Methods and compositions for targeting Tregs using CCR8 inhibitors

Abstract

The present invention provides compounds of formula (I) as CCR8 inhibitors, which can be used to treat or prevent cancer using CCR8 inhibitors that target tumor-specific regulatory T cells. Formula (I): JPEG2023540661000478.jpg2877

Description

[Technical field]

[0001] The present invention relates to compounds as CCR8 inhibitors, methods for preparing these compounds, and compositions and uses for treating or preventing cancer using CCR8 inhibitors that target tumor-specific regulatory T cells. [Background technology]

[0002] Chemokines are a family of low molecular weight chemotactic cytokines involved in inflammatory cell recruitment and activation. Chemokines regulate and exert their effects on a wide range of cell functions by binding to chemokine receptors (G protein-coupled receptors), causing chemotaxis and activation of various cell subsets of the immune system. Depending on the location of the N-terminal cysteine ​​residue of the protein, chemokines are classified into various classes, including CC, CXC, CX3C, and XC (Charo et al., 2006, N. Engl. J. Med., 354:610-621). CC chemokines contain a CC motif, where the first two cysteines are not separated by any amino acid, and CXC chemokines contain a CXC motif, where the first two cysteines are separated by random amino acids. The activity of chemokines is mainly mediated by tight binding to leukocyte surface receptors.

[0003] The human body has many built-in mechanisms designed to prevent the development of cancer. In this regard, the immune system is thought to play a key role in eradicating cells with genetic mutations. Thus, cancer cells often evolve to persist without being recognized by cells of the normal immune system. In particular, studies show that elevated levels of regulatory T lymphocytes (sometimes referred to herein as "Tregs") in the peripheral circulation and in the tumor microenvironment underlie immune inhibition in cancer patients. An increased number of Tregs is also identified as a barrier to the successful implementation of cancer immunotherapy.

[0004] CCR8 (CC motif chemokine receptor 8) is expressed primarily in Treg and Th2 cells, but not in Th1 cells (Zingoni et al., 1998, J. Immunol., 161:547-51). + Tregs) expressing CD4 + Foxp3 + A subset of Tregs has been shown to be a major driver of immune inhibition and is important for Treg function and inhibition. Furthermore, CCR8 is a specific marker that is selectively upregulated by tumor-resident Tregs in various tumor types. CCR8 + Numerous reports indicate that an increase in Tregs is favorable for tumor escape mechanisms. Clinically, an increase in Tregs in the tumor microenvironment of various cancer types, such as breast, gastric, ovarian, pancreatic, liver, colon, and pancreatic cancer, is associated with poor prognosis. Mechanistically, Tregs not only have the ability to inhibit various antitumor immune responses but also promote angiogenesis in the tumor microenvironment (Ladoire S et al., 2011, Cancer Immunol Immunother, 60:909-18). CCR8 inhibitors have been shown to prevent tumor growth by decreasing tumor-infiltrating Tregs. Thus, CCR8 is considered as a potential therapeutic target for cancer. In addition, CCR8 is expressed in spinal cord neurons and is also the major source of spinal cord CCL1. CCL1 (also called SCYA1, I-309, TCA3, P500 or SISe) is a well-characterized chemokine of the CC subfamily. It attracts immune cells by interacting with the cell surface chemokine receptor CCR8. The CCL1 / CCR8 nerve signaling pathway also plays an important role in neuropathic pain caused by diabetes and spinal cord injury (M. Zychowska et al., 2017, International Immunopharmacology, 52:261-271). Therefore, the CCL1 / CCR8 axis may be a promising new target for drug development in the treatment of diabetic neuropathy. Summary of the Invention [Problem to be solved by the invention]

[0005] Considering the role of CCR8 in the pathogenic mechanisms of various diseases, there is a need to prepare compounds that inhibit the activity of CCR8, which can be used to treat diseases such as cancer and / or neuropathic pain mediated by CCR8. [Means for solving the problem]

[0006] The present invention provides novel compounds as CCR8 inhibitors, in some embodiments, the present invention provides compounds of formula (I) or a pharma- ceutically acceptable salt, ester, isotope, stereoisomer, tautomer, solvate, prodrug, or combination thereof: Formula (I): JPEG0007653166000001.jpg2877

[0007] where A is a 6-, 9-, 10- or 11-membered carbocyclic or heterocyclic ring; where A is halo, a C1-6 alkyl group, a C1-6 alkyl group substituted with halo, C 1-6 Alkoxy group, C 3-6 Cycloalkyl groups, -COOH, -NH2, -CN, -NHCO(C 1-6 alkyl), -NH(C 1-6 alkyl), -N(C 1-6 alkyl)2, -CONH2, -COO(C 1-6 alkyl), -OCO(C 1-6 alkyl), -CONH(C 1-6 alkyl), CON(C 1-6 and optionally substituted by 0, 1, 2 or 3 substituents independently selected from m is 0 or 1; p is 0 or 1; t is 0 or 1 or 2 or 3; n is 0 or 1, R5 is O or NH; R1 is H or C 1-6 is an alkyl group, R2 and R3 are each independently H, C 1-6 Alkyl groups, -CH2OH, -COOCH3, C 3-6 is selected from a cycloalkyl group, a phenyl group, a heterocyclic ring, or a heteroaryl ring;

[0008] R4 is C 1-6 Alkyl groups, phenyl groups, 4-, 5-, 6-, 7- or 8-membered cycloalkyl groups, 5-membered heteroaryl rings, bridged (C 5-12 ) a cycloalkyl group, JPEG0007653166000002.jpg44160, B1 and B2 are each a 4-, 5-, 6-, 7-, or 8-membered cycloalkyl group, and B3 is each a 5- or 6-membered carbocyclic ring having a saturated or unsaturated hydrocarbon; wherein the carbon atoms of the phenyl group can be substituted by 0, 1 or 2 N, and the 4-, 5-, 6-, 7- or 8-membered cycloalkyl group, 5-membered heteroaryl ring, bridged (C 5-12 ) the carbon atoms of the cycloalkyl group, B1 or B2, can be substituted by 0, 1, or 2 heteroatoms independently selected from N, O, or S, and the carbon atoms of B3 can be substituted by 0, 1, 2, or 3 heteroatoms independently selected from N, O, or S;

[0009] The phenyl group, 4-, 5-, 6-, 7- or 8-membered cycloalkyl group, bridged (C 5-12 ) a cycloalkyl group, JPEG0007653166000003.jpg43160 is a group consisting of halo, C1-6 alkyl group, C1-6 alkyl group substituted with halo, C 3-8 Cycloalkyl groups, -O(C 1-6 alkyl), -OH, -NH2, -N(C 1-6 alkyl)2, -NHCO(C 1-6 alkyl), -NHBoc, -COOH, -COO(C 1-6 alkyl), -COOC(C 1-6 Alkyl)3, -CO(C 1-6 alkyl), -CH2COO(C 1-6alkyl), -CO(CH)qN(C 1-6 alkyl)2, -COCH2NH2, -CH2CON(C 1-6 alkyl)2, -CH2CONH(C 1-6 alkyl), -(C 1-6 alkyl)OH, -COCH2NHBoc, -COCH(CH3)(NHBoc), -(C 1-6 Alkyl)N(C 1-6 Alkyl)2, -(C 1-6 Alkyl)O(C 1-6 alkyl), α-amino acid group, Optionally substituted by 0, 1, 2 or 3 substituents independently selected from: Here, the C 3-8 The carbon atoms of the cycloalkyl group may be substituted by 0, 1 or 2 heteroatoms independently selected from O, N and S;

[0010] or R4 is JPEG0007653166000005.jpg37160, where q is 0 or 1, Y1 is CH or N; Y2 is CH2, O, or NR6, and R6 is H or C 1-6 is an alkyl group,

[0011] Y3 is -CH2OH, -COO(C 1-6 alkyl), -(C 1-6 Alkyl)N(C 1-6 alkyl)2, -N(C 1-6 alkyl)2, X is JPEG0007653166000006.jpg112160-NHCOC(C 1-6 alkyl)3; where r is 0, 1, 2, 3, 4, and R7 is H or C 1-6 is an alkyl group, The above JPEG0007653166000007.jpg38160 is -OH,C 1-6 Alkyl group, C 3-6 Cycloalkyl groups, halo, halo-substituted C1-6 alkyl groups, C 1-6 optionally substituted with 0, 1, 2 or 3 substituents independently selected from alkoxy groups;

[0012] D is a 4-, 5-, 6-, 7- or 8-membered cycloalkyl group other than a phenyl group, or a bridged (C 5-12 ) a cycloalkyl group, wherein the carbon atoms of D can be replaced by 0, 1 or 2 heteroatoms independently selected from N or O, and D is 1-3 Alkyl group, halo, -COOH, C 1-6 optionally substituted with 0, 1 or 2 substituents independently selected from alkoxy groups; E1 is C 3-7 is a cycloalkyl group, and E2 is CF3 or C 3-6 is a cycloalkyl group, wherein the carbon atom of E2 can be substituted by 0 or 1 heteroatom independently selected from N, O or S; The carbon atoms in the ring of JPEG0007653166000008.jpg38160 can be replaced by 0, 1 or 2 heteroatoms independently selected from N, O or S.

[0013] In some embodiments, R4 is C 1-6 Alkyl groups, phenyl groups, 4-, 5-, 6-, 7- or 8-membered cycloalkyl groups, 5-membered heteroaryl rings, bridged (C 5-12 ) a cycloalkyl group, JPEG0007653166000009.jpg44160, B1 and B2 are each a 4-, 5-, 6-, 7-, or 8-membered cycloalkyl group; wherein the carbon atoms of the phenyl group can be substituted by 0, 1 or 2 N, and the 4-, 5-, 6-, 7- or 8-membered cycloalkyl group, 5-membered heteroaryl ring, bridged (C5-12 ) the carbon atoms of the cycloalkyl group, B1 or B2, can be substituted by 0, 1 or 2 heteroatoms independently selected from N, O or S;

[0014] the phenyl group, the 4-, 5-, 6-, 7- or 8-membered cycloalkyl group, 5-12 ) a cycloalkyl group, JPEG0007653166000010.jpg44160 is -F, -Cl, -CF3, -CH2CF3, -NH2, -CH3, -CH2CH3, -(CH2)2CH3, -CH(CH3)2, -C(CH3)3, -COCH2NH2, -CH2CONHCH 3, -CH2CONH(CH3)2, -OCH3, -OCH2CH3, -O(CH2)2CH3, -CH2OCH3, -(CH2)2OCH3, -CH2N(CH3)2, -(CH2)2N(CH3)2, -(CH2)3N(CH3)2, -COCH3, -CO(CH 2)2CH3, -COCH2N(CH3)2, -CO(CH2)2N(CH3)2, -CO(CH2)3N(CH3)2, -COCH2NHBoc, COCH(CH3)(NHBoc), -COOCH3, -COOCH2CH3, -COO(CH2)2CH3, -CO CH2N(CH3)2, -CON(CH3)2, -COOC(CH3)3, -COOH, -(CH2)2OH, -CH2COOCH3, -N(CH3)2, -NHCOCH3, -NHCOCH2CH3, -NHCO(CH2)2CH3, -NHCOOC(CH3)3, Optionally substituted by 0, 1, 2 or 3 substituents independently selected from:

[0015] In some embodiments, JPEG0007653166000012.jpg1831 is It consists of JPEG0007653166000013.jpg37160, JPEG0007653166000014.jpg1831 is It consists of JPEG0007653166000015.jpg21160.

[0016] In some embodiments, the crosslinked (C 5-12 ) Cycloalkyl groups are It consists of JPEG0007653166000016.jpg92160.

[0017] In some embodiments, JPEG0007653166000017.jpg1631 is It consists of JPEG0007653166000018.jpg20160.

[0018] In some embodiments, JPEG0007653166000019.jpg1836 is It consists of JPEG0007653166000020.jpg25160.

[0019] In some embodiments, JPEG0007653166000021.jpg1326 is It consists of JPEG0007653166000022.jpg18160.

[0020] In some embodiments, JPEG0007653166000023.jpg1321 is It consists of JPEG0007653166000024.jpg41160.

[0021] In some embodiments, X is Selected from JPEG0007653166000025.jpg149160JPEG0007653166000026.jpg134160.

[0022] In some embodiments, A is a phenyl group, and the carbon atoms of the phenyl group can be substituted with 0, 1, or 2 N; or A is naphthalene, the carbon atoms of which may be substituted by 0, 1 or 2 N; Or A is JPEG0007653166000027.jpg1825, A1 is a 5- or 6-membered carbocyclic ring, where the carbon atoms of A1 can be replaced by 0, 1 or 2 heteroatoms independently selected from N, O or S;

[0023] Or A is JPEG0007653166000028.jpg1825, A2 is a 5-, 6- or 7-membered heterocyclic ketone, wherein the heteroatom of A2 is 1 or 2 N; When A is naphthalene and n is 1, R2 and R3 are not H at the same time.

[0024] In some embodiments, where A is JPEG0007653166000029.jpg2319, When n is 0, R4 is independently Selected from JPEG0007653166000030.jpg17160,

[0025] B1 is a 4-, 5-, 6-, 7- or 8-membered cycloalkyl group, and B3 is a 5- or 6-membered carbocyclic ring having a saturated or unsaturated hydrocarbon, The carbon atoms of B1 may be substituted with 0, 1, or 2 heteroatoms independently selected from N, O, or S, and the carbon atoms of B3 may be substituted with 0, 1, 2, or 3 heteroatoms independently selected from N, O, or S; JPEG0007653166000031.jpg17160 is halo, C1-6 alkyl group, C 3-6 Cycloalkyl groups, -O(C 1-6 alkyl), -OH, -NH2, -N(C 1-6 and optionally substituted with 0, 1, 2 or 3 substituents independently selected from alkyl).

[0026] In some embodiments, A is a phenyl group, wherein the carbon atoms of the phenyl group can be substituted by 0, 1, or 2 N; or A is naphthalene, wherein the carbon atoms of said naphthalene can be substituted by 0, 1 or 2 N; Or A is JPEG0007653166000032.jpg1825, A1 is a 5- or 6-membered carbocyclic ring, wherein the carbon atoms of A1 can be replaced by 0, 1 or 2 heteroatoms independently selected from N or O; Or A is JPEG0007653166000033.jpg1825, A2 is a 5-, 6- or 7-membered heterocyclic ketone, wherein the heteroatom of A2 is 1 or 2 N;

[0027] When A is naphthalene, R4 is C 1-6 is an alkyl group, or R4 is a 7-membered cycloalkyl group, a bridged (C 5-12 ) a cycloalkyl group, JPEG0007653166000034.jpg19160, B1 and B2 are each a 4-, 5-, 6-, 7- or 8-membered cycloalkyl group;

[0028] 7-membered cycloalkyl group, bridged (C 5-12 ) the carbon atoms of the cycloalkyl group, B1 or B2, can be substituted by 0, 1 or 2 heteroatoms independently selected from N, O or S; The seven-membered cycloalkyl group is 5-12 ) a cycloalkyl group, JPEG0007653166000035.jpg19160 is C 1-6 Alkyl group, -O(C 1-6 alkyl), -(C 1-6 Alkyl)N(C 1-6 alkyl)2, -CH2CON(C 1-6Alkyl)2, -CO(C 1-6 alkyl), -CO(CH2)qN(C 1-6 alkyl)2, Optionally substituted by 0, 1, 2 or 3 substituents independently selected from:

[0029] or R4 is JPEG0007653166000037.jpg39160, where q is 0 or 1,

[0030] Y1 is CH or N; Y2 is CH2, O or NR6, R6 is H or C 1-6 is an alkyl group, X is JPEG0007653166000038.jpg51160 is selected.

[0031] where A is independently Selected from JPEG0007653166000039.jpg60160, Y is CH or N.

[0032] In some embodiments, A is m is 0 or 1, A is If the image is JPEG0007653166000041.jpg39160, then m is 0.

[0033] In some embodiments, A is, Selected independently from JPEG0007653166000042.jpg39160, Where R4 is C 1-6 Alkyl groups, phenyl groups, 4-, 5-, 6-, 7- or 8-membered cycloalkyl groups, 5-membered heteroaryl rings, bridged (C 5-12 ) a cycloalkyl ring, JPEG0007653166000043.jpg39160,

[0034] B1 and B2 are each a 4-, 5-, 6-, 7-, or 8-membered cycloalkyl group, and B3 is each a 5-, 6-membered carbocyclic ring having a saturated or unsaturated hydrocarbon; wherein the carbon atoms of the phenyl group can be substituted by 0, 1 or 2 N, and the 4-, 5-, 6-, 7- or 8-membered cycloalkyl group, 5-membered heteroaryl ring, bridged (C 5-12 ) the carbon atoms of the cycloalkyl group, B1 or B2, can be substituted by 0, 1, or 2 heteroatoms independently selected from N, O, or S, and the carbon atoms of B3 can be substituted by 0, 1, 2, or 3 heteroatoms independently selected from N, O, or S; The phenyl group, 4-, 5-, 6-, 7- or 8-membered cycloalkyl group, bridged (C 5-12 ) a cycloalkyl group, JPEG0007653166000044.jpg44160 is halo, C1-6 alkyl group, C 3-6 Cycloalkyl groups, -O(C 1-6 alkyl), -NH2, -N(C 1-6 alkyl)2, -NHCO(C 1-6 alkyl), -NHBoc, -COOH, -COO(C 1-6 alkyl), -COOC(C 1-6 Alkyl)3, -CO(C 1-6 alkyl), -CH2COO(C 1-6 alkyl), -CO(CH2)qN(C 1-6 alkyl)2, -COCH2NH2, -CH2CON(C 1-6 alkyl)2, -CH2CONH(C 1-6 alkyl), -(C 1-6 alkyl)OH, -COCH2NHBoc, -COCH(CH3)(NHBoc), -(C 1-6 Alkyl)N(C 1-6 alkyl)2, -CH2CF3, -(C 1-6 Alkyl)O(C 1-6 alkyl), Optionally substituted by 0, 1, 2 or 3 substituents independently selected from:

[0035] Here, C 3-6 The carbon atoms of the cycloalkyl group can be substituted by 0 or 1 O; or R4 is JPEG0007653166000046.jpg34160, where q is 0 or 1,

[0036] Y1 is CH or N; Y2 is CH2, O or NR6, R6 is H or C 1-6 is an alkyl group, Y3 is -CH2OH, -COO(C 1-6 alkyl), -(C 1-6 Alkyl)N(C 1-6 alkyl)2, -N(C 1-6 alkyl)2, X is JPEG0007653166000047.jpg109160-NHCOC(C 1-6 alkyl)3;

[0037] where r is 0, 1, 2, 3, 4, and R7 is H or C 1-6 is an alkyl group, JPEG0007653166000048.jpg35160 is -OH,C 1-6 Alkyl groups, halo, halo-substituted C1-6 alkyl groups, C 1-6 optionally substituted with 0, 1, 2 or 3 substituents independently selected from alkoxy groups; D is a 4-, 5-, 6-, 7- or 8-membered cycloalkyl group other than a phenyl group, or a bridged (C 5-12 ) a cycloalkyl group, wherein the carbon atoms of D can be replaced by 0, 1 or 2 heteroatoms independently selected from N or O, and D is 1-3 Alkyl group, halo, -COOH, C1-6 optionally substituted with 0, 1 or 2 substituents independently selected from alkoxy groups; E1 is C 3-7 is a cycloalkyl group, and E2 is CF3 or C 3-6 is a cycloalkyl group,

[0038] wherein the carbon atom of E2 can be substituted by 0 or 1 heteroatom independently selected from N, O or S; The carbon atoms in the ring of JPEG0007653166000049.jpg35160 can be replaced by 0, 1 or 2 heteroatoms independently selected from N, O or S.

[0039] In some embodiments, wherein R4 is -CHC2H6, -CC3H9; or R4 is JPEG0007653166000050.jpg32160.

[0040] In some embodiments, R4 is C 1-6 Alkyl groups, phenyl groups, 4-, 5-, 6-, 7- or 8-membered cycloalkyl groups, bridged (C 5-12 ) a cycloalkyl group, JPEG0007653166000051.jpg20160, B1 and B2 are each a 5-, 6-, or 7-membered cycloalkyl group; wherein the carbon atoms of the phenyl group can be substituted by 0, 1 or 2 N; and wherein the 4-, 5-, 6- or 7-membered cycloalkyl group, bridged (C 5-12 ) the carbon atoms of the cycloalkyl group, B1 or B2, can be substituted by 0, 1 or 2 heteroatoms independently selected from N, O or S;

[0041] the phenyl group, the 4-, 5-, 6-, 7- or 8-membered cycloalkyl group, 5-12 ) a cycloalkyl group, JPEG0007653166000052.jpg20160 is C 1-6 Alkyl groups, C1-6 alkyl groups substituted with halo, C 3-8 Cycloalkyl groups, -COO(C 1-6 alkyl), -COOH, -COOC(C 1-6 alkyl), -COCH2NH2, -CO(C 1-6 alkyl), -CO(CH)qN(C 1-6 Alkyl)2, -(C 1-6 Alkyl)O(C 1-6 alkyl), -CH2CONH(C 1-6 alkyl), -CH2CON(C 1-6 Alkyl)2, -(C 1-6 alkyl)OH, -CH2COO(C 1-6 alkyl), Optionally substituted by 0, 1, 2 or 3 substituents independently selected from:

[0042] Here, the C 3-8 The carbon atoms of the cycloalkyl group may be substituted by 0, 1 or 2 heteroatoms independently selected from O, N and S; or R4 is unsubstituted JPEG0007653166000054.jpg15160, q is 0 or 1; Y3 is -CH2OH, -COO(C 1-6 alkyl), -(C 1-6 Alkyl)N(C 1-6 alkyl)2, -N(C 1-6 alkyl)2, X is JPEG0007653166000055.jpg81160-NHCOC(C 1-6 alkyl)3;

[0043] where r is 0, 1, 2, 3, 4, and R7 is H or C 1-6 is an alkyl group, The above JPEG0007653166000056.jpg35160 is -OH,C 1-6 Alkyl groups, halo, halo-substituted C1-6 alkyl groups, C 1-6 optionally substituted with 0, 1, 2 or 3 substituents independently selected from alkoxy groups;

[0044] D is a 4-, 5-, 6- or 7-membered cycloalkyl group other than a phenyl group, or a bridged (C 5-12 ) a cycloalkyl group, wherein the carbon atoms of D can be replaced by 0 or 1 heteroatoms independently selected from N or O, and D is 1-3 Alkyl group, halo, -COOH, C 1-6 optionally substituted with 0, 1 or 2 substituents independently selected from alkoxy groups; The carbon atoms in the ring of JPEG0007653166000057.jpg17160 can be replaced by 0, 1 or 2 heteroatoms independently selected from N, O or S.

[0045] In some embodiments, where A is JPEG0007653166000058.jpg1111, Wherein A is halo, C alkyl group, C 1-6 may be optionally substituted by 0, 1, 2 or 3 substituents independently selected from alkoxy groups; Y is CH or N; R1 is H or C 1-6 is an alkyl group, R2 and R3 are each independently H, C 1-6 Alkyl groups, -CH2OH, -COOCH3, C 3-6 cycloalkyl group, phenyl group,

[0046] R4 is C 1-6 Alkyl groups, phenyl groups, 4-, 5-, 6-, 7- or 8-membered cycloalkyl groups, 5-membered heteroaryl rings, bridged (C 5-12 ) a cycloalkyl ring, JPEG0007653166000059.jpg39160, B1 and B2 are each a 4-, 5-, 6-, 7-, or 8-membered cycloalkyl group, and B3 is each a 5- or 6-membered carbocyclic ring having a saturated or unsaturated hydrocarbon; wherein the carbon atoms of the phenyl group can be substituted by 0 or 1 N, and the 4-, 5-, 6-, 7- or 8-membered cycloalkyl group, 5-membered heteroaryl ring, bridged (C 5-12 ) the carbon atoms of the cycloalkyl group, B1 or B2, can be substituted by 0, 1, or 2 heteroatoms independently selected from N, O, or S, and the carbon atoms of B3 can be substituted by 0, 1, 2, or 3 heteroatoms independently selected from N, O, or S;

[0047] the phenyl group, the 4-, 5-, 6-, 7- or 8-membered cycloalkyl group, 5-12 ) a cycloalkyl group, JPEG0007653166000060.jpg44160 is halo, C1-6 alkyl group, C 3-6 Cycloalkyl groups, -O(C 1-6 alkyl), -NH2, -N(C 1-6 alkyl)2, -NHCO(C 1-6 alkyl), -NHBoc, -COOH, -COO(C 1-6 alkyl), -COOC(C 1-6 Alkyl)3, -CO(C 1-6 alkyl), -CH2COO(C 1-6 alkyl), -CO(CH2)qN(C 1-6 alkyl)2, -COCH2NH2, -CH2CON(C 1-6 alkyl)2, -CH2CONH(C 1-6 alkyl), -(C 1-6 alkyl)OH, -COCH2NHBoc, -COCH(CH3)(NHBoc), -(C 1-6 Alkyl)N(C 1-6 alkyl)2, -CH2CF3, -(C 1-6 Alkyl)O(C 1-6 alkyl), Optionally substituted by 0, 1, 2 or 3 substituents independently selected from:

[0048] Here, the C 3-6 The carbon atoms of the cycloalkyl group can be substituted by 0 or 1 O; or R4 is JPEG0007653166000062.jpg34160, where q is 0 or 1,

[0049] Y1 is CH or N; Y2 is CH2, O or NR6, R6 is H or C 1-6 is an alkyl group, Y3 is -CH2OH, -COO(C 1-6 alkyl), -(C 1-6 Alkyl)N(C 1-6 alkyl)2, -N(C 1-6 alkyl)2, X is JPEG0007653166000063.jpg109160-NHCOC(C 1-6 alkyl)3;

[0050] where r is 0, 1, 2, 3, 4, and R7 is H or C 1-6 is an alkyl group, The above JPEG0007653166000064.jpg35160 is -OH,C 1-6 Alkyl groups, halo, halo-substituted C1-6 alkyl groups, C 1-6 optionally substituted with 0, 1, 2 or 3 substituents independently selected from alkoxy groups; D is a 4-, 5-, 6-, 7- or 8-membered cycloalkyl group other than a phenyl group, or a bridged (C 5-12 ) a cycloalkyl group, wherein the carbon atoms of D can be replaced by 0, 1 or 2 heteroatoms independently selected from N or O, and D is 1-3Alkyl group, halo, -COOH, C 1-6 optionally substituted with 0, 1 or 2 substituents independently selected from alkoxy groups;

[0051] E1 is C 3-7 is a cycloalkyl group, and E2 is CF3 or C 3-6 is a cycloalkyl group, wherein the carbon atom of E2 can be substituted by 0 or 1 heteroatom independently selected from N, O or S; The carbon atoms in the ring of JPEG0007653166000065.jpg35160 can be replaced by 0, 1 or 2 heteroatoms independently selected from N, O or S.

[0052] In some embodiments, wherein R4 is C 1-6 an alkyl group, a phenyl group, or a 4-, 5-, 6-, or 7-membered cycloalkyl group; wherein the carbon atoms of the phenyl group may be substituted by 0 or 1 N; and the carbon atoms of the 4-, 5-, 6- or 7-membered cycloalkyl group, B1 or B2, may be substituted by 0, 1 or 2 heteroatoms independently selected from N or O; The phenyl group, 4-, 5-, 6- or 7-membered cycloalkyl group is 1-6 Alkyl groups, -COOH, -COOC(C 1-6 Alkyl)3(Boc), -CO(C 1-6 alkyl), -CH2COO(C 1-6 alkyl), -CO(CH)qN(C 1-6 alkyl)2, -CH2CONH(C 1-6 alkyl), -(C 1-6 Alkyl)OH, -CH2CF3, -(C 1-6 Alkyl)O(C 1-6 alkyl), Optionally substituted by 0, 1, 2 or 3 substituents independently selected from:

[0053] q is 0 or 1; or R4 is JPEG0007653166000067.jpg19160, Y3 is -CH2OH, -COO(C 1-6 alkyl), -(C 1-6 Alkyl)N(C 1-6 alkyl)2, -N(C 1-6 alkyl)2,

[0054] X is JPEG0007653166000068.jpg82160-NHCOC(C 1-6 alkyl)3; where R7 is H or C 1-6 is an alkyl group,

[0055] The above JPEG0007653166000069.jpg21160 is -OH,C 1-3 Alkyl groups, halo, halo-substituted C1-6 alkyl groups, C 1-6 optionally substituted with 0, 1 or 2 substituents independently selected from alkoxy groups; D is a 4-, 5-, 6- or 7-membered cycloalkyl group other than a phenyl group, and the carbon atoms of D can be replaced by 0 or 1 heteroatom independently selected from N or O, and D is C 1-3 Alkyl group, halo, -COOH, C 1-6 optionally substituted with 0, 1 or 2 substituents independently selected from alkoxy groups; The carbon atoms in the ring of JPEG0007653166000070.jpg15160 can be replaced by 0 or 1 heteroatom independently selected from N or S.

[0056] In some embodiments, wherein A is, Selected independently from JPEG0007653166000071.jpg56160, n is 1 and p is 0, R1 is H, R2 and R3 are each independently selected from H or -CH3.

[0057] In some embodiments, Here, A is JPEG0007653166000072.jpg1429, m is 0 or 1; X is Selected from TIFF0007653166000073.tif17160, R4 is, The image is JPEG0007653166000074.jpg1421.

[0058] In some embodiments, where A is JPEG0007653166000075.jpg1432, m is 1, X is JPEG0007653166000076.jpg1321, R4 is, The image is JPEG0007653166000077.jpg1421.

[0059] In some embodiments, where A is JPEG0007653166000078.jpg15160, m is 0, R4 is, The image is JPEG0007653166000079.jpg1421.

[0060] In some embodiments, where A is JPEG0007653166000080.jpg19160, R4 is, The image is JPEG0007653166000081.jpg1321.

[0061] In some embodiments, where A is JPEG0007653166000082.jpg16160, R4 is The image is JPEG0007653166000083.jpg1321.

[0062] In some embodiments, wherein the compound is selected from the following: JPEG0007653166000084.jpg204160JPEG0007653166000085.jpg228160JPEG0007653166000086.jpg212160JPEG0007653166000087.jpg198160JPEG0007653166000088.jpg211160JPEG0007653166000089.jpg232160JPEG0007653166000090.jpg206160JPEG0007653166000091.jpg220160JPEG0007653166000092.jpg215160JPEG0007653166000093.jpg214160JPEG0007653166000094.jpg211160JPEG0007653166000095.jpg205160JPEG0007653166000096.jpg226160JPEG0007653166000097.jpg229160JPEG0007653166000098.jpg211160JPEG0007653166000099.jpg227160JPEG0007653166000100.jpg200160JPEG0007653166000101.jpg220160JPEG0007653166000102.jpg227160JPEG0007653166000103.jpg219160JPEG0007653166000104.jpg212160JPEG0007653166000105.jpg221160JPEG0007653166000106.jpg227160JPEG0007653166000107.jpg217160JPEG0007653166000108.jpg217160JPEG0007653166000109.jpg219160JPEG0007653166000110.jpg218160JPEG0007653166000111.jpg228160JPEG0007653166000112.jpg211160JPEG0007653166000113.jpg228160JPEG0007653166000114.jpg232160JPEG0007653166000115.jpg227160JPEG0007653166000116.jpg218160JPEG0007653166000117.jpg212160JPEG0007653166000118.jpg212160JPEG0007653166000119.jpg207160JPEG0007653166000120.jpg212160JPEG0007653166000121.jpg232160JPEG0007653166000122.jpg210160JPEG0007653166000123.jpg228160JPEG0007653166000124.jpg212160JPEG0007653166000125.jpg232160JPEG0007653166000126.jpg210160JPEG0007653166000127.jpg212160JPEG0007653166000128.jpg227160JPEG0007653166000129.jpg232160JPEG0007653166000130.jpg212160JPEG0007653166000131.jpg207160JPEG0007653166000132.jpg227160JPEG0007653166000133.jpg212160JPEG0007653166000134.jpg212160JPEG0007653166000135.jpg212160JPEG0007653166000136.jpg229160JPEG0007653166000137.jpg220160JPEG0007653166000138.jpg212160JPEG0007653166000139.jpg212160JPEG0007653166000140.jpg212160JPEG0007653166000141.jpg212160JPEG0007653166000142.jpg229160JPEG0007653166000143.jpg207160JPEG0007653166000144.jpg212160JPEG0007653166000145.jpg212160JPEG0007653166000146.jpg212160JPEG0007653166000147.jpg232160JPEG0007653166000148.jpg79160.

[0063] In some embodiments, the present invention provides a pharmaceutical composition comprising a compound of formula I, or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable vector or excipient. In some embodiments, the present invention provides for the use of the compounds in the treatment or prevention of cancer using CCR8 inhibitors that target tumor-specific regulatory T cells. In some embodiments, the present invention provides a method for ameliorating disease symptoms in a subject having a disease characterized by aberrant or unwanted activity of regulatory T cells, the method comprising administering to the subject a therapeutically effective amount of a compound or a pharmaceutical composition thereof. In some embodiments, the present invention provides a method for ameliorating disease symptoms in a subject having a disease mediated by an abnormal or unwanted CCR8 / CCL1 axis, comprising administering to the subject a therapeutically effective amount of a compound or a pharmaceutical composition thereof.

[0064] In some embodiments, the disease state of the method is selected from the group consisting of cancer. In some embodiments, the cancer comprises leukemia. In some embodiments, leukemia includes chronic lymphocytic leukemia, chronic myelogenous leukemia, acute myeloid leukemia, acute lymphoblastic leukemia, and lymphoma leukemia stages. In some embodiments, the cancer comprises a solid tumor. In some embodiments, solid tumors include breast cancer, gastric cancer, colorectal cancer, ovarian cancer, pancreatic cancer and liver cancer. In some embodiments, the condition is selected from the group consisting of neuropathic pain. In some embodiments, the neuropathic pain is caused by diabetes or spinal cord injury. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0065] definition In the context of the present invention, the following definitions apply. The term "CCR8 inhibitors" includes all possible isomers, stereoisomers, enantiomers, diastereomers, tautomers, pharma- ceutically acceptable salts, hydrates, solvates and prodrugs of the CCR8 inhibitors described herein. The article "a / an" is used in this disclosure to refer to one or to more than one (i.e., at least one) of the grammatical object of the article. By way of example, "an element" refers to one element or to one or more elements.

[0066] Unless otherwise stated, the term "and / or" is used in this disclosure to refer to "and" or "or." The term "alkyl group" refers to a monoradical straight or branched saturated hydrocarbon chain. In some embodiments, an alkyl group as used herein is an alkyl group having 1 to 20 carbon atoms (i.e., C 1-20 alkyl group), 1 to 6 carbon atoms (i.e., C 1-6 Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, n-butyl, s-butyl, t-butyl, pentyl, 2-pentyl, isopentyl, neopentyl, hexyl, 2-hexyl, 3-hexyl, and 3-methylpentyl. When an alkyl group residue having a particular number of carbon atoms is named, all geometric isomers having that number of carbon atoms are included, thus, for example, "butyl" can include n-butyl, s-butyl, isobutyl, and t-butyl, and "propyl" can include n-propyl and isopropyl. In some embodiments, a "lower alkyl group" refers to an alkyl group having 1 to 6 carbons (i.e., C 1-6 It refers to an alkyl group.

[0067] Alone or in combination with "C 1-6The term "alkyl group" refers to a straight or branched chain alkyl group having from 1 to 6 carbon atoms. Examples of C1-C6 alkyl groups include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, t-butyl, isopentyl, and neopentyl groups. The term "alkoxy" refers to the group RO-, where R is an alkyl group or -YZ, where Y is an alkylene group and Z is an alkenyl or alkynyl group, where the alkyl, alkenyl, and alkynyl groups are as defined herein. In some embodiments, the alkoxy group is the alkyl group -O-, including, for example, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, t-butoxy, s-butoxy, n-pentyloxy, n-hexyloxy, 1,2-dimethylbutoxy, and the like.

[0068] Alone or in combination with "C 1-6 The term "alkoxy group" refers to the group R'-O-, where R' is C 1-6 It is an alkyl group. 1-6 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, s-butoxy, t-butoxy, pentyloxy, isopentyloxy, neopentyloxy, t-pentyloxy, and hexyloxy groups. The term "cycloalkyl group" refers to a cyclized alkyl group, including monocyclic, bicyclic or polycyclic systems. 3-7 The term "cycloalkyl group" refers to C3, C4, C5, C6 and C7 cycloalkyl groups. Exemplary cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and norbornyl groups. The definition of "cycloalkyl group" includes branched chain cycloalkyl groups such as 1-methylcyclopropyl and 2-methylcyclopropyl groups. "C3-6 The term "cycloalkyl group" refers to a cyclic hydrocarbon containing 3 to 6 carbon atoms. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl groups. It is understood that any replaceable hydrogen on a cycloalkyl group can be replaced by a halo, a C1-C3 alkyl group, a hydroxyl group, an alkoxy group, and a cyano group.

[0069] The term "heterocycle" refers to a cyclic hydrocarbon containing 3 to 10 atoms, where at least one atom is O, N, or S, and where monocyclic heterocycles may contain up to two double bonds. Examples of heterocycles include, but are not limited to, aziridine, ethylene oxide, thietane, azetidine, oxetane, thietane, pyrrolidine, tetrahydrofuran, tetrahydrothiophene, piperidine, tetrahydropyran, thia, imidazolidine, oxazolidine, thiazolidine, dioxolane, dithiolane, piperazine, oxazine, dithiane, and dioxane. The term "halogen" or "halo" refers to fluorine, chlorine, bromine or iodine. Preferred "halogen" groups are fluorine, chlorine or bromine. As used herein, the term "ketone" refers to a compound of formula A 1 C(O)A 2 where A 1 and A 2 can be independently an alkyl group, a cycloalkyl group, an alkenyl group, a cycloalkenyl group, an alkynyl group, a cycloalkynyl group, an aryl group, or a heteroaryl group.

[0070] "Optionally" or "optionally" means that the subsequently described event or circumstance may or may not occur, and the description includes instances when the event or circumstance occurs and instances when the event or circumstance does not occur. "Substituted" groups include embodiments in which a monoradical substituent is attached to a single atom of the substituted group (e.g., forming a branched chain) and also include embodiments in which a substituent may be a diradical bridger attached to two adjacent atoms of the substituted group, thereby forming a fused ring on the substituent. When a given group (moiety) is described herein as being attached to a second group, and the site of attachment is ambiguous, the given group may be attached to the second group at any available site on the given group. For example, "C 1-6 "Phenyl group substituted with alkyl groups" refers to a C-substituted alkyl group attached to any available site on the phenyl group. 1-6 It may be any available site of attachment of the alkyl group. In this regard, an "available site" is a site on the group at which a hydrogen of the group can be replaced by a substituent.

[0071] "Pharmaceutically acceptable salts" refers to salts that retain the biological effectiveness and properties of the free bases obtained by reaction with inorganic acids such as hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid, or by reaction with organic acids such as sulfonic acid, carboxylic acid, organophosphoric acid, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, citric acid, fumaric acid, maleic acid, succinic acid, benzoic acid, salicylic acid, lactic acid, tartaric acid (e.g., (+) or (-)-tartaric acid or its bludgeon), amino acids (e.g., (+) or (-)-amino acids or mixtures thereof). These salts can be prepared by methods known to those skilled in the art. "Pharmaceutically acceptable salts" refers to salts that are suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, and the like, within the scope of medical judgment, and are compatible with the action of the drug commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, SM Berge et al. describe pharma- ceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, which is incorporated herein by reference.

[0072] The term "vector" as used in this disclosure includes vectors, excipients and diluents and refers to a material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, that is involved in carrying or transporting a drug from one organ or part of the body to another organ or part of the body. The term "effective amount" refers to an amount of the compound of the present invention that is sufficient to achieve a desired effect or result in the context of its administration or use. Depending on the context, the term effective amount can include a pharma- ceutically effective amount or a therapeutically effective amount or equivalents. Effective amounts can be determined by methods known to those skilled in the art. A compound of a given formula (e.g., a compound of formula I, including all other formulae herein) refers to the compounds of the present disclosure, as well as pharma- ceutically acceptable salts, pharma-ceutically acceptable esters, isomers, tautomers, solvates, isotopes, hydrates, polymorphs, and prodrugs of these compounds. Additionally, compounds of the present disclosure may have one or more asymmetric centers and may be produced as racemic mixtures or as individual enantiomers or diastereomers. The number of stereoisomers present in a particular compound of a particular formula depends on the number of asymmetric centers present (2n stereoisomers may exist, where n is the number of asymmetric centers). Single stereoisomers may be obtained by resolving racemic or non-racemic mixtures of intermediates at some appropriate stage of the synthesis, or by resolving the compounds by conventional methods.

[0073] Unless otherwise specified, both single stereoisomers (including single enantiomers and diastereomers) and racemic and non-racemic mixtures of stereoisomers are included within the scope of the disclosure, all of which are included within the structures herein. "Isomers" are different compounds that have the same molecular formula. Isomers include stereoisomers, enantiomers, and diastereomers. "Stereoisomers" refer to isomers that differ only in the arrangement of the atoms in space. "Enantiomers" are a pair of stereoisomers that are non-superimposable mirror images of each other. A 1:1 mixture of a pair of enantiomers is a "racemic" mixture. Where appropriate, the term "(±)" is used to designate a racemic mixture. "Diastereomers" are stereoisomers that have at least two asymmetric atoms, but which are not mirror-images of each other.

[0074] Some compounds exist in the form of tautomers. Tautomers are in parallel with each other. For example, amide-containing compounds can exist in equilibrium with imidic acid tautomers. Regardless of which tautomer is shown, and regardless of the nature of the equilibrium between tautomers, those skilled in the art should understand that these compounds include amide tautomers and imidic acid tautomers. Thus, amide-containing compounds are understood to include their imidic acid tautomers. Similarly, imidic acid-containing compounds are understood to include their amide tautomers. The term "solvate" refers to a complex formed by combining a compound of the invention with a solvent. The term "hydrate" refers to a complex formed by combining a compound of the present invention with water. "Prodrug" refers to a compound of the invention that contains a chemical group that can be transformed in vivo and / or separated from the remainder of the molecule to provide the active drug, a pharma- ceutically acceptable salt thereof, or a biologically active metabolite thereof.

[0075] A "subject" is a mammal, such as a human, mouse, rat, guinea pig, dog, cat, horse, cow, pig, or a non-human primate, such as a monkey, chimpanzee, baboon, or rhesus monkey, and the terms "subject" and "patient" are used interchangeably herein. The terms "inhibition" / "inhibit" / "inhibiting" refer to a significant decrease in the baseline activity of a biological activity or process. Unless otherwise specified, the term "disease" as used in this disclosure refers to, and is used interchangeably with, the terms disorder, symptom, or disease. The term "tumor," as used herein, refers to an abnormal growth of tissue. Tumors may be benign or malignant. Malignant tumors are usually referred to as cancers. What distinguishes cancers from benign tumors is the ability of malignant cells to invade other tissues, either by growing directly into adjacent tissues through invasion, or by implanting at distant sites through metastasis (i.e., transport through the blood or lymphatic system).

[0076] The term "inflammatory disease" as used herein refers to a disease that is brought about, caused by, or caused by inflammation. The term "inflammatory disease" may also refer to a dysregulated inflammatory response that causes an overreaction of macrophages, granulocytes, and / or T lymphocytes, resulting in abnormal tissue damage and / or cell death. Inflammatory diseases can be acute or chronic inflammation and can be caused by infectious or non-infectious causes. The inflammatory disease is preferably multiple sclerosis (MS). The term "treatment," with respect to a subject, refers to improving at least one symptom of a disease in a subject. Treatment can be curing, ameliorating, or at least partially ameliorating a disease. The term "administer / administering / administration" as used herein refers to administering a compound or a pharma- ceutically acceptable salt or composition of a compound directly to a subject, or administering a prodrug derivative or analog of a compound or a pharma- ceutically acceptable salt or composition of a compound to a subject to form an equivalent amount of an active compound in the subject's body.

[0077] "Patient" refers to an animal, such as a mammal, who has been the object or subject of treatment, observation or experiment. The methods described herein can be used in human treatment and veterinary applications. In some embodiments, the patient is a mammal, in some embodiments the patient is a human, and in some embodiments the patient is selected from a cat and a dog. The term "treatment" refers to the administration of a compound of the present invention to a mammal suffering from a disease by or with the assistance of a competent caregiver, for purposes including (i) prevention of disease, such that clinical symptoms of the disease do not occur, (ii) inhibition of disease, such that clinical symptoms do not develop, and / or (iii) alleviation of disease, such that clinical symptoms are eradicated.

[0078] "Haloalkyl group" refers to a straight or branched chain alkyl group as defined above, in which one or more hydrogen atoms are replaced by halogen. Examples of haloalkyl groups include "fluoroalkyl groups", which include, for example, fluoromethyl, fluoroethyl, fluoropropyl, difluoromethyl, difluoroethyl, difluoropropyl, trifluoromethyl, trifluoroethyl and trifluoropropyl. "Haloalkoxy group" refers to a straight or branched chain alkyl group as defined above, in which one or more hydrogen atoms are replaced by halogen. Examples of haloalkoxy groups include "fluoroalkoxy groups", which include, for example, fluoromethoxy, fluoroethoxy, fluoropropoxy, difluoromethoxy, difluoroethoxy, difluoropropoxy, trifluoromethoxy, trifluoroethoxy and trifluoropropoxy.

[0079] Pharmaceutical Compositions and Administration The present invention provides pharmaceutical compositions comprising the compounds of the present invention and pharma- ceutically acceptable vectors. Pharmaceutical compositions can be formulated for specific routes of administration, such as oral, parenteral and rectal administration. In addition, pharmaceutical compositions of the present invention can be prepared in solid form (including but not limited to capsules, tablets, pills, granules, powders or suppositories) or liquid form (including but not limited to solutions, suspensions or emulsions). Pharmaceutical compositions can be subjected to conventional pharmaceutical operations such as sterilization, and / or can contain conventional inert diluents, lubricants or buffers, and auxiliary materials such as preservatives, stabilizers, wetting agents, emulsifiers and buffers.

[0080] Typically, the pharmaceutical composition is a tablet or gelatin capsule, containing the active ingredient, a) diluents such as lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and / or glycine; b) lubricants, such as silicon dioxide, talc, stearic acid, its magnesium or calcium salts and / or polyethylene glycol, also suitable for tablets; c) binding agents, such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidone, if necessary d) disintegrants such as starch, agar, alginic acid or its sodium salt, or effervescent mixtures, and / or e) Absorbents, colouring agents, flavouring agents and sweetening agents.

[0081] Tablets can be film coated or enteric coated according to methods known in the art. Compositions suitable for oral administration contain an effective amount of a compound of the invention and are in the form of tablets, troches, aqueous or oily emulsions, dispersible powders or granules, emulsions, paraffin or soft capsules, or syrups or elixirs.

[0082] The compositions for oral use are prepared according to any method known in the art for the preparation of pharmaceutical compositions, and such compositions may contain one or more agents selected from sweeteners, flavorings, coloring agents and preservatives to provide a pharma- ceutically elegant and palatable preparation. Tablets may contain the active ingredient mixed with non-toxic pharma- ceutically acceptable excipients, which are suitable for the manufacture of tablets. These excipients are inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate, granulating and disintegrating agents such as corn starch or alginic acid, binding agents such as starch, gelatin or gum arabic, and lubricants such as magnesium stearate, stearic acid or talc. Tablets may be uncoated or coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and provide a longer sustained action. For example, a time-delay material such as glyceryl monostearate or glyceryl distearate may be used. Oral formulations are hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent such as calcium carbonate, calcium phosphate, or kaolin, or soft gelatin capsule media in which the active ingredient is mixed with water or an oil, e.g., peanut oil, liquid paraffin, or olive oil.

[0083] Certain injectable compositions are isotonic aqueous or turbid liquids, and suppositories are preferably prepared from fatty emulsions or suspensions. The compositions can be sterilized and / or contain adjuvants such as preservatives, stabilizers, wetting agents or emulsifiers, solution promoters, salts for regulating osmotic pressure and / or buffers. In addition, they can further contain other substances of therapeutic value. The compositions are prepared according to conventional mixing, granulation or coating methods, respectively, and contain about 0.1-75%, or about 1-50%, of the active ingredient. The composition suitable for transdermal administration comprises an effective amount of the compound of the present invention together with a suitable vector.The vector suitable for transdermal delivery comprises an absorbable pharmacologically acceptable solvent that facilitates passage through the skin of the host.For example, the transdermal device is in the form of a dressing, which comprises a backing member, a reservoir that contains the compound, optionally together with a vector, and optionally a rate-controlling barrier that delivers the compound at a controlled, predetermined rate over a long period of time through the skin of the host, and a device that is placed under the skin.

[0084] Compositions suitable for topical application to the skin and eyes include aqueous solutions, suspensions, ointments, creams, gels, or sprayable formulations for delivery by aerosols, etc. Such topical delivery systems are particularly suitable for dermal applications, such as for the treatment of skin cancer, and for preventative use, such as sunscreens, lotions, sprays, etc. Thus, they are particularly suitable for use in topical formulations well known in the art, including cosmetic formulations. They may include solubilizers, stabilizers, tonicity agents, buffers, and preservatives. As used herein, topical application may also include inhalation or intranasal application. They can be conveniently delivered as a dry powder from a dry powder inhaler (alone, as a mixture, e.g., dry blended with lactose, or mixed with ingredient granules, e.g., mixed with phospholipids) or as an aerosol spray from a processed container, pump, atomizer, or nebulizer, with or without a suitable propellant.

[0085] Dosage forms for topical or transdermal administration of the compounds of the present invention include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active compound can be mixed under sterile conditions with a pharma- ceutically acceptable vector and any preservatives, buffers or propellants which may be required. In addition to the active compounds of this invention, the ointments, pastes, creams and gels may contain excipients such as animal and vegetable fats, oils, waxes, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonite, calcium, talc and zinc oxide, or mixtures thereof. In addition to the compounds of the invention, powders and sprays can contain further excipients, such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays can further contain conventional propellants, such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.

[0086] Transdermal patch has the additional advantage of controlled delivery of the compound of the present invention to the body.The compound can be dissolved or dispersed in a suitable solvent to prepare such a dosage form.Absorption enhancers can also be used to increase the flux of the compound through the skin.The rate of such flux can be controlled by providing a rate-controlling membrane or by dispersing the active compound in a polymer matrix or gel. Ophthalmic formulations, eye ointments, powders, solutions and the like, are also contemplated as being within the scope of the present invention.

[0087] The present invention further provides anhydrous pharmaceutical compositions and dosage forms comprising the compounds of the present invention as active ingredients, since water can accelerate the decomposition of certain compounds.The anhydrous pharmaceutical compositions and dosage forms of the present invention can be prepared using anhydrous or low moisture ingredients and low moisture or low humidity conditions.The anhydrous pharmaceutical composition can be prepared and stored to maintain its anhydrous nature.Thus, the anhydrous composition is packaged using materials known to prevent exposure to water, so that it can be included in a suitable formulary kit.Examples of suitable packaging include, but are not limited to, hermetically sealed foils, plastics, unit dose containers (e.g., vials), blister packs, and strip packs. The present invention further provides pharmaceutical compositions and dosage forms that comprise as an active ingredient one or more agents that reduce the rate of decomposition of the compounds of the present invention. Such agents, which are referred to herein as "stabilizers," include, but are not limited to, antioxidants such as ascorbic acid, pH buffers, or salt buffers.

[0088] An "effective amount" of the disclosed CCR8 inhibitors is an amount that inhibits CCR8 activity in a subject in need of such inhibition, or an amount that, when administered to a subject suffering from a CCR8-mediated disease, ameliorates symptoms of the disease or condition, delays the onset of symptoms, and / or extends lifespan. The exact amount of CCR8 inhibitor administered to a subject can vary depending on the type and severity of the disease or condition, as well as the characteristics of the subject, such as general health, age, sex, weight, and drug tolerance. Dosages also vary depending on the route of administration, such as oral, aerosol, rectal, transdermal, subcutaneous, intravenous, intramuscular, intraperitoneal, intranasal, etc. One of skill in the art can determine appropriate dosages depending on these and other factors. An "effective amount" is usually about 0.01 mg / kg / day to about 100 mg / kg / day, preferably about 0.5 mg / kg / day to about 50 mg / kg / day. A physician, clinician, or veterinarian of ordinary skill can readily determine the effective amount of each active ingredient required to prevent, treat, or inhibit the progress of a disease condition or disorder.

[0089] The compounds of the invention can be administered in vitro in the form of solutions, e.g., aqueous solutions, and in vivo either enterally, parenterally, or conveniently intravenously, e.g., as a suspension or aqueous solution. The therapeutically effective amounts in vivo range from about 0.01 to 100 mg / kg, or from about 0.5 to 50 mg / kg, depending on the route of administration. The compounds of the invention can be administered simultaneously with, before or after, one or more other therapeutic agents. The compounds of the invention can be administered separately by the same or different route of administration, or together with other agents in the same pharmaceutical composition.

[0090] Uses of the compounds and compositions thereof The disclosed compounds are potent inhibitors of CCR8, and therefore are expected to be useful in treating and preventing CCR8-mediated diseases. Diseases for which the disclosed CCR8 inhibitors are described as being effective include, but are not limited to, cancers such as colon cancer, pancreatic cancer, lung cancer and liver cancer, neuropathic pain (caused by diabetes or spinal cord injury), asthma, atopic dermatitis, allergic rhinitis, inflammatory diseases such as systemic allergic reactions or hypersensitivity reactions, drug allergies (e.g., penicillin, cephalosporin), insect bite allergies, and inflammatory dermatitis such as dermatitis, eczema, allergic contact dermatitis, urticaria, atherosclerosis, restenosis, myositis (including polymyositis, dermatomyositis) or opioids. Examples of cancers for which the disclosed compounds, pharmaceutical compositions and methods are particularly effective include leukemia (including the leukemic stages of chronic lymphocytic leukemia, chronic myeloid leukemia, acute myeloid leukemia, acute lymphoblastic leukemia and lymphoma) and solid tumors (including breast cancer, gastric cancer, ovarian cancer, pancreatic cancer, liver cancer, etc.).

[0091] General Synthesis Exemplary embodiments of the compounds of the present invention can be synthesized using the general reaction scheme described below. It will be clear from the description herein that the general scheme can be modified by replacing the starting materials with others of similar structure to produce different corresponding products. The synthetic description provides many examples of how the starting materials were modified to obtain the corresponding products. The starting materials are usually obtained from commercial sources or synthesized using published methods. The compounds of the present invention can be prepared starting from commercially available starting materials using general synthetic techniques and procedures known to those skilled in the art. Below, a suitable scheme for the preparation of such compounds is outlined. Further examples can be found in the specific examples detailed below.

[0092] General Method A JPEG0007653166000149.jpg34147

[0093] General Method B JPEG0007653166000150.jpg34145

[0094] General Method C JPEG0007653166000151.jpg28145

[0095] General Method D JPEG0007653166000152.jpg36144JPEG0007653166000153.jpg36144

[0096] As mentioned above, A is a 6-, 9-, 10- or 11-membered carbocyclic or heterocyclic ring; where A is halo, a C1-6 alkyl group, a C1-6 alkyl group substituted with halo, C 1-6 Alkoxy group, C 3-6 Cycloalkyl groups, -COOH, -NH2, -CN, -NHCO(C 1-6 alkyl), -NH(C 1-6 alkyl), -N(C 1-6 alkyl)2, -CONH2, -COO(C 1-6 alkyl), -OCO(C 1-6 alkyl), -CONH(C 1-6 alkyl), CON(C 1-6 and each of the aryl, aryl, and alkyl groups may be substituted by 0, 1, 2, or 3 substituents independently selected from m is 0 or 1; p is 0 or 1; t is 0, 1, 2 or 3; n is 0 or 1, R5 is O or NH; R1 is H or C 1-6 is an alkyl group, R2 and R3 are each independently H, C 1-6 Alkyl groups, -CH2OH, -COOCH3, C 3-6 is selected from a cycloalkyl group, a phenyl group, a heterocyclic ring, or a heteroaryl ring;

[0097] R4 is C 1-6 B3 is a 5- or 6-membered carbocyclic ring having a saturated or unsaturated hydrocarbon; where the carbon atoms of the phenyl group can be substituted by 0, 1 or 2 N, and can be substituted with 4, 5, 6, 7 or 8-membered cycloalkyl groups, 5-membered heteroaryl rings, bridged (C 5-12 ) the carbon atoms of the cycloalkyl group, B1 or B2, can be substituted by 0, 1, or 2 heteroatoms independently selected from N, O, or S, and the carbon atoms of B3 can be substituted by 0, 1, 2, or 3 heteroatoms independently selected from N, O, or S;

[0098] Phenyl groups, 4-, 5-, 6-, 7- or 8-membered cycloalkyl groups, bridged (C 5-12 ) a cycloalkyl group, JPEG0007653166000154.jpg44160 is a group consisting of halo, C1-6 alkyl group, C1-6 alkyl group substituted with halo, C 3-8 Cycloalkyl groups, -O(C 1-6 alkyl), -OH, -NH2, -N(C 1-6 alkyl)2, -NHCO(C 1-6 alkyl), -NHBoc, -COOH, -COO(C 1-6 alkyl), -COOC(C 1-6 Alkyl)3, -CO(C 1-6 alkyl), -CH2COO(C 1-6 alkyl), -CO(CH2)qN(C 1-6 alkyl)2, -COCH2NH2, -CH2CON(C 1-6 alkyl)2, -CH2CONH(C 1-6 alkyl), -(C 1-6 alkyl)OH, -COCH2NHBoc, -COCH(CH3)(NHBoc), -(C 1-6 Alkyl)N(C 1-6 alkyl)2, -CH2CF3, -(C 1-6 Alkyl)O(C 1-6 alkyl), α-amino acid group, Optionally substituted by 0, 1, 2 or 3 substituents independently selected from:

[0099] Here, C 3-8 The carbon atoms of the cycloalkyl group may be substituted by 0, 1 or 2 heteroatoms independently selected from O, N and S; or R4 is JPEG0007653166000156.jpg34160, where q is 0 or 1, Y1 is CH or N; Y2 is CH2, O, or NR6, and R6 is H or C 1-6 is an alkyl group, Y3 is -CH2OH, -COO(C 1-6 alkyl), -(C 1-6 Alkyl)N(C 1-6 alkyl)2, -N(C 1-6 alkyl)2,

[0100] X is JPEG0007653166000157.jpg109160-NHCOC(C 1-6 alkyl)3;

[0101] where r is 0, 1, 2, 3, 4, and R7 is H or C 1-6 is an alkyl group, JPEG0007653166000158.jpg34160 is -OH,C 1-6 Alkyl group, C 3-6 Cycloalkyl groups, halo, halo-substituted C1-6 alkyl groups, C 1-6 optionally substituted with 0, 1, 2 or 3 substituents independently selected from alkoxy groups; D is a 4-, 5-, 6-, 7- or 8-membered cycloalkyl group other than a phenyl group, or a bridged (C 5-12) a cycloalkyl group, wherein the carbon atoms of D can be replaced by 0, 1 or 2 heteroatoms independently selected from N or O, and D is C 1-3 Alkyl group, halo, -COOH, C 1-6 optionally substituted with 0, 1 or 2 substituents independently selected from alkoxy groups; E1 is C 3-7 is a cycloalkyl group, and E2 is CF3 or C 3-6 is a cycloalkyl group,

[0102] wherein the carbon atom of E2 can be substituted by 0 or 1 heteroatom independently selected from N, O or S; JPEG0007653166000159.jpg38160 carbon atoms in the ring may be replaced by 0, 1 or 2 heteroatoms independently selected from N, O or S; The E group can undergo a series of reactions to give X, The R8 group can undergo a series of reactions to give R4. or a pharma- ceutically acceptable salt, ester, isotope, stereoisomer, tautomer, solvate, prodrug, or combination thereof. EXAMPLES

[0103] Preparation and Examples The present invention can be better understood based on the following examples. However, those skilled in the art can easily understand that the contents described in the examples are only for the purpose of illustrating the present invention, and are not intended to limit the present invention described in detail in the claims.

[0104] Unless otherwise stated, the compounds of the present invention can be prepared starting from commercially available starting materials and utilizing common synthetic techniques and procedures known to those skilled in the art. Chromatographic consumables and devices can be purchased from companies such as AnaLogix, Inc, Burlington, WI, Analytical Sales and Services, Inc., Pompton Plains, NJ, TeledyneIsco, Lincoln, NE, VWR International, Bridgeport, NJ, and Rainin Instrument Company, Woburn, MA. Chemicals and reagents can be purchased from companies such as Aldrich, Argonaut Technologies, VWR and Lancaster, Invitrogen, Sigma, Promega, Solarbio, Cisbio, Signalchem, MCE, consumables can be purchased from companies such as Corning, Labcyte, Greiner, Nunc, and equipment can be purchased from companies such as Labcyte, PerkinElmer, Eppendorf, ThermoFisher, etc.

[0105] Preparation of materials Synthesis of general intermediates C1 & C2 JPEG0007653166000160.jpg3658

[0106] Compound 1 (C1) is prepared according to the reference Journal of Medicinal Chemistry (2007), 50(3), 566-584. C1 (1 g, 2.78 mmol) is dissolved in acetone (20 mL), then aqueous ammonia (2.2 mL, 13.90 mmol) is added at 0° C., and the mixture is stirred for 3 h. After precipitation, the solid is collected by filtration and then dried to give the desired product compound 2 (C2) (0.8 g) as a white solid. LC-MS(ESI):m / z 341.1[M+H] + ,

[0107] Working Example Example 1 JPEG0007653166000161.jpg28144

[0108] Pd2(dba)3 (0.39 g, 0.43 mmol), BINAP (0.54 g, 0.86 mmol) and sodium 2-methylpropan-2-olate (2.90 g, 30.17 mmol) are added to the solution of compound 1-2 (2.60 g, 12.93 mmol) and compound 1-1 (2 g, 8.62 mmol) in anhydrous toluene. The reaction mixture is thoroughly degassed using a stream of nitrogen. The resulting mixture is stirred at 110 °C for 16 h and then concentrated to give the crude product. The residue is purified by flash silica gel chromatography (45% ethyl acetate in petroleum ether) to give compound 1-3 (1.1 g, 2.44 mmol, 28.3%) as a yellow oil. LC-MS(ESI):m / z 280.3[M+H] + .

[0109] Under a stream of nitrogen, palladium on carbon (Pd / C, 1.1 g) is added to a solution of compound 1-3 (1.1 g, 2.44 mmol) in MeOH (50 mL). The suspension is degassed under vacuum and flushed with hydrogen gas three times. The mixture is stirred under hydrogen gas (15 psi) at 30° C. for 16 h. The mixture is filtered, and the filtrate is then concentrated to give the crude product. The residue is purified by flash column chromatography (50% tetrahydrofuran in petroleum ether) to give the desired product 1-4 (0.6 g, 2.12 mmol) as a yellow oil. LC-MS(ESI):m / z 250.3[M+H] + .

[0110] A solution of compound C1 (90 mg, 0.25 mmol) and product 1-4 (57 mg, 0.18 mmol) in pyridine (0.2 mL) is stirred at 30 °C for 16 h. The mixture is concentrated to give the crude product. The residue is purified by preparative HPLC (YMC-Actus Triart C18 150 × 30 mm × 5 um, mobile phase: [water (0.05% ammonia v / v)-acetonitrile], B%: 55%-75%, 10 min) to give the desired product 1 (40 mg, 0.07 mmol, 39.3%) as a white solid.

[0111] The following compounds are synthesized according to the steps described in Example 1. JPEG0007653166000162.jpg237160JPEG0007653166000163.jpg246160JPEG0007653166000164.jpg246160JPEG0007653166000165.jpg99160

[0112] Example 2 JPEG0007653166000166.jpg43107

[0113] Compound 2-1 (16.3 mg, 0.081 mmol), DMAP (26.9 mg, 0.22 mmol) and EDC HCl (42.2 mg, 0.22 mmol) are added sequentially to a suspension of compound C2 (25 mg, 0.073 mmol) dissolved in DCM (1 mL) at 25 °C. The reaction mixture is stirred for 16 h at 25 °C. 2 mL of dilute HCl (1 N) is added to the reaction mixture, extracted with DCM (1 mL x 3), and the combined organic layers are concentrated under reduced pressure to obtain a residue. The residue is purified by preparative HPLC (column: Xtimate C18 150 x 40 mm x 10 um [water (0.225% formic acid)-acetonitrile] B%: 40%-80%, 10 min) to obtain a white solid product compound 5 (4.30 mg, 0.01 mmol, 11.2%).

[0114] The following compounds are synthesized according to the steps described in Example 2. JPEG0007653166000167.jpg239160JPEG0007653166000168.jpg246160JPEG0007653166000169.jpg246160 JPEG0007653166000170.jpg227160JPEG0007653166000171.jpg235160JPEG0007653166000172.jpg112160

[0115] Example 3 JPEG0007653166000173.jpg42146JPEG0007653166000174.jpg46117

[0116] ] At 25°C, C1 (512 mg, 1.42 mmol) is added to a solution of compound 14-1 (100 mg, 0.59 mmol) in pyridine (1 mL, 12.4 mmol). The reaction mixture is stirred for 16 h at 25°C. 3 mL of water is added to the reaction mixture, and it is extracted with DCM (1 mL x 3), and the combined organic layers are concentrated under reduced pressure to obtain a residue. The residue is purified by preparative HPLC (YMC Triart C18 150 x 25 mm x 5 um [water (0.225% formic acid)-acetonitrile] B%: 60%-80%, 10 min) to obtain white solid compound 24 (121 mg, 0.22 mmol, 20.0%).

[0117] Compound 24 (20 mg, 36.25 μmol) is added in one portion to HCl / dioxane (1 mL, 4 mol / L) at 25° C. The reaction mixture is stirred for 16 h at 25° C. The reaction mixture is concentrated under reduced pressure to give a residue. The residue is purified by preparative HPLC (column: Diamonsil C18 150×30 mm×5 um [water (0.225% formic acid)-acetonitrile] B%: 27%-67%, 10 min) to give compound 17 (0.68 mg, 3.8%) as a white solid.

[0118] DIEA (17μL, 132μmol)) and acetyl chloride (3.5μL, 48.7μmol) are added sequentially to a solution of compound 17 (20mg, 44.3μmol) dissolved in DCM (3mL) at 0℃. The reaction mixture is heated to 25℃ and stirred at 25℃ for 16h. 3mL of water is added to the reaction mixture and extracted with DCM (1mL×3). The combined organic layer is concentrated under reduced pressure. The residue is purified by preparative HPLC (column: Phenomenex Gemini C18 250×50mm×10um [water (0.225% formic acid)-acetonitrile] B%: 35%-55%, 10min) to give white solid compound 18A (1.03mg, 4.7%) and I compound 18B (1.18mg, 5.4%).

[0119] Potassium carbonate (18.4 mg, 133 μmol) and NaI (0.7 mg, 4.4 μmol) are added successively to a solution of compound 17 (20 mg, 44.3 μmol) dissolved in acetonitrile (2 mL) at 25° C., and the reaction mixture is stirred for 16 h at 25° C. 3 mL of water is added to the reaction mixture, and it is extracted with DCM (1 mL×3), and the combined organic layer is concentrated under reduced pressure. The residue is purified by preparative HPLC (Phenomenex Gemini C18 250×50 mm×10 um [water (0.05% ammonia water v / v)-acetonitrile] B%: 48%-68%, 10 min) to give white solid compound 19A (1.12 mg, 2.1 μmol, 4.8%) and white solid compound 18B (1.20 mg, 2.3 μmol, 5.2%).

[0120] The following compounds are synthesized according to the steps described in Example 3. JPEG0007653166000175.jpg224160JPEG0007653166000176.jpg246160JPEG0007653166000177.jpg246160JPEG0007653166000178.jpg246160JPEG0007653166000179.jpg246160JPEG0007653166000180.jpg220160JPEG0007653166000181.jpg210160JPEG0007653166000182.jpg245160JPEG0007653166000183.jpg241160JPEG0007653166000184.jpg245160JPEG0007653166000185.jpg245160JPEG0007653166000186.jpg245160JPEG0007653166000187.jpg241160JPEG0007653166000188.jpg245160JPEG0007653166000189.jpg238160JPEG0007653166000190.jpg245160JPEG0007653166000191.jpg245160JPEG0007653166000192.jpg245160JPEG0007653166000193.jpg241160JPEG0007653166000194.jpg245160JPEG0007653166000195.jpg245160JPEG0007653166000196.jpg234160JPEG0007653166000197.jpg245160JPEG0007653166000198.jpg245160JPEG0007653166000199.jpg245160JPEG0007653166000200.jpg245160JPEG0007653166000201.jpg245160JPEG0007653166000202.jpg245160JPEG0007653166000203.jpg241160JPEG0007653166000204.jpg245160JPEG0007653166000205.jpg234160JPEG0007653166000206.jpg245160JPEG0007653166000207.jpg245160JPEG0007653166000208.jpg245160JPEG0007653166000209.jpg241160JPEG0007653166000210.jpg245160JPEG00076531660 00211.jpg241160JPEG0007653166000212.jpg245160JPEG0007653166000213.jpg241160JPEG0007653166000214.jpg245160JPEG000765 3166000215.jpg224160JPEG0007653166000216.jpg243160JPEG0007653166000217.jpg245160JPEG0007653166000218.jpg245160JPEG0 007653166000219.jpg245160JPEG0007653166000220.jpg246160JPEG0007653166000221.jpg246160JPEG0007653166000222.jpg29160.

[0121] Example 4 JPEG0007653166000223.jpg82145

[0122] DIEA (5.4 mL, 32.4 mmol) is added to a solution of compound 4-1 (1 g, 5.4 mmol) and compound 1-2 (3.11 g, 10.8 mmol) dissolved in DMSO (5 mL) at 25° C. The reaction mixture is heated to 120° C. and stirred for 16 h. The reaction mixture is then cooled to 25° C. and 25 mL of water is added. The reaction mixture is acidified to pH 2 with dilute HCl and extracted with DCM (20 mL×6). The combined organic layer is dried over Na2SO4. After filtration, the filtrate is concentrated under reduced pressure to give the remaining compound 4-2 (870 mg, 2.97 mmol, 54.9%) as a brown oil. The residue is used in the next step without further purification. LC-MS: (ESI+) m / z 294.2 [M+H] + .

[0123] A solution of NH3H2O ​​(2 mL) and Pd / C (50 mg, 0.48 mmol) are added sequentially to a solution of compound 4-2 (100 mg, 3.4 mmol) in methanol (20 mL) at 25° C. The reaction mixture is stirred under hydrogen gas for 16 h at 25° C. The reaction mixture is filtered and the filtrate is concentrated under reduced pressure to give a colorless oily residue of compound 4-3 (144 mg, 0.27 mmol, 80.2%). The residue is used in the next step without further purification. LC-MS: (ESI+) m / z 264.2 [M+H] + .

[0124] Compound C1 (68.3 mg, 0.19 mmol) is added to a solution of compound 4-3 (100 mg, 0.19 mmol) dissolved in pyridine (1 mL, 12.4 mmol) under 0°C. The reaction mixture is heated to 25°C and stirred for 16 hours. 2 mL of dilute HCl (0.5N) is added to the reaction mixture and extracted with DCM (1 mL x 3). The combined organic layer is dried over Na2SO4. After filtration, the filtrate is concentrated under reduced pressure to obtain a residue. The residue is purified by preparative HPLC (column: YMC Triart C18 250 x 50 mm x 7 um [water (0.05% ammonia water v / v)-acetonitrile] B%: 29%-69%, 10 min) to obtain white solid compound 91 (5.98 mg, 0.01 mmol, 5.4%).

[0125] The following compounds are synthesized according to the steps described in Example 4. JPEG0007653166000224.jpg236160JPEG0007653166000225.jpg246160JPEG0007653166000226.jpg58160

[0126] Example 5 JPEG0007653166000227.jpg58141

[0127] DIEA (4.8mL, 29.12mmol) is added to a solution of compound 5-1 (0.6mL, 4.85mmol) and compound 1-2 (2.10g, 7.28mmol) dissolved in DMSO (10mL) at 25°C. The reaction mixture is heated to 120°C and stirred for 16 hours. The reaction mixture is then cooled to 25°C, 50mL of water is added to the reaction mixture, and extracted with ethyl acetate (20mL x 3). The combined organic layer is dried over Na2SO4. After filtration, the filtrate is concentrated under reduced pressure to give a residue of brown oily compound 5-2 (1.3g, 4.14mmol, 85.2%). The residue is used in the next step without further purification. LC-MS: (ESI+) m / z 314.1 [M+H] + .

[0128] Under nitrogen gas at 25°C, T-BuONa (183.3mg, 1.91mmol), Xantphos (11.0mg, 0.019mmol) are first added to a solution of compound 5-2 (150mg, 0.48mmol) and diphenylmethylamine (0.20mL, 1.19mmol) in toluene (4.5mL), followed by Pd2(dba)3 (8.7mg, 0.01mmol). The reaction mixture is heated to 100°C and stirred for 16 hours. The reaction mixture is then cooled to 25°C, 15mL of water is added, and extracted with ethyl acetate (5mL x 3). The combined organic layer is dried over Na2SO4. After filtration, the filtrate is concentrated under reduced pressure to give a residue of brown oily compound 5-3 (230mg, 0.28mmol, 58.1%). The residue is used in the next step without further purification. LC-MS:(ESI+)m / z 251.3[M+H-169] + .

[0129] HCl / dioxane (1.1 mL, 4.4 mmol) is added to a solution of compound 5-3 (90 mg, 0.22 mmol) in DCM (2 mL) at 25° C. The reaction mixture is stirred for 16 h at 25° C. The reaction mixture is then concentrated under reduced pressure to give a residue of compound 5-4 (55 mg, 0.13 mmol, 61.0%). The residue is used in the next step without further purification. LC-MS: (ESI+) m / z 280.2 [M+H] + .

[0130] Compound C1 (69.9 mg, 1.94 mmol) is added to a solution of compound 5-4 (54 mg, 0.13 mmol) in pyridine (4 mL, 49.55 mmol) under 0°C. The reaction mixture is heated to 25°C and stirred for 16 hours. 5 mL of water and 1 mL of dilute HCl (1N) are added to the reaction mixture and extracted with DCM (2 mL x 3). The combined organic layer is dried over Na2SO4. After filtration, the filtrate is concentrated under reduced pressure to obtain a residue. The residue is purified by HPLC (column: YMC-Actus Triart C18 150 x 30 mm x 5 um [water (0.05% ammonia water v / v)-acetonitrile] B%: 49%-69%, 10 min) to obtain white solid compound 94 (11.1 mg, 0.02 mmol, 14.9%).

[0131] The following compounds are synthesized according to the steps described in Example 5. JPEG0007653166000228.jpg188160

[0132] Example 6 JPEG0007653166000229.jpg70144

[0133] Acetyl acetate (118 μL, 1.26 mmol) is added to a solution of compound 6-1 (200 mg, 1.05 mmol) in pyridine (2 mL) at 10° C. The mixture is stirred for 16 h at 30° C. and concentrated to give crude product I, compound 6-2 (200 mg, crude), which is used in the next step without purification. LC-MS(ESI):m / z 232.0[M+H] + .

[0134] Under a nitrogen stream, the starting materials Pd2(dba)3 (78.9 mg, 0.086 mmol), Davephos(2-(Di-Tert-Butylphosphino)Biphenyl) (33.9 mg, 0.086 mmol) and sodium 2-methylpropan-2-olate (347.9 mg, 3.62 mmol) are added sequentially to a solution of compound 1-2 (375 mg, 1.29 mmol) and compound 6-2 (200 mg, 0.86 mmol) dissolved in anhydrous dioxane (2 mL). The resulting mixture is stirred at 110 °C for 16 h. The mixture is filtered and the filtrate is concentrated to give the crude product. The residue was purified by preparative HPLC (column: YMC-Pack CN150×30 mm×5 um, mobile phase: heptane-ethanol; B%: 0%-70%, 14 min) to give yellow solid compound 6-3 (120 mg, 0.39 mmol, 37.4%). LC-MS(ESI):m / z 280.2[M+H] + .

[0135] A solution of compound 6-3 (120 mg, 0.43 mmol) in HCl solution (5 M, 1 mL, 5.0 mmol) is heated at 100° C. for 16 h. The mixture is diluted with DCM (5 mL) and basified with saturated NaHCO3 to pH=9. The aqueous layer is separated and extracted with DCM (5 mL×1). The combined organic phase is washed with brine, dried over Na2SO4, and concentrated in vacuo to give compound 6-4 (70 mg, 0.29 mmol, 68.7%) as a brown oil. LC-MS(ESI):m / z 238.2[M+H] + .

[0136] A solution of compound C1 (106 mg, 0.30 mmol) and compound 6-4 (70 mg, 0.30 mmol) in pyridine (1 mL) is stirred for 16 hours at 30° C. The mixture is concentrated and purified by preparative HPLC (column: Phenomenex Gemini C18 250×50 mm×10 um, mobile phase: water (0.05% aqueous ammonia v / v)-acetonitrile, B%: 50%-70%, 10 min) to obtain the desired product, white solid compound 95 (38.5 mg, 0.07 mmol, 23.0%).

[0137] The following compounds are synthesized according to the steps described in Example 6. JPEG0007653166000230.jpg188160

[0138] Example 7 JPEG0007653166000231.jpg97144

[0139] A solution of 2-[(2-hydroxyethyl)amino]-1-ol (7 mL, 73 mmol) in tetrahydrofuran (10 mL) is cooled to 0° C., and then a solution of compound 7-1 (2 g, 9.1 mmol) in tetrahydrofuran (10 mL) is slowly added dropwise. The mixture is stirred at 0° C. for 1 h, and then at 25° C. for 16 h. The mixture is concentrated and diluted with DCM (30 mL). The mixture is washed with H2O (20 mL×3). The combined aqueous layer is extracted with DCM (20 mL×3). The combined organic layer is dried over MgSO4, filtered, and concentrated to give a yellow oily crude product compound 7-2 (0.96 g, 2.99 mmol, 32.9%). LC-MS(ESI):m / z 289.1,291.0[M+H] + .

[0140] Compound 7-2 (880 mg, 2.7 mmol) is added to a solution of KOH (154 mg, 2.7 mmol) in HO (2.7 mL), and the resulting mixture is stirred at 100° C. for 2 h. The reaction mixture is cooled to room temperature, and then a yellow solid is precipitated. After filtration, the solid is washed with HO (1 mL×2) and dried under vacuum to obtain compound 7-3 (210 mg, 0.83 mmol, 60.4%) as a yellow solid. LC-MS(ESI):m / z 253.1[M+H] + .

[0141] Dess-Martin reagent (126 mg, 0.30 mmol) is added to a solution of compound 7-3 (50 mg, 0.20 mmol) in DCM (2 mL). The mixture is stirred at 25° C. for 2 h. The mixture is diluted with saturated NaHCO3 (6 mL) and extracted with DCM (2 mL×3). The combined organic layers are dried over MgSO4, filtered and concentrated to give compound 7-4 (55 mg, 0.18 mmol, 90.9%) as a white oily crude product. LC-MS(ESI):m / z 251.1[M+H] + .

[0142] Triethylamine (125 μL, 0.90 mmol) and NaBH(OAc)3 (228 mg, 1.08 mmol) are added to a solution of compound 7-4 (55 mg, 0.18 mmol) and dimethylamine hydrochloride (44 mg, 0.54 mmol) in DCM (2 mL). The mixture is stirred at 25 °C for 16 h. The mixture is diluted with saturated NaHCO3 (6 mL) solution and extracted with DCM (2 mL x 4). The combined organic layer is dried over MgSO4, filtered and concentrated to give compound 7-5 (52 mg, 0.15 mmol, 81.7%) as a yellow oily crude product. LC-MS(ESI):m / z 279.9[M+H] + .

[0143] A solution of compound 7-5 (52 mg, 0.15 mmol) and DIEA (244 μL, 1.47 mmol) in DCM (0.7 mL) is cooled to 0° C. Then a solution of trichlorosilane (127 μL, 0.74 mmol) in DCM (0.7 mL) is added. The mixture is stirred at 25° C. for 11 h. The mixture is diluted with saturated NaHCO3 (6 mL) solution. Extraction is performed with DCM (2 mL×4). The combined organic layer is dried over MgSO4, filtered and concentrated to give compound 7-6 (25 mg, 0.10 mmol, 68.2%) as a yellow oily crude product. LC-MS(ESI): m / z 250.1[M+H] + .

[0144] Compound C1 (28.9 mg, 0.08 mmol) was added to a mixture of compound 7-6 (25 mg, 0.10 mmol) dissolved in pyridine (1 mL). The mixture was concentrated and the crude product was purified by preparative HPLC (column: Boston Prime C18 150×30 mm×5 um, mobile phase: [water (0.05% ammonia v / v)-acetonitrile], B%: 37%-60%, 10 min) to obtain pure compound 95 (2.4 mg, 4.2 μmol, 4.2%) as a white solid.

[0145] The following compounds are synthesized according to the steps described in Example 7. JPEG0007653166000232.jpg160160

[0146] Example 8 JPEG0007653166000233.jpg62119

[0147] A solution of titanium(IV) isopropylate (2.97 g, 10.46 mmol) and 33% methylamine in ethanol (6.2 mL, 52.3 mmol) is added to a solution of compound 8-1 (1.0 g, 5.23 mmol) in ethanol (5 mL). The mixture is stirred at 25° C. for 12 h. NaBH4 (353.6 mg, 10.46 mmol) is then added and stirred at 25° C. for 3 h. The mixture is concentrated. Water (15 mL) and DCM (20 mL) are then added. After filtration, the filtrate is extracted with DCM (20 mL×3). The combined organic phase is dried over Na2SO4 and concentrated to give compound 8-2 (1.5 g, 7.27 mmol, crude) as a brown oil. LC-MS(ESI):m / z 207.2[M+H] + .

[0148] Propylphosphonic acid cyclic anhydride (T3P, 50% in ethyl acetate, 4.16 g, 13.09 mmol) is added to a solution of 2-(dimethylamino)acetic acid (810 mg, 7.86 mmol) and triethylamine (1.8 mL, 13.09 mmol) in MeCN (30 mL). The mixture is stirred at 25° C. for 1 h. Compound 8-2 (1.08 g, 5.24 mmol) is added and stirred at 25° C. for 12 h and at 50° C. for 3 h. The mixture is concentrated and water (20 mL) is added, followed by extraction with ethyl acetate (30 mL×3). The combined organic phase is dried over Na2SO4, the filtrate is filtered and concentrated. The crude product is purified by flash silica gel chromatography (ISCO, 12 g SepaFlash Silica Flash Column, 0-50% tetrahydrofuran / ethyl acetate gradient eluent: 30 mL / min) to give compound 8-3 (600 mg, 2.06 mmol, 39.3%) as a brown oil. LC-MS(ESI):m / z 292.0[M+H] + .

[0149] Pd / C (200 mg) is added to a solution of compound 8-3 (150 mg, 0.52 mmol) in ethyl acetate (20 mL). The mixture is stirred for 12 h under 45 Psi hydrogen gas at 25° C. The mixture is filtered, washed with methanol (30 mL), and the filtrate is concentrated to give compound 8-4 (125 mg, 0.48 mmol, 92.9%) as a brown oil. LC-MS(ESI):m / z 262.2[M+H] + .

[0150] Compound C1 (82.6 mg, 0.23 mmol) is added to a solution of compound 8-4 (50 mg, 0.19 mmol) in pyridine (1 mL) at 0° C. The mixture is stirred for 12 h at 25° C. and concentrated. The residue is purified by preparative HPLC (column: Xtimate C18 150×40 mm×10 um, mobile phase: water (0.05% ammonia water v / v)-acetonitrile, B%: 43%-83%, 10 min) to obtain white solid compound 97 (70 mg, 0.12 mmol, 63.6%).

[0151] The following compounds are synthesized according to the steps described in Example 8. JPEG0007653166000234.jpg236160JPEG0007653166000235.jpg122160

[0152] Example 9 JPEG0007653166000236.jpg43146

[0153] LiAlH4 (12 mg, 0.32 mmol) is added to a solution of compound 8-4 (75 mg, 0.29 mmol) in tetrahydrofuran (1 mL) at 0°C. The mixture is stirred for 1 h at 25°C. Water (0.1 mL) is added to quench the reaction. The mixture is filtered and the filtrate is concentrated to give the crude product compound 8-5 (70 mg, 0.28 mmol, 98.6%) as a brown oil. LC-MS(ESI):m / z 248.1[M+H] + .

[0154] Compound C1 (122.2 mg, 0.34 mmol) is added to a solution of compound 8-5 (70 mg, 0.28 mmol) in pyridine (1 mL) at 0° C. The mixture is stirred for 12 h at 25° C. and concentrated. The residue is purified by preparative HPLC (column: Xtimate C18 150×40 mm×10 um, mobile phase: water (0.05% ammonia water v / v)-acetonitrile, B%: 60%-100%, 10 min) to obtain white solid compound 99 (21 mg, 0.04 mmol, 13.1%).

[0155] The following compounds are synthesized according to the steps described in Example 9. JPEG0007653166000237.jpg167160

[0156] Example 10 JPEG0007653166000238.jpg68145

[0157] To a solution of compound 10-4 (70 mg, 0.31 mmol) and compound 10-1 (35 mg, 0.31 mmol) in DCM (5 mL), DIEA (0.15 mL, 0.92 mmol), EDC HCl (118 mg, 0.62 mmol) and HOBt (83 mg, 0.62 mmol) are added in sequence. The reaction mixture is stirred at 25° C. for 16 h. 3 mL of water is added to the reaction mixture and extracted with DCM (1 mL×3). The combined organic layer is concentrated under reduced pressure to give a colorless oily residue of compound 10-2 (100 mg, 0.22 mmol, 70.3%). The residue is used in the next step without further purification. LC-MS: (ESI+) m / z 324.3 [M+H] + .

[0158] Compound 10-2 (100 mg, 0.22 mmol) is added to HCl / dioxane (3 mL, 4 mol / L) at 25° C., the reaction mixture is stirred for 16 h at 25° C., and concentrated under reduced pressure to give compound 10-3 (100 mg, 0.19 mmol, 88.9%) as a white solid, which is used in the next step without further purification.

[0159] Add 2-naphthalenesulfonyl chloride (55.8 mg, 0.25 mmol) to a solution of compound 10-3 (100 mg, 0.22 mmol) in pyridine (1 mL, 12.4 mmol) at 25 °C. Then stir the reaction mixture for 16 h at 25 °C. The reaction mixture is a yellow solution. Add 1 mL of water to the reaction mixture to quench the reaction. Concentrate the reaction mixture under reduced pressure to obtain a residue. Purify the residue by preparative HPLC (column: YMC-Actus Triart C18 150 × 30 mm × 7 um [water (0.05% ammonia water v / v)-acetonitrile] B%: 30%-70%, 10 min) to obtain white solid compound 100 (9.36 mg, 0.02 mmol, 8.8%).

[0160] The following compounds are synthesized according to the steps described in Example 10. JPEG0007653166000239.jpg135160

[0161] Example 11 JPEG0007653166000240.jpg146142

[0162] Compound 11-1 (1.5 g, 6.80 mmol) is added to a solution of compound 11-2 (1.63 g, 8.16 mmol) and triethylamine (1.9 mL, 13.60 mmol) in DCM (40 mL) at 0° C. The mixture is slowly warmed to 25° C. and stirred for 16 h. The mixture is washed with H2O (20 mL) and then 1N HCl (30 mL) is added. The organic solvent is removed and precipitated to give a white solid. The solid is filtered off, washed with H2O (5 mL×2), and dried under vacuum to give compound 11-3 (1.92 g, 4.99 mmol, 73.6%) as a white solid. LC-MS(ESI):m / z 407.0[M+Na] + ,329.0[M-56+H] + .

[0163] To a mixture of compound 11-3 (600 mg, 1.56 mmol) and compound 11-4 (169 mg, 1.56 mmol) in dioxane (2 mL), add T3P (2.78 mL, 4.68 mmol) and DIEA (1.03 mL, 6.24 mmol). Stir the mixture at 100 °C for 16 h. Quench the mixture with saturated NaHCO3 (10 mL) and remove the organic solvent under reduced pressure. Filter off the precipitated solid, wash with water (10 mL x 2), and dry under vacuum to obtain crude compound 101 (300 mg, 0.59 mmol, 38.0%) as a yellow solid. The crude product (40 mg) was purified by preparative HPLC (column: Phenomenex Gemini-NX 150 x 30 mm x 5 um, mobile phase: water (0.05% aqueous ammonia v / v)-acetonitrile, B%: 45%-65%, 10 min) to give compound 102 (7.2 mg, 0.02 mmol, 1.0%) as a white solid.

[0164] To a mixture of compound 102 (300 mg, 0.66 mmol) in DCM (2 mL), add HCl / dioxane (0.6 mL, 2.4 mmol). Stir the mixture at 25 °C for 16 h. Concentrate the mixture to give compound 105A (450 mg, 0.63 mmol, 95.9%) as a brown solid crude product. Purify the crude product (50 mg) by preparative HPLC (column: Phenomenex Gemini-NX 150 x 30 mm x 5 um, mobile phase: water (0.05% ammonia v / v)-acetonitrile, B%: 30%-50%, 10 min) to give compound 105A (5.82 mg, 0.01 mmol, 2.3%) as a white solid product.

[0165] To a solution of compound 105A (100 mg, 0.15 mmol), triethylamine (0.2 mL, 1.54 mmol) in DCM (1.5 mL), add 4-methylpiperazine-1-carbonyl chloride (25 mg, 0.15 mmol), then stir the mixture at 30 °C for 16 h. Wash the mixture with saturated NaHCO3 (10 mL), dry over Na2SO4, filter and concentrate the filtrate. Purify the residue by preparative HPLC (column: Phenomenex Gemini-NX 150 x 30 mm x 5 um, mobile phase: water (0.05% aqueous ammonia v / v)-acetonitrile, B%: 28%-48%, 10 min) to obtain the desired product as a white solid compound 103 (19.5 mg, 0.04 mmol, 27.0%).

[0166] To a solution of compound 105A (100 mg, 0.15 mmol), triethylamine (0.2 mL, 1.54 mmol) in DCM (1.5 mL), 4-methylpiperazine-1-carbonyl chloride (25 mg, 0.15 mmol) is added, and then the mixture is stirred for 16 h at 30° C. The mixture is washed with saturated NaHCO3 (10 mL×1), dried over Na2SO4, filtered and concentrated to give crude yellow oily compound 11-5 (80 mg, 0.14 mmol, 90.0%). LC-MS(ESI):m / z 528.3[M+H] + ,

[0167] To a mixture of compound 11-5 (80 mg, 0.15 mmol) in DCM (1 mL), add HCl / dioxane (0.38 mL). Stir the mixture at 30 °C for 16 h. Concentrate the mixture to obtain the crude product. Purify the residue by preparative HPLC (column: Phenomenex Gemini C18 250 x 50 mm x 10 um, mobile phase: water (0.05% ammonia v / v)-acetonitrile, B%: 25%-45%, 10 min, column: Phenomenex Gemini C18 250 x 50 mm x 10 um, mobile phase: water (0.225% formic acid)-acetonitrile, B%: 5%-25%, 10 min) to obtain the desired product, white solid compound 104 (9 mg, 0.02 mmol).

[0168] The following compounds are synthesized according to the steps described in Example 11. JPEG0007653166000241.jpg233160JPEG0007653166000242.jpg230160JPEG0007653166000243.jpg245160JPEG0007653166 000244.jpg245160JPEG0007653166000245.jpg246160JPEG0007653166000246.jpg246160JPEG0007653166000247.jpg72160

[0169] Example 12 JPEG0007653166000248.jpg41147JPEG0007653166000249.jpg54107

[0170] Reference for the preparation method of compound 12-1: Journal of Medicinal Chemistry (2007), 50(3), 566-584. LC-MS(ESI):m / z 354.1[M-18] + . 1 H NMR (400MHz, DMSO-d6) δ=8.95(d,J=8.4Hz,1H),8.11-7.90(m,5H),7.74(d,J=8.0Hz,1H),7.65-7.56(m,2H),7.54-7.48(m,1H).

[0171] Compound 12-1 (1 g, 2.69 mmol) is added to a solution of compound 12-3 (915 mg, 5.38 mmol) in pyridine (14 mL). The mixture is stirred at 30° C. for 16 h and concentrated. The residue is purified by flash column chromatography (petroleum ether / ethyl acetate=1 / 1, 30% tetrahydrofuran in petroleum ether) to give compound 12-2 (1.36 g, 2.69 mmol) as a yellow solid. LC-MS(ESI): m / z 506.2[M+H] + ,

[0172] A solution of compound 12-2 (420 mg, 0.67 mmol) and (0.38 mL, 6.65 mmol) in methanol (3 mL) is stirred at 70° C. for 2 h. The mixture is filtered and the solid is washed with methanol (20 mL). The filtrate is concentrated to give the crude product. The residue is purified by preparative HPLC (column: Agela Dura Shell NH2 150 mm×30 mm×5 um, mobile phase: heptane-ethanol, B%: 50%-100%, 10 min) to give yellow oily compound 12-3 (200 mg, 0.53 mmol, 80.2%). LC-MS(ESI):m / z 376.2[M+H] + .

[0173] To a solution of compound 12-3 (80 mg, 0.21 mmol), 4-chloroquinazoline (5.3 mg, 0.032 mmol) in i-PrOH (0.2 mL) is added CSA (76 μL, 0.43 mmol). The mixture is heated to 80 °C and stirred for 16 h. To the mixture is added HO (10 mL) and DCM (10 mL). The aqueous layer is separated and extracted with DCM (10 mL x 2). The combined organic layer is concentrated and purified by preparative HPLC (column: Phenomenex Gemini-NX 150 x 30 mm x 5 um, mobile phase: water (0.05% ammonia water v / v)-acetonitrile, B%: 30%-70%, 10 min) to give compound 114 (5.12 mg, 0.01 mmol, 4.6%) as a yellow solid.

[0174] A solution of compound 12-3 (50 mg, 0.13 mmol), 4-chloroquinazoline (48 mg, 0.29 mmol) in i-PrOH (1.6 mL) is heated to 100° C. and stirred for 16 h. The mixture is concentrated to obtain the crude product. The residue is purified by preparative HPLC (column: Phenomenex Gemini C18 250×50 mm×10 um, mobile phase: [water (0.05% ammonia water v / v)-acetonitrile], B%: 35%-55%, 10 min) to obtain yellow solid compound 115 (40 mg, 0.08 mmol, 59.3%).

[0175] The following compounds are synthesized according to the steps described in Example 12. JPEG0007653166000250.jpg236160JPEG0007653166000251.jpg246160JPEG0007653166000252.jpg218160 JPEG0007653166000253.jpg246160JPEG0007653166000254.jpg246160JPEG0007653166000255.jpg155160

[0176] Example 13 JPEG0007653166000256.jpg42143

[0177] Compound 13-1 is prepared according to the method for preparing C1 using 4-bromo-1-naphthylamine as the raw material to give a white solid product. n-BuLi (2.5M in tetrahydrofuran, 0.72mL, 1.80mmol) is cooled to -70°C and a suspension of compound 13-1 (279mg, 0.82mmol) in tetrahydrofuran (2mL) is added under nitrogen gas. The mixture is stirred at -70°C for 2 hours. A solution of α,α-diphenyl-N-sulfinyl-benzylmethylamine (301mg, 0.98mmol) in tetrahydrofuran (2mL) is added to the mixture and stirred at -70°C for 25 minutes, then at 0°C for 10 minutes. t-BuOCl (98mg, 0.90mmol) is added and the mixture is stirred at 0°C for 15 minutes. Triethylamine (114μL, 0.82mmol) and compound 13-2 (125mg, 0.98mmol) are then added dropwise. The reaction mixture is stirred at 25°C for 16 hours. MsOH (0.27 mL, 4.100 mmol) is added and the mixture is stirred at 25° C. for 0.5 h. The reaction is quenched with saturated NaHCO3 (20 mL) solution and the mixture is extracted with DCM (20 mL×3). The combined organic layers are dried over MgSO4, and the filtrate is filtered and concentrated to give a yellow semi-solid crude product. The crude product is washed with ethyl acetate (5 mL+2 mL) to give a white solid (90 mg). The solid is purified by preparative HPLC (column: Agela DuraShell NH2 150 mm×30 mm×5 um, mobile phase: heptane-ethanol, B%: 5-95%, 10 min) to give a white solid (6 mg). The solid is washed with ethanol (0.5 mL) and dried under vacuum to give pure compound 123 (1.8 mg, 0.4%) as a white solid.

[0178] JPEG0007653166000257.jpg145160

[0179] Example 14 JPEG0007653166000258.jpg45140

[0180] To a solution of compound 14-1 (300 mg, 0.94 mmol) in pyridine (3 mL) at 0° C., C1 (338 mg, 0.94 mmol) is added, and the mixture is stirred at 25° C. for 12 h, turning brown. The mixture is concentrated. Water (20 mL) is added, and extracted with DCM (20 mL×3). The combined organic phase is dried over Na2SO4, the filtrate is filtered, and concentrated to give crude compound 14-2 (450 mg, 0.92 mmol, 97.9%) as a brown oil. LC-MS(ESI): m / z 490.1[M+H] + .

[0181] To a solution of compound 14-2 (450 mg, 0.92 mmol) in methanol (20 mL), add 10% Pd / C (100 mg, 0.61 mmol), and stir the mixture at 15 Psi H2 at 25 °C for 12 h. Filter the mixture and wash with DCM (100 mL). Concentrate the filtrate. Purify the residue by preparative HPLC (column: YMC-Actus Triart C18 150 x 30 mm x 5 um, mobile phase: water (0.05% ammonia water v / v)-acetonitrile, B%: 48%-68%, 10 min) to obtain white solid compound 124 (50 mg, 0.1 mmol).

[0182] Acetyl chloride (5 μL, 0.064 mmol) is added to a solution of compound 124 (50 mg, 0.053 mmol) in pyridine (3 mL) at 0° C. The mixture is stirred for 12 h at 25° C. and turns brown. LCMS shows the reaction is complete. The mixture is concentrated. The residue is purified by preparative HPLC (column: YMC Triart C18 150×25 mm×5 um, mobile phase: water (0.05% ammonia v / v)-acetonitrile, B%: 42%-62%, 10 min) to give compound 125 (2 mg, 0.004 mmol, 7.5%) as a white solid.

[0183] JPEG0007653166000259.jpg236160JPEG0007653166000260.jpg101151

[0184] Example 15 JPEG0007653166000261.jpg57141

[0185] To a solution of compound 15-1 (50 mg, 0.21 mmol) in methanol (0.8 mL) and H2O (0.2 mL) is added chlorohydrogen, hydroxylamine (21.6 mg, 0.31 mmol) and NaOAc (25.5 mg, 0.31 mmol). The suspension is stirred at 20 °C for 16 h. The mixture is added with H2O (5 mL) and extracted with DCM (5 mL x 3). The combined organic layer is dried over Na2SO4, the filtrate is filtered and concentrated to give compound 15-2 (60 mg, 0.21 mmol, 99.4%) as a colorless oil. LC-MS(ESI): m / z 201.2[M-56] + .

[0186] To a solution of compound 15-2 (60 mg, 0.21 mmol) and NHOH (0.1 mL) in methanol (20 mL), RANEY NICKEL (0.02 mL, 0.275 mmol) is added and the mixture is stirred for 16 h under 15 Psi of H2 at 25° C. The mixture is filtered and the filtrate is concentrated to give the desired product compound 15-3 (42 mg, 0.16 mmol, 60.0%) as a colorless oil. LC-MS(ESI):m / z 187.2[M-56] + .

[0187] To a solution of DABCO (58.2 mg, 0.52 mmol), compound 15-3 (42 mg, 0.17 mmol) in DCM (1.5 mL), C1 (81 mg, 0.23 mmol) is added, and the mixture is stirred at 20° C. for 3 h. The mixture is diluted with water (3 mL) and extracted with DCM (3 mL×3). The organic matter is dried over MgSO4, filtered, and the filtrate is concentrated under vacuum to give compound 127 (60 mg, 0.10 mmol, 55.1%) as a yellow oil. The crude product (20 mg) is purified by preparative high performance liquid chromatography (column: Phenomenex Gemini C18 250×50 mm×10 um, mobile phase: water (0.05% aqueous ammonia v / v)-acetonitrile, B%: 50%-70%, 10 min) to give compound 127 (7.5 mg, 2.6%) as a white solid.

[0188] To a solution of compound 127 (60 mg, 0.11 mmol) in DCM (0.5 mL), add HCl / dioxane (0.5 mL, 4N, 2 mmol) and stir the mixture for 16 h at 20° C. The mixture is concentrated and purified by preparative HPLC (column: Phenomenex Gemini-NX 150×30 mm×5 um, mobile phase: water (0.225% formic acid)-acetonitrile, B%: 17%-37%, 10 min) to obtain white solid 2-methyl-N-(4-{[1-(piperidin-4-yl)propan-2-yl]sulfamoyl}naphthalen-1-yl)benzamide (24 mg, 0.05 mmol, 48.4%).

[0189] The following compounds are synthesized according to the steps described in Example 15. JPEG0007653166000262.jpg210160JPEG0007653166000263.jpg246160JPEG000 7653166000264.jpg246160JPEG0007653166000265.jpg233160JPEG00076531660 00266.jpg246160JPEG0007653166000267.jpg246160JPEG0007653166000268.j pg246160JPEG0007653166000269.jpg246160JPEG0007653166000270.jpg203160

[0190] Example 16 TIFF0007653166000271.tif64145TIFF0007653166000272.tif35144

[0191] To a solution of compound 13-2 (5.6 g, 23.43 mmol) and triethylamine (9.7 mL, 70.29 mmol) in DCM (100 mL) is added compound 16-1 (0.2 mL, 1.45 mmol). The mixture is stirred at 30° C. for 16 h. Water (50 mL) is added to the mixture, and the organic layer is separated and concentrated. The residue is purified by flash silica gel chromatography (ISCO, 40 g SepaFlash Silica Flash Column, eluent: 0-22% ethyl acetate / petroleum ether gradient @ 40 mL / min) to give compound 16-2 (1 g, 22.1%) as a yellow oil. LC-MS(ESI):m / z 344.9[MH] + ,

[0192] A solution of compound 16-2 (100 mg, 0.29 mmol), aqueous ammonia (28% aqueous solution) (1 mL, 7.27 mmol) in n-BuOH (1 mL) was stirred at 100° C. for 16 h. The mixture was concentrated to give compound 16-3 (104 mg, 0.29 mmol, 99%) as a yellow solid. LC-MS(ESI):m / z 326.0[MH] + ,

[0193] A solution of compound 16-3 (0.35 g, 1.07 mmol), Pd / C (0.3 g, 1.07 mmol), NH3.H2O (1 mL) in methanol (40 mL) is stirred under H2 at 30° C. for 16 h. The mixture is filtered and the filtrate is concentrated to give the desired product compound 16-3 (300 mg, 0.91 mmol, 84.9%) as a yellow oil. LC-MS(ESI):m / z 298.1[MH] + ,

[0194] To a mixture of compound 16-4 (60 mg, 0.20 mmol) and 2-phenylacetic acid (0.04 mL, 0.30 mmol) in dioxane (2 mL), add T3P (0.36 mL, 0.61 mmol) and DIEA (0.20 mL, 1.21 mmol). Stir the mixture at 100 °C for 16 h. Concentrate the mixture to obtain the crude product. Purify the residue by preparative HPLC (column: Phenomenex Gemini C18 250 × 50 mm × 10 um, mobile phase: water (0.05% aqueous ammonia v / v)-acetonitrile, B%: 48%-68%, 10 min) to obtain the desired product as a white solid compound 144 (2.2 mg, 2.4%).

[0195] Compound 16-4 (67 mg, 0.23 mmol) and isothiocyanatobenzene (32 μL, 0.27 mmol) in pyridine (1 mL) were stirred for 16 h at 30° C. The mixture was concentrated to give the crude product compound 16-5 (80 mg, 0.11 mmol, 49.3%) as a yellow oil. LC-MS(ESI): m / z 433.3[M+H] + ,

[0196] To a solution of compound 16-5 (80 mg, 0.11 mmol) in ethanol (0.3 mL), EDC HCl (32 mg, 0.17 mmol) is added and the mixture is stirred at 70 °C for 2 h. The mixture is concentrated to obtain the crude product. The residue is purified by preparative HPLC (column: Phenomenex Gemini C18 250 × 50 mm × 10 um, mobile phase: water (0.05% ammonia water v / v)-acetonitrile, B%: 50%-70%, 10 min) to obtain compound 145 (10.11 mg, 0.02 mmol, 22.2%) as a white solid.

[0197] The following compounds are synthesized according to the steps described in Example 16. JPEG0007653166000273.jpg233160JPEG0007653166000274.jpg167160

[0198] Example 17 JPEG0007653166000275.jpg86142

[0199] At 0°C, DIEA (0.4mL, 2.36mmol) is added in one portion to a solution of compound 13-2 (100mg, 0.79mmol) in tetrahydrofuran (2mL), and then added dropwise to a solution of compound 17-1 (377mg, 0.87mmol) in tetrahydrofuran (2mL). The reaction mixture is heated to 25°C and stirred for 3 hours. 10mL of water is added to the reaction mixture to quench the reaction. The reaction mixture is extracted with DCM (5mL x 3). The combined organic layers are dried over Na2SO4, filtered, and the filtrate is concentrated under reduced pressure. The residue is purified by preparative thin layer chromatography (petroleum ether:ethyl acetate = 2:1) to give compound 17-2 (170mg, 0.43mmol, 54.6%) as a yellow solid. LC-MS: (ESI+) m / z 398.1 [M+H] + ,

[0200] To a solution of compound 17-2 (50 mg, 0.13 mmol), ethynyltrimethylsilane (20 μL, 0.15 mmol), CuI (1.2 mg, 6 μmol), triphenylphosphine (3.3 mg, 0.013 mmol) and triethylamine (0.15 mL, 1.08 mmol) in pyridine (1 mL) at 25° C., PdCl2(PPh3)2 (3.99 mg, 0.006 mmol) is added in one portion. The reaction mixture is heated to 80° C. and stirred for 16 h. The reaction mixture is cooled to room temperature and 5 mL of water is added, and extracted with DCM (1 mL×4). The combined organic layer is dried over Na2SO4 and filtered. The filtrate is concentrated under reduced pressure to give a residue. The residue was purified by preparative thin-layer chromatography (petroleum ether:ethyl acetate (V / V)=2:1) ​​to give compound 17-3 (70 mg, 0.12 mmol, 93.9%) as a yellow solid. LC-MS: (ESI+) m / z 414.3 [M+H] + ,

[0201] To a solution of compound 17-3 (70 mg, 0.17 mmol) in a mixture of methanol (2 mL) and water (1 mL), K2CO3 (25.7 mg, 0.19 mmol) is added in one portion. The reaction mixture is then stirred at 25 °C for 16 h. 5 mL of water is added to the reaction mixture and extracted with DCM (1 mL x 4). The combined organic layer is dried over Na2SO4 and filtered. The filtrate is concentrated under reduced pressure to give a residue of pale green oil compound 17-4 (50 mg, 0.15 mmol, 83.3%). The crude product is used in the next step without further purification. LC-MS: (ESI+) m / z 342.1 [M+H] + ,

[0202] To a solution of compound 17-4 (30 mg, 0.088 mmol) in N,N-dimethylformamide (2 mL), sodium azide (22.8 mg, 0.35 mmol) is added in one portion, the reaction mixture is heated to 100 °C, and stirred for 16 h. 5 mL of water is added to the reaction mixture, and the reaction mixture is adjusted to PH=9. The aqueous layer is poured into sodium hypochlorite solution, and extracted with ethyl acetate (5 mL x 3). The combined organic layers are dried over Na2SO4 and filtered. The filtrate is concentrated under reduced pressure. The residue is purified by preparative HPLC (column: Phenomenex Gemini C18 250 x 50 mm x 10 um [water (0.05% ammonia water v / v)-acetonitrile] B%: 40%-60%, 10 min) to give compound 147 (1.11 mg, 0.00 mmol, 3.3%) as a white solid.

[0203] The following compounds are synthesized according to the steps described in Example 17. JPEG0007653166000276.jpg193160

[0204] Example 18 JPEG0007653166000277.jpg54145

[0205] Under nitrogen gas, to a solution of compound 17-2 (50 mg, 0.13 mmol) and compound 18-1 (65.9 mg, 0.25 mmol) in a mixed solvent of dioxane (3 mL) and water (1 mL), K3PO4 (80.3 mg, 0.38 mmol) and Pd(dppf)Cl2 (18.5 mg, 0.025 mmol) are added in one portion. The reaction mixture is then heated to 80°C and stirred for 16 hours. The reaction mixture is cooled to 25°C and filtered. The filtrate is concentrated under reduced pressure to give a residue of brown oily compound 18-2 (100 mg, 0.11 mmol, 89.3%), which is used in the next step without further purification. LC-MS: (ESI+) m / z 533.3 [M+H] + ,

[0206] Compound 18-2 (50 mg, 0.094 mmol) is added in one portion in HCl / dioxane (4N, 1 mL) at 25° C. The reaction mixture is stirred for 48 h at 25° C. The reaction mixture is concentrated under reduced pressure to give a residue, which is purified by preparative HPLC (column: YMC Triart C18 150×25 mm×5 um [water (0.225% formic acid)-acetonitrile] B%: 73%-93%, 10 min) to give compound 149 (4.70 mg, 0.01 mmol, 11.6%) as a white solid.

[0207] JPEG0007653166000278.jpg131160

[0208] Example 19 JPEG0007653166000279.jpg38142

[0209] Compound 19-1 (200 mg, 0.82 mmol) is added to a solution of compound 13-2 (124.7 mg, 0.98 mmol) in pyridine (2 mL) at 0° C. The mixture is stirred at 20° C. for 12 h. The mixture is concentrated. The residue is purified by preparative HPLC (column: YMC-Actus Triart C18 150×30 mm×5 um, mobile phase: water (0.05% ammonia water v / v)-acetonitrile, B%: 65%-85%, 10 min) to give yellow oily compound 19-2 (90 mg, 0.27 mmol, 33.3%). LC-MS(ESI):m / z 336.1[M+H] + ,

[0210] To a solution of compound 19-2 (140 mg, 0.42 mmol) in DMSO (14 mL), add NaCN (102.3 mg, 2.09 mmol). Stir the mixture at 100° C. for 3 h. Add water (50 mL) and extract with DCM (50 mL×3). Dry the combined organic phase over Na2SO4, filter and concentrate the filtrate to give crude compound 19-3 (150 mg, 0.44 mmol) as a brown solid. LC-MS(ESI):m / z 365.1[M+Na] + ,

[0211] To a solution of compound 19-3 (150 mg, 0.44 mmol) and dibutylstannanone (15 μL, 0.088 mmol) in toluene (8 mL), add trimethylsilyl azide (115 μL, 0.88 mmol) and stir the mixture at 120 °C for 15 h. Concentrate the mixture. Purify the residue by preparative HPLC (column: Phenomenex Gemini NX-C18 (75 × 30 mm × 3 um), mobile phase: water (0.05% aqueous ammonia v / v)-acetonitrile, B%: 10% to 50%, 10 min) to obtain white solid compound 150 (5 mg, 0.01 mmol, 2.9%).

[0212] JPEG0007653166000280.jpg153160

[0213] Example 20 JPEG0007653166000281.jpg83144

[0214] To a solution of compound 13-2 (100 mg, 0.79 mmol) in pyridine (5 mL, 61.9 mmol) is added compound 20-1 (191 mg, 0.79 mmol) in one portion. The reaction mixture is stirred at 25 °C for 16 h. The reaction mixture is quenched by adding 15 mL of water. The mixture is extracted with DCM (5 mL x 3). The combined organic layers are dried over Na2SO4, then filtered, and the filtrate is concentrated under reduced pressure to give a residue of compound 20-1 (150 mg, 0.45 mmol, 57.2%) as a brown oil. The residue is used in the next step without further purification. LC-MS: (ESI+) m / z 334.2 [M+H] + ,

[0215] To a solution of compound 20-2 (100 mg, 0.18 mmol) and 1-(bromomethyl)-2-nitrobenzene (40.8 mg, 0.19 mmol) in acetonitrile (2 mL), K2CO3 (124 mg, 0.90 mmol) and NaI (5.40 mg, 0.036 mmol) are added in one portion, then the reaction mixture is heated to 70° C. and stirred for 16 h. The reaction mixture is concentrated under reduced pressure to remove the solvent. Then 10 mL of water is added to the reaction mixture. The reaction mixture is extracted with DCM (3 mL×3). The combined organic layers are dried over Na2SO4 and filtered. The filtrate is concentrated under reduced pressure to obtain a residue, which is purified by preparative HPLC (column: YMC Triart C18 150×25 mm×5 um [water (0.225% FA)-acetonitrile] B%: 70%-90%, 10 min) to obtain white solid compound 20-3 (25 mg, 0.05 mmol, 29.7%). LC-MS: (ESI+) m / z 469.2 [M+H] + ,

[0216] To a solution of compound 20-3 (10 mg, 0.021 mmol) in AcOH (2 mL) at 25° C., zinc (20.6 mg, 0.32 mmol) is added in one portion, and the reaction mixture is stirred at 25° C. for 16 h. The reaction mixture is filtered, and the solid is washed with AcOH (1 mL). 10 mL of water is added to the filtrate, and the mixture is adjusted to PH=8 with saturated NaHCO3 solution. The mixture is extracted with DCM (3 mL×3). The combined organic layers are dried over MgSO4 and filtered. The filtrate is concentrated under reduced pressure to give a residue of yellow oil compound 20-4 (8 mg, 0.02 mmol, 85.5%). The residue is used in the next step without further purification. LC-MS: (ESI+) m / z 439.3 [M+H] +

[0217] Acetyl chloride (3.8 mg, 0.048 mmol) is added in one portion to a solution of compound 20-4 (8 mg, 0.032 mmol) in pyridine (1 mL) at 0° C. Then the reaction mixture is warmed to 25° C. and stirred for 1 h. A drop of water is added to the reaction mixture to quench the reaction. The reaction mixture is then concentrated under reduced pressure to obtain a residue, which is purified by preparative HPLC (column: YMC Triart C18 150×25 mm×5 um [water (0.225% formic acid)-acetonitrile] B%: 58%-78%, 10 min) to obtain white solid compound 151 (1.91 mg, 12.5%).

[0218] JPEG0007653166000282.jpg167160

[0219] Example 21 JPEG0007653166000283.jpg43131

[0220] Compound 21-1 (452 ​​mg, 1.77 mmol) is added to a solution of compound 13-2 (150 mg, 1.18 mmol) and DABCO (397 mg, 3.54 mmol) in DCM (3 mL) at 0° C. The mixture is stirred for 16 h at 20° C. The mixture is concentrated and suspended in 1M HCl (5 mL). The mixture is extracted with ethyl acetate (3 mL×3). The combined organic layers are dried over MgSO4, filtered and concentrated to give crude compound 21-2 (427 mg, 1.23 mmol) as a yellow solid. LC-MS(ESI):m / z 368.1,370.0[M+Na] + ,

[0221] Under nitrogen gas, to a solution of compound 21-2 (50 mg, 0.14 mmol) and proline (66 mg, 0.58 mmol) in DMSO (0.8 mL), add cesium carbonate (141 mg, 0.43 mmol) and CuI (5.5 mg, 0.029 mmol). Stir the mixture at 100°C for 16 h. Cool the mixture to room temperature and filter. Purify the filtrate by preparative HPLC (column: YMC Triart C18 150 x 25 mm x 5 um, mobile phase: water (0.05% ammonia water v / v)-acetonitrile, B%: 20%-40%, 10 min) to obtain white solid compound 152 (5.94 mg, 0.02 mmol, 10.8%).

[0222] The following compounds are synthesized according to the steps described in Example 21. JPEG0007653166000284.jpg236160JPEG0007653166000285.jpg163160

[0223] Example 22 JPEG0007653166000286.jpg47145

[0224] To a solution of compound 17 (190 mg, 0.42 mmol), triethylamine (175 μL, 1.26 mmol) and (t-butoxycarbonyl)glycine (96 mg, 0.55 mmol) in acetonitrile (10 mL) at 0° C., add T3P (401.6 mg, 0.63 mmol). Stir the mixture at 30° C. for 4 h. Concentrate the mixture. Add water (20 mL) and extract with DCM (20 mL×3). Dry the combined organic phase over Na2SO4, filter and concentrate. The crude product is purified by supercritical fluid chromatography (column: DAICEL CHIRALPAK AS (250 mm × 30 mm, 10 um), mobile phase: 0.1% NH3H2O ​​EtOH, B%: 40% to 40%) to obtain white solid P1 (compound 155A, 60 mg), white solid P2 (compound 155B, 30 mg), white solid P3 (compound 155C, 50 mg), white solid P4 (compound 155D, 30 mg).

[0225] To a solution of compound 155A (50 mg, 0.082 mmol) in DCM (2 mL) is added HCl / dioxane (4N, 0.5 mL) at 0° C. The mixture is stirred for 3 h at 30° C. The mixture is concentrated to give compound 156A (40 mg, 0.07 mmol, 89.5%) as a white solid.

[0226] The following compounds are synthesized according to the steps described in Example 22. JPEG0007653166000287.jpg236160JPEG0007653166000288.jpg243160JPEG0007653166000289.jpg213160JPEG000 7653166000290.jpg245160JPEG0007653166000291.jpg206160JPEG0007653166000292.jpg245160JPEG0007653166 000293.jpg241160JPEG0007653166000294.jpg220160JPEG0007653166000295.jpg245160JPEG0007653166000296. jpg241160JPEG0007653166000297.jpg211160JPEG0007653166000298.jpg246160JPEG0007653166000299.jpg51160

[0227] Example 23 JPEG0007653166000300.jpg87113

[0228] Compound 23-4 (0.84 mL, 6.41 mmol) is added to a solution of compound 23-1 (1 g, 5.34 mmol) in pyridine (10 mL, 123 mmol) at 25° C., then the reaction mixture is heated to 100° C. and stirred for 16 h. The reaction mixture is concentrated under reduced pressure to give a residue. The residue is recrystallized with DCM (15 mL) and petroleum ether (2 mL) to give compound 23-2 (1.5 g, 3.90 mmol, 73.2%) as a white solid. LC-MS: (ESI+) m / z 306.1 [M+H] + ,

[0229] Compound 23-5 (0.79 g, 4.29 mmol) is added in 10 portions to a suspension of compound 23-2 (1.5 g, 3.90 mmol) in acetone (30 mL) at 25° C., followed by dropwise addition of triethylamine (0.6 mL, 4.29 mmol) within 5 min. The reaction mixture is heated to 80° C. and stirred at 80° C. for 16 h. The reaction mixture is filtered, and the filtrate is concentrated under reduced pressure to give a residue of compound 23-3 (0.7 g, 2.20 mmol 56%). The residue is used in the next step without further purification. LC-MS: (ESI+) m / z 375.2 [M+H] + , (morpholinequenched)

[0230] Compound 23-3 (345.6 mg, 0.85 mmol) is added in one portion to a solution of compound 23-6 (150 mg, 0.66 mmol) and DABCO (294.3 mg, 2.63 mmol) in DCM (3 mL) at 25° C. Then the reaction mixture is stirred for 16 h at 25° C. 3 mL of water is added to the reaction mixture and extracted with DCM (1 mL×3). The combined organic layer is dried over Na2SO4 and then filtered. The filtrate is concentrated under reduced pressure to give a residue. The residue is purified by preparative HPLC (column: Welch Xtimate C18 100×25 mm×3 um [water (0.05% aqueous ammonia v / v)-acetonitrile] B%: 29%-69%, 9 min) to give compound 160 (300 mg, 0.58 mmol, 88.6%) as a white solid.

[0231] To a solution of compound 160 (150 mg, 0.66 mmol) in DCM (3 mL) is added HCl / dioxane (4N, 0.8 mL) in one portion at 25° C. The reaction mixture is then stirred for 3 h at 25° C. The mixture is concentrated to give compound 161A (153 mg, 0.31 mmol, 99.7%) as a white solid.

[0232] The following compounds are synthesized according to the steps described in Example 23. JPEG0007653166000301.jpg236160JPEG0007653166000302.jpg230160JPEG0007653166000303.jpg246160JPEG00076531660 00304.jpg246160JPEG0007653166000305.jpg246160JPEG0007653166000306.jpg246160JPEG0007653166000307.jpg155160

[0233] Example 24 JPEG0007653166000308.jpg60145

[0234] To a solution of compound 24-1 (5 g, 0.035 mol) and triethylamine (14.7 mL, 0.106 mol) in DCM (50 mL) under 0° C., acetyl chloride (3.8 mL, 0.053 mol) is added dropwise over 10 min. The reaction mixture is then warmed to 25° C. and stirred for 16 h. The reaction mixture is quenched with 1 mL of water. 50 mL of saturated NH4Cl solution is then added to the reaction mixture and stirred for 10 min. The reaction mixture is extracted with DCM (20 mL×3). The combined organic layers are dried over Na2SO4 and filtered. The filtrate is concentrated under reduced pressure. The residue is recrystallized with DCM (15 mL) and petroleum ether (5 mL) to give compound 24-1 (4 g, 21.78 mmol, 61.7%) as a white solid. The solid is used in the next step without further purification. LC-MS: (ESI+) m / z 184.1 [M+H] +

[0235] Compound 24-2 (3 g, 0.016 mol) is added to ClSO3H (5.44 mL, 0.082 mol) in batches within 30 min under 0°C. Then NaCl (0.86 g, 0.015 mol) is added to the reaction mixture in 10 portions under 0°C. The reaction mixture is then heated to 90°C and stirred for 1 h. The reaction mixture is cooled to 25°C and slowly added to ice to quench the reaction, keeping the temperature below 0°C. The precipitated solid is filtered and dried under vacuum to obtain compound 24-3 (3.1 g, 10.99 mmol, 67.2%) as a white solid. LC-MS: (ESI+) m / z 346.1 [M+H] + (1-methylpiperazine quenched)

[0236] Compound 24-3 (345 mg, 0.85 mmol) is added in one portion to a solution of compound 23-6 (100 mg, 0.44 mmol) and DABCO (195 μL, 1.75 mmol) in DCM (3 mL) at 25° C. The reaction mixture is then stirred for 3 h at 25° C. 3 mL of water is added to the mixture and extracted with DCM (1 mL×3). The combined organic layers are dried over Na2SO4 and then filtered. The filtrate is concentrated under reduced pressure to give compound 24-4 (150 mg, 0.32 mmol, 72.3%) as a yellow solid. The crude product is used in the next step without further purification. LC-MS:(ESI+)m / z 374.1[M-100+H] + .

[0237] To a solution of compound 24-4 (150 mg, 0.32 mmol) in ethanol (2 mL) at 25° C., NaOH (75.9 mg, 1.90 mmol) is added in one portion. The reaction mixture is then heated to 100° C. and stirred for 3 h. The reaction mixture is concentrated under reduced pressure. 3 mL of water is added to the residue, and the mixture is extracted with ethyl acetate (1 mL×3). The combined organic layers are dried over Na2SO4 and filtered. The filtrate is concentrated under reduced pressure to give a residue of compound 24-5 (130 mg) as a yellow solid. The crude product is used in the next step without further purification. LC-MS:(ESI+)m / z 332.1[M-100+H]+ ,

[0238] Compound 23-4 (143 mg, 0.93 mmol) is added to a solution of compound 24-5 (100 mg, 0.23 mmol), triethylamine (0.19 mL, 1.38 mmol) and DMAP (5.7 mg, 0.046 mmol) in DMAP (2 mL) at 0° C. The mixture is heated to 80° C. and stirred for 18 hours. The reaction mixture is then cooled to 25° C., and sodium hydroxide solution (2 M aqueous solution, 0.9 mL, 1.8 mmol) is added to the reaction mixture. The mixture is stirred for 16 hours at 25° C. and extracted with ethyl acetate (5 mL×3). The combined organic layer is dried over Na2SO4 and filtered. The filtrate is concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (column: Agela DuraShell C18 150×25 mm×5 um [water (0.225% formic acid)-acetonitrile] B%: 37%-72%, 10 min) to give compound 168 (50 mg, 0.09 mmol), 39.3%) as a white solid.

[0239] To a solution of compound 168 (50 mg, 0.091 mmol) in DCM (5 mL) is added HCl / dioxane (4 M, 2 mL) at 25° C., and the reaction mixture is stirred for 3 h at 25° C. The reaction mixture is concentrated to give compound 169A (21.29 mg, 0.04 mmol, 48.1%) as a white solid.

[0240] The following compounds are synthesized according to the steps described in Example 24. JPEG0007653166000309.jpg236160JPEG0007653166000310.jpg246160JPEG00076531660 00311.jpg246160JPEG0007653166000312.jpg246160JPEG0007653166000313.jpg233160 JPEG0007653166000314.jpg246160JPEG0007653166000315.jpg246160JPEG00076531660 00316.jpg246160JPEG0007653166000317.jpg233160JPEG0007653166000318.jpg119160

[0241] Example 25 JPEG0007653166000319.jpg70146

[0242] Compound 25-1 (5.0 g, 24.3 mmol) is added to a solution of concentrated HCl (20 mL, 235 mmol) diluted with water (0.5 mL). The mixture is cooled to 0° C., and a solution of sodium nitrite (1.84 g, 26.7 mmol) in water (1 mL) is added dropwise within 2 min, while maintaining the temperature at 0° C. After the addition, the slurry is stirred at 0° C. for 40 min to obtain a brown suspension. CuSO4.5H2O (6.06 g, 24.3 mmol) is added to concentrated HCl (20 mL, 235 mmol) and cooled to 0° C. NaHSO3 (5.05 g, 48.5 mmol) is dissolved in water (2 mL), and the solution is divided into two portions, and the first portion is added to the hydrochloric acid solution. The brown suspension is added dropwise to the hydrochloric acid solution, while maintaining the temperature below 5° C. At the same time, another portion of NaHSO3 solution is added dropwise. The reaction mixture is then stirred at 0°C for 1 h until no more gas is formed. The reaction mixture is filtered to obtain a filter cake, and the filtrate is extracted with DCM (20 mL x 3). The combined organic layers are dried over Na2SO4 and then filtered. The filtrate is concentrated under reduced pressure to obtain a residue. The residue is purified by flash column chromatography (petroleum ether:ethyl acetate, 1:0 to 1:2) to obtain compound 25-2 (1 g, 2.76 mmol, 11.4%) as a yellow solid. LC-MS: (ESI+) m / z 354.1 [M+H] + (1-methylpiperazinequenched),

[0243] Compound 25-2 (345 mg, 0.85 mmol) is added in one portion to a solution of compound 23-6 (100 mg, 0.44 mmol) and DABCO (196 mg, 1.75 mmol) in DCM (3 mL) at 25° C. Then the reaction mixture is stirred for 3 h at 25° C. 3 mL of water is added to the mixture and extracted with DCM (1 mL×3). The combined organic layers are dried over Na2SO4 and filtered. The filtrate is concentrated under reduced pressure to give a residue of compound 25-3 (160 mg, 0.33 mmol, 75.9%) as a yellow solid. The residue is used in the next step without further purification. LC-MS: (ESI+) m / z 426.1 [M+H-56] + ,

[0244] To a solution of compound 25-3 (100 mg, 0.21 mmol) in methanol (30 mL) is added Pd / C (50 mg) in one portion. The reaction mixture is then stirred under H2 (15 Psi) at 25 °C for 16 h. The reaction mixture is filtered and the filtrate is concentrated under reduced pressure to give compound 25-4 (88 mg) as a brown oil. LC-MS:(ESI+)m / z 352.1[M+H-100] + ,

[0245] Compound 23-4 (103 mg, 0.66 mmol) is added in one portion to a solution of compound 25-4 (100 mg, 0.22 mmol) in pyridine (1 mL, 12.4 mmol) at 25°C. The reaction mixture is then heated to 100°C and stirred at 100°C for 18 hours. The reaction mixture is then cooled to 25°C and 1 mL of water is added to quench the reaction. The reaction mixture is then concentrated under reduced pressure to obtain a residue. The residue is purified by preparative HPLC (Phenomenex Gemini C18 250 x 50 mm x 10 um, mobile phase: water (0.05% ammonia water v / v)-acetonitrile, B%: 55%-75%, 10 min) to obtain white solid compound 185 (17 mg, 0.03 mmol, 13.5%).

[0246] A solution of compound 185 (11 mg, 0.019 mmol) in DCM (2 mL, 31.1 mmol) is added in one portion to HCl / dioxane (4 M, 1 mL), and the reaction mixture is stirred at 25° C. for 18 h. The reaction mixture is concentrated under reduced pressure to give a residue. The residue is then added with 3 mL of water to give a colorless solution. The solution is lyophilized to give compound 186A (9.71 mg, 0.02 mmol, 99.4%) as a white solid.

[0247] The following compounds are synthesized according to the steps described in Example 25. JPEG0007653166000320.jpg214160JPEG0007653166000321.jpg245160JPEG000 7653166000322.jpg246160JPEG0007653166000323.jpg246160JPEG00076531660 00324.jpg246160JPEG0007653166000325.jpg246160JPEG0007653166000326.j pg246160JPEG0007653166000327.jpg246160JPEG0007653166000328.jpg227160

[0248] Example 26 JPEG0007653166000329.jpg46143JPEG0007653166000330.jpg4978

[0249] To a solution of compound 191 (40 mg, 0.069 mmol) and Pd(PPh3)4 (23.9 mg, 0.021 mmol) in N,N-dimethylformamide (2 mL), Zn(CN)2 (24.3 mg, 0.21 mmol) is added. The mixture is stirred at 110°C for 12 hours. The mixture is cooled to room temperature and filtered. The filtrate is concentrated. The residue is purified by preparative HPLC (column: YMC Triart C18 150 x 25 mm x 5 um, mobile phase: water (0.05% ammonia water v / v)-acetonitrile, B%: 53%-73%, 10 min) to obtain white solid compound 193 (20 mg, 0.04 mmol, 55.1%).

[0250] To a solution of compound 193 (15 mg, 0.028 mmol) in DCM (1 mL) at 0° C., add 4M HCl / dioxane (0.07 mL, 0.28 mmol). Stir the mixture at 30° C. for 2 h. Concentrate the mixture. Purify the residue by preparative HPLC (column: YMC Triart C18 150×25 mm×5 um, mobile phase: water (0.05% aqueous ammonia v / v)-acetonitrile, B%: 58%-78%, 10 min) to obtain compound 194 (4 mg, 0.01 mmol, 32.1%) as a white solid.

[0251] To a solution of compound 193 (100 mg, 0.19 mmol) in TFE (3 mL), add fluoroboric acid in ether (31 μL, 0.23 mmol). The mixture is stirred at 0° C. for 1 h, then at 20° C. for 2 h. The mixture is diluted with water (3 mL) and lyophilized. The crude product is purified by preparative HPLC (column: Phenomenex Gemini-NX C18 80×30 mm×5 um, mobile phase: water (0.05% ammonia v / v)-acetonitrile, B%: 43%-63%, 10 min) to obtain compound 195 (50 mg, 0.12 mmol, 61.7%) as a white solid.

[0252] The following compounds are synthesized according to the steps described in Example 26. JPEG0007653166000331.jpg236160JPEG0007653166000332.jpg246160JPEG0007653166000333.jpg22160

[0253] Example 27 JPEG0007653166000334.jpg42144

[0254] To a mixture of compound 191 (60 mg, 0.1 mmol) and 4,4,5,5-tetramethyl-2-(prop-1-en-2-yl)-1,3,2-dioxaborane (51 mg, 0.3 mmol) in dioxane (1 mL) is added a solution of Na2CO3 (33 mg, 0.6 mmol) in water (0.6 mL). Then Pd(dppf)Cl2CH2Cl2 (18 mg, 21 μmol) is added under N2. The mixture is stirred at 100 °C for 16 h. The mixture is diluted with water (5 mL) and DCM (2 mL). The mixture is filtered and the solid is washed with DCM (2 mL). The filtrate layers are separated and the aqueous layer is extracted with DCM (2 mL × 3). The combined organic layers are dried over Na2SO4, filtered and concentrated to give crude compound 27-1 (118 mg) as a black oil. LC-MS(ESI): m / z 442.3[M-100+H] + ,

[0255] To a mixture of compound 27-1 (118 mg, 0.1 mmol) and Pd(OAc)2 (0.32 mL, 1.96 mmol) in DCM (2 mL) under N2, add Pd(OAc)2 (132 mg, 0.6 mmol). Stir the mixture at 20°C for 16 h. Filter and concentrate the mixture. Purify the residue by preparative HPLC (column: Phenomenex Gemini-NX C18 75 x 30 mm x 3 um, mobile phase: water (0.225% formic acid)-acetonitrile, B%: 39%-79%, 10 min) to obtain white solid compound 27-2 (30 mg, 0.06 mmol, 56.3%). LC-MS(ESI): m / z 444.3[M-100+H] + ,

[0256] To a solution of compound 27-2 (30 mg, 0.055 mmol) in DCM (1 mL) at 0° C., 4M HCl / dioxane (0.14 mL, 0.56 mmol) is added and the mixture is stirred at 30° C. for 2 h. LCMS shows the reaction is complete. The mixture is concentrated to give compound 197A (25 mg, 0.05 mmol, 94.4%) as a white solid.

[0257] The following compounds are synthesized according to the steps described in Example 27. JPEG0007653166000335.jpg236160JPEG0007653166000336.jpg191160

[0258] Example 28 JPEG0007653166000337.jpg92144

[0259] Compound 28-1 (2 g, 14.03 mmol) is added to a solution of concentrated HCl (11.3 mL, 136.1 mmol) diluted with water (2 mL). The mixture is cooled to 0°C and a solution of NaNO2 (1.1 g, 15.4 mmol) in water (2 mL) is added dropwise within 2 min, while maintaining the temperature at 0°C. After the addition, the slurry is stirred at 0°C for 40 min to obtain a brown suspension. CuSO45H2O (3.9 g, 15.43 mmol) is added to HCl (11.3 mL, 136 mmol) and cooled to 0°C. NaHSO3 (2.92 g, 28.1 mmol) is dissolved in water (2 mL) and the solution is divided into two portions, where one portion is added to the hydrochloric acid solution and the other portion is reserved. The brown suspension is added dropwise to the HCl solution, while maintaining the temperature at 5°C. At the same time, another half of the NaHSO3 solution is added dropwise. The reaction mixture is then stirred at 0° C. for 1 h until no more gas is produced, and the reaction is complete. The reaction mixture is filtered to obtain a filter cake, and the filter cake is washed with DCM (300 mL). The combined organic layers are dried over Na2SO4 and then filtered. The filtrate is concentrated under reduced pressure to give compound 28-2 (2.3 g, 9.16 mmol, 65.3%) as a yellow oil.

[0260] To a solution of compound 23-6 (200 mg, 0.88 mmol) and compound 28-2 (297 mg, 1.3 mmol) in DCM (4 mL), add DABCO (147 mg, 1.3 mmol) and stir the mixture at 20 °C for 16 h. Concentrate the mixture. Purify the residue by flash silica gel chromatography (ISCO, 12 g SepaFlash Silica Flash Column, eluent: 0-50% ethyl acetate / petroleum ether gradient @ 20 mL / min) to give compound 28-3 (140 mg, 0.33 mmol), 38.2%) as a yellow oil. LC-MS(ESI): m / z 362.1[M-56+H] + ,

[0261] Compound 28-3 (20 mg, 0.05 mmol) and anti-N,N-dimethylcyclohexane-1,2-diamine (0.2 mg, 0.001 mmol) are added to a solution of compound 28-4 (10 mg, 0.08 mmol), CuI (0.5 mg, 0.002 mmol), potassium carbonate (13.3 mg, 0.1 mmol) in DGDE (0.5 mL) under nitrogen gas at 25 °C. The mixture is stirred at 160 °C for 2 h. The mixture is diluted with water (2 mL) and extracted with ethyl acetate (3 mL × 3). The combined organic phase is dried over MgSO4, filtered, and the filtrate is concentrated under vacuum to give a residue. The residue was purified by preparative HPLC (column: Phenomenex Gemini C18 250×50 mm×10 um, mobile phase: water (0.05% aqueous ammonia v / v)-acetonitrile, B%: 40%-60%, 10 min) to obtain white solid compound 28-5 (9 mg, 0.02 mmol, 34.6%). LC-MS(ESI): m / z 517.3[M+H] + ,

[0262] To a solution of compound 28-5 (9 mg, 0.017 mmol) in DCM (DCM) (0.5 mL) at 0° C., add HCl / dioxane (4 M, 0.1 mL) and stir the mixture at 25° C. for 2 h. The mixture is a white suspension. The mixture is concentrated under vacuum at 30° C. to give a residue. The residue is diluted with water (2 mL) and lyophilized to give compound 199A (4.76 mg, 0.01 mmol, 55.6%) as a yellow solid.

[0263] The following compounds are synthesized according to the steps described in Example 28. JPEG0007653166000338.jpg236160JPEG0007653166000339.jpg72160

[0264] Example 29 JPEG0007653166000340.jpg48103

[0265] Sodium cyanoborohydride (250 mg, 3.98 mmol) is added to a solution of compound 161A (100 mg, 0.17 mmol) and propan-2-one (74 μL, 1.01 mmol) in methanol (2 mL) at 25° C. The reaction mixture is heated to 50° C. and stirred at 50° C. for 16 hours. The reaction mixture is a white suspension. The reaction mixture is concentrated under reduced pressure to obtain a residue. The residue is purified by preparative HPLC (column: YMC Triart C18 150×25 mm×5 um [water (0.05% ammonia water v / v)-acetonitrile] B%: 50%-70%, 10 min) to obtain white solid compound 200 (13.70 mg, 0.03 mmol, 18.0%).

[0266] The following compounds are synthesized according to the steps described in Example 29. JPEG0007653166000341.jpg236160JPEG0007653166000342.jpg37160

[0267] Example 30 JPEG0007653166000343.jpg48103

[0268] To a solution of compound 161A (30 mg, 0.066 mmol) and DIEA (44 μL, 0.27 mmol) in N,N-dimethylformamide (0.6 mL), add 1-bromo-2-methoxyethane (13 μL, 0.14 mmol). Stir the mixture at 50 °C for 12 h. LCMS shows the reaction is complete. Concentrate the mixture. Purify the crude product by preparative HPLC (column: YMC Triart C18 150 × 25 mm × 5 um, mobile phase: water (0.05% ammonia v / v)-acetonitrile, B%: 44%-64%, 10 min) to obtain white solid compound 202 (22 mg, 0.05 mmol, 70.0%).

[0269] The following compounds are synthesized according to the steps described in Example 30. JPEG0007653166000344.jpg236160JPEG0007653166000345.jpg246160JPEG0007653166000346.jpg246160JPEG0007653166000347.jpg155160

[0270] Example 31 JPEG0007653166000348.jpg68142

[0271] Compound 31-1 (2 g, 13.1 mmol) is added to a solution of concentrated HCl (11.0 mL, 128.0 mmol) diluted with water (2 mL). A solution of NaNO2 (1 g, 14.5 mmol) in water (2 mL) is added dropwise to the mixture cooled to 0°C for 2 min, while maintaining the temperature at 0°C. After addition, the slurry is stirred at 0°C for 40 min to obtain a brown suspension. CuSO45H2O (3.6 g, 14.5 mmol) is added to HCl (10.6 mL, 127.5 mmol) and cooled to 0°C. NaHSO3 (2.7 g, 26.3 mmol) is dissolved in water (2 mL) and the solution is divided into two portions. The first portion is added to the hydrochloric acid solution. The brown suspension is added dropwise to the hydrochloric acid solution, while maintaining the temperature at 5°C. At the same time, another portion of NaHSO3 solution is added dropwise. The reaction mixture is then stirred at 0° C. for 1 h until no more gas is produced, completing the reaction. The reaction mixture is filtered and washed with DCM (300 mL). The combined organic layer is separated and dried over Na2SO4, filtered. The filtrate is concentrated under reduced pressure to give yellow oily compound 31-2 (3.2 g, 12.22 mmol, 93.0%).

[0272] To a solution of compound 23-6 (0.85 g, 3.72 mmol) and compound 31-2 (2.1 g, 8.19 mmol) in DCM (21 mL), DABCO (442 mg, 3.94 mmol) is added and the mixture is stirred at 25 °C for 16 h. The mixture is diluted with water (10 mL) and extracted with DCM (15 mL x 3). The organic phase is dried over MgSO4, filtered, and the filtrate is concentrated under vacuum. The residue is purified by flash silica gel chromatography (ISCO, 25 g SepaFlashSilica Flash Column, eluent 0-30%, ethyl acetate / petroleum ether gradient) to give compound 31-3 (1.83 g, 3.64 mmol, 97.7%) as a yellow solid. LC-MS(ESI):m / z 372.2[M-56] + ,

[0273] To a suspension of compound 31-3 (1.2 g, 2.39 mmol) in ethanol (25 mL) and water (25 mL) under nitrogen gas at 25°C, Fe (1.4 g, 25.1 mmol) and NH4Cl (1.3 g, 24.3 mmol) are added. The mixture is stirred at 90°C for 2 h. The mixture is a black suspension. The mixture is diluted with water (15 mL) and extracted with DCM (30 mL x 3). The combined organic phase is dried over MgSO4, filtered, and the filtrate is concentrated under vacuum to give compound 31-4 (1 g, 2.26 mmol, 94.7%) as a white solid. LC-MS(ESI):m / z 297.9[M-100] + ,

[0274] To a solution of compound 31-4 (80 mg, 0.20 mmol) and compound 31-5 (50 μL, 0.40 mmol) in acetonitrile (4 mL), add triethylamine (84 μL, 0.60 mmol) and T3P (50% in ethyl acetate, 0.6 mL, 1.0 mmol). Stir the reaction mixture at 50 °C for 16 h. Add sodium hydroxide (2 M aqueous solution, 2.0 mL, 4.0 mmol) to the mixture and stir the mixture at 20 °C for 2 h. Extract the mixture with ethyl acetate (10 mL × 2). Concentrate the combined organic phase. The residue was purified by preparative HPLC (column: Phenomenex Gemini C18 250×50 mm×10 um, mobile phase: [water (0.05% aqueous ammonia v / v)-acetonitrile], B%: 50%-70%, 10 min) to give compound 31-6 (63 mg, 0.12 mmol, 57.7%) as a white solid. LC-MS(ESI): m / z 416.2[M-100] + ,

[0275] To a solution of compound 31-6 (63 mg, 0.12 mmol) in DCM (1 mL) was added HCl / dioxane (4 M, 0.3 mL) and the mixture was stirred for 2 h at 20° C. The mixture was concentrated to give compound 207A (63.79 mg, 0.12 mmol, 99.8%) as a white solid.

[0276] The following compounds are synthesized according to the steps described in Example 31. JPEG0007653166000349.jpg236160JPEG0007653166000350.jpg246160JPEG0007653166000351.j pg246160JPEG0007653166000352.jpg246160JPEG0007653166000353.jpg246160JPEG00076531660 00354.jpg246160JPEG0007653166000355.jpg246160JPEG0007653166000356.jpg246160JPEG000 7653166000357.jpg246160JPEG0007653166000358.jpg246160JPEG0007653166000359.jpg112160

[0277] Example 32 JPEG0007653166000360.jpg4798To a solution of compound 126 (15 mg, 0.032 mmol), acetic acid (0.4 μL, 0.006 mmol), and formaldehyde (6.7 mg, 0.22 mmol) in methanol (1 mL), add 2-picoline borane (26.5 μL, 0.32 mmol) and stir the mixture at 80 °C for 16 h. Concentrate the mixture to obtain the crude product. Purify the residue by preparative HPLC (column: Phenomenex Gemini C18 250 × 50 mm × 10 um, mobile phase: water (0.05% ammonia water v / v)-acetonitrile, B%: 50%-70%, 10 min) to obtain white solid compound 214 (4.02 mg, 0.01 mmol, 26.2%).

[0278] The following compounds are synthesized according to the steps described in Example 32. JPEG0007653166000361.jpg236160JPEG0007653166000362.jpg225160JPEG0007653166000363.jpg245160JPE G0007653166000364.jpg245160JPEG0007653166000365.jpg245160JPEG0007653166000366.jpg245160JPEG000 7653166000367.jpg245160JPEG0007653166000368.jpg245160JPEG0007653166000369.jpg245160JPEG0007653 166000370.jpg245160JPEG0007653166000371.jpg234160JPEG0007653166000372.jpg245160JPEG00076531660 00373.jpg245160JPEG0007653166000374.jpg245160JPEG0007653166000375.jpg245160JPEG00076531660003 76.jpg245160JPEG0007653166000377.jpg244160JPEG0007653166000378.jpg245160JPEG0007653166000379.j pg245160JPEG0007653166000380.jpg246160JPEG0007653166000381.jpg246160JPEG0007653166000382.jpg24 6160JPEG0007653166000383.jpg235160JPEG0007653166000384.jpg246160JPEG0007653166000385.jpg193160

[0279] Example 33 JPEG0007653166000386.jpg66146

[0280] To a solution of compound 33-1 (50 g, 218.07 mmol) in N,N-dimethylformamide (300 mL) at 0 °C, add methoxy(methyl)amine hydrochloride (21.27 g, 218.07 mmol), 1-EDC.HCl (41.80 g, 218.07 mmol), DIEA (72.3 mL, 436.15 mmol) and HOBt (29.47 g, 218.07 mmol). Stir the reaction mixture overnight at room temperature. Dilute the mixture with water (1 L) and extract with ethyl acetate (1 L x 2). Wash the combined organic layer with brine (1 L), dry over Na2SO4, filter and concentrate. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate=100 / 1-0 / 1) to give colorless oily compound 33-2 (53.3 g, 195.70 mmol, yield: 89.7%). 1HNMR(400MHz,DMSO-d6)δ3.96(d,J=11.6Hz,2H),3.69(s,3H),3.09(s,3H),2.88-2.73(m,3H),1.64(d,J=12.0Hz,2H),1.40(s,9H).

[0281] Cyclopropylmagnesium bromide (27.5mL) is added to a solution of compound 33-2 (5g, 18.36mmol) in tetrahydrofuran (50mL) under 0°C. The reaction mixture is warmed to room temperature and stirred for 30 minutes, then diluted with saturated NaHCO3 solution (50mL). The mixture is extracted with ethyl acetate (50mL x 3). The combined organic layers are washed with brine (50mL), dried over Na2SO4, and concentrated in vacuum. The residue is purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 0 to 1 / 1) to give the title compound 33-3 (4g, 15.79mmol, yield: 86.0%) as a colorless liquid. 1HNMR(400MHz,DMSO-d6)δ3.91(d,J=12.4Hz,2H),2.90-2.71(m,3H),2.20-2.09(m,1H),1.8 2(d,J=12.6Hz,2H),1.39(s,9H),1.32(dd,J=12.3,2.8Hz,2H),0.83(dd,J=18.9,4.9Hz,4H).

[0282] To a solution of compound 33-3 (1 g, 3.95 mmol) in methanol (20 mL), NHOAC (6.09 g, 78.95 mmol) and sodium cyanoborohydride (2.48 g, 39.47 mmol) are added, and the reaction mixture is left overnight at 30° C. The mixture is quenched with saturated NHCl solution (50 mL) and extracted with ethyl acetate (50 mL×2). The combined organic phase is washed with brine (50 mL), dried over NaSO, filtered, and concentrated to give the crude product compound 33-4 (600 mg, 2.36 mmol) as a colorless oil. A mixture of compound 23-3 (328.8 mg, 1.06 mmol), compound 33-4 (200 mg, 0.78 mmol) and DABCO (357.2 mg, 3.18 mmol) in CHCl (10 mL) is stirred overnight at room temperature. The suspension is partitioned between DCM (20 mL) and water (20 mL). The aqueous layer is separated and extracted with CHCl (20 mL×2). The combined organic layer is washed with brine (20 mL), dried over anhydrous NaSO, filtered and concentrated. The residue is purified by silica gel column chromatography (petroleum ether / ethyl acetate=10 / 1-1 / 1, DCM / methanol=60 / 1) to give compound 33-5 (282 mg, 0.53 mmol, 50.3% yield) as a white amorphous solid.

[0283] To a mixture of compound 225A (160 mg, 0.32 mmol), formaldehyde (67.8 mg, 2.26 mmol) and acetic acid (3.87 mg, 0.065 mmol) in methanol (10 mL) is added 2-picoline borane (345.00 mg, 3.23 mmol). The reaction mixture is stirred at 75° C. for 2 h. The solvent is removed and the residue is purified by preparative HPLC (NH3H2O / ACN / H2O) to give compound 226 (50.98 mg, 0.11 mmol, 33.4% yield) as a white solid.

[0284] The following compounds are synthesized according to the steps described in Example 33. JPEG0007653166000387.jpg236160JPEG0007653166000388.jpg246160JPEG00076531660 00389.jpg225160JPEG0007653166000390.jpg245160JPEG0007653166000391.jpg112160

[0285] Example 34 JPEG0007653166000392.jpg5483

[0286] To a solution of compound 216 (60 mg, 0.14 mmol) in tetrahydrofuran (3 mL), LiAlH4 (15.5 mg, 0.42 mmol) is added, and the reaction mixture is stirred at 66 ° C for 3 h. The mixture is cooled to room temperature, and water (0.1 mL) is added, dried over Na2SO4, filtered, and the filtrate is concentrated under vacuum. The residue is purified by preparative HPLC (system: Waters 2767 / 2545 / 2489 / Qda, column name: Waters Xbridge C18 10 um OBD 19 × 150 mm, mobile phase A: 10 mM NH4OH aqueous solution, mobile phase B: acetonitrile, wavelength: 254 nm / 214 nm, flow rate: 20 mL / min: column temperature: room temperature) to obtain white solid compound 229 (2.09 mg, 0.005 mmol, yield: 3.6%).

[0287] JPEG0007653166000393.jpg188160

[0288] Example 35 JPEG0007653166000394.jpg58107

[0289] To a solution of compound 161A (150 mg, 0.33 mmol) in N,N-dimethylformamide (8 mL), add triethylamine (0.18 mL, 1.33 mmol) and stir the mixture at room temperature for 10 minutes, then add a solution of acetyl chloride (26 mg, 0.33 mmol) in tetrahydrofuran (1 mL). Stir the reaction mixture at room temperature for 1 hour and dilute with DCM (30 mL) and saturated sodium chloride solution (20 mL). Separate the organic layer, dry with Na2SO4, and concentrate under vacuum. Purify the residue by silica gel column chromatography (methanol / DCM=0 / 1-1 / 20) to obtain the title compound 230 (60 mg, 0.13 mmol, yield: 39.5%) as a white solid.

[0290] JPEG0007653166000395.jpg236160JPEG0007653166000396.jpg58160

[0291] Example 36 JPEG0007653166000397.jpg33145

[0292] To a solution of compound 23-6 (240 mg, 0.89 mmol) in DCM (5 mL), compound 36-1 (203.3 mg, 0.89 mmol) and triethylamine (299.6 mg, 2.67 mmol) are added. The reaction mixture is stirred at room temperature for 1 h and concentrated in vacuum. The residue is purified by silica gel column chromatography (petroleum ether / ethyl acetate: 100 / 0-20 / 1) to give compound 36-2 (385 mg, 0.83 mmol, yield: 93.7%). LC-MS(ESI):m / z 361.9[M+H-Boc] +

[0293] At room temperature, cesium carbonate (360.12 mg, 1.11 mmol) and Pd2(dba)3 (9.92 mg, 0.01 mmol), X-PHOS (35.07 mg, 0.07 mmol), methylamine (0.07 mL, 1.84 mmol) are added to a solution of compound 36-2 (220 mg, 0.477 mmol) in dioxane (2 mL), and the reaction mixture is stirred at 110° C. for 4 hours under nitrogen gas atmosphere. The mixture is cooled to room temperature and concentrated. The residue is purified by silica gel column chromatography (petroleum ether / ethyl acetate: 100 / 1-10 / 1) to give compound 36-3 (100 mg, 0.24 mmol, yield: 66.0%). LC-MS(ESI): m / z 312.2[M+H-Boc] +

[0294] To a solution of compound 36-3 (100 mg, 0.24 mmol) in toluene (5 mL), 2-methylbenzoyl (70 μL, 0.55 mmol) and Et3N (140.14 mg, 1.39 mmol) are added. The reaction mixture is stirred at 100° C. for 5 h. The mixture is cooled to room temperature and concentrated. The residue is purified by silica gel column chromatography (petroleum ether / ethyl acetate: 100 / 1-10 / 1) to give compound 36-4 (100 mg, 0.36 mmol, yield: 77.7%). LC-MS(ESI): m / z 430.1[M+H-Boc] +

[0295] To a flask containing compound 231A (137 mg, 0.26 mmol), add HCl / dioxane (4N, 5 mL). The mixture is stirred at room temperature for 1 hour and concentrated. The residue is purified by preparative HPLC (system: Waters 2767 / 2545 / 2489 / Qda, column name: Waters Xbridge C18 10 um OBD 19×150 mm, mobile phase A: 10 mM hydrochloric acid aqueous solution, mobile phase B: acetonitrile, wavelength: 254 nm / 214 nm, flow rate: 20 mL / min: column temperature: room temperature) to obtain IMP-7282 (13.58 mg, 0.03 mmol, yield: 12.2%).

[0296] JPEG0007653166000398.jpg236160JPEG0007653166000399.jpg227160

[0297] Example 37 JPEG0007653166000400.jpg46114

[0298] To a solution of compound 37-1 (202 mg, 0.62 mmol) in DCM (5 mL), compound 23-3 (100 mg, 0.63 mmol) and triethylamine (211 mg, 1.88 mmol) are added and the reaction mixture is stirred at room temperature for 2 h. The mixture is concentrated and the residue is purified by preparative HPLC (system: Waters 2767 / 2545 / 2489 / Qda, column name: Waters Xbridge C18 10 um OBD 19 × 150 mm, mobile phase A: 10 mM NH4OH in water, mobile phase B: acetonitrile, wavelength: 254 nm / 214 nm, flow rate: 20 mL / min, column temperature: room temperature) to obtain IMP-7300-A (48 mg, 0.11 mmol, yield: 16.0%).

[0299] Lithium aluminum hydride (30.6 mg, 0.81 mmol) is added to a solution of compound 232 (120 mg, 0.27 mmol) in tetrahydrofuran (5 mL) under N2 at -78°C, the reaction mixture is stirred at 0°C for 3 hours, and then tetrahydrofuran (5 mL) and water (0.036 mL) are added. The mixture is dried over Na2SO4, the organic layer is separated and concentrated. The residue is purified by preparative HPLC (system: Waters 2767 / 2545 / 2489 / Qda, column name: Waters Xbridge C18 10um OBD19×150mm, mobile phase A: 10 mM NH4OH aqueous solution, mobile phase B: acetonitrile, wavelength: 254 nm / 214 nm, flow rate: 20 mL / min, column temperature: room temperature) to obtain compound 233 (23.23 mg, 0.06 mmol, yield: 21.4%).

[0300] JPEG0007653166000401.jpg236160JPEG0007653166000402.jpg239160

[0301] Example 38 JPEG0007653166000403.jpg50141

[0302] Compound 38-1 (40μL, 0.36mmol) is added to a solution of compound 31-4 (97.9mg, 0.24mmol) and triethylamine (0.10mL, 0.73mmol) in toluene (15mL), and the reaction mixture is stirred at 120℃ for 2h. The mixture is cooled to room temperature, diluted with water (100mL), and extracted with ethyl acetate (50mL×3). The combined organic layer is washed with brine (50mL×3), dried over anhydrous Na2SO4, filtered and concentrated. The residue is purified by silica gel column chromatography (ethyl acetate / petroleum ether=1 / 100-1 / 1) to give compound 38-2 (78mg, 0.13mmol, 52.7% yield) as a white solid. LC-MS(ESI): m / z 517.2[M+H] +

[0303] To a solution of compound 38-2 (78 mg, 0.15 mmol) in 1,4-dioxane (2 mL) is added HCl(g) in dioxane (4N, 2 mL). The reaction mixture is stirred at room temperature for 2 h and concentrated in vacuum. The residue is purified by preparative HPLC (acidic method: HCl in CH3CN) to give compound 236A (22.99 mg, 0.05 mmol, 32.6% yield) as a white solid.

[0304] JPEG0007653166000404.jpg181160

[0305] Example 39 JPEG0007653166000405.jpg77146

[0306] To a solution of compound 23-6 (145.4 mg, 0.64 mmol) and DABCO (214.2 mg, 1.91 mmol) in DCM (10 mL) is added compound 31-2 (150 mg, 0.64 mmol). The reaction mixture is stirred at room temperature for 2 h and diluted with water (50 mL). The mixture is extracted with DCM (100 mL x 2). The combined organic layer is washed with brine (50 mL x 3), dried over anhydrous Na2SO4, filtered and concentrated. The residue is purified by silica gel column (ethyl acetate / petroleum ether = 1 / 20-1 / 2) to give compound 39-1 (219 mg, 0.51 mmol, 80.5% yield) as a white solid. LC-MS(ESI): m / z 426.6[MH] - ;

[0307] At 0°C, sodium hydride (26.6mg, 0.67mmol) is added to a solution of compound 39-1 (219mg, 0.51mmol) in N,N-dimethylformamide (10mL) and stirred at room temperature for 30 minutes, then iodomethane (0.16mL, 2.561mmol) is added. The reaction mixture is stirred at room temperature for 30 minutes and diluted with ethyl acetate (50mL) and brine (50mL). The organic layer is separated, washed with brine (50mL), dried over Na2SO4, and concentrated under vacuum. The crude product is purified by silica gel column (ethyl acetate / petroleum ether=1 / 100-1 / 10) to obtain yellow solid compound 39-4 (291mg, 0.66mmol, 128.7% yield). LC-MS(ESI):m / z 342.0[M-Boc+H] + .

[0308] To a solution of compound 39-2 (291 mg, 0.66 mmol) in methanol (10 mL), Pd / C (70.1 mg, 0.66 mmol) is added and the mixture is degassed with hydrogen gas three times. The reaction mixture is stirred at 30° C. for 5 h. The Pd / C is filtered off. The filtrate is concentrated to give compound 39-3 (175 mg, 0.43 mmol, 64.5% yield, Lot#: N190849-120-P1) as a white amorphous solid. LC-MS(ESI): m / z 411.1[MH]-

[0309] To a solution of compound 39-3 (175 mg, 0.43 mmol) in DCM (10 mL) and triethylamine (0.18 mL, 1.28 mmol), a solution of compound 23-4 (0.06 mL, 0.47 mmol) in DCM (5 mL) is added. The reaction mixture is stirred at room temperature for 30 min, and then water (50 mL) is added. The mixture is extracted with DCM (100 mL x 2). The combined organic layer is washed with brine (50 mL x 3), dried over anhydrous Na2SO4, filtered and concentrated. The residue is purified by silica gel column (ethyl acetate / petroleum ether = 1 / 100-1 / 10) to give compound 39-4 (138 mg, 0.26 mmol, 61.3% yield) as a white solid. LC-MS(ESI): m / z 529.1[MH] +

[0310] To a solution of compound 39-4 (138 mg, 0.26 mmol) in 1,4-dioxane (5 mL), add HCl (g) in dioxane (4N, 5 mL). Stir the reaction mixture at room temperature for 30 min. Add DCM (20 mL) and concentrate. Purify the residue by preparative HPLC (acidic method: Waters 2767 / 2545 / 2489, Inertsil ODS-310um 20×250 nm, mobile phase A: 0.1% aqueous hydrochloric acid, mobile phase B: acetonitrile, flow rate: 20 mL / min, column temperature: room temperature) and dry in the cold to obtain compound 237A (47.01 mg, 0.10 mmol, 38.6% yield) as a white solid.

[0311] JPEG0007653166000406.jpg210160

[0312] CCR8 Inhibition in Vitro Studies The biological properties of the new compounds are studied based on the following in vitro measurement methods:

[0313] Tango analysis The Tango cell line is used to detect antagonist-mediated receptor inactivation. Tango cells (Tango-CCR8-Gal4-CHO-K1 cells constructed by Genomeditech) are cultured in complete medium of F12K, 10% fetal bovine serum, 1% penicillin-streptomycin, 4 μg / ml puromycin, 4 μg / ml blasticidin and 100 μg / ml hygromycin in an incubator (37°C, 5% carbon dioxide) for subculture. 24 h prior to cell plating, culture the cells in 10 cm or 15 cm culture dishes at 30% confluence in medium (F12K, 10% fetal bovine serum, 1% penicillin-streptomycin).

[0314] On the day of plating, cells are digested with 0.25% trypsin / ethylenediaminetetraacetic acid (EDTA) and counted. Cells are suspended in starvation medium (F12K, 1% fetal bovine serum, 1% penicillin-streptomycin) and then plated in black / clear 96-well plates (1×10 4 Cells / 70μl / well) and incubated for 6 hours (37℃, 5% carbon dioxide). Compounds at different concentrations were dissolved in DMSO and added to a 96-well plate at 50μL per well and incubated for 1 hour at 37℃. Human CCL1 protein (R&D, Product No.: 272-I) dissolved in starvation medium (F12K, 1% fetal bovine serum, 1% penicillin-streptomycin) was added to a 96-well plate at 30μL per well and incubated for 24 hours at 37℃. After incubation, remove the medium and place the 96-well plate in a -80°C refrigerator for 1 hour or more. Remove the plate and ONE-Glo Reagent (Promega, product number: E6110) frozen at -80°C and bring to room temperature. ONE-Glo Working Reagent is prepared by mixing ONE-Glo Reagent with F12K (no fetal bovine serum) (1:2), and add 40 μl of ONE-Glo Working Reagent to each well. After incubating at room temperature for 10 minutes, read the fluorescence value at 560 nm wavelength using a microplate reader.

[0315] The microplate reader records fluorescence values ​​in relative luminescence units (RLU). A single excel file containing the RLU data is exported and the RLU results are plotted against various drug concentrations and analyzed in GraphPad Prism 7.0 to generate concentration curves. The Tango analysis results IC50 data of the compounds for CCR8 inhibitory effect are shown in Table 1 below.

[0316] JPEG0007653166000407.jpg228160JPEG0007653166000408.jpg244160JPEG0007653166000409.jpg245160JPEG0007653166000410.jpg244160JPEG000 7653166000411.jpg244160JPEG0007653166000412.jpg244160JPEG0007653 166000413.jpg244160JPEG0007653166000414.jpg244160JPEG00076531660 00415.jpg244160JPEG0007653166000416.jpg244160JPEG0007653166000417.jpg244160JPEG0007653166000418.jpg244160JPEG0007653166000419.j pg244160JPEG0007653166000420.jpg244160JPEG0007653166000421.jpg244160JPEG0007653166000422.jpg244160JPEG0007653166000423.jpg217160

[0317] Ca 2+ Mobility measurement Calcium ion mobilization assay is a cell-based second messenger assay to measure calcium flux associated with activation or inhibition of G protein-coupled receptors. The change in fluorescence intensity is directly related to the content of intracellular calcium released into the cytoplasm in response to ligand activation of the target receptor. A membrane-permeable calcium-binding fluorescent dye (Fluo4 AM, Solarbio F8500) was dissolved in anhydrous dimethyl sulfoxide (DMSO) to prepare a 2 mM stock solution, which was also supplemented with Pluronic F127 (0.01%–0.02%) and stored in aliquots at -80°C in the dark.

[0318] HEK293 cells were suspended in standard medium at approximately 2.5 × 10 5 The cells are seeded in 6-well plates at 1000 cells / well and cultured for 2 days (approximately 70% confluence), after which Fluo4 AM (final concentration is 4 μM) is added. The cells are washed with DMEM, followed by adding 1 mL of loading dye mixture to each well and incubating for 30 minutes in a humidified incubator at 37°C and 5% CO2, protected from light. Aspirate the calcium dye solution from each well, wash the cells with Duchenne phosphate buffered saline (D-PBS) containing 0.25% trypsin / ethylenediaminetetraacetic acid (EDTA), and continue to incubate at 37°C until the cells are completely detached. Resuspend the cells in complete medium and centrifuge at 200 g RPM for 3 minutes in a microcentrifuge. Carefully aspirate the supernatant and resuspend the pellet in DMEM. After repeating the wash twice, resuspend the pellet in DMEM in a final volume of 500 μl. Calcium ion mobility is measured at room temperature. Briefly, the cell suspension is placed in a 1.5 mL Eppendorf tube. After incubation for 1 min for baseline fluorescence, test compounds are added; this phase should not exceed 15 s. The calcium signal is recorded for 3-4 min. Ionomycin (10 μM) and ethylene glycol tetraacetic acid (EGTA) are used as positive and negative controls, respectively. Ca of compounds on CCR8 inhibitory effect 2+ Mobility measurement result IC 50 The data is shown in Table 2 below.

[0319] JPEG0007653166000424.jpg231160JPEG0007653166000425.jpg51160

[0320] While the present invention has been particularly shown and described with reference to preferred embodiments thereof, it will be understood by those skilled in the art that various changes in form and detail can be made therein without departing from the scope of the invention contained in the appended claims.

Claims

1. A compound selected from the following: or a pharma- ceutically acceptable salt, isotope, stereoisomer, or tautomer thereof:

2. The compound is 2. The compound of claim 1, wherein:

3. The compound is 2. The compound of claim 1, wherein:

4. The compound is 2. The compound of claim 1, wherein:

5. 1. A pharmaceutical composition for ameliorating disease symptoms in a subject having a disease mediated by an abnormal or undesirable CCR8 / CCL1 axis, comprising: A pharmaceutical composition comprising a compound according to any one of claims 1 to 4 or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable excipient; The pharmaceutical composition, wherein the disease is selected from the group consisting of cancer and neuropathic pain.

6. The pharmaceutical composition of claim 5 , wherein the cancer comprises leukemia or a solid cancer.

7. The leukemia includes chronic lymphocytic leukemia, chronic myelogenous leukemia, acute myelogenous leukemia, acute lymphoblastic leukemia and lymphoma leukemia stage; The pharmaceutical composition of claim 6, wherein the solid tumor comprises breast cancer, gastric cancer, colorectal cancer, ovarian cancer, pancreatic cancer and liver cancer.

8. The pharmaceutical composition according to claim 5 , wherein the neuropathic pain is caused by diabetes or spinal cord injury.

Images