Casein kinase 1.epsilon. inhibitor, pharmaceutical composition and application thereof

Abstract

Disclosed is a novel substituted pyrazolopyrimidine compound that inhibits the activity of casein kinase 1ε (CK1ε) and a stereoisomer or stereoisomer mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, and an application thereof in preparation of drugs for treating diseases, disorders or conditions that benefit from the inhibition of the activity of the casein kinase . The compound of the present invention has an inhibitory activity on CK1ε kinase, OCI-LY10 cells and Karpas 299 cells, shows good anti-tumor activity in an OCI-LY10 subcutaneous xenogeneic model and shows excellent synergistic anti-tumor activity in combination with BTK inhibitors. The compound of the present invention has good pharmacokinetics properties and may be used alone or in combination with other drugs to treat diseases, disorders or conditions that benefit from the inhibition of the activity of the casein kinase 1ε , the diseases, disorders or conditions comprising tumors, autoimmune diseases, etc. (see formula I)

Description

CASEIN KINASE 1E INHIBITOR, PHARMACEUTICAL COMPOSITION AND APPLICATION THEREOF FIELD OF TECHNOLOGY

[0001] The present invention belongs to the field of medicines, more specifically relating to a substituted pyrazolopyrimidine casein kinase 1ε inhibitor, a pharmaceutical composition and an application thereof. BACKGROUND

[0002] Protein kinases play an important role in signal transduction, mediating intracellular signal transduction by phosphorylation of substrate proteins, thus being an important type of drug targets. Casein kinase 1 (CK1) belongs to the serine-threonine kinase family with seven isoforms in mammals: <com:Math><semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>γ<annotation encoding="application / x-tex">\gamma< / annotation>< / semantics>< / com:Math>1, <com:Math><semantics>γ<annotation encoding="application / x-tex">\gamma< / annotation>< / semantics>< / com:Math>2, <com:Math><semantics>γ<annotation encoding="application / x-tex">\gamma< / annotation>< / semantics>< / com:Math>3, <com:Math><semantics>δ<annotation encoding="application / x-tex">\delta< / annotation>< / semantics>< / com:Math> and <com:Math><semantics>ϵ<annotation encoding="application / x-tex">\epsilon< / annotation>< / semantics>< / com:Math>. These isoforms are involved in Wnt signal transduction, circadian rhythms, cell signal transduction, membrane transport, DNA replication, DNA damage and RNA metabolism and can regulate organismal functions.

[0003] Phosphorylated substrates of CK1 are involved in a variety of intracellular regulations. For example, the tumor suppressor p53 and oncogene MDM2 are both important proteins that control abnormal cell growth, and substrates for CK1 as well. The isoforms of CK1 kinase (primarily CK1<com:Math><semantics>ϵ<annotation encoding="application / x-tex">\epsilon< / annotation>< / semantics>< / com:Math> and CK1<com:Math><semantics>δ<annotation encoding="application / x-tex">\delta< / annotation>< / semantics>< / com:Math>) have a significant correlation with p53 phosphorylation, p53 stabilization and activation, p53-MDM2 interaction, etc. They are also associated with the formation of centrosome and spindle apparatus during cell division followed by regulations of protein synthesis, or with apoptosis of TRAIL (TNF-related apoptosis-inducing ligand) and FAS.

[0004] Casein kinase 1ε (CK1ε), an important isoform of the CK1 family, is a key regulator of various cell growth and survival processes, which not only plays an important role in the regulation of circadian rhythms, but is equally important in the development and progression of cancer. For example, pharmacological inhibition of CK1s or shRNA-mediated excision of CK1s can hinder the growth and survival of a variety of cancers, including pancreatic cancer, sarcoma, breast cancer, rectal cancer, ovarian caner, leukemia, etc. Studies have demonstrated that the mechanism underlying CK1ε-regulated tumor growth and survival is yet unclear due to the lack of substrate specificity of the CK1 gene, which may be associated with Akt, MYC, and β-catenin (Varghese et al., Scientific Reports, 2018, 8: 13621).

[0005] The Wnt signaling pathway regulates the cell proliferation process, and incidence of mutations in this pathway can lead to various cancers, including skin cancer, liver cancer, brain tumor, and colon cancer. CK1 can promote the degradation of <com:Math><semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>< / com:Math>-catenin by phosphorylation thereof. CK1ε also regulates the Wnt signaling pathway by phosphorylation of Tcf3 as the phosphorylated Tcf3 binds to <com:Math><semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>< / com:Math>-catenin to promote the degradation thereof (Knippschild et al., Cellular Signalling, 2005, 17: 675–689).

[0006] Structures of several types of CK1 kinase inhibitors have been reported, such as 1) PF-4800567, a substituted pyrazolopyrimidine, which is a selective CK1<com:Math><semantics>ϵ<annotation encoding="application / x-tex">\epsilon< / annotation>< / semantics>< / com:Math> inhibitor (IC<com:Sub>50< / com:Sub> = 32nM; Walton et al., J Pharmacol Exp Ther. 2009, 330(2):430-9); 2) PF-670462, a substituted imidazol-5-yl pyrimidine compound, which is a selective CKIε / CKIδ inhibitor (IC<com:Sub>50< / com:Sub> of 7.7nM and 14nM, respectively; Badura et al., J Pharmacol Exp Ther. 2007, 322(2):730-8); 3) CK1- IN-1, an imidazol-5-yl pyridine compound, which is a multitarget inhibitor with IC<com:Sub>50< / com:Sub> of 15nM, 16nM and 73nM for CKIδ, CKIε and p38σ MAPK, respectively (WO2015119579A1). [Image disponible dans le document PDF, Image available in the PDF document]

[0007] In addition, CK1ε, as an important component for the regulation of Wnt signaling pathway, can drive the development and progression of chronic lymphocytic leukemia (CLL). CK1E is significantly upregulated in CLL patients. The development of CLL can be blocked through inhibition of CK1s without affecting the BCR-related pathways; in addition, CK1s can enhance the in-vitro and in-vivo anti-tumor activity of ibrutinib, a BTK inhibitor of the BCR pathway. Therefore, CK1 is also an important anti-tumor target (Janovska et al., Blood, 2019, 11: 1206~1218).

[0008] Therefore, the development of drugs with the capability to inhibit CK1 <com:Math><semantics>ϵ<annotation encoding="application / x-tex">\epsilon< / annotation>< / semantics>< / com:Math> will play an important role in treating a variety of diseases such as cancer and autoimmune diseases. SUMMARY

[0009] It is an object of the present invention to provide a novel substituted pyrazolopyrimidine compound that inhibits the activity of casein kinase <com:Math><semantics>1ε<annotation encoding="application / x-tex">1\varepsilon< / annotation>< / semantics>< / com:Math> (CK1<com:Math><semantics>ε<annotation encoding="application / x-tex">\varepsilon< / annotation>< / semantics>< / com:Math>), and a stereoisomer or stereoisomer mixture thereof, or a pharmaceutically acceptable salt or solvate thereof.

[0010] According to the present invention, there is further provided a pharmaceutical composition comprising the above compound and the stereoisomer or stereoisomer mixture thereof, or the pharmaceutically acceptable salt or solvate thereof. The pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

[0011] According to the present invention, there is also provided an application of the above compound and the stereoisomer or stereoisomer mixture thereof or the pharmaceutically acceptable salt or solvate thereof in preparation of medicines for treating diseases, disorders or conditions that benefit from the inhibition of the activity of the case in kinase <com:Math><semantics>1ε<annotation encoding="application / x-tex">1\varepsilon< / annotation>< / semantics>< / com:Math> (CK1<com:Math><semantics>ε<annotation encoding="application / x-tex">\varepsilon< / annotation>< / semantics>< / com:Math>).

[0012] The following technical scheme is adopted in the present invention:

[0013] The CK1 inhibitor provided in the present invention has a structure of general formula I: [Image disponible dans le document PDF, Image available in the PDF document]

[0014] or a stereoisomer thereof, or a stereoisomer mixture thereof, or a pharmaceutically acceptable salt or solvate thereof;

[0015] wherein:

[0016] <com:Math><semantics>R1<annotation encoding="application / x-tex">R_1< / annotation>< / semantics>< / com:Math> is H, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> alkyl, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> haloalkyl, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> alkoxy, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> haloalkoxy, substituted or unsubstituted C<com:Sub>2< / com:Sub>-C<com:Sub>10< / com:Sub> alkenyl, substituted or unsubstituted C<com:Sub>2< / com:Sub>-C<com:Sub>10< / com:Sub> alkinyl, halogen, cyano, amino, nitro or hydroxy;

[0017] m is an integer between 1 and 4; and when m is greater than 1, the plurality of R<com:Sub>1< / com:Sub> may be the same or different, independently of each other;

[0018] <com:Math><semantics>R2<annotation encoding="application / x-tex">R_2< / annotation>< / semantics>< / com:Math> is H, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> alkyl, substituted or unsubstituted <com:Math><semantics>C3<annotation encoding="application / x-tex">C_3< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C10<annotation encoding="application / x-tex">C_{10}< / annotation>< / semantics>< / com:Math> cycloalkyl, or substituted or unsubstituted C<com:Sub>5< / com:Sub>-C<com:Sub>12< / com:Sub> aryl;

[0019] R<com:Sub>3< / com:Sub> is H, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> alkyl, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> haloalkyl, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> alkoxy, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> haloalkoxy, halogen, cyano, amino, nitro or hydroxy;

[0020] X is CR<com:Sub>4< / com:Sub> or N;

[0021] R<com:Sub>4< / com:Sub> is H, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> alkyl, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> haloalkyl, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> alkoxy, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> haloalkoxy, halogen, cyano, amino or hydroxy;

[0022] B is selected from substituted or unsubstituted C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkyl, substituted or unsubstituted C<com:Sub>3< / com:Sub>-C<com:Sub>10< / com:Sub> cycloalkyl, substituted or unsubstituted 3- to 10-membered heterocycloalkyl, substituted or unsubstituted C<com:Sub>5< / com:Sub>-C<com:Sub>12< / com:Sub> aryl, and substituted or unsubstituted 5- to 12-membered heteroaryl.

[0023] As a preferred scheme, the R<com:Sub>3< / com:Sub> and R<com:Sub>4< / com:Sub> are not H simultaneously.

[0024] Further, a preferable compound of the present invention has a structure of general formula II-A, II-B or II-C: [Image disponible dans le document PDF, Image available in the PDF document]

[0025] or a stereoisomer thereof, or a stereoisomer mixture thereof, or a pharmaceutically acceptable salt or solvate thereof;

[0026] wherein: the definitions of <com:Math><semantics>R1<annotation encoding="application / x-tex">R_1< / annotation>< / semantics>< / com:Math>, m, <com:Math><semantics>R2<annotation encoding="application / x-tex">R_2< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>R3<annotation encoding="application / x-tex">R_3< / annotation>< / semantics>< / com:Math>, X, <com:Math><semantics>R4<annotation encoding="application / x-tex">R_4< / annotation>< / semantics>< / com:Math> and B are the same as defined in general formula I;

[0027] R<com:Sub>5< / com:Sub> is H, substituted or unsubstituted C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkyl, substituted or unsubstituted C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> haloalkyl, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> alkoxy, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> haloalkoxy, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> hydroxyalkyl, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>- C<com:Sub>6< / com:Sub> halohydroxyalkyl, halogen, cyano, amino, nitro or hydroxy;

[0028] o is an integer between 1 and 5;

[0029] R<com:Sub>6< / com:Sub> and R<com:Sub>6< / com:Sub>' are independently selected from H, substituted or unsubstituted C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkyl, substituted or unsubstituted C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> haloalkyl, substituted or unsubstituted C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkoxy, substituted or unsubstituted C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> haloalkoxy, halogen, cyano, amino or hydroxy, or, R<com:Sub>6< / com:Sub> and R<com:Sub>6< / com:Sub>' are joined to form a 3- to 6-membered cycloalkyl or heterocycloalkyl;

[0030] n is absent, or is 0, 1 or 2; wherein:

[0031] when n is absent, i.e., when Y and Z are not joined to form a ring, Y is CR<com:Sub>7< / com:Sub>R<com:Sub>7< / com:Sub>' or NR<com:Sub>8< / com:Sub>; and Z is CHR<com:Sub>7< / com:Sub>R<com:Sub>7< / com:Sub>', OH or NHR<com:Sub>8< / com:Sub>; R<com:Sub>7< / com:Sub> in Y and Z may be the same or different, independently of each other; similarly, R<com:Sub>7< / com:Sub>' in Y and Z may be the same or different, independently of each other; and

[0032] when n is 0, 1 or 2, Y is CH or N; and Z is <com:Math><semantics>CR7R7<annotation encoding="application / x-tex">CR_7R_7< / annotation>< / semantics>< / com:Math>, O or <com:Math><semantics>NR8<annotation encoding="application / x-tex">NR_8< / annotation>< / semantics>< / com:Math>;

[0033] <com:Math><semantics>R7<annotation encoding="application / x-tex">R_7< / annotation>< / semantics>< / com:Math> and <com:Math><semantics>R7<annotation encoding="application / x-tex">R_7< / annotation>< / semantics>< / com:Math>' are independently selected from H, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> alkyl, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> haloalkyl, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> alkoxy, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> haloalkoxy, substituted or unsubstituted <com:Math><semantics>C3<annotation encoding="application / x-tex">C_3< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C10<annotation encoding="application / x-tex">C_{10}< / annotation>< / semantics>< / com:Math> cycloalkyl, substituted or unsubstituted 3- to 10-membered heterocycloalkyl, halogen, cyano, amino or hydroxy; or, R<com:Sub>7< / com:Sub> and R<com:Sub>7< / com:Sub>' are joined to form a 3- to 6-membered cycloalkyl or heterocycloalkyl;

[0034] <com:Math><semantics>R8<annotation encoding="application / x-tex">R_8< / annotation>< / semantics>< / com:Math> is H, alkyl (preferably, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> alkyl or substituted or unsubstituted C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> haloalkyl), substituted or unsubstituted C<com:Sub>3< / com:Sub>-C<com:Sub>10< / com:Sub> cycloalkyl, substituted or unsubstituted 3- to 10-membered heterocycloalkyl, -C(O)-R<com:Sub>9< / com:Sub> or -S(O)<com:Sub>2< / com:Sub>-R<com:Sub>10< / com:Sub>;

[0035] R<com:Sub>9< / com:Sub> is substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> alkyl, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> cycloalkyl; and

[0036] <com:Math><semantics>R10<annotation encoding="application / x-tex">R_{10}< / annotation>< / semantics>< / com:Math> is substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> alkyl, or substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> cycloalkyl.

[0037] Even further, a preferable compound of the present invention has a structure of general formula III-A, III-B or III-C: [Image disponible dans le document PDF, Image available in the PDF document]

[0038] or a stereoisomer thereof, or a stereoisomer mixture thereof, or a pharmaceutically acceptable salt or solvate thereof;

[0039] wherein, the definitions of R<com:Sub>1< / com:Sub>, R<com:Sub>2< / com:Sub>, R<com:Sub>3< / com:Sub>, R<com:Sub>4< / com:Sub>, R<com:Sub>5< / com:Sub>, R<com:Sub>6< / com:Sub>, R<com:Sub>6< / com:Sub>', Y, Z and n are the same as defined in general formulas II-A, II-B and II-C.

[0040] Even further, a preferable compound of the present invention has a structure of general formula IV-A, IV-B or IV-C: [Image disponible dans le document PDF, Image available in the PDF document]

[0041] or a stereoisomer thereof, or a stereoisomer mixture thereof, or a pharmaceutically acceptable salt or solvate thereof;

[0042] wherein:

[0043] <com:Math><semantics>R1<annotation encoding="application / x-tex">R_1< / annotation>< / semantics>< / com:Math> is selected from H, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> alkyl, and halogen;

[0044] m is selected from integers of 1 to 4;

[0045] R<com:Sub>2< / com:Sub> is selected from methyl, ethyl, isopropyl and cyclopropyl;

[0046] R<com:Sub>5< / com:Sub> is selected from H, substituted or unsubstituted C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkyl, and halogen;

[0047] o is selected from integers of 1 to 4;

[0048] R<com:Sub>6< / com:Sub> and R<com:Sub>6< / com:Sub>' are selected from H, substituted or unsubstituted C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkyl, and halogen;

[0049] Y is selected from N or CH;

[0050] Z is selected from CH<com:Sub>2< / com:Sub>, O and NR<com:Sub>8< / com:Sub>;

[0051] R<com:Sub>8< / com:Sub> is selected from H, substituted or unsubstituted C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkyl, and -C(O)-R<com:Sub>9< / com:Sub>; and

[0052] <com:Math><semantics>R9<annotation encoding="application / x-tex">R_9< / annotation>< / semantics>< / com:Math> is selected from substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> alkyl.

[0053] Even further, a preferable compound of the present invention has a structure of general formula V-A, V-B or V-C: [Image disponible dans le document PDF, Image available in the PDF document]

[0054] or a stereoisomer thereof, or a stereoisomer mixture thereof, or a pharmaceutically acceptable salt or solvate thereof;

[0055] wherein: the definitions of <com:Math><semantics>R1<annotation encoding="application / x-tex">R_1< / annotation>< / semantics>< / com:Math>, m, <com:Math><semantics>R5<annotation encoding="application / x-tex">R_5< / annotation>< / semantics>< / com:Math> and Z are the same as defined in general formulas IV-A, IV-B and IV-C; and R<com:Sub>2< / com:Sub> is methyl, ethyl or isopropyl.

[0056] Preferably, a preferable compound of the present invention has a structure of general formula V-A, V-B or V-C, wherein:

[0057] R<com:Sub>1< / com:Sub> is selected from H and halogen (preferably Cl and F);

[0058] R<com:Sub>2< / com:Sub> is selected from methyl, ethyl, isopropyl, cyclopropyl and phenyl;

[0059] m is 1, 2 or 3;

[0060] R<com:Sub>3< / com:Sub> is selected from F, Cl, CN and H; and

[0061] X is C, N or CH.

[0062] As a preferable scheme, B is phenyl, pyridinyl (further preferably at 3-position, i.e., pyridin-3-yl), halogen substituted pyridinyl (further preferably Cl or F substituted pyridinyl; preferably at 3-position, i.e., Cl or F substituted pyridin-3-yl), cyclohexyl, piperidinyl (further preferably at 4-position (i.e., piperidin-4-yl) and at 1-position (i.e., piperidin-1-yl)), N- acetylpiperidinyl (further preferably at 4-position, i.e., N-acetylpiperidin-4-yl), N-(2- trifluoroethyl) substituted piperidinyl (further preferably at 4-position, i.e., N-(2- trifluoroethyl) substituted piperidin-4-yl), morpholino (further preferably at N-position or C- position), piperazinyl (further preferably at N-position), dimethyl (further preferably dimethyl substituent at C-position) substituted piperazinyl (further preferably at N-position), trifluoromethyl substituted (further preferably trifluoromethyl substituent at C-position) piperazinyl (further preferably at N-position), trifluoroethyl substituted (further preferably trifluoroethyl substituent at N-position) piperazinyl (further preferably at N-position), methyl substituted (further preferably methyl substituent at N-position) piperazinyl (further preferably at N-position), acetyl substituted (further preferably acetyl substituent at N- position) piperazinyl (further preferably at N-position), 3-methoxypropyl and 2- (tetrahydrofuran-2-yl)ethyl.

[0063] As preferred, the CK1ɛ inhibitor is preferably the following specific compounds: [Image disponible dans le document PDF, Image available in the PDF document] [Image disponible dans le document PDF, Image available in the PDF document] [Image disponible dans le document PDF, Image available in the PDF document]

[0064] and stereoisomers of the above formulas, or stereoisomer mixtures thereof, or pharmaceutically acceptable salts or solvates thereof.

[0065] As preferred, the CK1<com:Math><semantics>ϵ<annotation encoding="application / x-tex">\epsilon< / annotation>< / semantics>< / com:Math> inhibitor is preferably one of the following compounds: [Image disponible dans le document PDF, Image available in the PDF document]

[0066] As preferred, the compound is the following compounds:

[0067] 2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1- yl)ethyl)-3-phenylquinazolin-4(3H)-one (II-A-1)

[0068] (S)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4- d]pyrimidin-1-yl)ethyl)-3-phenylquinazolin-4(3H)-one (II-A-1a)

[0069] (R)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4- d]pyrimidin-1-yl)ethyl)-3-phenylquinazolin-4(3H)-one (II-A-1b)

[0070] 2-(1-(4-amino-3-(2,3-difluoro-4-isopropoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin- 1-yl)ethyl)-3-phenylquinazolin-4(3H)-one (II-A-2)

[0071] (S)-2-(1-(4-amino-3-(2,3-difluoro-4-isopropoxyphenyl)-1H-pyrazolo[3,4- d]pyrimidin-1-yl)ethyl)-3-phenylquinazolin-4(3H)-one (II-A-2a)

[0072] (R)-2-(1-(4-amino-3-(2,3-difluoro-4-isopropoxyphenyl)-1H-pyrazolo[3,4- d]pyrimidin-1-yl)ethyl)-3-phenylquinazolin-4(3H)-one (II-A-2b)

[0073] 2-(1-(4-amino-3-(4-ethoxy-2,3-difluorophenyl)-1H-pyrazolo[3,4-d]pyrimidin-1- yl)ethyl)-3-phenylquinazolin-4(3H)-one (II-A-3)

[0074] 2-(1-(4-amino-3-(4-cyclopropoxy-2,3-difluorophenyl)-1H-pyrazolo[3,4- d]pyrimidin-1-yl)ethyl)-3-phenylquinazolin-4(3H)-one (II-A-4)

[0075] 2-(1-(4-amino-3-(3-chloro-2-fluoro-4-methoxyphenyl)-1H-pyrazolo[3,4- d]pyrimidin-1-yl)ethyl)-3-phenylquinazolin-4(3H)-one (II-A-5)

[0076] 3-(4-amino-1-(1-(4-oxo-3-phenyl-3,4-dihydroquinazolin-2-yl)ethyl)-1H- pyrazolo[3,4-d]pyrimidin-3-yl)-2-fluoro-6-methoxybenzonitrile (II-A-6)

[0077] 2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1- yl)ethyl)-5-chloro-3-phenylquinazolin-4(3H)-one (H-A-7)

[0078] (S)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4- d]pyrimidin-1-yl)ethyl)-5-chloro-3-phenylquinazolin-4(3H)-one (H-A-7a)

[0079] (R)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4- d]pyrimidin-1-yl)ethyl)-5-chloro-3-phenylquinazolin-4(3H)-one (H-A-7b)

[0080] 2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1- yl)ethyl)-5-fluoro-3-phenylquinazolin-4(3H)-one (II-A-8)

[0081] (S)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4- d]pyrimidin-1-yl)ethyl)-5-fluoro-3-phenylquinazolin-4(3H)-one (H-A-8a)

[0082] (R)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4- d]pyrimidin-1-yl)ethyl)-5-fluoro-3-phenylquinazolin-4(3H)-one (II-A-8b)

[0083] 2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1- yl)ethyl)-6-fluoro-3-phenylquinazolin-4(3H)-one (II-A-9)

[0084] 2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1- yl)ethyl)-5-chloro-8-fluoro-3-phenylquinazolin-4(3H)-one (II-A-10)

[0085] 2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1- yl)ethyl)-5,8-difluoro-3-phenylquinazolin-4(3H)-one (H-A-11)

[0086] 2-(1-(4-amino-3-(2,6-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1- yl)ethyl)-3-phenylquinazolin-4(3H)-one (II-A-12)

[0087] 2-(1-(4-amino-3-(2-fluoro-6-methoxpyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-1- yl)ethyl)-3-phenylquinazolin-4(3H)-one (II-A-13)

[0088] 2-(1-(4-amino-3-(2-fluoro-6-isopropoxypyridin-3-yl)-1H-pyrazolo[3,4- d]pyrimidin-1-yl)ethyl)-3-phenylquinazolin-4(3H)-one (II-A-14)

[0089] 2-(1-(4-amino-3-(2-fluoro-6-pheoxypyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-1- yl)ethyl)-3-phenylquinazolin-4(3H)-one (II-A-15)

[0090] 2-(1-(4-amino-3-(2,3-difluoro-4-isopropoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin- 1-yl)ethyl)-5-fluoro-3-phenylquinazolin-4(3H)-one (II-A-16)

[0091] 2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1- yl)ethyl)-3-(pyridin-3-yl)quinazolin-4(3H)-one (II-B-1)

[0092] 2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1- yl)ethyl)-5-chloro-3-(pyridin-3-yl)quinazolin-4(3H)-one (H-B-2)

[0093] (S)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4- d]pyrimidin-1-yl)ethyl)-5-chloro-3-(pyridin-3-yl)quinazolin-4(3H)-one (H-B-2a)

[0094] (R)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4- d]pyrimidin-1-yl)ethyl)-5-chloro-3-(pyridin-3-yl)quinazolin-4(3H)-one (H-B-2b)

[0095] 2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1- yl)ethyl)-5-chloro-3-(5-fluoropyridin-3-yl)quinazolin-4(3H)-one (H-B-3)

[0096] 2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1- yl)ethyl)-5-chloro-3-cyclohexylquinazolin-4(3H)-one (II-C-1)

[0097] 2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1- yl)ethyl)-5-chloro-3-(piperidin-3-yl)quinazolin-4(3H)-one (II-C-2)

[0098] 3-(1-acetylpiperidin-4-yl)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H- pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloroquinazolin-4(3H)-one (II-C-3)

[0099] 2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1- yl)ethyl)-5-chloro-3-(1-(2,2,2-trifluoroethyl)piperidin-4-yl)quinazolin-4(3H)-one (II-C-4)

[0100] 2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1- yl)ethyl)-5-chloro-3-(piperidin-1-yl)quinazolin-4(3H)-one (H-C-5)

[0101] 2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1- yl)ethyl)-5-chloro-3-morpholinylquinazolin-4(3H)-one (II-C-6)

[0102] 2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1- yl)ethyl)-5-chloro-3-(piperazin-1-yl)quinazolin-4(3H)-one (II-C-7)

[0103] 2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1- yl)ethyl)-5-chloro-3-(3,3-dimethylpiperazin-1-yl)quinazolin-4(3H)-one (H-C-8)

[0104] 2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1- yl)ethyl)-5-chloro-3-(3-(trifluoromethyl)piperazin-1-yl)quinazolin-4(3H)-one (II-C-9)

[0105] 2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1- yl)ethyl)-5-chloro-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)quinazolin-4(3H)-one (II-C-10)

[0106] 2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1- yl)ethyl)-5-chloro-3-(4-methylpiperazin-1-yl)quinazolin-4(3H)-one (II-C-11) [Image disponible dans le document PDF, Image available in the PDF document] pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloroquinazolin-4(3H)-one (H-C-12)

[0108] 2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1- yl)ethyl)-3-(3-methoxypropyl)quinazolin-4(3H)-one (H-C-13)

[0109] 2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1- yl)ethyl)-5-chloro-3-(3-methoxypropyl)quinazolin-4(3H)-one (II-C-14)

[0110] 2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1- yl)ethyl)-5-chloro-3-(2-(tetrahydrofuran-2-yl)ethyl)quinazolin-4(3H)-one (II-C-15)

[0111] or a stereoisomer thereof, or a stereoisomer mixture thereof, or a pharmaceutically acceptable salt or solvate thereof.

[0112] As preferred, the compound is the following compounds:

[0113] (S)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4- d]pyrimidin-1-yl)ethyl)-3-phenylquinazolin-4(3H)-one hydrochloride (H-D-1)

[0114] (S)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4- d]pyrimidin-1-yl)ethyl)-3-phenylquinazolin-4(3H)-one sulfate (II-D-2)

[0115] (S)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4- d]pyrimidin-1-yl)ethyl)-3-phenylquinazolin-4(3H)-one p-toluenesulfonate (H-D-3)

[0116] (S)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4- d]pyrimidin-1-yl)ethyl)-3-phenylquinazolin-4(3H)-one methanesulfonate (II-D-4)

[0117] (S)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4- d]pyrimidin-1-yl)ethyl)-5-chloro-3-phenylquinazolin-4(3H)-one hydrochloride (II-D-5)

[0118] (S)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4- d]pyrimidin-1-yl)ethyl)-5-chloro-3-phenylquinazolin-4(3H)-one sulfate (H-D-6)

[0119] (S)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4- d]pyrimidin-1-yl)ethyl)-5-chloro-3-phenylquinazolin-4(3H)-one phosphate (II-D-7)

[0120] (S)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4- d]pyrimidin-1-yl)ethyl)-5-chloro-3-phenylquinazolin-4(3H)-one p-toluenesulfonate (H-D-8)

[0121] (S)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4- d]pyrimidin-1-yl)ethyl)-5-chloro-3-phenylquinazolin-4(3H)-one methanesulfonate (H-D-9)

[0122] (S)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4- d]pyrimidin-1-yl)ethyl)-5-chloro-3-phenylquinazolin-4(3H)-one citrate (H-D-10)

[0123] (S)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4- [Image disponible dans le document PDF, Image available in the PDF document] [Image disponible dans le document PDF, Image available in the PDF document] [Image disponible dans le document PDF, Image available in the PDF document] [Image disponible dans le document PDF, Image available in the PDF document] [Image disponible dans le document PDF, Image available in the PDF document] [Image disponible dans le document PDF, Image available in the PDF document] [Image disponible dans le document PDF, Image available in the PDF document]

[0127] (S)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4- d]pyrimidin-1-yl)ethyl)-5-chloro-3-phenylquinazolin-4(3H)-one benzoate (H-D-15)

[0128] (S)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4- d]pyrimidin-1-yl)ethyl)-5-chloro-3-phenylquinazolin-4(3H)-one maleate (H-D-16)

[0129] (S)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4- d]pyrimidin-1-yl)ethyl)-5-chloro-3-phenylquinazolin-4(3H)-one oxalate (II-D-17)

[0130] or a stereoisomer thereof, or a stereoisomer mixture or solvate thereof.

[0131] There is further provided in the present invention a preparation method for the compound of general formula I, the method comprising the following 3 methods:

[0132] 1) Method 1: alkylation of 3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (compound 5) by compound 4 in the presence of a base yields compound 10, which then undergoes coupling reaction with compound 7 to give compound of general formula I: [Image disponible dans le document PDF, Image available in the PDF document]

[0133] In the reaction of the first step, the reaction solvent can be selected from THF, MeCN, DMSO, etc. in addition to DMF. The mole ratio of compound 4 to compound 5 is about <com:Math><semantics>1:(1∼3)<annotation encoding="application / x-tex">1:(1\sim3)< / annotation>< / semantics>< / com:Math>, further preferably <com:Math><semantics>1:(1−2)<annotation encoding="application / x-tex">1:(1-2)< / annotation>< / semantics>< / com:Math>. The base may be selected from potassium carbonate, caesium carbonate, sodium carbonate, etc.; the mole ratio of the base to compound 5 is about <com:Math><semantics>(1∼5):1<annotation encoding="application / x-tex">(1\sim5):1< / annotation>< / semantics>< / com:Math>, further preferably <com:Math><semantics>(1.5∼2.5):1<annotation encoding="application / x-tex">(1.5\sim2.5):1< / annotation>< / semantics>< / com:Math>. The reaction temperature is between room temperature and 60°C.

[0134] In the reaction of the second step, compound 10 reacts with compound 7 in the presence of a base (sodium carbonate or potassium carbonate) and a catalyst (such as a palladium catalyst). The catalyst is preferably tetrakis(triphenylphosphine)palladium, etc. The mole ratio of compound 10 to compound 7 is <com:Math><semantics>1:(1∼3)<annotation encoding="application / x-tex">1:(1\sim3)< / annotation>< / semantics>< / com:Math>, further preferably <com:Math><semantics>1:(1∼2)<annotation encoding="application / x-tex">1:(1\sim2)< / annotation>< / semantics>< / com:Math>. The mole ratio of the base to compound 10 is <com:Math><semantics>1:(1∼4)<annotation encoding="application / x-tex">1:(1\sim4)< / annotation>< / semantics>< / com:Math>, further preferably <com:Math><semantics>1:(2∼3)<annotation encoding="application / x-tex">1:(2\sim3)< / annotation>< / semantics>< / com:Math>. The amount of the catalyst added is 0.05~1.0% of the moles of compound 10. The reaction solvent is a mixture of an organic solvent, such as dioxane, tetrahydrofuran and N,N-dimethylformamide, with water, etc., in a volume ratio of <com:Math><semantics>(2∼10)<annotation encoding="application / x-tex">(2\sim10)< / annotation>< / semantics>< / com:Math>: 1, further preferably <com:Math><semantics>(2∼5)<annotation encoding="application / x-tex">(2\sim5)< / annotation>< / semantics>< / com:Math>:1. The reaction temperature is 80~120°C.

[0135] When using this method, as preferred, the <com:Math><semantics>R2<annotation encoding="application / x-tex">R_2< / annotation>< / semantics>< / com:Math> is preferably <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C5<annotation encoding="application / x-tex">C_5< / annotation>< / semantics>< / com:Math> alkyl group (preferably methyl, ethyl, isopropyl, propyl, cyclopropyl) or phenyl, the R<com:Sub>3< / com:Sub> is preferably H, halogen (preferably F), and X is C-halogen (preferably C-Cl, C-F, etc.), C-CN, CH or N.

[0136] 2) Method 2: 3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (compound 5) reacts with triphenylchloromethane in the presence of a base to produce compound 6, which then reacts with compound 7 by coupling to produce compound 8, which alkylates with compound 4 in the presence of a base after deprotection to produce a compound of general formula I: [Image disponible dans le document PDF, Image available in the PDF document]

[0137] In the first step of this method, the mole ratio of triphenylchloromethane to compound 5 is <com:Math><semantics>(1∼5):1<annotation encoding="application / x-tex">(1\sim5):1< / annotation>< / semantics>< / com:Math>, preferably <com:Math><semantics>(1.5∼2.5):1<annotation encoding="application / x-tex">(1.5\sim2.5):1< / annotation>< / semantics>< / com:Math>; a base (preferably potassium carbonate or caesium carbonate) can be added in this step in a mole ratio of <com:Math><semantics>(1∼8)<annotation encoding="application / x-tex">(1\sim8)< / annotation>< / semantics>< / com:Math>:1 between the base and compound 5, preferably <com:Math><semantics>(2∼5)<annotation encoding="application / x-tex">(2\sim5)< / annotation>< / semantics>< / com:Math>:1. The reaction temperature is between 60 and <com:Math><semantics>100∘<annotation encoding="application / x-tex">100^{\circ}< / annotation>< / semantics>< / com:Math>C.

[0138] In the second step, compound 6 reacts with compound 7 in the presence of a base (sodium carbonate, potassium carbonate) and a catalyst (such as a palladium catalyst). The catalyst is preferably tetrakis(triphenylphosphine)palladium, etc. The mole ratio of compound 6 to compound 7 is <com:Math><semantics>1:(1∼3)<annotation encoding="application / x-tex">1:(1\sim3)< / annotation>< / semantics>< / com:Math>, further preferably <com:Math><semantics>1:(1∼2)<annotation encoding="application / x-tex">1:(1\sim2)< / annotation>< / semantics>< / com:Math>. The mole ratio of the base to compound 6 is <com:Math><semantics>(1∼4):1<annotation encoding="application / x-tex">(1\sim4):1< / annotation>< / semantics>< / com:Math>, further preferably <com:Math><semantics>(2∼3):1<annotation encoding="application / x-tex">(2\sim3):1< / annotation>< / semantics>< / com:Math>. The amount of catalyst added is <com:Math><semantics>0.05∼1.0%<annotation encoding="application / x-tex">0.05\sim1.0\%< / annotation>< / semantics>< / com:Math> of the moles of compound 6. The reaction solvent is a mixture of an organic solvent, such as dioxane, tetrahydrofuran and N,N-dimethylformamide, with water, etc., in a volume ratio of <com:Math><semantics>(2∼10)<annotation encoding="application / x-tex">(2\sim10)< / annotation>< / semantics>< / com:Math>:1, further preferably <com:Math><semantics>(2∼5)<annotation encoding="application / x-tex">(2\sim5)< / annotation>< / semantics>< / com:Math>:1. The reaction temperature is between 80 and 120°C.

[0139] In the third step, TFA and triisopropylsilane are added during the reaction, and the reaction solvent can be dichloromethane, chloroform, etc. The mole ratio of TFA to compound 8 is <com:Math><semantics>(2∼8):1<annotation encoding="application / x-tex">(2\sim8):1< / annotation>< / semantics>< / com:Math>, further preferably <com:Math><semantics>(3∼6):1<annotation encoding="application / x-tex">(3\sim6):1< / annotation>< / semantics>< / com:Math>. The mole ratio of triisopropylsilane to compound 8 is <com:Math><semantics>(2∼8):1<annotation encoding="application / x-tex">(2\sim8):1< / annotation>< / semantics>< / com:Math>, further preferably <com:Math><semantics>(3∼6):1<annotation encoding="application / x-tex">(3\sim6):1< / annotation>< / semantics>< / com:Math>.

[0140] In the fourth step, the reaction solvent can be selected from THF, MeCN, DMSO, etc. in addition to DMF. The mole ratio of compound 9 to compound 4 is about <com:Math><semantics>1:(1∼3)<annotation encoding="application / x-tex">1:(1\sim3)< / annotation>< / semantics>< / com:Math>, further preferably 1:<com:Math><semantics>(1∼2)<annotation encoding="application / x-tex">(1\sim2)< / annotation>< / semantics>< / com:Math>. A base, selected from potassium carbonate, caesium carbonate, sodium carbonate, etc., can be added during the reaction in a mole ratio of about <com:Math><semantics>(1∼5)<annotation encoding="application / x-tex">(1\sim5)< / annotation>< / semantics>< / com:Math>:1 between the base and compound 9, further preferably <com:Math><semantics>(1.5∼2.5)<annotation encoding="application / x-tex">(1.5\sim2.5)< / annotation>< / semantics>< / com:Math>:1. The reaction temperature is between room temperature and 60°C.

[0141] In this method, as preferred, <com:Math><semantics>R1<annotation encoding="application / x-tex">R_1< / annotation>< / semantics>< / com:Math> is halogen (preferably one or more of Cl, one or more of F, or a combination thereof); <com:Math><semantics>R2<annotation encoding="application / x-tex">R_2< / annotation>< / semantics>< / com:Math> is <com:Math><semantics>C1∼C5<annotation encoding="application / x-tex">C_1 \sim C_5< / annotation>< / semantics>< / com:Math> (methyl, ethyl, isopropyl etc.); and <com:Math><semantics>R3<annotation encoding="application / x-tex">R_3< / annotation>< / semantics>< / com:Math> is H, halogen, etc. B is preferably phenyl, halogen (Cl, F) substituted or unsubstituted pyridinyl, piperidinyl, methyl substituted piperazinyl, Boc substituted piperidinyl, N-heteropiperidinyl, alkyl substituted piperazinyl, trifluoromethyl substituted piperazinyl, trifluoromethyl substituted alkyl substituted piperazinyl, acetyl substituted piperazinyl, substituted propyl, propyl, substituted ethyl, etc.

[0142] 3) Method 3 (synthesis of a chiral compound): [Image disponible dans le document PDF, Image available in the PDF document] [Image disponible dans le document PDF, Image available in the PDF document] - -

[0143] The method first employs chiral methyl D-lactate and methyl L-lactate to produce (R)-12 or (S)-12 by a multi-step reaction, followed by a Mitsunobu reaction with intermediate 9 in the presence of a phosphine ligand and azodicarboxylic acid ester to give (S)-I or (R)-I, respectively.

[0144] The reaction solvent of the Mitsunobu reaction may be THF, dichloromethane, toluene, acetonitrile, DMF, etc., in a volume ratio of (2~10):1, preferably (2~5):1 to intermediate 9; the phosphine ligand may be selected from triphenylphosphine, tri-tert- butylphosphine, in a molar ratio of <com:Math><semantics>(1∼4):1<annotation encoding="application / x-tex">(1\sim4):1< / annotation>< / semantics>< / com:Math>, preferably <com:Math><semantics>(1.5∼3):1<annotation encoding="application / x-tex">(1.5\sim3):1< / annotation>< / semantics>< / com:Math> to intermediate 9; the azodicarboxylic acid ester may be selected from DEAD and DIAD, in a molar ratio of <com:Math><semantics>(1∼4):1<annotation encoding="application / x-tex">(1\sim4):1< / annotation>< / semantics>< / com:Math>, preferably (1.5~3):1 to intermediate 9. The reaction temperature is between 15 and 30°C, preferably room temperature.

[0145] When using this method, as preferred, the R<com:Sub>2< / com:Sub> is preferably C<com:Sub>1< / com:Sub>-C<com:Sub>5< / com:Sub> alkyl (preferably methyl, ethyl, isopropyl, propyl, cyclopropyl) or phenyl, the R<com:Sub>3< / com:Sub> is preferably H, halogen (preferably F), and X is C-halogen (preferably C-Cl, C-F, etc.), C-CN, CH or N.

[0146] It is another object of the present invention to provide an intermediate as shown by the following compound 9 and an application thereof in preparation of drugs for treating diseases, disorders or conditions that benefit from the inhibition of the activity of the casein kinase 1ε: [Image disponible dans le document PDF, Image available in the PDF document]

[0147] wherein:

[0148] <com:Math><semantics>R2<annotation encoding="application / x-tex">R_2< / annotation>< / semantics>< / com:Math> is H, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> alkyl, substituted or unsubstituted <com:Math><semantics>C3<annotation encoding="application / x-tex">C_3< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C10<annotation encoding="application / x-tex">C_{10}< / annotation>< / semantics>< / com:Math> cycloalkyl, or substituted or unsubstituted C<com:Sub>5< / com:Sub>-C<com:Sub>12< / com:Sub> aryl; preferably R<com:Sub>2< / com:Sub> is methyl, ethyl, isopropyl or cyclopropyl;

[0149] R<com:Sub>3< / com:Sub> is H, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> alkyl, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> haloalkyl, substituted or unsubstituted C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkoxy, substituted or unsubstituted C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> haloalkoxy, halogen, cyano, amino, nitro or hydroxy; preferably R<com:Sub>3< / com:Sub> is F, Cl, CN or H;

[0150] X is CR<com:Sub>4< / com:Sub> or N;

[0151] R<com:Sub>4< / com:Sub> is H, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> alkyl, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> haloalkyl, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> alkoxy, substituted or unsubstituted <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> haloalkoxy, halogen, cyano, amino or hydroxy; preferably R<com:Sub>4< / com:Sub> is F;

[0152] As a preferable scheme, the <com:Math><semantics>R3<annotation encoding="application / x-tex">R_3< / annotation>< / semantics>< / com:Math> and <com:Math><semantics>R4<annotation encoding="application / x-tex">R_4< / annotation>< / semantics>< / com:Math> are not H simultaneously.

[0153] Preferably, X is CF, N or CH.

[0154] As preferred, the intermediate as shown by compound 9 is preferably a compound as shown by the following formula: [Image disponible dans le document PDF, Image available in the PDF document]

[0155] where R<com:Sub>2< / com:Sub> is selected from methyl, ethyl, isopropyl and cyclopropyl.

[0156] Description of Terms:

[0157] The term "aryl" as used herein is obtained by removing a H from an aromatic ring, the aromatic ring referring to an all-carbon monocyclic or fused polycyclic group of 5 to 12 carbon atoms with a fully conjugated <com:Math><semantics>π<annotation encoding="application / x-tex">\pi< / annotation>< / semantics>< / com:Math>-electron system. Non-limiting examples of the aromatic ring are: benzene ring, naphthalene ring and anthracene ring. Aromatic rings may be unsubstituted or substituted. A substituent of the aromatic ring is selected from halogen (preferably fluorine, chlorine, bromine or iodine), cyano, nitro, amino, hydroxy, C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkyl (preferably methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, etc.), <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> hydroxyalkyl (preferably hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxyisopropyl, etc.), <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> alkoxy (preferably methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, etc.), C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkyl halide (preferably halomethyl, haloethyl, halopropyl, haloisopropyl, halobutyl, haloisobutyl, sec-butyl halide, tert-butyl halide, etc.), C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> hydroxyalkyl halide (preferably halohydroxymethyl, halohydroxyethyl, halohydroxypropyl, halohydroxyisopropyl, etc.), C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkoxy halide (preferably halomethoxy, haloethoxy, halopropoxy, haloisopropoxy, halobutoxy, haloisobutoxy, sec-butoxy halide, tert-butoxy halide, etc.), C<com:Sub>3< / com:Sub>-C<com:Sub>6< / com:Sub> cycloalkyl (preferably cyclopropyl, cyclopentyl, cyclohexyl, etc.), C<com:Sub>3< / com:Sub>-C<com:Sub>6< / com:Sub> cycloalkyl halide (preferably halocyclopropyl, halocyclopentyl, halocyclohexyl, etc.), and 3- to 10- membered heterocyclyl (preferably tetrahydrofuranyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, etc.); substitution of aromatic rings can be monosubstitution (e.g., ortho-, meta- or para- substitution), disubstitution or trisubstitution, etc.

[0158] The term "heteroaryl" as used herein refers to an unsaturated ring group of 5 to 12 ring atoms with a fully conjugated <com:Math><semantics>π<annotation encoding="application / x-tex">\pi< / annotation>< / semantics>< / com:Math>-electron system, equivalent to the above "aryl" with one or more carbons replaced by a heteroatom such as oxygen, nitrogen, sulfur, etc. Heteroaromatic rings can be monocyclic or dicyclic, i.e., formed by fusion of two rings. A particular heterocyclic ring (heteroaryl) can be: pyridinyl, pyrimidinyl, pyrazinyl, isoxazolyl, isothiazolyl, pyrazolyl, thiazolyl, oxazolyl, imidazolyl, etc. Heteroaryl may be unsubstituted or substituted. A substituent of a heteroaryl is selected from halogen, cyano, nitro, amino, hydroxy, <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> alkyl, <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> hydroxyalkyl, <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> alkoxy, <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> alkyl halide, <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> hydroxyalkyl halide, C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkoxy halide, C<com:Sub>3< / com:Sub>-C<com:Sub>6< / com:Sub> cycloalkyl, C<com:Sub>3< / com:Sub>-C<com:Sub>6< / com:Sub> cycloalkyl halide and 3- to 10-membered heterocyclyl.

[0159] The term "alkyl" as used herein refers to a saturated straight monovalent hydrocarbon chain radical having from one to six carbon atoms, or a saturated branched monovalent hydrocarbon chain radical having from three to six carbon atoms, preferably being methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, iso-pentyl, neopentyl, etc. Alkyl may be unsubstituted or monosubstituted or multi-substituted, and the substituents may be the same or different in a case of multi-substitution; the substituent of alkyl is selected from halogen, nitro, hydroxy, C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkyl, C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkoxy, C<com:Sub>3< / com:Sub>-C<com:Sub>10< / com:Sub> cycloalkyl, alkoxycarbonyl, alkylthio, alkylsulfonyl, alkyamide, hydroxyalkylamide, sulfonamide, 3- to 10- membered heterocyclyl, or amino or mono- or multi-substituted amino, wherein substituents of amino may be the same or different, selected from H, C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkyl, C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> hydroxyalkyl, C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkoxy, C<com:Sub>3< / com:Sub>-C<com:Sub>10< / com:Sub> cycloalkyl, and 3- to 10-membered heterocyclyl.

[0160] The term "hydroxyalkyl" used herein refers to -alkyl-OH, wherein the alkyl is as above defined. Embodiments of "hydroxyalkyl" as used herein include, but are not limited to, hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxyisopropyl, etc. "Hydroxyalkyl" also includes substituted hydroxyalkyl, the substituent thereof may be halogen, amino, hydroxy, C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkyl, C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> hydroxyalkyl, C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkoxy or C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> cycloalkyl.

[0161] The term "alkoxy" used herein refers to -O-alkyl group, wherein the alkyl is as above defined. Embodiments of "alkoxy" as used herein include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy and tert-butoxy. "alkoxy" also includes substituted alkoxy, the substituent thereof may be halogen, amino, hydroxy, C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkyl, C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> hydroxyalkyl, C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkoxy or C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> cycloalkyl.

[0162] The term "cycloalkyl" used herein refers to a monocyclic or polycyclic (two monocyclic rings chemically bonded or bridged or spiro or fused), cyclically saturated, monovalent hydrocarbon group having 3 to 10 carbon atoms, preferably cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc., wherein one or two carbon atoms can be replaced by an oxo group. The cycloalkyl can be unsubstituted or substituted, the substituent thereof being selected from halogen, nitro, hydroxy, C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkyl, C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> hydroxyalkyl, C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkoxy, C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkyl halide, C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> hydroxyalkyl halide, C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkoxy halide, C<com:Sub>3< / com:Sub>-C<com:Sub>6< / com:Sub> cycloalkyl, C<com:Sub>3< / com:Sub>-C<com:Sub>6< / com:Sub> cycloalkyl halide, alkoxycarbonyl, alkylthio, alkylsulfonyl, alkyamide, hydroxyalkylamide, sulfonamide, 3- to 10- membered heterocyclyl, or amino or mono- or multi-substituted amino, wherein substituents of amino may be the same or different, selected from H, <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>C6<annotation encoding="application / x-tex">C_6< / annotation>< / semantics>< / com:Math> alkyl, <com:Math><semantics>C1<annotation encoding="application / x-tex">C_1< / annotation>< / semantics>< / com:Math>- C<com:Sub>6< / com:Sub> hydroxyalkyl, C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkoxy, C<com:Sub>3< / com:Sub>-C<com:Sub>10< / com:Sub> cycloalkyl, and 3- to 10-membered heterocyclyl.

[0163] The term "heterocyclyl" used herein refers to a monocyclic or polycyclic (two monocyclic rings chemically bonded or bridged or spiro or fused) cyclyl having 3 to 10 ring atoms, one or more heteroatoms being selected from N, O and S, including but not limited to [Image disponible dans le document PDF, Image available in the PDF document] cycloalkyl can be unsubstituted or substituted, the substituent thereof being selected from halogen, nitro, hydroxy, C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkyl, C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> hydroxyalkyl, C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkoxy, C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkyl halide, C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> hydroxyalkyl halide, C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkoxy halide, C<com:Sub>3< / com:Sub>-C<com:Sub>6< / com:Sub> cycloalkyl, C<com:Sub>3< / com:Sub>-C<com:Sub>6< / com:Sub> cycloalkyl halide, alkoxycarbonyl, alkylthio, alkylsulfonyl, alkyamide, hydroxyalkylamide, sulfonamide, 3- to 10- membered heterocyclyl, or amino, or mono- or multi-substituted amino, wherein substituents of amino may be the same or different, selected from II, C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkyl, C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> hydroxyalkyl, C<com:Sub>1< / com:Sub>-C<com:Sub>6< / com:Sub> alkoxy, C<com:Sub>3< / com:Sub>-C<com:Sub>10< / com:Sub> cycloalkyl and 3- to 10-membered heterocyclyl.

[0164] The term "alkenyl" used herein refers to branched or unbranched hydrocarbon chain of 2 to 10 carbon atoms having at least one double bond (i.e., C<com:Sub>2< / com:Sub>-C<com:Sub>10< / com:Sub> alkenyl), including but not limited to vinyl, allyl, but-1-enyl, pent-1-enyl, pent-1,4-di-alkenyl, etc. Alkenyl can be substituted by one or more substituents, the substituents are independently alkyl, alkyl halide, alkoxy, alkoxy halide, hydroxyalkyl, hydroxyalkyl halide, cycloalkyl, cycloalkyl halide, heterocycloalkyl, aryl, heteroaryl, hydroxy, halogen, cyano or nitro.

[0165] The term "alkinyl" used herein refers to branched or unbranched hydrocarbon chain composed of carbon and hydrogen atoms, containing at least one triple bond and 2 to 10 carbon atoms (i.e., C<com:Sub>2< / com:Sub>-C<com:Sub>10< / com:Sub> alkinyl), including but not limited to ethynyl, propargyl, butynyl, pentynyl and hexynyl, etc. Alkinyl can be substituted by one or more substituents, the substituents are independently alkyl, alkyl halide, alkoxy, alkoxy halide, hydroxyalkyl, hydroxyalkyl halide, cycloalkyl, cycloalkyl halide, heterocycloalkyl, aryl, heteroaryl, hydroxy, halogen, cyano or nitro.

[0166] The term "halogen" used herein refers to fluorine, chlorine, bromine or iodine, preferably fluorine, chlorine or bromine.

[0167] The term "halide" used herein refers to having a same atom or different atoms substituted by halogen, substitution being one time or a plurality of times, such as di- substitution or tri-substitution.

[0168] The term "alkyl halide" used herein refers to alkyl group substituted by halogen (preferably fluorine, chlorine, bromine or iodine), wherein the alkyl is as above defined. "Alkyl halide" may be substituted by halogen once or a plurality of times.

[0169] The term "hydroxyalkyl halide" used herein refers to hydroxyalkyl group substituted by halogen (preferably fluorine, chlorine, bromine or iodine), wherein the hydroxyalkyl is as above defined. "Hydroxyalkyl halide" may be substituted by halogen once or a plurality of times.

[0170] The term "haloalkoxy" used herein refers to alkoxy group substituted by halogen (preferably fluorine, chlorine, bromine or iodine), wherein the alkoxy is as above defined. "haloalkoxy" may be substituted by halogen once or a plurality of times.

[0171] The term "cycloalkyl halide" used herein refers to cycloalkyl group substituted by halogen (preferably fluorine, chlorine, bromine or iodine), wherein the cycloalkyl is as above defined. "Cycloalkyl halide" may be substituted by halogen once or a plurality of times.

[0172] The term "m" as used herein is preferably 1 or 2; such as it being two same <com:Math><semantics>R1<annotation encoding="application / x-tex">R_1< / annotation>< / semantics>< / com:Math> or two different R<com:Sub>1< / com:Sub>. Take R<com:Sub>1< / com:Sub> being F or Cl as an example, when m is 2, it may be Cl at two different substitution positions, or F at two different substitution positions, or a combination of Cl and F at two different substitution positions.

[0173] The term "n" as used herein is preferably absent or 1 or 2.

[0174] The term "o" as used herein is preferably 1, 2 or 3.

[0175] In the compounds of formula I, formulas II-A~II-C, formulas III-A~III-C, formulas IV-A~IV-C and formulas V-A~V-C, C denoted with * is chiral and the compound has an R- configuration or S-configuration, or a mixture of R- and S-configurations. Further preferably, the compound has an S-configuration.

[0176] The term "solvate" as used herein refers to a variable stoichiometry complex formed by a solute (e.g., a compound of general formulas I to V-A~V-C of the present invention) and a solvent. For the purposes of the present invention, the solvent must not interfere with the biological activity of the solute. Examples of suitable solvents include, but are not limited to, water, methanol, ethanol, and acetic acid. Preferably, the solvent used is a pharmaceutically acceptable solvent. The pharmaceutically acceptable solvent as a suitable solvent includes, but is not limited to, water, ethanol, and acetic acid. Further preferably, the solvent used is water.

[0177] In the present invention, a salt of the compound herein may be prepared by using methods known to those skilled in the art. The salt may be an organic acid salt, an inorganic acid salt, etc. The organic acid salt comprises fumarate, oxalate, malate, lactate, camphorsulfonate, p-toluenesulfonate, methanesulfonate, citrate, succinate, tartarate, maleate, benzoate, etc.; the inorganic acid salt comprises hydrohalate, sulfate, phosphate, nitrate, etc. For example, reaction with a lower alkyl sulfonic acid, such as methanesulfonic acid (methanesulphonic acid), trifluoromethanesulfonic acid, etc., may yield methanesulfonate (methanesulphonate), trifluoromethanesulfonate; reaction with aryl sulfonic acid, such as benzene sulfonic acid or p-toluenesulfonic acid, etc. may yield benzene sulfonate or p- toluenesulfonate; reaction with organic carboxylic acid, such as acctic acid, fumaric acid, tartaric acid, oxalic acid, maleic acid, malic acid, succinic acid or citric acid, etc. may yield a corresponding salt; reaction with amino acids, such as glutamic acid or aspartic acid may yield glutamate or aspartate. Reaction with an inorganic acid such as hydrohalic acid (e.g. hydrofluoric acid, hydrobromic acid, hydroiodic acid, hydrochloric acid), nitric acid, carbonic acid, sulfuric acid or phosphoric acid, may also yield a corresponding salt thereof.

[0178] The second purpose of the present invention is to provide a pharmaceutical composition comprising one or more of the compounds of any one above-mentioned technical scheme. The pharmaceutical composition herein may be composed of one or more of the compounds in any one of the above technical schemes with other compounds, or composed of one or more of the compounds in any one of the above technical schemes

[0179] In another aspect, provided in the present invention is an application of the compounds disclosed herein of general formulas I to general formulas V-A~V-C, and the stereoisomers thereof or stereoisomer mixtures thereof, or the pharmaceutically acceptable salts or solvates thereof in inhibiting the activity of the case in kinase <com:Math><semantics>1ϵ<annotation encoding="application / x-tex">1\epsilon< / annotation>< / semantics>< / com:Math> (Ck1<com:Math><semantics>ϵ<annotation encoding="application / x-tex">\epsilon< / annotation>< / semantics>< / com:Math>), or treating diseases, disorders or conditions that benefit from the inhibition of the activity of the casein kinase <com:Math><semantics>lϵ<annotation encoding="application / x-tex">l\epsilon< / annotation>< / semantics>< / com:Math> (CK<com:Math><semantics>lϵ<annotation encoding="application / x-tex">l\epsilon< / annotation>< / semantics>< / com:Math>).

[0180] In a further preferable scheme, provided by the present invention is a method for inhibition of the activity of the casein kinase <com:Math><semantics>1ϵ<annotation encoding="application / x-tex">1\epsilon< / annotation>< / semantics>< / com:Math> (CK1<com:Math><semantics>ϵ<annotation encoding="application / x-tex">\epsilon< / annotation>< / semantics>< / com:Math>) of a subject by administration of a composition comprising at lease on compound of a therapeutically effective amount in the subject in need, wherein the compound has a structural formula of compounds of general formulas I to V-A~V-C.

[0181] In certain embodiments, the subject in need has cancer, the cancer comprising hematologic neoplasms and solid tumors, such as B-cell lymphoma, multiple myeloma, leukemia, lung cancer, breast cancer, prostate cancer, bladder cancer, ovarian cancer, pancreatic cancer, sarcoma, colon cancer, kidney cancer, melanoma, and brain tumors.

[0182] In a further embodiment, the subject in need has B-cell lymphoma, such as diffuse large B-cell lymphoma, follicular lymphoma, chronic lymphocytic lymphoma, chronic lymphocytic leukemia, B-cell prolymphocytic leukemia, lymphoplasmacytic lymphoma / Waldenström macroglobulinemia, splenic marginal zone lymphoma, plasma cell myeloma, plasmacytoma, extranodal marginal zone B-cell lymphoma, nodal marginal zone B-cell lymphoma, mantle cell lymphoma, mediastinal (thymus) large B-cell lymphoma, intravascular large B-cell lymphoma, primary effusion lymphoma, Burkitt's lymphoma / leukemia or pulmonary lymphomatoid granulomatosis.

[0183] In a further embodiment, the subject in need has an autoimmune disease, such as rheumatoid arthritis, systemic lupus erythematosus, multiple sclerosis, inflammatory bowel disease, Crohn's disease, colitis, autoimmune hemolytic anemia, ankylosing spondylitis, pemphigus, urticaria, asthma, optic neuritis, psoriasis, chronic obstructive airway disease, dermatitis and baldness.

[0184] In a further embodiment, the subject in need has T-cell lymphoma, such as peripheral T-cell lymphoma, anaplastic large cell lymphoma, extranodal NK / T-cell lymphoma, cutaneous peripheral T-cell lymphoma, T-cell prolymphocytic leukemia, angioimmunoblastic T-cell lymphoma, adult T-cell leukemia / lymphoma, hepatosplenic T-cell lymphoma, intestinal T-cell lymphoma, breast implant-associated anaplastic large cell lymphoma, T-cell large granular lymphocytic leukemia.

[0185] Further provided in the present invention is an application of the compound or the pharmaceutically applicable salt thereof in preparation of a CK1\varepsilon inhibitor, particularly an application in preparation of drugs for treating proliferative diseases. The proliferative disease comprises cancer. In other words, further provided in the present invention is an application of the compounds of general formulas I to V-A~V-C, the stereoisomers or stereoisomer mixtures thereof, or the pharmaceutically acceptable salts or solvates thereof in treating proliferative diseases (such as cancers), used alone or in combination with other drugs. The antineoplastic drug which can be used in combination with the compound herein or the pharmaceutically applicable salt thereof includes, but is not limited to, at least one of the following types: mitotic inhibitors (e.g., vinblastine, vindisine and vinorelbine), tubulin proteolysis inhibitors (e.g., taxol), alkylation reagents (e.g., cisplatin, carboplatin, cyclophosphamide, chlorambucil and bendamustine), antimetabolites (e.g., 5-fluorouracil, tegafur, methotrexate, cytarabine and hydroxycarbamide), insertable antibiotics (e.g., adriamycin, mitomycin and bleomycin), enzymes (e.g., aspartate aminotransferase), topoisomerase inhibitors (e.g., etoposide and camptothecin), biological response modifiers (e.g., interferon), immunomodulators (e.g., lenalidomide), BTK inhibitors (e.g., ibrutinib, acalabrutinib), Bcl-2 inhibitors (e.g., Venetoclax), anti-CD20 monoclonal antibodies (e.g., rituximab, ofatumumab and obinutuzumab), mTOR inhibitors (e.g., rapamune and AZD8055), mTORC1 inhibitors (e.g., everolimus and temsirolimus), AKT inhibitors (e.g., MK-2206 and GSD690693), PI3K inhibitors (e.g., Idelalisib and Duvelisib), proteasome inhibitors (e.g., bortezomib, carfilzomib and ixazomib), EGFR inhibitors (e.g., erlotinib, gefitinib and osimertinib), VEGFR inhibitors (sorafenib, cabozantinib, ravatinib, levatinib, apatinib), CDK inhibitors (e.g., paboxinib, reboxinib) and PD-1 / PD-L1 inhibitors (pembrolizumab, nivolumab, toripalimab, sintilimab).

[0186] The present invention also relates to a method for treating proliferative diseases by using the compound or pharmaceutical composition in combination with radiation treatment Techniques for radiation treatment administration are known in the art and can be used in the combination therapy of the present application.

[0187] The compound of the present invention can also be used in combination with one or more steroidal anti-inflammatory drugs, non-steroidal anti-inflammatory drugs or immunoselective anti-inflammatory derivatives, and combinations thereof.

[0188] The applicant has reported in prior patent literature (Patent Application No. 201710998664.5) a BTK irreversible inhibitor and optical isomer thereof or pharmaceutically acceptable salt or solvate thereof, of which 17a is a representative compound. Through further studies of the compound of the present application, we have found that the compound herein showed excellent synergistic anti-tumor activities in combination with a BTK inhibitor.

[0189] As demonstrated by experiments, the compound of the present invention has a potent inhibitory activity on in-vitro CK1ε kinase and in-vitro anti-tumor cells, showed good anti- tumor activity in an OCI-LY10 subcutaneous xenogeneic model when used alone, and excellent synergistic anti-tumor activity in combination with a BTK inhibitor, and the compound of the present invention has good pharmacokinetics properties and may be used alone or in combination with other drugs to treat diseases, disorders or conditions that benefit from the inhibition of the activity of the casein kinase 1ε, the diseases, disorders or conditions comprising tumors, autoimmune diseases, etc. DESCRIPTION OF THE EMBODIMENTS

[0190] The implementability of the present invention is illustrated hereinafter by means of embodiments. It should be understood by those skilled in the art that modifications or replacements of corresponding technical features based on the prior art still fall within the scope of protection claimed by the present invention. Embodiment 1: Synthesis of Intermediate 4 [Image disponible dans le document PDF, Image available in the PDF document]

[0191] Step 1: Compound 1 (117mmol) and 40ml of DMF were placed in a 250ml reaction flask to dissolve with stirring at 0°C, then propionyl chloride (121mmol) was slowly dripped with a large amount of white solid precipitated after dripping. After 1h of the reaction at the maintained temperature, 200ml of water was added with stirring for 1h, then filtered and dried to obtain a solid product, Intermediate 2.

[0192] Step 2: Intermediate 2 (10mmol), H<com:Sub>2< / com:Sub>N-B (20mmol) and 20ml of methylbenzene were placed in a 100ml reaction flask. PCl3<com:Sub>3< / com:Sub> (10mmol) was dripped with stirring at room temperature, then the temperature was elevated to 110°C for reflux overnight. Once the reaction was completed as detected by the TLC test, ethyl acetate and water were used to extract. After drying and concentration, the crude product was purified by silica gel column chromatography to obtain Intermediate 3. [01<com:S>9< / com:S>3] Step 3: Intermediate 3 (8mmol), sodium acetate (11.2mmol) and 20ml of glacial acetic acid were placed in a 100ml reaction flask with stirring at room temperature, and then bromine (8mmol) was slowly dripped. The reaction continued after dripping for about 3h at room temperature till basically completed as detected by the TLC test; sodium thiosulfate aqueous solution was added with stirring for 15min, then ethyl acetate and water were used to extract. After drying and concentration, the crude product was purified by silica gel column chromatography to obtain Intermediate 4.

[0194] The following compounds were prepared using the synthetic route and the method for the above Intermediate 4:

[0195] For ease of description, Compound 1 is defined as: R<com:Sub>1< / com:Sub>-substituted 2-aminobenzoic acid; [Image disponible dans le document PDF, Image available in the PDF document]

[0196] Br 4a (R<com:Sub>1< / com:Sub> in Compound 1 is H, B in H<com:Sub>2< / com:Sub>N-B is phenyl)

[0197] 2-(1-bromoethyl)-3-phenylquinazolin-4(3H)-one; 3-step reaction with 55% overall yield; LC-MS (ESI-MS): <com:Math><semantics>329[M+H]+<annotation encoding="application / x-tex">329[M+H]^+< / annotation>< / semantics>< / com:Math>; <com:Sup>1< / com:Sup>H NMR (400 MHz, CDCl<com:Sub>3< / com:Sub>) <com:Math><semantics>δ<annotation encoding="application / x-tex">\delta< / annotation>< / semantics>< / com:Math> 8.34 (d, <com:Math><semantics>J=8.0<annotation encoding="application / x-tex">J=8.0< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>7.91−7.76<annotation encoding="application / x-tex">7.91-7.76< / annotation>< / semantics>< / com:Math> (m, 2H), <com:Math><semantics>7.71−7.49<annotation encoding="application / x-tex">7.71-7.49< / annotation>< / semantics>< / com:Math> (m, 5H), <com:Math><semantics>7.22<annotation encoding="application / x-tex">7.22< / annotation>< / semantics>< / com:Math> (dt, <com:Math><semantics>J=7.3<annotation encoding="application / x-tex">J = 7.3< / annotation>< / semantics>< / com:Math>, 2.9 Hz, 1H), <com:Math><semantics>4.62<annotation encoding="application / x-tex">4.62< / annotation>< / semantics>< / com:Math> (q, <com:Math><semantics>J=6.7<annotation encoding="application / x-tex">J = 6.7< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>2.10(d,J=6.7Hz,3H)<annotation encoding="application / x-tex">2.10 (d, J = 6.7 Hz, 3H)< / annotation>< / semantics>< / com:Math>. [Image disponible dans le document PDF, Image available in the PDF document]

[0198] 4b (R<com:Sub>1< / com:Sub> in Compound 1 is substituted Cl at 6-position; B in H<com:Sub>2< / com:Sub>N-B is phenyl)

[0199] 2-(1-bromoethyl)-5-chloro-3-phenylquinazolin-4(3H)-one, 3-step reaction with 49% overall yield; LC-MS (ESI-MS): <com:Math><semantics>363[M+H]+<annotation encoding="application / x-tex">363[M+H]^+< / annotation>< / semantics>< / com:Math>; <com:Sup>1< / com:Sup>H NMR (500 MHz, CDCl<com:Sub>3< / com:Sub>) <com:Math><semantics>δ<annotation encoding="application / x-tex">\delta< / annotation>< / semantics>< / com:Math> 7.71 (dd, J = 8.2, 1.2 Hz, 1H), 7.64 (dd, <com:Math><semantics>J−14.4<annotation encoding="application / x-tex">J - 14.4< / annotation>< / semantics>< / com:Math>, 6.3 Hz, 1H), 7.62 – 7.48 (m, 5H), 7.20 – 7.13 (m, 1H), <com:Math><semantics>4.52<annotation encoding="application / x-tex">4.52< / annotation>< / semantics>< / com:Math> (q, J – <com:Math><semantics>6.7<annotation encoding="application / x-tex">6.7< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>2.03<annotation encoding="application / x-tex">2.03< / annotation>< / semantics>< / com:Math> (t, J – <com:Math><semantics>7.2<annotation encoding="application / x-tex">7.2< / annotation>< / semantics>< / com:Math> Hz, 3H). [Image disponible dans le document PDF, Image available in the PDF document]

[0200] 4c (R<com:Sub>1< / com:Sub> in Compound 1 is F substituted at 6-position; B in H<com:Sub>2< / com:Sub>N-B is phenyl)

[0201] 2-(1-bromoethyl)-5-fluoro-3-phenylquinazolin-4(3H)-one; 3-step reaction with 28% overall yield; LC-MS (ESI-MS): <com:Math><semantics>347[M+H]+<annotation encoding="application / x-tex">347[M+H]^+< / annotation>< / semantics>< / com:Math>; <com:Sup>1< / com:Sup>H NMR (500 MHz, CDCl<com:Sub>3< / com:Sub>) <com:Math><semantics>δ<annotation encoding="application / x-tex">\delta< / annotation>< / semantics>< / com:Math> 7.73 (dd, J = 13.5, 8.1 Hz, 1H), 7.57 (dd, <com:Math><semantics>J=22.5<annotation encoding="application / x-tex">J = 22.5< / annotation>< / semantics>< / com:Math>, 6.9 Hz, 5H), 7.16 (t, <com:Math><semantics>J=9.0<annotation encoding="application / x-tex">J = 9.0< / annotation>< / semantics>< / com:Math> Hz, 2H), 4.52 (q, <com:Math><semantics>J=6.6<annotation encoding="application / x-tex">J = 6.6< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>2.02<annotation encoding="application / x-tex">2.02< / annotation>< / semantics>< / com:Math> (d, <com:Math><semantics>J=6.7<annotation encoding="application / x-tex">J = 6.7< / annotation>< / semantics>< / com:Math> Hz, 3H). [Image disponible dans le document PDF, Image available in the PDF document]

[0202] Вr 4d (R<com:Sub>1< / com:Sub> in Compound 1 is F substituted at 5-position; B in H<com:Sub>2< / com:Sub>N-B is phenyl)

[0203] 2-(1-bromoethyl)-6-fluoro-3-phenylquinazolin-4(3H)-one; 3-step reaction with 39% overall yield; LC-MS (ESI-MS): 347 [M+H]<com:Sup>+< / com:Sup>. [Image disponible dans le document PDF, Image available in the PDF document]

[0204] 4e (R<com:Sub>1< / com:Sub> in Compound 1 is F substituted at 3-position and Cl substituted at 6-position; B in H<com:Sub>2< / com:Sub>N-B is phenyl)

[0205] 2-(1-bromoethyl)-5-chloro-8-fluoro-3-phenylquinazolin-4(3H)-one, 3-step reaction with 26% overall yield; LC-MS (ESI-MS): 381 [M+H]<com:Sup>+< / com:Sup>. [Image disponible dans le document PDF, Image available in the PDF document]

[0206] f (R<com:Sub>1< / com:Sub> in Compound 1 is F substituted at 3-position and F substituted at 6-position; B in H<com:Sub>2< / com:Sub>N-B is phenyl)

[0207] 2-(1-bromoethyl)-5,8-difluoro-3-phenylquinazolin-4(3H)-one, 3-step reaction with 20% overall yield; LC-MS (ESI-MS): 365 [M+H]<com:Sup>+< / com:Sup>. [Image disponible dans le document PDF, Image available in the PDF document]

[0208] <com:Math><semantics>4g<annotation encoding="application / x-tex">4g< / annotation>< / semantics>< / com:Math> (R<com:Sub>1< / com:Sub> in Compound 1 is H; B in H<com:Sub>2< / com:Sub>N-B is pyridin-3-yl)

[0209] 2-(1-bromoethyl)-3-(pyridin-3-yl)quinazolin-4(3H)-one, 3-step reaction with 36% overall yield; LC-MS (ESI-MS): 330 [M+H]<com:Sup>+< / com:Sup>. [Image disponible dans le document PDF, Image available in the PDF document]

[0210] 4h (R<com:Sub>1< / com:Sub> in Compound 1 is Cl substituted at 6-position; B in H<com:Sub>2< / com:Sub>N- B is pyridin-3-yl)

[0211] 2-(1-bromoethyl)-5-chloro-3-(pyridin-3-yl)quinazolin-4(3H)-one, 3-step reaction with 33% overall yield; LC-MS (ESI-MS): 364 [M+H]<com:Sup>+< / com:Sup>. [Image disponible dans le document PDF, Image available in the PDF document]

[0212] Br 4i (R<com:Sub>1< / com:Sub> in Compound 1 is Cl substituted at 6-position; B in H<com:Sub>2< / com:Sub>N-B is 5-fluoropyridin-3-yl)

[0213] 2-(1-bromoethyl)-5-chloro-3-(5-fluoropyridin-3-yl)quinazoline-4(3H)-one, 3-step reaction with 35% overall yield; LC-MS (ESI-MS): 382 [M+H]<com:Sup>+< / com:Sup>. [Image disponible dans le document PDF, Image available in the PDF document]

[0214] 4j (R<com:Sub>1< / com:Sub> in Compound 1 is H; B in H<com:Sub>2< / com:Sub>N-B is 3-methoxypropyl)

[0215] 2-(1-bromoethyl)-3-(3-methoxypropyl)quinazolin-4(3H)-one, 3-step reaction with 45% overall yield; LC-MS (ESI-MS): 325 [M+H]<com:Sup>+< / com:Sup>. [Image disponible dans le document PDF, Image available in the PDF document]

[0216] 4k (R<com:Sub>1< / com:Sub> in Compound 1 is Cl substituted at 6-position; B in H<com:Sub>2< / com:Sub>N-B is 3-methoxypropyl)

[0217] 2-(1-bromoethyl)-5-chloro-3-(3-methoxypropyl)quinazolin-4(3H)-one, 3-step reaction with 46% overall yield; LC-MS (ESI-MS): 359 [M+II]<com:Sup>+< / com:Sup>. [Image disponible dans le document PDF, Image available in the PDF document]

[0218] 4l (R<com:Sub>1< / com:Sub> in Compound 1 is Cl substituted at 6-position; B in H<com:Sub>2< / com:Sub>N- B is 2-(tetrahydrofuran-2-yl)ethyl)

[0219] 2-(1-bromoethyl)-5-chloro-3-(2-(tetrahydrofuran-2-yl)ethyl)quinazolin-4(3H)-one, 3-step reaction with 30% overall yield; LC-MS (ESI-MS): 385 [M+H]<com:Sup>+< / com:Sup>. [Image disponible dans le document PDF, Image available in the PDF document]

[0220] 4m (R<com:Sub>1< / com:Sub> in Compound 1 is Cl substituted at 6-position; B in H<com:Sub>2< / com:Sub>N-B is cyclohexyl)

[0221] 2-(1-bromoethyl)-5-chloro-3-cyclohexylquinazolin-4(3H)-one, 3-step reaction with 18% overall yield; LC-MS (ESI-MS): 369 [M+H]<com:Sup>+< / com:Sup>. [Image disponible dans le document PDF, Image available in the PDF document]

[0222] 4n (R<com:Sub>1< / com:Sub> in Compound 1 is Cl substituted at 6-position; B in H<com:Sub>2< / com:Sub>N-B is pyridin-1-yl)

[0223] 2-(1-bromoethyl)-5-chloro-3-(pyridin-1-yl)quinazolin-4(3H)-one, 3-step reaction with 36% overall yield; LC-MS (ESI-MS): 370 [M+H]<com:Sup>+< / com:Sup>. [Image disponible dans le document PDF, Image available in the PDF document]

[0224] Вr 4o (R<com:Sub>1< / com:Sub> in Compound 1 is Cl substituted at 6-position; B in H<com:Sub>2< / com:Sub>N-B is morpholin-4-yl)

[0225] 2-(1-bromoethyl)-5-chloro-3-morpholinquinazolin-4(3H)-one, 3-step reaction with 40% overall yield; LC-MS (ESI-MS): 372 [M+H]<com:Sup>+< / com:Sup>. [Image disponible dans le document PDF, Image available in the PDF document]

[0226] 4p (<com:Math><semantics>R1<annotation encoding="application / x-tex">R_1< / annotation>< / semantics>< / com:Math> in Compound 1 is Cl substituted at 6-position; B in <com:Math><semantics>H2N1<annotation encoding="application / x-tex">H_2N_1< / annotation>< / semantics>< / com:Math> B is 4-boc-piperazinyl)

[0227] 4-(2-(1-bromoethyl)-5-chloro-4-oxoquinazolin-3(4H)-yl)piperazine-1-carboxylic acid tert-butyl ester, 3-step reaction with 28% overall yield; LC-MS (ESI-MS): 471 [M+H]<com:Sup>+< / com:Sup>. [Image disponible dans le document PDF, Image available in the PDF document]

[0228] 4q (R<com:Sub>1< / com:Sub> in Compound 1 is Cl substituted at 6-position; B in H<com:Sub>2< / com:Sub>N-B is 3-trifluoromethyl-4-boc-piperazinyl)

[0229] 4-(2-(1-bromoethyl)-5-chloro-4-oxoquinazolin-3(4H)-yl)-2- (trifluoromethyl)piperazine-1-carboxylic acid tert-butyl ester, 3-step reaction with 36% overall yield; LC-MS (ESI-MS): 539 [M+H]<com:Sup>+< / com:Sup>. [Image disponible dans le document PDF, Image available in the PDF document]

[0230] 4r (R<com:Sub>1< / com:Sub> in Compound 1 is Cl substituted at 6-position; B in H<com:Sub>2< / com:Sub>N-B is 3,3-dimethyl-4-boc-piperazinyl)

[0231] 4-(2-(1-bromoethyl)-5-chloro-4-oxoquinazolin-3(4H)-yl)-2,2-dimethylpiperazine- 1-carboxylic acid tert-butyl ester, 3-step reaction with 31% overall yield; LC-MS (ESI-MS): 499 [M+H]<com:Sup>+< / com:Sup>. [Image disponible dans le document PDF, Image available in the PDF document]

[0232] 4s (R<com:Sub>1< / com:Sub> in Compound 1 is Cl substituted at 6-position; B in H<com:Sub>2< / com:Sub>N-B is N-Boc-pyridin-4-yl)

[0233] 4-(2-(1-bromoethyl)-5-chloro-4-oxoquinazolin-3(4H)-yl)pyridin-1-carboxylic acid tert-butyl ester, 3-step reaction with 15% overall yield; LC-MS (ESI-MS): 470 [M+H]<com:Sup>+< / com:Sup>. Embodiment 2: Synthesis of Intermediate 9 [Image disponible dans le document PDF, Image available in the PDF document]

[0234] Step 1: Compound 5 (5g, 19mmol), triphenylmethyl chloride (10.7g, 38mmol), potassium carbonate (10.6g, 76mmol) and 50ml of DMF were placed separately in a 100ml reaction flask and reacted for about 24h at 80°C. Once the reaction was completed as detected by the TLC test, the temperature was lowered to room temperature, and ethyl acetate and water were added. Insoluble substances were removed by filtration, and the ethyl acetate layer was separated. The crude product concentrated under reduced pressure was purified by silical gel column chromatography to obtain 6.75g of the product, Intermediate 6, with a yield of 73%.

[0235] Step 2: Intermediate 6 (18mmol), compound 7, potassium carbonate (54mmol), tetrakis(triphenylphosphine)palladium(0) (0.9mmol), 135ml of dioxane and 45ml of water were placed separately in a 500ml reaction flask. The temperature was elevated to 100°C with stirring to react for 14h and brought to room temperature when the reaction was completed as detected by the TLC test. The reaction liquid was filtered with diatomite, and the filter cake was rinsed with ethyl acetate. The recovered filtrate was extracted with 100ml of water and 300ml of ethyl acetate. The ethyl acetate layer was separated, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to obtain the product, Intermediate 8.

[0236] Step 3: Intermediate 8 (3.85mmol) and 30ml of dichloromethane were placed separately in a 100ml reaction flask, then mixed with 1.15ml of TFA (15.4mmol) and 3.1ml of triisopropylsilane (15.4mmol) to react for 18h at room temperature. When the reaction was completed as detected by the TLC test, the solvent was directly evaporated with a rotary evaporator under reduced pressure till almost driness, then triturated with 20ml of MTBE (methyl tert-butyl ether) for 2h at room temperature and filtered to obtain the target product, Intermediate 9.

[0237] The following compounds were prepared using the synthetic route and the method of Intermediate 9:

[0238] The positions of carbon atoms on the conjugate ring in Compound 7 are defined as follows. [Image disponible dans le document PDF, Image available in the PDF document]

[0239] [Image disponible dans le document PDF, Image available in the PDF document]

[0240] 9a (in Compound 7: R<com:Sub>2< / com:Sub> is methyl, X is CF; and R<com:Sub>3< / com:Sub> is H)

[0241] 3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine, 3-step reaction with 55% overall yield; LC-MS (ESI-MS): 278 [M+H]<com:Sup>+< / com:Sup>. [Image disponible dans le document PDF, Image available in the PDF document]

[0242] 9b (in Compound 7: R<com:Sub>2< / com:Sub> is isopropyl, X is CF, and R<com:Sub>3< / com:Sub> is H)

[0243] 3-(2,3-difluoro-4-isopropoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine, 3-step reaction with 42% overall yield; LC-MS (ESI-MS): 306 [M+H]<com:Sup>+< / com:Sup>. [Image disponible dans le document PDF, Image available in the PDF document]

[0244] 9c (Compound 7 was replaced with 3-fluoro-4- methoxyphenylboronic acid)

[0245] 3-(3-fluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine, 3-step reaction with 58% overall yield; LC-MS (ESI-MS): 260 [M+H]<com:Sup>+< / com:Sup>. [Image disponible dans le document PDF, Image available in the PDF document]

[0246] 9d (Compound 7 was replaced with 3-fluoro-4- isopropoxyphenylboronic acid)

[0247] 3-(3-fluoro-4-isopropoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine, 3-step reaction with 55% overall yield; LC-MS (ESI-MS): 288 [M+H]<com:Sup>+< / com:Sup>. Embodiment 4: Synthesis of Intermediate 10 [Image disponible dans le document PDF, Image available in the PDF document]

[0248] 3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (5.2g, 20mmol), potassium carbonate (4.2g, 30mmol) and 100ml of DMF were placed separately in a 500ml reaction flask, and then mixed with Compound 4 (8.6g, 26mmol) with stirring at room temperature to react for about 12h at room temperate. When the reaction was completed as detected by the TLC test, ethyl acetate and water were added to extract and then water was used to wash for 3 times. The ethyl acetate layer was separated, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was then purified by silica gel column chromatography to obtain the white solid product, Intermediate 10. [Image disponible dans le document PDF, Image available in the PDF document]

[0249] 10a (Compound 4 was 2-(1-bromoethyl)-3-phenylquinazolin-4(3H)- one, i.e., <com:Math><semantics>R1=H<annotation encoding="application / x-tex">R_1=H< / annotation>< / semantics>< / com:Math>)

[0250] 2-(1-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-3- phenylquinazolin-4(3H)-one; 83% yield. LC-MS (ESI-MS): 510 [M+H]<com:Sup>+< / com:Sup>. Embodiment 5: Synthesis of Intermediates (R)-12a~(R)-12d [Image disponible dans le document PDF, Image available in the PDF document]

[0251] Step 1: Methyl (R)-(+)-lactate (20g, 192mmol) and 400ml of methyl tert-butyl ether (MTBE) were placed in a 1000ml reaction flask, and then mixed with benzyl bromide (36g, 211mmol) and silver oxide (44g, 192mmol) with stirring at room temperature to react for about 20h at room temperature till the reaction was completed as detected by the TLC test. The reaction liquid was filtered with diatomite and the filter cake with rinsed with MTBE. The filtrate was then concentrated under reduced pressure and purified by silica gel column chromatography to obtain 29.8g of a colorless oily product methyl (R) 2- (benzyloxy)propanoate with a yield of 80%. Chiral assay ee%: 99.0%, chiral-HPLC being performed using a normal-phase CHIRALCEL<com:Sup>TM< / com:Sup> OD-H column with a retention time of 10.12min (retention time for the enantiomer was 8.07min). <com:Sup>1< / com:Sup>H NMR (500 MHz, CDCl<com:Sub>3< / com:Sub>) δ <com:Math><semantics>7.44−7.27<annotation encoding="application / x-tex">7.44-7.27< / annotation>< / semantics>< / com:Math> (m, 5H), <com:Math><semantics>4.69<annotation encoding="application / x-tex">4.69< / annotation>< / semantics>< / com:Math> (d, <com:Math><semantics>J=11.7<annotation encoding="application / x-tex">J = 11.7< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>4.46<annotation encoding="application / x-tex">4.46< / annotation>< / semantics>< / com:Math> (d, <com:Math><semantics>J=11.7<annotation encoding="application / x-tex">J = 11.7< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>4.07<annotation encoding="application / x-tex">4.07< / annotation>< / semantics>< / com:Math> (q, <com:Math><semantics>J=6.8<annotation encoding="application / x-tex">J = 6.8< / annotation>< / semantics>< / com:Math> Hz, 1H), 3.76 (s, 3H), 1.44 (d, <com:Math><semantics>J=6.9<annotation encoding="application / x-tex">J = 6.9< / annotation>< / semantics>< / com:Math> Hz, 3H).

[0252] Step 2: Methyl (R)-2-(benzyloxy)propanoate (11g, 57mmol) and 100ml of methanol were placed in a reaction flask, and then mixed with lithium hydroxide (2.1g, 85mmol) with stirring to react for about 5h at room temperature till the reaction was completed as detected by the TLC test. Then 0.1N aqueous hydrochloric acid was added to neutralized to neutrality. Most of the methanol was removed by concentration under reduced pressure, then ethyl acetate and water were added to extract. The ethyl acetate layer was separated, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 8.0g of a colorless oily product (R)- 2-(benzyloxy)propionic acid with a yield of 78%. <com:Sup>1< / com:Sup>H NMR (500 MHz, CDCl<com:Sub>3< / com:Sub>) δ 7.45–7.28 (m, 5H), <com:Math><semantics>4.72<annotation encoding="application / x-tex">4.72< / annotation>< / semantics>< / com:Math> (d, <com:Math><semantics>J=11.6<annotation encoding="application / x-tex">J = 11.6< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>4.53<annotation encoding="application / x-tex">4.53< / annotation>< / semantics>< / com:Math> (d, <com:Math><semantics>J=11.6<annotation encoding="application / x-tex">J = 11.6< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>4.12<annotation encoding="application / x-tex">4.12< / annotation>< / semantics>< / com:Math> (q, <com:Math><semantics>J=6.9<annotation encoding="application / x-tex">J = 6.9< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>1.50<annotation encoding="application / x-tex">1.50< / annotation>< / semantics>< / com:Math> <com:Math><semantics>(d,J=6.9 Hz,3H).<annotation encoding="application / x-tex">(d, J = 6.9 \text{ Hz}, 3\text{H}).< / annotation>< / semantics>< / com:Math>

[0253] Step 3: Substituted 2-amino-benzoic acid (19mmol), (R)-2-(benzyloxy)propionic acid (4.0g, 22mmol), triphenyl phosphite (23g, 76mmol) and 50ml of pyridine were placed in a reaction flask, then the reaction mixture was heated to 60°C and reacted for 2h with stirring. The B-NH2 (38mmol) was added to the reaction system, which was then heated to 70°C and reacted for about 12h with stirring till the reaction was completed as detected by the TLC test, and then brought to room temperature and concentrated under reduced pressure to remove most of the pyridine. Ethyl acetate was added, and washed with 2M aqueous potassium bisulfate for 3 times. The ethyl acetate layer was separated, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain a white solid product, Intermediate (R)-11.

[0254] Step 4: (R)-11 (5.4 mmol) and 50ml of dichloromethane were added in a reaction flask, cooled to -10°C, and 28ml of 2M boron tribromide in dichloromethane solution was dripped. The mixture was then heated to room temperature after dripping to react for about 2h till the reaction was completed as detected by the TLC test. The reaction flask was placed in a ice bath and water was added to quench the reaction, and then dichloromethane was added to extract. The dichloromethane layer was separated, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain a white solid product, (R)-12.

[0255] The following compounds were prepared by using the synthetic route and the method of Intermediate (R)-12: [Image disponible dans le document PDF, Image available in the PDF document]

[0256] ŌН (R)-12a (R<com:Sub>1< / com:Sub> in substituted 2-amino-benzoic acid is H; B in B-NH<com:Sub>2< / com:Sub> is phenyl)

[0257] (R)-2-(1-hydroxyethyl)-3-phenylquinazolin-4(3H)-one, 1.4g with 97% yield in the last step. LC-MS (ESI-MS): 267 [M+H]<com:Sup>+< / com:Sup>. [Image disponible dans le document PDF, Image available in the PDF document]

[0258] (R)-12b (R<com:Sub>1< / com:Sub> in substituted 2-amino-benzoic acid is Cl substituted at 6-position; B in B-NH<com:Sub>2< / com:Sub> is phenyl)

[0259] (R)-5-chloro-2-(1-hydroxyethyl)-3-phenylquinazolin-4(3H)-one, 1.6g with 98% yield in the last step. Chiral assay ee%: 98.5%, chiral-HPLC being performed using a reversed- phase CHIRALPAK<com:Sup>TM< / com:Sup> IG-3 column with a retention time of 52.7min. <com:Sup>1< / com:Sup>H NMR (500 MHz, CDCl<com:Sub>3< / com:Sub>) <com:Math><semantics>δ<annotation encoding="application / x-tex">\delta< / annotation>< / semantics>< / com:Math> 7.70–7.62 (m, 2H), 7.60–7.52 (m, 3H), 7.52–7.49 (m, 1H), 7.32 (dd, J = 7.5, <com:Math><semantics>2.2 Hz<annotation encoding="application / x-tex">2.2 \text{ Hz}< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>1H<annotation encoding="application / x-tex">1\text{H}< / annotation>< / semantics>< / com:Math>), <com:Math><semantics>7.27−7.23 (m, 1H)<annotation encoding="application / x-tex">7.27-7.23 \text{ (m, 1H)}< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>4.45 (q, J=6.4 Hz<annotation encoding="application / x-tex">4.45 \text{ (q, J} = 6.4 \text{ Hz}< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>1H<annotation encoding="application / x-tex">1\text{H}< / annotation>< / semantics>< / com:Math>), <com:Math><semantics>1.24 (d, J=6.4 Hz<annotation encoding="application / x-tex">1.24 \text{ (d, J} = 6.4 \text{ Hz}< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>3H<annotation encoding="application / x-tex">3\text{H}< / annotation>< / semantics>< / com:Math>). LC-MS <com:Math><semantics>(ESI−MS):301[M+H]+.<annotation encoding="application / x-tex">(ESI-MS): 301 [M+H]^+.< / annotation>< / semantics>< / com:Math> [Image disponible dans le document PDF, Image available in the PDF document]

[0260] (R)-12c (R<com:Sub>1< / com:Sub> in substituted 2-amino-benzoic acid is F substituted at 6- position; B in B-NII<com:Sub>2< / com:Sub> is phenyl)

[0261] (R)-5-fluoro-2-(1-hydroxyethyl)-3-phenylquinazolin-4(3II)-one, 1.5g with 98% yield in the last step. LC-MS (ESI-MS): 285 [M+H]<com:Sup>+< / com:Sup>. [Image disponible dans le document PDF, Image available in the PDF document]

[0262] (R)-12d (R<com:Sub>1< / com:Sub> in substituted 2-amino-benzoic acid is Cl substituted at 6-position; B in B-NH<com:Sub>2< / com:Sub> is pyridin-3-yl)

[0263] (R)-5-chloro-2-(1-hydroxyethyl)-3-(pyridin-3-yl)quinazolin-4(3H)-one, 1.5g with 92% yield in the last step. LC-MS (ESI-MS): 302 [M+H]<com:Sup>+< / com:Sup>. Embodiment 6: Synthesis of Intermediates (S)-12a~(S)-12d [Image disponible dans le document PDF, Image available in the PDF document]

[0264] By referring to the preparation method of Intermediate (R)-12 of Embodiment 5, the following compounds were prepared using methyl (S)-(-)-lactate as the starting material: [Image disponible dans le document PDF, Image available in the PDF document]

[0265] ŌΗ (S)-12a (R<com:Sub>1< / com:Sub> in substituted 2-amino-benzoic acid is H; B in B-NH<com:Sub>2< / com:Sub> is phenyl)

[0266] (S)-2-(1-hydroxyethyl)-3-phenylquinazolin-4(3H)-one, 1.4g with 97% yield in the last step. LC-MS (ESI-MS): 267 [M+H]<com:Sup>+< / com:Sup>. [Image disponible dans le document PDF, Image available in the PDF document]

[0267] ŌΗ (S)-12b (R<com:Sub>1< / com:Sub> in substituted 2-amino-benzoic acid is Cl substituted at 6-position; B in B-NH<com:Sub>2< / com:Sub> is phenyl)

[0268] (S)-5-chloro-2-(1-hydroxyethyl)-3-phenylquinazolin-4(3H)-one, 1.5g with 93% yield in the last step. Chiral assay ee%: 98.5%, chiral-HPLC being performed using a reversed-phase CIIIRALPAK IG-3 column with a retention time of 43.7min. LC-MS (ESI- MS): <com:Math><semantics>301[M+II]+<annotation encoding="application / x-tex">301 [M+II]^+< / annotation>< / semantics>< / com:Math>. [Image disponible dans le document PDF, Image available in the PDF document]

[0269] ŌΗ (S)-12c (R<com:Sub>1< / com:Sub> in substituted 2-amino-benzoic acid is F substituted at 6- position; B in B-NH<com:Sub>2< / com:Sub> is phenyl)

[0270] (S)-5-fluoro-2-(1-hydroxyethyl)-3-phenylquinazolin-4(3H)-one, 1.4g with 91% yield in the last step. LC-MS (ESI-MS): 285 [M+H]<com:Sup>+< / com:Sup>. [Image disponible dans le document PDF, Image available in the PDF document]

[0271] ŌН (S)-12d (R<com:Sub>1< / com:Sub> in substituted 2-amino-benzoic acid is Cl substituted at 6-position; B in B-NH<com:Sub>2< / com:Sub> is pyridin-3-yl)

[0272] (S)-5-chloro-2-(1-hydroxyethyl)-3-(pyridin-3-yl)quinazolin-4(3H)-one, 1.5g with 92% yield in the last step. LC-MS (ESI-MS): 302 [M+H]<com:Sup>+< / com:Sup>. Embodiment 7: Synthesis of Target Compounds II-A-1~II-A-6 and II-A-12~II-A- 15 [Image disponible dans le document PDF, Image available in the PDF document]

[0273] Intermediate 10 (2.0mmol), intermediate 7 (3.0mmol), potassium carbonate (4.0mmol), tetrakis(triphenylphosphine)palladium(0) (0.02mmol) were mixed with the solvent, 9ml of dioxane and 3ml of water in a 50ml reaction flask, heated to 100°C with stirring and reacted for about 14h till the reaction was completed as detected by the TLC test, then brought to room temperature. The reaction liquid was filtered with diatomite and rinsed with ethyl acetate. The filtrate was extracted with 20ml of water and 50ml of ethyl acetate, then the ethyl acetate layer was separated, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to obtain a white solid product.

[0274] Target compound II-A-1 (R<com:Sub>2< / com:Sub>=CH<com:Sub>3< / com:Sub>, R<com:Sub>3< / com:Sub>=H, X=CF): white solid product, 0.76g, 72% yield. <com:Sup>1< / com:Sup>H NMR (500 MHz, CDCl<com:Sub>3< / com:Sub>) <com:Math><semantics>δ<annotation encoding="application / x-tex">\delta< / annotation>< / semantics>< / com:Math> 8.27 (d, <com:Math><semantics>J=7.9<annotation encoding="application / x-tex">J = 7.9< / annotation>< / semantics>< / com:Math> Hz, 1H), 8.04 (s, 1H), 7.89–7.72 (m, 2H), <com:Math><semantics>7.61−7.40<annotation encoding="application / x-tex">7.61-7.40< / annotation>< / semantics>< / com:Math> (m, 2H), <com:Math><semantics>7.31<annotation encoding="application / x-tex">7.31< / annotation>< / semantics>< / com:Math> (d, <com:Math><semantics>J=7.7<annotation encoding="application / x-tex">J=7.7< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>7.21<annotation encoding="application / x-tex">7.21< / annotation>< / semantics>< / com:Math> (t, <com:Math><semantics>J=7.5<annotation encoding="application / x-tex">J=7.5< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>7.16<annotation encoding="application / x-tex">7.16< / annotation>< / semantics>< / com:Math> (t, <com:Math><semantics>J=7.4<annotation encoding="application / x-tex">J=7.4< / annotation>< / semantics>< / com:Math> Hz, 1H), 6.90 (t, <com:Math><semantics>J=7.7<annotation encoding="application / x-tex">J = 7.7< / annotation>< / semantics>< / com:Math> Hz, 1H), 6.85 (t, <com:Math><semantics>J=7.9<annotation encoding="application / x-tex">J = 7.9< / annotation>< / semantics>< / com:Math> Hz, 1H), 6.54 (d, <com:Math><semantics>J=7.7<annotation encoding="application / x-tex">J = 7.7< / annotation>< / semantics>< / com:Math> Hz, 1H), 6.03 (q, <com:Math><semantics>J=<annotation encoding="application / x-tex">J =< / annotation>< / semantics>< / com:Math> 6.5 Hz, 1H), 5.45 (brs, 2H), 3.94 (s, 3H), 1.93 (d, <com:Math><semantics>J=6.7<annotation encoding="application / x-tex">J = 6.7< / annotation>< / semantics>< / com:Math> Hz, 3H). LC-MS (ESI-MS): 526 <com:Math><semantics>[M+H]+<annotation encoding="application / x-tex">[M+H]^{+}< / annotation>< / semantics>< / com:Math>.

[0275] Target compound II-A-2 (R<com:Sub>2< / com:Sub>=CH(CH<com:Sub>3< / com:Sub>)<com:Sub>2< / com:Sub>, R<com:Sub>3< / com:Sub>=H, X=CF): white solid product, 0.78g, 70% yield. LC-MS (ESI-MS): 554 [M+H]<com:Sup>+< / com:Sup>.

[0276] Target compound II-A-3 (R<com:Sub>2< / com:Sub>=CH<com:Sub>2< / com:Sub>CH<com:Sub>3< / com:Sub>, R<com:Sub>3< / com:Sub>=H, X=CF): white solid product, 0.68g, 63% yield. <com:Sup>1< / com:Sup>H NMR (500 MHz, CDCl<com:Sub>3< / com:Sub>) <com:Math><semantics>δ<annotation encoding="application / x-tex">\delta< / annotation>< / semantics>< / com:Math> 8.27 (d, <com:Math><semantics>J=7.8<annotation encoding="application / x-tex">J = 7.8< / annotation>< / semantics>< / com:Math> Hz, 1H), 8.04 (s, 1H), 7.92 – 7.73 <com:Math><semantics>(m,2H),7.49(dt,J=18.2,7.7Hz,2H),7.31(d,J=7.8Hz,1H),7.17(dt,J=14.7,7.3Hz,1.3Hz,1.4Hz,1.4Hz,1.4Hz,1.4Hz,1.4Hz,1.4Hz,1.4Hz,1.4Hz,1.4Hz,1.4Hz,1.4Hz,1.4Hz,1.4Hz,1.4Hz,1.4<annotation encoding="application / x-tex">(m, 2H), 7.49 (dt, J = 18.2, 7.7 Hz, 2H), 7.31 (d, J = 7.8 Hz, 1H), 7.17 (dt, J = 14.7, 7.3 Hz, 1.3 Hz, 1.4 Hz, 1.4 Hz, 1.4 Hz, 1.4 Hz, 1.4 Hz, 1.4 Hz, 1.4 Hz, 1.4 Hz, 1.4 Hz, 1.4 Hz, 1.4 Hz, 1.4 Hz, 1.4 Hz, 1.4 Hz, 1.4< / annotation>< / semantics>< / com:Math> 2H), 6.89 (t, <com:Math><semantics>J=7.6<annotation encoding="application / x-tex">J = 7.6< / annotation>< / semantics>< / com:Math> Hz, 1H), 6.83 (t, <com:Math><semantics>J=7.6<annotation encoding="application / x-tex">J = 7.6< / annotation>< / semantics>< / com:Math> Hz, 1H), 6.53 (d, <com:Math><semantics>J=7.9<annotation encoding="application / x-tex">J = 7.9< / annotation>< / semantics>< / com:Math> Hz, 1H), 6.03 (q, <com:Math><semantics>J=7.6<annotation encoding="application / x-tex">J = 7.6< / annotation>< / semantics>< / com:Math> Hz, 1H), 6.03 (q, <com:Math><semantics>J=7.6<annotation encoding="application / x-tex">J = 7.6< / annotation>< / semantics>< / com:Math> Hz, 1H), 6.03 (q, <com:Math><semantics>J=7.6<annotation encoding="application / x-tex">J = 7.6< / annotation>< / semantics>< / com:Math> Hz, 1H), 6.03 (q, <com:Math><semantics>J=7.6<annotation encoding="application / x-tex">J = 7.6< / annotation>< / semantics>< / com:Math> Hz, 1H), 6.03 (q, <com:Math><semantics>J=7.6<annotation encoding="application / x-tex">J = 7.6< / annotation>< / semantics>< / com:Math> H 6.7 Hz, 1H), 5.38 (brs, 2H), 4.16 (q, <com:Math><semantics>J=6.9<annotation encoding="application / x-tex">J = 6.9< / annotation>< / semantics>< / com:Math> Hz, 2H), 1.92 (d, <com:Math><semantics>J=6.7<annotation encoding="application / x-tex">J = 6.7< / annotation>< / semantics>< / com:Math> Hz, 3H), 1.48 (t, <com:Math><semantics>J=7.0<annotation encoding="application / x-tex">J = 7.0< / annotation>< / semantics>< / com:Math> Hz, 3H). LC-MS (ESI-MS): 540 [M+H]<com:Sup>+< / com:Sup>.

[0277] Target compound II-A-4 (R<com:Sub>2< / com:Sub>=cyclopropyl, R<com:Sub>3< / com:Sub>=H, X=CF): white solid product, 0.41g, 37% yield. <com:Sup>1< / com:Sup>H NMR (500 MHz, CDCl3) <com:Math><semantics>δ<annotation encoding="application / x-tex">\delta< / annotation>< / semantics>< / com:Math> 8.27 (d, <com:Math><semantics>J=7.9<annotation encoding="application / x-tex">J = 7.9< / annotation>< / semantics>< / com:Math> Hz, 1H), 8.03 (s, 1H), 7.80 <com:Math><semantics>(ddd,J=11.6,9.6,4.3 Hz,2H),7.55−7.45 (m, 2H),7.31 (d, J=7.3 Hz,1H),7.24−7.13 (m, 2H)<annotation encoding="application / x-tex">(ddd, J = 11.6, 9.6, 4.3 \text{ Hz}, 2\text{H}), 7.55 - 7.45 \text{ (m, 2H)}, 7.31 \text{ (d, } J = 7.3 \text{ Hz}, 1\text{H)}, 7.24 - 7.13 \text{ (m, 2H)}< / annotation>< / semantics>< / com:Math> 2H), 6.89 (t, <com:Math><semantics>J=7.7<annotation encoding="application / x-tex">J = 7.7< / annotation>< / semantics>< / com:Math> Hz, 1H), 6.53 (d, <com:Math><semantics>J=7.9<annotation encoding="application / x-tex">J = 7.9< / annotation>< / semantics>< / com:Math> Hz, 1H), 6.03 (q, <com:Math><semantics>J=6.7<annotation encoding="application / x-tex">J = 6.7< / annotation>< / semantics>< / com:Math> Hz, 1H), 5.49 (brs, 2H), 3.87 (ddd, <com:Math><semantics>J=9.0<annotation encoding="application / x-tex">J = 9.0< / annotation>< / semantics>< / com:Math>, 5.9, 3.1 Hz, 1H), 1.93 (t, <com:Math><semantics>J=6.0<annotation encoding="application / x-tex">J = 6.0< / annotation>< / semantics>< / com:Math> Hz, 3H), 0.89 – 0.82 (m, 4H). LC- MS (ESI-MS): 552 [M+H]<com:Sup>+< / com:Sup>.

[0278] Target compound II-A-5 (R<com:Sub>2< / com:Sub>=CH<com:Sub>3< / com:Sub>, R<com:Sub>3< / com:Sub>=H, X=CCl): white solid product, 0.44g, 41% yield. LC-MS (ESI-MS): 542 [M+H]<com:Sup>+< / com:Sup>.

[0279] Target compound II-A-6 (R<com:Sub>2< / com:Sub>=CH<com:Sub>3< / com:Sub>, R<com:Sub>3< / com:Sub>=H, X=CCN): white solid product, 0.42g, 39% yield. LC-MS (ESI-MS): 533 [M+H]<com:Sup>+< / com:Sup>.

[0280] Target compound II-A-12 (R<com:Sub>2< / com:Sub>=CH<com:Sub>3< / com:Sub>, R<com:Sub>3< / com:Sub>=6-F, X=CH): white solid product, 0.36g, 34% yield. <com:Sup>1< / com:Sup>H NMR (500 MHz, CDCl<com:Sub>3< / com:Sub>) <com:Math><semantics>δ<annotation encoding="application / x-tex">\delta< / annotation>< / semantics>< / com:Math> 8.27 (d, <com:Math><semantics>J=8.0<annotation encoding="application / x-tex">J = 8.0< / annotation>< / semantics>< / com:Math> Hz, 1H), 8.03 (s, 1H), 7.91 – 7.71 (m, 2H), <com:Math><semantics>7.63−7.41<annotation encoding="application / x-tex">7.63-7.41< / annotation>< / semantics>< / com:Math> (m, 2H), <com:Math><semantics>7.31<annotation encoding="application / x-tex">7.31< / annotation>< / semantics>< / com:Math> (d, <com:Math><semantics>J=7.6<annotation encoding="application / x-tex">J = 7.6< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>7.24−7.18<annotation encoding="application / x-tex">7.24-7.18< / annotation>< / semantics>< / com:Math> (m, 1H), <com:Math><semantics>7.15<annotation encoding="application / x-tex">7.15< / annotation>< / semantics>< / com:Math> (dd, <com:Math><semantics>J=14.5<annotation encoding="application / x-tex">J = 14.5< / annotation>< / semantics>< / com:Math>, 7.0 Hz, 1H), 6.89 (dd, <com:Math><semantics>J=11.1<annotation encoding="application / x-tex">J = 11.1< / annotation>< / semantics>< / com:Math>, 4.4 Hz, 1H), 6.85 (t, <com:Math><semantics>J=7.4<annotation encoding="application / x-tex">J = 7.4< / annotation>< / semantics>< / com:Math> Hz, 1H), 6.54 (d, <com:Math><semantics>J=8.1<annotation encoding="application / x-tex">J = 8.1< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>6.03<annotation encoding="application / x-tex">6.03< / annotation>< / semantics>< / com:Math> (q, <com:Math><semantics>J=6.7<annotation encoding="application / x-tex">J = 6.7< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>5.49<annotation encoding="application / x-tex">5.49< / annotation>< / semantics>< / com:Math> (brs, 2H), <com:Math><semantics>3.94<annotation encoding="application / x-tex">3.94< / annotation>< / semantics>< / com:Math> (s, 3H), <com:Math><semantics>1.92<annotation encoding="application / x-tex">1.92< / annotation>< / semantics>< / com:Math> (d, <com:Math><semantics>J=6.7<annotation encoding="application / x-tex">J = 6.7< / annotation>< / semantics>< / com:Math> Hz, 3H). LC-MS (ESI- MS): 526 [M+H]<com:Sup>+< / com:Sup>.

[0281] Target compound II-A-13 (<com:Math><semantics>R2<annotation encoding="application / x-tex">R_2< / annotation>< / semantics>< / com:Math>=CH<com:Sub>3< / com:Sub>, <com:Math><semantics>R3<annotation encoding="application / x-tex">R_3< / annotation>< / semantics>< / com:Math>=H, X=N): white solid product, 0.75g, 74% yield. LC-MS (ESI-MS): 509 [M+H]<com:Sup>+< / com:Sup>.

[0282] Target compound II-A-14 (R<com:Sub>2< / com:Sub>=CH(CH<com:Sub>3< / com:Sub>)<com:Sub>2< / com:Sub>, R<com:Sub>3< / com:Sub>=H, X=N): white solid product, <com:Math><semantics>0.72g,67%<annotation encoding="application / x-tex">0.72g, 67\%< / annotation>< / semantics>< / com:Math> yield. <com:Sup>1< / com:Sup>H NMR (500 MHz, CDCl<com:Sub>3< / com:Sub>) <com:Math><semantics>δ<annotation encoding="application / x-tex">\delta< / annotation>< / semantics>< / com:Math> 8.27 (dd, <com:Math><semantics>J=7.9,0.9<annotation encoding="application / x-tex">J = 7.9, 0.9< / annotation>< / semantics>< / com:Math> Hz, 1H), 8.03 (s, 1H), <com:Math><semantics>7.89−7.75<annotation encoding="application / x-tex">7.89 - 7.75< / annotation>< / semantics>< / com:Math> (m, 2H), <com:Math><semantics>7.54−7.49<annotation encoding="application / x-tex">7.54 - 7.49< / annotation>< / semantics>< / com:Math> (m, 1H), <com:Math><semantics>7.47<annotation encoding="application / x-tex">7.47< / annotation>< / semantics>< / com:Math> (td, <com:Math><semantics>J=7.7<annotation encoding="application / x-tex">J = 7.7< / annotation>< / semantics>< / com:Math>, 1.2 Hz, 1H), <com:Math><semantics>7.31<annotation encoding="application / x-tex">7.31< / annotation>< / semantics>< / com:Math> (d, <com:Math><semantics>J=7.9<annotation encoding="application / x-tex">J = 7.9< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>7.16<annotation encoding="application / x-tex">7.16< / annotation>< / semantics>< / com:Math> (t, <com:Math><semantics>J=7.5<annotation encoding="application / x-tex">J = 7.5< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>6.96−6.87<annotation encoding="application / x-tex">6.96 - 6.87< / annotation>< / semantics>< / com:Math> (m, 1H), <com:Math><semantics>6.67<annotation encoding="application / x-tex">6.67< / annotation>< / semantics>< / com:Math> (d, <com:Math><semantics>J=8.2<annotation encoding="application / x-tex">J = 8.2< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>6.51<annotation encoding="application / x-tex">6.51< / annotation>< / semantics>< / com:Math> (d, <com:Math><semantics>J=8.0<annotation encoding="application / x-tex">J = 8.0< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>6.02<annotation encoding="application / x-tex">6.02< / annotation>< / semantics>< / com:Math> (q, <com:Math><semantics>J=6.7<annotation encoding="application / x-tex">J = 6.7< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>5.60<annotation encoding="application / x-tex">5.60< / annotation>< / semantics>< / com:Math> (brs, 2H), <com:Math><semantics>5.33−5.22<annotation encoding="application / x-tex">5.33 - 5.22< / annotation>< / semantics>< / com:Math> (m, 1H), <com:Math><semantics>4.06<annotation encoding="application / x-tex">4.06< / annotation>< / semantics>< / com:Math> (t, <com:Math><semantics>J=6.7<annotation encoding="application / x-tex">J = 6.7< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>1.92<annotation encoding="application / x-tex">1.92< / annotation>< / semantics>< / com:Math> <com:Math><semantics>(d,J=6.8 Hz,3H),1.36(dd,J=6.2,3.0 Hz,6H).LC−MS(ESI−MS):537[M+H]+.<annotation encoding="application / x-tex">(d, J = 6.8 \text{ Hz}, 3H), 1.36 (dd, J = 6.2, 3.0 \text{ Hz}, 6H). LC-MS (ESI-MS): 537 [M+H]^+.< / annotation>< / semantics>< / com:Math>

[0283] Target compound II-A-15 (R<com:Sub>2< / com:Sub>=Ph, R<com:Sub>3< / com:Sub>=H, X=N): white solid product, 0.82g, 72% yield. <com:Sup>1< / com:Sup>H NMR (500 MHz, CDCl<com:Sub>3< / com:Sub>) <com:Math><semantics>δ<annotation encoding="application / x-tex">\delta< / annotation>< / semantics>< / com:Math> 8.27 (d, <com:Math><semantics>J=7.7<annotation encoding="application / x-tex">J = 7.7< / annotation>< / semantics>< / com:Math> Hz, 1H), 8.04 (s, 1H), 7.96 (t, <com:Math><semantics>J=8.8<annotation encoding="application / x-tex">J = 8.8< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.81 (dt, <com:Math><semantics>J=15.0<annotation encoding="application / x-tex">J = 15.0< / annotation>< / semantics>< / com:Math>, 7.4 Hz, 2H), 7.51 (t, <com:Math><semantics>J=6.9<annotation encoding="application / x-tex">J = 6.9< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.47 (t, <com:Math><semantics>J=7.9<annotation encoding="application / x-tex">J = 7.9< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.43 <com:Math><semantics>(t,J=7.9 Hz,2H),7.36−7.23 (m, 2H),7.16 (d, J=7.6 Hz,3H),6.90 (t, J=7.7 Hz,1H),<annotation encoding="application / x-tex">(t, J = 7.9 \text{ Hz}, 2H), 7.36 - 7.23 \text{ (m, 2H)}, 7.16 \text{ (d, } J = 7.6 \text{ Hz}, 3H), 6.90 \text{ (t, } J = 7.7 \text{ Hz}, 1H),< / annotation>< / semantics>< / com:Math> <com:Math><semantics>6.86<annotation encoding="application / x-tex">6.86< / annotation>< / semantics>< / com:Math> (d, <com:Math><semantics>J=8.1<annotation encoding="application / x-tex">J = 8.1< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>6.50<annotation encoding="application / x-tex">6.50< / annotation>< / semantics>< / com:Math> (d, <com:Math><semantics>J=7.8<annotation encoding="application / x-tex">J = 7.8< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>6.03<annotation encoding="application / x-tex">6.03< / annotation>< / semantics>< / com:Math> (q, <com:Math><semantics>J=6.6<annotation encoding="application / x-tex">J = 6.6< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>5.46<annotation encoding="application / x-tex">5.46< / annotation>< / semantics>< / com:Math> (brs, 2H), 1.92 (d, <com:Math><semantics>J=6.7<annotation encoding="application / x-tex">J = 6.7< / annotation>< / semantics>< / com:Math> Hz, 3H). LC-MS (ESI-MS): 571 [M+H]<com:Sup>+< / com:Sup>. Embodiment 8: Synthesis of Target Compounds II-A-7~II-A-11 [Image disponible dans le document PDF, Image available in the PDF document]

[0284] 3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (1.0mmol) and 5ml of DMF were placed separately in a 20ml reaction flask, and stirred to dissolve at room temperature, then mixed in turn with Intermediate 4 (1.1mmol) and potassium carbonate (2.0mmol), and heated to 40°C to react for about 15h. When the reaction was completed as detected by the TLC test, ethyl acetate was added to extract, and then washed with water. The ethyl acetate layer was separated, dried over anhydrous sodium sulphate and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain the target product.

[0285] Target compound II-A-7 (R<com:Sub>1< / com:Sub>=5-chloro): white solid product, 0.48g, 85% yield. <com:Sup>1< / com:Sup>H NMR (500 MHz, CDCl<com:Sub>3< / com:Sub>) <com:Math><semantics>δ<annotation encoding="application / x-tex">\delta< / annotation>< / semantics>< / com:Math> 8.05 (s, 1H), 7.73 (d, <com:Math><semantics>J=8.1<annotation encoding="application / x-tex">J = 8.1< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.64 (t, <com:Math><semantics>J=8.0<annotation encoding="application / x-tex">J = 8.0< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.51 (d, <com:Math><semantics>J=7.8<annotation encoding="application / x-tex">J = 7.8< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.47 (t, <com:Math><semantics>J=7.6<annotation encoding="application / x-tex">J = 7.6< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.29 (d, <com:Math><semantics>J=7.9<annotation encoding="application / x-tex">J = 7.9< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.22 (dd, <com:Math><semantics>J=11.8<annotation encoding="application / x-tex">J = 11.8< / annotation>< / semantics>< / com:Math>, 4.4 Hz, 1H), 7.16 (t, <com:Math><semantics>J=7.4<annotation encoding="application / x-tex">J = 7.4< / annotation>< / semantics>< / com:Math> Hz, 1H), 6.93 – 6.84 (m, 2H), 6.53 (d, <com:Math><semantics>J=8.0<annotation encoding="application / x-tex">J = 8.0< / annotation>< / semantics>< / com:Math> Hz, 1H), 6.00 (q, <com:Math><semantics>J=8.0<annotation encoding="application / x-tex">J = 8.0< / annotation>< / semantics>< / com:Math> Hz, 1H), 6.00 (q, <com:Math><semantics>J=8.0<annotation encoding="application / x-tex">J = 8.0< / annotation>< / semantics>< / com:Math> Hz, 1H), 6.00 (q, <com:Math><semantics>J=8.0<annotation encoding="application / x-tex">J = 8.0< / annotation>< / semantics>< / com:Math> Hz, 1H), 6.00 (q, <com:Math><semantics>J=8.0<annotation encoding="application / x-tex">J = 8.0< / annotation>< / semantics>< / com:Math> Hz, 1H), 6.00 (q, <com:Math><semantics>J=8.0<annotation encoding="application / x-tex">J = 8.0< / annotation>< / semantics>< / com:Math> <com:Math><semantics>=6.7 Hz,1H,5.33 (brs, 2H),3.96 (s, 3H),1.90 (d, J=6.7 Hz, 3H).LC−MS (ESI-MS):560 (s. 3H)<annotation encoding="application / x-tex">= 6.7 \text{ Hz}, 1\text{H}, 5.33 \text{ (brs, 2H)}, 3.96 \text{ (s, 3H)}, 1.90 \text{ (d, } J = 6.7 \text{ Hz, 3H)}. LC-MS \text{ (ESI-MS)}: 560 \text{ (s. 3H)}< / annotation>< / semantics>< / com:Math> <com:Math><semantics>[M+H]+<annotation encoding="application / x-tex">[M+H]^{+}< / annotation>< / semantics>< / com:Math>.

[0286] Target compound Π-A-8 (R<com:Sub>1< / com:Sub>=5-fluoro): white solid product, 0.36g, 66% yield. <com:Sup>1< / com:Sup>H NMR (500 MHz, CDCl<com:Sub>3< / com:Sub>) <com:Math><semantics>δ<annotation encoding="application / x-tex">\delta< / annotation>< / semantics>< / com:Math> 8.04 (s, 1H), 7.71 (dd, <com:Math><semantics>J=13.5<annotation encoding="application / x-tex">J = 13.5< / annotation>< / semantics>< / com:Math>, 8.1 Hz, 1H), 7.61 (d, <com:Math><semantics>J=8.2<annotation encoding="application / x-tex">J = 8.2< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.47 (t, <com:Math><semantics>J=7.6<annotation encoding="application / x-tex">J = 7.6< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.29 (d, <com:Math><semantics>J=8.1<annotation encoding="application / x-tex">J = 8.1< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.21 (t, <com:Math><semantics>J=7.9<annotation encoding="application / x-tex">J = 7.9< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.15 (t, <com:Math><semantics>J=7.6<annotation encoding="application / x-tex">J = 7.6< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.15 (t, <com:Math><semantics>J=7.6<annotation encoding="application / x-tex">J = 7.6< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.15 (t, <com:Math><semantics>J=7.6<annotation encoding="application / x-tex">J = 7.6< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.15 (t, <com:Math><semantics>J=7.6<annotation encoding="application / x-tex">J = 7.6< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.15 (t, <com:Math><semantics>J=7.6<annotation encoding="application / x-tex">J = 7.6< / annotation>< / semantics>< / com:Math> H 9.3 Hz, 2H), 6.87 (dd, <com:Math><semantics>J=17.1<annotation encoding="application / x-tex">J = 17.1< / annotation>< / semantics>< / com:Math>, 8.6 Hz, 2H), 6.52 (d, <com:Math><semantics>J=8.0<annotation encoding="application / x-tex">J = 8.0< / annotation>< / semantics>< / com:Math> Hz, 1H), 6.00 (q, <com:Math><semantics>J=6.6<annotation encoding="application / x-tex">J = 6.6< / annotation>< / semantics>< / com:Math> Hz, 1H), 5.31 (brs, 2H), 3.95 (s, 3H), 1.90 (d, <com:Math><semantics>J=6.7<annotation encoding="application / x-tex">J = 6.7< / annotation>< / semantics>< / com:Math> Hz, 3H). LC-MS (ESI-MS): 544 [M+H]<com:Sup>+< / com:Sup>.

[0287] Target compound II-A-9 (R<com:Sub>1< / com:Sub>=6-fluoro): white solid product, <com:Math><semantics>0.38g<annotation encoding="application / x-tex">0.38g< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>70%<annotation encoding="application / x-tex">70\%< / annotation>< / semantics>< / com:Math> yield. LC-MS (ESI-MS): 544 [M+H]<com:Sup>+< / com:Sup>.

[0288] Target compound II-A-10 (R<com:Sub>1< / com:Sub>=5-chloro-8-fluoro): white solid product, 0.34g, 58% yield. LC-MS (ESI-MS): 578 [M+H]<com:Sup>+< / com:Sup>.

[0289] Target compound II-A-11 (R<com:Sub>1< / com:Sub>=5,8-difluoro): white solid product, 0.37g, 65% yield. LC-MS (ESI-MS): 562 [M+H]<com:Sup>+< / com:Sup>. Embodiment 9: Synthesis of Target Compound II-A-16 [Image disponible dans le document PDF, Image available in the PDF document]

[0290] 3-(2,3-difluoro-4-isopropoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (9b, 1.0mmol) and 5ml of DMF were placed separately in a 20ml reaction flask and stirred to dissolve at room temperature, then mixed in turn with Intermediate 4c (1.1mmol) and potassium carbonate (2.0mmol), and heated to 40°C to react for about 15h. When the reaction was completed as detected by the TLC test, ethyl acetate was added to extract and washed with water. The ethyl acetate layer was separated, dried over anhydrous sodium sulphate and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 0.36g of the target product II-A-16 with a yield of 63%. <com:Sup>1< / com:Sup>H NMR (500 MHz, DMSO) δ 7.87 (brs, 2H), 7.60 (brs, 2H), 7.47–7.28 (m, 2H), 7.24–7.02 (m, 3H), 6.79 (brs, 1H), 6.31 (brs, 1H), 5.90 (brs, 1H), 4.71 (brs, 1H), 1.71 (s, 3H), 1.31 (s, 6H). LC- MS (ESI-MS): 572 [M+H]<com:Sup>+< / com:Sup>. Example 10: Synthesis of Target Compounds II-B-1~II-B-3 [Image disponible dans le document PDF, Image available in the PDF document]

[0291] 3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (9a, 1.0mmol) was placed in a 20ml reaction flask with 5ml of DMF, and stirred to dissolve at room temperature, then mixed in turn with Intermediate 4 (1.1mmol) and potassium carbonate (2.0mmol), and heated to 40°C to react for about 15h. When the reaction was completed as detected by the TLC test, ethyl acetate was added to extracted, and washed with water. The ethyl acetate layer was separated, dried over anhydrous sodium sulphate and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain the target product.

[0292] Target product II-B-1 (R<com:Sub>1< / com:Sub>=H, R<com:Sub>5< / com:Sub>=H): white solid product, 0.37g, 71% yield. LC- MS (ESI-MS): 527 [M+H]<com:Sup>+< / com:Sup>.

[0293] Target product II-B-2 (R<com:Sub>1< / com:Sub>=5-chloro, R<com:Sub>5< / com:Sub>=II): white solid product, 0.36g, 65% yield. <com:Sup>1< / com:Sup>H NMR (500 MHz, CDCl<com:Sub>3< / com:Sub>) <com:Math><semantics>δ<annotation encoding="application / x-tex">\delta< / annotation>< / semantics>< / com:Math> 8.58 (d, <com:Math><semantics>J=2.3<annotation encoding="application / x-tex">J = 2.3< / annotation>< / semantics>< / com:Math> Hz, 0.5H), 8.39 (d, <com:Math><semantics>J=3.6<annotation encoding="application / x-tex">J = 3.6< / annotation>< / semantics>< / com:Math> Hz, 0.5H), 8.30 (d, <com:Math><semantics>J=4.3 Hz,0.5H,8.08 (d, J=11.6 Hz,1H),7.81−7.72 (m, 1.5H),7.67 (td, J=8.1,3.9 Hz,<annotation encoding="application / x-tex">J = 4.3 \text{ Hz}, 0.5\text{H}, 8.08 \text{ (d, } J = 11.6 \text{ Hz}, 1\text{H}), 7.81 - 7.72 \text{ (m, 1.5H)}, 7.67 \text{ (td, } J = 8.1, 3.9 \text{ Hz},< / annotation>< / semantics>< / com:Math> 1.5H), <com:Math><semantics>7.58−7.50<annotation encoding="application / x-tex">7.58 - 7.50< / annotation>< / semantics>< / com:Math> (m, 1H), <com:Math><semantics>7.38<annotation encoding="application / x-tex">7.38< / annotation>< / semantics>< / com:Math> (dd, <com:Math><semantics>J=8.0<annotation encoding="application / x-tex">J = 8.0< / annotation>< / semantics>< / com:Math>, 4.9 Hz, 0.5H), <com:Math><semantics>7.29−7.23<annotation encoding="application / x-tex">7.29 - 7.23< / annotation>< / semantics>< / com:Math> (m, 0.5H), <com:Math><semantics>7.20<annotation encoding="application / x-tex">7.20< / annotation>< / semantics>< / com:Math> (t, <com:Math><semantics>J=8.0<annotation encoding="application / x-tex">J = 8.0< / annotation>< / semantics>< / com:Math>) <com:Math><semantics>=7.3 Hz,0.5H,6.92−6.79 (m, 2H),5.99 (q, J=6.7 Hz,0.5H),5.90 (q, J=6.8 Hz,0.5H),<annotation encoding="application / x-tex">= 7.3 \text{ Hz}, 0.5 \text{H}, 6.92 - 6.79 \text{ (m, 2H)}, 5.99 \text{ (q, } J = 6.7 \text{ Hz}, 0.5 \text{H)}, 5.90 \text{ (q, } J = 6.8 \text{ Hz}, 0.5 \text{H)},< / annotation>< / semantics>< / com:Math> 5.39 (brs, 2H), 3.95 (d, <com:Math><semantics>J=2.0<annotation encoding="application / x-tex">J = 2.0< / annotation>< / semantics>< / com:Math> Hz, 3H), 1.91 (t, <com:Math><semantics>J=6.1<annotation encoding="application / x-tex">J = 6.1< / annotation>< / semantics>< / com:Math> Hz, 3H). LC-MS (ESI-MS): 561 <com:Math><semantics>[M+H]+<annotation encoding="application / x-tex">[M+H]^{+}< / annotation>< / semantics>< / com:Math>.

[0294] Target product II-B-3 (R<com:Sub>1< / com:Sub>=5-chloro, R<com:Sub>5< / com:Sub>=5-fluoro): white solid product, 0.35g, 60% yield. <com:Sup>1< / com:Sup>H NMR (500 MHz, CDCl<com:Sub>3< / com:Sub>) <com:Math><semantics>δ<annotation encoding="application / x-tex">\delta< / annotation>< / semantics>< / com:Math> 8.43 (s, 0.6H), 8.26 (d, <com:Math><semantics>J=2.6<annotation encoding="application / x-tex">J = 2.6< / annotation>< / semantics>< / com:Math> Hz, 0.6H), 8.16 (d, <com:Math><semantics>J=2.6<annotation encoding="application / x-tex">J = 2.6< / annotation>< / semantics>< / com:Math> Hz, 0.6H), 8.16 (d, <com:Math><semantics>J=2.6<annotation encoding="application / x-tex">J = 2.6< / annotation>< / semantics>< / com:Math> Hz, 0.6H), 8.16 (d, <com:Math><semantics>J=2.6<annotation encoding="application / x-tex">J = 2.6< / annotation>< / semantics>< / com:Math> Hz, 0.6H), 8.16 (d, <com:Math><semantics>J=2.6<annotation encoding="application / x-tex">J = 2.6< / annotation>< / semantics>< / com:Math> Hz, 0.6H), 8.16 (d, <com:Math><semantics>J=2.<annotation encoding="application / x-tex">J = 2.< / annotation>< / semantics>< / com:Math> <com:Math><semantics>2.5 Hz,0.6H,8.12 (s, 1H),7.77 (d, J=8.1 Hz, 1H),7.69 (td, J=8.0,1.3 Hz, 1H),7.63−7.52<annotation encoding="application / x-tex">2.5 \text{ Hz}, 0.6 \text{H}, 8.12 \text{ (s, 1H)}, 7.77 \text{ (d, } J = 8.1 \text{ Hz, 1H)}, 7.69 \text{ (td, } J = 8.0, 1.3 \text{ Hz, 1H)}, 7.63-7.52< / annotation>< / semantics>< / com:Math> <com:Math><semantics>(m,1.4H),7.46−7.41(m,0.4H),7.25−7.18(m,1H),6.88(dd,<math>J=14.2,7.0Hz,1H),6.69−1.00Hz<annotation encoding="application / x-tex">(m, 1.4H), 7.46-7.41 (m, 0.4H), 7.25-7.18 (m, 1H), 6.88 (dd, J = 14.2, 7.0 Hz, 1H), 6.69-1.00 Hz< / annotation>< / semantics>< / com:Math> 6.57 (m, 0.7H), 5.97 (dq, <com:Math><semantics>J=24.5<annotation encoding="application / x-tex">J = 24.5< / annotation>< / semantics>< / com:Math>, 6.7 Hz, 1.3H), 5.33 (d, <com:Math><semantics>J=24.5<annotation encoding="application / x-tex">J = 24.5< / annotation>< / semantics>< / com:Math> Hz, 2H), 3.96 (d, <com:Math><semantics>J=3.3<annotation encoding="application / x-tex">J = 3.3< / annotation>< / semantics>< / com:Math> Hz, 3H), 2.11–1.76 (m, 3H). LC-MS (ESI-MS): 579 [M+H]<com:Sup>+< / com:Sup>. Embodiment 11: Synthesis of Target Compounds II-C-1, II-C-5 and II-C-6 [Image disponible dans le document PDF, Image available in the PDF document]

[0295] 3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (1.0mmol) was placed in a 20ml reaction flask with 5ml of DMF, and stirred to dissolve at room temperature, then mixed in turn with Intermediate 4 (1.1mmol) and potassium carbonate (2.0mmol), and heated to 40°C to react for about 15h. When the reaction was completed as detected by the TLC test, ethyl acetate was added to extract, and washed with water. The ethyl acetate layer was separated, dried with anhydrous sodium sulphate and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain the target product.

[0296] Target product II-C-1 (Y=CH, Z=CH<com:Sub>2< / com:Sub>): white solid product, 0.41g, 72% yield. <com:Sup>1< / com:Sup>H NMR (500 MHz, CDCl<com:Sub>3< / com:Sub>) <com:Math><semantics>δ<annotation encoding="application / x-tex">\delta< / annotation>< / semantics>< / com:Math> 8.46 (s, 1H), 7.65 (d, <com:Math><semantics>J=8.0<annotation encoding="application / x-tex">J = 8.0< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.55 (t, <com:Math><semantics>J=8.0<annotation encoding="application / x-tex">J = 8.0< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.45 (d, <com:Math><semantics>J=7.7<annotation encoding="application / x-tex">J = 7.7< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.18 (t, <com:Math><semantics>J=7.4<annotation encoding="application / x-tex">J = 7.4< / annotation>< / semantics>< / com:Math> Hz, 1H), 6.84 (t, <com:Math><semantics>J=7.9<annotation encoding="application / x-tex">J = 7.9< / annotation>< / semantics>< / com:Math> Hz, 1H), 6.36 (q, <com:Math><semantics>J=6.5<annotation encoding="application / x-tex">J = 6.5< / annotation>< / semantics>< / com:Math> Hz, 1H), 5.66 (brs, 2H), <com:Math><semantics>4.00−3.86<annotation encoding="application / x-tex">4.00-3.86< / annotation>< / semantics>< / com:Math> (m, 1H), <com:Math><semantics>3.93<annotation encoding="application / x-tex">3.93< / annotation>< / semantics>< / com:Math> (s, 3H), <com:Math><semantics>2.73<annotation encoding="application / x-tex">2.73< / annotation>< / semantics>< / com:Math> (qd, <com:Math><semantics>J=12.5<annotation encoding="application / x-tex">J=12.5< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>3.4<annotation encoding="application / x-tex">3.4< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>2.47<annotation encoding="application / x-tex">2.47< / annotation>< / semantics>< / com:Math> (qd, <com:Math><semantics>J=12.7,3.4 Hz,1H,1.99 (d, J=6.5 Hz,3H),1.86−1.77 (m, 1H),1.66 (d, J=12.0 Hz,1H),<annotation encoding="application / x-tex">J = 12.7, 3.4 \text{ Hz}, 1\text{H}, 1.99 \text{ (d, } J = 6.5 \text{ Hz}, 3\text{H}), 1.86 - 1.77 \text{ (m, } 1\text{H}), 1.66 \text{ (d, } J = 12.0 \text{ Hz}, 1\text{H}),< / annotation>< / semantics>< / com:Math> 1.44 (d, <com:Math><semantics>J=12.8<annotation encoding="application / x-tex">J = 12.8< / annotation>< / semantics>< / com:Math> Hz, 1H), 1.37 (d, <com:Math><semantics>J=13.0<annotation encoding="application / x-tex">J = 13.0< / annotation>< / semantics>< / com:Math> Hz, 1H), 1.29 (dd, <com:Math><semantics>J=16.7<annotation encoding="application / x-tex">J = 16.7< / annotation>< / semantics>< / com:Math>, 6.7 Hz, 1H), 1.16– 1.05 (m, 1H), 0.43 (d, <com:Math><semantics>J=11.6<annotation encoding="application / x-tex">J = 11.6< / annotation>< / semantics>< / com:Math> Hz, 1H), 0.11 (dd, <com:Math><semantics>J=26.0<annotation encoding="application / x-tex">J = 26.0< / annotation>< / semantics>< / com:Math>, 13.0 Hz, 1H). LC-MS (ESI-MS): 566 [M+H]<com:Sup>+< / com:Sup>.

[0297] Target product II-C-5 (Y=N, Z=CH<com:Sub>2< / com:Sub>): white solid product, 0.40g, 71% yield. <com:Sup>1< / com:Sup>H NMR (500 MHz, CDCl<com:Sub>3< / com:Sub>) <com:Math><semantics>δ<annotation encoding="application / x-tex">\delta< / annotation>< / semantics>< / com:Math> 8.43 (s, 1H), 7.59 (dt, <com:Math><semantics>J=6.7,3.3<annotation encoding="application / x-tex">J = 6.7, 3.3< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.54 (t, <com:Math><semantics>J=7.9<annotation encoding="application / x-tex">J = 7.9< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.43 (dd, <com:Math><semantics>J=7.7<annotation encoding="application / x-tex">J = 7.7< / annotation>< / semantics>< / com:Math>, 1.1 Hz, 1H), 7.25–7.17 (m, 1H), 6.84 (dd, <com:Math><semantics>J=11.8<annotation encoding="application / x-tex">J = 11.8< / annotation>< / semantics>< / com:Math>, 4.3 Hz, 1H), 6.53 <com:Math><semantics>(q,J=6.9 Hz,1H),5.54 (brs, 2H),4.02−3.90 (m, 1H),3.94 (s, 3H),3.88−3.72 (m, 1H),<annotation encoding="application / x-tex">(q, J = 6.9 \text{ Hz}, 1\text{H}), 5.54 \text{ (brs, 2H)}, 4.02 - 3.90 \text{ (m, 1H)}, 3.94 \text{ (s, 3H)}, 3.88 - 3.72 \text{ (m, 1H)},< / annotation>< / semantics>< / com:Math> 2.98 (d, <com:Math><semantics>J=10.3<annotation encoding="application / x-tex">J = 10.3< / annotation>< / semantics>< / com:Math> Hz, 1H), 2.26 (d, <com:Math><semantics>J=9.9<annotation encoding="application / x-tex">J = 9.9< / annotation>< / semantics>< / com:Math> Hz, 1H), 1.95 (d, <com:Math><semantics>J=6.9<annotation encoding="application / x-tex">J = 6.9< / annotation>< / semantics>< / com:Math> Hz, 3H), 1.78–1.68 (m, 3H), <com:Math><semantics>1.58−1.45<annotation encoding="application / x-tex">1.58 - 1.45< / annotation>< / semantics>< / com:Math> (m, 1H), <com:Math><semantics>1.29<annotation encoding="application / x-tex">1.29< / annotation>< / semantics>< / com:Math> (d, <com:Math><semantics>J=14.7<annotation encoding="application / x-tex">J = 14.7< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>1.22−1.06<annotation encoding="application / x-tex">1.22 - 1.06< / annotation>< / semantics>< / com:Math> (m, 1H), <com:Math><semantics>0.45<annotation encoding="application / x-tex">0.45< / annotation>< / semantics>< / com:Math> (dtd, <com:Math><semantics>J=12.9<annotation encoding="application / x-tex">J = 12.9< / annotation>< / semantics>< / com:Math>, 9.0, 4.0 Hz, 1H). LC-MS (ESI-MS): 567 [M+H]<com:Sup>+< / com:Sup>.

[0298] Target product II-C-6 (Y=N, Z=O): white solid product, 0.31g, 55% yield. <com:Sup>1< / com:Sup>H NMR (500 MHz, CDCl<com:Sub>3< / com:Sub>) <com:Math><semantics>δ<annotation encoding="application / x-tex">\delta< / annotation>< / semantics>< / com:Math> 8.39 (s, 1H), 7.57 (d, <com:Math><semantics>J=8.1<annotation encoding="application / x-tex">J = 8.1< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.51 (t, <com:Math><semantics>J=8.0<annotation encoding="application / x-tex">J = 8.0< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.39 (d, <com:Math><semantics>J=6.9 Hz<annotation encoding="application / x-tex">J = 6.9 \text{ Hz}< / annotation>< / semantics>< / com:Math>, 1H), 7.13 (dd, <com:Math><semantics>J=11.7<annotation encoding="application / x-tex">J = 11.7< / annotation>< / semantics>< / com:Math>, 4.7 Hz, 1H), 6.78 (t, <com:Math><semantics>J=7.4 Hz<annotation encoding="application / x-tex">J = 7.4 \text{ Hz}< / annotation>< / semantics>< / com:Math>, 1H), 6.48 (q, <com:Math><semantics>J=7.4 Hz<annotation encoding="application / x-tex">J = 7.4 \text{ Hz}< / annotation>< / semantics>< / com:Math>) 6.9 Hz, 1H), 5.36 (brs, 2H), 4.18 (td, <com:Math><semantics>J=11.2<annotation encoding="application / x-tex">J = 11.2< / annotation>< / semantics>< / com:Math>, 3.1 Hz, 1H), 4.05 (dd, <com:Math><semantics>J=14.4<annotation encoding="application / x-tex">J = 14.4< / annotation>< / semantics>< / com:Math>, 7.3 Hz, 1H), <com:Math><semantics>4.03−3.96<annotation encoding="application / x-tex">4.03 - 3.96< / annotation>< / semantics>< / com:Math> (m, 1H), <com:Math><semantics>3.87<annotation encoding="application / x-tex">3.87< / annotation>< / semantics>< / com:Math> (s, 3H), <com:Math><semantics>3.80<annotation encoding="application / x-tex">3.80< / annotation>< / semantics>< / com:Math> (d, <com:Math><semantics>J=10.3<annotation encoding="application / x-tex">J = 10.3< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>3.65<annotation encoding="application / x-tex">3.65< / annotation>< / semantics>< / com:Math> (dd, <com:Math><semantics>J=11.4<annotation encoding="application / x-tex">J = 11.4< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>9.2<annotation encoding="application / x-tex">9.2< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>3.37(d,J=10.8Hz,1H),2.77(d,J=10.6Hz,1H),2.40(dd,J=11.4,9.2Hz,1H),1.87(d,J=10.8Hz,1H),2.77(d,J=10.6Hz,1H),2.40(dd,J=11.4,9.2Hz,1H),1.87(d,J=10.8Hz,1H),2.77(d,J=10.6H<annotation encoding="application / x-tex">3.37 (d, J = 10.8 Hz, 1H), 2.77 (d, J = 10.6 Hz, 1H), 2.40 (dd, J = 11.4, 9.2 Hz, 1H), 1.87 (d, J = 10.8 Hz, 1H), 2.77 (d, J = 10.6 Hz, 1H), 2.40 (dd, J = 11.4, 9.2 Hz, 1H), 1.87 (d, J = 10.8 Hz, 1H), 2.77 (d, J = 10.6 H< / annotation>< / semantics>< / com:Math> <com:Math><semantics>J=6.9 Hz,3H<annotation encoding="application / x-tex">J = 6.9 \text{ Hz}, 3\text{H}< / annotation>< / semantics>< / com:Math>). LC-MS (ESI-MS): 569 [M+H]<com:Sup>+< / com:Sup>. Embodiment 12: Synthesis of Target Compound II-C-2 [Image disponible dans le document PDF, Image available in the PDF document]

[0299] Step 1: 3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (1.0mmol) was placed in a 20ml reaction flask with with 5ml of DMF, and stirred to dissolve at room temperature, then mixed in turn with 4-(2-bromoethyl)-5-chloro-4-oxoquinazolin- 3(4H)-yl)piperidine-1-carboxylic acid, tert-butyl ester (1.1mmol) and potassium carbonate (2.0mmol), and heated to 40°C to react for about 15h. When the reaction was completed as detected by the TLC test, ethyl acetate was added to extract, and washed with water. The ethyl acetate layer was separated, dried over anhydrous sodium sulphate and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 0.46 g of Intermediate 13 with a yield of 69%.

[0300] Step 2: Intermediate 13 (0.6mmol) was placed in a 20ml reaction flask with 6ml of dichloromethane, then mixed with 1ml of trifluoroacetic acid to react for about 12h at room temperature. When the reaction was basically completed as detected by the TLC test, water and dichloromethane were added to extract. The organic phase was washed with saturated aqueous sodium bicarbonate solution. The dichloromethane layer was separated, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 0.27g of the target product, Compound II-C-2, with a yield of 80%. LC-MS (ESI-MS): 567 [M+H]<com:Sup>+< / com:Sup>. Embodiment 13: Synthesis of Target Compound II-C-3 [Image disponible dans le document PDF, Image available in the PDF document]

[0301] Compound II-C-2 (0.3mmol) was placed in a 20ml reaction flask with 6ml of dichloromethane, and then mixed with acetic acid (0.3mmol) and 1-ethyl-(3- dimethylaminopropyl)carbodiimide hydrochloride (0.6mmol) to react for about 3h at room temperature. When the reaction was completed as detected by the TLC test, water and dichloromethane were added to extract. The organic phase was separated, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 151mg of a white product, compound II-C-3, with a yield of 83%. LC-MS (ESI-MS): 609 [M+H]<com:Sup>+< / com:Sup>. Embodiment 14: Synthesis of Target Compound II-C-4 [Image disponible dans le document PDF, Image available in the PDF document]

[0302] Compound II-C-2 (0.3mmol) was placed in a 20ml reaction flask with 4ml of tetrahydrofuran, and then mixed with trifluoromethyl iodoethane (0.3mmol) and potassium carbonate (0.6mmol) to react for about 2h at room temperature as the reaction was completed as detected by the TLC test, then quenched with water and extracted with ethyl acetate. The organic phase was dried with anhydrous sodium sulfate and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 101mg of an off-white solid product, Compound II-C-4, with a yield of 52%. LC-MS (ESI- MS): <com:Math><semantics>649[M+H]+<annotation encoding="application / x-tex">649 [M+H]^+< / annotation>< / semantics>< / com:Math>. Embodiment 15: Synthesis of Target Compound II-C-7 [Image disponible dans le document PDF, Image available in the PDF document]

[0303] Step 1: 3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (9a, 1.0mmol) was placed in a 20ml reaction flask with 5ml of DMF, and stirred to dissolve at room temperature, then mixed in turn with 4-(2-bromoethyl)-5-chloro-4-oxoquinazolin- 3(4H)-yl)piperazine-1-carboxylic acid, tert-butyl ester (4p, 1.1mmol) and potassium carbonate (2.0mmol), and heated to 40°C to react for about 15h. When the reaction was completed as detected by the TLC test, ethyl acetate was added to extract, and washed with water. The ethyl acetate layer was separated, dried over anhydrous sodium sulphate and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 0.42 g of Intermediate 14 with a yield of 63%.

[0304] Step 2: Intermediate 14 (0.6mmol) was placed in a 20ml reaction flask with 6ml of dichloromethane, then mixed with 1ml of trifluoroacetic acid to react for about 12h at room temperature. When the reaction was basically completed as detected by the TLC test, water and dichloromethane were added to extract. The organic phase was washed with saturated aqueous sodium bicarbonate solution. The dichloromethane layer was separated, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 0.28g of the target product, Compound II-C-7, with a yield of 82%. <com:Sup>1< / com:Sup>H NMR (500 MHz, DMSO) <com:Math><semantics>δ<annotation encoding="application / x-tex">\delta< / annotation>< / semantics>< / com:Math> 9.11 (s, 3H), 8.32 (s, 1H), 7.78 <com:Math><semantics>(t,J=7.9 Hz,1H),7.67(d,J=8.1 Hz,1H),7.59(d,J=8.0 Hz,1H),7.20(t,J=7.9 Hz,1H),<annotation encoding="application / x-tex">(t, J = 7.9 \text{ Hz}, 1\text{H}), 7.67 (d, J = 8.1 \text{ Hz}, 1\text{H}), 7.59 (d, J = 8.0 \text{ Hz}, 1\text{H}), 7.20 (t, J = 7.9 \text{ Hz}, 1\text{H}),< / annotation>< / semantics>< / com:Math> 7.05 (t, <com:Math><semantics>J=8.1 Hz<annotation encoding="application / x-tex">J = 8.1 \text{ Hz}< / annotation>< / semantics>< / com:Math>, 1H), 6.44 (dd, <com:Math><semantics>J=13.3<annotation encoding="application / x-tex">J = 13.3< / annotation>< / semantics>< / com:Math>, 6.8 Hz, 1H), 4.04–3.89 (m, 2H), 3.88 (s, 3H), 3.45 <com:Math><semantics>(d,J=11.7 Hz,2H),3.02(t,J=11.5 Hz,1H),2.85(d,J=11.3 Hz,1H),1.89(d,J=11.1 Hz,1Hz)<annotation encoding="application / x-tex">(d, J = 11.7 \text{ Hz}, 2\text{H}), 3.02 (t, J = 11.5 \text{ Hz}, 1\text{H}), 2.85 (d, J = 11.3 \text{ Hz}, 1\text{H}), 1.89 (d, J = 11.1 \text{ Hz}, 1\text{Hz})< / annotation>< / semantics>< / com:Math> 1H), 1.72 (d, <com:Math><semantics>J−6.5<annotation encoding="application / x-tex">J - 6.5< / annotation>< / semantics>< / com:Math> Hz, 3H), 1.42 (t, <com:Math><semantics>J−10.8<annotation encoding="application / x-tex">J - 10.8< / annotation>< / semantics>< / com:Math> Hz, 1H). LC-MS (ESI-MS): 567 [M+H]<com:Sup>+< / com:Sup>. Embodiment 16: Synthesis of Target Compound II-C-8 [Image disponible dans le document PDF, Image available in the PDF document]

[0305] Step 1: 3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (9a, 1.0mmol) was placed in a 20ml reaction flask with 5ml of DMF, and stirred to dissolve at room temperature, then mixed in turn with 4-(2-bromoethyl)-5-chloro-4-oxoquinazolin- 3(4H)-yl)-2,2-dimethylpiperazine-1-carboxylic acid, tert-butyl ester (4r, 1.1mmol) and potassium carbonate (2.0mmol), and heated to 40°C to react for about 15h. When the reaction was completed as detected by the TLC test, ethyl acetate was added to extract, and washed with water. The ethyl acetate layer was separated, dried over anhydrous sodium sulphate and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 0.38 g of Intermediate 15 with a yield of 55%.

[0306] Step 2: Intermediate 15 (0.6mmol) was placed in a 20ml reaction flask with 6ml of dichloromethane, and then mixed with 1ml of trifluoroacetic acid to react for about 12h at room temperature. When the reaction was basically completed as detected by the TLC test, water and dichloromethane were added to extract, and the organic phase was washed with saturated aqueous sodium bicarbonate solution. The dichloromethane layer was separated, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 0.28g of the target product, Compound II-C-8, with a yield of 79%. LC-MS (ESI-MS): 596 [M+H]<com:Sup>+< / com:Sup>. Embodiment 17: Synthesis of Target Compound II-C-9 [Image disponible dans le document PDF, Image available in the PDF document]

[0307] Step 1: 3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (9a, 1.0mmol) was placed in a 20ml reaction flask with 5ml of DMF, and stirred to dissolve at room temperature, then mixed in turn with 4-(2-bromoethyl)-5-chloro-4-oxoquinazolin- 3(4H)-yl)-2-(trifluoromethyl)piperazine-1-carboxylic acid, tert-butyl ester (4q, 1.1mmol) and potassium carbonate (2.0mmol), and heated to 40°C to react for about 15h. When the reaction was complete as detected by the TLC test, ethyl acetate was added to extracted, and washed with water. The ethyl acetate layer was separated, dried with anhydrous sodium sulphate and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 0.36 g of Intermediate 16, with a yield of 49%.

[0308] Ste 2: Intermediate 16 (0.6mmol) was placed in a 20ml reaction flask with 6ml of dichloromethane, and then mixed with 1ml of trifluoroacetic acid to react for about 12h at room temperature. When the reaction was basically completed as detected by the TLC test, water and dichloromethane were added to extract. The organic phase was washed with saturated aqueous sodium bicarbonate solution. The dichloromethane layer was separated, dried with anhydrous sodium sulfate and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 0.28g of the target product, Compound II-C-9, with a yield of 75%. LC-MS (ESI-MS): 636 [M+H]<com:Sup>+< / com:Sup>. Embodiment 18: Synthesis of Target Compound II-C-10 [Image disponible dans le document PDF, Image available in the PDF document]

[0309] Compound II-C-7 (0.5mmol) was placed in a 20ml reaction flask with 5ml of tetrahydrofuran, and then mixed with trifluoromethyl iodoethane (0.5mmol) and potassium carbonate (1.0mmol) to react for about 2h at room temperature and the reaction was completed as detected by the TLC test, then quench with water and extract with ethyl acetate The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 172mg of off- white solid, compound II-C-10, with a yield of 53%. LC-MS (ESI-MS): 650 [M+H]<com:Sup>+< / com:Sup>. Embodiment 19: synthesis of target compound II-C-11 [Image disponible dans le document PDF, Image available in the PDF document]

[0310] Place compound II-C-7 (0.5mmol) and 5ml of tetrahydrofuran, mix with iodomethane (0.45mmol) and potassium carbonate (1.0mmol) in a 20ml reaction flask, react for about 2h at room temperature, quench with water and extract with ethyl acetate, dry the organic phase with anhydrous sodium sulfate and concentrate under reduced pressure, purify the resulting residue by silica gel column chromatography to obtain 125mg of white-like solid, compound II-C-11 with a yield of 43%. <com:Sup>1< / com:Sup>H NMR (500 MHz, CDCl<com:Sub>3< / com:Sub>) δ 8.44 (s, 1H), 7.60 (d, <com:Math><semantics>J=7.9 Hz<annotation encoding="application / x-tex">J = 7.9 \text{ Hz}< / annotation>< / semantics>< / com:Math>, 1H), 7.55 (t, <com:Math><semantics>J=7.8 Hz<annotation encoding="application / x-tex">J = 7.8 \text{ Hz}< / annotation>< / semantics>< / com:Math>, 1H), 7.43 (d, <com:Math><semantics>J=7.5 Hz<annotation encoding="application / x-tex">J = 7.5 \text{ Hz}< / annotation>< / semantics>< / com:Math>, 1H), 7.19 (t, <com:Math><semantics>J=7.6 Hz<annotation encoding="application / x-tex">J = 7.6 \text{ Hz}< / annotation>< / semantics>< / com:Math>, 1H), 6.84 (t, <com:Math><semantics>J=7.8<annotation encoding="application / x-tex">J = 7.8< / annotation>< / semantics>< / com:Math> Hz, 1H), 6.50 (dd, <com:Math><semantics>J=13.2<annotation encoding="application / x-tex">J = 13.2< / annotation>< / semantics>< / com:Math>, 6.6 Hz, 1H), 5.51 (brs, 2H), 4.23 (t, <com:Math><semantics>J=10.4<annotation encoding="application / x-tex">J = 10.4< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>4.06<annotation encoding="application / x-tex">4.06< / annotation>< / semantics>< / com:Math> (t, <com:Math><semantics>J=10.1<annotation encoding="application / x-tex">J = 10.1< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>3.94<annotation encoding="application / x-tex">3.94< / annotation>< / semantics>< / com:Math> (s, 3H), <com:Math><semantics>2.84<annotation encoding="application / x-tex">2.84< / annotation>< / semantics>< / com:Math> (dd, <com:Math><semantics>J=33.7<annotation encoding="application / x-tex">J = 33.7< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>10.2<annotation encoding="application / x-tex">10.2< / annotation>< / semantics>< / com:Math> Hz, 2H), <com:Math><semantics>2.40<annotation encoding="application / x-tex">2.40< / annotation>< / semantics>< / com:Math> (d, <com:Math><semantics>J=11.5<annotation encoding="application / x-tex">J = 11.5< / annotation>< / semantics>< / com:Math> Hz, 2H), 2.17 (s, 3H), 2.08 (d, <com:Math><semantics>J=9.6<annotation encoding="application / x-tex">J = 9.6< / annotation>< / semantics>< / com:Math> Hz, 2H), 1.94 (d, <com:Math><semantics>J=6.6<annotation encoding="application / x-tex">J = 6.6< / annotation>< / semantics>< / com:Math> Hz, 3H). LC-MS (ESI-MS): 582 [M+H]<com:Sup>+< / com:Sup>. Embodiment 20: synthesis of target compound II-C-12 [Image disponible dans le document PDF, Image available in the PDF document]

[0311] Place compound II-C-7 (0.3mmol) and 6ml of dichloromethane in a 20ml reaction flask, mix with acetic acid (0.3mmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (0.6mmol), react for about 3h at room temperature, complete the TLC test, extract with water and dichloromethane, separate the organic phase to dry with anhydrous sodium sulfate and concentrate under reduced pressure, purify the resulting residue by silica gel column chromatography to obtain 155mg of white product, compound II-C-12 with a yield of 85%. LC-MS (ESI-MS): 610 [M+H]<com:Sup>+< / com:Sup>. Embodiment 21: synthesis of compound II-C-13 [Image disponible dans le document PDF, Image available in the PDF document]

[0312] place 3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (9a, 1.0mmol) and 5ml of DMF in a 20ml reaction flask, stir to dissolve at room temperature, then add in turn 2-(1-bromoethyl)-3-(3-methoxypropyl)quinazolin-4(3H)-one (4j, 1.1mmol) and potassium carbonate (2.0mmol), heat to 40°C to react for about 15h, complete TLC test, extract with ethyl acetate, wash with water, separate the ethyl acetate layer, dry with anhydrous sodium sulphate and concentrate under reduced pressure, purify the resulting residue through silica gel column chromatography to obtain 0.35g of solid product, compound II-C-13 with a yield of 67%. LC-MS (ESI-MS): 522 [M+H]<com:Sup>+< / com:Sup>. Embodiment 22: synthesis of compound II-C-14 [Image disponible dans le document PDF, Image available in the PDF document]

[0313] place 3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (9a, 1.0mmol) and 5ml of DMF in a 20ml reaction flask, stir to dissolve at room temperature, then add in turn 2-(1-bromoethyl)-5-chloro-3-(3-methoxypropyl)quinazolin-4(3H)-one (4k, 1.1mmol) and potassium carbonate (2.0mmol), heat to 40°C to react for about 15h, complete TLC test, extract with ethyl acetate, wash with water, separate the ethyl acetate layer, dry with anhydrous sodium sulphate and concentrate under reduced pressure, purify the resulting residue through silica gel column chromatography to obtain 0.36g of solid product, compound II-C-14 with a yield of 65%. <com:Sup>1< / com:Sup>H NMR (500 MHz, DMSO) <com:Math><semantics>δ<annotation encoding="application / x-tex">\delta< / annotation>< / semantics>< / com:Math> 8.30 (s, 1H), 7.72 (t, <com:Math><semantics>J=7.5<annotation encoding="application / x-tex">J = 7.5< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.57 (dd, <com:Math><semantics>J=28.8<annotation encoding="application / x-tex">J = 28.8< / annotation>< / semantics>< / com:Math>, 7.5 Hz, 2H), 7.18 (s, 1H), 7.08 (d, <com:Math><semantics>J=7.2<annotation encoding="application / x-tex">J = 7.2< / annotation>< / semantics>< / com:Math> Hz, 1H), 6.40 (d, <com:Math><semantics>J=5.7<annotation encoding="application / x-tex">J = 5.7< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>4.06−3.75<annotation encoding="application / x-tex">4.06 - 3.75< / annotation>< / semantics>< / com:Math> (m, 2H), <com:Math><semantics>3.84<annotation encoding="application / x-tex">3.84< / annotation>< / semantics>< / com:Math> (s, 3H), <com:Math><semantics>3.05<annotation encoding="application / x-tex">3.05< / annotation>< / semantics>< / com:Math> (s, 3H), <com:Math><semantics>2.96<annotation encoding="application / x-tex">2.96< / annotation>< / semantics>< / com:Math> (brs, 2H), <com:Math><semantics>1.85−1.76<annotation encoding="application / x-tex">1.85 - 1.76< / annotation>< / semantics>< / com:Math> (m, 5H). LC-MS (ESI-MS): 556 [M+H]<com:Sup>+< / com:Sup>. Embodiment 23: synthesis of compound II-C-15 [Image disponible dans le document PDF, Image available in the PDF document]

[0314] place 3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (9a, 1.0mmol) and 5ml of DMF in a 20ml reaction flask, stir to dissolve at room temperature, then add in turn the 2-(1-bromoethyl)-5-chlorine-3-(2-(tetrahydrofuran-2-yl)ethyl)quinazolin- 4(3II)-one (4l, 1.1mmol) and potassium carbonate (2.0mmol), heat to 40°C to react for about 15h, complete TLC test, extract with ethyl acetate, wash with water, separate the ethyl acetate layer, dry with anhydrous sodium sulphate and concentrate under reduced pressure, purify the resulting residue through silica gel column chromatography to obtain 0.30g of solid product, compound II-C-15 with a yield of 52%. LC-MS (ESI-MS): 582 [M+H]<com:Sup>+< / com:Sup>. Embodiment 24: Synthesis of Target Compounds II-A-1a and II-A-1b [Image disponible dans le document PDF, Image available in the PDF document]

[0315] Place intermediate 9a (10mmol), (R)-12a or (S)-12a (20mmol), triphenylphosphine (5.3g, 20mmol) and tetrahydrofuran 50ml in a reaction flask, cool to 0°C, drip diisopropyl azodicarboxylate (3.9ml, 20mmol); heat to room temperature after dripping to react for about 6h so TLC test is completed, concentrate under reduced pressure to remove most of the tetrahydrofuran, and purify the residue by silica gel column chromatography to obtain a white solid product.

[0316] Target compound II-A-1a: 3.1g, 58% yield; Chiral assay ee%: 97.8%, chiral-HPLC being performed using a normal-phase CHIRALCEL OD-H column with a retention time of 13.86min. LC-MS (ESI-MS): 526 [M+H]<com:Sup>+< / com:Sup>.

[0317] Target compound II-A-1b: 2.9g, 55% yield; Chiral assay ee%: 98.2%, chiral-HPLC being performed using a normal-phase CHIRALCEL OD-H column with a retention time of 17.24min. LC-MS (ESI-MS): 526 [M+H]<com:Sup>+< / com:Sup>. Embodiment 25: synthesis of target compounds <com:Math><semantics>Π<annotation encoding="application / x-tex">\Pi< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>Λ<annotation encoding="application / x-tex">\Lambda< / annotation>< / semantics>< / com:Math>-2a and <com:Math><semantics>Π<annotation encoding="application / x-tex">\Pi< / annotation>< / semantics>< / com:Math>-<com:Math><semantics>Λ<annotation encoding="application / x-tex">\Lambda< / annotation>< / semantics>< / com:Math>-2b [Image disponible dans le document PDF, Image available in the PDF document]

[0318] Place intermediate 9b (10mmol), (R)-12a or (S)-12a (20mmol), triphenylphosphine (5.3g, 20mmol) and tetrahydrofuran 50ml in a reaction flask, cool to 0°C, drip diisopropyl azodicarboxylate (3.9ml, 20mmol); heat to room temperature after dripping to react for about 6h so TLC test is completed, concentrate under reduced pressure to remove most of the tetrahydrofuran, and purify the residue by silica gel column chromatography to obtain a white solid product.

[0319] Target compound II-A-2a: 3.1g, 56% yield; Chiral assay ee%: 98.2%, chiral-HPLC being performed using a normal-phase CHIRALCEL OD-H column with a retention time of 14.10min. LC-MS (ESI-MS): 554 [M+H]<com:Sup>+< / com:Sup>.

[0320] Target compound II-A-2b: 3.0g, 54% yield; Chiral assay ee%: 97.8%, chiral-HPLC being performed using a normal-phase CHIRALCEL OD-H column with a retention time of 16.90min. LC-MS (ESI-MS): 554 [M+H]<com:Sup>+< / com:Sup>. Embodiment 26: Synthesis of Target Compounds II-A-7a and II-A-7b [Image disponible dans le document PDF, Image available in the PDF document]

[0321] Place intermediate 9a (10mmol), (R)-12b or <com:Math><semantics>(S)−12b<annotation encoding="application / x-tex">(S)-12b< / annotation>< / semantics>< / com:Math> (20mmol), triphenylphosphine (5.3g, 20mmol) and tetrahydrofuran 50ml in a reaction flask, cool to 0°C, drip diisopropyl azodicarboxylate (3.9ml, 20mmol); heat to room temperature after dripping to react for about 6h so TLC test is completed, concentrate under reduced pressure to remove most of the tetrahydrofuran, and purify the residue by silica gel column chromatography to obtain a white solid product.

[0322] Target compound II-A-7a: 3.1g, 55% yield; Chiral assay ee%: 98.6%, chiral-HPLC being performed using a normal-phase CHIRALCEL OD-H column with a retention time of 12.09min. LC-MS (ESI-MS): 560 [M+H]<com:Sup>+< / com:Sup>.

[0323] Target compound II-A-7b: 3.0g, 54% yield; Chiral assay ee%: 98.1%, chiral-HPLC being performed using a normal-phase CHIRALCEL OD-H column with a retention time of 19.90min. LC-MS (ESI-MS): 560 [M+H]<com:Sup>+< / com:Sup>. Embodiment 27: Synthesis of Target Compounds II-A-8a and II-A-8b [Image disponible dans le document PDF, Image available in the PDF document]

[0324] Intermediate 9a (10mmol), (R)-12c or (S)-12c (20mmol) and triphenylphosphine (5.3g, 20mmol) were placed in a reaction flask with 50ml of tetrahydrofuran, cooled to 0°C, dripped with diisopropyl azodicarboxylate (3.9ml, 20mmol) and then heated to room temperature after dripping to react for about 6h till the reaction was completed as detected by the TLC test. Most of the tetrahydrofuran was then removed by concentration under reduced pressure, and the residue was purified by silica gel column chromatography to obtain a white solid product.

[0325] Target compound II-A-8a: 3.1g, 57% yield; chiral assay ee%: 98.7%, chiral-HPLC being performed using a normal-phase CHIRALCEL OD-H column with a retention time of 12.75min. LC-MS (ESI-MS): 544 [M+H]<com:Sup>+< / com:Sup>.

[0326] Target compound II-A-8b: 3.0g, 55% yield; chiral assay ee%: 98.2%, chiral-HPLC being performed using a normal-phase CHIRALCEL OD-H column with a retention time of 18.99min. LC-MS (ESI-MS): 544 [M+H]<com:Sup>+< / com:Sup>. Embodiment 28: Synthesis of Target Compounds II-B-2a and II-B-2b [Image disponible dans le document PDF, Image available in the PDF document]

[0327] Intermediate 9a (10mmol), (R)-12d or (S)-12d (20mmol) and triphenylphosphine (5.3g, 20mmol) were placed in a reaction flask with 50ml of tetrahydrofuran, cooled to 0°C, drippd with diisopropyl azodicarboxylate (3.9ml, 20mmol) and then heated to room temperature after dripping to react for about 6h till the reaction was completed as detected by the TLC test. Most of the tetrahydrofuran was removed by concentration under reduced pressure, and the residue was purified by silica gel column chromatography to obtain a white solid product.

[0328] Target compound II-B-2a: 2.7g, 48% yield; chiral assay ee%: 98.0%, chiral-HPLC being performed using a normal-phase CHIRALCEL OD-H column with a retention time of 13.42min. LC-MS (ESI-MS): 561 [M+H]<com:Sup>+< / com:Sup>.

[0329] Target compound II-B-2b: 2.6g, 46% yield; chiral assay ee%: 97.8%, chiral-HPLC being performed using a normal-phase CHIRALCEL OD-H column with a retention time of 23.15min. LC-MS (ESI-MS): 561 [M+H]<com:Sup>+< / com:Sup>. Embodiment 29: Synthesis of Compounds II-D-1~II-D-4

[0330] Compound II-A-1a (231mg, 0.4 mmol) was placed in a flask, mixed with 6ml of isopropanol, stirred, mixed with an inorganic or organic acid (0.44mmol), heated to reflux, reacted for 1h with stirring, slowly cooled to room temperature, desolvated and filtered. The solid was collected, dried in vacuum at room temperature to obtain the solid product.

[0331] Compound II-D-1: the acid added was hydrochloric acid (0.44mmol). The final product II-D-1 was a white solid, 241mg with 98.1% yield. LC-MS (ESI-MS): 526 [M+H]<com:Sup>+< / com:Sup>.

[0332] Compound II-D-2: the acid added was sulfuric acid (0.44mmol). The final product II-D-2 was a white solid, 250mg with 99.5% yield. LC-MS (ESI-MS): 526 [M+H]<com:Sup>+< / com:Sup>.

[0333] Compound II-D-3: the acid added was toluenesulfonic acid monohydrate (84mg, 0.44mmol). The final product II-D-3 was a white solid, 273mg with 91% yield. <com:Sup>1< / com:Sup>H NMR (400 MHz, DMSO) <com:Math><semantics>δ<annotation encoding="application / x-tex">\delta< / annotation>< / semantics>< / com:Math> 8.17 (s, 1H), 7.84 (t, <com:Math><semantics>J=8.0<annotation encoding="application / x-tex">J = 8.0< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.76 (dd, <com:Math><semantics>J=8.2<annotation encoding="application / x-tex">J = 8.2< / annotation>< / semantics>< / com:Math>, 1.1 Hz, 1H), 7.65 (dd, <com:Math><semantics>J=12.9,5.1 Hz,2H,7.50−7.43 (m, 3H),7.37−7.25 (m, 1H),7.22−7.04 (m, 4H),6.93 (dd, 1H)<annotation encoding="application / x-tex">J = 12.9, 5.1 \text{ Hz}, 2\text{H}, 7.50-7.43 \text{ (m, 3H)}, 7.37-7.25 \text{ (m, 1H)}, 7.22-7.04 \text{ (m, 4H)}, 6.93 \text{ (dd, 1H)}< / annotation>< / semantics>< / com:Math> <com:Math><semantics>J=11.1,4.3 Hz,1H,6.44 (d,J=8.2 Hz,1H),5.99 (q,J=6.6 Hz,1H),3.93 (s,3H),2.29 (s,3.20 Hz,1.20 Hz)<annotation encoding="application / x-tex">J = 11.1, 4.3 \text{ Hz}, 1\text{H}, 6.44 \text{ (d}, J = 8.2 \text{ Hz}, 1\text{H}), 5.99 \text{ (q}, J = 6.6 \text{ Hz}, 1\text{H}), 3.93 \text{ (s}, 3\text{H}), 2.29 \text{ (s}, 3.20 \text{ Hz}, 1.20 \text{ Hz})< / annotation>< / semantics>< / com:Math> 3H), 1.78 (d, <com:Math><semantics>J=6.7 Hz<annotation encoding="application / x-tex">J = 6.7 \text{ Hz}< / annotation>< / semantics>< / com:Math>, 3H), LC-MS (ESI-MS): 526[M+H]<com:Sup>+< / com:Sup>.

[0334] Compound II-D-4: the acid added was methanesulfonic acid (29µl, 0.44mmol). The final product II-D-4 was a white solid, 260mg with 96.5% yield. LC-MS (ESI-MS): 526 <com:Math><semantics>[M+H]+<annotation encoding="application / x-tex">[M+H]^{+}< / annotation>< / semantics>< / com:Math>. Embodiment 30: Synthesis of Compounds II-D-5~II-D-17

[0335] Compound II-A-7a (0.4 mmol) was placed in a flask, mixed 6ml of isopropanol, stirred, mixed with an inorganic or organic acid (0.44mmol), heated to reflux, reacted for 1h with stirring, slowly cooled to room temperature, desolvated and filtered. The solid was collected and dried in vacuum at room temperature to obtain the solid product.

[0336] Compound II-D-5: the acid added was hydrochloric acid (0.44mmol). The final product II-D-5 was a white solid, 233mg with 99.5% yield. <com:Sup>1< / com:Sup>H NMR (500 MHz, DMSO) δ 8.04 (s, 1H), 7.84 (t, <com:Math><semantics>J=8.0<annotation encoding="application / x-tex">J = 8.0< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.75 (d, <com:Math><semantics>J=8.2<annotation encoding="application / x-tex">J = 8.2< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.64 (d, <com:Math><semantics>J=7.9<annotation encoding="application / x-tex">J = 7.9< / annotation>< / semantics>< / com:Math> Hz, 2H), 7.46 <Formule mathématique disponible dans le document PDF, Math available in the PDF document>(t, J = 7.7 \text{ Hz}, 1\text{H}), 7.29 (t, J = 7.7 \text{ Hz}, 1\text{H}), 7.20 - 7.05 (m, 2\text{H}), 6.87 (t, J = 7.7 \text{ Hz}, 1\text{H}), 6.37 (m, 2\text{H}), 6.87 (t, J = 7.7 \text{ Hz}, 1\text{H}), 6.37 (t, J = 7.7 \text{ Hz}, 1\text{H}), 6.37 (t, J = 7.7 \text{ <com:Math><semantics>(d,J=8.0 Hz,1H),5.95(q,J=6.6 Hz,1H),3.92(s,3H),1.75(d,J=6.6 Hz,3H).<annotation encoding="application / x-tex">(d, J = 8.0 \text{ Hz}, 1\text{H}), 5.95 (q, J = 6.6 \text{ Hz}, 1\text{H}), 3.92 (s, 3\text{H}), 1.75 (d, J = 6.6 \text{ Hz}, 3\text{H}).< / annotation>< / semantics>< / com:Math> LC-MS (ESI-MS): 560 [M+H]<com:Sup>+< / com:Sup>.

[0337] Compound II-D-6: the acid added was sulfuric acid (0.44mmol). The final product II-D-6 was an off-white solid, 244mg with 99.5% yield. <com:Sup>1< / com:Sup>H NMR (500 MHz, DMSO) δ 8.06 (s, 1H), 7.84 (t, <com:Math><semantics>J=8.0<annotation encoding="application / x-tex">J = 8.0< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.75 (d, <com:Math><semantics>J=8.1<annotation encoding="application / x-tex">J = 8.1< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.64 (d, <com:Math><semantics>J=7.8<annotation encoding="application / x-tex">J = 7.8< / annotation>< / semantics>< / com:Math> Hz, 2H), 7.46 (t, <com:Math><semantics>J=8.0<annotation encoding="application / x-tex">J = 8.0< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.85 (t, <com:Math><semantics>J=8.0<annotation encoding="application / x-tex">J = 8.0< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.86 (t, <com:Math><semantics>J=8.0<annotation encoding="application / x-tex">J = 8.0< / annotation>< / semantics>< / com:Math> Hz, 2H), 7.86 (t, <com:Math><semantics>J=8.0<annotation encoding="application / x-tex">J = 8.0< / annotation>< / semantics>< / com:Math> Hz, 2H), 7.86 (t, <com:Math><semantics>J=8.<annotation encoding="application / x-tex">J = 8.< / annotation>< / semantics>< / com:Math> <com:Math><semantics>=7.7 Hz,1H,7.30 (t, J=7.5 Hz,1H),7.22−7.06 (m, 2H),6.88 (t, J=7.7 Hz,1H),6.38 (d, 2H)<annotation encoding="application / x-tex">= 7.7 \text{ Hz}, 1\text{H}, 7.30 \text{ (t, } J = 7.5 \text{ Hz}, 1\text{H}), 7.22 - 7.06 \text{ (m, 2H)}, 6.88 \text{ (t, } J = 7.7 \text{ Hz}, 1\text{H}), 6.38 \text{ (d, 2H)}< / annotation>< / semantics>< / com:Math> <com:Math><semantics>J=8.1 Hz<annotation encoding="application / x-tex">J = 8.1 \text{ Hz}< / annotation>< / semantics>< / com:Math>, 1H), 5.96 (q, <com:Math><semantics>J=6.6 Hz<annotation encoding="application / x-tex">J = 6.6 \text{ Hz}< / annotation>< / semantics>< / com:Math>, 1H), 3.92 (s, 3H), 1.75 (d, <com:Math><semantics>J=6.6 Hz<annotation encoding="application / x-tex">J = 6.6 \text{ Hz}< / annotation>< / semantics>< / com:Math>, 3H). LC-MS (ESI- MS): <com:Math><semantics>560[M+H]+<annotation encoding="application / x-tex">560 [M+H]^+< / annotation>< / semantics>< / com:Math>.

[0338] Compound II-D-7: the acid added was phosphoric acid (0.44mmol). The final product II-D-7 was an off-white solid, 207mg with 78.6% yield. <com:Sup>1< / com:Sup>H NMR (500 MHz, DMSO) <com:Math><semantics>δ<annotation encoding="application / x-tex">\delta< / annotation>< / semantics>< / com:Math> 7.88 (s, 1H), 7.83 (t, <com:Math><semantics>J=8.0<annotation encoding="application / x-tex">J = 8.0< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.75 (d, <com:Math><semantics>J=8.1<annotation encoding="application / x-tex">J = 8.1< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.61 (dd, <com:Math><semantics>J=13.8<annotation encoding="application / x-tex">J = 13.8< / annotation>< / semantics>< / com:Math>, 7.9 Hz, 2H), 7.43 (t, <com:Math><semantics>J=7.6<annotation encoding="application / x-tex">J = 7.6< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.24 (t, <com:Math><semantics>J=7.6<annotation encoding="application / x-tex">J = 7.6< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.10 (dd, <com:Math><semantics>J=12.8<annotation encoding="application / x-tex">J = 12.8< / annotation>< / semantics>< / com:Math>, 7.5 Hz, 2H), 6.81 (t, <com:Math><semantics>J=7.7 Hz<annotation encoding="application / x-tex">J = 7.7 \text{ Hz}< / annotation>< / semantics>< / com:Math>, 1H), 6.31 (d, <com:Math><semantics>J=7.8 Hz<annotation encoding="application / x-tex">J = 7.8 \text{ Hz}< / annotation>< / semantics>< / com:Math>, 1H), 5.90 (q, <com:Math><semantics>J=6.6 Hz<annotation encoding="application / x-tex">J = 6.6 \text{ Hz}< / annotation>< / semantics>< / com:Math>, 1H), 3.91 (s, 3H), 1.72 (d, <com:Math><semantics>J=<annotation encoding="application / x-tex">J =< / annotation>< / semantics>< / com:Math> 6.6 Hz, 3H). LC-MS (ESI-MS): 560 [M+H]<com:Sup>+< / com:Sup>.

[0339] Compound II-D-8: the acid added was toluenesulfonic acid monohydrate (84mg, 0.44mmol). The final product II-D-8 was an off-white solid, 268mg with 91.5% yield. <com:Sup>1< / com:Sup>H NMR (400 MHz, DMSO) <com:Math><semantics>δ<annotation encoding="application / x-tex">\delta< / annotation>< / semantics>< / com:Math> 8.20 – 8.05 (m, 2H), 7.97–7.87 (m, 1H), 7.82 (d, <com:Math><semantics>J<annotation encoding="application / x-tex">J< / annotation>< / semantics>< / com:Math> = 7.9 Hz, 1H), 7.68 (d, <com:Math><semantics>J=8.0<annotation encoding="application / x-tex">J = 8.0< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.65–7.56 (m, 1H), 7.54–7.39 (m, 3H), 7.34–7.24 (m, 1H), 7.20– 7.01 (m, 3H), 6.93 (dd, <com:Math><semantics>J=11.1<annotation encoding="application / x-tex">J = 11.1< / annotation>< / semantics>< / com:Math>, 4.3 Hz, 1H), 6.42 (d, <com:Math><semantics>J=8.0<annotation encoding="application / x-tex">J = 8.0< / annotation>< / semantics>< / com:Math> Hz, 1H), 6.05 (q, <com:Math><semantics>J=6.6<annotation encoding="application / x-tex">J = 6.6< / annotation>< / semantics>< / com:Math> Hz, 1H), 3.92 (s, 3H), 2.29 (s, 3H), 1.80 (d, <com:Math><semantics>J=6.6<annotation encoding="application / x-tex">J = 6.6< / annotation>< / semantics>< / com:Math> Hz, 3H). LC-MS (ESI-MS): 560 [M+H]<com:Sup>+< / com:Sup>.

[0340] Compound II-D-9: the acid added was methanesulfonic acid (29µl, 0.44mmol). The final product II-D-9 was an off-white solid, 262mg with 99.5% yield. <com:Sup>1< / com:Sup>H NMR (500 MHz, DMSO) <com:Math><semantics>δ<annotation encoding="application / x-tex">\delta< / annotation>< / semantics>< / com:Math> 8.14 (s, 1H), 7.84 (t, <com:Math><semantics>J=8.0 Hz<annotation encoding="application / x-tex">J = 8.0 \text{ Hz}< / annotation>< / semantics>< / com:Math>, 1H), 7.75 (d, <com:Math><semantics>J=8.1 Hz<annotation encoding="application / x-tex">J = 8.1 \text{ Hz}< / annotation>< / semantics>< / com:Math>, 1H), 7.65 (t, <com:Math><semantics>J=7.7 Hz<annotation encoding="application / x-tex">J = 7.7 \text{ Hz}< / annotation>< / semantics>< / com:Math>, 2H), 7.47 (t, <com:Math><semantics>J=7.6<annotation encoding="application / x-tex">J = 7.6< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.31 (t, <com:Math><semantics>J=8.1<annotation encoding="application / x-tex">J = 8.1< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.15 (dt, <com:Math><semantics>J=16.3<annotation encoding="application / x-tex">J = 16.3< / annotation>< / semantics>< / com:Math>, 7.8 Hz, 2H), 6.92 (t, <com:Math><semantics>J=7.5 Hz<annotation encoding="application / x-tex">J = 7.5 \text{ Hz}< / annotation>< / semantics>< / com:Math>, 1H), 6.41 (d, <com:Math><semantics>J=7.8 Hz<annotation encoding="application / x-tex">J = 7.8 \text{ Hz}< / annotation>< / semantics>< / com:Math>, 1H), 5.97 (t, <com:Math><semantics>J=6.6 Hz<annotation encoding="application / x-tex">J = 6.6 \text{ Hz}< / annotation>< / semantics>< / com:Math>, 1H), 3.93 (s, 3H), 2.36 (s, 3H), 1.77 (d, <com:Math><semantics>J=6.6<annotation encoding="application / x-tex">J = 6.6< / annotation>< / semantics>< / com:Math> Hz, 3H). LC-MS (ESI-MS): 560 [M+H]<com:Sup>+< / com:Sup>.

[0341] Compound II-D-10: the acid added was citric acid (85mg, 0.44mmol). The final product II-D-10 was an off-white solid, 206mg with 68.5% yield. LC-MS (ESI-MS): 560 <com:Math><semantics>[M+H]+<annotation encoding="application / x-tex">[M+H]^{+}< / annotation>< / semantics>< / com:Math>.

[0342] Compound II-D-11: the acid added was succinic acid (52mg, 0.44mmol). The final product II-D-11 was an off-white solid, 198mg with 73.1% yield. LC-MS (ESI-MS): 560 <com:Math><semantics>[M+H]+<annotation encoding="application / x-tex">[M+H]^{+}< / annotation>< / semantics>< / com:Math>.

[0343] Compound II-D-12: the acid added was L-malic acid (66mg, 0.44mmol). The final product II-D-12 was an off-white solid, 213mg with 76.7% yield. LC-MS (ESI-MS): 560 <com:Math><semantics>[M+H]+<annotation encoding="application / x-tex">[M+H]^{+}< / annotation>< / semantics>< / com:Math>.

[0344] Compound II-D-13: the acid added was D-malic acid (59mg, 0.44mmol). The final product II-D-13 was an off-white solid, 201mg with 72.4% yield. LC-MS (ESI-MS): 560 <com:Math><semantics>[M+H]+<annotation encoding="application / x-tex">[M+H]^{+}< / annotation>< / semantics>< / com:Math>.

[0345] Compound II-D-14: the acid added was L-tartaric acid (66mg, 0.44mmol). The final product II-D-14 was an off-white solid, 209mg with 73.6% yield. LC-MS (ESI-MS): 560 <com:Math><semantics>[M+H]+<annotation encoding="application / x-tex">[M+H]^{+}< / annotation>< / semantics>< / com:Math>.

[0346] Compound II-D-15: the acid added was benzoic acid (54mg, 0.44mmol). The final product II-D-15 was an off-white solid, 202mg with 74% yield. LC-MS (ESI-MS): 560[M+H]<com:Sup>+< / com:Sup>.

[0347] Compound II-D-16: the acid added was maleic acid (51mg, 0.44mmol). The final product II-D-16 was an off-white solid, 220mg with 81.5% yield. LC-MS (ESI-MS): 560[M+H]<com:Sup>+< / com:Sup>.

[0348] Compound II-D-17: the acid added was oxalic acid (55mg, 0.44mmol). The final product II-D-17 was an off-white solid, 216mg with 83.1% yield. LC-MS (ESI-MS): 560[M+H]<com:Sup>+< / com:Sup>. Embodiment 31: Synthesis of Compounds A and B [Image disponible dans le document PDF, Image available in the PDF document]

[0349] Compound 10 (2.0mmol), compound 7 (3.0mmol), potassium carbonate (4.0mmol), tetrakis(triphenylphosphine)palladium(0) (0.02mmol) were placed separately in a 50ml reaction flask with the solvent, 9ml of dioxane and 3ml of water, heated to 100°C with stirring to react for about 14h, and brought to room temperature when the reaction was completed as detected by the TLC test. The reaction liquid was filtered with diatomite and rinsed with ethyl acetate. The filtrate was extracted with 20ml of water and 50ml of ethyl acetate. The ethyl acetate layer was separated, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to obtain a white solid product.

[0350] Compound A (R<com:Sub>2< / com:Sub>=CH(CH<com:Sub>3< / com:Sub>)<com:Sub>2< / com:Sub>): 0.67g, 63% yield. <com:Sup>1< / com:Sup>H NMR (500 MHz, CDCl<com:Sub>3< / com:Sub>) <com:Math><semantics>δ<annotation encoding="application / x-tex">\delta< / annotation>< / semantics>< / com:Math> <com:Math><semantics>8.27 (dt, J=11.9,5.8 Hz, 1H),8.05 (s, 1H),7.80 (ddd, J=11.3,9.6,4.4 Hz, 2H),7.55−7.49 (ddd, J=11.3,9.6,4.4 Hz, 2H),7.55−7.49 (ddd, J=11.3,9.6,4.4<annotation encoding="application / x-tex">8.27 \text{ (dt, } J = 11.9, 5.8 \text{ Hz, } 1\text{H}), 8.05 \text{ (s, } 1\text{H}), 7.80 \text{ (ddd, } J = 11.3, 9.6, 4.4 \text{ Hz, } 2\text{H}), 7.55 - 7.49 \text{ (ddd, } J = 11.3, 9.6, 4.4 \text{ Hz, } 2\text{H}), 7.55 - 7.49 \text{ (ddd, } J = 11.3, 9.6, 4.4< / annotation>< / semantics>< / com:Math> (m, 1H), 7.47 (td, <com:Math><semantics>J=7.7<annotation encoding="application / x-tex">J = 7.7< / annotation>< / semantics>< / com:Math>, 1.1 Hz, 1H), 7.37 (dd, <com:Math><semantics>J=11.5<annotation encoding="application / x-tex">J = 11.5< / annotation>< / semantics>< / com:Math>, 2.0 Hz, 1H), 7.31 (d, <com:Math><semantics>J=7.6<annotation encoding="application / x-tex">J = 7.6< / annotation>< / semantics>< / com:Math> Hz, 2H), <com:Math><semantics>7.16<annotation encoding="application / x-tex">7.16< / annotation>< / semantics>< / com:Math> (t, <com:Math><semantics>J=7.5<annotation encoding="application / x-tex">J = 7.5< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>7.07<annotation encoding="application / x-tex">7.07< / annotation>< / semantics>< / com:Math> (t, <com:Math><semantics>J=8.4<annotation encoding="application / x-tex">J = 8.4< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>6.89<annotation encoding="application / x-tex">6.89< / annotation>< / semantics>< / com:Math> (td, <com:Math><semantics>J=7.8<annotation encoding="application / x-tex">J = 7.8< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>1.1<annotation encoding="application / x-tex">1.1< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>6.50<annotation encoding="application / x-tex">6.50< / annotation>< / semantics>< / com:Math> (d, <com:Math><semantics>J=8.3 Hz<annotation encoding="application / x-tex">J = 8.3 \text{ Hz}< / annotation>< / semantics>< / com:Math>, 1H), 6.00 (q, <com:Math><semantics>J=6.7 Hz<annotation encoding="application / x-tex">J = 6.7 \text{ Hz}< / annotation>< / semantics>< / com:Math>, 1H), 5.55 (brs, 2H), 4.60 (dq, <com:Math><semantics>J=12.1<annotation encoding="application / x-tex">J = 12.1< / annotation>< / semantics>< / com:Math>, 6.1 Hz, 1H), 1.91 <com:Math><semantics>(t,J=8.9 Hz,3H),1.38(dt,J=7.0,3.5 Hz,6H).LC−MS(ESI−MS):536[M+H]+.<annotation encoding="application / x-tex">(t, J = 8.9 \text{ Hz}, 3H), 1.38 (dt, J = 7.0, 3.5 \text{ Hz}, 6H). LC-MS (ESI-MS): 536 [M+H]^+.< / annotation>< / semantics>< / com:Math>

[0351] Compound B (R<com:Sub>2< / com:Sub>=cyclopropyl): 0.49g, 46% yield. <com:Sup>1< / com:Sup>H NMR (500 MHz, CDCl<com:Sub>3< / com:Sub>) δ 8.28 (d, <com:Math><semantics>J=7.9<annotation encoding="application / x-tex">J = 7.9< / annotation>< / semantics>< / com:Math> Hz, 1H), 8.06 (s, 1H), 7.88 – 7.76 (m, 2H), 7.50 (dt, <com:Math><semantics>J=15.1<annotation encoding="application / x-tex">J = 15.1< / annotation>< / semantics>< / com:Math>, 7.2 Hz, 2H), 7.36 (ddd, <com:Math><semantics>J=25.8<annotation encoding="application / x-tex">J = 25.8< / annotation>< / semantics>< / com:Math>, 18.5, 8.0 Hz, 4H), 7.17 (t, <com:Math><semantics>J=7.4<annotation encoding="application / x-tex">J = 7.4< / annotation>< / semantics>< / com:Math> Hz, 1H), 6.89 (t, <com:Math><semantics>J=7.6<annotation encoding="application / x-tex">J = 7.6< / annotation>< / semantics>< / com:Math> Hz, 1H), 6.50 <com:Math><semantics>(d,J=7.8 Hz,1H),6.00(q,J=6.7 Hz,1H),5.51(s,2H),3.91−3.81(m,1H),1.92(d,J=6.7 Hz)<annotation encoding="application / x-tex">(d, J = 7.8 \text{ Hz}, 1\text{H}), 6.00 (q, J = 6.7 \text{ Hz}, 1\text{H}), 5.51 (s, 2\text{H}), 3.91 - 3.81 (m, 1\text{H}), 1.92 (d, J = 6.7 \text{ Hz})< / annotation>< / semantics>< / com:Math> 6.7 Hz, 3H), 0.92 – 0.78 (m, 4H). LC-MS (ESI-MS): 534 [M+H]<com:Sup>+< / com:Sup>. Embodiment 32: Synthesis of Compound C [Image disponible dans le document PDF, Image available in the PDF document]

[0352] Intermediate 9c (1.0mmol) was placed in a 20ml reaction flask with 5ml of DMF, stirred to dissolve at room temperature, then mixed in turn with intermediate 4b (1.1mmol) and potassium carbonate (2.0mmol), heated to 40°C to react for about 15h. When the reaction was completed as detected by the TLC test, ethyl acetate was added to extract, and washed with water. The ethyl acetate layer was separated, dried over anhydrous sodium sulphate and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 0.42g of solid product C with a yield of 78%. <com:Sup>1< / com:Sup>H NMR (500 MHz, DMSO) <com:Math><semantics>δ<annotation encoding="application / x-tex">\delta< / annotation>< / semantics>< / com:Math> 7.92 (brs, 1H), 7.89 – 7.80 (m, 1H), 7.77 (d, <com:Math><semantics>J=7.6<annotation encoding="application / x-tex">J = 7.6< / annotation>< / semantics>< / com:Math> Hz, 1H), 7.62 (dd, <com:Math><semantics>J=16.6<annotation encoding="application / x-tex">J = 16.6< / annotation>< / semantics>< / com:Math>, 7.6 Hz, 2H), <com:Math><semantics>7.48−7.40<annotation encoding="application / x-tex">7.48 - 7.40< / annotation>< / semantics>< / com:Math> (m, 2H), <com:Math><semantics>7.37<annotation encoding="application / x-tex">7.37< / annotation>< / semantics>< / com:Math> (d, <com:Math><semantics>J=8.1<annotation encoding="application / x-tex">J = 8.1< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>7.30<annotation encoding="application / x-tex">7.30< / annotation>< / semantics>< / com:Math> (d, <com:Math><semantics>J=8.5<annotation encoding="application / x-tex">J = 8.5< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>7.11<annotation encoding="application / x-tex">7.11< / annotation>< / semantics>< / com:Math> (d, <com:Math><semantics>J=6.9 Hz<annotation encoding="application / x-tex">J = 6.9 \text{ Hz}< / annotation>< / semantics>< / com:Math>, 1H), <com:Math><semantics>6.86 (d<annotation encoding="application / x-tex">6.86 \text{ (d}< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>J=7.2 Hz<annotation encoding="application / x-tex">J = 7.2 \text{ Hz}< / annotation>< / semantics>< / com:Math>, 1H), <com:Math><semantics>6.34 (d<annotation encoding="application / x-tex">6.34 \text{ (d}< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>J=6.8 Hz<annotation encoding="application / x-tex">J = 6.8 \text{ Hz}< / annotation>< / semantics>< / com:Math>, 1H), <com:Math><semantics>5.91 (d<annotation encoding="application / x-tex">5.91 \text{ (d}< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>J=6.3 Hz<annotation encoding="application / x-tex">J = 6.3 \text{ Hz}< / annotation>< / semantics>< / com:Math>, 1H), 3.89 (s, 3H), 1.74 (d, <com:Math><semantics>J−6.1<annotation encoding="application / x-tex">J - 6.1< / annotation>< / semantics>< / com:Math> Hz, 3H). LC-MS (ESI-MS): 544 [M+H]<com:Sup>+< / com:Sup>. Embodiment 33: Synthesis of Compound D [Image disponible dans le document PDF, Image available in the PDF document]

[0353] Intermediate 9d (1.0mmol) was placed in a 20ml reaction flask with 5ml of DMF, stirred to dissolve at room temperature, then mixed in turn with Intermediate 4c (1.1mmol) and potassium carbonate (2.0mmol), heated to 40°C to react for about 15h. When the reaction was completed as detected by the TLC test, ethyl acetate was added, and washed with water. The ethyl acetate layer was separated, dried over anhydrous sodium sulphate and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 0.32g of solid product D with a yield of 58%. <com:Sup>1< / com:Sup>H NMR (500 MHz, CDCl<com:Sub>3< / com:Sub>) <com:Math><semantics>δ<annotation encoding="application / x-tex">\delta< / annotation>< / semantics>< / com:Math> 8.04 (s, 1H), 7.71 (td, J = 8.2, 5.4 Hz, 1H), 7.61 (d, J = 8.2 Hz, 1H), 7.46 (t, J = 7.7) Hz, 1H), <com:Math><semantics>7.37<annotation encoding="application / x-tex">7.37< / annotation>< / semantics>< / com:Math> (dd, <com:Math><semantics>J=11.4<annotation encoding="application / x-tex">J = 11.4< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>1.9<annotation encoding="application / x-tex">1.9< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>7.30<annotation encoding="application / x-tex">7.30< / annotation>< / semantics>< / com:Math> (dd, <com:Math><semantics>J=13.9<annotation encoding="application / x-tex">J = 13.9< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>8.5<annotation encoding="application / x-tex">8.5< / annotation>< / semantics>< / com:Math> Hz, 2H), <com:Math><semantics>7.15<annotation encoding="application / x-tex">7.15< / annotation>< / semantics>< / com:Math> (dd, <com:Math><semantics>J=15.6<annotation encoding="application / x-tex">J = 15.6< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>8.0 Hz<annotation encoding="application / x-tex">8.0 \text{ Hz}< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>2H<annotation encoding="application / x-tex">2\text{H}< / annotation>< / semantics>< / com:Math>), <com:Math><semantics>7.07 (dd, J=17.1,8.8 Hz<annotation encoding="application / x-tex">7.07 \text{ (dd, J} = 17.1, 8.8 \text{ Hz}< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>1H<annotation encoding="application / x-tex">1\text{H}< / annotation>< / semantics>< / com:Math>), <com:Math><semantics>6.88 (t, J=7.7 Hz<annotation encoding="application / x-tex">6.88 \text{ (t, J} = 7.7 \text{ Hz}< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>1H<annotation encoding="application / x-tex">1\text{H}< / annotation>< / semantics>< / com:Math>), <com:Math><semantics>6.49 (d, J=7.9 Hz<annotation encoding="application / x-tex">6.49 \text{ (d, J} = 7.9 \text{ Hz}< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>1H<annotation encoding="application / x-tex">1\text{H}< / annotation>< / semantics>< / com:Math>), <com:Math><semantics>5.96<annotation encoding="application / x-tex">5.96< / annotation>< / semantics>< / com:Math> (q, J = <com:Math><semantics>6.7<annotation encoding="application / x-tex">6.7< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>5.56<annotation encoding="application / x-tex">5.56< / annotation>< / semantics>< / com:Math> (brs, 2H), <com:Math><semantics>4.60<annotation encoding="application / x-tex">4.60< / annotation>< / semantics>< / com:Math> (dq, J = <com:Math><semantics>12.0<annotation encoding="application / x-tex">12.0< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>6.0<annotation encoding="application / x-tex">6.0< / annotation>< / semantics>< / com:Math> Hz, 1H), <com:Math><semantics>1.89<annotation encoding="application / x-tex">1.89< / annotation>< / semantics>< / com:Math> (d, J = <com:Math><semantics>6.8<annotation encoding="application / x-tex">6.8< / annotation>< / semantics>< / com:Math> Hz, 3H), 1.39 (dd, <com:Math><semantics>J=6.0<annotation encoding="application / x-tex">J = 6.0< / annotation>< / semantics>< / com:Math>, 2.5 Hz, 6H). LC-MS (ESI-MS): 554 [M+H]<com:Sup>+< / com:Sup>. Embodiment 34: CK1 kinase activity assay

[0354] The in-vitro assay of inhibitory activity of the compound herein against CK1ε kinase was:

[0355] (1) preparation of <com:Math><semantics>1×<annotation encoding="application / x-tex">1 \times< / annotation>< / semantics>< / com:Math> kinase basic buffer and stop buffer:

[0356] — 1× kinase basic buffer: 50 mM HEPES, pH 7.5; and 0.0015% Brij-35;

[0357] — stop buffer: 100 mM HEPES, pH 7.5; 0.015% Brij-35; 0.2% Coating Reagent #3; and 50 mM EDTA.

[0358] (2) preparation of the test compound:

[0359] — a 50× ("n×" represents dilution factor, same hereafter) stock solution of the compound with DMSO (10mM, DMSO) to obtain stock solution I for use; 100µl of stock solution I was placed in a 96-well plate, each compound sample was diluted by 5-fold serial dilution till 10 concentrations was obtained while ensuring 10µl of drug solution in each well; meanwhile, 100µl of DMSO was used as a blank control group and a negative control group without enzyme substrate;

[0360] — a separate 96-well plate was prepared, and to 10µl of the above compound,

[0361] 90µl of 1× kinase foundation buffer was added, mixed well for 10min to obtain a mixture.

[0362] (3) preparation of the test plate:

[0363] — 5µl of the above mixture prepared in the 96-well plate was placed in a 384-well plate, each compound in duplicate;

[0364] — a <com:Math><semantics>2.5×<annotation encoding="application / x-tex">2.5 \times< / annotation>< / semantics>< / com:Math> kinase solution was prepared with corresponding <com:Math><semantics>1×<annotation encoding="application / x-tex">1 \times< / annotation>< / semantics>< / com:Math> kinase basic buffer added;

[0365] — a 2.5× polypeptide solution was prepared, and <com:Math><semantics>1×<annotation encoding="application / x-tex">1\times< / annotation>< / semantics>< / com:Math> kinase basic buffer was added with FAM-labelled polypeptide and ATP;

[0366] — <com:Math><semantics>10μl<annotation encoding="application / x-tex">10\mu l< / annotation>< / semantics>< / com:Math> of <com:Math><semantics>2.5×kinase<annotation encoding="application / x-tex">2.5 \times kinase< / annotation>< / semantics>< / com:Math> solution was placed in the test 384-well plate and allowed to stand for 10min at room temperature, mixed with 10<com:Math><semantics>μ<annotation encoding="application / x-tex">\mu< / annotation>< / semantics>< / com:Math>l of 2.5<com:Math><semantics>×<annotation encoding="application / x-tex">\times< / annotation>< / semantics>< / com:Math> polypeptide solution, and reacted for 1h at 28°C and then mixed with 25µl of stop buffer.

[0367] — the plate was read with Caliper program and IC<com:Sub>50< / com:Sub> values were obtained for corresponding compounds in kinase inhibition based on the obtained data. Table 1 Inhibitory activities of partial compounds against CK1ε kinase [Image disponible dans le document PDF, Image available in the PDF document] [Image disponible dans le document PDF, Image available in the PDF document] [Image disponible dans le document PDF, Image available in the PDF document] *: Data from literature values in: WO2017079558A1.

[0368] Data in Table 1 demonstrates that all compounds obtained in the present invention have shown remarkable inhibitory activities against CK1 kinase, which are all significantly better than that of compound CUX-031723 (International Publication No.: WO2017079558A1, IC<com:Sub>50< / com:Sub> of CK1<com:Math><semantics>ϵ<annotation encoding="application / x-tex">\epsilon< / annotation>< / semantics>< / com:Math> is 9.362<com:Math><semantics>μ<annotation encoding="application / x-tex">\mu< / annotation>< / semantics>< / com:Math>M) and compounds A~D (4-amino-disubstituted phenyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl analogues). Wherein, the IC<com:Sub>50< / com:Sub> values for CK1ε inhibition by chiral compounds II-A-1a, II-A-2a, II-A-7a, II-A-8a, and II-B-2a are as high as 108, 88, 45, 60, and 202nM, respectively. The inhibitory activities against CK1 s of compounds II-A-2, II-A-4, and II-A-7 showed a 12-, 12-, and 7.2-fold increase, respectively, compared to compounds A, B, and C, respectively, and the inhibitory activity against CK1 of II-A-16 shows a 12.9- and 93.6-fold increase, respectively, compared to compounds D and CUX-031773, respectively, indicating that the 2-position fluorine atom on the substituent benzene ring in the pyrazolo[3,4-d]pyrimidin-1-yl portion of the compounds of the present invention is very important in improving the CK1ε inhibition activity. It is demonstrated that the compounds of the present invention have further application prospects. Embodiment 35: Determination of in-vitro anti-tumor activity

[0369] The in-vitro anti-tumor activities of the synthesized compounds were determined using a diffuse large B-cell lymphoma (OCI-LY10) cell strain:

[0370] Drug preparation method: The drug was dissolved in DMSO to make a stock solution of 10mM and diluted in a certain ratio to obtain 7 different concentrations (test concentration: <com:Math><semantics>100×<annotation encoding="application / x-tex">100 \times< / annotation>< / semantics>< / com:Math>).

[0371] In vitro culture of tumor cells: The selected OCI-LY10 cells were incubated with a medium of IMDM + fetal bovine serum in a 5% CO<com:Sub>2< / com:Sub> cell culture incubator at 37°C, and were passaged when the cell density grew to 70-90% (adherent cells were passaged after digestion) with Duck's EDTA) for later experiments as needed. Four thousand cells / 200µl / well were seeded in a 96-well plate and incubated overnight in a 5% CO<com:Sub>2< / com:Sub> cell incubator at 37°C. Two μl of the compounds were dispensed in each well at final concentrations of <com:Math><semantics>10μM<annotation encoding="application / x-tex">10\mu M< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>2.5μM<annotation encoding="application / x-tex">2.5\mu M< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>0.625μM<annotation encoding="application / x-tex">0.625\mu M< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>0.15625μM<annotation encoding="application / x-tex">0.15625\mu M< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>0.039063μM<annotation encoding="application / x-tex">0.039063\mu M< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>0.0097656μM<annotation encoding="application / x-tex">0.0097656\mu M< / annotation>< / semantics>< / com:Math> and <com:Math><semantics>0.0024414μM<annotation encoding="application / x-tex">0.0024414\mu M< / annotation>< / semantics>< / com:Math>, and incubation was performed together for 72h in a 5% CO<com:Sub>2< / com:Sub> cell incubator at 37°C with DMSO (2%) as control. After 72h, 20µl of CCK-8 solution was added and the plate was placed in a 5% CO<com:Sub>2< / com:Sub> cell culture incubator at 37°C for 4h. Wells with the corresponding amounts of cell culture medium and CCK-8 solution but without cells were used as blank controls. The absorbance (OD values) was measured at 450nm using an enzyme marker, and the obtained data were used to calculate the IC<com:Sub>50< / com:Sub>. See Table 2 for the test results.

[0372] The cell inhibition rate was calculated by the formula: % cell inhibition = [ (OD of control group - OD of blank group) - (OD of dosing group - OD of blank group) ] / (OD of control group- OD of blank group) × 100%, and the half-maximal inhibitory concentration (IC<com:Sub>50< / com:Sub>) was calculated using the CalcuSyn software. Table 2 Inhibitory activities of partial compounds against OCI-LY10 kinase [Image disponible dans le document PDF, Image available in the PDF document] [Image disponible dans le document PDF, Image available in the PDF document]

[0373] Results show that at the cellular level, the tested compounds exhibited significant tumor cell proliferation inhibitory activities (IC<com:Sub>50< / com:Sub><0.31<com:Math><semantics>μ<annotation encoding="application / x-tex">\mu< / annotation>< / semantics>< / com:Math>M) against the OCI-LY10 cell stain, all significantly better than that of compound D, wherein, the <com:Math><semantics>IC50<annotation encoding="application / x-tex">IC_{50}< / annotation>< / semantics>< / com:Math> value of compound II-A- 7 reached 22nM, showing a 123-fold increase compared to compound C. This further indicates that the 2-position fluorine atom on the substituent benzene ring in the 4- aminopyrazolo[3,4-d]pyrimidin-1-yl portion of the compounds of the present invention is very important in improving the CK1ε inhibition activity. And it is further demonstrated that the compounds of the present invention have broad prospects in anti-tumor applications. Embodiment 36: Determination of in-vitro anti-tumor cells activity

[0374] In-vitro anti-tumor activities of the synthesized compounds were determined using Human Non-Hodgkin's Ki-positive Large Cell Lymphoma (Karpas299) cell strain:

[0375] Drug preparation method: The drugs were dissolved in DMSO to make stock solutions of 10mM and diluted in a certain ratio to obtain 7 different concentrations (test concentration: <com:Math><semantics>100×<annotation encoding="application / x-tex">100\times< / annotation>< / semantics>< / com:Math>).

[0376] In vitro culture of tumor cells: The selected Karpas299 cells were incubated in a 5% CO<com:Sub>2< / com:Sub> cell culture incubator at 37°C using RPMI-1640 + fetal bovine serum medium. Four thousand cells / 100µl / well were seeded in a 96-well plate and incubated overnight in a 5% CO<com:Sub>2< / com:Sub> cell incubator at 37°C. Five µl of the compounds were dispensed in each well at final concentrations of <com:Math><semantics>10μM<annotation encoding="application / x-tex">10\mu M< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>2.5μM<annotation encoding="application / x-tex">2.5\mu M< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>0.625μM<annotation encoding="application / x-tex">0.625\mu M< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>0.15625μM<annotation encoding="application / x-tex">0.15625\mu M< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>0.039063μM<annotation encoding="application / x-tex">0.039063\mu M< / annotation>< / semantics>< / com:Math>, <com:Math><semantics>0.0097656μM<annotation encoding="application / x-tex">0.0097656\mu M< / annotation>< / semantics>< / com:Math> and 0.0024414µM, and incubation was performed together for 72h in a 5% CO<com:Sub>2< / com:Sub> cell incubator at 37°C with DMSO (0.1%) as controls. After 72h, the CellTiter-Glo buffer and reaction substrate were taken out of the fridge and equilibrated to room temperature. The buffer was then poured into a brown bottle containing the substrate. The bottle was turned up and down so that the substrate powder was fully dissolved. The cells were observed under the microscope and the cell culture plate was allowed to equilibrate at room temperature for 30min. The prepared CellTiter-Glo was dispensed in a 96-well plate at 100µl per well. The plate was shaken on a plate shaker for 10min to mix well, and then allowed to stand at room temperature for 10min. A white sealing film was attached to the bottom of the well plate, and the chemiluminescence signal of each well was detected with an Enspire microplate reader. The obtained data were used to calculate the <com:Math><semantics>IC50<annotation encoding="application / x-tex">IC_{50}< / annotation>< / semantics>< / com:Math> values. The test results are shown in Table 3.

[0377] The cell inhibition rate was calculated by the formula: % cell inhibition = [ (OD of control group - OD of blank group) - (OD of dosing group - OD of blank group) ] / (OD of control group- OD of blank group) × 100%, and the half-maximal inhibitory concentration (IC<com:Sub>50< / com:Sub>) was calculated using CalcuSyn software. Table 3 Inhibitory activities of partial compounds against Karpas 299 [Image disponible dans le document PDF, Image available in the PDF document]

[0378] Results show that at the cellular level, the tested compounds exhibited significant tumor cell proliferation inhibitory activities (IC<com:Sub>50< / com:Sub><1 µM) against the Karpas 299 cell stain, further demonstrating that the compounds of the present invention have broad prospects in anti-tumor applications. Embodiment 37: In vivo pharmacodynamic study of ectopic inoculation of OCI-LY10 in SCID mouse metastatic model

[0379] (1) Methods

[0380] OCI-LY10 cells were cultured in an IMDM medium containing 20% FBS and maintained in a 5% CO<com:Sub>2< / com:Sub> incubator at 37°C with saturated humidity. OCI-LY10 cells of logarithmic phase were collected and resuspended in an IMDM basic medium containing 50% Matrigel<com:Sup>TM< / com:Sup>, and the cell concentration was adjusted to 5×10<com:Sup>7< / com:Sup> / ml. Subcutaneous inoculation of 0.1ml of the cell suspension was performed aseptically in the right dorsum of mice at a concentration of <com:Math><semantics>5×106 / 0.1 ml / mouse<annotation encoding="application / x-tex">5 \times 10^6 / 0.1 \text{ ml / mouse}< / annotation>< / semantics>< / com:Math>. Once the tumor volume reached about 150 mm<com:Sup>3< / com:Sup>, the animals were randomly grouped according to tumor volume so that the difference in tumor volume between the groups was less than 10% of the mean value. The date of grouping was recorded as Day 0, and drug administration was started based on the body weight of the animals. During drug administration, body weight of certain animals decreased by more than 15% compared to Day 0 (BWL≥15%), drug administration was discontinued for these animals until their body weight recovered (BWL<15%). The body weight and tumor volume of the animals were measured twice a week during the experiment, and clinical signs were observed and recorded daily.

[0381] (2) Evaluation indicators

[0382] The efficacy in terms of tumor volume was evaluated by the relative tumor proliferation rate T / C ratio, and T / C (%) was calculated as <com:Math><semantics>TRTV / CRTV×100%<annotation encoding="application / x-tex">T_{RTV} / C_{RTV} \times 100\%< / annotation>< / semantics>< / com:Math>, where <com:Math><semantics>TRTV<annotation encoding="application / x-tex">T_{RTV}< / annotation>< / semantics>< / com:Math> is the relative tumor volume (RTV) of the treatment group and <com:Math><semantics>CRTV<annotation encoding="application / x-tex">C_{RTV}< / annotation>< / semantics>< / com:Math> is the RTV of the negative control group. The RTV was calculated as <com:Math><semantics>Vt / V0<annotation encoding="application / x-tex">V_t / V_0< / annotation>< / semantics>< / com:Math>, where <com:Math><semantics>V0<annotation encoding="application / x-tex">V_0< / annotation>< / semantics>< / com:Math> is the tumor volume at the time of grouping and V<com:Sub>t< / com:Sub> is the tumor volume at each measurement. The drug is considered effective with T / C≤40% according to NMPA guidelines.

[0383] The efficacy in terms of tumor weight was evaluated by TGI (tumor weight inhibition) values, TGI %=<com:Math><semantics>(TWC−TWT) / TWC×100%<annotation encoding="application / x-tex">(TW_C-TW_T) / TW_C \times 100\%< / annotation>< / semantics>< / com:Math>, where TW<com:Sub>C< / com:Sub> is tumor weight of control group, and TW<com:Sub>T< / com:Sub> is tumor weight of treatment group. The drug is considered effective with TGI≥58% according to NIH guidelines.

[0384] (3) Results

[0385] The mice performed well in terms of body weight during drug administration, and no drug-related animal death or other significant drug-related toxic or side effects were observed during the experiment. At the end of the last administration, the tumor volume was measured, and the tumors were taken and weighed. See Tables 4 and 5 for the results. Table 4 Effect of test materials on tumor volume and tumor weight in the human diffuse large B-cell lymphoma OCI-LY10 xenograft tumor model (N=5) [Image disponible dans le document PDF, Image available in the PDF document] [Image disponible dans le document PDF, Image available in the PDF document] Table 5 Effect of test materials on tumor volume and tumor weight in the human diffuse large B-cell lymphoma OCI-LY10 xenograft tumor model (N=5) [Image disponible dans le document PDF, Image available in the PDF document] [Image disponible dans le document PDF, Image available in the PDF document]

[0386] According to the results in Tables 4 and 5, the tested compounds showed excellent tumor suppressive effects with T / C less than 40% and TGI greater than 58% in both single and combination groups. In particular, the tumor suppressive effect of the treatment group was remarkable where BTK inhibitor 17a (Patent Application No. 201710998664.5) was administered in combination: 1) II-A-1+17a: T / C of 17.49% and TGI of 83.96%; 2) II-A- 7+17a: T / C of 16.59% and TGI of 86.34%; 3) II-A-1a +17a: T / C of 10.40% and TGI of 88.06%; 4) II-A-7a+17a: T / C of 7.52% and TGI of 90.82%. Therefore, the compounds of the present invention have broad prospects in anti-tumor applications. Embodiment 38: Bioavailability study in rats

[0387] Methods:

[0388] SD rats were used as the experimental animals, to which drugs of 10mg / kg were administered by gavage and 1mg / kg by tail intravenous injection. Blood samples were taken at the following time points for administration by gavage: 0.25, 0.5, 1, 2, 4, 6, 8, and 24h before and after drug administration; time points for blood sampling for intravenous administration were: 0.0833, 0.25, 0.5, 1, 2, 4, 6, 8, and 24h before and after drug administration. Whole blood (0.3ml) was collected, 0.1ml of plasma was separated by centrifugation at 2000g for 10min, and the samples were analyzed by LC-MS / MS.

[0389] Absolute bioavailability was calculated as: <com:Math><semantics>F=(AUCPO / DosePO) / (AUCIV / DoseIV)×I<annotation encoding="application / x-tex">F = (AUC_{PO} / Dose_{PO}) / (AUC_{IV} / Dose_{IV}) \times I< / annotation>< / semantics>< / com:Math> <com:Math><semantics>100%<annotation encoding="application / x-tex">100\%< / annotation>< / semantics>< / com:Math> Table 6 Summary of primary pharmacokinetic parameters after oral administration to rats [Image disponible dans le document PDF, Image available in the PDF document] [Image disponible dans le document PDF, Image available in the PDF document]

[0390] The pharmacokinetic properties of the compound in the free state (II-A-7a) and the pharmaceutically acceptable salt thereof (II-D-6) were investigated in rats, and the results in Table 6 show that the compound of the present invention can be absorbed orally; and the pharmaceutically acceptable salt can improve the oral absorption of the drug by increasing the in vivo exposure of the compound in the free state, with a significant increase in bioavailability (3.7-fold). Therefore, the compounds of the present invention can be administered by oral absorption for treatment of diseases.

Claims

1. A compound having a structure of general formula I: [Image disponible dans le document PDF, Image available in the PDF document] or a stereoisomer thereof, or a stereoisomer mixture thereof, or a pharmaceutically acceptable salt thereof; wherein: R 1 R_1 is H, substituted or unsubstituted C 1 C_1 - C 6 C_6 alkyl, substituted or unsubstituted C 1 C_1 - C 6 C_6 haloalkyl, substituted or unsubstituted C 1 -C 6 alkoxy, substituted or unsubstituted C 1 -C 6 haloalkoxy, substituted or unsubstituted C 2 C_2 - C 10 C_{10} alkenyl, substituted or unsubstituted C 2 C_2 - C 10 C_{10} alkinyl, halogen, cyano, amino, nitro or hydroxy; m is an integer between 1 and 4; and when m is greater than 1, a plurality of R 1 is the same or different, independently of each other; R 2 R_2 is H, substituted or unsubstituted C 1 C_1 - C 6 C_6 alkyl, substituted or unsubstituted C 3 C_3 - C 10 C_{10} cycloalkyl, or substituted or unsubstituted C 5 -C 12 aryl; R 3 is H, substituted or unsubstituted C 1 -C 6 alkyl, substituted or unsubstituted C 1 -C 6 haloalkyl, substituted or unsubstituted C 1 C_1 - C 6 C_6 alkoxy, substituted or unsubstituted C 1 C_1 - C 6 C_6 haloalkoxy, halogen, cyano, amino, nitro or hydroxy; X is CR 4 or N; R 4 is H, substituted or unsubstituted C 1 -C 6 alkyl, substituted or unsubstituted C 1 -C 6 haloalkyl, substituted or unsubstituted C 1 C_1 - C 6 C_6 alkoxy, substituted or unsubstituted C 1 C_1 - C 6 C_6 haloalkoxy, halogen, cyano, amino or hydroxy, and R 3 and R 4 are not H simultaneously; B is selected from substituted or unsubstituted C 1 -C 6 alkyl, substituted or unsubstituted C 3 -C 10 cycloalkyl, substituted or unsubstituted 3- to 10-membered heterocycloalkyl, substituted or unsubstituted C 5 -C 12 aryl, and substituted or unsubstituted 5- to 12-membered heteroaryl; and * denotes a stereocenter.

2. The compound according to claim 1, wherein the compound has a structure of general formula II-A, II-B or II-C: [Image disponible dans le document PDF, Image available in the PDF document] or the stereoisomer thereof, or the stereoisomer mixture thereof, or the pharmaceutically acceptable salt thereof; wherein: R 5 R_5 is H, substituted or unsubstituted C 1 C_1 - C 6 C_6 alkyl, substituted or unsubstituted C 1 C_1 - C 6 C_6 haloalkyl, substituted or unsubstituted C 1 -C 6 alkoxy, substituted or unsubstituted C 1 -C 6 haloalkoxy, substituted or unsubstituted C 1 -C 6 hydroxyalkyl, substituted or unsubstituted C 1 -C 6 halohydroxyalkyl, halogen, cyano, amino, nitro or hydroxy; o is an integer between 1 and 5; R 6 and R 6 ' are independently selected from H, substituted or unsubstituted C 1 -C 6 alkyl, substituted or unsubstituted C 1 C_1 - C 6 C_6 haloalkyl, substituted or unsubstituted C 1 C_1 - C 6 C_6 alkoxy, substituted or unsubstituted C 1 -C 6 haloalkoxy, halogen, cyano, amino and hydroxy; or, R 6 and R 6 ' are joined to form a 3- to 6-membered cycloalkyl or heterocycloalkyl; n is 0, 1 or 2; wherein: when n is 0, Y is CR 7 R 7 ' or NR 8 ; and Z is CHR 7 R 7 ', OH or NHR 8 ; and when n is 1 or 2, Y is CH or N; and Z is CR 7 R 7 ', O or NR 8 ; R 7 and R 7 ' are independently selected from H, substituted or unsubstituted C 1 -C 6 alkyl, substituted or unsubstituted C 1 -C 6 haloalkyl, substituted or unsubstituted C 1 -C 6 alkoxy, substituted or unsubstituted C 1 -C 6 haloalkoxy, substituted or unsubstituted C 3 -C 10 cycloalkyl, substituted or unsubstituted 3- to 10-membered heterocycloalkyl, halogen, cyano, amino and hydroxy; or, R 7 and R 7 ' are joined to form a 3- to 6-membered cycloalkyl or heterocycloalkyl; R 8 is H, substituted or unsubstituted C 1 -C 6 alkyl, substituted or unsubstituted C 3 -C 10 cycloalkyl, substituted or unsubstituted 3- to 10-membered heterocycloalkyl, -C(O)-R 9 or − S ( O ) 2 − R 10 ; -S(O)_2-R_{10}; R 9 is substituted or unsubstituted C 1 -C 6 alkyl, or substituted or unsubstituted C 3 -C 10 cycloalkyl; and R 10 is substituted or unsubstituted C 1 -C 6 alkyl, or substituted or unsubstituted C 3 -C 10 cycloalkyl.

3. The compound according to claim 2, wherein the compound has a structure of general formula III-A, III-B or III-C: [Image disponible dans le document PDF, Image available in the PDF document] or the stereoisomer thereof, or the stereoisomer mixture thereof, or the pharmaceutically acceptable salt thereof.

4. The compound according to claim 3, wherein the compound has a structure of general formula IV-A, IV-B or IV-C: CA Application No. 3,139,285 CPST Ref. 41258 / 00002 CPST Doc: 1378-8018-0760.1 [Image disponible dans le document PDF, Image available in the PDF document] [Image disponible dans le document PDF, Image available in the PDF document] [Image disponible dans le document PDF, Image available in the PDF document] Э or the stereoisomer thereof, or the stereoisomer mixture thereof, or the pharmaceutically acceptable salt thereof; wherein: R 1 is selected from H, substituted or unsubstituted C 1 -C 6 alkyl, and halogen; m is selected from an integer of 1 to 4; R 2 is selected from methyl, ethyl, isopropyl and cyclopropyl; R 5 is selected from H, substituted or unsubstituted C 1 -C 6 alkyl, and halogen; o is selected from an integer of 1 to 5; R 6 and R 6 ' are selected from H, substituted or unsubstituted C 1 -C 6 alkyl, and halogen; Y is selected from N and CH; Z is selected from CH 2 , O and NR 8 ; R 8 is selected from H, substituted or unsubstituted C 1 -C 6 alkyl, and -C(O)-R 9 ; and R 9 R_9 is substituted or unsubstituted C 1 C_1 - C 6 C_6 alkyl.

5. The compound according to claim 4, wherein the compound has a structure of general formula V-A, V-B or V-C: CA Application No. 3,139,285 CPST Ref. 41258 / 00002 CPST Doc: 1378-8018-0760.1 [Image disponible dans le document PDF, Image available in the PDF document] or the stereoisomer thereof, or the stereoisomer mixture thereof, or the pharmaceutically acceptable salt thereof; wherein: the definitions of R 1 , m, R 5 and Z are the same as defined in general formulae IV-A, IV-B and IV-C; and R 2 is methyl or isopropyl.

6. The compound according to any one of claims 1-5, wherein the C denoted by * has a R configuration or an S configuration.

7. The compound according to claim 1, wherein the compound is one of the following: [Image disponible dans le document PDF, Image available in the PDF document] [Image disponible dans le document PDF, Image available in the PDF document] CA Application No. 3,139,285 CPST Ref. 41258 / 00002 CPST Doc: 1378_8018_0760 1 [Image disponible dans le document PDF, Image available in the PDF document] [Image disponible dans le document PDF, Image available in the PDF document] [Image disponible dans le document PDF, Image available in the PDF document] or the stereoisomer thereof, or the stereoisomer mixture thereof, or the pharmaceutically acceptable salt thereof.

8. The compound according to claim 1, wherein the pharmaceutically acceptable salt of the compound is one of the following: [Image disponible dans le document PDF, Image available in the PDF document] [Image disponible dans le document PDF, Image available in the PDF document] or the stereoisomer thereof, or the stereoisomer mixture thereof.

9. The compound according to any one of claims 1-8, wherein the C denoted by * has an S configuration.

10. A compound of the following structure: [Image disponible dans le document PDF, Image available in the PDF document] •11. A compound of the following structure: [Image disponible dans le document PDF, Image available in the PDF document] .

12. A pharmaceutically acceptable solvate of the compound as defined in any one of claims 1-11.

13. A pharmaceutical composition, comprising the compound as defined in any one of claims 1-11, and a pharmaceutically acceptable carrier.

14. Use of the compound as defined in any one of claims 1-11 in preparation of a drug for treating a disease, a disorder or a condition that benefits from an inhibition of activity of a casein kinase 1ε.

15. Use of the compound as defined in any one of claims 1-11 for treating a disease, a disorder or a condition that benefits from an inhibition of activity of a casein kinase 1ε.

16. The use of claim 14 or claim 15, wherein the use is in combination with another drug for treating the disease, the disorder or the condition that benefits from the inhibition of activity of a casein kinase 1ε.

17. The use according to claim 16, wherein the another drug is one or more selected from the following drugs: an antineoplastic drug, an anti-inflammatory agent, an immunosuppressive agent, a steroid and a non-steroidal anti-inflammatory agent.

18. The use according to any one of claim 14-17, wherein the disease is selected from B-cell lymphoma, multiple myeloma, leukemia, lung cancer, breast cancer, prostate cancer, bladder cancer, ovarian cancer, pancreatic cancer, sarcoma, colon cancer, kidney cancer, liver cancer, melanoma, brain tumor, rheumatoid arthritis, systemic lupus erythematosus, multiple sclerosis, inflammatory bowel disease, Crohn's disease, colitis, autoimmune hemolytic anemia, ankylosing spondylitis, pemphigus, urticaria, asthma, optic neuritis, psoriasis, chronic obstructive airway disease, dermatitis and baldness.

19. The use according to claim 18, wherein the B-cell lymphoma is diffuse large B-cell lymphoma, follicular lymphoma, chronic lymphocytic lymphoma, chronic lymphocytic leukemia, B-cell prolymphocytic leukemia, lymphoplasmacytic lymphoma, splenic marginal zone lymphoma, plasma cell myeloma, plasmacytoma, extranodal marginal zone B-cell lymphoma, nodal marginal zone B-cell lymphoma, mantle cell lymphoma, mediastinal large B-cell lymphoma, intravascular large B-cell lymphoma, primary effusion lymphoma, Burkitt's lymphoma / leukemia or pulmonary lymphomatoid granulomatosis.

20. The use according to any one of claims 14-17, wherein the disease is T-cell lymphoma.

21. The use according to claim 20, wherein the T-cell lymphoma is peripheral T-cell lymphoma, anaplastic large cell lymphoma, extranodal NK / T-cell lymphoma, cutaneous peripheral T-cell lymphoma, T-cell prolymphocytic leukemia, angioimmunoblastic T-cell lymphoma, adult T-cell leukemia / lymphoma, hepatosplenic T-cell lymphoma, intestinal T-cell lymphoma, breast implant-associated anaplastic large cell lymphoma, orT-cell large granular lymphocytic leukemia.

22. The use according to claim 17, wherein the antineoplastic drug comprising one or more of the following drugs: a mitotic inhibitor, a tubulin proteolysis inhibitor, an alkylation reagent, an antimetabolite, an insertable antibiotic, an enzyme, a topoisomerase inhibitor, a biological response modifier, an immunomodulator, a BTK inhibitor, a Bcl-2 inhibitor, an anti-CD20 monoclonal antibody, an mTOR inhibitor, an mTORC1 inhibitor, an AKT inhibitor, a PI3K inhibitor, a proteasome inhibitor, an EGFR inhibitor, a VEGFR inhibitor, a CDK inhibitor and a PD-1 / PD-L1 inhibitor.

23. The use according to claim 17, wherein the antineoplastic drug is a BTK inhibitor.

24. An intermediate having a structure as shown in the following formula; [Image disponible dans le document PDF, Image available in the PDF document] wherein: R 2 is H, substituted or unsubstituted C 1 -C 6 alkyl, substituted or unsubstituted C 3 -C 10 cycloalkyl, or substituted or unsubstituted C 5 -C 12 aryl; R 3 is H, substituted or unsubstituted C 1 -C 6 alkyl, substituted or unsubstituted C 1 -C 6 haloalkyl, substituted or unsubstituted C 1 -C 6 alkoxy, substituted or unsubstituted C 1 -C 6 haloalkoxy, halogen, cyano, amino, nitro or hydroxy; X is CR 4 or N; R 4 R_4 is H, substituted or unsubstituted C 1 C_1 - C 6 C_6 alkyl, substituted or unsubstituted C 1 C_1 - C 6 C_6 haloalkyl, substituted or unsubstituted C 1 -C 6 alkoxy, substituted or unsubstituted C 1 -C 6 haloalkoxy, halogen, cyano, amino or hydroxy; and wherein R 3 and R 4 are not H simultaneously.