3-phenylquinazolin-4(3H)-one (50 mg, 0.15 mmol) and 3-ethynyl-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole (29 mg, 0.22 mmol) in CH3CN (20 mL), and the mixture was heated to 80°C and stirred for 16 h. After cooling to room temperature, it was concentrated in vacuo. The residue was diluted with EtOAc (50 mL) and washed with brine (30 mL x 3). The liquid was separated, and the organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=20 / 1) to give (S)-2-(1-aminoethyl)-5-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)ethynyl)-3-phenylquinazolin-4(3H)-one (20 mg, 35% yield) as a white solid. MS (ESI): 395.9 [M+H] + .

[0447] Step 2) (S)-2-amino-N-(1-(5-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)ethynyl)-4-oxo-3-phenyl-3,4-dihydroquinazolin-2-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0448] To a solution of (S)-2-(1-aminoethyl)-5-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)ethynyl)-3-phenylquinazolin-4(3H)-one (20 mg, 0.05 mmol) and 2-aminopyrazolo[1,5-a]pyrimidine-3-carboxylic acid (10 mg, 0.05 mmol) in DCM (20 mL) were added EDCI (15 mg, 0.07 mmol), HOAT (11 mg, 0.07 mmol) and DIEA (20 mg, 0.15 mmol), and the mixture was warmed to 40°C and stirred for 16 h. The mixture was diluted with DCM (50 mL) and washed with brine (30 mL x 2), separated, and the organic phase was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by preparative HPLC (Gemini-C18 150 x 21.2 mm, 5 um, ACN--H2O (0.1% FA), gradient 20%-30%) to give (S)-2-amino-N-(1-(5-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)ethynyl)-4-oxo-3-phenyl-3,4-dihydroquinazolin-2-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (6.2 mg, yield 23%) as a white solid. MS (ESI): 555.8 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.92(dd,J=6.7,1.7Hz,1H),8.72(d,J=7.4Hz,1H),8.65(dd,J =4.5,1.6Hz,1H),7.79(d,J=7.6Hz,1H),7.69(dd,J=8.1,1.2Hz,1H),7.65-7.58(m,6H ),7.53(d,J=7.5Hz,1H),7.04-7.01(m,1H),6.49-6.42(m,2H),4.74(t,J=7.0Hz,1H), 4.08(t,J=7.2Hz,2H), 2.88(t,J=7.2Hz,2H), 2.57-2.54(m,2H), 1.32(d,J=6.6Hz,3H).

[0449] Example 16 (S)-2-amino-N-(1-(7-fluoro-8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl))ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0450]

[0451] Step 1) (S)-3-(1-aminoethyl)-7-fluoro-8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one

[0452] Under nitrogen atmosphere, to a solution of (S)-3-(1-aminoethyl)-8-chloro-7-fluoro-2-phenylisoquinolin-1(2H)-one (304 mg, 0.96 mmol) and 4-ethynyl-1-methyl-1H-pyrazole (153 mg, 1.44 mmol) in MeCN (50 mL) were added X-Phos (183 mg, 0.38 mmol), Pd2(dba)3 (176 mg, 0.20 mmol) and K3PO4 (611 mg, 2.88 mmol), and the mixture was heated to 80°C and stirred for 5 h. After cooling to room temperature, it was concentrated in vacuo. The residue was diluted with EtOAc (130 mL), washed with brine (60 mL x 2), separated, and the organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=91 / 9) to give (S)-3-(1-aminoethyl)-7-fluoro-8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (210 mg, 53.8% yield) as a yellow oil. MS (ESI): 386.9 [M+H] + .

[0453] Step 2) (S)-2-amino-N-(1-(7-fluoro-8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl))ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0454] To a solution of (S)-3-(1-aminoethyl)-7-fluoro-8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (120 mg, 0.31 mmol) and 2-amino-5-methylpyrazolo[1,5-a]pyrimidine-3-carboxylic acid (61 mg, 0.34 mmol) in DCM (20 mL) were added DIEA (160 mg, 1.24 mmol), EDCI (89 mg, 0.47 mmol) and HOAT (63 mg, 0.47 mmol), and the mixture was warmed to 40°C and stirred for 16 h. After cooling to room temperature, the mixture was diluted with DCM (100 mL) and washed with brine (60 mL x 2). The layers were separated, and the organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by preparative HPLC (-Xbridge-C18 150x 19mm, 5um, ACN-H2O (0.05% NH4OH), 40%-45%) to give (S)-2-amino-N-(1-(7-fluoro-8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl))ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (55 mg, 32.4% yield) as a white solid. MS (ESI): 546.8 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.93 (dd, J=6.7, 1.6Hz, 1H), 8.55 (dd, J=4.5, 1.6Hz, 1H ),8.05(s,1H),7.99(d,J=6.6Hz,1H),7.75-7.65(m,2H),7.63-7.55(m,2H), 7.53-7.46(m,3H),7.41-7.36(m,1H),7.02(dd,J=6.7,4.5Hz,1H),6.79(s, 1H), 6.43 (s, 2H), 4.54 (t, J = 6.7Hz, 1H), 3.82 (s, 3H), 1.35 (d, J = 6.8Hz, 3H).

[0455] Example 17 (S)-2-amino-6-fluoro-N-(1-(8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0456]

[0457] Step 1) (E)-2-Fluoro-3-hydroxyacrolein

[0458] PTSA (2.1 g, 0.01 mol) was added to a solution of tetramethoxypropane (2.0 g, 0.01 mol) in water (10 mL), and the mixture was stirred at room temperature for 2 h. Selective fluorine reagent (Selectflour, 5.9 g, 0.02 mol) was added to the mixture, and stirring was continued for 16 h. The resulting solution was used directly in the next step. MS (ESI): 91.1 [M+H] + .

[0459] Step 2) Ethyl 2-amino-6-fluoropyrazolo[1,5-a]pyrimidine-3-carboxylate

[0460] DMSO (10 mL) was slowly added to a solution of (E)-2-fluoro-3-hydroxyacrolein (2 mL). After the mixture was stirred for 10 minutes, ethyl 3,5-diamino-1H-pyrazole-4-carboxylate (283 mg, 1.67 mmol) was added. The mixture was warmed to 85 ° C and stirred for 4 h. After cooling to room temperature, the mixture was extracted with EtOAc (30 mL x 3). The liquid was separated, the organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=20:1) to give ethyl 2-amino-6-fluoropyrazolo[1,5-a]pyrimidine-3-carboxylate (100 mg, 40.3% yield) as a yellow solid. MS (ESI): 225.0 [M+H] +.

[0461] Step 3) 2-amino-6-fluoropyrazolo[1,5-a]pyrimidine-3-carboxylic acid

[0462] To a solution of ethyl 2-amino-6-fluoropyrazolo[1,5-a]pyrimidine-3-carboxylate (100 mg, 0.45 mmol) in MeOH (5 mL) was added a solution of NaOH (89 mg, 2.23 mmol) in H2O (2 mL), and the mixture was warmed to 40°C and stirred for 8 h. After cooling to room temperature, it was concentrated in vacuo. The mixture was acidified to pH=5-6 with 1N HCl, filtered, and the filter cake was washed with water (5 mL). The filtrate was concentrated in vacuo to give 2-amino-6-fluoropyrazolo[1,5-a]pyrimidine-3-carboxylic acid (60 mg, 54.67% yield) as a yellow solid. MS (ESI): 197.1 [M+H] + .

[0463] Step 4) (S)-2-amino-6-fluoro-N-(1-(8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0464] HOAT (5 mg, 0.32 mmol), EDCI (62 mg, 0.32 mmol) and DIEA (55 mg, 0.43 mmol) were added to a solution of 2-amino-6-fluoropyrazolo[1,5-a]pyrimidine-3-carboxylic acid (42 mg, 0.21 mmol) and (S)-3-(1-aminoethyl)-8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (95 mg, 0.26 mmol) in DCM (5 mL), and the mixture was warmed to 40°C and stirred for 16 h. The mixture was diluted with DCM (20 mL) and washed with brine (20 mL). The liquid was separated, and the organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by preparative HPLC (ACN--H2O (0.1% FA) gradient: 40%-70%) to give (S)-2-amino-6-fluoro-N-(1-(8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (16 mg, 14.11% yield) as a yellow solid. MS (ESI): 547.0 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.41(dd,J=4.8,2.6Hz,1H),8.78(d,J=2.4Hz,1H),8.00(s,1H),7.84(d,J=6.8Hz,1H),7.68-7.55(m ,5H),7.53-7.44(m,3H),7.39-7.35(m,1H),6.75(s,1H),6.50(s,2H),4.59-4.52(m,1H),3.82(s,3H),1.35(d,J=6.8Hz,3H).

[0465] Example 18 (S)-N-(1-(8-((1-acetylazetidin-3-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)-2-aminopyrazolo[1,5-a]pyrimidine-3-carboxamide

[0466]

[0467] Step 1) tert-butyl 3-ethynylazetidine-1-carboxylate

[0468] K2CO3 (370 mg, 2.69 mmol) and dimethyl (1-diazo-2-oxopropyl)phosphonate (309 mg, 1.61 mmol) were added to a solution of tert-butyl 3-formylazetidine-1-carboxylate (200 mg, 1.07 mmol) in MeOH (10 mL), and the mixture was transferred to room temperature and stirred for 16 h. The mixture was concentrated in vacuo, diluted with EtOAc (100 mL), and washed with brine (40 ml × 2). The liquid was separated, the organic phase was dried over Na2SO4, and concentrated in vacuo. The residue was separated and purified by column chromatography (DCM / MEOH=20 / 1) to give tert-butyl 3-ethynylazetidine-1-carboxylate (180 mg, 83% yield) as a brown oil. MS (ESI): 126.0 [M-55] + .

[0469] Step 2) 3-ethynylazetidine

[0470] To a solution of tert-butyl 3-ethynylazetidine-1-carboxylate (180 mg, 0.99 mmol) in DCM (10 mL) was added TFA (5 mL), and the mixture was stirred at room temperature for 2 h. The mixture was concentrated in vacuo to give 3-ethynylazetidine (100 mg, 99.8% yield) as a brown oil. MS (ESI): 82.1 [M+H] + .

[0471] Step 3) 1-(3-ethynylazetidin-1-yl)ethan-1-one

[0472] To a solution of 3-ethynylazetidine (118 mg, 1.45 mmol) and DIEA (1.8 g, 14.5 mmol) in DCM (20 mL) was added acetyl chloride (228 mg, 2.91 mmol) at 0°C, and the mixture was stirred at room temperature for 3 hours. The mixture was diluted with DCM (50 mL) and washed with brine (20 mL x 2). The liquids were separated, the organic phase was dried over Na2SO4, and concentrated in vacuo. The residue was separated and purified by column chromatography (DCM / MeOH=20 / 1) to give 1-(3-ethynylazetidine-1-yl)ethane-1-one (150 mg, 75.36% yield) as a yellow oil. MS (ESI): 124.1 [M+H] + .

[0473] Step 4) (S)-8-((1-acetylazetidin-3-yl)ethynyl)-3-(1-aminoethyl)-2-phenylisoquinolin-1(2H)-one

[0474] X-Phos (38 mg, 0.08 mmol), Pd2(dba)3 (37 mg, 0.04 mmol) and K3PO4 (128 mg, 0.60 mmol) were added to a solution of (S)-3-(1-aminoethyl)-8-chloro-2-phenylisoquinolin-1(2H)-one (60 mg, 0.20 mmol) and 1-(3-ethynylazetidin-1-yl)ethane-1-one (37 mg, 0.30 mmol) in MeCN (30 mL), and the mixture was heated to 80°C and stirred for 5 h. After cooling to room temperature, the mixture was concentrated in vacuo. The residue was diluted with EtOAc (100 mL) and washed with brine (50 mL×2). The organic phase was separated, dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=91 / 9) to give (S)-8-((1-acetylazetidin-3-yl)ethynyl)-3-(1-aminoethyl)-2-phenylisoquinolin-1(2H)-one (47 mg, 54.6% yield) as a yellow oil. MS (ESI): 386.0 [M+H] + .

[0475] Step 5) (S)-N-(1-(8-((1-acetylazetidin-3-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)-2-aminopyrazolo[1,5-a]pyrimidine-3-carboxamide

[0476] To a solution of (S)-8-((1-acetylazetidin-3-yl)ethynyl)-3-(1-aminoethyl)-2-phenylisoquinolin-1(2H)-one and 2-aminopyrazolo[1,5-a]pyrimidine-3-carboxylic acid (22 mg, 0.12 mmol) in DCM (20 mL) were added DIEA (47 mg, 0.37 mmol), EDCI (35 mg, 0.18 mmol) and HOAT (25 mg, 0.18 mmol), and the mixture was warmed to 40°C and stirred for 16 h. After cooling to room temperature, the mixture was diluted with DCM (30 mL) and washed with brine (20 mL x 2). The liquids were separated, and the organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by preparative HPLC (Gemini-C18 150x21.2mm, 5um ACN--H2O (0.1% FA) 35%-50%) to give (S)-N-(1-(8-((1-acetylazetidin-3-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)-2-aminopyrazolo[1,5-a]pyrimidine-3-carboxamide (13.1 mg, 19.7% yield) as a white solid. MS (ESI): 546.0 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ8.93(dd,J=6.7,1.6Hz,1H),8.55(dd,J=4.5,1.6Hz,1H),8.00(d,J=6.6Hz,1H),7.64 (dd,J=7.0,4.3Hz,2H),7.60-7.53(m,2H),7.53-7.48(m,2H),7.45(d,J=7.8Hz,1H),7.37(dd,J=5.7,3.5Hz, 1H),7.02(dd,J=6.7,4.5Hz,1H),6.74(s,1H),6.43(s,2H),4.54(q,J=6.6Hz,1H),4.38(t,J=8.4Hz,1H),4.1 5-4.08(m,2H),3.79(dd,J=8.8,6.3Hz,1H),3.68(dd,J=8.9,6.2Hz,1H),1.73(s,3H),1.34(d,J=6.8Hz,3H).

[0477] Example 19 2-amino-N-((1S)-1-(2-(2-methyl-5-oxooxolan-2-yl)ethynyl)-1-oxo-2-phenylisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0478]

[0479] Step 1) 4-oxo-6-(trimethylsilyl)hex-5-ynoic acid methyl ester

[0480] To a solution of methyl 4-chloro-4-oxobutanoate (100 mg, 0.66 mmol) and trimethyl(2-(trimethylsilyl)ethynyl)silane (113 mg, 0.66 mmol) in DCM (10 mL) at 0°C was added AlCl3 (133 mg, 0.99 mmol) and the mixture was stirred for 1 hour. The mixture was quenched with 1N HCl (4 mL) and diluted with DCM (50 mL). The mixture was washed with brine (30 mL x 2), dried over Na2SO4, and concentrated in vacuo. The residue was purified by column chromatography (EA / PE=1 / 3) to give methyl 4-oxo-6-(trimethylsilyl)hex-5-ynoate (96.0 mg, 68.1% yield) as a colorless oil. MS(ESI):213.1[M+H] + .

[0481] Step 2) 5-methyl-5-(2-(trimethylsilyl)ethynyl)oxolan-2-one

[0482] MeMgCl (0.18 mL, 0.56 mmol, 3 mol / L) was added to a solution of methyl 4-oxo-6-(trimethylsilyl)hex-5-ynoate (120 mg, 0.56 mmol) in THF (10 mL) at -20 °C, and the mixture was stirred at room temperature for 16 h. The mixture was quenched with NH4Cl solution (5 mL) and extracted with EtOAc (50 mL). The organic phase was separated and washed with brine (30 mL), dried over Na2SO4, and concentrated in vacuo to give crude 5-methyl-5-[2-(trimethylsilyl)ethynyl]oxacyclopentane-2-one (105 mg, 94.62% yield) as a colorless oil. MS (ESI): 143.1 [M+H] + .

[0483] Step 3) 5-ethynyl-5-methyloxolan-2-one

[0484] To a solution of 5-methyl-5-(2-(trimethylsilyl)ethynyl)oxolan-2-one (92 mg, 0.47 mmol) in THF (10 mL) was added TBAF (0.25 mL, 1 M) at -10°C, and the mixture was stirred at room temperature for 1 hour. The mixture was diluted with EtOAc (50 mL) and washed with brine (30 mL). The liquids were separated, and the organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (EA / PE=1 / 1) to give 5-ethynyl-5-methyloxolan-2-one (25 mg, 41.25% yield) as a brown oil. 1 H NMR (400MHz, CDCl3) δ2.85-2.75(m,1H),2.63-2.49(m,2H),2.59(s,1H),2.23-2.15(m,1H),1.72(s,3H).

[0485] Step 4) 3-((1S)-1-aminoethyl)-8-(2-(2-methyl-5-oxooxolan-2-yl)ethynyl)-2-phenylisoquinolin-1-one

[0486] Under nitrogen atmosphere, K3PO4 (128 mg, 0.6 mmol), X-phos (38 mg, 0.08 mmol) and Pd2(dba)3 (37 mg, 0.04 mmol) were added to a CH3CN solution of 3-((1S)-1-aminoethyl)-8-chloro-2-phenylisoquinolin-1-one (60 mg, 0.2 mmol) and 5-ethynyl-5-methyloxacyclopentane-2-one (30 mg, 0.24 mmol), and the mixture was warmed to 80°C and stirred for 16 h. After cooling to room temperature, the mixture was concentrated in vacuo. The residue was diluted with EtOAc (100 mL) and washed with brine (30 mL x 2). The liquids were separated, and the organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (MeOH / DCM=1 / 20) to give 3-((1S)-1-aminoethyl)-8-(2-(2-methyl-5-oxooxolan-2-yl)ethynyl)-2-phenylisoquinolin-1-one (37 mg, 74% yield) as a yellow solid. MS (ESI): 387.1 [M+H] + .

[0487] Step 5) 2-amino-N-((1S)-1-(2-(2-methyl-5-oxooxolan-2-yl)ethynyl)-1-oxo-2-phenylisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0488] 3-((1S)-1-aminoethyl)-8-(2-(1-methyl-5-oxopyrrolidin-3-yl)ethynyl)-2-phenylisoquinolin-1-one (50 mg, 0.13 mmol) was added to a solution of 2-aminopyrazolo[1,5-a]pyrimidine-3-carboxylic acid (25 mg, 0.14 mmol), HOAt (26 mg, 0.19 mmol), EDCI (37 mg, 0.19 mmol) and DIEA (50 mg, 0.39 mmol) in DCM (10 mL), and the mixture was warmed to 40°C and stirred for 16 h. After cooling to room temperature, the mixture was diluted with DCM (50 mL) and washed with brine (30 mL x 2). The layers were separated, and the organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by preparative HPLC (ACN-H2O 0.1% FA, gradient 30% to 60%) to give 2-amino-N-((1S)-1-(2-(2-methyl-5-oxooxolan-2-yl)ethynyl)-1-oxo-2-phenylisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (3.1 mg, 4.25% yield) as a white solid. MS (ESI): 547.2 [M+H] + . 1H NMR (400MHz, CD3OD) δ8.72(d,J=6.8Hz,1H),8.53(d,J=4.1Hz,1H),8.25(s,1H),7.66(d,J=4.4 Hz,2H),7.64(d,J=1.7Hz,1H),7.61(d,J=8.6Hz,1H),7.51(d,J=7.1Hz,1H),7.48(s,1H),7.43( d,J=8.0Hz,1H),6.99(dd,J=6.7,4.9Hz,1H),6.90(s,1H),4.79-4.74(m,1H),2.86(d,J=5.1Hz ,1H),2.66-2.56(m,2H),2.28(dd,J=8.8,2.8Hz,1H),1.77(s,3H),1.48(dd,J=6.8,1.5Hz,3H).

[0489] Example 20 (S)-2-amino-N-(1-(8-((6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-3-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0490]

[0491] Step 1) 6,7-dihydro-5H-pyrrolo[1,2-a]imidazole-3-carbaldehyde

[0492] At -78 ° C, n-BuLi (0.88 mL, 2.22 mmol) was added to a solution of 6,7-dihydro-5H-pyrrolo[1,2-a]imidazole (200 mg, 1.85 mmol) in THF (10 mL), and the mixture was warmed to -5 ° C and stirred for 1 hour. The mixture was cooled to -75 ° C, N,N-dimethylformamide (202 mg, 2.77 mmol) was added, and the temperature was raised to 20 ° C and stirred for 2 hours. After cooling to 0-5 ° C, the reaction was quenched with saturated NH4Cl aqueous solution (40 mL) and extracted with EtOAc (60 mL×2). The combined organic layer was washed with brine (50 mL x2), dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=20 / 1) to give 6,7-dihydro-5H-pyrrolo[1,2-a]imidazole-3-carbaldehyde (200 mg, yield 79%) as a yellow oil. LCMS: MS (ESI) m / z 137.0 [M+H] + .

[0493] Step 2) 3-ethynyl-6,7-dihydro-5H-pyrrolo[1,2-a]imidazole

[0494] To a solution of 6,7-dihydro-5H-pyrrolo[1,2-a]imidazole-3-carboxaldehyde (200 mg, 1.47 mmol) in MeOH (10 mL) were added dimethyl (1-diazo-2-oxopropyl)phosphonate (423 mg, 2.2 mmol) and potassium carbonate (412 mg, 2.94 mmol), and the mixture was stirred at room temperature for 16 h. The mixture was concentrated in vacuo, the residue was diluted with EtOAc (100 mL), and washed with brine (50 mL). The liquids were separated, the organic phase was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to give 3-ethynyl-6,7-dihydro-5H-pyrrolo[1,2-a]imidazole (130 mg, 67% yield) as a yellow solid. LCMS:MS (ESI) m / z 133.1 [M+H] + .

[0495] Step 3) (S)-3-(1-aminoethyl)-8-((6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-3-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one To a solution of (S)-3-(1-aminoethyl)-8-chloro-2-phenylisoquinolin-1(2H)-one (30 mg, 0.1 mmol) and 3-ethynyl-6,7-dihydro-5H-pyrrolo[1,2-a]imidazole (20 mg, 0.15 mmol) in CH3CN (10 mL) were added Pd2(dba)3 (9 mg, 0.01 mmol), X-phos (5 mg, 0.01 mmol) and K3PO4 (32 mg, 0.15 mmol), and the mixture was heated to 80°C and stirred for 4 h. After cooling to room temperature, it was concentrated in vacuo. The residue was dissolved in DCM (50 mL) and washed with brine (30 mL). The organic phase was separated and dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=10 / 1) to give (S)-3-(1-aminoethyl)-8-((6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-3-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (40 mg, 99% yield) as a yellow solid. LCMS:MS(ESI)m / z 395.2[M+H] + .

[0496] Step 4) (S)-2-amino-N-(1-(8-((6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-3-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0497] To a solution of (S)-3-(1-aminoethyl)-8-((6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-3-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (40 mg, 0.1 mmol) and 2-aminopyrazolo[1,5-a]pyrimidine-3-carboxylic acid (18 mg, 0.1 mmol) in DCM (20 mL) were added HOAt (21 mg, 0.15 mmol), EDCI (24 mg, 0.15 mmol) and DIEA (39 mg, 0.3 mmol), and the mixture was warmed to 40°C and stirred for 16 h. After cooling to room temperature, the mixture was diluted with DCM (50 mL) and washed with brine (30 mL x 2). The layers were separated, and the organic phase was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=10 / 1) to give (S)-2-amino-N-(1-(8-((6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-3-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (10.1 mg, 18% yield) as a white solid. LCMS: MS (ESI) m / z 555.0 [M+H] + . 1 H NMR(400MHz, DMSO-d6)δ8.93(dd,J=6.7,1.6Hz,1H),8.55(dd,J=4.5,1.6Hz,1H),8.01(d, J=6.7Hz,1H),7.70-7.64(m,2H),7.64-7.55(m,2H),7.54-7.46(m,3H),7.43-7.37(m,1H), 7.21(s,1H),7.02(dd,J=6.7,4.5Hz,1H),6.76(s,1H),6.43(s,2H),4.56(t,J=6.7Hz,1H), 3.97(t,J=7.0Hz,2H), 2.79(t,J=7.4Hz,2H), 2.55(d,J=7.5Hz,2H), 1.35(d,J=6.8Hz,3H).

[0498] Example 21 (S)-2-amino-5-(methoxy-d3)-N-(1-(8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0499]

[0500] To a solution of (S)-3-(1-aminoethyl)-8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (30 mg, 0.08 mmol) and 2-amino-5-(methoxy-d3)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid (17 mg, 0.08 mmol) in DCM (20 mL) were added DIEA (31 mg, 0.24 mmol), EDCI (23 mg, 0.12 mmol) and HOAT (17 mg, 0.12 mmol), and the mixture was stirred at 40°C for 24 hours. After cooling to room temperature, the mixture was diluted with DCM (100 mL) and washed with brine (40 mL x 2). The layers were separated, and the organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by preparative HPLC (-Xbridge-C18 150x 19mm, 5um, ACN-H2O (0.1% TFA) 40%-60%) to give (S)-2-amino-5-(methoxy-d3)-N-(1-(8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (19.1 mg, 41.3% yield) as a white solid. MS (ESI): 561.8 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.74-8.64(m,1H),8.01(s,1H),7.88(d,J=6.4Hz,1H),7.68-7.44(m,8H),7.39(d,J=6.9H z,1H),6.79(s,1H),6.47(d,J=7.3Hz,1H),6.14(s,2H),4.52-4.41(m,1H),3.82(s,3H),1.36(d,J=6.6Hz,3H).

[0501] Example 22 (S)-2-amino-N-(1-(8-((6,7-dihydro-5H-pyrazolo[5,1-b][1,3]oxazin-3-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0502]

[0503] Step 1) 6,7-dihydro-5H-pyrazolo[5,1-b][1,3]oxazine

[0504] 1,3-Dibromopropane (9.1 g, 45 mmol) and K2CO3 (6.2 g, 45 mmol) were added to a solution of 2,4-dihydro-3H-pyrazol-3-one (2.5 g, 30 mmol) in DMF (50 mL), and the mixture was heated to 130 °C and stirred for 16 h. After cooling to room temperature, the mixture was diluted with H2O (30 mL) and extracted with EtOAc (50 mL x 3). The organic phases were combined, washed with brine (30 mL x 2), dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to give 6,7-dihydro-5H-pyrazolo[5,1-b][1,3]oxazine (1.9 g, crude) as a white solid. MS (ESI): 125.1 [M+H] + .

[0505] Step 2) 3-Bromo-6,7-dihydro-5H-pyrazolo[5,1-b][1,3]oxazine

[0506] To a solution of 6,7-dihydro-5H-pyrazolo[5,1-b][1,3]oxazine (1.9 g, 15.3 mmol) in DCM (40 mL) was added NBS (2.7 g, 15.3 mmol) at 20 °C, and the mixture was stirred for 16 hours. The mixture was diluted with DCM (50 mL) and washed with brine (20 mL x 2). The liquids were separated, and the organic phase was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography (PE / EA=1 / 1) to give 3-bromo-6,7-dihydro-5H-pyrazolo[5,1-b][1,3]oxazine (1.7 g, 53% yield) as a white solid. MS (ESI): 202.9 [M+H] + .

[0507] Step 3) 3-((triisopropylsilyl)ethynyl)-6,7-dihydro-5H-pyrazolo[5,1-b][1,3]oxazine

[0508] Under nitrogen atmosphere, ethynyl triisopropylsilane (1.05 g, 5.76 mmol), K3PO4 (1.6 g, 7.68 mmol), Pd2(dba)3 (880 mg, 0.96 mmol) and X-Phos (916 mg, 1.92 mmol) were added to a solution of 3-bromo-6,7-dihydro-5H-pyrazolo[5,1-b][1,3]oxazine (780 mg, 3.84 mmol) in CH3CN (20 mL), and the mixture was heated to 80°C and stirred for 16 h. Cooled to room temperature and concentrated in vacuo. The residue was diluted with EtOAc (50 mL) and washed with brine (30 mL x 3). The liquid was separated, the organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=20 / 1) to give 3-((triisopropylsilyl)ethynyl)-6,7-dihydro-5H-pyrazolo[5,1-b][1,3]oxazine (450 mg, 38% yield) as a white solid. MS (ESI): 305.0 [M+H] + .

[0509] Step 4) 3-ethynyl-6,7-dihydro-5H-pyrazolo[5,1-b][1,3]oxazine

[0510] To a solution of 3-((triisopropylsilyl)ethynyl)-6,7-dihydro-5H-pyrazolo[5,1-b][1,3]oxazine (450 mg, 1.48 mmol) in THF (10 mL) was added TBAF (3.0 mL, 1 M in THF), and the mixture was stirred at room temperature for 1 hour. The mixture was extracted with EtOAc (60 mL) and washed with brine (40 mL×2). The liquids were separated, and the organic phase was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography (PE / EA=1 / 1) to give 3-ethynyl-6,7-dihydro-5H-pyrazolo[5,1-b][1,3]oxazine (200 mg, 88% yield) as a brown solid. MS(ESI):149.0[M+H] + .

[0511] Step 5) (S)-3-(1-aminoethyl)-8-((6,7-dihydro-5H-pyrazolo[5,1-b][1,3]oxazin-3-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one

[0512] Under nitrogen atmosphere, K3PO4 (72 mg, 0.34 mmol), Pd2(dba)3 (16 mg, 0.01 mmol) and X-Phos (16 mg, 0.03 mmol) were added to a solution of 3-ethynyl-6,7-dihydro-5H-pyrazolo[5,1-b][1,3]oxazine (37.2 mg, 0.25 mmol) and (S)-3-(1-aminoethyl)-8-chloro-2-phenylisoquinolin-1(2H)-one (50 mg, 0.16 mmol) in CH3CN (20 mL), and the mixture was heated to 80°C and stirred for 16 h. After cooling to room temperature, the mixture was concentrated in vacuo. The residue was diluted with EtOAc (50 mL) and washed with brine (30 mL x 3). The liquid was separated, and the organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=20 / 1) to give (S)-3-(1-aminoethyl)-8-((6,7-dihydro-5H-pyrazolo[5,1-b][1,3]oxazin-3-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (30 mg, 43% yield) as a white solid. MS (ESI): 410.9 [M+H] + .

[0513] Step 6) (S)-2-amino-N-(1-(8-((6,7-dihydro-5H-pyrazolo[5,1-b][1,3]oxazin-3-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0514] To a solution of (S)-3-(1-aminoethyl)-8-((6,7-dihydro-5H-pyrazolo[5,1-b][1,3]oxazin-3-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (30 mg, 0.07 mmol) and 2-aminopyrazolo[1,5-a]pyrimidine-3-carboxylic acid (14 mg, 0.07 mmol) in DCM (20 mL) were added EDCI (22 mg, 0.10 mmol), HOAT (15 mg, 0.10 mmol) and DIEA (29 mg, 0.21 mmol), and the mixture was warmed to 40°C and stirred for 16 h. The mixture was diluted with DCM (50 mL) and washed with brine (30 mL x 2). The layers were separated, and the organic phase was washed with brine (50 mL), dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=20 / 1) to give (S)-2-amino-N-(1-(8-((6,7-dihydro-5H-pyrazolo[5,1-b][1,3]oxazin-3-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (27.8 mg, 66% yield) as a white solid. MS (ESI): 570.8 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ8.93(dd,J=6.7,1.6Hz,1H),8.55(dd,J=4.5,1.6Hz,1H),8 .00(d,J=6.8Hz,1H),7.63-7.55(m,3H),7.52-7.45(m,4H),7.41(s,1H),7.39-7.35 (m,1H),7.02(dd,J=6.7,4.5Hz,1H),6.72(s,1H),6.44(s,2H),4.57-4.52(m,1H), 4.38-4.34(m,2H),4.07(t,J=6.1Hz,2H),2.21-2.13(m,2H),1.35(d,J=6.8Hz,3H).

[0515] Example 23 (S)-2-amino-5-(methoxy-d3)-N-(1-(5-((1-methyl-1H-pyrazol-4-yl)ethynyl)-4-oxo-3-phenyl-3,4-dihydroquinazolin-2-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0516]

[0517] Step 1) (S)-2-(1-aminoethyl)-5-((1-methyl-1H-pyrazol-4-yl)ethynyl)-3-phenylquinazolin-4(3H)-one

[0518] K3PO4 (247 mg, 1.2 mmol), Pd2(dba)3 (53 mg, 0.058 mmol) and X-phos (55 mg, 0.12 mmol) were added to a solution of 2-((1S)-1-aminoethyl)-5-bromo-3-phenylquinazolin-4-one (200 mg, 0.58 mmol) and 4-ethynyl-1-methylpyrazole (93 mg, 0.87 mmol) in CH3CN (30 mL), and the mixture was warmed to 80°C and heated for 5 h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was diluted with EtOAc (60 mL) and washed with brine (40 mL x 2). The liquid was separated, and the organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=20 / 1) to give (S)-2-(1-aminoethyl)-5-((1-methyl-1H-pyrazol-4-yl)ethynyl)-3-phenylquinazolin-4(3H)-one (130 mg, 54.5% yield) as a yellow solid. MS (ESI): 370.1 [M+H] + .

[0519] Step 2) 2-amino-5-(methoxy-d3)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid

[0520] Na (29 mg, 1.24 mmol) was added to CD3OD (3 mL), and the mixture was stirred at 20 °C for 1 hour. Then 2-amino-5-chloropyrazolo[1,5-a]pyrimidine-3-carboxylic acid ethyl ester (100 mg, 0.41 mmol) was added. The mixture was stirred for 2 h. The mixture was quenched with H2O (1 mL), warmed to 40 °C, and heated for 16 h. The mixture was concentrated in vacuo, the residue was diluted with water (5 mL), and then acidified to pH = 4-5 with 1N HCl. Filter, wash the filter cake with water (2 mL), and concentrate the filtrate under reduced pressure to give 2-amino-5-(methoxy-d3)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid (90 mg, 92.3% yield) as a light solid. MS (ESI): 237.1 [M+H] + . 1 HNMR(400MHz,DMSO-d6)δ8.66-8.56(m,1H),6.49-6.37(m,1H),6.15(s,2H).

[0521] Step 3) (S)-2-amino-5-(methoxy-d3)-N-(1-(5-((1-methyl-1H-pyrazol-4-yl)ethynyl)-4-oxo-3-phenyl-3,4-dihydroquinazolin-2-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0522] To a solution of 2-((1S)-1-aminoethyl)-5-(2-(1-methylpyrazol-4-yl)ethynyl)-3-phenylquinazolin-4-one (30 mg, 0.08 mmol) and 2-amino-5-methoxypyrazolo[1,5-a]pyrimidine-3-carboxylic acid (17 mg, 0.08 mmol) in DCM (20 mL) were added HOAt (13 mg, 0.097 mmol), DIEA (32 mg, 0.24 mmol) and EDCI (23 mg, 0.12 mmol), and the mixture was warmed to 40°C and heated for 16 h. After cooling to room temperature, the mixture was diluted with DCM (30 mL) and washed with brine (30 mL). The layers were separated, and the organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=15 / 1) to give (S)-2-amino-5-(methoxy-d3)-N-(1-(5-((1-methyl-1H-pyrazol-4-yl)ethynyl)-4-oxo-3-phenyl-3,4-dihydroquinazolin-2-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (14.1 mg, 28.7% yield) as a white solid. MS (ESI): 563.2 [M+H] + . 1 H NMR (400MHz, CD3OD) δ8.44-8.41(m,1H),7.88(s,1H),7.79(t,J=8.0Hz,1H),7.69-7.61(m,5H),7.59-7.5 6(m,1H),7.53-7.50(m,1H),6.45(d,J=7.2Hz,1H),4.93-4.91(m,1H),3.89(s,3H),1.51(d,J=6.8Hz,3H).

[0523] Example 24 (S)-2-amino-N-(1-(6-fluoro-5-((1-methyl-1H-pyrazol-4-yl)ethynyl)-4-oxo-3-phenyl-3,4-dihydroquinazoline)-2-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0524]

[0525] Step 1) 2-Bromo-3-fluoro-6-nitrobenzoic acid

[0526] To a solution of 2-bromo-3-fluorobenzoic acid (3.0 g, 13.7 mmol) in H2SO4 (15 mL) was added HNO3 (3.54 g, 20.55 mmol) dropwise at 0°C, and the mixture was stirred at room temperature for 3 hours. The mixture was diluted with ice water (100 mL) and extracted with DCM (100 mL x 3). The combined organic phases were dried over Na2SO4 and concentrated in vacuo to give 2-bromo-3-fluoro-6-nitrobenzoic acid (3.35 g, yield 83.21%) as a yellow solid. MS (ESI): 263.9 [M+H] + .

[0527] Step 2) 2-Bromo-3-fluoro-6-nitro-N-phenylbenzamide

[0528] 2-Bromo-3-fluoro-6-nitrobenzoic acid (3.35 g, 12.7 mmol) was added to SOCl2 (30 mL), and the mixture was warmed to 80 ° C and stirred for 16 hours. The mixture was then concentrated under reduced pressure to obtain the acid chloride. The above acid chloride was added to a solution of aniline (1.16 g, 12.5 mmol) and TEA (8.1 g, 62.5 mol) in DCM (50 mL) at 0 ° C. The mixture was stirred at room temperature for 5 hours, then diluted with H2O (100 mL) and extracted with DCM (150 mL x 3). The combined organic layer was washed with brine (70 mL x 2), dried over Na2SO4, and concentrated under reduced pressure. The residue was purified by column chromatography (PE / EA=3 / 1) to obtain 2-bromo-3-fluoro-6-nitro-N-phenylbenzamide (2.0 g, 43.2% yield) as a yellow solid. MS (ESI): 338.8 [M+H] + .

[0529] Step 3) 6-amino-2-bromo-3-fluoro-N-phenylbenzamide

[0530] Fe (660.0 mg, 12.0 mmol) was added to 2-bromo-3-fluoro-6-nitro-N-phenylbenzamide (400 mg, 1.2 mmol) and saturated aqueous ammonium chloride solution (10 mL), and the mixture was stirred at room temperature for 16 h. After filtration, the filtrate was diluted with H2O (100 mL) and extracted with EtOAc (50 mL×3). The combined organic layer was washed with brine (40 mL x 2), dried over Na2SO4, and concentrated under reduced pressure to give 6-amino-2-bromo-3-fluoro-N-phenylbenzamide (366 mg, 90.34% yield) as a yellow solid. MS (ESI): 310.9 [M+H] + .

[0531] Step 4) Tert-butyl (S)-(1-((3-bromo-4-fluoro-2-(phenylcarbamoyl)phenyl)amino)-1-oxopropan-2-yl)carbamate

[0532] To a solution of 6-amino-2-bromo-3-fluoro-N-phenylbenzamide (366 mg, 1.18 mmol) in DCM (50 mL) were added (tert-butyloxycarbonyl)-L-alanine (224 mg, 1.18 mmol), DIEA (611 mg, 4.74 mmol) and HATU (675 mg, 1.78 mmol), and the mixture was stirred at room temperature for 16 h. The mixture was diluted with H2O (30 mL) and extracted with DCM (50 mL x 3). The combined organic layers were washed with brine (30 mL x 2), dried over Na2SO4, and concentrated under reduced pressure. The residue was purified by column chromatography (PE / EA=3 / 1) to give tert-butyl (S)-(1-((3-bromo-4-fluoro-2-(phenylcarbamoyl)phenyl)amino)-1-oxopropan-2-yl)carbamate (448 mg, yield 70.75%) as a yellow solid. MS(ESI):501.8[M+Na] + .

[0533] Step 5) (S)-2-(1-aminoethyl)-5-bromo-6-fluoro-3-phenylquinazolin-4(3H)-one

[0534] To a solution of (S)-tert-butyl (1-((3-bromo-4-fluoro-2-(phenylcarbamoyl)phenyl)amino)-1-oxopropan-2-yl)carbamate (448 mg, 0.93 mmol) in DCM (30 mL) were added HMDS (451 mg, 2.79 mmol) and I2 (472 mg, 1.86 mmol), and the mixture was warmed to 45 ° C and stirred for 16 h. After cooling to room temperature, the mixture was diluted with aqueous Na2SO3 (20 mL) and then extracted with DCM (30 mL x 3). The combined organic layers were washed with brine (50 mL x 2), dried over Na2SO4, and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=95 / 5) to give (S)-2-(1-aminoethyl)-5-bromo-6-fluoro-3-phenylquinazolin-4(3H)-one (267 mg, 71.3% yield) as a yellow solid. MS (ESI): 361.8 [M+H] + .

[0535] Step 6) (S)-2-(1-aminoethyl)-6-fluoro-5-((1-methyl-1H-pyrazol-4-yl)ethynyl)-3-phenylquinazolin-4(3H)-one

[0536] To a solution of (S)-2-(1-aminoethyl)-5-bromo-6-fluoro-3-phenylquinazolin-4(3H)-one (60 mg, 0.17 mmol) and 4-ethynyl-1-methyl-1H-pyrazole (23 mg, 0.22 mmol) in CH3CN (20 mL) were added X-Phos (39 mg, 0.083 mmol), Pd2(dba)3 (38 mg, 0.041 mmol) and K3PO4 (105 mg, 0.50 mmol), and the mixture was warmed to 80°C and stirred for 5 h. After cooling to room temperature, the mixture was concentrated in vacuo. The residue was diluted with EtOAc (60 mL), washed with brine (30 mL x 2), dried over Na2SO4, and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=91 / 9) to give (S)-2-(1-aminoethyl)-6-fluoro-5-((1-methyl-1H-pyrazol-4-yl)ethynyl)-3-phenylquinazolin-4(3H)-one (49 mg, 69% yield) as a yellow oil. MS (ESI): 388.0 [M+H] + .

[0537] Step 7) (S)-2-amino-N-(1-(6-fluoro-5-((1-methyl-1H-pyrazol-4-yl)ethynyl)-4-oxo-3-phenyl-3,4-dihydroquinazolin)-2-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0538] To a solution of (S)-2-(1-aminoethyl)-6-fluoro-5-((1-methyl-1H-pyrazol-4-yl)ethynyl)-3-phenylquinazolin-4(3H)-one (49 mg, 0.13 mmol) and 2-amino-5-methylpyrazolo[1,5-a]pyrimidine-3-carboxylic acid (25 mg, 0.14 mmol) in DCM (20 mL) were added DIEA (65 mg, 0.51 mmol), EDCI (36 mg, 0.19 mmol) and HOAT (26 mg, 0.19 mmol), and the mixture was warmed to 40°C and stirred for 16 h. After cooling to room temperature, the mixture was diluted with DCM (70 mL) and washed with brine (40 mL x 2). The liquids were separated, and the organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by preparative HPLC (-Gemini-C18 150x 21.2mm, 5um: ACN--H2O (0.1% FA), 30%-60%) to give (S)-2-amino-N-(1-(6-fluoro-5-((1-methyl-1H-pyrazol-4-yl)ethynyl)-4-oxo-3-phenyl-3,4-dihydroquinazolin)-2-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (2.95 mg, 4.27% yield) as a white solid. MS (ESI): 547.8 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.92(dd,J=6.7,1.6Hz,1H),8.70(d,J=7.4Hz,1H),8.64(dd,J=4.5,1.6Hz,1H),8.09(s,1H),7.85-7.74(m,2H),7 .67-7.57(m,5H),7.54(d,J=7.9Hz,1H),7.03(dd,J=6.7,4.5Hz,1H),6.45(s,2H),4.79-4.69(m,1H),3.83(s,3H),1.32(d,J=6.7Hz,3H).

[0539] Example 25 (S)-2-amino-N-(1-(5-((6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-3-yl)ethynyl)-4-oxo-3-phenyl-3,4-dihydroquinazolin-2-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0540]

[0541] Step 1) 6,7-dihydro-5H-pyrrolo[1,2-a]imidazole-3-carbaldehyde

[0542] To a solution of 6,7-dihydro-5H-pyrrolo[1,2-a]imidazole (200 mg, 1.85 mmol) in THF (10 mL) was added n-BuLi (0.9 mL, 2.22 mmol, 2.4 M in THF) at -78 °C, and the mixture was warmed to -10 °C and stirred for 1 hour. The mixture was then cooled to -75 °C, N,N-dimethylformamide (203 mg, 2.8 mmol) was added, and the mixture was transferred to room temperature and stirred for 2 hours. At 0-5 °C, saturated aqueous NH4Cl solution (20 mL) was added to quench the reaction. The mixture was extracted with EtOAc (50 mL x 2), and the combined organic layers were washed with brine (50 mL x 2), dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=10 / 1) to give 6,7-dihydro-5H-pyrrolo[1,2-a]imidazole-3-carbaldehyde (200 mg, yield 79%) as a yellow oil. LCMS: MS (ESI) m / z 137.0 [M+H] + .

[0543] Step 2) 3-ethynyl-6,7-dihydro-5H-pyrrolo[1,2-a]imidazole

[0544] To a solution of 6,7-dihydro-5H-pyrrolo[1,2-a]imidazole-3-carbaldehyde (200 mg, 1.47 mmol) in MeOH (10 mL) were added dimethyl (1-diazo-2-oxopropyl)phosphonate (423 mg, 2.2 mmol) and K2CO3 (412 mg, 2.94 mmol), and the mixture was stirred at room temperature for 16 hours. The mixture was concentrated in vacuo, dissolved with EtOAc (100 mL), and washed with brine (50 mL x 2). The separated organic layer was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to give 3-ethynyl-6,7-dihydro-5H-pyrrolo[1,2-a]imidazole (130 mg, 67% yield) as a yellow solid. LCMS: MS (ESI) m / z 133.1 [M+H] + .

[0545] Step 3) (S)-2-(1-aminoethyl)-5-((6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-3-yl)ethynyl)-3-phenylquinazolin-4(3H)-one To a solution of (S)-2-(1-aminoethyl)-5-bromo-3-phenylquinazolin-4(3H)-one (50 mg, 0.15 mmol) in CH3CN (10 mL) were added 3-ethynyl-6,7-dihydro-5H-pyrrolo[1,2-a]imidazole (38 mg, 0.29 mmol), Pd2(dba)3 (13 mg, 0.015 mmol), X-phos (7 mg, 0.015 mmol) and K3PO4 (46 mg, 0.22 mmol), and the mixture was warmed to 70°C and stirred for 3 h. After cooling to room temperature, the mixture was concentrated in vacuo. The residue was dissolved in DCM (50 mL) and washed with brine (30 mL x 2). The separated organic layer was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=10 / 1) to give (S)-2-(1-aminoethyl)-5-((6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-3-yl)ethynyl)-3-phenylquinazolin-4(3H)-one (30 mg, 52% yield) as a yellow solid. LCMS:MS(ESI)m / z 396.2[M+H] + .

[0546] Step 4) (S)-2-amino-N-(1-(5-((6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-3-yl)ethynyl)-4-oxo-3-phenyl-3,4-dihydroquinazolin-2-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0547] To a solution of (S)-2-(1-aminoethyl)-5-((6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-3-yl)ethynyl)-3-phenylquinazolin-4(3H)-one (30 mg, 0.08 mmol) in DCM (15 mL) were added 2-aminopyrazolo[1,5-a]pyrimidine-3-carboxylic acid (14 mg, 0.08 mmol), [1,2,3]triazolo[4,5-b]pyridin-3-ol (16 mg, 0.11 mmol), EDCI (18 mg, 0.11 mmol) and DIEA (30 mg, 0.23 mmol), and the mixture was warmed to 40 °C and stirred for 16 h. After cooling to room temperature, the mixture was diluted with DCM (50 mL) and washed with brine (30 mL x 2). The separated organic layer was dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=10 / 1) to give (S)-2-amino-N-(1-(5-((6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-3-yl)ethynyl)-4-oxo-3-phenyl-3,4-dihydroquinazolin-2-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (6.5 mg, 15% yield) as a white solid. LCMS: MS (ESI) m / z 556.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.92(dd,J=6.7,1.6Hz,1H),8.72(d,J=7.4Hz,1H),8.65(dd,J= 4.5,1.6Hz,1H),7.82(d,J=7.7Hz,1H),7.74(d,J=1.2Hz,1H),7.67(dd,J=7.5,1.2Hz,1 H),7.63(s,2H),7.61(s,3H),7.25(s,1H),7.03(dd,J=6.7,4.5Hz,1H),6.45(s,2H),4. 78-4.71(m,1H),3.96(s,2H),2.76(d,J=7.6Hz,2H),2.54(s,2H),1.32(d,J=6.7Hz,3H).

[0548] Example 26 (S)-2-amino-N-(1-(7-fluoro-8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin)-3-yl)ethyl)-5-(methoxy-d3)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0549]

[0550] To a solution of (S)-3-(1-aminoethyl)-7-fluoro-8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (49 mg, 0.38 mmol) and 2-amino-5-(methoxy-d3)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid (20 mg, 0.10 mmol) in DCM (20 mL) were added DIEA (49 mg, 0.38 mmol), EDCI (27 mg, 0.14 mmol) and HOAT (20 mg, 0.14 mmol), and the mixture was stirred at 40° C. for 16 h. After cooling to room temperature, the mixture was diluted with DCM (50 mL) and washed with brine (30 mL×2). The separated organic layer was dried over Na2SO4 and concentrated in vacuo. The residue was purified by preparative HPLC (-Xbridge-C18 150x 19mm, 5um, ACN-H2O (0.1% TFA)) to give (S)-2-amino-N-(1-(7-fluoro-8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin)-3-yl)ethyl)-5-(methoxy-d3)pyrazolo[1,5-a]pyrimidine-3-carboxamide (27.3 mg, 50.0% yield) as a white solid. MS (ESI): 579.8 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.74-8.67(m,1H),8.05(s,1H),7.88(d,J=6.5Hz,1H),7.75-7.65(m,2H),7.61-7.46(m,5H),7. 39(d,J=7.5Hz,1H),6.83(s,1H),6.51-6.45(m,1H),6.13(s,2H),4.52-4.41(m,1H),3.83(s,3H),1.35(d,J=6.7Hz,3H).

[0551] Example 27 (S)-2-amino-N-(1-(7-fluoro-8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin)-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-5,7-d2-3-carboxamide

[0552]

[0553] Step 1) N 1 ,N 3 -Dimethoxy-N 1 ,N 3 -Dimethylmalonamide

[0554] To a solution of methoxy(methyl)amine (1.08 g, 17.7 mmol) and DIEA (4.59 g, 35.5 mmol) in DCM (20 mL) at 0°C was added malonyl chloride (1 g, 7.1 mmol), and the mixture was stirred at room temperature for 16 h. The mixture was diluted with DCM (50 mL) and washed with brine (30 mL x 3). The separated organic layer was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (EA / PE=1 / 8) to give N 1 ,N 3 -Dimethoxy-N 1 ,N 3 -Dimethylmalonamide (700 mg, yield 31%) is a colorless oil. MS (ESI): 191.1 [M+H] + .

[0555] Step 2) Malondialdehyde-1,3-d2

[0556] At 0℃, N 1 ,N 3 -Dimethoxy-N 1 ,N 3 -dimethylmalonamide (500 mg, 2.63 mmol) in THF (15 mL) was added LiAlD4 (220 mg, 5.24 mmol) in portions, and the mixture was stirred for 2 h. The mixture was then quenched with 1N deuterated hydrochloric acid (5 mmol, dissolved in D2O (5 mL)), extracted with EtOAc (10 mL), and the aqueous phase was used in the next step without further purification. MS (ESI): 75.2 [M+H] + .

[0557] Step 3) Ethyl 2-amino-5,7-d2-pyrazolo[1,5-a]pyrimidine-3-carboxylate

[0558] To a solution of 3,5-diamino-1H-pyrazole-4-carboxylic acid ethyl ester (250 mg, 0.29 mmol) in HOAc (20 mL) was added the aqueous solution of malondialdehyde-1,3-d2 from the previous step, and the mixture was heated to 90 ° C and stirred for 2 h. After cooling to room temperature, the mixture was concentrated in vacuo. The residue was diluted with EtOAc (60 mL) and washed with aqueous NaHCO3 solution (40 mL) and brine (40 mL), respectively. The separated organic layer was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (EA / PE=1 / 1) to give 2-amino-5,7-d2-pyrazolo[1,5-a]pyrimidine-3-carboxylic acid ethyl ester (110 mg, 34% yield) as a yellow solid. MS (ESI): 210.2 [M+H] + .

[0559] Step 4) 2-amino-5,7-d2-pyrazolo[1,5-a]pyrimidine-3-carboxylic acid

[0560] To ethyl 2-amino-5,7-d2-pyrazolo[1,5-a]pyrimidine-3-carboxylate (120 mg, 0.58 mmol) in MeOH (20 mL) was added NaOH (115 mg, 2.9 mmol) and H2O (5 mL), and the mixture was warmed to 40 ° C and stirred for 16 h. After cooling to room temperature, the mixture was concentrated in vacuo. The residue was diluted with water (10 mL) and adjusted to pH 5-6 with 1N FA. Filter, wash the filter cake with water, and dry in vacuo to give 2-amino-5,7-d2-pyrazolo[1,5-a]pyrimidine-3-carboxylic acid (60 mg, 54.9% yield) as a yellow solid. MS (ESI): 180.9 [M+H] + .

[0561] Step 5) (S)-2-amino-N-(1-(7-fluoro-8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin)-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-5,7-d2-3-carboxamide

[0562] To a solution of 3-((1s)-1-aminoethyl)-7-fluoro-8-(2-(1-methylpyrazol-4-yl)ethylethyl)-2-phenylmethylquinolin-1-one (43 mg, 0.11 mmol) and 2-amino-5,7-d2-pyrazolo[1,5-a]pyrimidine-3-carboxylic acid (20 mg, 0.11 mmol) in DCM (30 mL) were added HOAt (18 mg, 0.13 mmol), EDCI (32 mg, 0.17 mmol) and DIEA (43 mg, 0.33 mmol), and the mixture was warmed to 40° C. and stirred for 15 h. After cooling to room temperature, the mixture was diluted with DCM (40 mL) and washed with brine (30 mL x 2). The separated organic layer was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=15 / 1) to give (S)-2-amino-N-(1-(7-fluoro-8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin)-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-5,7-d2-3-carboxamide (26.9 mg, 44% yield) as a light yellow solid. MS (ESI): 549.1 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ8.95-8.91(m,1H),8.04(s,1H),7.99(d,J=6.4Hz,1H),7.74-7.64(m,2H),7.61(s,1H),7.59-7.55(m ,1H),7.53-7.45(m,3H),7.40-7.36(m,1H),6.79(s,1H),6.42(s,2H),4.59-4.50(m,1H),3.82(s,1H),1.35(d,J=6.8Hz,3H).

[0563] Example 28 (S)-2-amino-N-(1-(7-fluoro-8-((1-(methyl-d3)-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0564]

[0565] Step 1) 4-iodo-1-(methyl-d3)-1H-pyrazole

[0566] To a solution of 4-iodo-1H-pyrazole (2 g, 10 mmol) in DMF (10 mL) was added sodium hydride (297 mg, 12.3 mmol) at 0°C, and the mixture was stirred for 0.5 h. Then iodomethane-d3 (1.8 g, 12.3 mmol) was added, transferred to room temperature, and stirred for 16 h. The mixture was quenched with ice water (20 mL) and extracted with EtOAc (50 mL×3). The combined organic layers were washed with brine (50 mL x 3), dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography (PE / EA=2 / 1) to give 4-iodo-1-(methyl-d3)-1H-pyrazole (2 g, 91% yield) as a yellow oil. LCMS:MS (ESI) m / z 212.0[M+H] + .

[0567] Step 2) 1-(Methyl-d3)-4-((trimethylsilyl)ethynyl)-1H-pyrazole

[0568] To a solution of 4-iodo-1-(methyl-d3)-1H-pyrazole (2 g, 9.6 mmol) in TEA (30 mL) were added ethynyltrimethylsilane (1.4 g, 14.4 mmol), CuI (183 mg, 0.96 mmol), bis(triphenylphosphine)palladium(II) chloride (675 mg, 0.96 mmol) and triethylamine (9.7 g, 96 mmol), and the mixture was warmed to 90°C and stirred for 4 h. After cooling to room temperature, it was concentrated in vacuo. The residue was dissolved in DCM (50 mL), washed with water (30 mL) and brine (30 mL). The separated organic layer was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography (PE / EA=3 / 1) to give 1-(methyl-d3)-4-((trimethylsilyl)ethynyl)-1H-pyrazole (1.2 g, yield 70%) as a brown oil. LCMS:MS(ESI)m / z182.1[M+H] + .

[0569] Step 3) 4-ethynyl-1-(methyl-d3)-1H-pyrazole

[0570] To a solution of 1-(methyl-d3)-4-((trimethylsilyl)ethynyl)-1H-pyrazole (2.3 g, 12.7 mmol) in THF (20 mL) was added TBAF (13 mL, 12.7 mmol, 1 N in THF), and the mixture was stirred at room temperature for 0.5 h. The reaction mixture was diluted with EtOAc (100 mL) and washed with brine (30 mL x 2). The separated organic layer was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography (PE / EA=10 / 1) to give 4-ethynyl-1-(methyl-d3)-1H-pyrazole (800 mg, yield 87%) as a yellow solid. LCMS: MS (ESI) m / z 110.1 [M+H] + .

[0571] Step 4) (S)-3-(1-aminoethyl)-7-fluoro-8-((1-(methyl-d3)-1H-pyrazol-4-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one

[0572] To a solution of (S)-3-(1-aminoethyl)-7-fluoro-8-((1-(methyl-d3)-1H-pyrazol-4-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (150 mg, 0.47 mmol) and 4-ethynyl-1-(methyl-d3)-1H-pyrazole (77 mg, 0.71 mmol) in CH3CN (20 mL) were added Pd2(dba)3 (87 mg, 0.095 mmol), X-phos (90 mg, 0.19 mmol) and K3PO4 (150 mg, 0.71 mmol), and the mixture was warmed to 80°C and stirred for 4 h. After cooling to room temperature, it was concentrated in vacuo. The residue was dissolved in DCM (50 mL) and washed with brine (30 mL x 2). The separated organic layer was dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=10 / 1) to give (S)-3-(1-aminoethyl)-7-fluoro-8-((1-(methyl-d3)-1H-pyrazol-4-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (120 mg, 65% yield) as a brown oil. LCMS: MS (ESI) m / z 390.1 [M+H] + .

[0573] Step 5) (S)-2-amino-N-(1-(7-fluoro-8-((1-(methyl-d3)-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0574] To a solution of (S)-3-(1-aminoethyl)-7-fluoro-8-((1-(methyl-d3)-1H-pyrazol-4-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (90 mg, 0.23 mmol) and 2-aminopyrazolo[1,5-a]pyrimidine-3-carboxylic acid (41 mg, 0.23 mmol) in DCM (15 mL) were added [1,2,3]triazolo[4,5-b]pyridin-3-ol (47 mg, 0.34 mmol), EDCI (54 mg, 0.34 mmol) and DIEA (90 mg, 0.69 mmol), and the mixture was warmed to 40°C and stirred for 16 h. After cooling to room temperature, the mixture was diluted with DCM (50 mL) and washed with brine (30 mL x 2). The combined organic layers were dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by preparative HPLC (DCM / MeOH=10 / 1) to give (S)-2-amino-N-(1-(7-fluoro-8-((1-(methyl-d3)-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (48.5 mg, 38% yield) as a white solid. LCMS: MS (ESI) m / z 550.0 [M+H] + . 1 HNMR (400MHz, DMSO-d6) δ8.93(dd,J=6.7,1.6Hz,1H),8.55(dd,J=4.5,1.6Hz,1H),8.05(d,J=0.6Hz,1H),7.99(d,J=6.6Hz,1H),7.72-7.68(m,2H),7.64 -7.55(m,2H),7.55-7.45(m,3H),7.41-7.36(m,1H),7.02(dd,J=6.7,4.5Hz, 1H), 6.79 (s, 1H), 6.43 (s, 2H), 4.54 (q, J = 6.8Hz, 1H), 1.35 (d, J = 6.8Hz, 3H).

[0575] Example 29: 2-amino-N-((1S)-1-(8-(2-(1-methyl-5-oxopyrrolidin-3-yl)ethynyl)-1-oxo-2-phenylisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide; and Example 30: 6-(3-((S)-1-(2-aminopyrazolo[1,5-a]pyrimidine-3-carboxamido)ethyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-8-yl)-4-hydroxy-hex-5-ynoic acid methyl ester

[0576]

[0577] Step 1) 4-Hydroxy-6-(trimethylsilyl)hex-5-ynoic acid methyl ester

[0578] To a solution of methyl 4-oxo-6-(trimethylsilyl)hex-5-ynoate (300 mg, 1.4 mmol) in MeOH (10 mL) was added NaBH4 (59 mg, 1.55 mmol) at -20°C, and the mixture was stirred at room temperature for 16 h. The reaction was then quenched with NH4Cl solution (5 mL) and concentrated in vacuo. The residue was diluted with EtOAc (50 mL) and washed with brine (30 mL x 2). The separated organic layer was dried over Na2SO4 and concentrated in vacuo to give methyl 4-hydroxy-6-(trimethylsilyl)hex-5-ynoate (270 mg, yield 89%) as a colorless oil. MS (ESI): 215.2 [M+H] + .

[0579] Step 2) 4-Hydroxy-hex-5-ynoic acid methyl ester

[0580] To a solution of 4-hydroxy-6-(trimethylsilyl)hex-5-nonanoic acid methyl ester (250 mg, 1.17 mmol) in THF (10 mL) was added TBAF (1.2 mL, 1 M / L) at -10°C, and the mixture was stirred at room temperature for 1 h. The mixture was diluted with EtOAc (50 mL) and washed with brine (30 mL×2). The separated organic layer was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (EA:PE=1:4) to give 4-hydroxy-hex-5-ynoic acid methyl ester (125 mg, 60% yield) as a colorless oil. 1 H NMR (400MHz, CDCl3) δ4.50 (td, J = 6.1, 2.1Hz, 1H), 3.70 (s, 3H), 2.65-2.53 (m, 2H), 2.49 (d, J = 2.1Hz, 1H), 2.09-2.04 (m, 2H).

[0581] Step 3) 3-((1S)-1-aminoethyl)-8-(2-(5-oxooxolan-2-yl)ethynyl)-2-phenylisoquinolin-1-one; 6-(3-((S)-1-aminoethyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-8-yl)-4-hydroxy-hex-5-ynoic acid methyl ester

[0582] To a solution of 3-((1S)-1-aminoethyl)-8-chloro-2-phenylisoquinolin-1-one (60 mg, 0.2 mmol) and 4-hydroxy-hex-5-ynoic acid methyl ester (28 mg, 0.2 mmol) in CH3CN (10 mL) were added K3PO4 (128 mg, 0.6 mmol), X-phos (38 mg, 0.08 mmol) and Pd2(dba)3 (37 mg, 0.04 mmol), and the mixture was warmed to 80°C and stirred for 16 h. After cooling to room temperature, it was concentrated in vacuo. The residue was diluted with DCM (100 mL) and washed with brine (30 mL). The separated organic layer was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (MeOH / DCM=1 / 20) to give 3-((1S)-1-aminoethyl)-8-(2-(5-oxooxolan-2-yl)ethynyl)-2-phenylisoquinolin-1-one, MS (ESI): 373.1 [M+H] + ; and a mixture of 6-(3-((S)-1-aminoethyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-8-yl)-4-hydroxy-hex-5-ynoic acid methyl ester (60 mg, 60.16% yield) as a yellow solid, MS (ESI): 405.2 [M + H] + .

[0583] Step 4) 2-amino-N-((1S)-1-(8-(2-(1-methyl-5-oxopyrrolidin-3-yl)ethynyl)-1-oxo-2-phenylisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide; 6-(3-((S)-1-(2-aminopyrazolo[1,5-a]pyrimidine-3-carboxamido)ethyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-8-yl)-4-hydroxy-hex-5-ynoic acid methyl ester

[0584] To a mixture of 3-((1S)-1-aminoethyl)-8-(2-(5-oxooxolan-2-yl)ethynyl)-2-phenylisoquinolin-1-one and 6-(3-((S)-1-aminoethyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-8-yl)-4-hydroxy-hex-5-ynoic acid methyl ester (60 mg, 0.16 mmol) and 2-aminopyrazolo[1,5-a]pyrimidine-3-carboxylic acid (29 mg, 0.16 mmol) in DCM (10 mL) were added HOAT (33 mg, 0.24 mmol), EDCI (46 mg, 0.24 mmol) and DIEA (62 mg, 0.48 mmol), and the mixture was warmed to 40° C. and stirred for 16 h. The mixture was diluted with DCM (150 mL) and washed with brine (50 mL×2). The liquids were separated, and the organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by preparative HPLC (ACN-H2O 0.1% FA, gradient 30% to 60%) to give 2-amino-N-((1S)-1-(8-(2-(1-methyl-5-oxopyrrolidin-3-yl)ethynyl)-1-oxo-2-phenylisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (8.0 mg, 11.30% yield) as a white solid, and 6-(3-((S)-1-(2-aminopyrazolo[1,5-a]pyrimidine-3-carboxamido)ethyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-8-yl)-4-hydroxy-hex-5-ynoic acid methyl ester (3.4 mg, 3.74% yield) as a white solid.

[0585] 2-amino-N-((1S)-1-(8-(2-(1-methyl-5-oxopyrrolidin-3-yl)ethynyl)-1-oxo-2-phenylisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide: MS (ESI): 533.2 [M+H] + . 1 H NMR (400MHz, CD3OD) δ8.71(dd,J=6.8,1.6Hz,1H),8.54-8.51(m,1H),7.67-7.35(m,8H),6.98(dd,J=6.8,4.5Hz,1H),6 .90(d,J=4.1Hz,1H),5.47(t,J=6.4Hz,1H),4.80-4.74(m,1H),4.56(s,1H),2.66-2.41(m,4H),1.48(d,J=6.7Hz,3H).

[0586] 6-(3-((S)-1-(2-aminopyrazolo[1,5-a]pyrimidine-3-carboxamido)ethyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-8-yl)-4-hydroxy-hex-5-ynoic acid methyl ester: MS (ESI): 565.2 [M+H] + . 1 H NMR (400MHz, CD3OD) δ8.72(dd,J=6.8,1.6Hz,1H),8.53(dd,J=4.5,1.6Hz,1H),7.63-7.61(m,4H),7.54-7.47(m,4H),6.99(dd,J=6.8,4.5 Hz,1H),6.89(s,1H),4.83-4.77(m,1H),4.63-4.60(m,1H),3.64(s,3H),2.56(t,J=7.7Hz,2H),2.06–2.02(m,2H),1.48(d,J=6.8Hz,3H). Example 31 (S)-2-amino-N-(1-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-3-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0587]

[0588] Step 1) 4-bromo-1-(2-((tert-butyldimethylsilyl)oxy)ethyl)-1H-pyrazole

[0589] To a solution of 4-bromo-1H-pyrazole (2.0 g, 13.6 mmol) and (2-bromoethoxy)(tert-butyl)dimethylsilane (3.58 g, 149.6 mmol) in MeCN (100 mL) was added cesium carbonate (8.86 g, 27.2 mmol), and the mixture was stirred at room temperature for 16 h. The mixture was concentrated in vacuo, the residue was diluted with EtOAc (130 mL), washed with brine (50 mL x 2), the separated organic layer was dried over Na2SO4, and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=50 / 1) to give 4-bromo-1-(2-((tert-butyldimethylsilyl)oxy)ethyl)-1H-pyrazole (2.5 g, 70% yield) as a yellow oil. MS (ESI): 304.9 [M+H] + .

[0590] Step 2) 4-bromo-1-(2-((tert-butyldimethylsilyl)oxy)ethyl)-1H-pyrazole-5-carbaldehyde

[0591] To a solution of 4-bromo-1-(2-((tert-butyldimethylsilyl)oxy)ethyl)-1H-pyrazole (300 mg, 0.98 mmol) in THF (30 mL) was added LDA (0.6 mL, 1.18 mmol) at -78°C, and the mixture was stirred for 1 h. DMF (144 mg, 1.96 mmol) was then added, and the mixture was transferred to room temperature and stirred for 1 h. The mixture was quenched with aqueous NH4Cl (20 mL) solution and extracted with EtOAc (50 mL x 2), and washed with brine (40 mL x 2). The combined organic layers were dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=50 / 1) to give 4-bromo-1-(2-((tert-butyldimethylsilyl)oxy)ethyl)-1H-pyrazole-5-carbaldehyde (200 mg, 66% yield) as a yellow oil. MS(ESI):333.1[M+H] + .

[0592] Step 3) 3-Bromo-6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazine

[0593] To a solution of 4-bromo-1-(2-((tert-butyldimethylsilyl)oxy)ethyl)-1H-pyrazole-5-carbaldehyde (200 mg, 0.60 mmol) in DCM (20 mL) was added dropwise Et3SiH (209 mg, 1.80 mmol) and TMSOTf (800 mg, 3.60 mmol) under nitrogen atmosphere at -78°C, and the mixture was transferred to room temperature and stirred for 15 h. The mixture was quenched with aqueous NaHCO3 solution (20 mL) and extracted with DCM (50 mL x 3). The combined organic layers were washed with brine (50 mL x 2), dried over Na2SO4, and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=40 / 1) to give 3-bromo-6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazine (100 mg, 74% yield) as a yellow oil. MS(ESI):203.1[M+H] + .

[0594] Step 4) 3-((triisopropylsilyl)ethynyl)-6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazine

[0595] Under nitrogen atmosphere, X-Phos (117 mg, 0.25 mmol), Pd2(dba)3 (71 mg, 0.12 mmol) and K3PO4 (313 mg, 1.48 mmol) were added to a solution of 3-bromo-6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazine (100 mg, 0.49 mmol) in MeCN (20 mL), and the mixture was warmed to 80 ° C and stirred for 5 h. After cooling to room temperature, it was concentrated in vacuo. The residue was diluted with EtOAc (60 mL) and washed with brine (40 mL x 2). The separated organic layer was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=50 / 1) to give 3-((triisopropylsilyl)ethynyl)-6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazine (150 mg, 90% yield) as a yellow oil. MS (ESI): 305.2 [M+H] + .

[0596] Step 5) 3-ethynyl-6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazine

[0597] To a solution of 3-((triisopropylsilyl)ethynyl)-6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazine (150 mg, 0.49 mmol) in THF (10 mL) was added TBAF (1 mL, 1 mmol) at 0°C, and the mixture was stirred for 2 h. The mixture was diluted with H2O (30 mL) and extracted with EtOAc (30 mL x 3). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=30 / 1) to give 3-ethynyl-6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazine (114 mg, 78% yield) as a yellow oil. MS (ESI): 149.0 [M+H] + .

[0598] Step 6) (S)-3-(1-aminoethyl)-8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-3-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one

[0599] To a solution of (S)-3-(1-aminoethyl)-8-chloro-2-phenylisoquinolin-1(2H)-one (65 mg, 0.22 mmol) and 3-ethynyl-6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazine (113 mg, 0.76 mmol) in MeCN (20 mL) were added X-Phos (52 mg, 0.11 mmol), Pd2(dba)3 (31 mg, 0.05 mmol) and K3PO4 (138 mg, 0.65 mmol), and the mixture was warmed to 80°C and stirred for 5 h. After cooling to room temperature, the mixture was concentrated in vacuo. The residue was diluted with EtOAc (70 mL), washed with brine (50 mL x 2), dried over Na2SO4, and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=20 / 1) to give (S)-3-(1-aminoethyl)-8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-3-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (20 mg, 21% yield) as a yellow oil. MS (ESI): 411.1 [M+H] + .

[0600] Step 7) (S)-2-amino-N-(1-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-3-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0601] To a solution of (S)-3-(1-aminoethyl)-8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-3-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (20 mg, 0.05 mmol) and 2-amino-5-methylpyrazolo[1,5-a]pyrimidine-3-carboxylic acid (7 mg, 0.05 mmol) in DCM (10 mL) were added DIEA (17 mg, 0.15 mmol), EDCI (14 mg, 0.07 mmol) and HOAt (10 mg, 0.07 mmol), and the mixture was warmed to 40° C. and stirred for 16 h. The mixture was diluted with DCM (50 mL) and washed with brine (30 mL×2). The separated organic layer was dried over Na2SO4 and concentrated in vacuo. The residue was purified by preparative HPLC (-Gemini-C18 150 x 19 mm, 5um: ACN--H2O (0.05% NH3), 20-50) to give (S)-2-amino-N-(1-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-3-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (6.59 mg, 23% yield) as a yellow solid. MS (ESI): 570.8 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.92(dd,J=6.8,1.6Hz,1H),8.55(dd,J=4.4,1.6Hz,1H),8.00(d,J=6.8Hz,1H),7.67-7.55(m,5H),7.55-7.45(m,3H),7.41 -7.36(m,1H),7.01(dd,J=6.8,4.4Hz,1H),6.74(s,1H),6.42(s,2H),4.8 4(s,2H),4.56(q,J=6.4Hz,1H),4.13-4.02(m,4H),1.35(d,J=6.8Hz,3H).

[0602] Example 32 (S)-2-amino-N-(1-(5-((2,3-dihydropyrazolo[5,1-b]oxazol-6-yl)ethynyl)-6-fluoro-4-oxo-3-phenyl)-3,4-dihydroquinazolin-2-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0603]

[0604] Step 1) (S)-2-(1-aminoethyl)-5-((2,3-dihydropyrazolo[5,1-b]oxazol-6-yl)ethynyl)-6-fluoro-3-phenylquinazolin-4(3H)-one

[0605] Under nitrogen atmosphere, Pd2(dba)3 (32 mg, 0.034 mmol), X-phos (33 mg, 0.07 mmol) and K3PO4 (88 mg, 0.41 mmol) were added to a solution of 6-ethynyl-2,3-dihydropyrazolo[5,1-b]oxazole (50 mg, 0.14 mmol) and 2-ethynyl-5H,6H-pyrazolo[3,2-b][1,3]oxazole (18.5 mg, 0.14 mmol) in acetonitrile (20 mL), and the mixture was heated to 80°C and stirred for 4 h. After cooling to room temperature, the mixture was concentrated in vacuo. The residue was dissolved in DCM (50 mL), washed with water (30 ml) and brine (30 mL), and concentrated in vacuo. The residue was purified by column chromatography (DCM / MEOH=10 / 1) to give (S)-2-(1-aminoethyl)-5-((2,3-dihydropyrazolo[5,1-b]oxazol-6-yl)ethynyl)-6-fluoro-3-phenylquinazolin-4(3H)-one (20 mg, 30% yield) as a white solid. LCMS: MS (ESI) m / z 416.1 [M+H] + .

[0606] Step 2) (S)-2-amino-N-(1-(5-((2,3-dihydropyrazolo[5,1-b]oxazol-6-yl)ethynyl)-6-fluoro-4-oxo-3-phenyl)-3,4-dihydroquinazolin-2-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0607] To a solution of (S)-2-(1-aminoethyl)-5-((2,3-dihydropyrazolo[5,1-b]oxazol-6-yl)ethynyl)-6-fluoro-3-phenylquinazolin-4(3H)-one (20 mg, 0.048 mmol) and 2-aminopyrazolo[1,5-a]pyrimidine-3-carboxylic acid (8.6 mg, 0.048 mmol) in DCM (15 mL) were added HOAt (10 mg, 0.072 mmol), EDCI (11 mg, 0.072 mmol) and DIEA (18.6 mg, 0.14 mmol), and the mixture was warmed to 40°C and stirred for 16 h. After cooling to room temperature, the mixture was diluted with water (30 mL) and extracted with DCM (30 mL x 2). The combined organic layers were washed with brine (30 mL x 2), dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography (DCM / MEOH=10 / 1) to give (S)-2-amino-N-(1-(5-((2,3-dihydropyrazolo[5,1-b]oxazol-6-yl)ethynyl)-6-fluoro-4-oxo-3-phenyl)-3,4-dihydroquinazolin-2-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (4.14 mg, 14% yield) as a white solid. LCMS: MS (ESI) m / z 576.0 [M+H] + . 1 H NMR (400MHz, CDCl3) δ8.78(d,J=6.2Hz,1H),8.54(dd,J=4.4,1.7Hz,1H),8.42(dd,J=6.8,1.7Hz,1H),8.29(s,

[0608] 1H),7.72(d,J=7.4Hz,1H),7.68-7.63(m,1H),7.60-7.58(m,1H),7.56-7.51(m,1H),7.35-7.28(m,1H),6.81(dd ,J=6.8,4.4Hz,1H),5.65(s,1H),5.06-4.99(m,2H),4.88-4.82(m,1H),4.34-4.27(m,2H),1.71(d,J=6.9Hz,3H).

[0609] Example 33 2-amino-N-((S)-1-(8-(((R)-5-hydroxy-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0610]

[0611] Step 1) (2S,4R)-4-(Benzyloxy)pyrrolidine-2-carboxylic acid

[0612] To a solution of (2S,4R)-4-(benzyloxy)-1-(tert-butoxycarbonyl)pyrrolidine-2-carboxylic acid (4 g, 12.4 mmol) in DCM (20 mL) was added TFA (4 mL) and the mixture was stirred for 2 h. The mixture was concentrated in vacuo to give (2S,4R)-4-(benzyloxy)pyrrolidine-2-carboxylic acid (3 g, 98.4% yield) as a yellow oil. MS (ESI): 222.1 [M+H] + .

[0613] Step 2) (2S,4R)-4-(Benzyloxy)-1-nitrosopyrrolidine-2-carboxylic acid

[0614] To a solution of (2S,4R)-4-(benzyloxy)pyrrolidine-2-carboxylic acid (3 g, 13.6 mmol) in H2O (10 mL) was added a solution of NaNO2 (1.41 g, 20.4 mmol) in H2O (5 mL) at 0°C, followed by HOAc (3 mL), and the mixture was stirred at room temperature for 16 h. Filter, wash the filter cake with water (10 mL), and dry in vacuo to give the product (3.5 g, 98% yield) as a white solid. MS (ESI): 251.1 [M+H] + .

[0615] Step 3) (R)-5-(Benzyloxy)-3-oxo-5,6-dihydro-3H-pyrrolo[1,2-c][1,2,3]oxadiazole-7(4H)-ammonium-3a-ylide

[0616] To a solution of (2S,4R)-4-(benzyloxy)-1-nitrosopyrrolidine-2-carboxylic acid (3.5 g, 14 mmol) in THF (50 mL) was added TFAA (4.41 g, 21 mmol) at 0°C, and the mixture was stirred at room temperature for 16 h. The mixture was diluted with EtOAc (80 mL) and washed with brine (50 mL×2). The separated organic layer was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (EA / PE=1 / 2) to give (R)-5-(benzyloxy)-3-oxo-5,6-dihydro-3H-pyrrolo[1,2-c][1,2,3]oxadiazole-7(4H)-ammonium-3a-ylide (1.5 g, 41.4% yield) as a yellow oil. MS (ESI): 233.1 [M+H] + .

[0617] Step 4) (R)-5-(Benzyloxy)-2-(trimethylsilyl)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole

[0618] A mixed solution of (R)-5-(benzyloxy)-3-oxo-5,6-dihydro-3H-pyrrolo[1,2-c][1,2,3]oxadiazole-7(4H)-ammonium-3a-ylide (1 g, 4.3 mmol) and ethynyltrimethylsilane (0.84 g, 8.6 mmol) in xylene (10 mL) was heated to 130°C for 16 h. After cooling to room temperature, it was concentrated in vacuo. The residue was purified by column chromatography (EA / PE=1 / 3) to give (R)-5-(benzyloxy)-2-(trimethylsilyl)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole (600 mg, 46.5% yield) as a yellow oil. MS (ESI): 287.1 [M+H] + .

[0619] Step 5) (R)-5-(Benzyloxy)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole

[0620] To a solution of (R)-5-(benzyloxy)-2-(trimethylsilyl)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole (700 mg, 2.44 mmol) in THF (5 mL) was added TBAF (24.4 mL, 24.4 mmol), and the mixture was heated to 60°C and heated for 48 h. After cooling to room temperature, the mixture was diluted with EtOAc (40 mL) and washed with brine (30 mL×2). The separated organic layer was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (EA / PE=1 / 2) to give (R)-5-(benzyloxy)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole (330 mg, 56.7% yield) as a yellow solid. MS (ESI): 215.1 [M+H] + .

[0621] Step 6) (R)-5-(Benzyloxy)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole-3-carbaldehyde

[0622] To a solution of (R)-5-(benzyloxy)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole (330 mg, 1.54 mmol) in DCM (20 mL) was added (chloromethylene)dimethylammonium chloride (296 mg, 2.31 mmol), and the mixture was warmed to 40 °C and heated for 48 h. 1N HCl (3 mL) was added to the mixture, and the temperature was raised to 40 °C and heated for 2 h. It was then concentrated in vacuo, and the residue was diluted with water (5 mL) and washed with aqueous NaHCO3 solution (10 mL). The mixture was extracted with EtOAc (30 mL x 3), and the combined organic layers were washed with brine (20 mL), dried over Na2SO4, and concentrated in vacuo to give (R)-5-(benzyloxy)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole-3-carbaldehyde (350 mg, yield 84%) as a yellow oil. MS(ESI):243.1[M+H] + Step 7) (R)-5-Hydroxy-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole-3-carbaldehyde

[0623] (R)-5-(Benzyloxy)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole-3-carboxaldehyde (200 mg, 0.82 mmol) was dissolved in concentrated HCl (10 mL), heated to 70 °C and stirred for 16 h. After cooling to room temperature, it was concentrated in vacuo. The residue was diluted with water (3 mL), washed with aqueous sodium bicarbonate solution (10 mL), and then extracted with EtOAc (20 mL x 3). The combined organic layers were dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (EA / PE=1 / 3) to give (R)-5-hydroxy-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole-3-carboxaldehyde (140 mg, 89% yield) as a yellow solid. MS (ESI): 153.1 [M+H] + . 1 H NMR(400MHz, CDCl3)δ9.82(s,1H),7.97(s,1H),5.21-5.17(m,1H),4.40(dd,J=12.4,6.0Hz,1 H), 4.16 (dd, J = 12.4, 2.4Hz, 1H), 3.46 (dd, J = 17.6, 6.4Hz, 1H), 3.15 (dd, J = 17.6, 2.4Hz, 1H).

[0624] Step 8) (R)-3-ethynyl-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-5-ol

[0625] K2CO3 (91 mg, 0.66 mmol) was added to a solution of (R)-5-hydroxy-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole-3-carbaldehyde (50 mg, 0.33 mmol) and dimethyl (1-diazo-2-oxopropyl)phosphonate (95 mg, 0.49 mmol) in MeOH (20 mL), and the mixture was stirred at room temperature for 16 h. The mixture was concentrated in vacuo, diluted with EtOAc (60 mL), and washed with brine (20 mL×2). The separated organic layer was dried over Na2SO4 and concentrated in vacuo to give (R)-3-ethynyl-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole-5-ol (45 mg, 83% yield) as a yellow oil. MS (ESI): 148.9 [M+H] + . 1 HNMR (400MHz, CDCl3) δ7.62 (s, 1H), 5.15-5.07 (m, 1H), 4.35 (dd, J = 12.0, 6.0 Hz, 1H), 4.09 (dd, J = 12.0, 2. 8Hz, 1H), 3.77 (d, J = 10.8Hz, 1H), 3.30 (dd, J = 16.8, 6.4Hz, 1H), 3.02 (s, 1H), 2.93 (dd, J = 16.8, 2.8Hz, 1H).

[0626] Step 9) 3-((S)-1-aminoethyl)-8-(((R)-5-hydroxy-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one

[0627] Under nitrogen atmosphere, to a solution of 3-((1S)-1-aminoethyl)-8-chloro-2-phenylisoquinolin-1-one (50 mg, 0.17 mmol) and 4-ethynyl-1-methylpyrazole (37 mg, 0.25 mmol) in CH3CN (20 mL) were added Pd2(dba)3 (31 mg, 0.033 mmol), K3PO4 (71 mg, 0.33 mmol) and X-phos (32 mg, 0.067 mmol), and the mixture was heated to 80°C and stirred for 5 h. After cooling to room temperature, the mixture was diluted with EtOAc (60 mL) and washed with brine (40 mL×2). The separated organic layer was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=20 / 1) to give 3-((S)-1-aminoethyl)-8-(((R)-5-hydroxy-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (40 mg, 61%) as a yellow solid. MS (ESI): 394.0 [M+H] + .

[0628] Step 10) 2-amino-N-((S)-1-(8-(((R)-5-hydroxy-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0629] To a solution of 3-((S)-1-aminoethyl)-8-(((R)-5-hydroxy-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (40 mg, 0.1 mmol) and 2-aminopyrazolo[1,5-a]pyrimidine-3-carboxylic acid (19.5 mg, 0.1 mmol) in DCM (20 mL) were added EDCI (31.5 mg, 0.16 mmol), HOAt (18 mg, 0.13 mmol) and DIEA (43 mg, 0.32 mmol), and the mixture was warmed to 40°C and stirred for 16 h. After cooling to room temperature, the mixture was diluted with DCM (40 mL) and washed with brine (20 mL x 2). The separated organic layer was dried over Na2SO4 and concentrated in vacuo. The residue was purified by preparative HPLC (ACN-H2O, 0.1% FA, gradient 20% to 50%) to give 2-amino-N-((S)-1-(8-(((R)-5-hydroxy-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (14.6 mg, 23% yield) as a light yellow solid. MS (ESI): 571.2 [M+H] + . 1 H NMR (400MHz, CD3OD) δ8.62(dd,J=6.8,1.6Hz,1H),8.43(dd,J=4.4,1.6Hz,1H),8.14(d,J=6.8Hz, 1H),7.61(s,1H),7.57-7.54(m,2H),7.52-7.48(m,2H),7.45-7.33(m,4H),6.89(dd,J=6.8,4.4H z,1H),6.80(s,1H),4.93-4.90(m,1H),4.72-4.65(m,1H),4.24(dd,J=11.6,5.6Hz,1H),3.88(dd ,J=11.6,2.4Hz,1H),3.19(d,J=6.4Hz,1H),2.82(dd,J=16.8,2.4Hz,1H),1.39(d,J=6.8Hz,3H).

[0630] Example 34 (S)-2-amino-N-(1-(7-fluoro-8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-(phenyl-d5)-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0631]

[0632] Step 1) 2-Chloro-3-fluoro-6-methyl-N-(phenyl-d5)benzamide

[0633] Oxalyl chloride (810 mg, 6.36 mmol) was added dropwise to a solution of 2-chloro-3-fluoro-6-methylbenzoic acid (1.0 g, 5.3 mmol) and DMF (40 mg, 0.53 mmol) in DCM (25 mL) at 0°C. The mixture was stirred for 2 h after the addition. The mixture was concentrated under reduced pressure to obtain a mixture of acyl chlorides. A solution of the above acyl chlorides in DCM (10 mL) was added to a solution of benzene-d5-amine (550 mg, 5.3 mmol) and TEA (1.61 g, 15.9 mol) in DCM (25 mL) at 0°C. The mixture was warmed to room temperature and stirred for 5 h. The mixture was diluted with H2O (30 mL) and extracted with DCM (30 mL x 3). The combined organic layers were washed with brine (50 mL x 2), dried over Na2SO4, and concentrated under reduced pressure. The residue was purified by column chromatography (PE / EA=3 / 1) to give 2-chloro-3-fluoro-6-methyl-N-(phenyl-d5)benzamide (1.01 g, 64% yield) as a yellow solid. MS (ESI): 269.1 [M+H] + .

[0634] Step 2) Tert-butyl (S)-(4-(3-chloro-4-fluoro-2-((phenyl-d5)carbamoyl)phenyl)-3-oxobutan-2-yl)carbamate

[0635] Under nitrogen atmosphere, a solution of 2-chloro-3-fluoro-6-methyl-N-(phenyl-d5)benzamide (1.01 g, 3.8 mmol) in THF (25 mL) was cooled to -30 °C, n-BuLi (4.0 mL, 9.5 mmol) was added, and stirred for 1 h to obtain a yellow liquid. At -30 °C, a solution of (S)-(1-(methoxy(methyl)amino)-1-oxopropane-2-yl)carbamic acid tert-butyl ester (1.32 g, 5.7 mmol) in THF (25 mL) was added and stirred for 1 h. The mixture was then slowly added to the above yellow mixture, transferred to room temperature, and stirred for 3 hours. The mixture was diluted with NH4Cl (30 mL) aqueous solution and extracted with EtOAc (50 mL x 3). The combined organic layer was washed with brine (40 mL x 2), dried over Na2SO4, and concentrated under reduced pressure. The residue was purified by column chromatography (PE / EA=3 / 1) to give (S)-tert-butyl(4-(3-chloro-4-fluoro-2-((phenyl-d5)carbamoyl)phenyl)-3-oxobutan-2-yl)carbamate (0.63 g, 37% yield) as a yellow solid. MS (ESI): 462.1 [M+Na] + .

[0636] Step 3) (S)-3-(1-aminoethyl)-8-chloro-7-fluoro-2-(phenyl-d5)isoquinolin-1(2H)-one

[0637] To a solution of (S)-tert-butyl(4-(3-chloro-4-fluoro-2-((phenyl-d5)carbamoyl)phenyl)-3-oxobutan-2-yl)carbamate (0.63 g, 1.43 mmol) in MeOH (30 mL) was added concentrated HCl (15 mL), and the mixture was warmed to 80 ° C and stirred for 24 h. After cooling to room temperature, the mixture was concentrated under reduced pressure. The residue was diluted with H2O (10 mL) and the pH was adjusted to 7-8 with aqueous NaHCO3. The mixture was extracted with DCM (60 mL x 3) and washed with brine (50 mL x 2). The separated organic layer was dried over Na2SO4 and concentrated under reduced pressure. The residue was purified by column chromatography (DCM / MeOH=95 / 5) to give (S)-3-(1-aminoethyl)-8-chloro-7-fluoro-2-(phenyl-d5)isoquinolin-1(2H)-one (160 mg, 33% yield) as a yellow solid. MS (ESI): 321.9 [M+H] + .

[0638] Step 4) (S)-3-(1-aminoethyl)-7-fluoro-8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2-(phenyl-d5)isoquinolin-1(2H)-one

[0639] To a solution of (S)-3-(1-aminoethyl)-8-chloro-7-fluoro-2-(phenyl-d5)isoquinolin-1(2H)-one (160 mg, 0.50 mmol) and 4-ethynyl-1-methyl-1H-pyrazole (79 mg, 0.75 mmol) in MeCN (10 mL) were added X-Phos (118 mg, 0.25 mmol), Pd2(dba)3 (72 mg, 0.12 mmol) and K3PO4 (317 mg, 1.50 mmol), and the mixture was warmed to 80°C and stirred for 5 h. After cooling to room temperature, it was concentrated in vacuo. The residue was diluted with EtOAc (80 mL) and washed with brine (50 mL x 2). The separated organic layer was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=20 / 1) to give (S)-3-(1-aminoethyl)-7-fluoro-8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2-(phenyl-d5)isoquinolin-1(2H)-one (110 mg, 53% yield) as a yellow oil. MS (ESI): 391.9 [M+H] + .

[0640] Step 5) (S)-2-amino-N-(1-(7-fluoro-8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-(phenyl-d5)-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0641] To a solution of (S)-3-(1-aminoethyl)-7-fluoro-8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2-(phenyl-d5)isoquinolin-1(2H)-one (110 mg, 0.28 mmol) and 2-amino-5-methylpyrazolo[1,5-a]pyrimidine-3-carboxylic acid (55 mg, 0.31 mmol) in DCM (30 mL) were added DIEA (109 mg, 0.84 mmol), EDCI (81 mg, 0.42 mmol) and HOAt (57 mg, 0.42 mmol), and the mixture was warmed to 40° C. and stirred for 16 h. After cooling to room temperature, the mixture was diluted with DCM (60 mL) and washed with brine (40 mL×2). The separated organic layer was dried over Na2SO4 and concentrated in vacuo. The residue was purified by preparative HPLC (-Gemini-C18 150x 19mm, 5um: ACN--H2O (0.05% NH3), 20-50) to give (S)-2-amino-N-(1-(7-fluoro-8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-(phenyl-d5)-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (31.8 mg, 20% yield) as a yellow solid. MS (ESI): 552.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.93(dd,J=6.8,1.6Hz,1H),8.55(dd,J=4.4,1.6Hz,1H),8.05(s,1H),8.00(d,J=6.8Hz,1H),7.70(ddd,J=21.2,13 .2,7.2Hz,2H),7.61(s,1H),7.02(dd,J=6.8,4.4Hz,1H),6.79(s,1H),6.43(s,2H),4.58-4.50(m,1H),3.82(s,3H),1.34(d,J=6.8Hz,3H).

[0642] Example 35 (S)-2-amino-N-(1-(7-fluoro-8-((1-methyl-1H-pyrazol-4-yl-5-d)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0643]

[0644] Step 1) 1-methyl-1H-pyrazole-5-d

[0645] To a solution of 5-bromo-1-methyl-1H-pyrazole (2 g, 12.42 mmol) in THF (10 mL) was added i-PrMgCl.LiCl (2.16 mg, 14.9 mmol; 1.3 M in THF) at -15°C. The mixture was warmed to -10-0°C and stirred for 1 h. Methanol-d4 (0.5 mL) was added to the reaction mixture at -10°C and stirred at room temperature for 16 h. The mixture was quenched with aqueous NH4Cl solution (5 mL) and MTBE (50 mL); the layers were separated and the organic phase was washed with brine (20 mL x 2). The separated organic layer was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to give 1-methyl-1H-pyrazole-5-d (500 mg, 48% yield) as a colorless oil. LCMS: MS (ESI) m / z 84.1 [M+H] + .

[0646] Step 2) 4-iodo-1-methyl-1H-pyrazole-5-d

[0647] Elemental iodine (763 mg, 3 mmol) was added to a solution of 1-methyl-1H-pyrazole-5-d (500 mg, 6 mmol) in water (10 mL), and the mixture was stirred at room temperature for 0.5 h, then H2O2 (3 mL) was added, the temperature was raised to 60°C, and stirred for 1 h. After cooling to room temperature, the reaction was quenched with aqueous sodium sulfite solution (30 mL). The mixture was extracted with EtOAc (50 mL x 2), and the combined organic layers were washed with brine (50 mL x 2), dried over anhydrous Na2SO4, and concentrated in vacuo. The residue was purified by column chromatography (PE:EA=3:1) to give 4-iodo-1-methyl-1H-pyrazole-5-d (1 g, 79% yield) as a yellow solid. LCMS:MS (ESI) m / z 210.1 [M+H] + .

[0648] Step 3) 1-methyl-4-((trimethylsilyl)ethynyl)-1H-pyrazole-5-d

[0649] To a solution of 4-iodo-1-methyl-1H-pyrazole-5-d (1 g, 4.78 mmol) and ethynyltrimethylsilane (705 mg, 7.18 mmol) in THF (100 mL) were added CuI (91 mg, 0.48 mmol), Pd(PPh3)2Cl2 (168 mg, 0.24 mmol) and triethylamine (968 mg, 9.57 mmol) at 0°C, and the mixture was stirred at room temperature for 2 hours. The residue was diluted with EtOAc (300 mL) and washed with brine (200 mL x 2). The separated organic layer was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography (PE:EA=3:1) to give 1-methyl-4-((trimethylsilyl)ethynyl)-1H-pyrazole-5-d (700 mg, 81% yield) as a yellow oil. LCMS:MS(ESI)m / z 180.2[M+H] + .

[0650] Step 4) 4-ethynyl-1-methyl-1H-pyrazole-5-d

[0651] To a solution of 1-methyl-4-((trimethylsilyl)ethynyl)-1H-pyrazole-5-d (700 mg, 3.9 mmol) in THF (20 mL) was added TBAF (4 mL, 4 mmol, 1 M in THF), and the mixture was stirred at room temperature for 1 h. The mixture was then diluted with EtOAc (50 mL) and washed with brine (30 mL x 2). The separated organic layer was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (PE:EA=3:1) to give 4-ethynyl-1-methyl-1H-pyrazole-5-d (200 mg, 47% yield) as a yellow oil. LCMS:MS (ESI) m / z 108.0 [M+H] + .

[0652] Step 5) (S)-3-(1-aminoethyl)-7-fluoro-8-((1-methyl-1H-pyrazol-4-yl-5-d)ethynyl)-2-phenylisoquinolin-1(2H)-one

[0653] Pd2(dba)3 (29 mg, 0.032 mmol), X-phos (30 mg, 0.063 mmol) and K3PO4 (67 mg, 0.32 mmol) were added to a solution of (S)-3-(1-aminoethyl)-8-chloro-7-fluoro-2-phenylisoquinolin-1(2H)-one (50 mg, 0.16 mmol) and 4-ethynyl-1-methyl-1H-pyrazole-5-d (25 mg, 0.24 mmol) in MeCN (20 mL), and the mixture was warmed to 80°C and stirred for 4 h. After cooling to room temperature, it was concentrated in vacuo. The residue was diluted with DCM (50 mL) and washed with brine (30 mL x 2). The separated organic layer was dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography (DCM:MeOH=10:1) to give (S)-3-(1-aminoethyl)-7-fluoro-8-((1-methyl-1H-pyrazol-4-yl-5-d)ethynyl)-2-phenylisoquinolin-1(2H)-one (20 mg, 32% yield) as a yellow solid. LCMS: MS (ESI) m / z 388.1 [M+H] + .

[0654] Step 6) (S)-2-amino-N-(1-(7-fluoro-8-((1-methyl-1H-pyrazol-4-yl-5-d)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0655] To a solution of (S)-3-(1-aminoethyl)-7-fluoro-8-((1-methyl-1H-pyrazol-4-yl-5-d)ethynyl)-2-phenylisoquinolin-1(2H)-one (20 mg, 0.052 mmol) and 2-aminopyrazolo[1,5-a]pyrimidine-3-carboxylic acid (9 mg, 0.052 mmol) in DCM (15 mL) were added HOAt (10 mg, 0.078 mmol), EDCI (12 mg, 0.078 mmol) and DIEA (20 mg, 0.155 mmol), and the mixture was warmed to 40°C and stirred for 16 h. After cooling to room temperature, the mixture was diluted with DCM (50 mL) and washed with brine (30 mL x 2). The separated organic layer was dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography (DCM:MeOH=10:1) to give (S)-2-amino-N-(1-(7-fluoro-8-((1-methyl-1H-pyrazol-4-yl-5-d)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (7.3 mg, 25% yield) as a white solid. LCMS: MS (ESI) m / z 548.0 [M+H]+ . 1 H NMR(400MHz,CD3OD)δ8.72(dd,J=6.8,1.6Hz,1H),8.53(dd,J=4.4,1.6Hz,1H),7.69(s,1H),7.67-7.60(m,2H),7.5 9-7.42(m,5H),6.99(dd,J=6.8,4.4Hz,1H),6.91(s,1H),4.77(q,J=6.8Hz,1H),3.89(s,3H),1.48(d,J=6.8Hz,3H).

[0656] Example 36 (S)-2-amino-N-(1-(5-fluoro-8-((1-(methyl-d3)-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0657]

[0658] Step 1) 6-Chloro-2-methyl-3-nitrobenzoic acid

[0659] At 0°C, nitric acid (4.1 g, 0.064 mol, 2.6 mL) was added dropwise to a solution of 2-chloro-6-methylbenzoic acid (10 g, 0.0586 mol) in sulfuric acid (100 mL). The mixture was stirred for 30 minutes, then transferred to room temperature and stirred for 16 h. The mixture was slowly poured into ice water (260 mL) and continued to stir for 1 h. Filter, wash the filter cake with water (50 mL), and vacuum dry to obtain 6-chloro-2-methyl-3-nitrobenzoic acid (12.5 g, yield 98%) as a white solid. LCMS: MS (ESI) m / z 214.4 [MH] - .

[0660] Step 2) 6-Chloro-2-methyl-3-nitro-N-phenylbenzamide

[0661] At 0°C, DMF (1 mL) and oxalyl chloride (8.2 g, 0.064 mol) were added to a solution of 6-chloro-2-methyl-3-nitrobenzoic acid (11.5 g, 0.053 mol) in DCM (60 mL), and the mixture was transferred to room temperature and stirred for 2 h; the mixture was concentrated in vacuo to give a white solid (13 g). At 0°C, the above white solid (13 g) was dissolved in DCM (20 mL), aniline (7.5 g, 0.08 mol), triethylamine (5.4 g, 0.0533 mol) and DCM (60 mL) were added, and the mixture was stirred at room temperature for 16 hours. The mixture was quenched with water (200 mL) and extracted with DCM (200 mL x 2). The combined organic layers were washed with brine (200 mL x 2), dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to give 6-chloro-2-methyl-3-nitro-N-phenylbenzamide (14 g, 89% yield) as a yellow solid. LCMS: MS (ESI) m / z 291.0 [M+H] + .

[0662] Step 3) 3-amino-6-chloro-2-methyl-N-phenylbenzamide

[0663] SnCl2 (29.7 g, 0.155 mol) was added to a solution of 6-chloro-2-methyl-3-nitro-N-phenylbenzamide (15 g, 0.051 mol) in MeOH (200 mL), and the mixture was warmed to reflux and stirred for 24 h. After cooling to room temperature, it was concentrated in vacuo. The residue was quenched with aqueous NaHCO3 (200 mL) and extracted with EtOAc (300 mL x 2). The combined organic layers were washed with brine (200 mL x 2), dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography (PE / EA=3 / 1) to give 3-amino-6-chloro-2-methyl-N-phenylbenzamide (7 g, 51% yield) as a white solid. LCMS:MS (ESI) m / z 261.1 [M+H] + .

[0664] Step 4) 6-Chloro-3-fluoro-2-methyl-N-phenylbenzamide

[0665] At 0°C, slowly add sodium nitrate (1g, 15.34mmol) aqueous solution (5mL) to a solution of 3-amino-6-chloro-2-methyl-N-phenylbenzamide (2g, 7.67mmol) in tetrafluoroboric acid (48mass%, 20mL). After the addition is complete, the mixture is transferred to room temperature and stirred for 1h. Filter, rinse the filter cake with cold water (5mL x 2), and dry in vacuo to obtain a diazonium salt (2g) as a white solid. Heat the diazonium salt at 120°C for 30 minutes, cool to room temperature, pour into ice water (50mL), and then extract with ethyl acetate (50mL×3). The combined organic layer is washed with brine (50mL x 2), dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography (PE / EA=10 / 1) to give 6-chloro-3-fluoro-2-methyl-N-phenylbenzamide (800 mg, yield 35%) as a white solid. LCMS: MS (ESI) m / z 264.1 [M+H] + .

[0666] Step 5) Tert-butyl (S)-(4-(3-chloro-6-fluoro-2-(phenylcarbamoyl)phenyl)-3-oxobutan-2-yl)carbamate

[0667] To a solution of 6-chloro-3-fluoro-2-methyl-N-phenylbenzamide (400 mg, 1.52 mmol) in THF (5 mL) at -30°C, n-BuLi (3.8 mmol, 2.4 M / L = 1.58 mL) was added, and the mixture was stirred for 2 h to give a yellow liquid. To a solution of tert-butyl N-((1S)-1-(methoxy(methyl)carbamoyl)ethyl)carbamate (528 mg, 2.27 mmol) in THF (10 mL) was added i-PrMgCl.LiCl (4.55 mmol, 1.3 M / L, 3.5 mL) at -30°C under a nitrogen atmosphere, and the mixture was stirred for 2 h. The above yellow liquid was added to the mixture at -40°C, transferred to room temperature, and stirred for 16 h. The mixture was quenched with NH4Cl solution (50 mL) and extracted with EtOAc (50 mL × 2). The combined organic layers were washed with brine (50 mL x 2), dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography (PE / EA=3 / 1) to give (S)-tert-butyl (4-(3-chloro-6-fluoro-2-(phenylcarbamoyl)phenyl)-3-oxobutan-2-yl)carbamate (160 mg, 24% yield) as a yellow solid. LCMS: MS (ESI) m / z 457.0 [M+Na] + .

[0668] Step 6) (S)-3-(1-aminoethyl)-8-chloro-5-fluoro-2-phenylisoquinolin-1(2H)-one

[0669] To a solution of (S)-tert-butyl (4-(3-chloro-6-fluoro-2-(phenylcarbamoyl)phenyl)-3-oxobutan-2-yl)carbamate (150 mg, 0.345 mmol) in MeOH (6 mL) was added concentrated hydrochloric acid (3 mL), and the mixture was warmed to 70°C and stirred for 3 h. After cooling to room temperature, it was concentrated in vacuo. The residue was poured into water (50 mL), and the resulting solution was neutralized with aqueous sodium bicarbonate to pH = 8. The mixture was extracted with EtOAc (50 mL x 3), and the combined organic layers were washed with brine (50 mL), dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=95 / 5) to give (S)-3-(1-aminoethyl)-8-chloro-5-fluoro-2-phenylisoquinolin-1(2H)-one (50 mg, 45% yield) as a white solid. LCMS:MS(ESI)m / z 317.0[M+H] + .

[0670] Step 7) (S)-3-(1-aminoethyl)-5-fluoro-8-((1-(methyl-d3)-1H-pyrazol-4-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one

[0671] Under nitrogen atmosphere, to a solution of (S)-3-(1-aminoethyl)-8-chloro-5-fluoro-2-phenylisoquinolin-1(2H)-one (50 mg, 0.23 mmol) and 4-ethynyl-1-methylpyrazole (26 mg, 0.23 mmol) in acetonitrile (20 mL) were added Pa2(dba)3 (36 mg, 0.039 mmol), X-phos (38 mg, 0.039 mmol) and K3PO4 (67 mg, 0.32 mmol), and the mixture was warmed to 80°C and stirred for 4 h. After cooling to room temperature, it was concentrated in vacuo. The residue was dissolved in DCM (50 mL) and washed with brine (30 mL). The separated organic layer was dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=10 / 1) to give (S)-3-(1-aminoethyl)-5-fluoro-8-((1-(methyl-d3)-1H-pyrazol-4-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (50 mg, 80% yield) as a yellow solid. LCMS: MS (ESI) m / z 390.1 [M+H] + .

[0672] Step 8) (S)-2-amino-N-(1-(5-fluoro-8-((1-(methyl-d3)-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0673] To a solution of (S)-3-(1-aminoethyl)-5-fluoro-8-((1-(methyl-d3)-1H-pyrazol-4-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (50 mg, 0.13 mmol) and 2-aminopyrazolo[1,5-a]pyrimidine-3-carboxylic acid (23 mg, 0.13 mmol) in DCM (15 mL) were added HOAt (26 mg, 0.19 mmol), EDCI (30 mg, 0.19 mmol) and DIEA (50 mg, 0.39 mmol), and the mixture was warmed to 40°C and stirred for 16 h. After cooling to room temperature, the mixture was diluted with DCM (50 mL) and washed with brine (30 mL x 2). The separated organic layer was dried over anhydrous Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (DCM:MeOH=15:1) to give (S)-2-amino-N-(1-(5-fluoro-8-((1-(methyl-d3)-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (24.5 mg, 34% yield) as a white solid. LCMS: MS (ESI) m / z 550.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.94(dd,J=6.8,1.6Hz,1H),8.56(dd,J=4.4,1.6Hz,1H),8.08(d,J=6.8Hz,1H),8.00(d,J=0.8Hz,1H),7.62-7 .49(m,7H),7.43(d,J=7.2Hz,1H),7.03(dd,J=6.8,4.4Hz,1H),6.75(s,1H),6.45(s,2H),4.55(t,J=6.8Hz,1H),1.36(d,J=6.8Hz,3H).

[0674] Example 37 (S) -2-amino-5-(methoxy-d3)-N-(1-(8-((1-(methyl-d3)-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0675]

[0676] Step 1) (S)-3-(1-aminoethyl)-8-((1-(methyl-d3)-1H-pyrazol-4-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one

[0677] Pd2(dba)3 (61 mg, 0.067 mmol), K3PO4 (142 mg, 0.67 mmol) and X-phos (64 mg, 0.13 mmol) were added to a solution of 3-((1S)-1-aminoethyl)-8-chloro-2-phenylisoquinolin-1-one (100 mg, 0.33 mmol) and 4-ethynyl-1-methylpyrazole (55 mg, 0.5 mmol) in CH3CN (20 mL), and the mixture was heated to 80°C and stirred for 5 hours. After cooling to room temperature, the mixture was diluted with EtOAc (60 mL) and washed with brine (40 mL×2). The separated organic layer was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=20 / 1) to give (S)-3-(1-aminoethyl)-8-((1-(methyl-d3)-1H-pyrazol-4-yl))ethynyl)-2-phenylisoquinolin-1(2H)-one (80 mg, 61%) as a yellow solid. MS (ESI): 371.9 [M+H] + .

[0678] Step 2) (S)-2-amino-5-(methoxy-d3)-N-(1-(8-((1-(methyl-d3)-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0679] To a solution of (S)-3-(1-aminoethyl)-8-((1-(methyl-d3)-1H-pyrazol-4-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (40 mg, 0.1 mmol) and 2-amino-5-methoxypyrazolo[1,5-a]pyrimidine-3-carboxylic acid (23 mg, 0.1 mmol) in DCM (20 mL) were added EDCI (31 mg, 0.16 mmol), HOAt (18 mg, 0.13 mmol) and DIEA (42 mg, 0.32 mmol), and the mixture was warmed to 40°C and stirred for 16 h. After cooling to room temperature, the mixture was diluted with DCM (40 mL) and washed with brine (30 mL x 2). The separated organic layer was dried over Na2SO4 and concentrated in vacuo. The residue was purified by preparative HPLC (DCM / MeOH=15 / 1) to give (S)-2-amino-5-(methoxy-d3)-N-(1-(8-((1-(methyl-d3)-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (19 mg, 30% yield) as a white solid. MS (ESI): 564.8 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.72-8.66(m,1H),8.01(d,J=0.8Hz,1H),7.88(d,J=6.8Hz,1H),7.68-7.61(m,2H),7.60(s,1H),7.59-7.50(m,4 H),7.46(d,J=7.6Hz,1H),7.39(d,J=7.6Hz,1H),6.79(s,1H),6.49-6.44(m,1H),6.14(s,2H),4.49-4.43(m,1H),1.36(d,J=6.8Hz,3H).

[0680] Example 38 (S)-2-amino-5-cyano-N-(1-(8-((1-(methyl-d3)-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyridine-3-carboxamide

[0681]

[0682] Step 1) 1-amino-4-bromopyridin-1-ium

[0683] To a solution of 4-bromopyridine (1 g, 6.3 mmol) in DCM (10 mL) was added aminodiphenylphosphinate (4.41 g, 18.9 mmol) in portions, and the mixture was stirred at room temperature for 48 h. Hydroiodic acid (1 mL, 55% in H2O) was then added, the precipitate was filtered, the filter cake was washed with DCM (10 mL), and vacuum dried to give 1-amino-4-bromopyridin-1-ium (1 g, 46% yield) as a brown solid. MS (ESI): 173.0, 175.0 [M+H] + .

[0684] Step 2) 5-bromopyrazolo[1,5-a]pyridine-2,3-dicarboxylic acid dimethyl ester

[0685] 1,4-Dimethylsuccinic acid (784 mg, 5.5 mmol) and K2CO3 (953 mg, 6.89 mmol) were added to a solution of 1-amino-4-bromopyridin-1-ium (800 mg, 4.6 mmol) in DMF (20 mL), and the mixture was stirred at room temperature for 16 h. The mixture was diluted with EtOAc (80 mL) and washed with brine (40 mL×2). The separated organic layer was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (EA / PE=1 / 5) to give 5-bromopyrazolo[1,5-a]pyridine-2,3-dicarboxylic acid dimethyl ester (200 mg, 13% yield) as a yellow solid. MS (ESI): 313.0, 315.0 [M+H] + .

[0686] Step 3) 5-bromo-3-(methoxycarbonyl)pyrazolo[1,5-a]pyridine-2-carboxylic acid

[0687] To a solution of 5-bromopyrazolo[1,5-a]pyridine-2,3-dicarboxylic acid dimethyl ester (200 mg, 0.19 mmol) in THF (10 mL) was added a solution of NaOH (26 mg, 0.64 mmol) in H2O (3 mL), and the mixture was stirred at room temperature for 16 h. The mixture was concentrated in vacuo, the residue was diluted with water (10 mL), and acidified with 1N HCl to a pH of 4-5. The precipitate was filtered, the filter cake was washed with water, and dried in vacuo to give 5-bromo-3-(methoxycarbonyl)pyrazolo[1,5-a]pyridine-2-carboxylic acid (180 mg, yield 89%) as a white solid. MS (ESI): 299.0, 301.0 [M+H] + .

[0688] Step 4) 5-bromo-2-((tert-butoxycarbonyl)amino)pyrazolo[1,5-a]pyridine-3-carboxylic acid methyl ester

[0689] Triethylamine (233 mg, 2.3 mmol) and DPPA (254 mg, 0.92 mmol) were added to a solution of 5-bromo-3-(methoxycarbonyl)pyrazolo[1,5-a]pyridine-2-carboxylic acid (230 mg, 0.77 mmol) in dioxane (20 mL), and the mixture was stirred at room temperature for 1 h. Then t-BuOH (114 mg, 1.53 mmol) was added, the temperature was raised to 100°C, and stirred for 16 h. After cooling to room temperature, the mixture was concentrated in vacuo. The residue was diluted with EtOAc (60 mL) and washed with brine (40 mL x 2). The separated organic layer was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (EA / PE=1 / 5) to give 5-bromo-2-((tert-butoxycarbonyl)amino)pyrazolo[1,5-a]pyridine-3-carboxylic acid methyl ester (150 mg, 53% yield) and 2-amino-5-bromopyrazolo[1,5-a]pyridine-3-carboxylic acid methyl ester (50 mg, 17.5% yield), both as white solids. MS (ESI): 392.0, 394.0 [M+H] + ,270.0,272.0[M+H] + Step 5) 2-amino-5-bromopyrazolo[1,5-a]pyridine-3-carboxylic acid

[0690] To a solution of methyl 2-amino-5-bromopyrazolo[1,5-a]pyridine-3-carboxylate (50 mg, 0.18 mmol) in MeOH (10 mL) was added an aqueous solution of NaOH (22 mg, 0.55 mmol) (2 mL), and the mixture was warmed to 50 ° C and stirred for 3 h. After cooling to room temperature, the mixture was concentrated in vacuo. The residue was diluted with water (5 mL) and acidified to pH = 4-5 with 1N HCl. It was then extracted with EtOAc (20 mL x 3), and the combined organic layers were washed with brine (20 mL x 2), dried over Na2SO4, and concentrated in vacuo to give 2-amino-5-bromopyrazolo[1,5-a]pyridine-3-carboxylic acid (45 mg, 90% yield) as a light solid. MS (ESI): 255.9, 257.9 [M+H] + .

[0691] Step 6) (S)-2-amino-5-bromo-N-(1-(8-((1-(methyl-d3)-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyridine-3-carboxamide

[0692] To a solution of (S)-3-(1-aminoethyl)-8-((1-(methyl-d3)-1H-pyrazol-4-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (65 mg, 0.18 mmol) and 2-amino-5-bromopyrazolo[1,5-a]pyridine-3-carboxylic acid (45 mg, 0.18 mmol) in DCM (30 mL) were added HOAt (32 mg, 0.23 mmol), EDCI (56 mg, 0.29 mmol) and DIEA (75 mg, 0.59 mmol), and the mixture was warmed to 40°C and stirred for 16 h. After cooling to room temperature, the mixture was diluted with DCM (30 mL) and washed with brine (30 mL x 2). The separated organic layer was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (DCM / MeOH=20 / 1) to give (S)-2-amino-5-bromo-N-(1-(8-((1-(methyl-d3)-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyridine-3-carboxamide (100 mg, 76% yield) as a yellow solid. MS (ESI): 609.0 [M+H] + .

[0693] Step 7) (S)-2-amino-5-cyano-N-(1-(8-((1-(methyl-d3)-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyridine-3-carboxamide

[0694] Under nitrogen atmosphere, Zn(CN)2 (19 mg, 0.16 mmol), Zn (11 mg, 0.16 mmol) and Pd(dppf)Cl2.DCM (7 mg, 0.008 mmol) were added to a solution of (S)-2-amino-5-bromo-N-(1-(8-((1-(methyl-d3)-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyridine-3-carboxamide (50 mg, 0.08 mmol) in DMF (5 mL), and the mixture was warmed to 100°C and stirred for 16 h. After cooling to room temperature, the mixture was diluted with EtOAc (60 mL) and washed with brine (30 mL×3). The separated organic layer was dried over Na2SO4 and concentrated in vacuo. The residue was purified by preparative HPLC (ACN-H2O, 0.1FA, gradient 40%-60%) to give (S)-2-amino-5-cyano-N-(1-(8-((1-(methyl-d3)-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyridine-3-carboxamide (7.8 mg, 17% yield) as a yellow solid. MS (ESI): 556.1 [M+H] + . 1 H NMR (400MHz, CD3OD) δ8.48(dd,J=6.8,0.8Hz,1H),8.18(d,J=0.8Hz,1H),7.87(s,1H),7.69(s,1H),7.67-7.52(m, 7H), 7.44 (d, J=7.6Hz, 1H), 7.04 (dd, J=7.2, 1.6Hz, 1H), 6.94 (s, 1H), 4.76 (q, J=6.8Hz, 1H), 1.50 (d, J=6.8Hz, 3H).

[0695] Example 39 (S)-2-amino-5-methoxy-N-(1-(8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyridine-3-carboxamide

[0696]

[0697] Step 1) 2-((tert-Butyloxycarbonyl)amino)-5-methoxypyrazolo[1,5-a]pyridine-3-carboxylic acid methyl ester

[0698] Pd2(dba)3 (25 mg, 0.027 mmol) and X-phos (26 mg, 0.054 mmol) were added to a solution of 5-bromo-2-((tert-butoxycarbonyl)amino)pyrazolo[1,5-a]pyridine-3-carboxylic acid methyl ester (100 mg, 0.27 mmol) and sodium methoxide (44 mg, 0.81 mmol) in dioxane (10 mL) and MeOH (5 mL), and the mixture was warmed to 70 ° C and stirred for 16 h. After cooling to room temperature, it was concentrated in vacuo. The residue was diluted with EtOAc (50 mL) and washed with brine (30 mL x 2). The separated organic layer was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (EA / PE=1 / 2) to give methyl 2-((tert-butoxycarbonyl)amino)-5-methoxypyrazolo[1,5-a]pyridine-3-carboxylate (80 mg, 73.7% yield) as a yellow solid. MS (ESI): 344.1 [M+Na] + .

[0699] Step 2) 2-amino-5-methoxypyrazolo[1,5-a]pyridine-3-carboxylic acid methyl ester

[0700] TFA (1 mL) was added to a solution of methyl 2-((tert-butoxycarbonyl)amino)-5-methoxypyrazolo[1,5-a]pyridine-3-carboxylate (80 mg, 0.25 mmol) in DCM (3 mL), and the mixture was stirred at room temperature for 2 h. The mixture was concentrated in vacuo to give methyl 2-amino-5-methoxypyrazolo[1,5-a]pyridine-3-carboxylate (60 mg, yield 87%) as a yellow oil. MS (ESI): 222.1 [M+H] + .

[0701] Step 3) 2-amino-5-methoxypyrazolo[1,5-a]pyridine-3-carboxylic acid

[0702] To a solution of 2-amino-5-methoxypyrazolo[1,5-a]pyridine-3-carboxylic acid methyl ester (60 mg, 0.27 mmol) in MeOH (10 mL) was added an aqueous solution of NaOH (32 mg, 0.81 mmol) (3 mL), and the mixture was heated to 65 ° C and stirred for 48 h. After cooling to room temperature, it was concentrated in vacuo. The residue was diluted with water (3 mL) and acidified to pH = 4-5 with 1N HCl, and the precipitate was collected by filtration. The filter cake was washed with water (2 mL) and dried in vacuo to obtain 2-amino-5-methoxypyrazolo[1,5-a]pyridine-3-carboxylic acid (50 mg, yield 53%) as a yellow solid. MS (ESI): 208.1 [M + H] + .

[0703] Step 4) (S)-2-amino-5-methoxy-N-(1-(8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyridine-3-carboxamide

[0704] To a solution of (S)-3-(1-aminoethyl)-8-((1-(methyl-d3)-1H-pyrazol-4-yl)ethynyl)-2-phenylisoquinolin-1(2H)-one (89 mg, 0.24 mmol) and 2-amino-5-methoxypyrazolo[1,5-a]pyridine-3-carboxylic acid (50 mg, 0.24 mmol) in DCM (40 mL) were added EDCI (69 mg, 0.36 mmol), HOAt (49 mg, 0.36 mmol) and DIEA (93 mg, 0.72 mmol), and the mixture was warmed to 40°C and stirred for 16 h. After cooling to room temperature, the mixture was diluted with DCM (50 mL) and washed with brine (30 mL x 2). The separated organic layer was dried over Na2SO4 and concentrated in vacuo. The residue was purified by preparative HPLC (ACN-H2O, 0.1FA, gradient 40%-60%) to give (S)-2-amino-5-methoxy-N-(1-(8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyridine-3-carboxamide (6.5 mg, 5% yield) as a white solid. MS (ESI): 561.2 [M+H] + . 1 H NMR (400MHz, CD3OD) δ8.15(d,J=7.6Hz,1H),7.86(s,1H),7.69(d,J=3.2Hz,1H),7.67–7.53(m,7H),7.43(d,J=7.6Hz,1H), 7.10(d,J=2.4Hz,1H),6.94(s,1H),6.55(dd,J=7.6,2.7Hz,1H),4.74(q,J=6.8Hz,1H),3.90(s,3H),1.47(d,J=6.8Hz,3H).

[0705] Biological tests

[0706] Kinase Activity Assay The activity of the compounds of the present invention as PI3K and mTOR kinase inhibitors can be evaluated by the following assay. General Description of Kinase Assay The kinase assay is performed by detecting myelin basic protein (MBP) incorporating γ-33P-ATP. Prepare 20 μg / mL of MBP (Sigma #M-1891) in tris-buffered saline (TBS; 50 mM Tris pH 8.0, 138 mM NaCl, 2.7 mM KCl) and coat a high-binding white 384-well plate (Greiner) with 60 μL per well. Incubate at 4°C for 24 hours. Wash the plate three times with 100 μL TBS. Kinase reactions were performed in a total volume of 34 μL of kinase buffer (5 mM Hepes pH 7.6, 15 mM NaCl, 0.01% bovine serum albumin (Sigma #I-5506), 10 mM MgCl2, 1 mM DTT, 0.02% TritonX-100). Compounds were dissolved in DMSO and added to each well, with a final DMSO concentration of 1%. Each data was measured twice, and at least two experiments were performed for each compound. For example, the final concentration of the enzyme was 10 nM or 20 nM. The reaction was started by adding unlabeled ATP (10 μM) and γ-33P labeled ATP (2×106 cpm per well, 3000 Ci / mmole). The reaction was shaken at room temperature for 1 hour. The 384-well plate was washed 7x with PBS, and then 50 μL of scintillation fluid was added to each well. A Wallac Trilux counter can also be used.

[0707] The above test method can obtain the IC of inhibition 50 and / or inhibition constant Ki. IC 50 Defined as the concentration of compound that inhibits 50% of enzyme activity under the test conditions. Use 1 / 2log dilution multiples to create a curve containing 10 concentration points and estimate IC 50 Values ​​(e.g., a typical curve is made through the following compound concentrations: 10 μM, 3 μM, 1 μM, 0.3 μM, 0.1 μM, 0.03 μM, 0.01 μM, 0.003 μM, 0.001 μM and 0 μM).

[0708] General PI3 Kinase Assay Protocol

[0709] PI3K (p110α / p85α) (h) [non-radioactive assay]

[0710] PI3K (p110α / p85α) (h) was incubated in a buffer solution containing 10 μM phosphatidylinositol-4,5-bisphosphate and MgATP (concentration determined according to requirements). The reaction was initiated after the addition of ATP solution. After incubation at room temperature for 30 minutes, the reaction was terminated by adding a stop solution containing EDTA and biotin phosphatidylinositol-3,4,5-triphosphate. Finally, the detection buffer was added, including Europium-labeled anti-GST monoclonal antibody, GST-labeled GRP1PH domain and streptavidin-allophycocyanin. The plate was read in time-resolved fluorescence mode, and the homogeneous time-resolved fluorescence (HTRF) signal was determined by the equation HTRF = 10000 × (Em665nm / Em620 nm).

[0711] PI3K (p110β / p85α) (h) [non-radioactive assay]

[0712] PI3K (p110β / p85α) (h) was incubated in a buffer solution containing 10 μM phosphatidylinositol-4,5-bisphosphate and MgATP (concentration determined according to requirements). The reaction was initiated after the addition of ATP solution. After incubation at room temperature for 30 minutes, the reaction was terminated by adding a stop solution containing EDTA and biotin phosphatidylinositol-3,4,5-triphosphate. Finally, the detection buffer was added, including Europium-labeled anti-GST monoclonal antibody, GST-labeled GRP1PH domain and streptavidin-allophycocyanin. The plate was read in time-resolved fluorescence mode, and the homogeneous time-resolved fluorescence (HTRF) signal was determined by the equation HTRF = 10000 × (Em665nm / Em620 nm).

[0713] PI3K (p110δ / p85α) (h) [Non-radioactive assay]

[0714] PI3K (p110δ / p85α) (h) was incubated in a buffer solution containing 10 μM phosphatidylinositol-4,5-bisphosphate and MgATP (concentration determined according to requirements). The reaction was initiated after the addition of ATP solution. After incubation at room temperature for 30 minutes, the reaction was terminated by adding a stop solution containing EDTA and biotin phosphatidylinositol-3,4,5-triphosphate. Finally, the detection buffer was added, including Europium-labeled anti-GST monoclonal antibody, GST-labeled GRP1PH domain and streptavidin-allophycocyanin. The plate was read in time-resolved fluorescence mode, and the homogeneous time-resolved fluorescence (HTRF) signal was determined by the equation HTRF = 10000 × (Em665nm / Em620 nm).

[0715] PI3K(p120γ)(h)[Non-radioactive assay]

[0716] PI3K (p120γ) (h) was incubated in a buffer solution containing 10 μM phosphatidylinositol-4,5-bisphosphate and MgATP (concentration determined according to requirements). The reaction was initiated after the addition of ATP solution. After incubation at room temperature for 30 minutes, the reaction was terminated by adding a stop solution containing EDTA and biotin phosphatidylinositol-3,4,5-triphosphate. Finally, the detection buffer was added, including Europium-labeled anti-GST monoclonal antibody, GST-labeled GRP1PH domain and streptavidin-allophycocyanin. The plate was read in time-resolved fluorescence mode, and the homogeneous time-resolved fluorescence (HTRF) signal was determined by the equation HTRF = 10000 × (Em665 nm / Em620 nm).

[0717] The kinase assay in the present invention was performed by Eurofins (Eurofins Cerep SA, Le Bois L'Evêque, 86600 Celle L'Evescault, France). The test results are shown in Table 1, where +: >100 nM; ++: 50-100 nM; +++: 10-50 nM; ++++: <10 nM.

[0718] Table 1. Kinase inhibition data of compounds of the present invention

[0719]

[0720]

[0721] The test results show that the compounds of the present invention have good inhibitory activity against PI3Kγ kinase and have high selectivity against other subtypes.

[0722] Cell viability assay

[0723] The cellular activity of the compounds of the present invention as PI3K kinase inhibitors can be evaluated by the following assay.

[0724] General description of cell-based assays:

[0725] PI3K-α, γ isoforms: First, the compound was diluted from the stock concentration to 5mM with 100% DMSO. The second step was to dilute the 5mM compound as the first point with 100% DMSO 4-fold for 10 points. The third step was to dilute 250-fold with serum-free medium, at which point the DMSO concentration was 0.4%. Then 50μL of the compound diluted with medium was transferred to a 50μL cell plate, at which point the DMSO concentration was 0.2%, and the final concentration of the compound was 10000nM, 2500nM, 625nM, 156.25nM, 39.06nM, 9.77nM, 2.44nM, 0.61nM, 0.15nM, 0.04nM.

[0726] PI3K-β, δ isoforms: First, dilute the compound from the stock concentration to 1.25mM with 100% DMSO. Second, dilute the 1.25mM compound as the first point with 100% DMSO 4 times for 10 points. Third, dilute 35.714 times with serum-free medium, then transfer 2.5μL of the compound diluted with medium to a 30μL cell plate, incubate in the incubator for 1 hour, and then add 2.5μL anti-IgM. At this time, the DMSO concentration is 0.2%, and the final concentration of the compound is 2500nM, 625nM, 156.25nM, 39.06nM, 9.77nM, 2.44nM, 0.61nM, 0.15nM, 0.04nM, 0.01nM.

[0727] PI3K-α inhibitory activity detection method

[0728] SKOV-3 cells were seeded at a density of 60,000 cells / 50 μL / well in a cell culture grade 96-well plate. The cell culture medium was serum-free RPMI-1640. The cells were cultured overnight at 37°C, 5% CO2 incubator. 50 μL / well of the test compound was added to the cells and cultured at 37°C, 5% CO2 incubator for 60 minutes. The final DMSO concentration was 0.2%. The culture medium was aspirated and 50 μL of 1x lysis buffer was added to each well. Shake at room temperature for 45 minutes. 16 μL of lysate was transferred to a 384 plate and 4 μL of the premixed antibody of Cisbio's Phospho-AKT (Ser473) kit was added. Centrifuge at 1000 rpm / min for one minute and incubate at 22°C for 4 hours before reading with Spark (665nm / 615nm).

[0729] PI3K-β inhibitory activity detection method

[0730] 786-O cells were seeded into a 96-well plate at a density of 30,000 cells / 50 μL / well. The cell culture medium was RPMI-1640 without serum. The cells were cultured overnight at 37°C, 5% CO2 incubator. 50 μL / well of the compound to be tested was added to the cells and cultured in a 37°C, 5% CO2 incubator for 60 minutes. The final DMSO concentration was 0.2%. The culture medium was aspirated and 50 μL of 1x lysis buffer was added to each well. The cells were shaken at room temperature for 45 minutes. 16 μL of the lysate was transferred to a 384 plate and 4 μL of the premixed antibody of the Phospho-AKT (Ser473) kit of Cisbio was added. The cells were centrifuged at 1000 rpm / min for one minute and incubated at 22°C for 4 hours before reading with Envision (665nm / 615nm).

[0731] PI3K-δ inhibitory activity detection method

[0732] Raji cells were cultured in 96-well plates at 30 μL per well, 50,000 cells, and the cell culture medium was serum-free RPMI-1640. The cells were incubated overnight in a 5% CO2 and 37°C incubator. After 18 hours of serum starvation, 2.5 μL of compound (14X) was added to the cells and incubated in the incubator for 60 minutes. Then 2.5 μL (14X, diluted with serum culture medium) of anti-human IgM (Jackson Immuno Research) was added and placed in the incubator for stimulation for 30 minutes (final concentration of 10 μg / mL). 11.5 μL of 4x lysis buffer was added to each well. Shake at room temperature for 45 minutes. 16 μL of lysate was added to a 384 plate, and 4 μL of the premixed antibody from the Phospho-AKT (Ser473) kit from Cisbio was added. Centrifuge at 1000 rpm / min for one minute, incubate at 22°C for 4 hours, and read using Spark (665nm / 615nm).

[0733] PI3K-γ inhibitory activity detection method

[0734] Resuspend RAW264.7 cells in serum-free DMEM medium and add 60,000 cells / 45 μL of cell suspension to each well of a 96-well plate. Incubate the cells in a 5% CO2, 37°C incubator overnight. After 18 hours of serum starvation, add 50 μL of compound and incubate in the incubator for 60 minutes. Then add 5 μL of 25nM C5a (R&D Systems, diluted with serum medium) to stimulate for 5 minutes. Aspirate the medium and add 50 μL of 1x lysis buffer to each well. Shake at room temperature for 45 minutes. Transfer 16 μL of lysate to a 384 plate and add 4 μL of premixed antibodies from Cisbio's Phospho-AKT (Ser473) kit. Centrifuge at 1000 rpm / min for one minute at 22°C. After incubation for 4 hours, read with Spark (665nm / 615nm).

[0735] The kinase assay in the present invention was performed by BioDuro Biotech (Jiangsu) Co., Ltd. The test results are shown in Table 2, where +: >500nM; ++: 100-500nM; +++: 50-100nM; ++++: <50nM.

[0736] The synthesis of the positive drug IPI-549 refers to patent WO2015143012A1. Its chemical structure is as follows:

[0737]

[0738] Table 2 Cellular activity data of the compounds of the present invention

[0739]

[0740] The test results show that the compound of the present invention has significant inhibitory activity against PI3Kγ and has very obvious selectivity against other PI3K subtypes. The compound of the present invention has higher selectivity than the positive drug IPI-549.

[0741] Study on the metabolic stability of the compounds of the present invention in human liver microsomes

[0742] The metabolic stability of the compounds of the present invention as PI3K kinase inhibitors can be evaluated by the following test.

[0743] General description of the liver microsome stability test:

[0744] The test compound was co-incubated with human liver microsomes, and NADPH was added to start the reaction. 30 μl was taken out at 0, 5, 15, 30 and 60 minutes and transferred to 300 μl of acetonitrile containing internal standard to terminate the reaction. Then vortex vigorously for 1 minute and centrifuge at 4000 rpm for 15 minutes below 4°C. After protein precipitation, 100 μl of the supernatant was taken out, diluted with 100 μl of distilled water and analyzed by LC-MS / MS method. The in vitro intrinsic clearance rate was calculated based on the elimination half-life of the test compound in the incubation system. Verapamil was used as a control compound and incubated in parallel for 5 minutes.

[0745] Data Analysis:

[0746] The remaining percentage (% Remaining) was calculated from the ratio of the peak area of ​​the test compound in the non-zero time point sample to the peak area of ​​the test compound in the zero time point sample. 1 / 2 , min) and in vitro intrinsic clearance (C Lint ,μL.min -1 .mg -1 ) is given by the following equation:

[0747]

[0748]

[0749]

[0750] The relevant parameters of the stability of the example compounds of the present invention and IPI-549 in human liver microsomes are shown in Table 3.

[0751] Table 3 Parameters related to the stability of the compounds of the present invention in human liver microsomes

[0752] Compound <![CDATA[Half-life (T 1 / 2 , min)]]> IPI-549 122.6 Embodiment 13 63.7 Example 16 154.7 Embodiment 21 77.2 Embodiment 28 656.9 Embodiment 37 94.7

[0753] The test results show that compared with IPI-549, the half-life of Examples 16 and 28 of the present invention is significantly prolonged, and the stability of human liver microsomes is significantly improved. Comparing Examples 16 and 28 of the present invention, when the hydrogen of the methyl group on the pyrazole is replaced by deuterium, the stability of human liver microsomes is significantly improved; similarly, comparing Examples 21 and 37 of the present invention, when the hydrogen of the methyl group on the pyrazole is replaced by deuterium, the stability of human liver microsomes is also significantly improved. Oral pharmacokinetics of the compound CD-1 of the present invention in mice

[0754] The pharmacokinetic study of the compounds of the present invention as PI3K kinase inhibitors can be evaluated by the following test.

[0755] General description of pharmacokinetic studies:

[0756] CD-1 mice were used as test animals, and the LC-MS / MS method was used to determine the drug concentration in plasma at different times after oral administration of the compound of the present invention. Intravenous injection dosage and formulation: 2 mg / kg, 5% NMP / 40% PEG400 / 55% PBS; oral dosage and formulation: 10.0 mg / kg, 0.5% (w / v) hydroxymethyl cellulose and 0.05% Tween 80 aqueous solution. Weigh an appropriate amount of the compound to be tested, add an appropriate amount of 0.5% (w / v) hydroxymethyl cellulose and 0.05% Tween 80 aqueous solution, vortex and sonicate, and finally prepare a test solution with a concentration of 1 mg / mL. The sampling time of the intravenous injection group is: 0.0833, 0.25, 0.5, 1, 2, 4, 8, 24h after administration. The sampling time of the oral group is: 0.25, 0.5, 1, 2, 4, 8, 24h. Finally, the LC-MS / MS method was used to determine the content of the test compound in the plasma of CD-1 mice after intravenous injection and oral administration of different compounds.

[0757] Table 4 Pharmacokinetic parameters of the compounds of the present invention

[0758]

[0759] The test results show that Example 16 and Example 37 of the present invention have good pharmacokinetic absorption in CD-1 mice, a long half-life, high bioavailability, and both have good pharmacokinetic properties.

[0760] Finally, it should be noted that there are other ways to implement the present invention. Accordingly, the embodiments of the present invention are described as examples, but are not limited to the contents described in the present invention, and may also be modified within the scope of the present invention or added to the claims. All publications or patents cited in the present invention will be used as references of the present invention.

Claims

1. A compound having a structure represented by formula (I): or a pharmaceutically acceptable salt; in, X is -C(R c )-, or N; Z 1 is N; Z 2 -C(R 4d )-or N; R a C 1-3 alkyl; R b and R c Each independently is H; R 1 yes Where R 1 Optionally 0, 1 or 2 R 5 replace; R 2 At each occurrence, each is independently H or F; R 3 is phenyl; wherein R 3 Optionally, 0, 1, 2, 3, 4 or 5 R 6 replace; R 4a H, D, F, Cl, -CN, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy, C 1-6 Hydroxyalkyl or C 3-8 Cycloalkyl; wherein R 4a is independently optionally substituted with 0, 1, 2, 3, 4 or 5 D; R 4b and R 4c Each independently represents H, D, F, Cl, C 1-3 Alkyl, C 1-3 Deuterated alkyl or C 1-3 Haloalkyl; R 4d is H; R 5 H, D, C 1-3 Alkyl or C 1-3 Deuterated alkyl; R 6 is H or D; n is 0 or 1; The conditions are: 1) When R 1 When it is pyrazolyl, R 1 Optionally 0, 1 or 2 R 5 Replace, then each R 2 , R 4a , R 4b and R 4c At least one of them is independently F, Cl, -CN, C 1-6 Alkoxy, C 1-6 Deuterated alkyl, C 1-6 Deuterated alkoxy, C substituted with 1 to 5 deuterium 1-6 Haloalkyl, C substituted with 1 to 5 deuterium 1-6 Hydroxyalkyl, or C substituted by 1 to 5 deuterium 3-8 Cycloalkyl.

2. The compound according to claim 1, which has the structure shown in formula (Ia): or a pharmaceutically acceptable salt.

3. The compound according to any one of claims 1 to 2, wherein R 4a H, D, F, Cl, C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Haloalkyl, C 1-3 Halogenated alkoxy, C 1-3 Hydroxyalkyl or C 3-6 Cycloalkyl; wherein R 4a is independently optionally substituted with 0, 1, 2, 3, 4 or 5 D.

4. The compound according to any one of claims 1 to 2, wherein R 4a H, F, Cl, D, -CN, -OCH3, -OCH2CH3, 5. A compound having the structure shown in formula (IIa): or a pharmaceutically acceptable salt; in, X is -C(R c )-, or N; R a C 1-6 alkyl; R b and R c Each independently is H; R 1 yes R 2 At each occurrence, is independently H, F, Cl, Br, I or -OH; R 3 is phenyl; wherein R 3 Optionally, 0, 1, 2, 3, 4 or 5 R 6 replace; R 4 At each occurrence, it is independently H or D; R 6 is H or D; m is 0, 1, 2, or 3; and n is 0, 1, 2, or 3.

6. The compound according to any one of claims 1 to 2, wherein R 5 At each occurrence, is independently H, D, methyl, ethyl or -CD3.

7. The compound according to any one of claims 1 to 2 and 5, wherein R a It is methyl, ethyl or isopropyl.

8. The compound according to claim 1, which is a compound having one of the following structures: or a pharmaceutically acceptable salt.

9. The compound according to claim 5, which is a compound having one of the following structures: or a pharmaceutically acceptable salt.

10. A pharmaceutical composition comprising the compound or pharmaceutically acceptable salt according to any one of claims 1 to 9, and a pharmaceutically acceptable excipient, diluent or carrier, or a combination thereof.

11. The pharmaceutical composition according to claim 10, further comprising an additional therapeutic agent.

12. Use of a compound according to any one of claims 1 to 9 or a pharmaceutical composition according to any one of claims 10 to 11 in the preparation of a medicament for preventing, treating, curing, and / or alleviating a disease, disorder, and / or condition mediated by PI3-kinase, or inhibiting PI3-kinase activity.

13. The use according to claim 12, wherein the PI3-kinase mediated disease, disorder, and / or condition is selected from respiratory disease, viral infection, non-viral respiratory infection, allergic disease, autoimmune disease, inflammatory disease, cardiovascular disease, hematological malignancy, neurodegenerative disease, pancreatitis, multiple organ failure, kidney disease, platelet aggregation, cancer, transplant rejection, graft rejection, lung injury or pain.

14. The use according to claim 12, wherein the PI3-kinase mediated disease, disorder, and / or condition is selected from asthma, chronic obstructive pulmonary disease (COPD), viral respiratory infection, viral respiratory disease exacerbation, aspergillosis, leishmaniasis, allergic rhinitis, allergic dermatitis, rheumatoid arthritis, multiple sclerosis, inflammatory bowel disease, thrombosis, atherosclerosis, hematological malignancies, neurodegenerative diseases, pancreatitis, multiple organ failure, kidney disease, platelet aggregation, cancer, transplant rejection, graft rejection, lung injury, pain associated with rheumatoid arthritis or osteoarthritis, back pain, systemic inflammatory pain, retrohepatic neuralgia, diabetic neuropathy, inflammatory neuropathic pain, trigeminal neuralgia or central pain.

15. The use according to claim 13 or 14, wherein the cancer is selected from acute myeloid leukemia, myelodysplastic syndrome, myeloproliferative disease, chronic myeloid leukemia, T-cell acute lymphoblastic leukemia, B-cell acute lymphoblastic leukemia, non-Hodgkin's lymphoma, B-cell lymphoma, solid tumors, or breast cancer.

16. The use according to claim 12, wherein the PI3-kinase is PI3K-γ.

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