CN116514817A - Heterocyclic compound with AKT kinase inhibitory activity, preparation method and medical use thereof - Google Patents

Abstract

The invention relates to a heterocyclic compound with AKT kinase inhibition activity, a preparation method and medical application thereof. In particular, the invention relates to a compound shown in a general formula (I), a preparation method thereof, a pharmaceutical composition containing the compound and application of the compound as an AKT kinase inhibitor in medicines for preventing and/or treating abnormal cell growth such as cancers. The definition of each group in the general formula (I) is the same as that in the specification.

Description

Heterocyclic compounds with AKT kinase inhibitory activity and preparation methods and medical uses thereof

Technical Field

The present invention belongs to the field of medical technology and relates to a pyrimidine heterocyclic compound, a preparation method thereof and a pharmaceutical composition containing the pyrimidine heterocyclic compound, and use of the pyrimidine heterocyclic compound as an AKT kinase inhibitor for treating abnormal cell growth such as cancer.

Background Art

AKT, also known as protein kinase B (PKB), is an important member of the PI3K/Akt/mTOR signaling pathway, which affects biological processes such as development, glucose homeostasis, tumor growth and metastasis by regulating numerous downstream effector molecules. Overactivation of the PI3K/AKT pathway is found in more than 50% of tumors, such as breast cancer, prostate cancer, and pancreatic cancer. Given the role of AKT as a key signaling center for tumor survival, people have been working hard to develop drugs targeting AKT kinase for many years.

AKT is divided into three subtypes, AKT1, AKT2, and AKT3. AKT1 and AKT2 are distributed throughout the body, while AKT3 is mainly expressed in some organs such as the brain, breast, heart, and kidney (Masure et al., Eur J Biochem, 1999: 265 (1), 353-360; Yang et al., J Biol Chem, 2003: 278 (34), 32124-32131). All three subtypes belong to the AGC kinase family, and the homology of the kinase domain is more than 85% (Hanks et al., Faseb J, 1995: 9 (8), 576-596). Structurally, they are composed of three parts: the amino-terminal PH domain, the ATP-binding kinase domain (CAT), and the carboxyl-terminal domain containing a hydrophobic motif (HM) (Kumar et al., Oncogene, 2005: 24 (50), 7493-7501). The activity of AKT is regulated by phosphorylation and dephosphorylation, and depends on the change of AKT conformation. In the absence of signal stimulation, the PH domain and CAT domain of AKT are connected by intramolecular interactions, thereby maintaining the inactive state of AKT in the cytoplasm (PH-In conformation) (Calleja et al., PLoS Biol, 2007: 5 (4), e95; Yudushkin et al., IUBMB Life, 2020: 72 (6), 1115-1125). On the cell membrane surface, RTKs or G protein-coupled receptors (GPCRs) activate PI3K, which then phosphorylates PIP2 and converts it into PIP3; PIP3 binds to the PH domain and activates AKT through PDK1-mediated phosphorylation, causing AKT to undergo conformational changes (PH-out conformation) and translocate to the plasma membrane. The PH-out conformation exposes the CAT and regulatory domains, leading to phosphorylation of two main sites: threonine residues in the CAT domain (Thr308 of AKT1, Thr309 of AKT2, and Thr305 of AKT3) and serine residues in the HM domain (Chu et al., Elife, 2020: 3(9), e59151; Lucic et al., Proc Natl Acad Sci USA, 2018: 115(17), E3940-E3949). In addition, ATP occupies the AKT nucleic acid binding pocket, slowing down the dephosphorylation of AKT and promoting full activation (Chan et al., Cell Cycle, 2012: 11(3), 475-478). Phosphorylated AKT activates the downstream effector molecule mTOR through TSC1/2, upregulating multiple transcription factors, promoting protein translation, cell growth, survival, etc. PIK3CA amplification/mutation (Shimoi et al., Cancer Sci, 2018: 109(8), 2558-2566), PTEN loss (Milella et al., Front Oncol, 2015: 5(24)), AKT mutations (such as E17K mutations in the PH domain) (Kalinsky et al., JAMA Oncol, 2021: 7(2), 271-278), and overactivation of other RTKs can lead to sustained activation of AKT signaling. AKT deactivation is mainly controlled by phosphatases such as PTEN, PP2A, PHLPP, and SHIP (Luongo et al., Cancers, 2019: 11(8); Villalobos-Ayala et al., Cancers, 2020: 12(12); Grzechnik et al., Biochem Soc Trans, 2016: 44(6), 1675-1682).

AKT kinase has been discovered for more than 30 years, and no drug has been successfully marketed. Although many PI3K inhibitors upstream of AKT have been approved for the treatment of tumors, both pan-PI3K inhibitors and selective PI3K subtype inhibitors have a high toxicity problem; and the negative feedback mechanism that occurs after inhibiting downstream mTOR will cause pAKT levels to re-upregulate, thereby activating other downstream carcinogenic effector factors. Therefore, the development of ideal molecules that directly target AKT has the hope of avoiding the serious side effects caused by inhibiting upstream PI3K, and can also avoid the negative feedback mechanism caused by inhibiting downstream mTOR.

According to the difference between the inhibitor and AKT binding site, drugs can be divided into three categories: PH domain inhibitors, allosteric inhibitors and ATP competitive inhibitors. The allosteric inhibitor MK-2206 is the first AKT inhibitor to enter clinical trials. Early clinical results showed good AKT signal inhibition effect and good tolerability (Yap et al., J Clin Oncol, 2011: 29 (35), 4688-4695), but due to serious toxic side effects, it has been removed from the pipeline of the research and development company. Currently, the main research is still based on ATP competitive inhibitors, such as capIIIAasertib (AZD5363) and ipatasertib (GDC0068) Phase III clinical trials have been conducted in HR+ and triple-negative breast cancer. A recent phase III result of ipatasertib showed that the combination of ipatasertib with abiraterone and prednisolone significantly improved radiographic progression-free survival in patients with PTEN-deficient, metastatic, and castration-resistant prostate cancer compared with placebo plus abiraterone (Sweeney et al., Lancet, 2021: 398(10295), 131-142). In addition, other AKT inhibitors such as afuresertib, uprosertib, and HZB0071 are also in clinical development.

Clinical research on AKT inhibitors still faces many challenges: low bioavailability, narrow therapeutic window, and side effects such as diarrhea and rash, as well as hyperglycemia, which is due to the fact that targeted inhibition of the PI3K/AKT pathway disrupts glucose uptake and metabolism in certain tissues, leading to hyperglycemia (Fruman et al., Cell, 2017: 170(4), 605-635). In addition, inhibition of liver AKT may cause liver damage, inflammation and carcinogenesis in mice, and promote lung metastasis (Wang et al., Cancer Cell, 2016: 29(4), 523-535). Future research directions need to improve kinase selectivity and tolerance, while exploring reasonable drug combinations to overcome the problem of PI3K/AKT inhibitor resistance.

Summary of the invention

After intensive research, the inventors have designed and synthesized a series of pyrimidine compounds, which show AKT1/2/3 kinase inhibitory activity and can be developed as drugs for treating and/or preventing diseases related to AKT activity, such as cancer.

Therefore, the object of the present invention is to provide a compound represented by general formula (I) or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof,

in:

Ring A is selected from C ₃ -C ₁₂ cycloalkyl, 3 to 14 membered heterocyclyl, C ₆ -C ₁₄ aryl and 6 to 14 membered heteroaryl; wherein said C ₃ -C ₁₂ cycloalkyl, 3 to 14 membered heterocyclyl, C ₆ -C ₁₄ aryl and 6 to 14 membered heteroaryl are each independently optionally substituted by one or more R ¹ ;

L is a bond or -CH ₂ -;

Ring B is selected from a 5- to 8-membered saturated heterocyclic group containing 1-2 nitrogen atoms; the heterocyclic group is optionally substituted by one or more R ³ ;

G is selected from hydrogen, phenyl, 5-6 membered heteroaryl, wherein the phenyl, 5-6 membered heteroaryl is optionally substituted by one or more R ⁴ ;

Z is CH or N;

Each R ¹ is independently selected from hydrogen, halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, and the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl are optionally substituted with one or more groups selected from halogen, amino, nitro, cyano, hydroxyl, thiol, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl; or,

Any two R ¹ together with the atoms to which they are attached form a cycloalkyl, heterocyclyl, aryl and heteroaryl group, wherein the cycloalkyl, heterocyclyl, aryl and heteroaryl group are optionally substituted with one or more groups selected from halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, carboxyl, ester, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl;

Each R ² is independently selected from hydrogen, halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, and the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl are optionally substituted with one or more groups selected from halogen, amino, nitro, cyano, hydroxyl, thiol, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl; or,

Any two ^R2 together with the atoms to which they are attached form a cycloalkyl, heterocyclyl, aryl and heteroaryl group, wherein the cycloalkyl, heterocyclyl, aryl and heteroaryl groups are optionally substituted with one or more groups selected from halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, carboxyl, ester, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl groups;

Each ^R3 is independently selected from hydrogen, halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, and the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl are optionally substituted with one or more groups selected from halogen, amino, nitro, cyano, hydroxyl, thiol, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl; or,

Any two ^R3 together with the atoms to which they are attached form a cycloalkyl, heterocyclyl, aryl and heteroaryl group, wherein the cycloalkyl, heterocyclyl, aryl and heteroaryl group are optionally substituted with one or more groups selected from halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, carboxyl, ester, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl;

each R ⁴ is independently selected from hydrogen, halogen, nitro, cyano, hydroxyl, thiol, alkyl, alkenyl, alkynyl, -S(O) _p ^Ra , -N ^Ra R ^b , -OR ^a , -C(O) Ra, -C(O ⁾ N ^Ra R ^b , -S(O) _p N ^Ra R ^b , cycloalkyl, heterocyclyl, aryl, heteroaryl, and the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl are optionally substituted with one or more groups selected from halogen, amino, nitro, cyano, hydroxyl, thiol, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl;

^Ra and ^Rb are each independently selected from hydrogen, halogen, hydroxyl, thiol, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, and the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl are optionally substituted with one or more groups selected from halogen, amino, nitro, cyano, hydroxyl, thiol, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl; or,

^Ra and ^Rb together with the nitrogen atom to which they are attached form a nitrogen-containing heterocyclic group, which is optionally substituted with one or more groups selected from halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, carboxyl, ester, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl and heteroaryl;

n is 0, 1, 2 or 3;

p is 0, 1 or 2.

In a preferred embodiment, the compound represented by the general formula (I) according to the present invention or its stereoisomer, tautomer, mesomorph, racemate, enantiomer, diastereoisomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein ring A is selected from C ₃ -C ₁₂ cycloalkyl and 3 to 14-membered heterocyclic group, preferably 3 to 14-membered heterocyclic group, more preferably 3 to 6-membered monocyclic heterocyclic group, 6 to 11-membered bicyclic heterocyclic group, further preferably 6 to 10-membered fused heterocyclic group, 6 to 10-membered spiro heterocyclic group; the cycloalkyl or heterocyclic group is each independently optionally substituted by one or more R ¹ ;

Each R ¹ is independently selected from hydrogen, halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C 2 -C 6 alkenyl, C _{2 -C 6 alkynyl, C 3 -C 6 cycloalkyl} , 3 to 6 membered heterocyclyl, C ₆ -C _{10 aryl} , 5 to 10 membered heteroaryl, the C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclyl, C ₆ -C ₁₀ aryl, 5 to 10 membered heteroaryl optionally selected from halogen, amino, nitro, cyano, hydroxyl, thiol, carboxyl, ester, oxo, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C _{1 -C6} haloalkoxy, _C2 - _C6 alkenyl, _C2 - _C6 alkynyl, _C3 - _C6 cycloalkyl, 3- to 6-membered heterocyclyl, _C6 - _C10 aryl, or 5- to 10-membered heteroaryl; or,

Any two ^R1 together with the atoms to which they are attached form a cycloalkyl, heterocyclyl, aryl and heteroaryl group, and the cycloalkyl, heterocyclyl, aryl and heteroaryl groups are optionally substituted by one or more groups selected from halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, carboxyl, ester, _C1 - _C6 alkyl, _C1 - _C6 alkoxy, _{C1 - C6} haloalkyl, _C1 - _C6 haloalkoxy, _{C2-C6 alkenyl} , _C2 - _C6 alkynyl, C3- _C6 cycloalkyl, 3 to 6 membered heterocyclyl, _C6 - _C10 aryl and 5 to 10 membered heteroaryl.

In a specific embodiment, according to the compound represented by general formula (I) of the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereoisomer, or mixture thereof, or pharmaceutically acceptable salt thereof, ring A is a 3- to 6-membered monocyclic heterocyclic group; the 3- to 6-membered monocyclic heterocyclic group is optionally substituted by one or more ^R1 ; each ^R1 is independently selected from hydrogen, halogen, amino, cyano, oxo, _hydroxyl , thiol, _C1 - _C6 alkyl, _C1 - _C6 alkoxy, C1- _C6 haloalkyl, _C1 - _C6 haloalkoxy.

In another specific embodiment, according to the compound represented by the general formula (I) of the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereoisomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein ring A is a 6- to 11-membered fused heterocyclic group or a 6- to 11-membered spiro heterocyclic group, and the 6- to 11-membered fused heterocyclic group or the 6- to 11-membered spiro heterocyclic group is optionally substituted by one or more R ¹ ; each R ¹ is independently selected from hydrogen, halogen, amino, cyano, oxo, hydroxyl, thiol, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkoxy.

In another specific embodiment, according to the compound represented by the general formula (I) of the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereoisomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein ring A is a 6- to 11-membered spiroheterocyclic group, preferably a 7- to 10-membered spiroheterocyclic group, and the spiroheterocyclic group is optionally substituted by one or more ^R1 ; each ^R1 is independently selected from hydrogen, halogen, amino, cyano, oxo, hydroxyl, thiol, _C1 - _C6 alkyl, _C1 - _{C6 alkoxy, C1-C6 haloalkyl , C1} - _C6 haloalkoxy.

In another specific embodiment, according to the compound represented by the general formula (I) of the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereoisomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein ring A is a 6- to 11-membered fused heterocyclic group, preferably a 6- to 10-membered fused heterocyclic group, and the fused heterocyclic group is optionally substituted by one or more ^R1 ; each ^R1 is independently selected from hydrogen, halogen, amino, cyano, oxo, hydroxyl, thiol, _C1 - _C6 alkyl, _C1 - _C6 alkoxy, _C1 - _C6 haloalkyl, _C1 - _C6 haloalkoxy.

In a preferred embodiment, the compound represented by general formula (I) according to the present invention or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, is a compound represented by general formula (II) or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof,

in:

m is 0, 1, 2, 3 or 4;

Each R ¹ is independently selected from hydrogen, halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C 2 -C 6 alkenyl, C _{2 -C 6 alkynyl, C 3 -C 6 cycloalkyl} , 3 to 6 membered heterocyclyl, C ₆ -C ₁₀ aryl, 5 to ₁₀ membered heteroaryl, the C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclyl, C ₆ -C ₁₀ aryl, 5 to 10 membered heteroaryl optionally selected from halogen, amino, nitro, cyano, hydroxyl, thiol, carboxyl, ester, oxo, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkoxy, C ₃ -C ₆ cycloalkyl, 3- to 6-membered heterocyclyl, C ₆ -C ₁₀ aryl, or 5- to 10-membered heteroaryl; or,

Any two ^R1 together with the atoms to which they are attached form a cycloalkyl, heterocyclyl, aryl and heteroaryl group, wherein the cycloalkyl, heterocyclyl, aryl and heteroaryl group are optionally further substituted with one or more groups selected from halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, carboxyl, ester, _C1 - _C6 alkyl, _C1 - _C6 alkoxy, C1- _C6 haloalkyl, _{C1 - C6} haloalkoxy, _C2 - _C6 alkenyl, _C2 - _C6 alkynyl, _C3 - _C6 cycloalkyl, 3-6 membered heterocyclyl, _C6 - _C10 aryl and 5-10 membered heteroaryl;

Ring B, L, G, Z, R ² and n are as defined in the general formula (I).

In a preferred embodiment, according to the compound represented by general formula (I) or general formula (II) according to the present invention or its stereoisomer, tautomer, mesomorph, racemate, enantiomer, diastereoisomer, or mixture thereof, or pharmaceutically acceptable salt thereof, any two ^R1 together with the atom to which they are connected form a _C3 - _C6 cycloalkyl or a 3- to 6-membered heterocyclic group, and the _C3 - _C6 cycloalkyl or 3- to ₆ -membered heterocyclic group is optionally substituted by one or more groups selected from halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, carboxyl, ester, _C1 - _C6 alkyl, _{C1 - C6} alkoxy, _{C1 - C6} haloalkyl, _C1 - _{C6 haloalkoxy, C2-C6 alkenyl} , C2-C6 alkynyl, _C3 - _C6 cycloalkyl, 3- to 6-membered heterocyclic group, _C6 - _C10 aryl and 5- to 10-membered heteroaryl.

In another preferred embodiment, according to the compound represented by general formula (I) or general formula (II) or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein R ¹ is selected from hydrogen.

In a specific embodiment, the compound represented by the general formula (I) according to the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein ring A is selected from:

Ring A is optionally substituted by one or more R ¹ ; each R ¹ is independently selected from hydrogen, halogen, amino, cyano, oxo, hydroxyl, mercapto, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkoxy.

In a preferred embodiment, according to the compound represented by general formula (I) or general formula (II) of the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, wherein L is a bond.

In another preferred embodiment, the compound of formula (I) or (II) according to the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein L is -CH ₂ -.

In another preferred embodiment, the compound of general formula (I) according to the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, is a compound of general formula (III-1), general formula (III-2) or general formula (III-3) or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof,

in,

_X1 and _X2 are each independently selected from _-CH2- , -O-, -NH-;

n ₁ is 0 or 1;

n ₂ is 0, 1, or 2;

each R ^c is independently selected from halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, carboxyl, ester, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C _{1 -C 6 haloalkyl, C 1 -C 6 haloalkoxy ,} C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclyl, C ₆ -C ₁₀ aryl and 5 to 10 membered heteroaryl;

q is 0, 1 or 2, preferably 0 or 1, more preferably 0;

Ring B, G, Z, ^R2 and n are as defined in the general formula (I).

In another preferred embodiment, the compound of general formula (I) according to the present invention or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, is a compound of general formula (IV-1) or (IV-2) or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof,

in,

Ring _E1 is C ₃ -C ₆ cycloalkyl or 3 to 6-membered heterocyclic group;

Ring _E2 is C ₃ -C ₆ cycloalkyl or 3 to 6-membered heterocyclic group;

Each R ^e is independently selected from halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, carboxyl, ester, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C _{1 -C 6 haloalkyl, C 1 -C 6 haloalkoxy ,} C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclyl, C ₆ -C ₁₀ aryl and 5 to 10 membered heteroaryl;

t is 0, 1 or 2, preferably 0 or 1, more preferably 0;

Ring B, G, Z, ^R2 and n are as defined in the general formula (I).

In another preferred embodiment, according to the compound represented by general formula (I), general formula (II), general formula (III-1), general formula (III-2), general formula (III-3), general formula (IV-1) or general formula (IV-2) of the present invention, or its stereoisomer, tautomer, mesomorph, racemate, enantiomer, diastereoisomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein ring B is a 6- to 8-membered saturated heterocyclic group containing 1-2 nitrogen atoms; ring B is preferably selected from the following groups:

Best More preferred

The single wavy line indicates the position connected to the carbonyl group, and the double wavy line indicates the position connected to the pyrimidine.

Ring B is optionally substituted with one or more R ³ ;

Each ^R3 is independently selected from hydrogen, halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, _C1 - _C6 alkyl, _C1 - _C6 alkoxy, _C2 - _C6 alkenyl, C2- _C6 alkynyl, _C3 - _C6 cycloalkyl, ₃ to 6 membered heterocyclyl, _C6 - _C10 aryl, 5 to 10 membered heteroaryl, the _C1 - _C6 alkyl, _C1 - _C6 alkoxy, _C2 - _C6 alkenyl, _C2 - _C6 alkynyl, _C3 - _C6 cycloalkyl, 3 to 6 membered heterocyclyl, _C6 - _C10 aryl, 5 to 10 membered heteroaryl being optionally selected from halogen, amino, nitro, cyano, hydroxyl, thiol, carboxyl, ester, oxo, _C1 - _C6 alkyl, _C1 - _C6 alkoxy, _C1 - _C6 haloalkyl, _C1 -C ₆ haloalkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3- to 6-membered heterocyclyl, C ₆ -C ₁₀ aryl, or 5- to 10-membered heteroaryl;

Preferably R ³ is hydrogen.

In another preferred embodiment, the compound of general formula (I) according to the present invention or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, is a compound of general formula (IIIA), general formula (IIIB) or general formula (IIIC) or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof,

in,

_X1 and _X2 are each independently selected from _-CH2- , -O-, -NH-;

n ₁ is 0 or 1;

n ₂ is 0, 1, or 2;

each R ^c is independently selected from halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, carboxyl, ester, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C _{1 -C 6 haloalkyl, C 1 -C 6 haloalkoxy ,} C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclyl, C ₆ -C ₁₀ aryl and 5 to 10 membered heteroaryl;

q is 0, 1 or 2, preferably 0 or 1, more preferably 0;

Each ^R3 is independently selected from hydrogen, halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, _C1 - _C6 alkyl, _C1 - _C6 alkoxy, _C2 - _C6 alkenyl, C2- _C6 alkynyl, _C3 - _C6 cycloalkyl, ₃ to 6 membered heterocyclyl, _C6 - _C10 aryl, 5 to 10 membered heteroaryl, the _C1 - _C6 alkyl, _C1 - _C6 alkoxy, _C2 - _C6 alkenyl, _C2 - _C6 alkynyl, _C3 - _C6 cycloalkyl, 3 to 6 membered heterocyclyl, _C6 - _C10 aryl, 5 to 10 membered heteroaryl being optionally selected from halogen, amino, nitro, cyano, hydroxyl, thiol, carboxyl, ester, oxo, _C1 - _C6 alkyl, _C1 - _C6 alkoxy, _C1 - _C6 haloalkyl, _C1 -C ₆ haloalkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclyl, C ₆ -C ₁₀ aryl, 5 to 10 membered heteroaryl, or one or more groups; preferably R ³ is hydrogen;

r is 0, 1, 2, 3 or 4, preferably 0, 1 or 2, more preferably 0 or 1;

G, Z, ^R2 and n are as defined in the general formula (I).

In another preferred embodiment, the compound of general formula (I) according to the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, is a compound of general formula (IVA) or (IVB) or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof,

in,

Ring _E1 is C ₃ -C ₆ cycloalkyl or 3 to 6-membered heterocyclic group;

Ring _E2 is C ₃ -C ₆ cycloalkyl or 3 to 6-membered heterocyclic group;

Each R ^e is independently selected from halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, carboxyl, ester, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C _{1 -C 6 haloalkyl, C 1 -C 6 haloalkoxy ,} C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclyl, C ₆ -C ₁₀ aryl and 5 to 10 membered heteroaryl;

t is 0, 1 or 2, preferably 0 or 1, more preferably 0;

Each ^R3 is independently selected from hydrogen, halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, _C1 - _C6 alkyl, _C1 - _C6 alkoxy, _C2 - _C6 alkenyl, C2- _C6 alkynyl, _C3 - _C6 cycloalkyl, ₃ to 6 membered heterocyclyl, _C6 - _C10 aryl, 5 to 10 membered heteroaryl, the _C1 - _C6 alkyl, _C1 - _C6 alkoxy, _C2 - _C6 alkenyl, _C2 - _C6 alkynyl, _C3 - _C6 cycloalkyl, 3 to 6 membered heterocyclyl, _C6 - _C10 aryl, 5 to 10 membered heteroaryl being optionally selected from halogen, amino, nitro, cyano, hydroxyl, thiol, carboxyl, ester, oxo, _C1 - _C6 alkyl, _C1 - _C6 alkoxy, _C1 - _C6 haloalkyl, _C1 -C ₆ haloalkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclyl, C ₆ -C ₁₀ aryl, 5 to 10 membered heteroaryl, or one or more groups; preferably R ³ is hydrogen;

r is 0, 1, 2, 3 or 4, preferably 0, 1 or 2, more preferably 0 or 1;

G, Z, ^R2 and n are as defined in the general formula (I).

In another preferred embodiment, according to the compound represented by general formula (I), general formula (II), general formula (III-1), general formula (III-2), general formula (III-3), general formula (IV-1), general formula (IV-2), general formula (IIIA), general formula (IIIB), general formula (IIIC), general formula (IVA) or general formula (IVB) of the present invention, or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereoisomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein G is selected from phenyl, and the phenyl is optionally substituted by one or more R ⁴ ;

Each R ⁴ is independently selected from halogen, cyano, hydroxyl, mercapto, C ₁ -C ₆ alkyl, -NR ^a R ^b , -OR ^a , wherein the C ₁ -C ₆ alkyl is optionally substituted with halogen;

R ^a and R ^b are each independently selected from hydrogen and C ₁ -C ₆ alkyl; or,

^Ra and ^Rb together with the nitrogen atom to which they are attached form a 5-6 membered nitrogen-containing heterocyclic group, which is optionally substituted by one or more groups selected from halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, carboxyl, ester, _{C1 - C6} alkyl, C1- _C6 alkoxy, _C1 - _C6 haloalkyl, _C1 - _C6 haloalkoxy.

In another preferred embodiment, according to the compound represented by general formula (I), general formula (II), general formula (III-1), general formula (III-2), general formula (III-3), general formula (IV-1), general formula (IV-2), general formula (IIIA), general formula (IIIB), general formula (IIIC), general formula (IVA) or general formula (IVB) of the present invention, or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereoisomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein G is selected from 5- or 6-membered heteroaryl, preferably 6-membered heteroaryl, more preferably pyridyl, and the heteroaryl is optionally substituted by one or more R ⁴ ;

Each R ⁴ is independently selected from halogen, cyano, hydroxyl, mercapto, C ₁ -C ₆ alkyl, -NR ^a R ^b , -OR ^a , wherein the C ₁ -C ₆ alkyl is optionally substituted with halogen;

R ^a and R ^b are each independently selected from hydrogen and C ₁ -C ₆ alkyl; or,

^Ra and ^Rb together with the nitrogen atom to which they are attached form a 5-6 membered nitrogen-containing heterocyclic group, which is optionally substituted by one or more groups selected from halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, carboxyl, ester, _{C1 - C6} alkyl, C1- _C6 alkoxy, _C1 - _C6 haloalkyl, _C1 - _C6 haloalkoxy.

In another preferred embodiment, the compound of general formula (I) according to the present invention or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, is a compound of general formula (IIID), general formula (IIIE), general formula (IIIF) or general formula (IIID-1), general formula (IIIE-1), general formula (IIIF-1) or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof,

in,

_X1 and _X2 are each independently selected from _-CH2- , -O-, -NH-;

n ₁ is 0 or 1;

n ₂ is 0, 1, or 2;

each R ^c is independently selected from halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, carboxyl, ester, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C _{1 -C 6 haloalkyl, C 1 -C 6 haloalkoxy ,} C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclyl, C ₆ -C ₁₀ aryl and 5 to 10 membered heteroaryl;

q is 0, 1 or 2, preferably 0 or 1, more preferably 0;

Each ^R3 is independently selected from hydrogen, halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, _C1 - _C6 alkyl, _C1 - _C6 alkoxy, _C2 - _C6 alkenyl, C2- _C6 alkynyl, _C3 - _C6 cycloalkyl, ₃ to 6 membered heterocyclyl, _C6 - _C10 aryl, 5 to 10 membered heteroaryl, the _C1 - _C6 alkyl, _C1 - _C6 alkoxy, _C2 - _C6 alkenyl, _C2 - _C6 alkynyl, _C3 - _C6 cycloalkyl, 3 to 6 membered heterocyclyl, _C6 - _C10 aryl, 5 to 10 membered heteroaryl being optionally selected from halogen, amino, nitro, cyano, hydroxyl, thiol, carboxyl, ester, oxo, _C1 - _C6 alkyl, _C1 - _C6 alkoxy, _C1 - _C6 haloalkyl, _C1 -C ₆ haloalkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclyl, C ₆ -C ₁₀ aryl, 5 to 10 membered heteroaryl, or one or more groups; preferably R ³ is hydrogen;

r is 0, 1, 2, 3 or 4, preferably 0, 1 or 2, more preferably 0 or 1;

Each R ⁴ is independently selected from halogen, cyano, hydroxyl, mercapto, C ₁ -C ₆ alkyl, -NR ^a R ^b , -OR ^a , wherein the C ₁ -C ₆ alkyl is optionally substituted with halogen; preferably R ⁴ is selected from halogen;

R ^a and R ^b are each independently selected from hydrogen and C ₁ -C ₆ alkyl; or,

^Ra and ^Rb together with the nitrogen atom to which they are attached form a 5-6 membered nitrogen-containing heterocyclic group, wherein the 5-6 membered nitrogen-containing heterocyclic group is optionally substituted with one or more groups selected from halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, carboxyl, ester, _{C1 - C6} alkyl, _C1 - _C6 alkoxy, C1- _C6 haloalkyl, _C1 - _C6 haloalkoxy;

s is 0, 1, or 2;

^R2 and n are as defined in the general formula (I).

In another preferred embodiment, the compound of the general formula (I) according to the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, is a compound of the general formula (IVC), general formula (IVD), general formula (IVC-1) or general formula (IVD-1) or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof,

in,

Ring _E1 is C ₃ -C ₆ cycloalkyl or 3 to 6-membered heterocyclic group;

Ring _E2 is C ₃ -C ₆ cycloalkyl or 3 to 6-membered heterocyclic group;

Each ^Re is independently selected from halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, carboxyl, ester, _C1 - _C6 alkyl, _C1 - _C6 alkoxy, C1-C6 haloalkyl, _C1 - _C6 haloalkoxy, _{C2 - C6} alkenyl, _C2 - _C6 alkynyl, _C3 - _C6 cycloalkyl, 3- to ₆ -membered heterocyclyl, _C6 - _C10 aryl, and 5- to 10-membered heteroaryl;

t is 0, 1 or 2, preferably 0 or 1, more preferably 0;

Each ^R3 is independently selected from hydrogen, halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, _C1 - _C6 alkyl, _C1 - _C6 alkoxy, _C2 - _C6 alkenyl, C2- _C6 alkynyl, _C3 - _C6 cycloalkyl, ₃ to 6 membered heterocyclyl, _C6 - _C10 aryl, 5 to 10 membered heteroaryl, the _C1 - _C6 alkyl, _C1 - _C6 alkoxy, _C2 - _C6 alkenyl, _C2 - _C6 alkynyl, _C3 - _C6 cycloalkyl, 3 to 6 membered heterocyclyl, _C6 - _C10 aryl, 5 to 10 membered heteroaryl being optionally selected from halogen, amino, nitro, cyano, hydroxyl, thiol, carboxyl, ester, oxo, _C1 - _C6 alkyl, _C1 - _C6 alkoxy, _C1 - _C6 haloalkyl, _C1 -C ₆ haloalkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclyl, C ₆ -C ₁₀ aryl, 5 to 10 membered heteroaryl, or one or more groups; preferably R ³ is hydrogen;

r is 0, 1, 2, 3 or 4, preferably 0, 1 or 2, more preferably 0 or 1;

Each R ⁴ is independently selected from halogen, cyano, hydroxyl, mercapto, C ₁ -C ₆ alkyl, -NR ^a R ^b , -OR ^a , wherein the C ₁ -C ₆ alkyl is optionally substituted with halogen; preferably R ⁴ is selected from halogen;

R ^a and R ^b are each independently selected from hydrogen and C ₁ -C ₆ alkyl; or,

^Ra and ^Rb together with the nitrogen atom to which they are attached form a 5-6 membered nitrogen-containing heterocyclic group, wherein the 5-6 membered nitrogen-containing heterocyclic group is optionally substituted with one or more groups selected from halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, carboxyl, ester, _{C1 - C6} alkyl, _C1 - _C6 alkoxy, C1- _C6 haloalkyl, _C1 - _C6 haloalkoxy;

s is 0, 1, or 2;

^R2 and n are as defined in the general formula (I).

In a preferred embodiment, according to the compounds represented by general formula (IV-1), general formula (IVA), general formula (IVC), general formula (IVC-1) or their stereoisomers, tautomers, mesomers, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, wherein ring _E1 is _C3 - _C6 cycloalkyl, preferably cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.

In another preferred embodiment, according to the compounds represented by general formula (IV-2), general formula (IVB), general formula (IVD), general formula (IVD-1) or their stereoisomers, tautomers, mesomers, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, wherein ring _E2 is _C3 - _C6 cycloalkyl, preferably cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.

In another preferred embodiment, according to the compounds represented by general formula (IV-1), general formula (IV-2), general formula (IVA), general formula (IVB), general formula (IVC), general formula (IVD), general formula (IVC-1), general formula (IVD-1) or their stereoisomers, tautomers, racemates, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, wherein ^Re is hydrogen and t is 1.

In another preferred embodiment, according to the compounds represented by general formula (IV-1), general formula (IV-2), general formula (IVA), general formula (IVB), general formula (IVC), general formula (IVD), general formula (IVC-1), general formula (IVD-1) or their stereoisomers, tautomers, racemates, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, wherein t is 0.

In another preferred embodiment, the compound of general formula (I) according to the present invention or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, is a compound of general formula (V) or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof,

in:

Ring A is selected from C ₃ -C ₁₂ cycloalkyl and 3 to 14-membered heterocyclyl, preferably 3 to 14-membered heterocyclyl, more preferably 3 to 6-membered monocyclic heterocyclyl, 6 to 11-membered bicyclic heterocyclyl, further preferably 6 to 10-membered fused heterocyclyl, 6 to 10-membered spiro heterocyclyl;

Each R ^d is independently selected from hydrogen, halogen, amino, cyano, oxo, hydroxyl, mercapto, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkoxy;

t is 0, 1, or 2;

Each ^R3 is independently selected from hydrogen, halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, _C1 - _C6 alkyl, _C1 - _C6 alkoxy, _C2 - _C6 alkenyl, C2- _C6 alkynyl, _C3 - _C6 cycloalkyl, ₃ to 6 membered heterocyclyl, _C6 - _C10 aryl, 5 to 10 membered heteroaryl, the _C1 - _C6 alkyl, _C1 - _C6 alkoxy, _C2 - _C6 alkenyl, _C2 - _C6 alkynyl, _C3 - _C6 cycloalkyl, 3 to 6 membered heterocyclyl, _C6 - _C10 aryl, 5 to 10 membered heteroaryl being optionally selected from halogen, amino, nitro, cyano, hydroxyl, thiol, carboxyl, ester, oxo, _C1 - _C6 alkyl, _C1 - _C6 alkoxy, _C1 - _C6 haloalkyl, _C1 -C ₆ haloalkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclyl, C ₆ -C ₁₀ aryl, 5 to 10 membered heteroaryl, or one or more groups; preferably R ³ is hydrogen;

r is 0, 1, 2, 3 or 4, preferably 0, 1 or 2, more preferably 0 or 1;

Each R ⁴ is independently selected from halogen, cyano, hydroxyl, mercapto, C ₁ -C ₆ alkyl, -NR ^a R ^b , -OR ^a , wherein the C ₁ -C ₆ alkyl is optionally substituted with halogen; preferably R ⁴ is selected from halogen;

R ^a and R ^b are each independently selected from hydrogen and C ₁ -C ₆ alkyl; or,

^Ra and ^Rb together with the nitrogen atom to which they are attached form a 5-6 membered nitrogen-containing heterocyclic group, wherein the 5-6 membered nitrogen-containing heterocyclic group is optionally substituted with one or more groups selected from halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, carboxyl, ester, _{C1 - C6} alkyl, _C1 - _C6 alkoxy, C1- _C6 haloalkyl, _C1 - _C6 haloalkoxy;

s is 0, 1, or 2;

R ² , Z and n are as defined in the general formula (I).

In another preferred embodiment, the compound of general formula (I) according to the present invention or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, is a compound of general formula (VI) or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof,

in:

Ring A is selected from C ₃ -C ₁₂ cycloalkyl and 3 to 14-membered heterocyclyl, preferably 3 to 14-membered heterocyclyl, more preferably 3 to 6-membered monocyclic heterocyclyl, 6 to 11-membered bicyclic heterocyclyl, further preferably 6 to 10-membered fused heterocyclyl, 6 to 10-membered spiro heterocyclyl;

R ^d is selected from hydrogen, halogen, amino, cyano, oxo, hydroxyl, mercapto, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkoxy;

Each ^R3 is independently selected from hydrogen, halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, _C1 - _C6 alkyl, _C1 - _C6 alkoxy, _C2 - _C6 alkenyl, C2- _C6 alkynyl, _C3 - _C6 cycloalkyl, ₃ to 6 membered heterocyclyl, _C6 - _C10 aryl, 5 to 10 membered heteroaryl, the _C1 - _C6 alkyl, _C1 - _C6 alkoxy, _C2 - _C6 alkenyl, _C2 - _C6 alkynyl, _C3 - _C6 cycloalkyl, 3 to 6 membered heterocyclyl, _C6 - _C10 aryl, 5 to 10 membered heteroaryl being optionally selected from halogen, amino, nitro, cyano, hydroxyl, thiol, carboxyl, ester, oxo, _C1 - _C6 alkyl, _C1 - _C6 alkoxy, _C1 - _C6 haloalkyl, _C1 -C ₆ haloalkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclyl, C ₆ -C ₁₀ aryl, 5 to 10 membered heteroaryl, or one or more groups; preferably R ³ is hydrogen;

r is 0, 1, 2, 3 or 4, preferably 0, 1 or 2, more preferably 0 or 1;

Each R ⁴ is independently selected from halogen, cyano, hydroxyl, mercapto, C ₁ -C ₆ alkyl, -NR ^a R ^b , -OR ^a , wherein the C ₁ -C ₆ alkyl is optionally substituted with halogen; preferably R ⁴ is selected from halogen;

R ^a and R ^b are each independently selected from hydrogen and C ₁ -C ₆ alkyl; or,

^Ra and ^Rb together with the nitrogen atom to which they are attached form a 5-6 membered nitrogen-containing heterocyclic group, wherein the 5-6 membered nitrogen-containing heterocyclic group is optionally substituted with one or more groups selected from halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, carboxyl, ester, _{C1 - C6} alkyl, _C1 - _C6 alkoxy, C1- _C6 haloalkyl, _C1 - _C6 haloalkoxy;

s is 0, 1, or 2;

R ² , Z and n are as defined in the general formula (I).

In a specific embodiment, according to the compounds represented by general formula (III-1), general formula (III-2), general formula (III-3), general formula (IIIA), general formula (IIIB), general formula (IIIC), general formula (IIID), general formula (IIIE), general formula (IIIF), general formula (IIID-1), general formula (IIIE-1), general formula (IIIF-1) or their stereoisomers, tautomers, mesoforms, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, wherein X ₁ is selected from -CH ₂ -, -O-, and -NH-.

In another specific embodiment, according to the compounds represented by general formula (III-1), general formula (III-2), general formula (III-3), general formula (IIIA), general formula (IIIB), general formula (IIIC), general formula (IIID), general formula (IIIE), general formula (IIIF), general formula (IIID-1), general formula (IIIE-1), general formula (IIIF-1) or their stereoisomers, tautomers, mesoforms, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, wherein X ₂ is selected from -CH ₂ -.

In another specific embodiment, according to the compounds represented by general formula (III-1), general formula (III-2), general formula (III-3), general formula (IIIA), general formula (IIIB), general formula (IIIC), general formula (IIID), general formula (IIIE), general formula (IIIF), general formula (IIID-1), general formula (IIIE-1), general formula (IIIF-1) or their stereoisomers, tautomers, mesoforms, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, wherein _X1 and _X2 are both _-CH2- .

In another specific embodiment, according to the compounds represented by general formula (III-1), general formula (III-2), general formula (III-3), general formula (IIIA), general formula (IIIB), general formula (IIIC), general formula (IIID), general formula (IIIE), general formula (IIIF), general formula (IIID-1), general formula (IIIE-1), general formula (IIIF-1) or their stereoisomers, tautomers, mesoforms, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, wherein _n1 is 0 or 1.

In another specific embodiment, according to the compounds represented by general formula (III-1), general formula (III-2), general formula (III-3), general formula (IIIA), general formula (IIIB), general formula (IIIC), general formula (IIID), general formula (IIIE), general formula (IIIF), general formula (IIID-1), general formula (IIIE-1), general formula (IIIF-1) or their stereoisomers, tautomers, racemates, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, wherein _n2 is 0, 1 or 2.

In another specific embodiment, according to the compounds of the general formula (III-1), general formula (III-2), general formula (III-3), general formula (IIIA), general formula (IIIB), general formula (IIIC), general formula (IIID), general formula (IIIE), general formula (IIIF), general formula (IIID-1), general formula (IIIE-1), general formula (IIIF-1) or their stereoisomers, tautomers, mesoforms, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, wherein R ^c is hydrogen and q is 1.

In another specific embodiment, according to the compounds represented by general formula (III-1), general formula (III-2), general formula (III-3), general formula (IIIA), general formula (IIIB), general formula (IIIC), general formula (IIID), general formula (IIIE), general formula (IIIF), general formula (IIID-1), general formula (IIIE-1), general formula (IIIF-1) or their stereoisomers, tautomers, racemates, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, wherein q is 0.

In another specific embodiment, according to the compound represented by general formula (IIIA), general formula (IIIB), general formula (IIIC), general formula (IIID), general formula (IIIE), general formula (IIIF), general formula (IIID-1), general formula (IIIE-1), general formula (IIIF-1), general formula (VIA), general formula (VIB), general formula (VIC), general formula (VID), general formula (IVC-1), general formula (IVD-1), general formula (VV) or general formula (VI) or its stereoisomers, tautomers, mesoforms, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, wherein R ³ is hydrogen and r is 1.

In another specific embodiment, according to the compound represented by general formula (IIIA), general formula (IIIB), general formula (IIIC), general formula (IIID), general formula (IIIE), general formula (IIIF), general formula (IIID-1), general formula (IIIE-1), general formula (IIIF-1), general formula (VIA), general formula (VIB), general formula (VIC), general formula (VID), general formula (IVC-1), general formula (IVD-1), general formula (VV) or general formula (VI) or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein r is 0.

In another specific embodiment, according to the present invention, the compound represented by general formula (IIID), general formula (IIIE), general formula (IIIF) or general formula (IIID-1), general formula (IIIE-1), general formula (IIIF-1), general formula (VIC), general formula (VID), general formula (IVC-1), general formula (IVD-1), general formula (VV) or general formula (VI) or its stereoisomers, tautomers, racemates, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, wherein R ⁴ is hydrogen or halogen, preferably halogen, and s is 1.

In a preferred embodiment, the compound of the general formula according to the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein Z is CH.

In another preferred embodiment, the compound of the general formula according to the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein Z is N.

In another preferred embodiment, the compound of the general formula according to the present invention or its stereoisomers, tautomers, mesomers, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, wherein each R ² is independently selected from hydrogen, halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C 2 -C 6 alkenyl, C ₂ -C ₆ alkynyl, and the C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₂ -C ₆ alkenyl, C _{2 -C 6 alkynyl} is optionally substituted with one or more groups selected from halogen, amino, nitro, cyano, hydroxyl, thiol, carboxyl, ester, oxo, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkoxy;

n is 0, 1 or 2, preferably 0 or 1, more preferably 0.

Typical compounds of the present invention include but are not limited to the following compounds:

or a stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof.

The present invention further provides a method for preparing a compound represented by general formula (IIIA) or general formula (IIIB) or general formula (IIIC) or its stereoisomers, tautomers, mesomorphs, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, comprising the following steps:

The compound of formula IIIAa and the compound of formula IIIAc undergo coupling reaction under alkaline conditions in the presence of a catalyst to obtain a compound of formula IIIAb; the compound of formula IIIAb is deprotected under acidic conditions to obtain a compound represented by general formula (IIIA);

The compound of formula IIIAa and the compound of formula IIIBc undergo coupling reaction under alkaline conditions in the presence of a catalyst to obtain a compound of formula IIIBb; the compound of formula IIIBb is deprotected under acidic conditions to obtain a compound represented by general formula (IIIB);

The compound of formula IIIAa and the compound of formula IIICc undergo coupling reaction under alkaline conditions in the presence of a catalyst to obtain a compound of formula IIICb; the compound of formula IIICb is deprotected under acidic conditions to obtain a compound represented by general formula (IIIC);

in:

The catalyst is preferably benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate;

The alkaline condition is preferably N,N-diisopropylethylamine;

The acidic condition is preferably hydrochloric acid;

^Rx is an amino protecting group, preferably Boc;

G, Z, R ² , R ³ , X ₁ , X ₂ , R ^c , n, n ₁ , n ₂ , q and r are as defined in Formula (IIIA), (IIIB) or (IIIC).

The present invention further relates to a method for preparing a compound represented by general formula (IIID) or general formula (IIIE) or general formula (IIIF) or its stereoisomers, tautomers, mesomorphs, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, comprising the following steps:

The compound of formula IIIAa and the compound of formula IIIDc undergo coupling reaction under alkaline conditions in the presence of a catalyst to obtain a compound of formula IIIDb; the compound of formula IIIDb is deprotected under acidic conditions to obtain a compound represented by general formula (IIID);

The compound of formula IIIAa and the compound of formula IIIEc undergo coupling reaction under alkaline conditions in the presence of a catalyst to obtain a compound of formula IIIEb; the compound of formula IIIEb is deprotected under acidic conditions to obtain a compound represented by general formula (IIIE);

The compound of formula IIIAa and the compound of formula IIIFc undergo coupling reaction under alkaline conditions in the presence of a catalyst to obtain a compound of formula IIIFb; the compound of formula IIIFb is deprotected under acidic conditions to obtain a compound represented by general formula (IIIF);

in:

The catalyst is preferably benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate;

The alkaline condition is preferably N,N-diisopropylethylamine;

The acidic condition is preferably hydrochloric acid;

^Rx is an amino protecting group, preferably Boc;

Z, R ² , R ³ , R ⁴ , X ₁ , X ₂ , R ^c , n, n ₁ , n ₂ , q, r and s are as defined in formula (IIID), (IIIE) or (IIIF).

The present invention further relates to a method for preparing a compound represented by general formula (IIID-1) or general formula (IIIE-1) or general formula (IIIF-1) or its stereoisomers, tautomers, mesomorphs, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, comprising the following steps:

The compound of formula IIIAa and the compound of formula IIID-1c undergo coupling reaction under alkaline conditions in the presence of a catalyst to obtain a compound of formula IIID-1b; the compound of formula IIID-1b is deprotected under acidic conditions to obtain a compound represented by general formula (IIID-1);

The compound of formula IIIAa and the compound of formula IIIE-1c undergo coupling reaction under alkaline conditions in the presence of a catalyst to obtain a compound of formula IIIE-1b; the compound of formula IIIE-1b is deprotected under acidic conditions to obtain a compound represented by general formula (IIIE-1);

The compound of formula IIIAa and the compound of formula IIIF-1c undergo coupling reaction under alkaline conditions in the presence of a catalyst to obtain a compound of formula IIIF-1b; the compound of formula IIIF-1b is deprotected under acidic conditions to obtain a compound represented by general formula (IIIF-1);

in:

The catalyst is preferably benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate;

The alkaline condition is preferably N,N-diisopropylethylamine;

The acidic condition is preferably hydrochloric acid;

^Rx is an amino protecting group, preferably Boc;

Z, R ² , R ³ , R ⁴ , X ₁ , X ₂ , R ^c , n, n ₁ , n ₂ , q, r and s are as defined in general formula (IIID-1), general formula (IIIE-1) or general formula (IIIF-1).

The present invention further relates to a method for preparing a compound represented by general formula (IVA) or its stereoisomers, tautomers, mesomers, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, comprising the following steps:

The compound of formula IIIAa and the compound of formula IVAc undergo coupling reaction under alkaline conditions in the presence of a catalyst to obtain a compound of formula IVAb; the compound of formula IVAb is deprotected under acidic conditions to obtain a compound represented by general formula (IVA);

The catalyst is preferably benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate;

The alkaline condition is preferably N,N-diisopropylethylamine;

The acidic condition is preferably hydrochloric acid;

in:

^Rx is an amino protecting group, preferably Boc;

Ring E ₁ , G, Z, R ² , R ³ , ^Re , n, r and t are as defined in the general formula (IVA).

The present invention further relates to a method for preparing a compound represented by general formula (IVB) or its stereoisomers, tautomers, mesomers, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, comprising the following steps:

The compound of formula IIIAa and the compound of formula IVBc undergo coupling reaction under alkaline conditions in the presence of a catalyst to obtain a compound of formula IVBb; the compound of formula IVBb is deprotected under acidic conditions to obtain a compound represented by general formula (IVB);

The catalyst is preferably benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate;

The alkaline condition is preferably N,N-diisopropylethylamine;

The acidic condition is preferably hydrochloric acid;

in:

^Rx is an amino protecting group, preferably Boc;

Ring E ₂ , G, Z, R ² , R ³ , ^Re , n, r, and t are as defined in the general formula (IVB).

The present invention further relates to a method for preparing a compound represented by general formula (IVC) or its stereoisomers, tautomers, mesomers, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, comprising the following steps:

The compound of formula IIIAa and the compound of formula IVCc undergo coupling reaction under alkaline conditions in the presence of a catalyst to obtain a compound of formula IVCb; the compound of formula IVCb is deprotected under acidic conditions to obtain a compound represented by general formula (IVC);

The catalyst is preferably benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate;

The alkaline condition is preferably N,N-diisopropylethylamine;

The acidic condition is preferably hydrochloric acid;

in:

^Rx is an amino protecting group, preferably Boc;

Ring E ₁ , Z, R ² , R ³ , R ⁴ , ^Re , n, r, s and t are as defined in the general formula (IVC).

The present invention further relates to a method for preparing a compound represented by general formula (IVD) or its stereoisomers, tautomers, mesomers, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, comprising the following steps:

The compound of formula IIIAa and the compound of formula IVDc undergo coupling reaction under alkaline conditions in the presence of a catalyst to obtain a compound of formula IVDb; the compound of formula IVDb is deprotected under acidic conditions to obtain a compound represented by general formula (IVD);

The catalyst is preferably benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate;

The alkaline condition is preferably N,N-diisopropylethylamine;

The acidic condition is preferably hydrochloric acid;

in:

^Rx is an amino protecting group, preferably Boc;

Ring E ₂ , Z, R ² , R ³ , R ⁴ , ^Re , n, r, s and t are as defined in the general formula (IVD).

The present invention further relates to a method for preparing a compound represented by general formula (IVC-1) or its stereoisomers, tautomers, mesomers, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, comprising the following steps:

The compound of formula IIIAa and the compound of formula IVC-1c undergo coupling reaction under alkaline conditions in the presence of a catalyst to obtain a compound of formula IVC-1b; the compound of formula IVC-1b is deprotected under acidic conditions to obtain a compound represented by general formula (IVC-1);

The catalyst is preferably benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate;

The alkaline condition is preferably N,N-diisopropylethylamine;

The acidic condition is preferably hydrochloric acid;

in:

^Rx is an amino protecting group, preferably Boc;

Ring E ₁ , Z, R ² , R ³ , R ⁴ , ^Re , n, r, s and t are as defined in the general formula (IVC-1).

The present invention further relates to a method for preparing a compound represented by general formula (IVD-1) or its stereoisomers, tautomers, mesomers, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, comprising the following steps:

The compound of formula IIIAa and the compound of formula IVD-1c undergo coupling reaction under alkaline conditions in the presence of a catalyst to obtain a compound of formula IVD-1b; the compound of formula IVD-1b is deprotected under acidic conditions to obtain a compound represented by general formula (IVD-1);

The catalyst is preferably benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate;

The alkaline condition is preferably N,N-diisopropylethylamine;

The acidic condition is preferably hydrochloric acid;

in:

^Rx is an amino protecting group, preferably Boc;

Ring E ₂ , Z, R ² , R ³ , R ⁴ , ^Re , n, r, s and t are as defined in the general formula (IVD-1).

The present invention also provides a pharmaceutical composition, which comprises the general formula compound according to the present invention or its stereoisomers, tautomers, mesomorphs, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, and one or more pharmaceutically acceptable carriers, diluents or excipients.

The present invention also relates to the use of the general formula compound according to the present invention or its stereoisomers, tautomers, mesomers, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions containing the same in the preparation of drugs for inhibiting AKT1/2/3.

The present invention also relates to the use of the general formula compound according to the present invention or its stereoisomers, tautomers, mesomers, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions containing the same, in the preparation of drugs for preventing and/or treating diseases associated with AKT1/2/3 kinase activity, wherein the disease is preferably cancer, in particular cancer characterized by amplification or overexpression of AKT1/2/3, wherein the cancer is preferably ovarian cancer, breast cancer, Prostate cancer, glioma, glioma, gastric cancer, fallopian tube cancer, lung cancer, peritoneal tumors, melanoma, brain cancer, esophageal cancer, liver cancer, pancreatic cancer, colorectal cancer, lung cancer, kidney cancer, cervical cancer, skin cancer, neuroblastoma, sarcoma, bone cancer, uterine cancer, endometrial cancer, head and neck tumors, multiple myeloma, lymphoma, non-Hodgkin's lymphoma, non-small cell lung cancer, polycythemia vera, leukemia, thyroid tumors, bladder cancer and gallbladder cancer, particularly preferably ovarian cancer, breast cancer and prostate cancer.

The present invention also relates to the general formula compound according to the present invention or its stereoisomer, tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt or pharmaceutical composition containing the same, which is used as an AKT1/2/3 inhibitor.

The present invention also relates to the general formula compound according to the present invention or its stereoisomers, tautomers, mesomers, racemates, enantiomers, diastereomers, or mixtures thereof, or prodrugs thereof, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions comprising the same, for use in treating and/or preventing diseases associated with AKT1/2/3 kinase activity, wherein the disease is preferably cancer, in particular cancers characterized by amplification or overexpression of AKT1/2/3, wherein the cancer is preferably ovarian cancer, breast cancer, prostate cancer, Preferably, the present invention is directed to prostate cancer, glioma, glioma, gastric cancer, fallopian tube cancer, lung cancer, peritoneal tumor, melanoma, brain cancer, esophageal cancer, liver cancer, pancreatic cancer, colorectal cancer, lung cancer, kidney cancer, cervical cancer, skin cancer, neuroblastoma, sarcoma, bone cancer, uterine cancer, endometrial cancer, head and neck tumor, multiple myeloma, lymphoma, non-Hodgkin's lymphoma, non-small cell lung cancer, polycythemia vera, leukemia, thyroid tumor, bladder cancer and gallbladder cancer, particularly preferably ovarian cancer, breast cancer and prostate cancer.

The present invention also relates to the general formula compound according to the present invention or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt or pharmaceutical composition comprising the same, which is used as a drug, especially a drug for preventing and/or treating diseases related to AKT1/2/3 kinase activity, wherein the disease is preferably cancer, especially cancer characterized by amplification or overexpression of AKT1/2/3, wherein the cancer is preferably ovarian cancer, breast cancer, or Preferably, the present invention is directed to cancer, prostate cancer, glioma, glioma, gastric cancer, fallopian tube cancer, lung cancer, peritoneal tumor, melanoma, brain cancer, esophageal cancer, liver cancer, pancreatic cancer, colorectal cancer, lung cancer, kidney cancer, cervical cancer, skin cancer, neuroblastoma, sarcoma, bone cancer, uterine cancer, endometrial cancer, head and neck tumor, multiple myeloma, lymphoma, non-Hodgkin's lymphoma, non-small cell lung cancer, polycythemia vera, leukemia, thyroid tumor, bladder cancer and gallbladder cancer, particularly preferably ovarian cancer, breast cancer and prostate cancer.

The present invention also relates to a method for inhibiting AKT1/2/3, which comprises administering to a subject in need thereof an effective amount of a compound of the general formula according to the present invention or its stereoisomers, tautomers, mesomers, racemates, enantiomers, diastereomers, or mixtures thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same.

The present invention also relates to a method for inhibiting cancer cell proliferation, inhibiting cancer cell invasion or inducing cancer cell apoptosis, which comprises administering to a subject in need thereof an effective amount of the general formula compound according to the present invention or its stereoisomers, tautomers, mesomorphs, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions comprising the same.

The present invention also relates to a method for preventing and/or treating a disease associated with AKT1/2/3 kinase activity, comprising administering to a subject in need thereof an effective amount of a compound of the general formula according to the present invention or its stereoisomer, tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, wherein the disease is preferably cancer, in particular cancer characterized by amplification or overexpression of AKT1/2/3, wherein the cancer is preferably ovarian cancer. Preferably, the present invention is directed to cancer, breast cancer, prostate cancer, glioma, glioma, gastric cancer, fallopian tube cancer, lung cancer, peritoneal tumor, melanoma, brain cancer, esophageal cancer, liver cancer, pancreatic cancer, colorectal cancer, lung cancer, kidney cancer, cervical cancer, skin cancer, neuroblastoma, sarcoma, bone cancer, uterine cancer, endometrial cancer, head and neck tumor, multiple myeloma, lymphoma, non-Hodgkin's lymphoma, non-small cell lung cancer, polycythemia vera, leukemia, thyroid tumor, bladder cancer and gallbladder cancer, particularly preferably ovarian cancer, breast cancer and prostate cancer.

The compound according to the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt or pharmaceutical composition containing the same can be administered simultaneously, separately or sequentially with another anticancer therapeutic agent or anticancer treatment method.

Detailed description of the invention

Unless stated otherwise, the terms used in the specification and claims have the following meanings.

"Alkyl" refers to a saturated aliphatic hydrocarbon group, including straight and branched chain groups having the specified number of carbon atoms. Alkyl groups generally contain 1 to 20 carbon atoms ( _C1 - _C20 alkyl), preferably 1 to 12 carbon atoms ( _C1 - _C12 alkyl), more preferably 1 to 8 carbon atoms ( _C1 - _C8 alkyl) or 1 to 6 carbon atoms ( _C1 - _C6 alkyl) or 1 to 4 carbon atoms ( _C1 - _C4 alkyl). Non-limiting examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2 ,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched chain isomers thereof, etc. The alkyl group may be substituted or unsubstituted, and when substituted, the substituents may be substituted at any available point of attachment. Optional substituents suitable for alkyl include, but are not limited to, halogen, amino, nitro, cyano (-CN), hydroxyl (-OH), thiol (-SH), carboxyl (-COOH), ester, oxo (=O), thio (=S), =N-CN, alkoxy, aryloxy, heteroaryloxy, haloalkyl, haloalkoxy, hydroxyalkyl, alkylamino, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl. In some embodiments, alkyl is optionally substituted with one or more substituents, preferably 1-3 substituents.

The term "alkoxy" refers to -O-(alkyl), wherein alkyl is defined as described above. Non-limiting examples of alkoxy include: methoxy, ethoxy, propoxy, butoxy. Alkoxy can be optionally substituted or unsubstituted, and when substituted, substituents are preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, alkylthio, alkylamino, halogen, sulfhydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocyclic radical, aryl, heteroaryl, cycloalkyloxy, heterocyclic radical oxygen base, cycloalkylthio, heterocyclic radical thio, carboxyl or carboxylate.

The term "alkenyl" refers to an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon double bond. Alkenyl groups typically contain 2 to 20 carbon atoms ( _C2 - _C20 alkenyl), preferably 2 to 12 carbon atoms ( _C2 - _C12 alkenyl), more preferably 2 to 8 carbon atoms ( _C2 - _C8 alkenyl) or 2 to 6 carbon atoms ( _C2 - _C6 alkenyl) or 2 to 4 carbon atoms ( _C2 - _C4 alkenyl). Non-limiting examples of alkenyl groups include vinyl, 1-propenyl, 2-propenyl, 1-, 2- or 3-butenyl, and the like. The alkenyl group may be substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkyloxy, heterocyclyloxy, cycloalkylthio, heterocyclylthio.

The term "alkynyl" refers to an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon triple bond. Alkynyl groups typically contain 2-20 carbon atoms ( _C2 - _C20 alkynyl), preferably 2 to 12 carbon atoms ( _C2 - _C12 alkynyl), more preferably 2 to 8 carbon atoms ( _C2 - _C8 alkynyl) or 2 to 6 carbon atoms ( _C2 - _C6 alkynyl) or 2 to 4 carbon atoms ( _C2 - _C4 alkynyl). Non-limiting examples of alkynyl groups include ethynyl, propynyl, butynyl, etc. Alkynyl groups may be substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfhydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkyloxy, heterocyclyloxy, cycloalkylthio, heterocyclylthio.

The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, and the cycloalkyl ring comprises 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, more preferably 3 to 6 carbon atoms. The limiting examples of monocyclic cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl etc. Polycyclic cycloalkyl includes the cycloalkyl of spiro ring, condensed ring and bridge ring.

The term "spirocycloalkyl" refers to a polycyclic group that shares a carbon atom (called a spiral atom) between 5 to 20 monocyclic rings, which may contain one or more double bonds, but no ring has a completely conjugated π electron system. Preferably, it is 6 to 14 yuan, such as 6 to 10 yuan, 6 to 11 yuan or 7 to 10 yuan. According to the number of spiral atoms shared between rings, the spirocycloalkyl is divided into a single spiral cycloalkyl, a double spiral cycloalkyl or a multi-spirocycloalkyl, preferably a single spiral cycloalkyl and a double spiral cycloalkyl. More preferably, it is 4 yuan/4 yuan, 4 yuan/5 yuan, 4 yuan/6 yuan, 5 yuan/5 yuan or 5 yuan/6 yuan single spiral cycloalkyl. Non-limiting examples of spirocycloalkyl include:

The term "condensed cycloalkyl" refers to a 5 to 20-membered, all-carbon polycyclic group in which each ring in the system shares a pair of adjacent carbon atoms with other rings in the system, wherein one or more rings may contain one or more double bonds, but no ring has a completely conjugated π electron system. Preferably, it is 6 to 14 yuan, such as 6 to 10 yuan, 6 to 11 yuan or 7 to 10 yuan. According to the number of constituent rings, it can be divided into a bicyclic, tricyclic, tetracyclic or polycyclic condensed cycloalkyl, preferably a bicyclic or tricyclic, more preferably a 5-yuan/5-yuan or 5-yuan/6-yuan bicyclic alkyl. Non-limiting examples of condensed cycloalkyls include:

The term "bridged cycloalkyl" refers to a 5 to 20-membered, all-carbon polycyclic group in which any two rings share two carbon atoms that are not directly connected, which may contain one or more double bonds, but no ring has a completely conjugated π electron system. Preferably, it is 6 to 14 members, and more preferably 7 to 10 members. According to the number of constituent rings, it can be divided into a bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyl, preferably a bicyclic, tricyclic or tetracyclic, and more preferably a bicyclic or tricyclic. Non-limiting examples of bridged cycloalkyl include:

The cycloalkyl ring may be fused to an aryl, heteroaryl or heterocyclyl ring, wherein the ring attached to the parent structure is a cycloalkyl, non-limiting examples of which include indanyl, tetrahydronaphthyl, benzocycloheptanyl, etc. The cycloalkyl may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkyloxy, heterocyclyloxy, cycloalkylthio, heterocyclylthio, oxo, carboxyl or carboxylate.

The term "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent containing 3 to 20 ring atoms, one or more of which is a heteroatom selected from nitrogen, oxygen or S(O) _m (wherein m is an integer from 0 to 2), but excluding the ring part of -OO-, -OS- or -SS-, and the remaining ring atoms are carbon. Preferably, it contains 3 to 14 ring atoms, of which 1 to 4 are heteroatoms; or 3 to 8 ring atoms, of which 1 to 3 are heteroatoms; or 5 to 7 ring atoms, of which 1 to 2 or 1 to 3 are heteroatoms; or 3 to 8 ring atoms, of which 1 to 2 are heteroatoms. Non-limiting examples of monocyclic heterocyclyls include oxetanyl, azetidinyl, pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, tetrahydrothienyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, pyranyl, etc. Polycyclic heterocyclyls include spiro, fused and bridged heterocyclyls.

The term "spiro heterocyclic group" refers to a polycyclic heterocyclic group in which one atom (called a spiral atom) is shared between 5 to 20 monocyclic rings, wherein one or more ring atoms are heteroatoms selected from nitrogen, oxygen or S(O) _m (wherein m is an integer from 0 to 2), and the remaining ring atoms are carbon. It may contain one or more double bonds, but no ring has a completely conjugated π electron system. It is preferably 6 to 14 yuan, for example 6 to 10 yuan, 6 to 11 yuan or 7 to 10 yuan. According to the number of shared spiral atoms between rings, the spiral heterocyclic group is divided into a single spiral heterocyclic group, a double spiral heterocyclic group or a multi-spiro heterocyclic group, preferably a single spiral heterocyclic group and a double spiral heterocyclic group. More preferably, it is a 4-yuan/4-yuan, 4-yuan/5-yuan, 4-yuan/6-yuan, 5-yuan/5-yuan or 5-yuan/6-yuan single spiral heterocyclic group. Non-limiting examples of spiral heterocyclic groups include:

The term "fused heterocyclic group" refers to a polycyclic heterocyclic group of 5 to 20 members, each ring in the system shares a pair of atoms adjacent to other rings in the system, one or more rings may contain one or more double bonds, but no ring has a completely conjugated π electron system, wherein one or more ring atoms are heteroatoms selected from nitrogen, oxygen or S(O) _m (wherein m is an integer from 0 to 2), and the remaining ring atoms are carbon. Preferably, it is 6 to 14 members, such as 6 to 10 members, 6 to 11 members or 7 to 10 members. According to the number of constituent rings, it can be divided into a bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic group, preferably a bicyclic or tricyclic group, more preferably a 5-membered/5-membered or 5-membered/6-membered bicyclic fused heterocyclic group. Non-limiting examples of fused heterocyclic groups include:

The term "bridged heterocyclic group" refers to a polycyclic heterocyclic group of 5 to 14 members, in which any two rings share two atoms that are not directly connected, which may contain one or more double bonds, but none of the rings has a completely conjugated π electron system, wherein one or more ring atoms are heteroatoms selected from nitrogen, oxygen or S(O) _m (wherein m is an integer from 0 to 2), and the remaining ring atoms are carbon. Preferably, it is 6 to 14 members, more preferably 6 to 10 members. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged heterocyclic groups, preferably bicyclic, tricyclic or tetracyclic, and more preferably bicyclic or tricyclic. Non-limiting examples of bridged heterocyclic groups include:

The heterocyclyl ring may be fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring attached to the parent structure is a heterocyclyl, non-limiting examples of which include:

The heterocyclyl group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkyloxy, heterocyclyloxy, cycloalkylthio, heterocyclylthio, oxo, carboxyl or carboxylate.

The term "aryl" refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (i.e., rings that share adjacent pairs of carbon atoms) group having a conjugated π electron system, preferably 6- to 10-membered, such as phenyl and naphthyl, more preferably phenyl. The aryl ring may be fused to a heteroaryl, heterocyclyl or cycloalkyl ring, wherein the ring connected to the parent structure is an aryl ring, non-limiting examples of which include:

The aryl group may be substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkyloxy, heterocyclyloxy, cycloalkylthio, heterocyclylthio, carboxyl or carboxylate.

The term "heteroaryl" refers to a heteroaromatic system containing 1 to 4 heteroatoms and 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen. The heteroaryl group is preferably 5 to 10 members, containing 1 to 3 heteroatoms; more preferably 5 or 6 members, containing 1 to 2 heteroatoms; for example, imidazolyl, furanyl, thienyl, thiazolyl, pyrazolyl, oxazolyl, pyrrolyl, tetrazolyl, pyridyl, pyrimidinyl, thiadiazole, pyrazinyl, etc. The heteroaryl ring can be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring, non-limiting examples of which include:

The heteroaryl group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkyloxy, heterocyclyloxy, cycloalkylthio, heterocyclylthio, carboxyl or carboxylate.

The term "haloalkyl" refers to an alkyl group substituted with one or more halogens, wherein alkyl is as defined above.

The term "haloalkoxy" refers to an alkoxy group substituted with one or more halogens, wherein alkoxy is as defined above.

The term "alkylthio" refers to -S-(alkyl) wherein alkyl is as defined above.

The term "alkylamino" refers to -NH-(alkyl), -N(alkyl) _2- , -NH-(cycloalkyl) or -N-(cycloalkyl) ₂ , wherein alkyl and cycloalkyl are as defined above.

The term "cycloalkyloxy" refers to -O-(cycloalkyl), wherein cycloalkyl is as defined above. Non-limiting examples of cycloalkyloxy include: cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy.

The term "cycloalkylthio" refers to -S-(cycloalkyl) wherein cycloalkyl is as previously defined.

The term "heterocyclyloxy" refers to -O-(heterocyclyl) wherein heterocyclyl is as defined above.

The term "heterocyclylthio" refers to -S-(heterocyclyl) wherein heterocyclyl is as previously described.

The term "hydroxy" refers to an -OH group.

The term "halogen" refers to fluorine, chlorine, bromine or iodine.

The term "amino" refers to _-NH2 .

The term "cyano" refers to -CN.

The term "nitro" refers to _-NO2 .

The term "oxo" refers to =0.

The term "carboxy" refers to -C(O)OH.

The term "thiol" refers to -SH.

The term "ester" or "carboxylate" refers to -C(O)O(alkyl) or -C(O)O(cycloalkyl) wherein alkyl and cycloalkyl are as defined above.

The term "acyl" refers to a compound containing a -C(O)R group, wherein R is alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl.

The term "amino protecting group" is to protect the amino group with a group that is easily removed in order to keep the amino group unchanged when other parts of the molecule react. Non-limiting examples include tert-butyloxycarbonyl (Boc), acetyl, benzyl, allyl and p-methoxybenzyl. These groups can be optionally substituted with 1-3 substituents selected from halogen, alkoxy or nitro. The amino protecting group is preferably tert-butyloxycarbonyl.

"Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and the description includes instances where the event or circumstance occurs and instances where it does not occur. For example, "heterocyclyl optionally substituted with alkyl" means that alkyl may but need not be present, and the description includes instances where the heterocyclyl is substituted with alkyl and instances where the heterocyclyl is not substituted with alkyl.

"Substituted" means that one or more hydrogen atoms, preferably up to 5, more preferably 1 to 3 hydrogen atoms in the group are replaced independently of each other by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and the skilled person can determine (by experiment or theory) possible or impossible substitutions without undue effort. For example, amino or hydroxy groups with free hydrogens may be unstable when combined with carbon atoms with unsaturated (e.g. olefinic) bonds.

"Pharmaceutical composition" means a mixture containing one or more compounds described herein or their physiologically/pharmaceutically acceptable salts and other chemical components, as well as other components such as physiologically/pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitate the absorption of the active ingredient, and thus exert biological activity.

The term "pharmaceutically acceptable salt" and "pharmaceutically acceptable salt" are used interchangeably and refer to salts of the compounds of the present invention that are safe and effective when used in mammals and have the desired biological activity.

"Cancer" refers to any malignant and/or invasive growth or tumor (caused by abnormal cell growth). Cancer includes solid tumors named after the cell types that form them, cancers of the blood, bone marrow or lymphatic system. Examples of solid tumors include sarcomas and carcinomas. Hematological cancers include, but are not limited to, leukemias, lymphomas and myelomas. Cancer also includes primary cancers originating from specific parts of the body, metastatic cancers that have spread from the site where it started to other parts of the body, recurrences from the original primary cancer after relief, and second primary cancers (this is a new primary cancer in a person with a previous cancer history that is different from the new primary cancer). In some embodiments provided herein, cancer can be selected from breast cancer, ovarian cancer, prostate cancer, bladder cancer, uterine cancer, lung cancer, esophageal cancer, liver cancer, pancreatic cancer and gastric cancer. In some such embodiments, cancer is characterized by amplification or overexpression of AKT1/2/3.

Stereoisomers described herein may include cis and trans isomers, optical isomers such as (R) and (S) enantiomers, diastereomers, geometric isomers, rotational isomers, atropisomers, conformational isomers and tautomers of the compounds of the invention (including compounds exhibiting more than one isomeric type); and mixtures thereof (e.g., racemates and diastereomeric pairs).

The compounds of the present invention can exhibit tautomerism and structural isomerism. For example, the compound can exist in several tautomeric forms, including enol and imine forms and ketone and enamine forms, and geometric isomers and mixtures thereof. All such tautomeric forms are included within the scope of the compounds of the present invention. Tautomers exist as mixtures of tautomeric groups in solution. In solid forms, usually one tautomer is dominant. Even if one tautomer can be described, the present invention includes all tautomers of the compounds provided.

Conventional techniques for the preparation/isolation of individual enantiomers include chiral synthesis from suitable optically pure precursors or resolution of the racemate (or the racemate of a salt or derivative) using, for example, chiral high pressure liquid chromatography (HPLC) or supercritical fluid chromatography (SFC).

The enantiomeric purity of the compounds described herein can be described in terms of enantiomeric excess (ee), which indicates the extent to which a sample contains one enantiomer in greater amounts than the other enantiomer. A racemic mixture has an ee of 0%, while a single, completely pure enantiomer has an ee of 100%. Similarly, diastereomeric purity can be described in terms of diastereomeric excess (de).

According to the conventional methods in the field of the present invention, the compounds of the present invention can form pharmaceutically acceptable basic addition salts or acid addition salts with bases or acids. The bases include inorganic bases and organic bases, and acceptable organic bases include diethanolamine, ethanolamine, N-methylglucamine, triethanolamine, tromethamine, etc., and acceptable inorganic bases include aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate and sodium hydroxide, etc. The acid includes inorganic acid and organic acid, and acceptable inorganic acid includes hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrobromic acid, etc. Acceptable organic acids include acetic acid, trifluoroacetic acid, formic acid, ascorbic acid, etc.

The pharmaceutical composition containing the active ingredient may be in a form suitable for oral administration, such as tablets, troches, lozenges, aqueous or oil suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. Oral compositions may be prepared according to any method known in the art for preparing pharmaceutical compositions, and such compositions may contain one or more ingredients selected from the following: sweeteners, flavoring agents, colorants and preservatives to provide pleasing and palatable pharmaceutical preparations. Tablets contain the active ingredient and non-toxic pharmaceutically acceptable excipients suitable for preparing tablets for mixing. These excipients may be inert excipients such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating agents and disintegrants such as microcrystalline cellulose, crosslinked sodium carboxymethyl cellulose, corn starch or alginic acid; binders such as starch, gelatin, polyvinyl pyrrolidone or gum arabic; and lubricants such as magnesium stearate, stearic acid or talc. These tablets may be uncoated or may be coated by known techniques which mask the taste of the drug or delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained release action over a longer period of time. For example, water soluble taste masking materials such as hydroxypropylmethylcellulose or hydroxypropylcellulose, or time extending materials such as ethylcellulose, cellulose acetate butyrate may be used.

Oral preparations may also be provided in hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent such as calcium carbonate, calcium phosphate or kaolin, or in soft gelatin capsules wherein the active ingredient is mixed with a water-soluble carrier such as polyethylene glycol or an oily vehicle such as peanut oil, liquid paraffin or olive oil.

Aqueous suspensions contain the active substance and excipients suitable for preparing aqueous suspensions for mixing. Such excipients are suspending agents, for example, sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone and gum arabic; dispersants or wetting agents, which may be naturally occurring phosphatides such as lecithin, or condensation products of alkylene oxides with fatty acids, for example, polyoxyethylene stearate, or condensation products of ethylene oxide with long chain fatty alcohols, for example, heptadecaethyleneoxy cetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol, for example, polyethylene oxide sorbitan monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example, polyethylene oxide dehydrated sorbitan monooleate. The aqueous suspension may also contain one or more preservatives, for example ethylparaben or n-propylparaben, one or more coloring agents, one or more flavoring agents and one or more sweetening agents, for example sucrose, saccharin or aspartame.

Oil suspensions can be prepared by suspending the active ingredient in a vegetable oil such as peanut oil, olive oil, sesame oil or coconut oil, or a mineral oil such as liquid paraffin. The oil suspension may contain a thickening agent such as beeswax, hard paraffin or cetyl alcohol. The above-mentioned sweeteners and flavoring agents may be added to provide a palatable preparation. These compositions may be preserved by adding an antioxidant such as butylated hydroxyanisole or alpha-tocopherol.

Dispersible powders and granules suitable for preparing aqueous suspensions can provide the active ingredient and a dispersant or wetting agent, a suspending agent or one or more preservatives for mixing by adding water. Suitable dispersants or wetting agents and suspending agents are as described above. Other excipients such as sweeteners, flavoring agents and coloring agents can also be added. These compositions can be preserved by adding antioxidants such as ascorbic acid.

The pharmaceutical composition of the present invention can also be in the form of an oil-in-water emulsion. The oil phase can be a vegetable oil such as olive oil or peanut oil, or a mineral oil such as liquid paraffin or a mixture thereof. Suitable emulsifiers can be naturally occurring phospholipids, such as soybean lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan monooleate, and condensation products of the partial esters and ethylene oxide, such as polyethylene oxide sorbitol monooleate. Emulsions can also contain sweeteners, flavoring agents, preservatives and antioxidants. Syrups and elixirs prepared with sweeteners such as glycerol, propylene glycol, sorbitol or sucrose can be used. Such preparations can also contain a demulcent, a preservative, a coloring agent and an antioxidant.

The pharmaceutical composition of the present invention may be in the form of a sterile injectable aqueous solution. Acceptable vehicles and solvents that may be used are water, Ringer's solution and isotonic sodium chloride solution. The sterile injectable preparation may be a sterile injectable oil-in-water microemulsion in which the active ingredient is dissolved in an oil phase. For example, the active ingredient is dissolved in a mixture of soybean oil and lecithin. The oil solution is then added to a mixture of water and glycerol and processed to form a microemulsion. The injection or microemulsion may be injected into the patient's bloodstream by local mass injection. Alternatively, the solution and microemulsion may be preferably administered in a manner that maintains a constant circulating concentration of the compound of the present invention. To maintain such a constant concentration, a continuous intravenous drug delivery device may be used.

The pharmaceutical composition of the present invention can be in the form of a sterile injection water or oil suspension for intramuscular and subcutaneous administration. The suspension can be prepared according to known techniques with the above-mentioned suitable dispersants or wetting agents and suspending agents. The sterile injection preparation can also be a sterile injection solution or suspension prepared in a non-toxic parenterally acceptable diluent or solvent, such as a solution prepared in 1,3-butanediol. In addition, sterile fixed oils can be conveniently used as solvents or suspension media. For this purpose, any blended fixed oil including synthetic mono- or diglycerides can be used. In addition, fatty acids such as oleic acid can also be used to prepare injections.

It is well known to those skilled in the art that the dosage of a drug depends on a variety of factors, including but not limited to the following factors: the activity of the specific compound used, the patient's age, the patient's weight, the patient's health condition, the patient's behavior, the patient's diet, the administration time, the administration method, the excretion rate, the combination of drugs, etc. In addition, the best treatment method, such as the mode of treatment, the daily dosage of the general compound or the type of pharmaceutically acceptable salt can be verified according to traditional treatment plans.

The present invention may contain the compound shown in the general formula and its pharmaceutically acceptable salt, hydrate or solvate as an active ingredient, mixed with a pharmaceutically acceptable carrier or excipient to prepare a composition, and prepared into a clinically acceptable dosage form. The derivatives of the present invention can be used in combination with other active ingredients, as long as they do not produce other adverse effects, such as allergic reactions. The compounds of the present invention can be used as the only active ingredient, or in combination with other drugs for treating diseases related to AKT kinase activity. Combination therapy is achieved by administering each therapeutic component simultaneously, separately or sequentially.

Synthesis method of the compound of the present invention

In order to achieve the purpose of the present invention, the present invention adopts the following synthesis scheme to prepare the compound of the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof.

The compound represented by the general formula (IIID) of the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt can be prepared by the following scheme 1:

The compound of formula IIIAa and the compound of formula IIIDc undergo coupling reaction under alkaline conditions in the presence of a catalyst to obtain a compound of formula IIIDb; the compound of formula IIIDb is deprotected under acidic conditions to obtain a compound represented by general formula (IIID);

The catalyst is preferably benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate;

The alkaline condition is preferably N,N-diisopropylethylamine;

The acidic condition is preferably hydrochloric acid;

in,

^Rx is an amino protecting group, preferably Boc;

Z, ^R2 , ^R3 , ^R4 , _X1 , _X2 , ^Rc , n, _n1 , _n2 , q, r and s are as defined in the general formula (IIID).

The compound of formula IIIDc can be prepared by the following Scheme 2:

Step 1: In the presence of an activating agent such as pivaloyl chloride, compound a is condensed with a chiral auxiliary such as compound b to obtain compound c;

Step 2: Compound d reacts with a suitable compound such as di-tert-butyl dicarbonate in the presence of a catalyst such as 4-dimethylaminopyridine to obtain an amino-protected compound e;

Step 3: reducing compound e using a reducing agent such as diisobutylaluminum hydride to obtain compound f;

Step 4: In the presence of a catalyst such as p-toluenesulfonic acid, compound f reacts with methanol to obtain compound g;

Step 5: In the presence of a suitable Lewis acid such as titanium tetrachloride and a weak base such as N,N-diisopropylethylamine, compound g is condensed with compound c to obtain compound h. The reaction is preferably carried out at a low temperature such as -78°C to 0°C to obtain acceptable diastereoisomer selectivity in the reaction;

Step 6: Compound h is hydrolyzed in the presence of a base such as lithium hydroxide and a hydrolytic chiral auxiliary such as hydrogen peroxide to give a compound of formula IIIBc.

When Z is a C atom and ^R2 is hydrogen, the compound of formula IIIAa can be prepared by the following scheme 3. When Z is a N atom and ^R2 is selected from other groups, those skilled in the art can easily prepare the compound by referring to this method:

Compound Ij is reacted with Boc protected piperazine to give compound Ik, which is then deprotected to provide compound Il.

DETAILED DESCRIPTION

The present invention is further described by examples, but it is to be understood that these examples are for illustration only and do not limit the scope of the present invention. Variations of the present invention now known or further developed are considered to fall within the scope of the present invention described and claimed herein.

The compounds of the present invention are prepared using convenient starting materials and common preparation steps. The present invention provides typical or preferred reaction conditions, such as reaction temperature, time, solvent, pressure, and molar ratio of reactants. However, unless otherwise specified, other reaction conditions can also be adopted. The optimized conditions may change with the use of specific reactants or solvents, but in general, the reaction optimized steps and conditions can be determined.

In addition, some protecting groups may be used in the present invention to protect certain functional groups from unnecessary reactions. Protecting groups suitable for various functional groups and their protection or deprotection conditions are widely known to those skilled in the art. For example, T.W.Greene and G.M.Wuts' Protecting Groups in Organic Preparations (3rd edition, Wiley, New York, 1999 and references therein) describe in detail the protection or deprotection of a large number of protecting groups.

The separation and purification of compounds and intermediates can be carried out by appropriate methods and steps according to specific needs, such as filtration, extraction, distillation, crystallization, column chromatography, preparative thin layer chromatography, preparative high performance liquid chromatography or a combination of the above methods. The specific use method can refer to the examples described in the present invention. Of course, other similar separation and purification means can also be adopted. Conventional methods (including physical constants and spectral data) can be used to characterize them.

The structures of the compounds were determined by nuclear magnetic resonance (NMR) and/or mass spectrometry (MS). NMR shifts are given in units of 10 ^-6 (ppm). NMR measurements were performed using a Brukerdps 300 NMR spectrometer, with deuterated dimethyl sulfoxide (DMSO-d ₆ ), deuterated chloroform (CDCl ₃ ), deuterated methanol (CD ₃ OD) as the solvent, and tetramethylsilane (TMS) as the internal standard.

MS was measured using LC (Waters 2695)/MS (Quattro Premier xE) mass spectrometer (manufacturer: Waters) (Photodiode Array Detector).

The preparative high performance liquid chromatography method used an LC6000 high performance liquid chromatograph (manufacturer: Chuangxin Tongheng), the chromatographic column was Daisogel C18 10 μm 100A (30 mm×250 mm), and the mobile phase was acetonitrile/water.

The thin layer chromatography silica gel plate used was Qingdao Ocean Chemical GF254 silica gel plate. The silica gel plate used in thin layer chromatography (TLC) had a specification of 0.20 mm to 0.25 mm, and the specification used in preparing the thin layer chromatography separation and purification product was 0.5 mm.

Column chromatography generally uses Qingdao marine silica gel 100-200 mesh, 200-300 mesh and 300-400 mesh silica gel as the carrier.

The known starting materials of the present invention can be synthesized by methods known in the art, or can be purchased from online shopping malls, Beijing Coupling, Sigma, Bailingwei, Yishiming, Shanghai Shuya, Shanghai Inokai, Anaiji Chemical, Shanghai Bid and other companies.

Unless otherwise specified in the examples, all reactions were carried out under a nitrogen atmosphere.

Argon atmosphere or nitrogen atmosphere means that the reaction bottle is connected to an argon or nitrogen balloon with a volume of about 1L.

Reaction solvent, organic solvent or inert solvent are each expressed as the solvent used that does not participate in the reaction under the described reaction conditions, including benzene, toluene, acetonitrile, tetrahydrofuran (THF), dimethylformamide (DMF), chloroform, dichloromethane, ether, methanol, nitrogen-methylpyrrolidone (NMP), pyridine, etc. Unless otherwise specified in the examples, the solution refers to an aqueous solution.

The chemical reactions described in the present invention are generally carried out under normal pressure. The reaction temperature is between -78°C and 200°C. The reaction time and conditions are, for example, between -78°C and 200°C at one atmosphere, and are completed within about 1 to 24 hours. If the reaction is left overnight, the reaction time is generally 16 hours. In the embodiments, unless otherwise specified, the reaction temperature is room temperature, which is 20°C to 30°C.

The reaction progress in the examples was monitored by thin layer chromatography (TLC), and the developing solvent systems used in the reaction were: A: dichloromethane and methanol system, B: petroleum ether and ethyl acetate system, C: acetone, and the volume ratio of the solvents was adjusted according to the polarity of the compounds.

The eluent system of column chromatography and the developing solvent system of thin layer chromatography used for purifying compounds include: A: dichloromethane and methanol system, B: petroleum ether and ethyl acetate system. The volume ratio of the solvent is adjusted according to the polarity of the compound, and a small amount of alkaline or acidic reagents such as triethylamine and trifluoroacetic acid can also be added for adjustment.

Unless otherwise defined, all professional and scientific terms used herein have the same meaning as those familiar to those skilled in the art. In addition, any method and material similar or equivalent to the described content can be applied to the method of the present invention.

The compounds of the invention are prepared according to the exemplary procedures provided herein and variations thereof known to those skilled in the art.

Example

Example 1: Preparation of (S)-1-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-2-(4-chlorophenyl)-2-((S)-2-azaspiro[4.4]nonan-3-yl)ethan-1-one (1)

Step 1: Preparation of tert-butyl 4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazine-1-carboxylate (1a)

At room temperature, 4-chloro-7H-pyrrolo[2,3-d]pyrimidine (5.0 g, 32.7 mmol) and DMF (100 mL) were added to a reaction bottle, DIEA (6.3 g, 49.0 mmol) was added dropwise, and piperazine-1-carboxylic acid tert-butyl ester (6.7 g, 35.9 mmol) was added in batches. After the addition, the temperature was raised to 110°C and the reaction was allowed to proceed overnight. After cooling to room temperature, the reaction system was poured into an ice-water bath to precipitate a solid, which was filtered and dried to obtain 8.5 g of the title compound, with a yield of 85.8%.

LCMS: m/z 304.17 [M+H] ⁺ .

Step 2: Preparation of 4-(piperazin-1-yl)-7H-pyrrolo[2,3-d]pyrimidine dihydrochloride (1b)

At room temperature, tert-butyl 4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazine-1-carboxylate (1a) (100 mg, 0.331 mmol) and methanol (3 mL) were added to the reaction flask, cooled to about 5°C in an ice bath, and a solution of hydrogen chloride in 1,4-dioxane (1.65 mL, 6.6 mmol) was added dropwise. The reaction was continued at room temperature for 10 hours. After the reaction was completed, the mixture was concentrated directly to obtain 90 mg of a white solid crude title compound, which was directly used in the next step.

LCMS: m/z 204.12 [M+H] ⁺ .

Step 3: Preparation of methyl 2-cyclopentylidene acetate (1c)

Dissolve methyl 2-(diethoxyphosphono)acetate (32.5 g, 0.130 mol) in anhydrous tetrahydrofuran (80 mL), slowly add sodium hydride (3.71 g, 0.130 mol) in batches under an ice bath, and react at room temperature for 1 hour. Then add cyclopentanone (10.0 g, 0.120 mol) and stir at room temperature for 4 hours. The reaction solution is concentrated under reduced pressure, the residue is extracted with ethyl acetate and water, the organic phase is dried and concentrated under reduced pressure, and the residue is separated and purified by silica gel column chromatography (mobile phase: EA/PE=0:1-1:10) to obtain 11.9 g of the oily title compound, yield: 70.8%.

LCMS: m/z 141 [M+H] ⁺ .

Step 4: Preparation of methyl 2-(1-(nitromethyl)cyclopentyl)acetate (1d)

At room temperature, methyl 2-cyclopentylidene acetate (1c) (11.9 g, 0.850 mol) was added to dimethyl sulfoxide (50 mL), and then cesium carbonate (5.59, 0.17 mol) and nitromethane (7.33 g, 1.19 mol) were added in sequence, and stirred at 80°C overnight. The reaction solution was quenched with glacial acetic acid (2.04 g, 0.340 mol), extracted with methyl tert-butyl ether, the organic phase was dried, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: EA/PE = 0:1-1:10) to obtain 16.5 g of the title compound, yield: 96.6%.

LCMS: m/z 202 [M+H] ⁺ .

Step 5: Preparation of 2-azaspiro[4.4]nonan-3-one (1e)

At room temperature, methyl 2-(1-(nitromethyl)cyclopentyl)acetate (1d) (1.00 g, 4.98 mmol) was added to ethyl acetate (5 mL), and then palladium carbon catalyst (200 mg) was added. The mixture was stirred overnight at room temperature under a hydrogen atmosphere. The reaction solution was filtered, the organic phase was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: MeOH/DCM = 0:1-1:50) to obtain 550 mg of the title compound, with a yield of 79.4%.

LCMS: m/z 140 [M+H] ⁺ .

Step 6: Preparation of tert-butyl 3-oxo-2-azaspiro[4.4]nonane-2-carboxylate (1f)

At room temperature, 2-azaspiro[4.4]nonane-3-one (1e) (8.13 g, 58.4 mmol) was added to dichloromethane (50 mL), followed by di-tert-butyl dicarbonate (31.9 g, 146 mmol), triethylamine (17.8 g, 175 mmol) and 4-dimethylaminopyridine (DMAP) (1.43 g, 11.7 mmol), and stirred at room temperature for 2 hours. The reaction solution was extracted with dichloromethane, the organic phase was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: MTBE/DCM = 0:1-1:10) to obtain 12.0 g of the title compound, yield: 85.9%.

LCMS: m/z 184 [M-55] ⁺ .

¹ H NMR (400MHz, DMSO-d6) δ3.55 (d, J = 44.0Hz, 2H), 2.43 (d, J = 44.0Hz, 2H), 1.68-1.60 (m, 8H), 1.53 (s, 9H ).

Step 7: Preparation of tert-butyl 3-hydroxy-2-azaspiro[4.4]nonane-2-carboxylate (1 g)

At room temperature, tert-butyl 3-oxo-2-azaspiro[4.4]nonane-2-carboxylate (1f) (1.50 g, 6.27 mmol) and THF (23.0 mL) were added to a reaction flask, cooled to -78°C, and DIBAL-H (10 mL, 10.0 mmol) was added dropwise. After the addition, the mixture was kept at -78°C for 30 minutes. After the reaction was completed, the reaction was quenched with saturated aqueous sodium acetate solution (5 mL), warmed to room temperature, and saturated aqueous ammonium chloride solution (5 mL) and ethyl acetate (10 mL) were added and stirred for 10 minutes. The mixture was filtered, and the aqueous phase was extracted with ethyl acetate. The organic phases were combined and concentrated under reduced pressure to obtain 1.38 g of the title product as a yellow oil, which was used directly in the next step with a yield of 91.4%.

LCMS: m/z 242.17 [M+H] ⁺ .

Step 8: Preparation of tert-butyl 3-methoxy-2-azaspiro[4.4]nonane-2-carboxylate (1h)

At room temperature, tert-butyl 3-hydroxy-2-azaspiro[4.4]nonane-2-carboxylate (1 g) (1.38 g, 5.72 mmol), methanol (20 mL) and p-toluenesulfonic acid monohydrate (52.0 mg, 0.270 mmol) were added to a reaction flask and reacted at room temperature for 2 hours. After the reaction, the reaction system was poured into a saturated sodium bicarbonate aqueous solution and extracted with ethyl acetate. The organic phases were combined, concentrated under reduced pressure, and diluted with dichloromethane to prepare a 29 wt% solution, which was directly used in the next step.

LCMS: m/z 256.18 [M+H] ⁺ .

Step 9: Preparation of (R)-4-benzyl-3-(2-(4-chlorophenyl)acetyl)oxazolidin-2-one (1i)

At room temperature, 2-(4-chlorophenyl)acetic acid (10.0 g, 58.6 mmol), (R)-4-benzyloxazolidin-2-one (10.4 g, 58.6 mmol), and toluene (150 mL) were added to the reaction flask, and triethylamine (23.7 g, 235 mmol) was added dropwise. The mixture was cooled to about 5°C in an ice bath, and pivaloyl chloride (PVIC) (9.10 g, 75 mmol) was added dropwise. The mixture was refluxed for 10 hours. After the reaction, the reaction system was poured into water, the aqueous phase was extracted with ethyl acetate, the organic phases were combined, dried over anhydrous Na ₂ SO ₄ , filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by column chromatography (PE:EA=5:1) to obtain 8.70 g of the target product as a yellow oil, which was directly used in the next step with a yield of 45%.

LCMS: m/z 330.08 [M+H] ⁺ .

Step 10: Preparation of (S)-3-((S)-2-((R)-4-benzyl-2-oxooxazolidin-3-yl)-1-(4-chlorophenyl)-2-oxoethyl)-2-azaspiro[4.4]nonane-2-carboxylic acid tert-butyl ester (1j)

At room temperature, (R)-4-benzyl-3-(2-(4-chlorophenyl)acetyl)oxazolidin-2-one (1i) (1.86 g, 5.64 mmol) and dichloromethane (30 mL) were added to a reaction flask, cooled to -78°C, titanium tetrachloride (1.18 g, 6.23 mmol) was added dropwise, and DIPEA (878 mg, 6.79 mmol) was added dropwise after keeping the temperature for 30 minutes. After keeping the temperature for 30 minutes, a dichloromethane solution of tert-butyl 3-methoxy-2-azaspiro[4.4]nonane-2-carboxylate (1h) was added dropwise. After keeping the temperature for 30 minutes, the temperature was naturally raised to room temperature and the reaction was carried out for 1.5 hours. After the reaction, the reaction system was poured into water, the aqueous phase was extracted with dichloromethane, the organic phases were combined, dried over _{anhydrous Na2SO4} , filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by column chromatography (PE:EA=5:1) to obtain 1.18 g of the target product as a yellow oil, which was directly used in the next step with a yield of 59.6%.

LCMS: m/z 553.24 [M+H] ⁺ .

Step 11: Preparation of (S)-2-((S)-2-(tert-butyloxycarbonyl)-2-azaspiro[4.4]non-3-yl)-2-(4-chlorophenyl)acetic acid (1k)

LiOH (38 mg, 0.904 mmol), water (2 mL) and THF (4 mL) were added to a reaction flask at room temperature, and the temperature was cooled to about 5°C in an ice bath. 30% hydrogen peroxide (82.1 mg, 0.724 mmol) was added dropwise, and (S)-tert-butyl 3-((S)-2-((R)-4-benzyl-2-oxooxazolidin-3-yl)-1-(4-chlorophenyl)-2-oxoethyl)-2-azaspiro[4.4]nonane-2-carboxylate (1j) (200 mg, 0.362 mmol) was added in batches, and the reaction was carried out in an ice bath for 30 hours. After the reaction was completed, the reaction was quenched with a 12.5 wt % aqueous sodium sulfite solution, the pH was adjusted to 2 with a 27.0 wt % potassium bisulfate solution, the aqueous phase was extracted with ethyl acetate, the organic phases were combined and concentrated, and the residue was separated and purified by column chromatography (PE:EA=10:1) to obtain 105 mg of the target product as a yellow oil, which was directly used in the next step with a yield of 73.6%.

LCMS: m/z 394.91 [M+H] ⁺ .

Step 12: Preparation of (S)-tert-butyl 3-((S)-2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-1-(4-chlorophenyl)-2-oxoethyl)-2-azaspiro[4.4]nonane-2-carboxylate (11).

At room temperature, 4-(piperazin-1-yl)-7H-pyrrolo[2,3-d]pyrimidine dihydrochloride (1b) (90 mg, crude product) and dichloromethane (5 mL) were added to a reaction flask, N,N-diisopropylethylamine (427 mg, 3.31 mmol) was added, and the mixture was stirred for 5 minutes. (S)-2-((S)-2-(tert-butyloxycarbonyl)-2-azaspiro[4.4]non-3-yl)-2-(4-chlorophenyl)acetic acid (1k) (112.9 mg, 0.39 mmol) was added, and the mixture was stirred for 1 minute. Benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU) (151 mg, 0.40 mmol) was added, and the mixture was stirred for 12 hours. After the reaction was completed, the reaction mixture was concentrated under reduced pressure and the residue was separated and purified by column chromatography (DCM:MeOH=10:1) to obtain 250 mg of the crude title compound as a white solid.

LCMS: m/z 579.28 [M+H] ⁺ .

Step 13: Preparation of (S)-1-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-2-(4-chlorophenyl)-2-((S)-2-azaspiro[4.4]nonan-3-yl)ethan-1-one (1).

At room temperature, (S)-3-((S)-2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-1-(4-chlorophenyl)-2-oxoethyl)-2-azaspiro[4.4]nonane-2-carboxylic acid tert-butyl ester (1l) (250 mg, crude product) and dichloromethane (3 mL) were added to a reaction flask, cooled to about 5°C in an ice bath, and a solution of hydrogen chloride in 1,4-dioxane (0.33 mL, 1.32 mmol) was added dropwise. The reaction was allowed to react at room temperature for 10 hours. After the reaction was completed, the mixture was directly concentrated, and the residue was separated by preparative liquid chromatography (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um, 100 A, mobile phase: acetonitrile/water, gradient: 30%-80%) to obtain 110 mg of the title compound as a white solid, with a three-step yield of 69.7%.

LCMS: m/z 479.20 [M+H] ⁺ .

¹ H NMR (400MHz, CDCl3) δ = 10.83 (s, 1H), 8.25 (s, 1H), 7.31 (d, J = 8.5, 2H), 7.24 (s, 1H), 7.11 (s, 1H), 7.07 (d,J＝3.6,1H),6.38(d,J＝3.6,1H),4.11-3.90(m,4H),3.89-3.78 (m,1H),3.75-3.65(m,1H),3.64-3.52(m,2H),3.41(dd,J＝12.9,6.2,1H),3.36-3.26(m,1H),2.99(dd, J=23.8,10.5,2H),1.72-1.45(m,8H),1.36(dd,J=14.4,9.6,2H).

Example 2: Preparation of (2S)-1-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-2-(4-chlorophenyl)-2-((8S)-2-oxa-7-azaspiro[4.4]nonan-8-yl)ethan-1-one (2)

Step 1: Preparation of methyl (E)-2-(dihydrofuran-3(2H)-ylidene)acetate (2a)

Add methyl 2-(diethoxyphosphonyl)acetate (13.4 g, 63.9 mmol) to THF (50 mL), add sodium hydride (2.55 g, 63.9 mmol) in batches at 0°C, warm to room temperature after addition, and stir for 1 hour. Add dihydrofuran-3(2H)-one (5.00 g, 58.1 mmol) and react at room temperature for 4 hours. Concentrate the reaction solution under reduced pressure, extract with ethyl acetate, wash the organic phase with water and saturated brine, dry over anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and separate and purify the residue by silica gel column chromatography (mobile phase: EA/DCM=1:50) to obtain 4.89 g of the title compound as a yellow oil, yield: 60%.

Step 2: Preparation of methyl 2-(3-(nitromethyl)tetrahydrofuran-3-yl)acetate (2b)

At room temperature, (E)-2-(dihydrofuran-3(2H)-ylidene)acetic acid methyl ester (2a) (2.00 g, 14.1 mmol) was added to DMSO (10 mL), followed by cesium carbonate (918 mg, 2.81 mmol) and nitromethane (1.20 g, 19.7 mmol). The mixture was stirred at 80 °C overnight. The reaction was quenched with glacial acetic acid (0.322 mL, 5.60 mmol), extracted with dimethyl tert-butyl ether, the organic phase was washed with water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: MeOH/DCM=1:20) to obtain 1.11 g of the title compound as a light yellow oil, yield: 38.9%.

Step 3: Preparation of 2-oxa-7-azaspiro[4.4]nonan-8-one (2c)

At room temperature, methyl 2-(3-(nitromethyl)tetrahydrofuran-3-yl)acetate (2b) (388 mg, 1.91 mmol) was added to methanol (5 mL), and then Pd/C (38.8 mg) was added. The mixture was stirred at room temperature overnight under a hydrogen atmosphere. The reaction solution was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: MeOH/DCM=1:20) to obtain 100 mg of the title compound as a red oil, with a yield of 37%.

Step 4: Preparation of tert-butyl 8-oxo-2-oxa-7-azaspiro[4.4]nonane-7-carboxylate (2d)

At room temperature, 2-oxa-7-azaspiro[4.4]nonane-8-one (2c) (100 mg, 0.708 mmol) was added to tetrahydrofuran (10 mL), followed by triethylamine (0.31 mL, 2.10 mmol), DMAP (8.60 mg, 0.071 mmol), and di-tert-butyl dicarbonate (387 mg, 1.77 mmol). The mixture was stirred at room temperature overnight under a nitrogen atmosphere. The reaction solution was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: MeOH/DCM=1:50) to obtain 80 mg of the title compound as a light yellow solid, with a yield of 46.7%.

LCMS: m/z 264 [M+Na] ⁺ .

¹ H NMR (400MHz, CDCl3) δ3.94-3.91 (m, 2H), 3.84 (d, J = 8.8Hz, 1H), 3.64-3.58 (m, 3H), 2.48 (s, 2H), 2.12-1.98 (m,2H),1.53(s,9H).

Step 5: Preparation of tert-butyl 8-hydroxy-2-oxa-7-azaspiro[4.4]nonane-7-carboxylate (2e)

At room temperature, 8-oxo-2-oxa-7-azaspiro[4.4]nonane-7-carboxylic acid tert-butyl ester (2d) (1.10 g, 4.56 mmol) and THF (20.0 mL) were added to a reaction flask, cooled to -78°C, and DIBAL-H (10 mL, 10.0 mmol) was added dropwise. After the addition was completed, the mixture was kept at -78°C for 30 minutes. After the reaction was completed, the reaction was quenched with saturated aqueous sodium acetate solution (5 mL), warmed to room temperature, and saturated aqueous ammonium chloride solution (5 mL) and ethyl acetate (10 mL) were added and stirred for 10 minutes, filtered, and the aqueous phase was extracted with ethyl acetate. The organic phases were combined and concentrated under reduced pressure to obtain 600 mg of the title product as a yellow oily crude product, which was used directly in the next step.

LCMS: m/z 244.15 [M+H] ⁺ .

Step 6: Preparation of tert-butyl 8-methoxy-2-oxa-7-azaspiro[4.4]nonane-7-carboxylate (2f)

At room temperature, 8-hydroxy-2-oxa-7-azaspiro[4.4]nonane-7-carboxylic acid tert-butyl ester (2e) (600 mg, 2.47 mmol) methanol (12 mL) and p-toluenesulfonic acid monohydrate (52.0 mg, 0.270 mmol) were added to a reaction flask and reacted at room temperature for 2 hours. After the reaction, the reaction system was poured into a saturated sodium bicarbonate aqueous solution and extracted with ethyl acetate. The organic phases were combined and concentrated under reduced pressure. The residue was diluted with dichloromethane to prepare a 29 wt% solution, which was directly used in the next step.

LCMS: m/z 258.16 [M+H] ⁺ .

Step 7: Preparation of (8S)-8-((S)-2-((R)-4-benzyl-2-oxooxazolidin-3-yl)-1-(4-chlorophenyl)-2-oxoethyl)-2-oxa-7-azaspiro[4.4]nonane-7-carboxylic acid tert-butyl ester (2 g)

At room temperature, (R)-4-benzyl-3-(2-(4-chlorophenyl)acetyl)oxazolidin-2-one (1i) (814 mg, 2.47 mmol) and dichloromethane (16 mL) were added to a reaction flask, cooled to -78°C, titanium tetrachloride (515 mg, 2.72 mmol) was added dropwise, and DIPEA (383 mg, 2.96 mmol) was added dropwise after keeping the temperature for 30 minutes. After keeping the temperature for 30 minutes, a dichloromethane solution of tert-butyl 8-methoxy-2-oxa-7-azaspiro[4.4]nonane-7-carboxylate (2f) was added dropwise. After keeping the temperature for 30 minutes, the temperature was naturally raised to room temperature and the reaction was carried out for 1.5 hours. After the reaction, the reaction system was poured into water, the aqueous phase was extracted with dichloromethane, the organic phases were combined, dried over _{anhydrous Na2SO4} , filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by column chromatography (PE:EA=5:1) to obtain 165 mg of the target product as a yellow oil, which was directly used in the next step. The three-step yield was 10.7%.

LCMS: m/z 555.22 [M+H] ⁺ .

Step 8: Preparation of (2S)-2-((8S)-7-(tert-butyloxycarbonyl)-2-oxa-7-azaspiro[4.4]nonan-8-yl)-2-(4-chlorophenyl)acetic acid (2h)

LiOH (38 mg, 0.904 mmol), water (2 mL) and THF (4 mL) were added to a reaction flask at room temperature, and the mixture was cooled to about 5°C in an ice bath. 30% hydrogen peroxide (82.1 mg, 0.724 mmol) was added dropwise, and (8S)-8-((S)-2-((S)-5-benzyl-2-oxazolidin-3-yl)-1-(4-chlorophenyl)-2-oxoethyl)-2-oxa-7-azaspiro[4.4]nonane-7-carboxylic acid tert-butyl ester (2 g) (200 mg, 0.361 mmol) was added in batches, and the mixture was reacted in an ice bath for 30 hours. After the reaction was completed, the reaction was quenched with a 12.5 wt % aqueous sodium sulfite solution, the pH was adjusted to 2 with a 27.0 wt % potassium bisulfate solution, the aqueous phase was extracted with ethyl acetate, the organic phases were combined and concentrated under reduced pressure, and the residue was separated and purified by column chromatography (PE: EA = 10: 1) to obtain 105 mg of the target product as a yellow oil, which was directly used in the next step with a yield of 73.6%.

LCMS: m/z 396.15 [M+H] ⁺ .

Step 9: Preparation of (8S)-8-((S)-2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-1-(4-chlorophenyl)-2-oxoethyl)-2-oxa-7-azaspiro[4.4]nonane-7-carboxylic acid tert-butyl ester (2i)

At room temperature, 4-(piperazin-1-yl)-7H-pyrrolo[2,3-d]pyrimidine dihydrochloride (1b) (100 mg, 0.362 mmol) and dichloromethane (2.5 mL) were added to a reaction flask, and N,N-diisopropylethylamine (DIPEA) (206 mg, 1.55 mmol) was added. The mixture was stirred for 5 minutes, and (2S)-2-((8S)-7-(tert-butyloxycarbonyl)-2-oxa-7-azaspiro[4.4]non-8-yl)-2-(4-chlorophenyl)acetic acid (2h) (100 mg, crude product) was added. The mixture was stirred for 1 minute, and HBTU (102 mg, 0.270 mmol) was added. The mixture was stirred for 12 hours. After the reaction was completed, the mixture was concentrated under reduced pressure, and the residue was separated and purified by column chromatography (DCM:MeOH=10:1) to obtain 140 mg of the crude title compound as a white solid.

LCMS: m/z 581.26 [M+H] ⁺ .

Step 10: Preparation of (2S)-1-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-2-(4-chlorophenyl)-2-((8S)-2-oxa-7-azaspiro[4.4]nonan-8-yl)ethan-1-one (2)

At room temperature, compound 2i (140 mg, crude product) and dichloromethane (2 mL) were added to a reaction flask, cooled to about 5°C in an ice bath, and a solution of hydrogen chloride in 1,4-dioxane (0.33 mL, 1.32 mmol) was added dropwise. The reaction was allowed to react at room temperature for 10 hours. After the reaction was completed, the mixture was directly concentrated, and the residue was separated and purified by preparative HPLC (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um, 100 A, mobile phase: acetonitrile/water, gradient: 30%-80%) to obtain 14 mg of the title compound as a white solid, with a three-step yield of 14.0%.

LCMS: m/z 481.20 [M+H] ⁺ .

¹ H NMR (400MHz, CDCl3) δ = 10.83 (s, 1H), 8.25 (s, 1H), 7.31 (d, J = 8.5, 2H), 7.24 (s, 1H), 7.11 (s, 1H), 7.07 (d,J＝3.6,1H),6.38(d,J＝3.6,1H),4.11-3.90(m,4H),3.89-3.78(m,1H),3.75-3.65(m,1H),3.64- 3.52(m,2H),3.41(dd,J＝12.9,6.2,1H),3.36-3.26(m,1H),1.72-1.45(m,8H),1.36(dd,J＝14.4,9.6,2H) .

Example 3: Preparation of (2S)-1-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-2-((1S,2S)-3-azabicyclo[3.1.0]hexan-2-yl)-2-(4-chlorophenyl)ethan-1-one (3)

Step 1: Preparation of tert-butyl 3-azabicyclo[3.1.0]hexane-3-carboxylate (3a)

At room temperature, 3-azabicyclo[3.1.0]hexane (6.00 g, 50.0 mmol), tetrahydrofuran (50 ml), and water (100 ml) were added to a reaction flask, and di-tert-butyl dicarbonate (12.0 g, 55.0 mmol) and sodium carbonate (13.3 g, 125 mmol) were added, and the mixture was stirred for 5 hours. After the reaction, ethyl acetate and water were added to the reaction flask for extraction, and the organic phase was washed with a saturated sodium chloride solution, and the organic phase was concentrated to dryness to obtain 10.2 g of the title compound as a yellow oil, which was used directly in the next step.

LCMS: m/z 184.13 [M+H] ⁺ .

Step 2: Preparation of tert-butyl 2-oxo-3-azabicyclo[3.1.0]hexane-3-carboxylate (3b)

At room temperature, tert-butyl 3-azabicyclo[3.1.0]hexane-3-carboxylate (3a) (10.2 g, 50.0 mmol) and ethyl acetate (250 ml) were added to a reaction flask, and ruthenium dioxide monohydrate (2.66 g, 20.0 mmol) and sodium periodate (53.5 g, 250 mmol) in an aqueous solution (200 ml) were slowly added, and the mixture was stirred for 12 hours. After the reaction was completed, the mixture was filtered, ethyl acetate and water were added to the filtrate for extraction, the organic phase was concentrated to dryness, and the residue was purified by column chromatography (eluent: dichloromethane: methanol = 80:1) to obtain 5.10 g of the title compound as a colorless oil, with a yield of 51.7%.

LCMS: m/z 198.11 [M+H] ⁺ .

Step 3: Preparation of tert-butyl 2-hydroxy-3-azabicyclo[3.1.0]hexane-3-carboxylate (3c)

At room temperature, tert-butyl 2-oxo-3-azabicyclo[3.1.0]hexane-3-carboxylate (3b) (5.79 g, 29.4 mmol) and tetrahydrofuran (60 mL) were added to a reaction flask, cooled to -78°C, and DIBAL-H (1.0 M, 49 mL, 49.0 mmol) was added dropwise under a nitrogen atmosphere. The mixture was kept at -78°C for 30 minutes. After the reaction was completed, the reaction was quenched with a saturated sodium acetate solution (30 mL), the temperature was naturally raised to 30°C, a saturated ammonium chloride solution (100 mL) was added, stirred for 10 minutes, ethyl acetate (150 mL) was added, filtered, the aqueous phase was extracted with ethyl acetate, the organic phases were combined, washed with a saturated sodium chloride solution, the organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 6.66 g of the title compound as a light yellow oil, which was used directly in the next step.

LCMS: m/z 200.12 [M+H] ⁺ .

Step 4: Preparation of tert-butyl 2-methoxy-3-azabicyclo[3.1.0]hexane-3-carboxylate (3d)

At room temperature, tert-butyl 2-hydroxy-3-azabicyclo[3.1.0]hexane-3-carboxylate (3c) (6.66 g, 32.3 mmol) and methanol (70 mL) were added to a reaction flask, and p-toluenesulfonic acid monohydrate (430 mg, 2.26 mmol) was added, and the mixture was stirred for 2 hours. After the reaction, the reaction solution was concentrated under reduced pressure, and saturated sodium bicarbonate solution and ethyl acetate were added for extraction. The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 5.63 g of the title compound as a yellow oil, which was diluted with dichloromethane (20 mL) and used directly in the next step.

LCMS: m/z 214.14 [M+H] ⁺ .

Step 5: Preparation of (1S,2S)-2-((S)-2-((R)-4-benzyl-2-oxooxazolidin-3-yl)-1-(4-chlorophenyl)-2-oxoethyl)-3-azabicyclo[3.1.0]hexane-3-carboxylic acid tert-butyl ester (3e)

At room temperature, compound 1i (8.87 g, 27.0 mmol) and dichloromethane (90 mL) were added to a reaction flask, cooled to -78°C, titanium tetrachloride (5.12 g, 27.0 mmol) was added dropwise, and DIPEA (4.74 g, 36.8 mmol) was added dropwise after keeping warm for 30 minutes. After keeping warm for 30 minutes, a dichloromethane solution (18 mL) of tert-butyl 2-methoxy-3-azabicyclo[3.1.0]hexane-3-carboxylate (3d) (5.21 g, 24.5 mmol) was added dropwise. After keeping warm for 30 minutes, the temperature was naturally raised to room temperature and the reaction was carried out for 1.5 hours. After the reaction, saturated sodium bicarbonate solution was added, extracted with dichloromethane, the organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (eluent: dichloromethane) to obtain 4.61 g of the title compound as a white solid with a yield of 36.9%.

LCMS: m/z 511.19 [M+H] ⁺ .

Step 6: Preparation of (2S)-2-((2S)-3-(tert-butyloxycarbonyl)-3-azabicyclo[3.1.0]hexan-2-yl)-2-(4-chlorophenyl)acetic acid (3f)

Lithium hydroxide monohydrate (104 mg, 2.45 mmol), tetrahydrofuran (3 mL) and water (3.5 mL) were added to a reaction flask at room temperature, 30% hydrogen peroxide (66.7 mg, 1.96 mmol) was added, the temperature was lowered to 0°C, and a solution of (1S,2S)-tert-butyl 2-((S)-2-((R)-4-benzyl-2-oxooxazolidin-3-yl)-1-(4-chlorophenyl)-2-oxoethyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (3e) (500 mg, 0.980 mmol) in tetrahydrofuran (4 mL) was added, and the reaction was stirred at 0°C for 3 hours. To the reaction solution was added 12.5 wt % sodium sulfite solution (1.2 mL) at 10 ° C., stirred for 0.5 hour, and 27.0 wt % potassium hydrogen sulfate solution was added to adjust the pH to 2-3. Ethyl acetate was added for extraction, and the organic phase was concentrated to dryness. The residue was separated and purified by preparative high performance liquid chromatography (chromatographic column model: Daisogei 30 mm * 250 mm, C18, 10 um, 100A, mobile phase: acetonitrile / water, gradient: 30% -80%) to give 215 mg of the title compound as a white solid, with a yield of 62.6%.

LCMS: m/z 352.12 [M+H] ⁺ .

Step 7: Preparation of (1S,2S)-tert-butyl 2-(2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-1-(4-chlorophenyl)-2-oxoethyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (3 g).

At room temperature, compound 1b (367 mg, 0.351 mmol) and dichloromethane (3 mL) were added to a reaction flask, DIEA (394 mg, 3.05 mmol) was added, and the mixture was stirred for 10 minutes. (2S)-2-((1S, 2S)-3-(tert-butoxycarbonyl)-3-azabicyclo[3.1.0]hexane-2-yl)-2-(4-chlorophenyl)acetic acid (3f) (107 mg, 0.305 mmol) was added, and the mixture was stirred for 5 minutes. HBTU (139 mg, 0.366 mmol) was added, and the mixture was stirred for 12 hours. After the reaction was completed, the reaction solution was concentrated to dryness, and the residue was purified by column chromatography (eluent: dichloromethane: methanol = 20:1) to obtain 414 mg of the title compound as a brown oil, which was used directly in the next step.

LCMS: m/z 537.23 [M+H] ⁺ .

Step 8: Preparation of (2S)-1-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-2-((1S,2S)-3-azabicyclo[3.1.0]hexan-2-yl)-2-(4-chlorophenyl)ethan-1-one (3).

At room temperature, compound 3g (373mg, 0.285mmol) and dichloromethane (5mL) were added to a reaction flask, and a solution of hydrogen chloride in 1,4-dioxane (4.0M, 0.33ml, 1.29mmol) was added, and the mixture was stirred at 25°C for 12 hours. After the reaction, the reaction solution was concentrated to dryness, diluted with dichloromethane, and the pH was adjusted to 7-8 with a saturated sodium bicarbonate aqueous solution. The system was concentrated under reduced pressure, and the residue was separated by preparative liquid chromatography (chromatographic column model: Daisogei 30mm*250mm, C18, 10um, 100A, mobile phase: acetonitrile/water, gradient: 30%-80%) to obtain 77.0mg of the title compound as a white solid with a yield of 62.1%.

LCMS: m/z 437.18 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO) δ11.72 (s, 1H), 8.14 (s, 1H), 7.44 (m, 4H), 7.19 (dd, J = 3.4, 2.5Hz, 1H), 6.59 (dd, J ＝3.5,1.8Hz,1H),3.82(m,5H),3.66(t,J＝10.3Hz,4H),3.56(m,1H),2.89(dd,J＝11.0,3.0Hz,1H),2.72 (d,J＝11.1Hz,1H),1.28(m,1H),0.77(ddd,J＝7.9,6.2,3.9Hz,1H),0.23(ddt,J＝34.7,8.1,4.2Hz,2H).

Example 4: Preparation of (S)-1-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-2-(4-chlorophenyl)-2-((S)-4-azaspiro[2.4]heptane-5-yl)ethan-1-one (4)

Step 1: Preparation of 4-azaspiro[2.4]heptan-5-one (4a)

At room temperature, methyl 3-cyanopropionate (7.91 g, 70.0 mmol) and tetrahydrofuran (200 mL) were added to the reaction flask, and tetraisopropyl titanate (3.98 g, 14.0 mmol) was added. The temperature was lowered to below 10°C, and ethylmagnesium bromide (3.0 M, 51.4 mL, 154 mmol) was slowly added under nitrogen atmosphere. After the addition, the temperature was slowly raised to room temperature and stirred for 12 hours. After the reaction was completed, water (3.5 mL) was added to the reaction flask, stirred for 20 minutes, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (eluent: dichloromethane: methanol = 20:1) to obtain 10.3 g of the title compound as a brown oil, which was used directly in the next step.

LCMS: m/z 112.07 [M+H] ⁺ .

Step 2: Preparation of tert-butyl 5-oxo-4-azaspiro[2.4]heptane-4-carboxylate (4b)

At room temperature, 4-azaspiro[2.4]heptane-5-one (4a) (8.83 g, 60.0 mmol), di-tert-butyl dicarbonate (19.7 g, 90.0 mmol), and tetrahydrofuran (90 mL) were added to a reaction flask, cooled to below 5°C, and triethylamine (18.2 g, 180 mmol) and DMAP (366 mg, 3.00 mmol) were added, and stirred for 12 hours. After the reaction, ethyl acetate and water were added to the reaction flask for extraction, and the organic phase was concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 5:1) to obtain 4.92 g of the title compound as a light yellow solid, with a yield of 33.5%.

LCMS: m/z 212.12 [M+H] ⁺ .

Step 3: Preparation of tert-butyl 5-hydroxy-4-azaspiro[2.4]heptane-4-carboxylate (4c)

At room temperature, tert-butyl 5-oxo-4-azaspiro[2.4]heptane-4-carboxylate (4b) (7.81 g, 37.0 mmol) and tetrahydrofuran (90 mL) were added to the reaction flask, cooled to -78°C, and DIBAL-H (1.0 M, 44.4 mL, 44.4 mmol) was added dropwise under nitrogen atmosphere. After the addition, the mixture was kept at -78°C for 30 minutes. After the reaction was completed, the reaction was quenched with saturated sodium acetate solution (20 mL), naturally heated to 30°C, saturated ammonium chloride solution (70 mL) was added, stirred for 10 minutes, ethyl acetate (120 mL) was added, filtered, the aqueous phase was extracted with ethyl acetate, the organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 8.12 g of the title compound as a yellow oil, which was used directly in the next step.

LCMS: m/z 214.14 [M+H] ⁺ .

Step 4: Preparation of tert-butyl 5-methoxy-4-azaspiro[2.4]heptane-4-carboxylate (4d)

At room temperature, tert-butyl 5-hydroxy-4-azaspiro[2.4]heptane-4-carboxylate (4c) (8.12 g, 37.0 mmol) and methanol (90 mL) were added to a reaction flask, and p-toluenesulfonic acid monohydrate (493 mg, 2.59 mmol) was added, and the mixture was stirred for 2 hours. After the reaction, the reaction solution was concentrated under reduced pressure, and saturated sodium bicarbonate solution and ethyl acetate were added for extraction. The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 7.96 g of the title compound as a brown oil, which was diluted with dichloromethane (20 mL) and used directly in the next step.

LCMS: m/z 228.15 [M+H] ⁺ .

Step 5: Preparation of (S)-tert-butyl 5-((S)-2-((R)-4-benzyl-2-oxooxazolidin-3-yl)-1-(4-chlorophenyl)-2-oxoethyl)-4-azaspiro[2.4]heptane-4-carboxylate (4e)

At room temperature, compound 1i (9.71 g, 29.5 mmol) and dichloromethane (110 mL) were added to a reaction flask, cooled to -78°C, titanium tetrachloride (5.61 g, 29.5 mmol) was added dropwise, and DIPEA (5.71 g, 44.3 mmol) was added dropwise after keeping warm for 30 minutes. After keeping warm for 30 minutes, a dichloromethane solution (18 mL) of tert-butyl 5-methoxy-4-azaspiro[2.4]heptane-4-carboxylate (4d) (8.70 g, 38.4 mmol) was added dropwise. After keeping warm for 30 minutes, the temperature was naturally raised to room temperature and the reaction was carried out for 1.5 hours. After the reaction, saturated sodium bicarbonate solution and dichloromethane were added for extraction. The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 10:1) to obtain 12.3 g of the title compound as a white solid with a yield of 79.8%.

LCMS: m/z 525.21 [M+H] ⁺ .

Step 6: Preparation of (S)-2-((S)-4-(tert-butoxycarbonyl)-4-azaspiro[2.4]heptane-5-yl)-2-(4-chlorophenyl)acetic acid (4f)

Lithium hydroxide monohydrate (106 mg, 2.50 mmol), tetrahydrofuran (5 mL) and water (3.5 mL) were added to a reaction flask at room temperature, 30% hydrogen peroxide (68.0 mg, 2.00 mmol) was added, the temperature was lowered to below 0°C, (S)-5-((S)-2-((R)-4-benzyl-2-oxooxazolidin-3-yl)-1-(4-chlorophenyl)-2-oxoethyl)-4-azaspiro[2.4]heptane-4-carboxylic acid tert-butyl ester (4e) (524 mg, 1.00 mmol) in tetrahydrofuran (2 mL) was added, and the reaction was stirred at about 0°C for 3 hours. After the reaction, 12.5 wt% sodium sulfite solution (1.2 mL) was added to the reaction solution at below 10°C, stirred for 0.5 hours, 27.0 wt% potassium hydrogen sulfate solution was added to adjust the pH to 2-3, ethyl acetate was added for extraction, the organic phase was concentrated under reduced pressure, and the residue was separated by preparative high performance liquid chromatography (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um, 100A, mobile phase: acetonitrile/water, gradient: 30%-80%) to obtain 233 mg of the title compound as a white solid, with a yield of 63.8%.

LCMS: m/z 366.14 [M+H] ⁺ .

Step 7: Preparation of (5S)-tert-butyl 5-(2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-1-(4-chlorophenyl)-2-oxoethyl)-4-azaspiro[2.4]heptane-4-carboxylate (4 g).

At room temperature, compound 1b (481 mg, 0.502 mmol) and dichloromethane (5 mL) were added to a reaction flask, DIEA (587 mg, 4.55 mmol) was added, and the mixture was stirred for 10 minutes. (S)-2-((S)-4-(tert-butoxycarbonyl)-4-azaspiro[2.4]heptane-5-yl)-2-(4-chlorophenyl)acetic acid (4f) (166 mg, 0.455 mmol) was added, and the mixture was stirred for 5 minutes. HBTU (207 mg, 0.546 mmol) was added, and the mixture was stirred for 12 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane: methanol = 20:1) to obtain 266 mg of the title compound as a brown solid, which was used directly in the next step.

LCMS: m/z 551.25 [M+H] ⁺ .

Step 8: Preparation of (S)-1-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-2-(4-chlorophenyl)-2-((S)-4-azaspiro[2.4]heptan-5-yl)ethan-1-one (4).

At room temperature, compound 4g (239mg, 0.409mmol) and dichloromethane (5mL) were added to a reaction flask, and a solution of hydrogen chloride in 1,4-dioxane (4.0M, 0.46ml, 1.84mmol) was added, and the mixture was stirred at 25°C for 12 hours. After the reaction, the reaction solution was concentrated under reduced pressure, diluted with dichloromethane, and the pH was adjusted to 7-8 with DIEA. The system was concentrated under reduced pressure, and the residue was separated by preparative liquid chromatography (chromatographic column model: Daisogei 30mm*250mm, C18, 10um, 100A, mobile phase: acetonitrile/water, gradient: 30%-80%) to obtain 24.0mg of the title compound as a white solid, with a yield of 13.1%.

LCMS: m/z 451.19 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO) δ11.72(s,1H),8.14(s,1H),7.40(dd,J＝21.4,8.6Hz,4H),7.19(m,1H),6.58(dd,J ＝3.5,1.7Hz,1H),4.53(m,1H),4.00(d,J＝9.3Hz,1H),3.78(m,5H),3.60(m,3H),2.25(m,4H),1.63 (m, 1H), 1.29 (ddd, J = 19.8, 14.4, 10.3Hz, 1H), 1.01 (t, J = 7.5Hz, 3H).

Example 5: Preparation of (S)-1-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-2-(4-chlorophenyl)-2-((R)-2-azaspiro[4.5]dec-1-yl)ethan-1-one (5)

Step 1: Preparation of tert-butyl 1-oxo-2-azaspiro[4.5]decane-2-carboxylate (5a)

At room temperature, 2-azaspiro[4.5]decane-1-one (200 mg, 1.31 mmol), DCM (10 mL), TEA (331 mg, 3.3 mmol), and DMAP (16 mg, 0.13 mmol) were added to the reaction flask, cooled to 0°C, and di-tert-butyl dicarbonate (343 mg, 1.57 mmol) was added dropwise. The mixture was stirred for 30 hours. After the reaction, the mixture was poured into water, and the aqueous phase was extracted with dichloromethane. The organic phases were combined and concentrated under reduced pressure to obtain 338 mg of the title product as a crude yellow oil, which was used directly in the next step.

LCMS: m/z 254.17 [M+H] ⁺ .

Step 2: Preparation of tert-butyl 1-hydroxy-2-azaspiro[4.5]decane-2-carboxylate (5b)

At room temperature, tert-butyl 1-oxo-2-azaspiro[4.5]decane-2-carboxylate (5a) (338 mg, crude product) and THF (23.0 mL) were added to a reaction flask, cooled to -78°C, and DIBAL-H (2 mL, 2.0 mmol) was added dropwise. After the addition, the mixture was kept at -78°C for 30 minutes. After the reaction was completed, the reaction was quenched with a saturated aqueous sodium acetate solution (5 mL), warmed to room temperature, and a saturated aqueous ammonium chloride solution (5 mL) and ethyl acetate (10 mL) were added and stirred for 10 minutes. The mixture was filtered, and the aqueous phase was extracted with ethyl acetate. The organic phases were combined and concentrated under reduced pressure to obtain 270 mg of the title product as a yellow oil, which was used directly in the next step with a yield of 81%.

LCMS: m/z 256.18 [M+H] ⁺ .

Step 3: Preparation of tert-butyl 1-methoxy-2-azaspiro[4.5]decane-2-carboxylate (5c)

At room temperature, tert-butyl 1-hydroxy-2-azaspiro[4.5]decane-2-carboxylate (5b) (270 mg, 1.06 mmol), methanol (20 mL) and p-toluenesulfonic acid monohydrate (52.0 mg, 0.270 mmol) were added to a reaction flask and reacted at room temperature for 2 hours. After the reaction, the reaction system was poured into a saturated sodium bicarbonate aqueous solution and extracted with ethyl acetate. The organic phases were combined and concentrated under reduced pressure. The residue was diluted with dichloromethane to prepare a 29 wt% solution, which was directly used in the next step.

LCMS: m/z 270.39 [M+H] ⁺ .

Step 4: Preparation of (R)-1-((S)-2-((R)-4-benzyl-2-oxooxazolidin-3-yl)-1-(4-chlorophenyl)-2-oxoethyl)-2-azaspiro[4.5]decane-2-carboxylic acid tert-butyl ester (5d)

At room temperature, compound 1i (1.86 g, 5.64 mmol) and dichloromethane (30 mL) were added to a reaction flask, cooled to -78°C, titanium tetrachloride (1.18 g, 6.23 mmol) was added dropwise, and the mixture was kept warm for 30 minutes. DIPEA (878 mg, 6.79 mmol) was added dropwise, and a dichloromethane solution of 1-methoxy-2-azaspiro[4.5]decane-2-carboxylic acid tert-butyl ester (5c) was added dropwise after keeping warm for 30 minutes. The mixture was naturally warmed to room temperature and reacted for 1.5 hours. After the reaction was completed, the reaction system was poured into water, the aqueous phase was extracted with dichloromethane, the organic phases were combined, dried over anhydrous Na ₂ SO ₄ , filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by column chromatography (PE:EA=5:1) to obtain 294 mg of the target product as a yellow oil, which was directly used in the next step with a two-step yield of 49%.

LCMS: m/z 567.25 [M+H] ⁺ .

Step 5: Preparation of (S)-2-((R)-2-(tert-butyloxycarbonyl)-2-azaspiro[4.5]dec-1-yl)-2-(4-chlorophenyl)acetic acid (5e)

LiOH (48.9 mg, 1.164 mmol), water (2.5 mL) and THF (5 mL) were added to a reaction flask at room temperature, and the temperature was cooled to about 5°C in an ice bath. 30% hydrogen peroxide (105.6 mg, 0.932 mmol) was added dropwise, and (R)-tert-butyl 1-((S)-2-((S)-5-benzyl-2-oxooxazolidin-3-yl)-1-(4-chlorophenyl)-2-oxoethyl)-2-azaspiro[4.5]decane-2-carboxylate (5d) (264 mg, 0.466 mmol) was added in batches, and the reaction was carried out in an ice bath for 3 hours. After the reaction was completed, the reaction was quenched with a 12.5 wt % aqueous sodium sulfite solution, the pH was adjusted to 2 with a 27.0 wt % potassium bisulfate solution, the aqueous phase was extracted with ethyl acetate, the organic phases were combined, and the residue was concentrated under reduced pressure and purified by column chromatography (PE:EA=10:1) to obtain 84 mg of the target product as a yellow oil, which was directly used in the next step with a yield of 44.2%.

LCMS: m/z 408.19 [M+H] ⁺ .

Step 6: Preparation of (R)-tert-butyl 1-((S)-2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-1-(4-chlorophenyl)-2-oxoethyl)-2-azaspiro[4.5]decane-2-carboxylate (5f).

At room temperature, compound 1b (90 mg, crude product) and dichloromethane (5 mL) were added to a reaction flask, N,N-diisopropylethylamine (427 mg, 3.31 mmol) was added, and the mixture was stirred for 5 minutes. (S)-2-((R)-2-(tert-butyloxycarbonyl)-2-azaspiro[4.5]dec-1-yl)-2-(4-chlorophenyl)acetic acid (5e) (112.9 mg, 0.28 mmol) was added, and the mixture was stirred for 1 minute. Benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate (151 mg, 0.40 mmol) was added, and the mixture was stirred for 12 hours. After the reaction was completed, the mixture was concentrated under reduced pressure, and the residue was subjected to column chromatography to obtain 250 mg of the crude title compound as a white solid.

LCMS: m/z 593.29 [M+H] ⁺ .

Step 7: Preparation of (S)-1-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-2-(4-chlorophenyl)-2-((R)-2-azaspiro[4.5]dec-1-yl)ethan-1-one (5)

At room temperature, compound 5f (250 mg, crude product) and dichloromethane (3 mL) were added to a reaction flask, cooled to about 5°C in an ice bath, and a solution of hydrogen chloride in 1,4-dioxane (0.33 mL, 1.32 mmol) was added dropwise. The reaction was allowed to react at room temperature for 10 hours. After the reaction was completed, the mixture was directly concentrated, and the residue was separated by preparative liquid chromatography (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um, 100 A, mobile phase: acetonitrile/water, gradient: 30%-80%) to obtain 25 mg of the title compound as a white solid, with a three-step yield of 15.4%.

LCMS: m/z 493.20 [M+H] ⁺ .

¹ H NMR (400MHz, CDCl3) δ = 10.57 (s, 1H), 8.28 (s, 1H), 7.39 (d, J = 8.5, 2H), 7.32 (d, J = 8.5, 2H), 7.07 (d, J＝3.6,1H),6.41(d,J＝3.6,1H),4.23(d,J＝9.3,1H),4.14-4.03(m,1H),3.94(d,J＝6.3,1H),3.79 (d,J＝9.5,2H),3.73-3.65(m,1 H),3.60(d,J＝9.6,3H),3.49-3.37(m,1H),3.12-2.96(m,2H),2.16-2.01(m,1H),1.76(d,J＝11.0,1H ),1.67-1.50(m,3H),1.49-1.30(m,3H),1.31-1.09(m,2H),0.96(d,J＝12.3,1H),0.90-0.74(m,1H),0.35 (td,J=13.1,3.9,1H).

Example 6: Preparation of (S)-1-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-2-(4-chlorophenyl)-2-((R)-2-azaspiro[4.4]nonan-1-yl)ethan-1-one (6)

Step 1: Preparation of methyl 1-(cyanomethyl)cyclopentane-1-carboxylate (6a)

At room temperature, methyl cyclopentanecarboxylate (1.02 g, 8.00 mmol) and THF (20 mL) were added to the reaction flask. The temperature was lowered to -78 °C, LDA (4.4 mL, 8.80 mmol) was added dropwise, and the mixture was stirred for 1 hour. At -78 °C, bromoacetonitrile (1.14 g, 9.60 mmol) was added dropwise, and the mixture was heated to room temperature and stirred for 10 hours. After the reaction was completed, the reaction solution was poured into a saturated aqueous ammonium chloride solution, the aqueous phase was extracted with ethyl acetate, the organic phases were combined, and the residue was concentrated under reduced pressure. The residue was separated and purified by column chromatography (PE: EA = 10: 1) to obtain 900 mg of the title product as a white solid, which was directly used in the next step with a yield of 67.7%.

LCMS: m/z 168.09 [M+H] ⁺ .

Step 2: Preparation of 2-azaspiro[4,4]nonan-1-one (6b)

At room temperature, 1-(cyanomethyl)cyclopentane-1-carboxylic acid methyl ester (6a) (850 mg, 5.09 mmol), THF (20.0 mL), water (10.0 mL), and cobalt chloride hexahydrate (607 mg, 2.55 mmol) were added to a reaction flask and stirred to dissolve. The temperature was lowered to 0°C, and NaBH ₄ (962 mg, 25.44 mmol) was added in batches. After the addition was completed, the mixture was heated to room temperature and stirred for 10 hours. After the reaction was completed, water (5 mL) and ethyl acetate (10 mL) were added and stirred for 10 minutes, filtered, and the aqueous phase was extracted with ethyl acetate. The organic phases were combined and concentrated under reduced pressure. The residue was separated and purified by column chromatography (PE:EA=2:1) to obtain 298 mg of the title product as a white solid, which was directly used in the next step with a yield of 42.1%.

LCMS: m/z 140.10 [M+H] ⁺ .

Step 3: Preparation of tert-butyl 1-oxo-2-azaspiro[4.4]nonane-2-carboxylate (6c)

At room temperature, 2-azaspiro[4,4]nonan-1-one (6b) (270 mg, 1.94 mmol), DCM (10 mL), TEA (588.1 mg, 5.82 mmol), and DMAP (11.8 mg, 0.097 mmol) were added to the reaction flask. The temperature was lowered to 0°C, and di-tert-butyl dicarbonate (634.7 mg, 2.91 mmol) was added dropwise, and stirred for 30 hours. After the reaction was completed, the reaction solution was poured into water, the aqueous phase was extracted with dichloromethane, the organic phases were combined, and concentrated under reduced pressure. The residue was separated and purified by column chromatography (PE:EA=5:1) to obtain 460 mg of the title product as a white solid, which was directly used in the next step with a yield of 99%.

LCMS: m/z 240.15 [M+H] ⁺ .

Step 4: Preparation of tert-butyl 1-hydroxy-2-azaspiro[4.4]nonane-2-carboxylate (6d)

At room temperature, tert-butyl 1-oxo-2-azaspiro[4.4]nonane-2-carboxylate (6c) (460 mg, 1.92 mmol) and THF (10.0 mL) were added to the reaction flask. The temperature was lowered to -78 °C, and DIBAL-H (2.9 mL, 2.88 mmol) was added dropwise. After the addition, the mixture was stirred at -78 °C for 30 minutes. After the reaction was completed, the reaction was quenched with saturated aqueous sodium acetate solution (5 mL), warmed to room temperature, and saturated aqueous ammonium chloride solution (5 mL) and ethyl acetate (10 mL) were added and stirred for 10 minutes. The mixture was filtered, and the aqueous phase was extracted with ethyl acetate. The organic phases were combined and concentrated under reduced pressure. The residue was separated and purified by column chromatography (PE:EA=5:1) to obtain 460 mg of the title product as a yellow oil, which was used directly in the next step with a yield of 99%.

LCMS: m/z 242.17 [M+H] ⁺ .

Step 5: Preparation of tert-butyl 1-methoxy-2-azaspiro[4.4]nonane-2-carboxylate (6e)

At room temperature, tert-butyl 1-hydroxy-2-azaspiro[4.4]nonane-2-carboxylate (6d) (412 mg, 1.71 mmol), methanol (20 mL) and p-toluenesulfonic acid monohydrate (16.2 mg, 0.09 mmol) were added to a reaction flask and reacted at room temperature for 2 hours. After the reaction, the reaction system was poured into a saturated sodium bicarbonate aqueous solution and extracted with ethyl acetate. The organic phases were combined and concentrated under reduced pressure. The residue was diluted with dichloromethane to prepare a 29 wt% solution, which was directly used in the next step.

LCMS: m/z 256.18 [M+H] ⁺ .

Step 6: Preparation of (R)-1-((S)-2-((R)-4-benzyl-2-oxooxazolidin-3-yl)-1-(4-chlorophenyl)-2-oxoethyl)-2-azaspiro[4.4]nonane-2-carboxylic acid tert-butyl ester (6f)

At room temperature, compound 1i (627.9 mg, 1.91 mmol) and dichloromethane (30 mL) were added to a reaction flask, cooled to -78°C, titanium tetrachloride (398.4 mg, 2.10 mmol) was added dropwise, stirred for 30 minutes, and DIPEA (295.4 mg, 2.29 mmol) was added dropwise. After keeping the temperature for 30 minutes, a dichloromethane solution of 1-methoxy-2-azaspiro[4.4]nonane-2-carboxylic acid tert-butyl ester (6e) was added dropwise. After keeping the temperature for 30 minutes, the temperature was naturally raised to room temperature and the reaction was carried out for 1.5 hours. After the reaction was completed, the reaction system was poured into water, the aqueous phase was extracted with dichloromethane, the organic phases were combined, dried over anhydrous Na ₂ SO ₄ , filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by column chromatography (PE:EA=5:1) to obtain 543 mg of the target product as a yellow oil, which was directly used in the next step with a two-step yield of 59.9%.

LCMS: m/z 553.24 [M+H] ⁺ .

Step 7: Preparation of (S)-2-((R)-2-(tert-butoxycarbonyl)-2-azaspiro[4.4]non-1-yl)-2-(4-chlorophenyl)acetic acid (6 g)

At room temperature, LiOH (103 mg, 2.45 mmol), water (5 mL), and THF (10 mL) were added to the reaction flask. The mixture was cooled to about 5°C in an ice bath, 30% hydrogen peroxide (222 mg, 1.96 mmol) was added dropwise, and (R)-1-((S)-2-((S)-5-benzyl-2-oxooxazolidin-3-yl)-1-(4-chlorophenyl)-2-oxoethyl)-2-azaspiro[4.4]nonane-2-carboxylic acid tert-butyl ester (6f) (543 mg, 0.982 mmol) was added in batches, and the mixture was reacted in an ice bath for 3 hours. After the reaction, the reaction was quenched with a 12.5 wt % aqueous sodium sulfite solution, the pH was adjusted to 2 with a 27.0 wt % potassium bisulfate solution, the aqueous phase was extracted with ethyl acetate, the organic phases were combined and concentrated under reduced pressure, and the residue was separated and purified by column chromatography (PE:EA=10:1) to obtain 102 mg of the target product as a yellow oil, which was directly used in the next step with a yield of 31%.

LCMS: m/z 394.17 [M+H] ⁺ .

Step 8: Preparation of (R)-1-((S)-2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-1-(4-chlorophenyl)-2-oxoethyl)-2-azaspiro[4.4]nonane-2-carboxylic acid tert-butyl ester (6h)

At room temperature, compound 1b (68.7 mg, 0.293 mmol) and dichloromethane (5 mL) were added to a reaction flask, N,N-diisopropylethylamine (327.7 mg, 2.54 mmol) was added, and the mixture was stirred for 5 minutes. (S)-2-((R)-2-(tert-butyloxycarbonyl)-2-azaspiro[4.4]non-1-yl)-2-(4-chlorophenyl)acetic acid (6 g) (100 mg, 0.254 mmol) was added, and the mixture was stirred for 1 minute. HBTU (115.6 mg, 0.305 mmol) was added, and the mixture was stirred for 12 hours. After the reaction was completed, the mixture was concentrated under reduced pressure, and the residue was separated and purified by column chromatography (DCM:MeOH=10:1) to obtain 163 mg of the crude title compound as a white solid.

LCMS: m/z 579.28 [M+H] ⁺ .

Step 9: Preparation of (S)-1-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-2-(4-chlorophenyl)-2-((R)-2-azaspiro[4.4]nonan-1-yl)ethan-1-one (6)

At room temperature, compound 6h (163 mg, crude product) and dichloromethane (3 mL) were added to the reaction flask, cooled to about 5°C in an ice bath, and hydrochloric acid/dioxane solution (0.67 mL, 2.68 mmol) was added dropwise. The reaction was allowed to react at room temperature for 10 hours. After the reaction was completed, the mixture was directly concentrated, and the residue was separated by preparative HPLC (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um, 100 A, mobile phase: acetonitrile/water, gradient: 30%-80%) to obtain 51 mg of the title compound as a white solid, with a yield of 39.4%.

LCMS: m/z 479.26 [M+H] ⁺ .

¹ H NMR (400MHz, CDCl3) δ = 10.83 (s, 1H), 8.25 (s, 1H), 7.31 (d, J = 8.5, 2H), 7.24 (s, 1H), 7.11 (s, 1H), 7.07 (d,J＝3.6,1H),6.38(d,J＝3.6,1H),4.11-3.90(m,4H),3.89-3.78 (m,1H),3.75-3.65(m,1H),3.64-3.52(m,2H),3.41(dd,J＝12.9,6.2,1H),3.36-3.26(m,1H),2.99(dd, J=23.8,10.5,2H),1.72-1.45(m,8H),1.36(dd,J=14.4,9.6,2H).

Example 7: Preparation of (S)-1-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-2-(4-chlorophenyl)-2-((R)-5-azaspiro[2.4]heptane-4-yl)ethan-1-one (7)

Step 1: Preparation of methyl 1-(hydroxymethyl)cyclopropane-1-carboxylate (7a)

At room temperature, dimethyl cyclopropane-1,1-dicarboxylate (10.0 g, 63.3 mmol) and THF (150 mL) were added to a reaction flask, and lithium tri(tert-butoxy)aluminum hydride (35.4 g, 139.2 mmol) was added in batches. After the addition, the temperature was raised to 60°C and the reaction was allowed to react overnight. The mixture was cooled to room temperature, and the reaction was quenched with a 10% KHSO ₄ aqueous solution. The mixture was extracted with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: EA/PE=0:1-1:10) to obtain 5.8 g of the title compound, with a yield of 70.7%.

LCMS: m/z 131.06 [M+H] ⁺ .

Step 2: Preparation of methyl 1-((p-toluenesulfonyloxy)methyl)cyclopropane-1-carboxylate (7b)

At room temperature, 1-(hydroxymethyl)cyclopropane-1-carboxylic acid methyl ester (5.0 g, 38.4 mmol), DCM (100 mL), TEA (7.76 g, 76.8 mmol), DMAP (468 mg, 3.84 mmol) were added to the reaction bottle in sequence, and p-toluenesulfonyl chloride (8.8 g, 46.08 mmol) was added in batches, and the reaction was allowed to react overnight at room temperature. The reaction system was poured into ice water to quench, the aqueous phase was extracted with DCM, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: EA/PE = 0:1-1:10) to obtain 6.0 g of the title compound, yield: 55.0%.

LCMS: m/z 285.07 [M+H] ⁺ .

Step 3: Preparation of methyl 1-(cyanomethyl)cyclopropane-1-carboxylate (7c)

At room temperature, 1-((toluenesulfonyloxy)methyl)cyclopropane-1-carboxylic acid methyl ester (7b) (5.0 g, 17.6 mmol) and THF (100 mL) were added to a reaction flask, and 1M trimethylsilyl cyanide (TMSCN) (5.24 g, 52.8 mmol) and 1M tetrabutylammonium fluoride (TBAF) THF solution (52.8 mL, 52.8 mmol) were added dropwise, and the reaction was allowed to proceed overnight at room temperature. The reaction system was poured into ice water and ethyl acetate to quench the reaction, the aqueous phase was extracted with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: EA/PE = 0:1-1:10) to obtain 2.25 g of the title compound, yield: 92.2%.

LCMS: m/z 140.06 [M+H] ⁺ .

Step 4: Preparation of 5-azaspiro[2.4]heptan-4-one (7d)

At room temperature, 1-(cyanomethyl)cyclopropane-1-carboxylic acid methyl ester (7c) (2.2 g, 15.82 mmol), THF (44 mL), and water (22 mL) were added to a reaction flask, and cobalt chloride hexahydrate (1.88 g, 7.91 mmol) was added. The mixture was cooled to 0°C in an ice bath, and sodium borohydride (2.99 g, 79.09 mmol) was added in batches. The reaction was allowed to react overnight at room temperature. The reaction system was poured into ice water and ethyl acetate to quench the reaction. The aqueous phase was extracted with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: EA/PE = 0:1-1:1) to obtain the title compound as a white solid (1.34 g, yield: 76.2%).

LCMS: m/z 112.07 [M+H] ⁺ .

Step 5: Preparation of tert-butyl 4-oxo-5-azaspiro[2.4]heptane-5-carboxylate (7e)

At room temperature, 5-azaspiro[2.4]heptane-4-one (7d) (1.24 g, 11.2 mmol) was added to dichloromethane (25 mL), followed by di-tert-butyl dicarbonate (3.7 g, 16.8 mmol), TEA (3.4 g, 33.6 mmol) and 4-dimethylaminopyridine (68.3 mg, 0.56 mmol), and stirred at room temperature for 2 hours. The reaction solution was extracted with dichloromethane, the organic phase was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: MTBE/DCM = 0:1-1:10) to obtain 2.24 g of the title compound, yield: 95.3%.

LCMS: m/z 156.12 [M-55] ⁺ .

Step 6: Preparation of tert-butyl 4-hydroxy-5-azaspiro[2.4]heptane-5-carboxylate (7f)

At room temperature, tert-butyl 4-oxo-5-azaspiro[2.4]heptane-5-carboxylate (7e) (1.0 g, 4.74 mmol) and THF (23.0 mL) were added to the reaction flask. The temperature was lowered to -78°C, and DIBAL-H (10 mL, 10.0 mmol) was added dropwise. After the addition was completed, the mixture was stirred at -78°C for 30 minutes. After the reaction was completed, the reaction was quenched with saturated aqueous sodium acetate solution (5 mL), warmed to room temperature, and saturated aqueous ammonium chloride solution (5 mL) and ethyl acetate (10 mL) were added and stirred for 10 minutes. The mixture was filtered, and the aqueous phase was extracted with ethyl acetate. After the organic phases were combined, the mixture was concentrated under reduced pressure to obtain 865 mg of the title product as a yellow oil, which was used directly in the next step with a yield of 86.5%.

LCMS: m/z 214.14 [M+H] ⁺ .

Step 7: Preparation of tert-butyl 4-methoxy-5-azaspiro[2.4]heptane-5-carboxylate (7 g)

At room temperature, tert-butyl 4-hydroxy-5-azaspiro[2.4]heptane-5-carboxylate (7f) (822 mg, 3.86 mmol), methanol (20 mL) and p-toluenesulfonic acid monohydrate (36.6 mg, 0.19 mmol) were added to a reaction flask and reacted at room temperature for 2 hours. After the reaction, the reaction system was poured into a saturated sodium bicarbonate aqueous solution and extracted with ethyl acetate. The organic phases were combined and concentrated under reduced pressure. The residue was diluted with dichloromethane to prepare a 29 wt% solution, which was directly used in the next step.

LCMS: m/z 228.15 [M+H] ⁺ .

Step 8: Preparation of (R)-4-((S)-2-((R)-4-benzyl-2-oxooxazolidin-3-yl)-1-(4-chlorophenyl)-2-oxoethyl)-5-azaspiro[2.4]heptane-5-carboxylic acid tert-butyl ester (7h)

At room temperature, compound 1i (1.7 g, 5.28 mmol) and dichloromethane (30 mL) were added to the reaction flask. The temperature was lowered to -78 °C, titanium tetrachloride (1.1 g, 5.81 mmol) was added dropwise, and DIPEA (818 mg, 6.34 mmol) was added dropwise after 30 minutes of incubation. After 30 minutes of incubation, a dichloromethane solution of 4-methoxy-5-azaspiro[2.4]heptane-5-carboxylic acid tert-butyl ester (7 g) was added dropwise, and the temperature was naturally raised to room temperature for 1.5 hours of reaction. After the reaction was completed, the reaction system was poured into water, the aqueous phase was extracted with dichloromethane, the organic phases were combined, dried over anhydrous Na ₂ SO ₄ , filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by column chromatography (PE:EA=5:1) to obtain 1.3 g of the target product as a yellow solid, which was directly used in the next step with a yield of 48.1%.

LCMS: m/z 525.21 [M+H] ⁺ .

Step 9: Preparation of (S)-2-((R)-5-(tert-butyloxycarbonyl)-5-azaspiro[2.4]heptane-4-yl)-2-(4-chlorophenyl)acetic acid (7i)

At room temperature, LiOH (247 mg, 5.88 mmol), water (13 mL), and THF (25 mL) were added to the reaction flask. The mixture was cooled to about 5°C in an ice bath, 30% hydrogen peroxide (533 mg, 4.70 mmol) was added dropwise, and (R)-4-((S)-2-((S)-5-benzyl-2-oxooxazolidin-3-yl)-1-(4-chlorophenyl)-2-oxoethyl)-5-azaspiro[2.4]heptane-5-carboxylic acid tert-butyl ester (7h) (1.3 g, 2.35 mmol) was added in batches, and the mixture was reacted in an ice bath for 30 hours. After the reaction, the reaction was quenched with a 12.5 wt % aqueous sodium sulfite solution, the pH was adjusted to 2 with a 27.0 wt % potassium bisulfate solution, the aqueous phase was extracted with ethyl acetate, the organic phases were combined and concentrated under reduced pressure, and the residue was separated by column chromatography (PE:EA=10:1) to obtain 90 mg of the target product as a white solid, which was directly used in the next step with a yield of 9.9%.

LCMS: m/z 366.14 [M+H] ⁺ .

Step 10: Preparation of (R)-4-((S)-2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-1-(4-chlorophenyl)-2-oxoethyl)-5-azaspiro[2.4]heptane-5-carboxylic acid tert-butyl ester (7j)

At room temperature, compound 1b (66.7 mg, 0.285 mmol) and dichloromethane (5 mL) were added to a reaction flask, N,N-diisopropylethylamine (412 mg, 3.10 mmol) was added, and the mixture was stirred for 5 minutes. (S)-2-((R)-5-(tert-butyloxycarbonyl)-5-azaspiro[2.4]heptane-4-yl)-2-(4-chlorophenyl)acetic acid (7i) (90 mg, 0.247 mmol) was added, and the mixture was stirred for 1 minute. HBTU (112 mg, 0.296 mmol) was added, and the mixture was stirred for 12 hours. After the reaction was completed, the mixture was concentrated under reduced pressure, and the residue was separated and purified by column chromatography (DCM:MeOH=10:1) to obtain 256 mg of the crude title compound as a white solid.

LCMS: m/z 551.25 [M+H] ⁺ .

Step 11: Preparation of (S)-1-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-2-(4-chlorophenyl)-2-((R)-5-azaspiro[2.4]heptane-4-yl)ethan-1-one (7)

At room temperature, compound 7j (256 mg, crude product) and dichloromethane (3 mL) were added to a reaction flask, cooled to about 5°C in an ice bath, and hydrochloric acid/dioxane solution (0.33 mL, 1.32 mmol) was added dropwise. The reaction was allowed to react at room temperature for 10 hours. After the reaction was completed, the mixture was directly concentrated, and the residue was separated and purified by preparative HPLC (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um, 100 A, mobile phase: acetonitrile/water, gradient: 30%-80%) to obtain 84 mg of the title compound as a white solid, with a two-step yield of 70.8%.

LCMS: m/z 451.19 [M+H] ⁺ .

¹ H NMR (400MHz, CDCl3) δ = 10.63 (s, 1H), 8.27 (s, 1H), 7.35-7.28 (m, 2H), 7.24 (s, 2H), 7.08 (d, J = 3.6, 1H) ,6.41(d,J＝3.7,1H),4.13-4.03(m,1H),3.99(ddd,J＝16.7,11.5,6.4,3H),3.93-3.85(m,1H),3.76 -3.67(m,1H),3.60(ddd,J＝14.4,7.5,3.7,2H),3.50-3.41(m,1H),3.36-3.27(m,1H),3.01(q,J＝9.8,2H ),1.79-1.67(m,1H),1.29-1.18(m,1H),0.68-0.58(m,1H),0.58-0.51(m,1H),0.51-0.40(m,2H).

Example 8: Preparation of (S)-1-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-2-(4-chlorophenyl)-2-((R)-6-azaspiro[3.4]octan-5-yl)ethan-1-one (8)

Step 1: Preparation of dimethyl cyclobutane-1,1-dicarboxylate (8a)

At room temperature, cyclobutane-1,1-dicarboxylic acid (20.0 g, 138.85 mmol), methanol (150 mL), and concentrated hydrochloric acid (0.3 mL) were added to the reaction flask in sequence, and the temperature was raised to 60°C and stirred overnight. The mixture was cooled to room temperature, concentrated under reduced pressure, diluted with DCM, and the organic phase was washed twice with water, concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: EA/PE = 0:1-1:10) to obtain 21.7 g of the title compound, with a yield of 90.7%.

LCMS: m/z 173.07.06[M+H] ⁺ .

Step 2: Preparation of methyl 1-(hydroxymethyl)cyclobutane-1-carboxylate (8b)

At room temperature, dimethyl cyclobutane-1,1-dicarboxylate (8a) (15.0 g, 87.1 mmol) and THF (150 mL) were added to a reaction flask, and lithium tri(tert-butoxy)aluminum hydride (LTBA) (55.4 g, 217.9 mmol) was added in batches. After the addition, the temperature was raised to 60°C and stirred overnight. The mixture was cooled to room temperature, quenched with 10% KHSO ₄ aqueous solution, extracted with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: EA/PE=0:1-1:10) to obtain 8.6 g of the title compound, with a yield of 68.4%.

LCMS: m/z 145.08 [M+H] ⁺ .

Step 3: Preparation of methyl 1-((p-toluenesulfonyloxy)methyl)cyclobutane-1-carboxylate (8c)

At room temperature, 1-(Hydroxymethyl)cyclobutane-1-carboxylic acid methyl ester (8b) (7.5 g, 52.0 mmol), DCM (100 mL), TEA (7.76 g, 76.8 mmol), DMAP (468 mg, 3.84 mmol) were added to the reaction flask in sequence, and p-toluenesulfonyl chloride (9.9 g, 52.0 mmol) was added in batches, and stirred at room temperature overnight. The reaction system was poured into ice water to quench the reaction, the aqueous phase was extracted with DCM, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: EA/PE = 0:1-1:10) to obtain 11.7 g of the title compound, yield: 75.4%.

LCMS: m/z 299.09 [M+H] ⁺ .

Step 4: Preparation of methyl 1-(cyanomethyl)cyclobutane-1-carboxylate (8d)

At room temperature, 1-((p-toluenesulfonyloxy)methyl)cyclobutane-1-carboxylic acid methyl ester (8c) (11.0 g, 36.9 mmol) and THF (100 mL) were added to a reaction flask, TMSCN (10.9 g, 110.7 mmol) and 1M TBAF THF solution (110.7 mL, 110.7 mmol) were added dropwise, and stirred at room temperature overnight. The reaction system was poured into ice water and ethyl acetate to quench the reaction, the aqueous phase was extracted with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: EA/PE = 0:1-1:10) to obtain 9.6 g of the crude title compound, which was directly used in the next step.

LCMS: m/z 154.08 [M+H] ⁺ .

Step 5: Preparation of 6-azaspiro[3.4]octan-5-one (8e)

At room temperature, 1-(cyanomethyl)cyclobutane-1-carboxylic acid methyl ester (8d) (6.0 g, crude product), THF (80 mL), and water (40 mL) were added to a reaction flask, and cobalt chloride hexahydrate (4.7 g, 19.6 mmol) was added. The mixture was cooled to 0°C in an ice bath, and sodium borohydride (7.4 g, 195.9 mmol) was added in batches. After the addition was completed, the mixture was warmed to room temperature and stirred overnight. The reaction system was poured into ice water and ethyl acetate to quench the reaction. The aqueous phase was extracted with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: EA/PE = 0:1-1:1) to obtain 1.0 g of the title compound as a white solid, with a two-step yield of 34.7%.

LCMS: m/z 126.08 [M+H] ⁺ .

Step 6: Preparation of tert-butyl 5-oxo-6-azaspiro[3.4]octane-6-carboxylate (8f)

At room temperature, 6-azaspiro[3.4]octan-5-one (8e) (1.0 g, 8.0 mmol) was added to dichloromethane (25 mL), followed by di-tert-butyl dicarbonate (3.7 g, 16.8 mmol), triethylamine (3.4 g, 33.6 mmol) and 4-dimethylaminopyridine (68.3 mg, 0.56 mmol), and stirred at room temperature for 2 hours. The reaction solution was extracted with dichloromethane, the organic phase was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: MTBE/DCM = 0:1-1:10) to obtain 1.40 g of the title compound, yield: 77.7%.

LCMS: m/z 170.14 [M-55] ⁺ .

Step 7: Preparation of tert-butyl 5-hydroxy-6-azaspiro[3.4]octane-6-carboxylate (8 g)

At room temperature, tert-butyl 5-oxo-6-azaspiro[3.4]octane-6-carboxylate (8f) (1.0 g, 3.92 mmol) and THF (23.0 mL) were added to the reaction flask. The temperature was lowered to -78 °C, and DIBAL-H (10 mL, 10.0 mmol) was added dropwise. After the addition was completed, the mixture was stirred at -78 °C for 30 minutes. After the reaction was completed, the reaction was quenched with saturated aqueous sodium acetate solution (5 mL), warmed to room temperature, and saturated aqueous ammonium chloride solution (5 mL) and ethyl acetate (10 mL) were added and stirred for 10 minutes. The mixture was filtered, and the aqueous phase was extracted with ethyl acetate. After the organic phases were combined, the mixture was concentrated under reduced pressure to obtain 865 mg of the title product as a yellow oil, which was used directly in the next step with a yield of 86.4%.

LCMS: m/z 228.15 [M+H] ⁺ .

Step 8: Preparation of tert-butyl 5-methoxy-6-azaspiro[3.4]octane-6-carboxylate (8h)

At room temperature, tert-butyl 5-hydroxy-6-azaspiro[3.4]octane-6-carboxylate (8g) (822mg, 3.62mmol), methanol (20mL) and p-toluenesulfonic acid monohydrate (36.6mg, 0.19mmol) were added to the reaction bottle and reacted at room temperature for 2 hours. After the reaction, the reaction system was poured into a saturated sodium bicarbonate aqueous solution and extracted with ethyl acetate. The organic phases were combined and concentrated under reduced pressure. The residue was diluted with dichloromethane to a 29wt% solution and used directly in the next step.

LCMS: m/z 242.17 [M+H] ⁺ .

Step 9: Preparation of (R)-5-((S)-2-((R)-4-benzyl-2-oxooxazolidin-3-yl)-1-(4-chlorophenyl)-2-oxoethyl)-6-azaspiro[3.4]octane-6-carboxylic acid tert-butyl ester (8i)

At room temperature, compound 1i (1.7 g, 5.28 mmol) and dichloromethane (30 mL) were added to a reaction flask, cooled to -78°C, titanium tetrachloride (1.1 g, 5.81 mmol) was added dropwise, and DIPEA (818 mg, 6.34 mmol) was added dropwise after 30 minutes of incubation. After 30 minutes of incubation, a dichloromethane solution of 5-methoxy-6-azaspiro[3.4]octane-6-carboxylic acid tert-butyl ester (8h) was added dropwise, and the temperature was naturally raised to room temperature for 1.5 hours of reaction. After the reaction was completed, the reaction system was poured into water, the aqueous phase was extracted with dichloromethane, the organic phases were combined, dried over anhydrous Na ₂ SO ₄ , filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by column chromatography (PE:EA=5:1) to obtain 1.3 g of the target product as a yellow solid, which was directly used in the next step with a yield of 46.7%.

LCMS: m/z 539.22 [M+H] ⁺ .

Step 10: Preparation of (S)-2-((R)-6-(tert-butyloxycarbonyl)-6-azaspiro[3.4]octan-5-yl)-2-(4-chlorophenyl)acetic acid (8j)

At room temperature, LiOH (247 mg, 5.88 mmol), water (13 mL), and THF (25 mL) were added to the reaction flask. The mixture was cooled to about 5°C in an ice bath, 30% hydrogen peroxide (533 mg, 4.70 mmol) was added dropwise, and (R)-5-((S)-2-((S)-5-benzyl-2-oxooxazolidin-3-yl)-1-(4-chlorophenyl)-2-oxoethyl)-6-azaspiro[3.4]octane-6-carboxylic acid tert-butyl ester (8i) (1.3 g, 2.41 mmol) was added in batches, and the mixture was reacted in an ice bath for 30 hours. After the reaction, the reaction was quenched with a 12.5 wt % aqueous sodium sulfite solution, the pH was adjusted to 2 with a 27.0 wt % potassium bisulfate solution, the aqueous phase was extracted with ethyl acetate, the organic phases were combined and concentrated under reduced pressure, and the residue was separated and purified by column chromatography (PE: EA = 10: 1) to obtain 90 mg of the target product as a white solid, which was directly used in the next step with a yield of 91.5%.

LCMS: m/z 380.16 [M+H] ⁺ .

Step 11: Preparation of (R)-5-((S)-2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-1-(4-chlorophenyl)-2-oxoethyl)-6-azaspiro[3.4]octane-6-carboxylic acid tert-butyl ester (8k)

At room temperature, compound 1b (90 mg, crude product) and dichloromethane (5 mL) were added to a reaction flask, N,N-diisopropylethylamine (427 mg, 3.31 mmol) was added, and the mixture was stirred for 5 minutes. (S)-2-((R)-6-(tert-butoxycarbonyl)-6-azaspiro[3.4]octan-5-yl)-2-(4-chlorophenyl)acetic acid (112.9 mg, 0.30 mmol) was added, and the mixture was stirred for 1 minute. Benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate (151 mg, 0.40 mmol) was added, and the mixture was stirred for 12 hours. After the reaction was completed, the mixture was concentrated under reduced pressure, and the residue was separated and purified by column chromatography (DCM:MeOH=10:1) to obtain 185 mg of the crude title compound as a white solid.

LCMS: m/z 565.26 [M+H] ⁺ .

Step 12: Preparation of (S)-1-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-2-(4-chlorophenyl)-2-((R)-6-azaspiro[3.4]octan-5-yl)ethan-1-one (8).

At room temperature, compound 8k (185 mg, crude product) and dichloromethane (3 mL) were added to a reaction flask, cooled to about 5°C in an ice bath, and a solution of hydrogen chloride in 1,4-dioxane (0.33 mL, 1.32 mmol) was added dropwise. The reaction was allowed to react at room temperature for 10 hours. After the reaction was completed, the mixture was directly concentrated under reduced pressure, and the residue was separated by preparative liquid chromatography (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um, 100 A, mobile phase: acetonitrile/water, gradient: 30%-80%) to obtain 52 mg of the title compound as a white solid, with a three-step yield of 34.0%.

LCMS: m/z 465.1 [M+H] ⁺ .

¹ H NMR (400MHz, CDCl3) δ = 10.63 (s, 1H), 8.30 (d, J = 5.9, 1H), 7.42 (d, J = 8.5, 2H), 7.34 (d, J = 8.5, 2H), 7.07(d,J＝3.6,1H),6.42(d,J＝3.7,1H),4.18-4.04(m,2H),4.04-3.95(m,1H),3.86(d,J＝13.0,1H) ,3.80 -3.54(m,5H),3.44-3.35(m,1H),2.98(dt,J＝10.7,6.2,2H),2.20(dd,J＝20.3,9.3,1H),2.14-2.06(m,1H ),1.96-1.76(m,2H),1.67-1.57(m,1H),1.53-1.40(m,2H),1.13(dd,J＝20.9,9.7,1H).

Example 9: Preparation of (S)-1-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-2-(4-chlorophenyl)-2-((S)-5-azaspiro[2.4]heptane-6-yl)ethan-1-one (9)

Step 1: Preparation of methyl 2-cyclopropylidene acetate (9a)

A solution of (1-ethoxycyclopropyloxy)trimethylsilane (20.0 g, 114.8 mmol) in MeOH (150 mL) was stirred at 25°C for 1 hour, then concentrated under reduced pressure, and the residue was dissolved in toluene (500 mL). Benzoic acid (7.0 g, 57.4 mmol) was added to the above solution. The mixture was heated to 90°C with stirring, and then a solution of ethoxycarbonylmethylenetriphenylphosphine (40 g, 114.8 mmol) in toluene (1200 mL) was added at the same temperature. The obtained mixture was stirred at 90°C for 16 hours, and then concentrated under reduced pressure. The residue was purified by column chromatography (PE:EA=10:1) to obtain 8.5 g of the title compound, with a yield of 66.4%.

LCMS: m/z 113.05 [M+H] ⁺ .

Step 2: Preparation of methyl 2-(1-(nitromethyl)cyclopropyl)acetate (9b)

At room temperature, methyl 2-cyclopropylidene acetate (9a) (8.5 g, 75.8 mol) was added to dimethyl sulfoxide (50 mL), and then cesium carbonate (12.3 g, 37.9 mol) and nitromethane (8.75 g, 113.7 mol) were added in sequence, and stirred at 80°C overnight. The reaction solution was quenched with glacial acetic acid (2.04 g, 0.340 mol), extracted with methyl tert-butyl ether, the organic phase was dried, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: EA/PE = 0:1-1:10) to obtain 3.8 g of the title compound, yield: 29.0%.

LCMS: m/z 174.07 [M+H] ⁺ .

Step 3: Preparation of 5-azaspiro[2.4]heptan-6-one (9c)

At room temperature, methyl 2-(1-(nitromethyl)cyclopropyl)acetate (9b) (3.8 g, 21.9 mmol) was added to ethyl acetate (80 mL), and then Pd/C (1.1 g) was added. The mixture was stirred overnight at room temperature under a hydrogen atmosphere. The reaction solution was filtered, the organic phase was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: MeOH/DCM = 0:1-1:50) to obtain 1.5 g of the title compound, with a yield of 62.5%.

LCMS: m/z 112.07 [M+H] ⁺ .

Step 4: Preparation of tert-butyl 6-oxo-5-azaspiro[2.4]heptane-5-carboxylate (9d)

At room temperature, 5-azaspiro[2.4]heptane-6-one (9c) (1.5 g, 13.5 mmol) was added to dichloromethane (50 mL), followed by di-tert-butyl dicarbonate (4.4 g, 20.2 mmol), triethylamine (4.1 g, 40.5 mmol) and 4-dimethylaminopyridine (82 mg, 0.675 mmol). The mixture was stirred at room temperature for 2 hours. The reaction solution was extracted with dichloromethane, the organic phase was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: methyl tert-butyl ether (MTBE)/DCM = 0:1-1:10) to obtain 1.4 g of the title compound, yield: 49.1%.

LCMS: m/z 156.12 [M-55] ⁺ .

Step 5: Preparation of tert-butyl 6-hydroxy-5-azaspiro[2.4]heptane-5-carboxylate (9e)

At room temperature, tert-butyl 6-oxo-5-azaspiro[2.4]heptane-5-carboxylate (9d) (1.4 g, 6.63 mmol) and THF (30 mL) were added to the reaction flask. The temperature was lowered to -78 °C, and DIBAL-H (10 mL, 10.0 mmol) was added dropwise. After the addition was completed, the mixture was stirred at -78 °C for 30 minutes. After the reaction was completed, the reaction was quenched with saturated aqueous sodium acetate solution (5 mL), warmed to room temperature, and saturated aqueous ammonium chloride solution (5 mL) and ethyl acetate (10 mL) were added and stirred for 10 minutes. The mixture was filtered, and the aqueous phase was extracted with ethyl acetate. After the organic phases were combined, the mixture was concentrated under reduced pressure to obtain 1.27 g of the title product as a yellow oil, which was used directly in the next step with a yield of 90.0%.

LCMS: m/z 214.14 [M+H] ⁺ .

Step 6: Preparation of tert-butyl 6-methoxy-5-azaspiro[2.4]heptane-5-carboxylate (9f)

At room temperature, tert-butyl 6-hydroxy-5-azaspiro[2.4]heptane-5-carboxylate (9e) (1.27 g, 5.95 mmol), methanol (20 mL) and p-toluenesulfonic acid monohydrate (52.0 mg, 0.270 mmol) were added to a reaction flask and reacted at room temperature for 2 hours. After the reaction, the reaction system was poured into a saturated sodium bicarbonate aqueous solution and extracted with ethyl acetate. The organic phases were combined and concentrated under reduced pressure. The residue was diluted with dichloromethane to a 29 wt% solution and used directly in the next step.

LCMS: m/z 228.15 [M+H] ⁺ .

Step 7: Preparation of (S)-tert-butyl 6-((S)-2-((R)-4-benzyl-2-oxooxazolidin-3-yl)-1-(4-chlorophenyl)-2-oxoethyl)-5-azaspiro[2.4]heptane-5-carboxylate (9 g)

At room temperature, compound 1i (1.86 g, 5.64 mmol) and dichloromethane (30 mL) were added to a reaction flask, cooled to -78°C, titanium tetrachloride (1.18 g, 6.23 mmol) was added dropwise, and DIPEA (878 mg, 6.79 mmol) was added dropwise after 30 minutes of incubation. After 30 minutes of incubation, a dichloromethane solution of 6-methoxy-5-azaspiro[2.4]heptane-5-carboxylic acid tert-butyl ester (9f) was added dropwise, and the temperature was naturally raised to room temperature for 1.5 hours of reaction. After the reaction was completed, the reaction system was poured into water, the aqueous phase was extracted with dichloromethane, the organic phases were combined, dried over anhydrous Na ₂ SO ₄ , filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by column chromatography (PE:EA=5:1) to obtain 1.76 g of the target product as a yellow oil, which was directly used in the next step with a yield of 59.6%.

LCMS: m/z 525.21 [M+H] ⁺ .

Step 8: Preparation of (S)-2-((S)-5-(tert-butyloxycarbonyl)-5-azaspiro[2.4]heptane-6-yl)-2-(4-chlorophenyl)acetic acid (9h)

At room temperature, LiOH (1.40 g, 33.5 mmol), water (20 mL) and THF (40 mL) were added to the reaction bottle. The mixture was cooled to about 5°C in an ice bath, 30% hydrogen peroxide (569 mg, 5.0 mmol) was added dropwise, and (S)-6-((S)-2-((R)-4-benzyl-2-oxooxazolidin-3-yl)-1-(4-chlorophenyl)-2-oxoethyl)-5-azaspiro[2.4]heptane-5-carboxylic acid tert-butyl ester (1.76 g, 3.35 mmol) was added in batches, and the mixture was reacted in an ice bath for 30 hours. After the reaction was completed, the reaction was quenched with a 12.5 wt % aqueous sodium sulfite solution, the pH was adjusted to 2 with a 27.0 wt % potassium bisulfate solution, the aqueous phase was extracted with ethyl acetate, the organic phases were combined and concentrated under reduced pressure, and the residue was separated and purified by column chromatography (PE:EA=10:1) to obtain 896 mg of the target product as a yellow oil, which was directly used in the next step with a yield of 73.1%.

LCMS: m/z 366.14 [M+H] ⁺ .

Step 9: Preparation of (S)-tert-butyl 6-((S)-2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-1-(4-chlorophenyl)-2-oxoethyl)-5-azaspiro[2.4]heptane-5-carboxylate (9i).

At room temperature, compound 1b (90 mg, crude product) and dichloromethane (5 mL) were added to a reaction flask, N,N-diisopropylethylamine (427 mg, 3.31 mmol) was added, and the mixture was stirred for 5 minutes. (S)-2-((S)-5-(tert-butyloxycarbonyl)-5-azaspiro[2.4]heptane-6-yl)-2-(4-chlorophenyl)acetic acid (112.9 mg, 0.31 mmol) was added, and the mixture was stirred for 1 minute. Benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate (151 mg, 0.40 mmol) was added, and the mixture was stirred for 12 hours. After the reaction was completed, the mixture was concentrated under reduced pressure, and the residue was separated and purified by column chromatography (DCM:MeOH=10:1) to obtain 250 mg of the crude title compound as a white solid.

LCMS: m/z 551.25 [M+H] ⁺ .

Step 10: Preparation of (S)-1-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-2-(4-chlorophenyl)-2-((S)-5-azaspiro[2.4]heptan-6-yl)ethan-1-one (9).

At room temperature, compound 9i (250 mg, crude product) and dichloromethane (3 mL) were added to a reaction flask, cooled to about 5°C in an ice bath, and a solution of hydrogen chloride in 1,4-dioxane (0.33 mL, 1.32 mmol) was added dropwise. The reaction was allowed to react at room temperature for 10 hours. After the reaction was completed, the mixture was directly concentrated, and the residue was separated by preparative liquid chromatography (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um, 100 A, mobile phase: acetonitrile/water, gradient: 30%-80%) to obtain 91 mg of the title compound as a white solid, with a three-step yield of 61.2%.

LCMS: m/z 451.10 [M+H] ⁺ .

¹ H NMR (400MHz, CDCl3) δ = 10.63 (s, 1H), 8.27 (s, 1H), 7.35-7.28 (m, 2H), 7.24 (s, 2H), 7.08 (d, J = 3.6, 1H) ,6.41(d,J＝3.7,1H),4.13-4.03(m,1H),3.99(ddd,J＝16.7,11.5,6.4,3H),3.93-3.85(m,1H),3.76 -3.67(m,1H),3.60(ddd,J＝14.4,7.5,3.7,2H),3.50-3.41(m,1H),3.36-3.27(m,1H),3.01(q,J＝9.8,2H ),1.79-1.67(m,1H),1.29-1.18(m,1H),0.68-0.58(m,1H),0.58-0.51(m,1H),0.51-0.40(m,2H).

Example 10: Preparation of (S)-1-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-2-(4-chlorophenyl)-2-((S)-6-azaspiro[3.4]octan-7-yl)ethan-1-one (10)

Step 1: Preparation of methyl 2-cyclobutylidene acetate (10a)

Dissolve methyl 2-(diethoxyphosphoryl)acetate (8.97 g, 42.7 mmol) in anhydrous tetrahydrofuran (80 mL), slowly add sodium hydride (2.28 g, 57.0 mmol) in batches under ice bath conditions, and react at room temperature for 1 hour. Then add cyclobutanone (2.0 g, 28.5 mmol) and stir at room temperature for 4 hours. The reaction solution was concentrated under reduced pressure, the residue was extracted with ethyl acetate and water, the organic phase was dried and concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: EA/PE = 0:1-1:10) to obtain 2.6 g of the oily title compound, yield: 72.2%.

LCMS: m/z 127.07 [M+H] ⁺ .

Step 2: Preparation of methyl 2-(1-(nitromethyl)cyclobutyl)acetate (10b)

At room temperature, methyl 2-cyclobutylidene acetate (10a) (2.6 g, 20.6 mmol) was added to dimethyl sulfoxide (50 mL), and then cesium carbonate (3.35 g, 10.3 mmol) and nitromethane (7.33 g, 1.19 mol) were added in sequence, and stirred at 80°C overnight. The reaction solution was quenched with glacial acetic acid (2.04 g, 0.340 mol), extracted with methyl tert-butyl ether, the organic phase was dried, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: EA/PE = 0:1-1:10) to obtain 1.4 g of the title compound, yield: 36.3%.

LCMS: m/z 188.08 [M+H] ⁺ .

Step 3: Preparation of 6-azaspiro[3.4]octan-7-one (10c)

At room temperature, methyl 2-(1-(nitromethyl)cyclobutyl)acetate (10b) (1.4 g, 7.48 mmol) was added to ethyl acetate (50 mL), and then Pd/C catalyst (420 mg) was added. The mixture was stirred overnight at room temperature under a hydrogen atmosphere. The reaction solution was filtered, the organic phase was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: MeOH/DCM = 0:1-1:50) to obtain 1.1 g of the crude title compound, which was used directly in the next step.

LCMS: m/z 126.08 [M+H] ⁺ .

Step 4: Preparation of tert-butyl 7-oxo-6-azaspiro[3.4]octane-6-carboxylate (10d)

At room temperature, 6-azaspiro[3.4]octan-7-one (10c) (1.1 g, 8.79 mmol) was added to dichloromethane (50 mL), followed by di-tert-butyl dicarbonate (2.88 g, 13.2 mmol), triethylamine (TEA) (2.67 g, 26.4 mmol) and 4-dimethylaminopyridine (DMAP) (53.7 mg, 0.44 mmol), and stirred at room temperature for 2 hours. The reaction solution was extracted with dichloromethane, the organic phase was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: MTBE/DCM = 0:1-1:10) to obtain 1.2 g of the title compound, yield: 60.3%.

LCMS: m/z 170.14 [M-55] ⁺ .

Step 5: Preparation of tert-butyl 7-hydroxy-6-azaspiro[3.4]octane-6-carboxylate (10e)

At room temperature, tert-butyl 7-oxo-6-azaspiro[3.4]octane-6-carboxylate (10d) (1.2 g, 5.33 mmol) and THF (24 mL) were added to the reaction flask. The temperature was lowered to -78 °C, and DIBAL-H (10 mL, 10.0 mmol) was added dropwise. After the addition was completed, the mixture was stirred at -78 °C for 30 minutes. After the reaction was completed, the reaction was quenched with saturated aqueous sodium acetate solution (5 mL), warmed to room temperature, and saturated aqueous ammonium chloride solution (5 mL) and ethyl acetate (10 mL) were added and stirred for 10 minutes. The mixture was filtered, and the aqueous phase was extracted with ethyl acetate. After the organic phases were combined, the mixture was concentrated under reduced pressure to obtain 1.11 g of the title product as a yellow oil, which was used directly in the next step with a yield of 91.7%.

LCMS: m/z 228.15 [M+H] ⁺ .

Step 6: Preparation of tert-butyl 7-methoxy-6-azaspiro[3.4]octane-6-carboxylate (10f)

At room temperature, tert-butyl 7-hydroxy-6-azaspiro[3.4]octane-6-carboxylate (10e) (1.11 g, 4.89 mmol), methanol (20 mL) and p-toluenesulfonic acid monohydrate (52.0 mg, 0.270 mmol) were added to a reaction flask and reacted at room temperature for 2 hours. After the reaction, the reaction system was poured into a saturated sodium bicarbonate aqueous solution and extracted with ethyl acetate. The organic phases were combined and concentrated under reduced pressure. The residue was diluted with dichloromethane to a 29 wt% solution and used directly in the next step.

LCMS: m/z 242.17 [M+H] ⁺ .

Step 7: Preparation of (S)-tert-butyl 7-((S)-2-((R)-4-benzyl-2-oxooxazolidin-3-yl)-1-(4-chlorophenyl)-2-oxoethyl)-6-azaspiro[3.4]octane-6-carboxylate (10 g)

At room temperature, compound 1i (1.86 g, 5.64 mmol) and dichloromethane (30 mL) were added to the reaction flask. The temperature was lowered to -78 °C, titanium tetrachloride (1.18 g, 6.23 mmol) was added dropwise, and DIPEA (878 mg, 6.79 mmol) was added dropwise after 30 minutes of incubation. After 30 minutes of incubation, a dichloromethane solution of tert-butyl 7-methoxy-6-azaspiro[3.4]octane-6-carboxylate (10f) was added dropwise, and the temperature was naturally raised to room temperature for 1.5 hours of reaction. After the reaction was completed, the reaction system was poured into water, the aqueous phase was extracted with dichloromethane, the organic phases were combined, dried over anhydrous Na ₂ SO ₄ , filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by column chromatography (PE:EA=5:1) to obtain 1.5 g of the target product as a yellow oil, which was directly used in the next step with a yield of 57.0%.

LCMS: m/z 539.22 [M+H] ⁺ .

Step 8: Preparation of (S)-2-((S)-6-(tert-butyloxycarbonyl)-6-azaspiro[3.4]octan-7-yl)-2-(4-chlorophenyl)acetic acid (10h)

At room temperature, LiOH (175 mg, 4.18 mmol), water (20 mL), and THF (40 mL) were added to the reaction flask. The mixture was cooled to about 5°C in an ice bath, 30% hydrogen peroxide (472 mg, 4.16 mmol) was added dropwise, and (S)-7-((S)-2-((S)-4-benzyl-2-oxooxazolidin-3-yl)-1-(4-chlorophenyl)-2-oxoethyl)-6-azaspiro[3.4]octane-6-carboxylic acid tert-butyl ester (10 g) (1.5 g, 2.78 mmol) was added in batches, and the mixture was reacted in an ice bath for 30 hours. After the reaction, the reaction was quenched with a 12.5 wt % aqueous sodium sulfite solution, the pH was adjusted to 2 with a 27.0 wt % potassium bisulfate solution, the aqueous phase was extracted with ethyl acetate, the organic phases were combined and concentrated under reduced pressure, and the residue was separated by column chromatography (PE:EA=10:1) to obtain 777 mg of the target product as a yellow oil, which was directly used in the next step with a yield of 73.5%.

LCMS: m/z 380.16 [M+H] ⁺ .

Step 9: Preparation of (S)-tert-butyl 7-((S)-2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-1-(4-chlorophenyl)-2-oxoethyl)-6-azaspiro[3.4]octane-6-carboxylate (10i).

At room temperature, compound 1b (90 mg, crude product) and dichloromethane (5 mL) were added to a reaction flask, N,N-diisopropylethylamine (427 mg, 3.31 mmol) was added, and the mixture was stirred for 5 minutes. (S)-2-((S)-6-(tert-butyloxycarbonyl)-6-azaspiro[3.4]octan-7-yl)-2-(4-chlorophenyl)acetic acid (112.9 mg, 0.30 mmol) was added, and the mixture was stirred for 1 minute. Benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate (151 mg, 0.40 mmol) was added, and the mixture was stirred for 12 hours. After the reaction was completed, the mixture was concentrated under reduced pressure, and the residue was separated and purified by column chromatography (eluent) to obtain 250 mg of the crude title compound as a white solid.

LCMS: m/z 565.26 [M+H] ⁺ .

Step 10: Preparation of (S)-1-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-2-(4-chlorophenyl)-2-((S)-6-azaspiro[3.4]octan-7-yl)ethan-1-one (10).

At room temperature, compound 10i (250 mg, crude product) and dichloromethane (3 mL) were added to a reaction flask, cooled to about 5°C in an ice bath, and a solution of hydrogen chloride in 1,4-dioxane (0.33 mL, 1.32 mmol) was added dropwise. The reaction was allowed to react at room temperature for 10 hours. After the reaction was completed, the mixture was directly concentrated, and the residue was separated by preparative liquid chromatography (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um, 100 A, mobile phase: acetonitrile/water, gradient: 30%-80%) to obtain 89 mg of the title compound as a white solid, with a three-step yield of 58.1%.

LCMS: m/z 465.10 [M+H] ⁺ .

¹ H NMR (400MHz, CDCl3) δ = 10.77 (s, 1H), 8.27 (s, 1H), 7.39-7.29 (m, 2H), 7.26-7.22 (m, 2H), 7.07 (d, J = 3.6, 1H),6.39(d,J＝3.7,1H),4.03-3.93(m,2H),3.89(dd,J＝14.2,6.3,3H),3.68(ddd,J＝12.4,6.6,2.6,1H) ,3.61-3.49(m,2H),3.47-3.38(m,1H),3.32-3.21(m,1H),3.07(q,J＝10.6,2H),2.09-1.99(m,1H),1.99 -1.88(m,2H),1.85-1.75(m,3H),1.61-1.48(m,2H).

Biological tests

Test Example 1: Inhibitory activity of the compounds of the present invention on AKT1/AKT2/AKT3 kinases

The experiment used the HTRF method to detect the inhibitory activity of the compounds on AKT kinase.

Test methods

Materials and reagents: AKT1, AKT2, and AKT3 were purchased from Thermo, with catalog numbers P2999, PV3184, and PV3185, respectively. HTRF KinEASE-STK S3 kit was from Cisbio, with catalog number 62ST3PEB. Other reagents: ATP solution (Sigma, R0441), MgCl ₂ (Sigma, 7786-30-3), DMSO (Sigma, D2650-100ML), and Ipatasertib (MCE, HY-15186).

Experimental procedure: First, dilute the 5× kinase buffer to prepare a 1× experimental working solution, with final concentrations of MgCl ₂ and DTT of 5mM and 1mM, respectively. The compound dilution was performed according to the 27-fold dilution method, and a total of 4 concentration points were diluted from the highest concentration point, and transferred to the Echo plate (Labcyte, LP0200). The compound was transferred to a 384-well plate (Greiner, 784075) using Echo to make the compound a 3-fold dilution matrix with 11 concentration points. Add 5ul of 2× kinase (final concentration AKT1: 0.02ng/ul; AKT2: 0.03ng/ul; AKT3: 0.01ng/ul) to the 384-well plate, centrifuge at 1000rpm for 30 seconds, and incubate at room temperature for 15 minutes. Add 5ul of 2×STK-Sub3-biotin substrate and ATP (final concentration AKT1: 150uM; AKT2: 250uM; AKT3: 100uM) working solution to a 384-well plate, centrifuge at 1000rpm for 30 seconds, and incubate at room temperature for 75 minutes. Add 10ul of detection solution to a 384-well plate, centrifuge at 1000rpm for 30 seconds, and incubate at room temperature for 30 minutes. Envision microplate reader (PerkinElmer) detects 665/615 fluorescence signal values. XL-Fit software was used for data analysis, and compound IC ₅₀ was obtained by nonlinear fitting.

The IC ₅₀ values of the compounds of the present invention for inhibiting AKT1, AKT2 and AKT3 kinases are shown in Table 1 below.

Table 1 IC ₅₀ values of the compounds of the present invention for inhibition of AKT1, AKT2 and AKT3 kinases

Conclusion: As can be seen from Table 1, the compounds of the present invention have a significant inhibitory effect on AKT1/2/3 kinases.

Test Example 2: Inhibitory effect of the compounds of the present invention on prostate cancer cells

The CTG method was used to detect the inhibitory level of the compounds of the present invention on prostate cancer Lncap cells (PTEN deficiency).

LNCaP cells (ATCC, CRL-1740) were cultured in RPMI-1640 medium (Gibco, C11875500BT) supplemented with 10% FBS (Gibco, 10099141) and double antibody (1% penicillin and streptomycin, Gibco, 15140122). When the cells grew to 70-80% confluence, they were digested for 8 minutes using the pancreatic enzyme substitute Tryple ^TM Express (Invitrogen, 12604021). Cell suspension was prepared in RPMI-1640 culture medium supplemented with 10% FBS, counted, and the cell density was adjusted to 1.25×10 ⁴ /ml. The cell suspension was added to a 384-well plate, 40 μl per well, i.e., 500 LNCaP cells/well. Compound preparation: The compound was diluted 3 times in 100% DMSO starting from 10 mM, with 10 concentration gradients. Then, the medium containing 10% FBS was used for intermediate dilution, and 1 μl of the compound was added to 199 μl of the medium, and diluted 200 times. The starting concentration of the experiment was 10 μM. 10 μl of the compound diluted in the medium was added to the cells in the 384-well plate, 2 replicate wells, a total of 50 μl reaction system, and the final concentration of DMSO was 0.1%. Cultured in a 37°C, 5% CO ₂ incubator for 96 hours. Then the 384-well plate was taken out, equilibrated at room temperature for 30 minutes, 20 μl Cell Titer-Glo (Promega, Cat. No.: G7572) was added to each well, oscillated and mixed, and incubated at room temperature for 10 minutes. The luminescence value was read by a multifunctional microplate reader (Biotek, model Cytation 3). GraphPad prism 8.0 software was used for nonlinear fitting to calculate the IC ₅₀ value of the compound's inhibition of cell proliferation.

The IC ₅₀ values of the compounds of the present invention for inhibiting LNCaP cells are shown in Table 2 below.

Table 2 IC ₅₀ values of the compounds of the present invention for inhibition of LNCaP cells

Compound No. IC ₅₀ (nM) 1 96.79 2 107.25 3 23.00 4 32.00 5 65.57 6 112.78 7 109.45 8 62.39 9 35.43 10 69.95

Conclusion: It can be seen from Table 2 that the compounds of the present invention have a significant inhibitory effect on the proliferation of human prostate cancer LNCaP cells.

Claims (21)

1. A compound represented by general formula (I) or its stereoisomers, tautomers, mesoforms, racemates, enantiomers, diastereoisomers, or its mixture form, or its pharmaceutically acceptable salt, in: Ring A is selected from C ₃ -C ₁₂ cycloalkyl, 3 to 14 membered heterocyclic group, C ₆ -C ₁₄ aryl and 6 to 14 membered heteroaryl; wherein the C ₃ -C ₁₂ cycloalkyl, 3 to 14-membered heterocyclyl, C ₆ -C ₁₄ aryl, and 6 to 14-membered heteroaryl are each independently optionally substituted by one or more R ¹ ; L is a bond or -CH ₂ -; Ring B is selected from 5 to 8 membered saturated heterocyclic groups containing 1-2 nitrogen atoms; the heterocyclic group is optionally substituted by one or more R ³ ; G is selected from hydrogen, phenyl, 5-6 membered heteroaryl, wherein the phenyl, 5-6 membered heteroaryl is optionally substituted by one or more R ⁴ ; Z is CH or N; Each R ^is independently selected from hydrogen, halogen, amino, nitro, cyano, oxo, hydroxyl, mercapto, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, Aryl, heteroaryl, said alkyl, alkoxyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl are optionally selected from halogen, amino, nitro, cyano , hydroxyl, mercapto, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl or multiple groups substituted; or, Any two ^R together with the atoms to which they are attached form cycloalkyl, heterocyclyl, aryl and heteroaryl which are optionally selected from the group consisting of halogen, heterocyclyl, aryl and heteroaryl Amino, nitro, cyano, oxo, hydroxyl, mercapto, carboxyl, ester, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl One or more groups of radicals and heteroaryls are substituted; Each ^{R is} independently selected from hydrogen, halogen, amino, nitro, cyano, oxo, hydroxyl, mercapto, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, Aryl, heteroaryl, said alkyl, alkoxyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl are optionally selected from halogen, amino, nitro, cyano , hydroxyl, mercapto, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl or multiple groups substituted; or, Any two R ² together with the atoms to which they are attached form cycloalkyl, heterocyclyl, aryl and heteroaryl, which are optionally selected from the group consisting of halogen, heterocyclyl, aryl and heteroaryl Amino, nitro, cyano, oxo, hydroxyl, mercapto, carboxyl, ester, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl One or more groups of radicals and heteroaryls are substituted; Each ^R is independently selected from hydrogen, halogen, amino, nitro, cyano, oxo, hydroxyl, mercapto, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, Aryl, heteroaryl, said alkyl, alkoxyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl are optionally selected from halogen, amino, nitro, cyano , hydroxyl, mercapto, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl or multiple groups substituted; or, Any two R ³ together with the atoms to which they are attached form cycloalkyl, heterocyclyl, aryl and heteroaryl, which are optionally selected from the group consisting of halogen, heterocyclyl, aryl and heteroaryl Amino, nitro, cyano, oxo, hydroxyl, mercapto, carboxyl, ester, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl One or more groups of radicals and heteroaryls are substituted; ^Each R is independently selected from the group consisting of hydrogen, halogen, nitro, cyano, hydroxyl, mercapto, alkyl, alkenyl, alkynyl, -S(O) _p R ^a , -NR ^a R ^b , -OR ^a , -C(O)R ^a , -C(O)NR ^a R ^b , -S(O) _p NR ^a R ^b , cycloalkyl, heterocyclyl, aryl, heteroaryl, the alkyl, Alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl are optionally selected from halogen, amino, nitro, cyano, hydroxyl, mercapto, carboxyl, ester, oxo, alkyl , alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl are substituted by one or more groups; R ^a and R ^b are each independently selected from hydrogen, halogen, hydroxyl, mercapto, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, said alkyl, alkenyl, Alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl are optionally selected from halogen, amino, nitro, cyano, hydroxyl, mercapto, carboxyl, ester, oxo, alkyl, alkoxy One or more groups of radical, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl; or, R ^a and R ^b form a nitrogen-containing heterocyclic group together with the nitrogen atom to which they are attached, and the nitrogen-containing heterocyclic group is optionally selected from the group consisting of halogen, amino, nitro, cyano, oxo, hydroxyl, mercapto, carboxyl , ester group, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are substituted by one or more groups; n is 0, 1, 2 or 3; p is 0, 1 or 2. 2. The compound represented by the general formula (I) according to claim 1 or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer Isomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, wherein ring A is selected from C ₃ -C ₁₂ cycloalkyl and 3 to 14 membered heterocyclic groups, preferably 3 to 14 membered heterocyclic groups, more preferably 3 to 6 membered monocyclic heterocyclic group, 6 to 11 membered bicyclic heterocyclic group, more preferably 6 to 10 membered condensed heterocyclic group, 6 to 10 membered spiro heterocyclic group; the cycloalkyl or heterocyclic group are independently is optionally substituted by one or more R ¹ ; Each R ¹ is independently selected from hydrogen, halogen, amino, nitro, cyano, oxo, hydroxyl, mercapto, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclic group, C ₆ -C ₁₀ aryl, 5 to 10 membered heteroaryl, the C ₁ - C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclyl, C ₆ - C ₁₀ aryl, 5 to 10 membered heteroaryl is optionally selected from halogen, amino, nitro, cyano, hydroxyl, mercapto, carboxyl, ester, oxo, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6-membered heterocyclic group, C ₆ -C ₁₀ aryl group, 5 to 10-membered heteroaryl group substituted by one or more groups; or, Any two ^R together with the atoms to which they are attached form cycloalkyl, heterocyclyl, aryl and heteroaryl which are optionally selected from the group consisting of halogen, heterocyclyl, aryl and heteroaryl Amino, nitro, cyano, oxo, hydroxyl, mercapto, carboxyl, ester, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclyl, C ₆ -C ₁₀ aryl and 5 to 10 membered One or more groups of heteroaryl are substituted. 3. according to the compound shown in the general formula (I) described in claim 1 or 2 or its stereoisomer, tautomer, mesoform, racemate, enantiomer, non- An enantiomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, which is a compound represented by general formula (II) or its stereoisomer, tautomer, meso, racemic isomers, enantiomers, diastereoisomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, in: m is 0, 1, 2, 3 or 4; Each R ¹ is independently selected from hydrogen, halogen, amino, nitro, cyano, oxo, hydroxyl, mercapto, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclic group, C ₆ -C ₁₀ aryl, 5 to 10 membered heteroaryl, the C ₁ - C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclyl, C ₆ - C ₁₀ aryl, 5 to 10 membered heteroaryl is optionally selected from halogen, amino, nitro, cyano, hydroxyl, mercapto, carboxyl, ester, oxo, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkoxy, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclyl, C ₆ -C ₁₀ aryl, 5 to 10-membered heteroaryl by one or more groups; or, Any two ^R together with the atoms to which they are attached form cycloalkyl, heterocyclyl, aryl and heteroaryl which are optionally further selected from the group consisting of halogen , amino, nitro, cyano, oxo, hydroxyl, mercapto, carboxyl, ester, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ - C ₆ haloalkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclyl, C ₆ -C ₁₀ aryl and 5 to 10 Substituted by one or more groups of heteroaryl; Rings B, L, G, Z, R ² , n are as defined in claim 1 . 4. The compound represented by the general formula (I) according to any one of claims 1 to 3 or its stereoisomers, tautomers, mesomers, racemates, enantiomers isomers, diastereoisomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, wherein any two R ¹ form C ₃ -C ₆ cycloalkyl or 3 to 6-membered heterocycle together with the atoms they are connected to The C ₃ -C ₆ cycloalkyl or 3 to 6 membered heterocyclic group is optionally selected from halogen, amino, nitro, cyano, oxo, hydroxyl, mercapto, carboxyl, ester, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclic group, C ₆ -C ₁₀ aryl and 5 to 10 membered heteroaryl are substituted by one or more groups. 5. The compound represented by the general formula (I) according to any one of claims 1 to 4 or its stereoisomers, tautomers, mesomers, racemates, enantiomers isomers, diastereoisomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, wherein L is a bond. 6. The compound represented by the general formula (I) according to any one of claims 1 to 5 or its stereoisomers, tautomers, mesomers, racemates, enantiomers isomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, which is represented by general formula (III-1), general formula (III-2) or general formula (III-3) A compound or its stereoisomers, tautomers, mesoforms, racemates, enantiomers, diastereoisomers, or a mixture thereof, or a pharmaceutically acceptable salt thereof, in, X ₁ and X ₂ are each independently selected from -CH ₂ -, -O-, -NH-; n ₁ is 0 or 1; _n2 is 0, 1 or 2; Each R ^c is independently selected from halogen, amino, nitro, cyano, oxo, hydroxyl, mercapto, carboxyl, ester, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclyl, C ₆ -C ₁₀ aryl and 5 to 10 membered heteroaryl; q is 0, 1 or 2, preferably 0 or 1, more preferably 0; Rings B, G, Z, ^R2 and n are as defined in claim 1. 7. The compound represented by the general formula (I) according to any one of claims 1 to 5 or its stereoisomers, tautomers, mesomers, racemates, enantiomers isomers, diastereoisomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, which are compounds represented by general formula (IV-1) or (IV-2) or stereoisomers, interconversions thereof Isomers, mesoforms, racemates, enantiomers, diastereoisomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, in, Ring E ₁ is C ₃ -C ₆ cycloalkyl or 3 to 6 membered heterocyclic group; Ring E ₂ is C ₃ -C ₆ cycloalkyl or 3 to 6 membered heterocyclic group; Each R ^e is independently selected from halogen, amino, nitro, cyano, oxo, hydroxyl, mercapto, carboxyl, ester, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclyl, C ₆ -C ₁₀ aryl and 5 to 10 membered heteroaryl; t is 0, 1 or 2, preferably 0 or 1, more preferably 0; Rings B, G, Z, ^R2 and n are as defined in claim 1. 8. The compound represented by the general formula (I) according to any one of claims 1 to 7 or its stereoisomers, tautomers, mesomers, racemates, enantiomers isomer, diastereoisomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, wherein ring B is a 6-8 membered saturated heterocyclic group containing 1-2 nitrogen atoms; ring B is preferably selected from the following groups group: Wherein the single wave line represents the position connected to the carbonyl, and the double wave line represents the position connected to the pyrimidine; Ring B is optionally substituted by one or more ^R3 ; Each R ³ is independently selected from hydrogen, halogen, amino, nitro, cyano, oxo, hydroxyl, mercapto, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclic group, C ₆ -C ₁₀ aryl, 5 to 10 membered heteroaryl, the C ₁ - C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclyl, C ₆ - C ₁₀ aryl, 5 to 10 membered heteroaryl is optionally selected from halogen, amino, nitro, cyano, hydroxyl, mercapto, carboxyl, ester, oxo, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 Substituted by one or more groups from 6-membered heterocyclic group, C ₆ -C ₁₀ aryl group, 5- to 10-membered heteroaryl group; Preferably ^R3 is hydrogen. 9. according to the compound shown in the general formula (I) described in any one in claim 1 to 6 or 8 or its stereoisomer, tautomer, mesoform, racemate, para Enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, which are compounds represented by general formula (IIIA), general formula (IIIB) or general formula (IIIC) or their stereo Isomers, tautomers, mesoforms, racemates, enantiomers, diastereoisomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, in, X ₁ and X ₂ are each independently selected from -CH ₂ -, -O-, -NH-; n ₁ is 0 or 1; _n2 is 0, 1 or 2; Each R ^c is independently selected from halogen, amino, nitro, cyano, oxo, hydroxyl, mercapto, carboxyl, ester, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclyl, C ₆ -C ₁₀ aryl and 5 to 10 membered heteroaryl; q is 0, 1 or 2, preferably 0 or 1, more preferably 0; Each R ³ is independently selected from hydrogen, halogen, amino, nitro, cyano, oxo, hydroxyl, mercapto, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclic group, C ₆ -C ₁₀ aryl, 5 to 10 membered heteroaryl, the C ₁ - C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclyl, C ₆ - C ₁₀ aryl, 5 to 10 membered heteroaryl is optionally selected from halogen, amino, nitro, cyano, hydroxyl, mercapto, carboxyl, ester, oxo, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6-membered heterocyclic group, C ₆ -C ₁₀ aryl group, 5 to 10-membered heteroaryl group substituted by one or more groups; preferably R ³ is hydrogen; r is 0, 1, 2, 3 or 4, preferably 0, 1 or 2, more preferably 0 or 1; G, Z, ^R2 and n are as defined in claim 1. 10. The compound represented by the general formula (I) according to any one of claims 1 to 5 or 7 to 8 or its stereoisomers, tautomers, mesomers, racemates , enantiomers, diastereoisomers, or a mixture thereof, or a pharmaceutically acceptable salt thereof, which is a compound represented by general formula (IVA) or (IVB) or a stereoisomer, tautomer Isomers, mesoforms, racemates, enantiomers, diastereoisomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, in, Ring E ₁ is C ₃ -C ₆ cycloalkyl or 3 to 6 membered heterocyclic group; Ring E ₂ is C ₃ -C ₆ cycloalkyl or 3 to 6 membered heterocyclic group; Each R ^e is independently selected from halogen, amino, nitro, cyano, oxo, hydroxyl, mercapto, carboxyl, ester, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclyl, C ₆ -C ₁₀ aryl and 5 to 10 membered heteroaryl; t is 0, 1 or 2, preferably 0 or 1, more preferably 0; Each R ³ is independently selected from hydrogen, halogen, amino, nitro, cyano, oxo, hydroxyl, mercapto, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclic group, C ₆ -C ₁₀ aryl, 5 to 10 membered heteroaryl, the C ₁ - C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclyl, C ₆ - C ₁₀ aryl, 5 to 10 membered heteroaryl is optionally selected from halogen, amino, nitro, cyano, hydroxyl, mercapto, carboxyl, ester, oxo, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6-membered heterocyclic group, C ₆ -C ₁₀ aryl group, 5 to 10-membered heteroaryl group substituted by one or more groups; preferably R ³ is hydrogen; r is 0, 1, 2, 3 or 4, preferably 0, 1 or 2, more preferably 0 or 1; G, Z, ^R2 and n are as defined in claim 1. 11. The compound represented by the general formula (I) according to any one of claims 1 to 10 or its stereoisomers, tautomers, mesomers, racemates, enantiomers isomer, diastereoisomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, wherein G is selected from phenyl, and the phenyl is optionally substituted by one or more R ⁴ ; Each R ⁴ is independently selected from halogen, cyano, hydroxyl, mercapto, C ₁ -C ₆ alkyl, -NR ^a R ^b , -OR ^a , said C ₁ -C ₆ alkyl being optionally substituted by halogen ; R ^a and R ^b are each independently selected from hydrogen and C ₁ -C ₆ alkyl; or, R ^a and R ^b form a 5-6 membered nitrogen-containing heterocyclic group together with their connected nitrogen atoms, and the 5-6 membered nitrogen-containing heterocyclic group is optionally selected from halogen, amino, nitro, cyano, oxygen One or more of substituent, hydroxyl, mercapto, carboxyl, ester, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkoxy group substitution. 12. The compound represented by the general formula (I) according to any one of claims 1 to 6 or 8 to 9 or 11 or its stereoisomers, tautomers, mesomers, racemates The rotamers, enantiomers, diastereoisomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, which are of general formula (IIID), general formula (IIIE), general formula (IIIF) or general formula Compounds represented by (IIID-1), general formula (IIIE-1), general formula (IIIF-1) or their stereoisomers, tautomers, mesoforms, racemates, enantiomers isomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, in, X ₁ and X ₂ are each independently selected from -CH ₂ -, -O-, -NH-; n ₁ is 0 or 1; _n2 is 0, 1 or 2; Each R ^c is independently selected from halogen, amino, nitro, cyano, oxo, hydroxyl, mercapto, carboxyl, ester, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclyl, C ₆ -C ₁₀ aryl and 5 to 10 membered heteroaryl; q is 0, 1 or 2, preferably 0 or 1, more preferably 0; Each R ³ is independently selected from hydrogen, halogen, amino, nitro, cyano, oxo, hydroxyl, mercapto, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclic group, C ₆ -C ₁₀ aryl, 5 to 10 membered heteroaryl, the C ₁ - C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclyl, C ₆ - C ₁₀ aryl, 5 to 10 membered heteroaryl is optionally selected from halogen, amino, nitro, cyano, hydroxyl, mercapto, carboxyl, ester, oxo, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6-membered heterocyclic group, C ₆ -C ₁₀ aryl group, 5 to 10-membered heteroaryl group substituted by one or more groups; preferably R ³ is hydrogen; r is 0, 1, 2, 3 or 4, preferably 0, 1 or 2, more preferably 0 or 1; Each R ⁴ is independently selected from halogen, cyano, hydroxyl, mercapto, C ₁ -C ₆ alkyl, -NR ^a R ^b , -OR ^a , said C ₁ -C ₆ alkyl being optionally substituted by halogen ; Preferably ^R is selected from halogen; R ^a and R ^b are each independently selected from hydrogen and C ₁ -C ₆ alkyl; or, R ^a and R ^b form a 5-6 membered nitrogen-containing heterocyclic group together with their connected nitrogen atoms, and the 5-6 membered nitrogen-containing heterocyclic group is optionally selected from halogen, amino, nitro, cyano, oxygen One or more of substituent, hydroxyl, mercapto, carboxyl, ester, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkoxy group substitution; s is 0, 1 or 2; Z, ^R2 and n are as defined in claim 1. 13. according to the compound represented by the general formula (I) described in any one in claim 1 to 5 or 7 to 8 or 10-11 or its stereoisomer, tautomer, mesomer, Racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, which are of the general formula (IVC), general formula (IVD), general formula (IVC-1 ) or the compound represented by general formula (IVD-1) or its stereoisomers, tautomers, mesomers, racemates, enantiomers, diastereoisomers, or its mixture form, or its pharmaceutically acceptable salt, in, Ring E ₁ is C ₃ -C ₆ cycloalkyl or 3 to 6 membered heterocyclic group; Ring E ₂ is C ₃ -C ₆ cycloalkyl or 3 to 6 membered heterocyclic group; Each R ^e is independently selected from halogen, amino, nitro, cyano, oxo, hydroxyl, mercapto, carboxyl, ester, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclyl, C ₆ -C ₁₀ aryl and 5 to 10 membered heteroaryl; t is 0, 1 or 2, preferably 0 or 1, more preferably 0; Each R ³ is independently selected from hydrogen, halogen, amino, nitro, cyano, oxo, hydroxyl, mercapto, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclic group, C ₆ -C ₁₀ aryl, 5 to 10 membered heteroaryl, the C ₁ - C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6 membered heterocyclyl, C ₆ - C ₁₀ aryl, 5 to 10 membered heteroaryl is optionally selected from halogen, amino, nitro, cyano, hydroxyl, mercapto, carboxyl, ester, oxo, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 6-membered heterocyclic group, C ₆ -C ₁₀ aryl group, 5 to 10-membered heteroaryl group substituted by one or more groups; preferably R ³ is hydrogen; r is 0, 1, 2, 3 or 4, preferably 0, 1 or 2, more preferably 0 or 1; Each R ⁴ is independently selected from halogen, cyano, hydroxyl, mercapto, C ₁ -C ₆ alkyl, -NR ^a R ^b , -OR ^a , said C ₁ -C ₆ alkyl being optionally substituted by halogen ; Preferably ^R is selected from halogen; R ^a and R ^b are each independently selected from hydrogen and C ₁ -C ₆ alkyl; or, R ^a and R ^b form a 5-6 membered nitrogen-containing heterocyclic group together with their connected nitrogen atoms, and the 5-6 membered nitrogen-containing heterocyclic group is optionally selected from halogen, amino, nitro, cyano, oxygen One or more of substituent, hydroxyl, mercapto, carboxyl, ester, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkoxy group substitution; s is 0, 1 or 2; Z, ^R2 and n are as defined in claim 1. 14. The compound represented by the general formula (I) according to any one of claims 1 to 13 or its stereoisomers, tautomers, mesomers, racemates, enantiomers isomers, diastereoisomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, wherein, Z is CH. 15. The compound represented by the general formula (I) according to any one of claims 1 to 14 or its stereoisomers, tautomers, mesomers, racemates, enantiomers isomer, diastereoisomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, wherein each ^R is independently selected from hydrogen, halogen, amino, nitro, cyano, oxo, hydroxyl, Mercapto, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, the C ₁ -C ₆ alkyl, C ₁ -C ₆ Alkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl are optionally selected from halogen, amino, nitro, cyano, hydroxyl, mercapto, carboxyl, ester, oxo, C ₁ - C ₆ haloalkyl, C ₁ -C ₆ haloalkoxy is substituted by one or more groups; preferably each R ² is independently selected from hydrogen, hydroxyl and C ₁ -C ₆ alkyl; n is 0, 1, 2, 3, preferably 0 or 1, more preferably 0. 16. The compound represented by the general formula (I) according to any one of claims 1 to 15 or its stereoisomers, tautomers, mesomers, racemates, enantiomers isomers, diastereoisomers, or a mixture thereof, or a pharmaceutically acceptable salt thereof, selected from the group consisting of: 17. A compound represented by general formula (IIIA) or general formula (IIIB) or general formula (IIIC) or its stereoisomer, tautomer, mesomer, racemate, enantiomer A method for preparing isomers, diastereoisomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, comprising the following steps: The compound of the formula IIIAa and the compound of the formula IIIAc obtain the compound of the formula IIIAb in the presence of a catalyst under alkaline conditions; compound; The compound of formula IIIAa and the compound of formula IIIBc obtain the compound of formula IIIBb in the presence of catalyst coupling reaction under basic conditions; compound; The compound of formula IIIAa and the compound of formula IIICc obtain the compound of formula IIICb in the presence of catalyst coupling reaction under basic conditions; compound; in: The catalyst is preferably benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate; The alkaline condition is preferably N,N-diisopropylethylamine; The acidic condition is preferably hydrochloric acid; R ^x is an amino protecting group, preferably Boc; G, Z, R ² , R ³ , X ₁ , X ₂ , R ^c , n, n ₁ , n ₂ , q, r are as defined in claim 9 . 18. A compound represented by general formula (IVA) or general formula (IVB) or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer A method for preparing enantiomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, comprising the following steps: The compound of the formula IIIAa and the compound of the formula IVAc obtain the compound of the formula IVAb in the presence of the catalyst in the presence of a catalyst under alkaline conditions; the compound of the formula IVAb removes the protecting group under acidic conditions to obtain compound; The compound of formula IIIAa and the compound of formula IVBc obtain the compound of formula IVBb in the presence of catalyst coupling reaction under basic conditions; compound; in: The catalyst is preferably benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate; The alkaline condition is preferably N,N-diisopropylethylamine; The acidic condition is preferably hydrochloric acid; R ^x is an amino protecting group, preferably Boc; Ring E ₁ , ring E ₂ , G, Z, R ² , R ³ , ^Re , n, r, t are as defined in claim 10. 19. A pharmaceutical composition containing the compound represented by the general formula (I) according to any one of claims 1 to 16 or its stereoisomers, tautomers, internal Racemate, racemate, enantiomer, diastereoisomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers, diluents or excipient. 20. The compound represented by the general formula (I) according to any one of claims 1 to 16 or its stereoisomers, tautomers, mesomers, racemates, enantiomers isomers, diastereoisomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, or the pharmaceutical composition according to claim 19 in the preparation of medicines for inhibiting AKT1/2/3. 21. The compound represented by the general formula (I) according to any one of claims 1 to 16 or its stereoisomers, tautomers, mesomers, racemates, enantiomers isomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 19 in the preparation for treatment and/or prevention related to AKT1/2/3 kinase activity The purposes of the medicine of the disease, described disease is preferably cancer, especially the cancer that is characterized by the amplification or overexpression of AKT1/2/3, and described cancer is preferably ovarian cancer, breast cancer, prostate cancer, neuroglia tumor, glioma, gastric cancer, fallopian tube cancer, lung cancer, peritoneal tumor, melanoma, brain cancer, esophageal cancer, liver cancer, pancreatic cancer, colorectal cancer, lung cancer, kidney cancer, cervical cancer, skin cancer, neuroblastoma , sarcoma, bone cancer, uterine cancer, endometrial cancer, head and neck cancer, multiple myeloma, lymphoma, non-Hodgkin's lymphoma, non-small cell lung cancer, polycythemia vera, leukemia, thyroid tumors, bladder cancer and Gallbladder cancer, ovarian cancer, breast cancer and prostate cancer are particularly preferred.