阅读说明：本技术 免疫调节剂及其组合物和制备方法 (Immunomodulator, composition and preparation method thereof ) 是由 张垚 王义乾 付邦 陈洁 王家炳 丁列明 于 2019-07-19 设计创作，主要内容包括：本发明涉及式Ⅰ化合物,使用该化合物作为免疫调节剂的方法,包括该化合物的药用组合物。该化合物可用于治疗,预防或改善疾病或病症,例如癌症或感染。(The present invention relates to compounds of formula (I), methods of using the compounds as immunomodulators, pharmaceutical compositions comprising the compounds. The compounds are useful for treating, preventing or ameliorating a disease or disorder, such as cancer or infection.)

1. A compound of formula I, or a stereoisomer, tautomer, pharmaceutically acceptable salt, prodrug, chelate, non-covalent complex, or solvate thereof,

wherein the content of the first and second substances,

ring A and ring B are each independently selected from a 5-to 6-membered heterocyclic ring; the heterocycle optionally comprises 1, 2 or 3 heteroatoms independently selected from N, S, or O;

represents a single or double bond including cis-trans isomers;

if it is notRepresents a double bond, X and Y are each independently selected from- (CH)_n-; if it is notRepresents a single bond, X and Y are each independently selected from absent, - (CH)₂)_n-，-S-，-O-，-NR₈-，-CO-，-CONR₉-, or-NR₁₀CO-，-SO₂- (CH)₂)_n-，-NR₈-，-CONR₉-, or-NR₁₀CO-optionally substituted by C_1-8Alkyl, -OC_1-8Alkyl substituted;

R₁，R₂and R₃Each independently selected from H, halogen, CN, C_1-8Alkyl radical, -C_1-8Haloalkyl, or-OC_1-8Alkyl groups: or

R₁And X together with the atoms to which they are attached form a 5-to 6-membered heterocyclic ring; the heterocycle optionally comprises 1, 2 or 3 heteroatoms independently selected from N, S, or O; said heterocycle being optionally substituted by C_1-8Alkyl radical, -C_0-4alkyl-COOH, or-C_0-4alkyl-OH; or

R₃And Y together with the atoms to which they are attached form a 5-to 6-membered heterocyclic ring; the heterocycle optionally comprises 1, 2 or 3 heteroatoms independently selected from N, S, or O; said heterocycle being optionally substituted by C_1-8Alkyl radical, -C_0-4alkyl-COOH, or-C_0-4alkyl-OH;

R₄and R₅Each independently selected from H, -CONH₂，-C_1-8Alkyl radical, -C_1-8Alkenyl, -C_1-8Haloalkyl, -C_1-8Heteroalkyl group, C_3-10Cycloalkyl, -C_1-4alkyl-C_5-6Aryl, -CO-C_1-4Alkyl, -SO₂-C_1-4Alkyl radical, -C_1-4alkyl-COOH, -C_1-4alkyl-OH; or

R₄And R₅Form a 5-to 6-membered heterocyclic ring with the atom to which they are attached; the heterocycle optionally comprises 1, 2 or 3 heteroatoms independently selected from N, S, or O; said heterocycle being optionally substituted by C_1-8Alkyl radical, -C_0-4alkyl-COOH, or-C_0-4alkyl-OH;

R₆and R₇Each independently selected from H, -C_1-8Alkyl radical, -C_1-8Heteroalkyl, or C_3-10Cycloalkyl radicals in which-C_1-8Alkyl radical, -C_1-8HeteroalkanesRadical, or C_3-10Cycloalkyl is optionally substituted with-COOH or-OH; or

R₆And R₇Form a 4-to 6-membered heterocyclic ring with the atom to which they are attached; the heterocycle optionally comprises 1, 2 or 3 heteroatoms independently selected from N, S, or O; said heterocycle being optionally substituted by-C_0-4alkyl-COOH, or-C_0-4alkyl-OH;

R₈，R₉，R₁₀each independently selected from H, -C_1-4Alkyl radical, -C_1-4Haloalkyl, -C_1-8Heteroalkyl group, -C_1-4alkyl-COOH, or-C_1-4alkyl-OH;

R₁₁and R₂₂Each independently selected from H, halogen, CN, or C_1-8An alkyl group; or

R₁₁And R₂₂Form a 3-to 4-membered heterocyclic ring with the atom to which they are attached;

n is 1, 2 or 3.

2. The compound of claim 1, wherein ring a is a 6-membered heterocyclic ring comprising 1, 2, or 3 heteroatoms independently selected from N, or S.

3. The compound of claim 1 or claim 2, wherein ring B is a 6-membered heterocyclic ring comprising 1, 2, or 3 heteroatoms independently selected from N, or S.

4. A compound as claimed in claims 1-3 whereinIs composed of

5. The compound of claims 1-4, wherein R₁And R₂Each independently selected from H, F, Cl, CN, or methyl.

6. The compound of claims 1-5, wherein R₃Is H, F, Cl, CH₃Or CF₃。

7. The compound of claims 1-6, wherein R₄And R₅Form a 5-to 6-membered heterocyclic ring with the atoms to which they are attached.

8. The compound of claim 7, wherein the 5-to 6-membered heterocyclic ring is

9. The compound of claim 8 wherein the 5-to 6-membered heterocycle is optionally substituted with-COOH or-CH₃And (4) substituting.

10. The compound of claims 1-6, wherein R₄And R₅Each independently selected from H, C_1-4Alkyl, -CO-C_1-4Alkyl, -SO₂-C_1-4Alkyl radical, -C_1-4alkyl-COOH, -C_1-4alkyl-OH.

11. The compound of claims 1-10, wherein R₆And R₇Form a 6-membered heterocyclic ring with the atom to which they are attached.

12. The compound of claim 11, wherein the heterocycle is substituted with-COOH.

13. The compound of claim 12, wherein R₈，R₉，R₁₀Each independently selected from H, or methyl.

14. The compound of claims 1-13, R₁₁And R₂₂Each independently selected from H, or methyl.

15.The compound of claims 1-13, R₁₁And R₂₂With the atoms to which they are attached constituting a 3-membered carbocyclic ring.

16. The compound of any one of claims 1-14, n is 1.

17. A compound of formula I which is

1) ((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) glycine;

2) ((8- ((2-methyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) glycine;

3) ((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) proline;

4) ((8- ((2-methyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) proline;

5)1- ((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-2-yl) methyl) piperidine-2-acetic acid;

6)1- ((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) -4-hydroxypyrrolidine-2-acetic acid;

7)3- (((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) amino) propanoic acid;

8)4- (((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) amino) butanoic acid;

9)1- ((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) pyrrolidine-3-acetic acid;

10) ((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) serine;

11) n- ((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) -N-methylglycine;

12)1- ((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) -4, 4-difluoropyrrolidine-2-acetic acid;

13)2- (((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) amino) -3-hydroxy-2-methylpropionic acid;

14) n- ((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) -N-ethylglycine;

15)1- ((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) azetidine-3-acetic acid;

16)1- ((8- ((2,2' -dimethyl-3 ' - (3- (piperidin-1-yl) propoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

17)1- ((8- ((2,2' -dimethyl-3 ' - (3- (4-methylpiperazin-1-yl) propoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

18)1- ((8- ((2,2' -dimethyl-3 ' - (3- (pyrrolidin-1-yl) propoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

19)1- ((8- ((3' - (3- (3-hydroxypyrrolidin-1-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

20)1- ((8- ((2-methyl-3- (1- (3-morpholinopropyl) indolin-4-yl) phenyl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

21)1- ((8- ((3' - (3- (diethylamino) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

22)1- ((8- ((3' - (3- (benzyl (methyl) amino) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

23)1- ((8- ((3' - (3- (ethylamino) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

24)1- ((8- ((3' - (3-acetylaminopropoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

25)1- ((8- ((2,2' -dimethyl-3 ' - (3-ureidopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

26)1- ((8- ((3' - (3-guanidinopropoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

27)1- ((8- ((2,2' -dimethyl-3 ' - (3- (methylsulfonylamino) propoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

28)1- ((8- ((3' - (3- ((carboxymethyl) amino) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

29)1- ((8- ((3' - (3- (2-carboxypyrrolidin-1-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

30)1- ((8- ((3' - (3- (4-carboxypiperidin-1-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

31)1- ((8- ((2,2' -dimethyl-3 ' - (2-morpholinoethoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

32)1- ((8- ((2-methyl-3- (7- (2-morpholinoethoxy) naphthalen-2-yl) phenyl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

33)1- ((8- ((2-methyl-3 '- (3- (oxetan-3-ylamino) propoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

34)1- ((8- ((2-methyl-3- (1- (3-morpholinopropyl) -1H-indol-4-yl) phenyl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

35)1- ((8- ((2-methyl-3- (2- (2-morpholinoethyl) -1H-indol-6-yl) phenyl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

36)1- ((8- ((2-methyl-3 '- (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

37)1- ((8- ((2' -fluoro-2-methyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

38)1- ((8- ((2' -chloro-2-methyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

39)1- ((8- ((2' -cyano-2-methyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

40)1- ((8- ((4' -fluoro-2-methyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

41)1- ((8- ((2,2',4' -trimethyl-3 '- (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

42)1- ((8- ((2,4' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

43)1- ((8- ((4' -chloro-2-methyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

44) ((8- ((4' -chloro-2-methyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) glycine;

45) ((8- ((2' -chloro-2-methyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) glycine;

46) ((8- ((2-methyl-3- (4-methyl-5- (3-morpholinopropoxy) pyridin-3-yl) phenyl) amino) -1, 7-naphthyridin-3-yl) methyl) glycine;

47)1- ((8- ((2,2' -dimethyl-3 ' - ((2-morpholinoethoxy) methyl) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

48)1- ((8- ((2,2' -dimethyl-3 ' - ((3-morpholinopropyl) amino) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

49)1- ((8- ((2,2' -dimethyl-3 ' - (3-morpholinepropionamide) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

50)1- ((8- ((2,2' -dimethyl-3 ' - (N-methyl-3-morpholinepropionamide) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

51)1- ((8- ((3' - (3- (2- ((2-hydroxyethyl) amino) ethyl) urea) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

52)1- ((8- ((2,2' -dimethyl-3 ' - (methyl (3-morpholinopropyl) amino) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

53)1- ((8- ((2,2' -dimethyl-3 ' - (3- (2-morpholinoethyl) urea) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

54)1- ((8- ((2,2' -dimethyl-3 ' - ((1- (morpholinomethyl) cyclopropyl) methoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

55)1- ((8- ((2,2' -dimethyl-3 ' - ((4-morpholinebutan-2-yl) oxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

56)1- ((8- ((2-methyl-3- (1- (2-morpholinoethoxy) -2, 3-dihydro-1H-indan-4-yl) phenyl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

57) (E) -1- ((8- ((2,2' -dimethyl-3 ' - (4-morpholinobutyl-1-en-1-yl) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

58)1- ((8- ((2,2' -dimethyl-3 ' - (4-morpholinoethyl) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

59)1- ((8- ((4' -methoxy-2-methyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

60)1- ((8- ((2-methyl-3 ' - (3-morpholinopropoxy) -4' - (trifluoromethyl) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid.

18. A pharmaceutical composition comprising a compound of any one of claims 1-17, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, and at least one pharmaceutically acceptable carrier or excipient.

19. A method of inhibiting PD-1/PD-L1 binding, the method comprising administering to a patient a compound of any one of claims 1-17, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof.

20. A method of treating a disease associated with inhibition of PD-1/PD-L1 binding, the method comprising administering to a patient in need thereof a therapeutically effective amount of a compound of any one of claims 1-17, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof.

21. The method of claim 20, wherein the disease is colon cancer, gastric cancer, thyroid cancer, lung cancer, leukemia, pancreatic cancer, melanoma, multiple melanoma, brain cancer, renal cancer, prostate cancer, ovarian cancer, or breast cancer.

22. A method of promoting, stimulating or increasing an immune response in a patient, comprising administering to a patient in need thereof a therapeutically effective amount of a compound of any one of claims 1-17, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof.

23. Use of a pharmaceutical composition according to claim 18 or a compound according to any one of claims 1 to 17 in the manufacture of a medicament.

24. The use according to claim 23, for the treatment or prevention of cancer.

25. The use according to claim 24, wherein the cancer is colon, stomach, thyroid, lung, leukemia, pancreatic, melanoma, multiple melanoma, brain, kidney, prostate, ovarian or breast cancer.

26. The use according to claim 23, as an inhibitor of PD-1/PD-L1 binding.

Technical Field

The present invention relates to pharmaceutically active compounds. The invention provides the compound, and a composition and an application method thereof. The compounds modulate PD-1/PD-L1 protein/protein interactions and are useful for the treatment of a variety of diseases including infectious diseases and cancer.

Background

The immune system plays an important role in controlling and eliminating diseases such as cancer. However, cancer cells often escape or suppress the immune system by some strategy, thereby promoting their growth. One of these is the alteration of the expression of costimulatory and costimulatory molecules on immune cells (Postowetal, J.clinical Oncology 2015, 1-9). Blocking the signals of inhibitory immune checkpoints such as PD-1 has proven to be a promising and effective therapeutic approach.

The interaction between PD-1 and PD-L1 leads to a reduction in tumor infiltrating lymphocytes, a reduction in T cell receptor-mediated cell proliferation and immune escape of Cancer cells (Dong et al, J.mol Med., 81:281-287 (2003); Blank et al, Cancer Immunol Immunother., 54:307-314 (2005); Konishi et al, Clin.cancer Res. 10:5094-5100 (2004)). This immunosuppressive effect can be reversed by blocking the local interaction of PD-1 with PD-L1 and is more pronounced when the interaction of PD-1 with PD-L2 is blocked (Iwai et al, Proc. Natl. Acad. Sci. USA, 99:12293-12297 (2002); Brown et al, J. Immunol, 170:1257-1266 (2003)).

Programmed death receptor 1, also known as CD279, is a cell surface receptor expressed on activated T cells, natural killer cells, B cells and macrophages (Greenwald et al, Annu. Rev. Immunol 2005, 23: 515-Asn 548; Okazaki and Honjo, Trends Immunol 2006, (4): 195-Asn 201). Has the function of negative feedback regulation system, and can prevent the activation of T cells to reduce autoimmunity and enhance self tolerance. In addition, PD-1 is also known to play a key role in inhibiting antigen-specific T cell responses in diseases such as cancer and viral infections. (Sharpe et al, Nat Immunol 20078, 239-.

PD-1 consists of an extracellular immunoglobulin variable-like domain, a transmembrane region and an intracellular domain (Parry et al, Mol Cell Biol 2005, 9543-9553). The intracellular domain contains two phosphorylation sites located in an immunoreceptor tyrosine-based inhibitory motif and an immunoreceptor tyrosine-based switching motif, suggesting that PD-1 negatively regulates T-cell receptor-mediated signaling. PD-1 has two ligands, PD-L1 and PD-L2(Parry et al, Mol Cell Biol 2005, 9543-9553; Latchman et al, Nat Immunol 2001, 2, 261-268), which are expressed differently. PD-L1 protein is up-regulated in macrophage and dendritic cells following lipopolysaccharide and GM-CSF treatment, and in T cells and B cells following T cell receptor and B cell receptor signaling. PD-L1 is highly expressed in almost all tumor cells and expression is further increased after IFN- γ treatment (Iwai et al, PNAS2002, 99 (19): 12293-7; Blank et al, Cancer Res 2004, 64 (3): 1140-5). Indeed, tumor PD-L1 expression status has been shown to be prognostic in a variety of tumor types (Wang et al, Eur J Surg Oncol 2015; Huang et al, Oncol Rep 2015; Sabatier et al, Oncotarget 2015, 6 (7): 5449-5464). In contrast, expression of PD-L2 is more restricted and is expressed predominantly by dendritic cells (Nakae et al, J Immunol 2006, 177: 566-73). The attachment of PD-1 and its ligands PD-L1 and PD-L2 to T cells can produce relevant signals to inhibit IL-2 and IFN-. gamma.production and cell proliferation induced by T cell receptor activation (Carter et al, Eur J Immunol 2002, 32 (3): 634-43; Freeman et al, J Exp Med 2000, 192 (7): 1027-34). This mechanism involves the recruitment of either SHP-2 or SHP-1 phosphatases to inhibit T cell receptor signaling such as phosphorylation of Syk and Lck (Sharpe et al, Nat Immunol 2007, 8, 239-245). Activation of the PD-1 signaling axis also attenuates phosphorylation of the PKC-theta activation loop, which is essential for activation of the NF-. kappa.B and API pathways and production of cytokines such as IL-2, IFN-. gamma.and TNF (Sharpe et al, Nat Immunol 2007, 8, 239-.

Some evidence from preclinical animal studies suggests that PD-1 and its ligands may have a negative regulatory effect on the immune response. PD-1 knockout mice develop lupus-like glomerulonephritis and dilated cardiomyopathy (Nishimura et al, Immunity 1999, 11: 41-151; Nishimura et al, Science 2001, 291: 319-. In a model of chronic LCMV viral infection, the PD-1/PD-L1 interaction has been shown to inhibit activation, expansion and acquisition of effector functions of virus-specific CD 8T cells (Barber et al, Nature 2006, 439, 682-7)).

These data support us to develop a therapeutic approach to enhance or "rescue" T cell responses by blocking the PD-1 mediated inhibitory signaling cascade. Most drugs currently approved in immunotherapy are monoclonal antibodies. However, small molecule inhibitors that directly target PD-1 or PD-L1 have not been approved, and only CA170 has been clinically evaluated.

There is therefore still a strong need for more effective and easier to administer therapeutic drugs directed to the PD-1 and PD-L1 protein/protein interactions. In the present invention, applicants have discovered that a potent small molecule can act as an inhibitor of the interaction of PD-L1 with PD-1 and therefore can be used in therapeutic administration to enhance immunity against cancer and/or infectious diseases. These small molecules are expected to be drugs with good stability, solubility, bioavailability, therapeutic index and toxicity values, which are important for being effective drugs for promoting human health.

Disclosure of Invention

The present invention relates to compounds useful as inhibitors of the interaction of PD-L1 and PD-1. Inhibitors of the interaction of PD-1 and PD-L1 may be useful in the treatment of cancer and other infectious diseases.

The compounds of the invention have the general structure shown in formula I. A compound of formula I, or a stereoisomer, tautomer, pharmaceutically acceptable salt, prodrug, chelate, non-covalent complex or solvate thereof,

wherein the content of the first and second substances,

ring A and ring B are each independently selected from a 5-to 6-membered heterocyclic ring; said heterocycle optionally comprises 1, 2 or 3 heteroatoms independently selected from N, S or O;

represents a single or double bond including cis-trans isomers;

if it is notRepresents a double bond, X and Y are each independently selected from- (CH)_n-; if it is notRepresents a single bond, X and Y are each independently selected from absent, - (CH)₂)_n-，-S-，-O-，-NR₈-，-CO-，-CONR₉-, or-NR₁₀CO-，-SO₂- (CH)₂)_n-，-NR₈-，-CONR₉-, or-NR₁₀CO-optionally substituted by C_1-8Alkyl, -OC_1-8Alkyl substituted;

R₁，R₂and R₃Each independently selected from H, halogen, CN, C_1-8Alkyl radical, -C_1-8Haloalkyl, or-OC_1-8Alkyl or

R₁And X and the atoms to which they are attached form a 5-to 6-membered heterocyclic ring; the heterocycle optionally comprises 1, 2 or 3 heteroatoms independently selected from N, S, or O; said heterocycle being optionally substituted by C_1-8Alkyl radical, -C_0-4alkyl-COOH, or-C_0-4alkyl-OH; or

R₃And Y and the atoms to which they are attached form a 5-to 6-membered heterocyclic ring; the heterocycle optionally comprises 1, 2 or 3 heteroatoms independently selected from N, S, or O; said heterocycle being optionally substituted by C_1-8Alkyl radical, -C_0-4alkyl-COOH, or-C_0-4alkyl-OH;

R₄and R₅Each independently selected from H, -CONH₂，-C_1-8Alkyl radical, -C_1-8Alkenyl, -C_1-8Haloalkyl, -C_1-8Heteroalkyl group, C_3-10Cycloalkyl, -C_1-4alkyl-C_5-6Aryl, -CO-C_1-4Alkyl, -SO₂-C_1-4Alkyl radical, -C_1-4alkyl-COOH, -C_1-4alkyl-OH; or

R₄And R₅And the atoms to which they are attached form a 5-to 6-membered heterocyclic ring; the heterocycle optionally comprises 1, 2 or 3 heteroatoms independently selected from N, S, or O; said heterocycle being optionally substituted by C_1-8Alkyl radical, -C_0-4alkyl-COOH, or-C_0-4alkyl-OH;

R₆and R₇Each independently selected from H, -C_1-8Alkyl radical, -C_1-8Heteroalkyl, or C_3-10Cycloalkyl radicals in which-C_1-8Alkyl radical, -C_1-8Heteroalkyl, or C_3-10Cycloalkyl is optionally substituted with-COOH or-OH; or

R₆And R₇And the atoms to which they are attached form a 4-to 6-membered heterocyclic ring; the heterocycle optionally comprises 1, 2 or 3 heteroatoms independently selected from N, S, or O; said heterocyclic ring being optionally substituted by-C_0-4alkyl-COOH, or-C_0-4alkyl-OH;

R₈，R₉，R₁₀each independently selected from H, -C_1-4Alkyl radical, -C_1-4Haloalkyl, -C_1-8Heteroalkyl group, -C_1-4alkyl-COOH, or-C_1-4alkyl-OH;

R₁₁and R₂₂Each independently selected from H, halogen, CN, or C_1-8An alkyl group; or

R₁₁And R₂₂And the atoms to which they are attached form a 3-to 4-membered carbocyclic ring;

n is 1, 2 or 3.

In some embodiments of formula i, ring a is a 6-membered heterocyclic ring containing 1, 2, or 3 heteroatoms independently selected from N, or S;

in some embodiments of formula i, ring B is a 6-membered heterocyclic ring containing 1, 2, or 3 heteroatoms independently selected from N, or S;

in some embodiments of the process of formula i,is that

In some embodiments of formula I, R₁And R₂Each independently selected from H, F, Cl, CN, or methyl.

In some embodiments of formula I, R₃Is H, F, Cl, CH₃Or CF₃。

In some embodiments of formula I, R₄And R₅And the atoms to which they are attached form a 5-to 6-membered heterocyclic ring. Preferably, the 5-to 6-membered heterocyclic ring isWherein the 5-to 6-membered heterocyclic ring may optionally be substituted by-COOH or-CH₃And (4) substituting.

In other embodiments of formula I, R₄And R₅Each independently selected from H, C_1-4Alkyl, -CO-C_1-4Alkyl, -SO₂-C_1-4Alkyl radical, -C_1-4alkyl-COOH, -C_1-4alkyl-OH.

In-situ typeIn some embodiments of I, R₆And R₇And the atoms to which they are attached form a 6-membered heterocyclic ring. Preferably, the heterocycle may be optionally substituted with-COOH.

In other embodiments of formula I, R₆And R₇Each independently selected from H, -C_1-8Alkyl radical, -C_1-8Heteroalkyl, or C_3-10Cycloalkyl radicals in which-C_1-8Alkyl radical, -C_1-8Heterocyclyl group, or C_3-10Cycloalkyl is optionally substituted with-COOH or-OH.

In some embodiments of formula I, R₈，R₉，R₁₀Each independently selected from H or methyl.

In some embodiments of formula I, R₁₁And R₂₂Each independently selected from H or methyl.

In other embodiments of formula I, R₁₁And R₂₂And the atoms to which they are attached make up a 3-to 4-membered carbocyclic ring.

In some embodiments of formula i, n is 1.

The present invention further provides preferred embodiments with respect to the compound of formula i, wherein the compound is:

1) ((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) glycine;

2) ((8- ((2-methyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) glycine;

3) ((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) proline;

4) ((8- ((2-methyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) proline;

5)1- ((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-2-yl) methyl) piperidine-2-acetic acid;

6)1- ((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) -4-hydroxypyrrolidine-2-acetic acid;

7)3- (((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) amino) propanoic acid;

8)4- (((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) amino) butanoic acid;

9)1- ((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) pyrrolidine-3-acetic acid;

10) ((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) serine;

11) n- ((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) -N-methylglycine;

12)1- ((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) -4, 4-difluoropyrrolidine-2-acetic acid;

13)2- (((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) amino) -3-hydroxy-2-methylpropionic acid;

14) n- ((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) -N-ethylglycine;

15)1- ((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) azetidine-3-acetic acid;

16)1- ((8- ((2,2' -dimethyl-3 ' - (3- (piperidin-1-yl) propoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

17)1- ((8- ((2,2' -dimethyl-3 ' - (3- (4-methylpiperazin-1-yl) propoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

18)1- ((8- ((2,2' -dimethyl-3 ' - (3- (pyrrolidin-1-yl) propoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

19)1- ((8- ((3' - (3- (3-hydroxypyrrolidin-1-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

20)1- ((8- ((2-methyl-3- (1- (3-morpholinopropyl) indolin-4-yl) phenyl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

21)1- ((8- ((3' - (3- (diethylamino) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

22)1- ((8- ((3' - (3- (benzyl (methyl) amino) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

23)1- ((8- ((3' - (3- (ethylamino) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

24)1- ((8- ((3' - (3-acetylaminopropoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

25)1- ((8- ((2,2' -dimethyl-3 ' - (3-ureidopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

26)1- ((8- ((3' - (3-guanidinopropoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

27)1- ((8- ((2,2' -dimethyl-3 ' - (3- (methylsulfonylamino) propoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

28)1- ((8- ((3' - (3- ((carboxymethyl) amino) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

29)1- ((8- ((3' - (3- (2-carboxypyrrolidin-1-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

30)1- ((8- ((3' - (3- (4-carboxypiperidin-1-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

31)1- ((8- ((2,2' -dimethyl-3 ' - (2-morpholinoethoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

32)1- ((8- ((2-methyl-3- (7- (2-morpholinoethoxy) naphthalen-2-yl) phenyl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

33)1- ((8- ((2-methyl-3 '- (3- (oxetan-3-ylamino) propoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

34)1- ((8- ((2-methyl-3- (1- (3-morpholinopropyl) -1H-indol-4-yl) phenyl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

35)1- ((8- ((2-methyl-3- (2- (2-morpholinoethyl) -1H-indol-6-yl) phenyl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

36)1- ((8- ((2-methyl-3 '- (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

37)1- ((8- ((2' -fluoro-2-methyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

38)1- ((8- ((2' -chloro-2-methyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

39)1- ((8- ((2' -cyano-2-methyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

40)1- ((8- ((4' -fluoro-2-methyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

41)1- ((8- ((2,2',4' -trimethyl-3 '- (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

42)1- ((8- ((2,4' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

43)1- ((8- ((4' -chloro-2-methyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

44) ((8- ((4' -chloro-2-methyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) glycine;

45) ((8- ((2' -chloro-2-methyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) glycine;

46) ((8- ((2-methyl-3- (4-methyl-5- (3-morpholinopropoxy) pyridin-3-yl) phenyl) amino) -1, 7-naphthyridin-3-yl) methyl) glycine;

47)1- ((8- ((2,2' -dimethyl-3 ' - ((2-morpholinoethoxy) methyl) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

48)1- ((8- ((2,2' -dimethyl-3 ' - ((3-morpholinopropyl) amino) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

49)1- ((8- ((2,2' -dimethyl-3 ' - (3-morpholinepropionamide) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

50)1- ((8- ((2,2' -dimethyl-3 ' - (N-methyl-3-morpholinepropionamide) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

51)1- ((8- ((3' - (3- (2- ((2-hydroxyethyl) amino) ethyl) urea) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

52)1- ((8- ((2,2' -dimethyl-3 ' - (methyl (3-morpholinopropyl) amino) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

53)1- ((8- ((2,2' -dimethyl-3 ' - (3- (2-morpholinoethyl) urea) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

54)1- ((8- ((2,2' -dimethyl-3 ' - ((1- (morpholinomethyl) cyclopropyl) methoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

55)1- ((8- ((2,2' -dimethyl-3 ' - ((4-morpholinebutan-2-yl) oxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

56)1- ((8- ((2-methyl-3- (1- (2-morpholinoethoxy) -2, 3-dihydro-1H-indan-4-yl) phenyl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

57) (E) -1- ((8- ((2,2' -dimethyl-3 ' - (4-morpholinobutyl-1-en-1-yl) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

58)1- ((8- ((2,2' -dimethyl-3 ' - (4-morpholinoethyl) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

59)1- ((8- ((4' -methoxy-2-methyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid;

60)1- ((8- ((2-methyl-3 ' - (3-morpholinopropoxy) -4' - (trifluoromethyl) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid.

The invention also provides a pharmaceutical composition comprising a compound of any of the present invention and a pharmaceutically acceptable carrier, such as hydroxypropylmethylcellulose. In the composition, the weight ratio of the compound to the adjuvant is about 0.0001 to 10.

The invention further provides the use of a pharmaceutical composition comprising formula I in the manufacture of a medicament for treating a disease in a subject.

The invention also provides some preferable technical schemes about the application.

In some embodiments, the prepared medicament is useful for treating, preventing or delaying the onset or progression of cancer, cancer metastasis, an immune system related disease. The cancer includes colon cancer, stomach cancer, thyroid cancer, lung cancer, leukemia, pancreatic cancer, melanoma, multiple melanoma, brain cancer, kidney cancer, prostate cancer, ovarian cancer or breast cancer.

The present invention provides methods of inhibiting the PD-1/PD-L1 interaction, comprising administering to a patient a compound of the present invention, or a pharmaceutically acceptable salt or stereoisomer thereof.

The present invention provides a method of treating a disease associated with inhibition of the PD-1/PD-L1 interaction, comprising administering to a patient in need thereof a therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable salt, stereoisomer. Wherein the disease is colon cancer, stomach cancer, thyroid cancer, lung cancer, leukemia, pancreatic cancer, melanoma, multiple melanoma, brain cancer, kidney cancer, prostate cancer, ovarian cancer or breast cancer.

The present invention provides a method of enhancing, stimulating and/or increasing an immune response in a patient, comprising administering to a patient in need thereof a therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable salt or stereoisomer thereof.

The invention also provides the use of a compound of the invention or a pharmaceutical composition thereof in the manufacture of a medicament.

In some embodiments, the medicament is for treating or preventing cancer.

In some embodiments, the cancer is colon cancer, gastric cancer, thyroid cancer, lung cancer, leukemia, pancreatic cancer, melanoma, multiple melanoma, brain cancer, renal cancer, prostate cancer, ovarian cancer, or breast cancer.

In some embodiments, the medicament is useful as a PD-1/PD-L1 interaction inhibitor.

The general chemical terms used in the above formula have their usual meanings. For example, the term "halogen" as used herein refers to fluorine, chlorine, bromine or iodine unless otherwise indicated. Preferred halogen groups include F, Cl and Br.

The term "alkyl" as used herein, unless otherwise specified, includes saturated monovalent hydrocarbon radicals having straight, branched or cyclic moieties. For example, alkyl includes methyl, ethyl, propyl, isopropyl, cyclopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, cyclobutyl, n-pentyl, 3- (2-methyl) butyl, 2-pentyl, 2-methylbutyl, neopentyl, cyclopentyl, n-hexyl, 2-methylpentyl and cyclohexyl. Similarly, as in C_1-8In alkyl, definition C_1-8To identify groups having 1, 2,3, 4,5, 6, 7 or 8 carbon atoms in a linear or branched arrangement.

Alkenyl and alkynyl groups include straight, branched or cyclic alkenes and alkynes. Likewise, "C_2-8Alkenyl "and" C_2-8Alkynyl "refers to a linear or branched arrangement of alkenyl or alkynyl groups having 2,3, 4,5, 6, 7 or 8 carbon atoms.

Alkoxy is an oxygen ether formed from the aforementioned linear, branched or cyclic alkyl groups.

The term "aryl" as used herein, unless otherwise specified, refers to an unsubstituted or substituted monocyclic or polycyclic ring system containing carbon ring atoms. Preferred aryl groups are monocyclic or bicyclic 6-10 membered aromatic ring systems. Phenyl and naphthyl are preferred aryl groups. The most preferred aryl group is phenyl.

As used herein, unless otherwise specified, the term "heterocyclyl" means an unsubstituted or substituted stable three to eight membered monocyclic saturated ring system consisting of carbon atoms and 1-3 heteroatoms selected from N, O or S, and wherein the nitrogen or sulfur heteroatoms may optionally be oxidized, and the nitrogen heteroatoms may optionally be quaternized. The heterocyclic group may be attached at any heteroatom or carbon atom which results in the formation of a stable structure. Examples of such heterocyclyl groups include, but are not limited to, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, oxopiperazinyl, oxopiperidinyl, oxaheptyl, heptyl, tetrahydrofuranyl, dioxolanyl, tetrahydroimidazolyl, tetrahydrothiazolyl, tetrahydrooxazolyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl sulfoxide, thiomorpholinyl sulfone, and oxadiazolyl.

As used herein, unless otherwise indicated, the term "heteroaryl" means an unsubstituted or substituted stable five or six membered monocyclic aromatic ring system or an unsubstituted or substituted nine or ten membered benzo-fused heteroaromatic ring system or a bicyclic heteroaromatic ring system, consisting of carbon atoms and 1 to 4 heteroatoms selected from N, O or S, wherein the nitrogen or sulfur heteroatoms may be optionally oxidized and the heteroatoms may be optionally quaternized. The heteroaryl group may be attached at any heteroatom or carbon atom, thereby forming a stable structure. Examples of heteroaryl groups include, but are not limited to, thienyl, furyl, imidazolyl, isoxazolyl, oxazolyl, pyrazolyl, pyrrolyl, thiazolyl, thiadiazolyl, triazolyl, pyridyl, pyridazinyl, indolyl, azaindolyl, indazolyl, benzimidazolyl, benzofuranyl, benzothienyl, benzisoxazolyl, benzoxazolyl, benzopyrazolyl, benzothiazolyl, benzothiadiazolyl, benzotriazolyl quinolinylbenzylidene.

The term "alkenyloxy" refers to the group-O-alkenyl, wherein alkenyl is as defined above.

The term "alkoxy" refers to the group-O-alkyl, wherein alkenyl is as defined above.

The term "cycloalkyl" refers to a cyclic saturated alkyl chain having 3 to 12 carbon atoms, such as cyclopropyl, cyclobutyl.

The term "substituted" refers to the substitution of one or more H atoms in a group with the same or different substituents, respectively. Common substituents include, but are not limited to, halogen (F, Cl, Br or I), C_1-8Alkyl radical, C_3-12Cycloalkyl, -OR¹，SR¹，＝O，＝S，-C(O)R¹，-C(S)R¹，＝NR¹，-C(O)OR¹，-C(S)OR¹，-NR¹R²，-C(O)NR¹R²Cyano, nitro, -S (O)₂R¹，-OS(O₂)OR¹，-OS(O)₂R¹，-OP(O)(OR¹)(OR²) (ii) a Wherein R is¹And R²Each independently selected from-H, lower alkyl, lower haloalkyl. In some embodiments, the substituents can be independently selected from-F, -Cl, -Br, -I, -OH, trifluoromethoxy, ethoxy, propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, -SCH₃，-SC₂H₅Formaldehyde group, -C (OCH)₃) Cyano, nitro, CF₃-OCF₃Amino, dimethylamino, methylthio, sulfonyl and acetyl.

The term "composition" as used herein refers to a product comprising the specified ingredients at the specified levels, as well as products which consist, directly or indirectly, of combinations of the specified ingredients at the specified levels. Accordingly, pharmaceutical compositions containing the compounds of the present invention as active ingredients, as well as methods for preparing the compounds of the present invention, are within the scope of the present invention. Additionally, the invention also includes crystalline forms of the compound that may exist in polymorphic forms. Also, some compounds may form solvates with water (hydrates) or common organic solvents, and such solvates are also within the scope of the present invention.

Examples of substituted alkyl groups include, but are not limited to, 2-aminoethyl, 2-hydroxyethyl, pentachloroethyl, trifluoromethyl, methoxymethyl, pentafluoroethyl, and piperazinylmethyl.

Examples of substituted alkoxy groups include, but are not limited to, substituted aminomethoxy, trifluoromethoxy, 2-xylenylaminoethoxy, 2-ethoxycarbonylethoxy, 3-hydroxypropoxy.

The compounds of the invention may also be present in the form of pharmaceutically acceptable salts. In pharmaceutical use, the salts of the compounds of the present invention refer to non-toxic pharmaceutically acceptable salts. Pharmaceutically acceptable salts include pharmaceutically acceptable acidic/anionic salts and basic/cationic salts. Pharmaceutically acceptable salts of acidic/anionic salts are generally employed in the form of basic nitrogen protonated by inorganic or organic acids. Representative organic or inorganic acids include hydrochloric, hydrobromic, hydrofluoric, perchloric, sulfuric, nitric, phosphoric, acetic, propionic, glycolic, lactic, succinic, maleic, fumaric, malic, tartaric, citric, benzoic, mandelic, methanesulfonic, hydroxyethanesulfonic, benzenesulfonic, oxalic, pamoic, 2-naphthalenesulfonic, p-toluenesulfonic, cyclamic, salicylic, saccharin or trifluoroacetic acid. Pharmaceutically acceptable basic/cationic salts include, but are not limited to, aluminum, calcium, chloroprocaine, choline, diethanolamine, ethylenediamine, lithium, magnesium, potassium, sodium, and zinc.

The present invention includes within its scope prodrugs of the compounds of the present invention. In general, such prodrugs are functional derivatives of the compounds that are readily converted in vivo to the desired compounds. Thus, in the methods of treatment of the present invention, the term "administering" shall include the use of a specifically disclosed compound or a compound that may not be specifically disclosed but which is converted to a particular compound to treat the various disorders described. Conventional methods for selecting and preparing suitable prodrug derivatives are described, for example, in "Design of Prodrugs", ed. Vol.5, pp.2, pp.3. Bundgaard, eiswei er, 1985.

The definition of any substituent or variable at a particular position in a molecule is independent of the definitions elsewhere in that molecule. It is to be understood that substituents and substitution patterns for the compounds of the present invention may be selected by one of ordinary skill in the art to provide compounds that are chemically stable and that can be readily synthesized by techniques and those methods known in the art.

The compounds encompassed by the present invention may contain one or more asymmetric centers and thus may give rise to diastereomers and optical isomers. The present invention includes all such possible diastereomers and racemic mixtures thereof, as well as substantially pure resolved enantiomers thereof, all possible geometric isomers thereof, and pharmaceutically acceptable salts thereof.

The compounds of formula I above do not contain a defined stereochemistry at a particular position. The present invention includes all stereoisomers of the compounds of formula i and pharmaceutically acceptable salts thereof. Also, mixtures of stereoisomers and isolated specific isomers thereof are included. The products of such procedures may be mixtures of stereoisomers during the synthetic procedures used to prepare such compounds, or during the use of racemic or epimeric procedures known to those skilled in the art.

When tautomers of compounds of formula i exist, the present invention includes any possible tautomers and pharmaceutically acceptable salts thereof and mixtures thereof, unless specifically stated otherwise.

When the compounds of formula i and pharmaceutically acceptable salts thereof are present in solvate or polymorphic form, the present invention includes any possible solvate and polymorphic form. The type of the solvent forming the solvate is not particularly limited as long as the solvent is pharmacologically acceptable. For example, water, ethanol, propanol, acetone, and the like can be used.

The term "pharmaceutically acceptable salts" refers to salts prepared from pharmaceutically acceptable non-toxic bases or acids. When the compounds of the present invention are acidic, their corresponding salts may be conveniently prepared from pharmaceutically acceptable non-toxic bases including inorganic and organic bases. Salts derived from such inorganic bases include aluminum, ammonium, calcium, copper (mono-and divalent), iron, ferrous, lithium, magnesium, manganese (mono-and divalent), potassium, sodium, zinc and the like salts. Particularly preferred are the ammonium, calcium, magnesium, potassium and sodium salts. Salts derived from pharmaceutically acceptable organic non-toxic bases include salts of primary, secondary and tertiary amines, as well as cyclic and substituted amines, such as naturally occurring and synthetic substituted amines. Other pharmaceutically acceptable organic non-toxic bases from which salts can be formed include ion exchange resins such as arginine, betaine, caffeine, choline, N ', N' -dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrazinoaniline, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, propylamine and the like.

When the compounds of the present invention are basic, their corresponding salts may be conveniently prepared from pharmaceutically acceptable non-toxic acids, including inorganic and organic acids. Such acids include, for example, acetic, benzenesulfonic, benzoic, camphorsulfonic, citric, ethanesulfonic, formic, fumaric, gluconic, glutamic, hydrobromic, hydrochloric, plasma, lactic, maleic, malic, mandelic, methanesulfonic, mucic, nitric, pamoic, pantothenic, phosphoric, succinic, sulfuric, tartaric, p-toluenesulfonic acid and the like. Preference is given to citric acid, hydrobromic acid, formic acid, hydrochloric acid, maleic acid, phosphoric acid, sulfuric acid and tartaric acid, particular preference to formic acid and hydrochloric acid. Since the compounds of formula I are intended for pharmaceutical use, they are preferably provided in substantially pure form, e.g. at least 60% pure, more suitably at least 75% pure, especially at least 98% pure (% by weight) basis).

The pharmaceutical composition of the present invention comprises a compound represented by formula I (or a pharmaceutically acceptable salt thereof) as an active ingredient, a pharmaceutically acceptable carrier and other optional therapeutic ingredients or adjuvants. The compositions include those suitable for oral, rectal, topical and parenteral (including subcutaneous, intramuscular and intravenous) administration, although the most suitable route of administration of the active ingredient in any given case will depend on the particular host, the nature and severity of the disease. The pharmaceutical compositions may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy.

In practice, the compounds represented by formula I of the present invention, or prodrugs thereof, or metabolites or pharmaceutically acceptable salts thereof, can be intimately admixed as the active ingredient with a pharmaceutical carrier according to conventional pharmaceutical compounding techniques. The carrier may take a wide variety of forms depending on the form of administration desired, for example, oral or parenteral (including intravenous). Thus, the pharmaceutical compositions of the present invention may be presented for oral administration in discrete dosage units, such as capsules, cachets, or tablets containing the specified levels of the active ingredient. In addition, the composition may be in the form of a powder, granules, a solution, a suspension in an aqueous solution, or in the form of an oil-in-water or water-in-oil in a non-aqueous solution. Furthermore, in addition to the various dosage forms described above, the compounds of formula I or pharmaceutically acceptable salts thereof may also be administered in a controlled release manner or by a delivery device. The composition can be prepared by any method of pharmacy. Generally, these methods include the step of bringing into association the active ingredient with one or more pharmaceutical carriers comprised of the desired ingredient. Generally, the compositions are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers. And the product is easily processed into a desired form.

Thus, the pharmaceutical compositions of the present invention may comprise a pharmaceutically acceptable carrier and a compound of formula I or a pharmaceutically acceptable salt thereof. The compounds of formula I or pharmaceutically acceptable salts thereof may also be included in combination with one or more other therapeutically effective active ingredients.

The pharmaceutically acceptable carrier may be solid, liquid or gaseous as desired. Solid carriers include lactose, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate and stearic acid. Liquid carriers include syrup, peanut oil, olive oil and water. The gaseous carrier includes carbon dioxide and nitrogen. In preparing oral dosage forms of the composition, any of the usual pharmaceutical media may be employed. For example, water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents, and the like can be used to form oral liquid preparations such as suspensions, tinctures, and solutions; carriers such as starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents and the like may be used to form oral solid preparations such as powders, capsules and tablets. Tablets and capsules using solid pharmaceutical carriers are the preferred oral dosage units. Optionally, the tablets may be coated by standard aqueous or non-aqueous techniques.

Tablets containing the composition may be prepared by compression or molding, optionally with one or more ingredients or adjuvants. Compression of the tablets may be carried out in suitable moulds and the active ingredient is optionally mixed with binders, lubricants, diluents, surfactants or other dispersing agents to form powders or granules with good flow properties. Molded tablets are prepared in a suitable machine and the mixture of powders may be moistened with an inert liquid wetting agent. Each tablet contains about 0.05mg to 5g of active ingredient, wherein a suitable and convenient carrier is present in an amount of about 5% to 95% by weight. A unit dose of the medicament contains approximately 1mg to 2mg of the active ingredient, typically 25mg,50mg, l00mg,200mg,300mg,400mg,500mg,600mg,800mg, or l000 mg.

Pharmaceutical compositions of the invention suitable for parenteral administration may be prepared as solutions or suspensions of the active compound in water. Suitable surfactants, such as hydroxypropyl cellulose, may be included. Dispersants may also be prepared in glycerol, liquid polyethylene glycols and mixtures thereof in oils. In addition, preservatives may also be included to prevent the unwanted growth of microorganisms.

Suitable compounds of the invention for administration by injection include sterile aqueous solutions and dispersions. Also, the compositions may be presented in sterile powder form for constitution with a sterile injectable solution or suspension. In all cases, the final injection form must be sterile and a liquid that is easy to inject. The pharmaceutical compositions must remain stable under the conditions of manufacture and storage. Thus, preservation environments are well established to prevent contamination by microorganisms such as bacteria and fungi. Solvents or dispersants which may serve as carriers may be water, ethanol, polyols (e.g. glycerol, propylene glycol and liquid polyethylene glycols), vegetable oils and suitable mixtures thereof.

The compounds of the present invention suitable for topical administration may generally be in the form of aerosols, creams, ointments, lotions, dusting powders and the like. And the composition is also suitable for transdermal mechanical administration. These formulations comprising a compound of formula I or a pharmaceutically acceptable salt thereof can be prepared by a convenient method. For example, a cream or ointment of the desired consistency can be prepared by mixing the hydrophilic material, water and about 5% to 10% of the compound.

The carrier of the compositions of the invention is solid when the composition is in the form for rectal administration. Preferably, the mixture is prepared as a unit dose suppository. Suitable carriers include cocoa butter or other carriers commonly used in the art. Suppositories may conveniently be formed by first mixing the composition with the softened or molten carrier, followed by cooling and shaping in a mould.

In addition to the aforementioned carriers, the aforementioned pharmaceutical formulations may also include one or more other carrier ingredients such as diluents, buffers, glidants, binders, surfactants, thickeners, lubricants, preservatives (including antioxidants), and the like. In addition, other adjuvants that can render the drug isotonic with the blood of the intended recipient are included. Compositions containing a compound of formula I or a pharmaceutically-acceptable salt thereof may also be prepared in the form of a powder or a liquid concentrate.

In general, in the above cases, it is usually a dosage level of from 0.01mg/kg to 150mg/kg per day, or from 0.5mg to 7g per patient per day. For example, colon cancer, rectal cancer, mantle cell lymphoma, multiple myeloma, breast cancer, prostate cancer, glioblastoma, squamous cell esophageal cancer, liposarcoma, T-cell lymphoma melanoma, pancreatic cancer, glioblastoma or lung cancer may be effectively treated by about 0.01 to 50mg of compound per kilogram of body weight per day, or about 0.5mg to about 3.5g of compound per patient per day.

It is understood, however, that the lowest or highest dose may be outside the ranges set forth above. The specific dose and treatment regimen for any one patient will depend upon a variety of factors including the activity of the specific compound, the age, body weight, general health, sex, diet, time of administration, route of administration, clearance rate, drug combination, the severity and course of the particular disease undergoing therapy, the mode of treatment for the patient, and the treatment regimen of the physician.

These and other aspects will become apparent from the following written description of the invention.

The following examples are provided to better illustrate the present invention. Unless otherwise expressly indicated, all parts and percentages are by weight and all temperatures are in degrees Celsius.

More specific details of the invention will be described by way of specific embodiments. The following examples are provided for illustrative purposes and are not intended to limit the invention in any way. Those skilled in the art will readily recognize a variety of non-critical parameters that may be altered or modified to produce substantially the same result. The compounds of the examples were found to inhibit the activity of PD-1/PD-L1 protein/protein interaction according to at least one of the assays described herein.

Detailed Description

The experimental procedures for preparing the compounds of the invention are described below. Some of the prepared compounds were purified on a Waters mass-directed fractionation system using open access preparative LCMS. The basic device settings, protocols and control software for the operation of these systems have been described in detail in the literature. See, e.g., Blom, "two pumps for preparative LC-MS in column dilution configuration", k.blom, j.combi.chem, 2002, 4, 295-; blom et al, "preparative LC-MS configuration and methods optimized for parallel synthetic purification", j.combi.chem, 2003, 5, 670-83; and Blom et al, "preparative LC-MS purification: improved compound-specific method optimization ", J.Combi.chem, 2004, 6, 874-883

The following abbreviations are used in the examples:

boc: t-tert-butoxycarbonyl;

BSA: bovine serum albumin;

DCM: dichloromethane;

DIEA: diisopropylethylamine;

DMF: n, N-dimethylformamide;

DMSO, DMSO: dimethyl sulfoxide;

Et₂o: diethyl ether;

EtOAc: ethyl acetate;

h or hrs: hours;

HATU: o- (7-azobenzotriazol) -N, N, N ', N' -tetramethyluronium hexafluorophosphate;

HTRF: homogeneous phase time-resolved fluorescence;

MeCN: methyl cyanide;

min: the method comprises the following steps of (1) taking minutes;

Pd(dppf)Cl.CH₂Cl₂:1, 1' -bis (diphenylphosphino) ferrocene-palladium (II) dichloride dichloromethane complex;

rt or r.t.: room temperature;

TFA: trifluoroacetic acid;

THF: tetrahydrofuran.

Scheme-general synthetic route to Compounds 1 and 2

EXAMPLE 1 Synthesis of Compound 1

((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) glycine

Step 1: preparation of 8-chloro-3-vinyl-1, 7-naphthyridine (M1)

To a solution of 3-bromo-8-chloro-1, 7-naphthyridine (2.43g) in toluene (30mL), EtOH (10mL), 10% Na₂CO₃Adding the solution (10mL)Pd(dppf)Cl₂DCM (420 mg). 4,4,5, 5-tetramethyl-2-vinyl-1, 3, 2-dioxaborane (3.1g) was added dropwise under nitrogen. The mixture was stirred at 100 ℃ for 16 hours. The reaction was quenched with water (50mL) and extracted 3 times with EtOAc. The organic phases were combined and washed with brine. The resulting solution was concentrated and purified by silica gel column (gradient from 8:1 to 5:1 with hexanes-EtOAc) to give 8-chloro-3-vinyl-1, 7-naphthyridine as a brown solid (1.1g) 88%).

Step 2: preparation of 8-chloro-1, 7-naphthyridine-3-carbaldehyde (M2)

To a solution of 1, 4-dioxane (20mL) and water (20mL) containing 8-chloro-3-vinyl-1, 7-naphthyridine (380mg) was added K₂OsO₄(4.0mg), followed by stirring at room temperature for 30 minutes. Adding a small amount of NaIO in portions at the same temperature₄(1.0 g). After stirring for 3 hours, the reaction was saturated with Na₂S₂O₃The solution was quenched. The mixture was extracted 3 times with DCM (40 mL). The organic phases were combined and washed with Na₂SO₄And (5) drying. The obtained solution is concentrated to obtain a crude product of 8-chloro-1, 7-naphthyridine-3-formaldehyde which can be directly used for the next reaction.

And step 3: preparation of methyl ((8-chloro-1, 7-naphthyridin-3-yl) methyl) glycinate (M3)

The above aldehyde compound was dissolved in DCM (20 mL). Glycine methyl ester hydrochloride (375mg) was added in one portion. After the resulting mixture was stirred at room temperature for 1 hour, STAB (420mg) was added in one portion at the same temperature. Stir at room temperature overnight. The resulting solution was saturated with Na₂C₂O₃The solution was quenched and extracted 3 times with DCM (20mL) and the organic phase with Na₂SO₄And (5) drying. The resulting solution was concentrated and purified by silica gel column (eluting with hexane-EtOAc in a gradient of 4:1 to 2: 1) to give ((methyl 8-chloro-1, 7-naphthyridin-3-yl) methyl)Glycine methyl ester (280mg) was a white solid.

And 4, step 4: preparation of methyl ((8- ((3-bromo-2-methylphenyl) amino) -1, 7-naphthyridin-3-yl) methyl) glycinate (M4)

To a microwave reaction flask were added 3-bromo-2-methylaniline (1.1g), ((8-chloro-1, 7-naphthyridin-3-yl) methyl) glycine methyl ester (0.9g) and t-BuOH (15.0 mL). The resulting mixture was stirred well at room temperature. A solution of 4M HCl in 1, 4-dioxane (3.0mL) was added dropwise. After the bottle cap is covered, the reaction is heated at 105 ℃ for 2 hours. Saturated Na₂C₂O₃The solution (30mL) was diluted and extracted with DCM (50mL × 2). The combined organic phases were washed with brine and then over MgSO₄Dried and concentrated in vacuo. The residue was purified by recrystallization from hexane: EtOAc ═ 8:1 to give ((8- ((3-bromo-2-methylphenyl) amino) -1, 7-naphthyridin-3-yl) methyl) glycine methyl ester (668mg) as a yellow solid.

And 5: preparation of 4- (3- (3-bromo-2-methylphenoxy) propyl) morpholine (M6)

To a solution of 3-bromo-2-methylphenyl (1.88g) in ACN (30mL) was added K₂CO₃(4.0 g). The mixture was stirred at room temperature for 30 minutes. 4- (3-chloropropyl) morpholine (3.0g) was added dropwise. The resulting solution was allowed to continue to react at room temperature overnight. After quenching the reaction with water, the mixture was extracted 3 times with EtOAc (100mL), the organic phases were combined and passed over Na₂SO₄And (5) drying. The resulting solution was concentrated and purified over silica gel column (hexane-EtOAc eluting with a gradient of 8:1 to 5: 1) to give 4- (3- (3-bromo-2-methylphenoxy) propyl) morpholine (2.5g) as a colorless oil.

Step 6: preparation of 4- (3- (2-methyl-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenoxy) propyl) morpholine (M7)

To a microwave reaction vial was added 4- (3- (3-bromo-2-methylphenoxy) propyl) morpholine (0.93g), bis (pinacolato) diboron (1.54g), Pd (dppf) Cl₂DCM (120mg), KOAC (1.0g) and 1, 4-dioxane (2.0 mL). After the bottle cap is closed, the reaction mixture is heated at 100 ℃ for 2 hours. Diluted with 50mL of water and extracted 2 times with DCM (60mL × 2). The combined organic extracts were washed with brine and then over MgSO₄Dried and concentrated in vacuo. The residue was purified by column on silica gel eluting with hexane-EtOAc in a gradient from 10:1 to 2:1 to give 4- (3- (2-methyl-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborane-2-yl) phenoxy) propyl) morpholine (580mg) as a brown oil.

Preparation of step 7 methyl ((8- (((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) glycinate (M5)

To a solution of ((8- (3-bromo-2-methylphenylamino) 1, 7-naphthyridin-3-yl) methyl) glycine methyl ester (233mg), 4- (3- (2-methyl-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborane-2-yl) phenoxy) propyl) morpholine (400mg) in toluene (6mL) under nitrogen atmosphere was added EtOH (2mL), 10% Na₂C₂O₃Solution (2mL), Pd (dppf) Cl₂DCM (18 mg). The mixture was stirred at 100 ℃ overnight. The reaction was quenched with water (20mL) and extracted 3 times with DCM. The organic phases were combined and washed with brine. The resulting solution was concentrated and purified by silica gel column (eluting with hexane-EtOAc in a gradient of 2:1 to 1: 2) to give ((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl)]-3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) glycinate methyl ester (180mg) was a brown semisolid.

And 8: preparation of ((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) glycine (Compound 1)

To a solution of ((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) glycinate (180mg) in 1:1(20mL) of THF/water was added NaOH (40 mg). The resulting mixture was stirred at room temperature for 24 hours. The reaction was quenched with 2M HCI and then the pH was adjusted to 4-5. THF and water were removed by evaporation. The resulting solid was purified by RP column (mobile phase MeCN: water (0.1% HCl) in a gradient from 10:90 to 30:70) to give ((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) glycine (88mg) as a white solid.

EXAMPLE 2 Synthesis of Compound 2

((8- ((2-methyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) glycine

Step 1: preparation of methyl ((8- ((2-methyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) glycinate

This compound was prepared using a method analogous to that described for M5 in example 1, substituting phenylboronic acid for M7. The resulting compound was purified by prep-TLC (EtOC: n-hexane ═ 1:1) to give ((8- ((2-methyl- [1,1' -biphenyl ] -3-yl ] amino ] -1, 7-naphthyridin-3-yl) methyl) glycine methyl ester (150mg) as a yellow solid.

Step 2: ((8- ((2-methyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) glycine (Compound 3)

This compound was prepared using a similar method to that described for compound 1. The resulting compound was purified by RP column (mobile phase: MeCN: water (0.1% HCl) in a gradient of 40:60 to 50: 50) to give ((8- ((2-methyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) glycine as a white solid (98 mg).

Scheme 2 synthetic route to Compound 5

EXAMPLE 3 Synthesis of Compound 5

1- ((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid

Step 1: preparation of (8-chloro-1, 7-naphthyridin-3-yl) methanol (M11)

The above aldehyde (620mg) was dissolved in MeOH. Adding NaBH in one time₄(400 mg). The mixture was stirred at room temperature for 2 hours and then quenched with water (30 mL). The mixture was extracted 3 times with DCM (20mL) and the organic phase was passed over Na₂SO₄And (5) drying. The resulting solution was concentrated and purified by silica gel column (n-hexane-EtOAc eluting with a gradient of 2:1 to 1:1) to give (8-chloro-1, 7-naphthyridin-3-yl) methanol (500mg) as a brown solid.

Step 2: preparation of (8- ((3-bromo-2-methylphenyl) amino) -1, 7-naphthyridin-3-yl) methanol (M12)

To a microwave reaction vial was added 3-bromo-2-methylaniline (370mg), (8-chloro-1, 7-naphthyridin-3-yl) methylAlcohol (98mg), LiHMDS (1.0M in THF, 4.0mL) and THF (3.5 mL). After the bottle cap is covered, the mixture is heated to 60 ℃ to react for 4 hours. Diluted with 20mL of water and extracted with DCM (20mL × 2). The combined organic phases were washed with brine, over MgSO₄Dried and concentrated in vacuo. The residue was purified by RP column (mobile phase: MeCN: water ═ 30:70) to give (8- ((3-bromo-2-methylphenyl) amino) -1, 7-naphthyridin-3-yl) methanol (73mg) as a brown solid.

And step 3: preparation of (8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methanol (M13)

This compound was prepared using a method analogous to that described for M5 in example 1, with M12 replacing M4. The resulting mixture was purified by prep-TLC (EtOAc: n-hexane ═ 1:1) to give 8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methanol as a yellow solid.

And 4, step 4: preparation of methyl 1-8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetate (M14)

To a solution of 8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methanol (82mg), TEA (100mg) in DCM (5.0mL) was added MsCl (80mg) dropwise at 0 ℃. The reaction was stirred at room temperature for 90 minutes. The mixture was concentrated under vacuum and dissolved in THF (3 mL). After the addition of methylpiperidine-2-acetate (50mg) and KI (1mg), the reaction was stirred at room temperature for 2 hours until the methanesulfonate salt was used up. The residue was concentrated and purified by RP-column (mobile phase: MeCN: water ═ 10:90 and 0.1% HCl) to give methyl 1-8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetate (44mg) as an off-white solid.

And 5: preparation of 1- ((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid (Compound 7)

This compound was prepared using a similar method to that described for compound 1. The resulting mixture was purified by RP-column (mobile phase: MeCN: water (0.1% HCl) at 30:70 to 40: 60) to give 1- ((8- ((2,2' -dimethyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid as a white solid (20 mg).

Scheme 3 synthetic route to Compound 19

EXAMPLE 4 Synthesis of Compound 19

1- ((8- ((3' - (3- (3-hydroxypyrrolidin-1-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid

Step 1: preparation of 1-bromo-3- (3-bromopropoxy) -2-methylbenzene (M21)

To a solution of 3-bromo-2-methylphenol (1.88g) in DMF (30mL) was added K₂CO₃(4.0 g). The mixture was stirred at room temperature for 30 minutes. 1, 3-dibromopropane (5.0g) was added dropwise. The resulting solution was allowed to continue to react at room temperature overnight. The reaction was quenched with water and extracted 3 times with EtOAc (100 mL). The organic phases were combined and passed over Na₂SO₄And (5) drying. Concentration stationThe solution was obtained and purified using silica gel column (eluting with n-hexane-EtOAc in a gradient of 20:1 to 10: 1) to give 1-bromo-3- (3-bromopropoxy) -2-methylbenzene (2.1g, crude).

Step 2: preparation of 1- (3- (3-bromo-2-methylphenoxy) propylpyrrolidin-3-ol (M22)

To a solution of ACN (40mL) containing 1-bromo-3- (3-bromopropoxy) -2-methylbenzene (2.2g) was added K₂CO₃(4.0g) and pyrrolidin-3-ol (3.2 g). The resulting solution was allowed to continue to react overnight at 45 ℃. The reaction was quenched with water. The mixture was extracted 3 times with EtOAc (100 mL). The organic phases were combined and passed over Na₂SO₄And (5) drying. The resulting solution was concentrated and purified by silica gel column (eluting with n-hexane-EtOAc in a 4:1 gradient) to give 1- (3- (3-bromo-2-methylphenoxy) propyl) pyrrolidin-3-ol (1.8g) as a colorless oil.

And step 3: preparation of 1- (3- (2-methyl-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenoxy) -propyl) pyrrolidin-3-ol (M23)

This compound was prepared using a method analogous to that described for M7 in example 1, substituting M22 for M6. The resulting mixture was purified by silica gel column eluting with n-hexane-EtOAc in a ratio of 2:1 to give 1- (3- (2-methyl-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborane-2-yl) phenoxy) -propyl) pyrrolidin-3-ol as a brown oil.

And 4, step 4: methyl 1- ((8- ((3' - (3- (3-hydroxypyrrolidin-1-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetate (M24)

This compound was prepared using a method analogous to that described for M5 in example 1, with M23 replacing M4. The resulting mixture was purified by prep-tlc (etoac) to give methyl 1- ((8- ((3' - (3- (3-hydroxypyrrolidin-1-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetate as a yellow semisolid.

And 5: 1- ((8- ((3' - (3- (3-hydroxypyrrolidin-1-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid (Compound 21)

This compound was prepared using a similar method to that described for compound 1. The resulting mixture was purified by RP column (mobile phase: MeCN: water (0.1% HCl) in a gradient of 30:70 to 40: 60) to give 1- ((8- ((3' - (3- (3-hydroxypyrrolidin-1-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) amino) -1, 7-naphthyridin-3-yl) methyl) piperidine-2-acetic acid (58 mg).

The following examples (shown in table 1) were prepared essentially as in examples 1, 2,3 or 4 using the corresponding starting materials.

TABLE 1

Homogeneous time-resolved fluorescence (HTRF) binding assays

The assay was performed in standard black 384 well polystyrene plates with a final volume of 20 μ L. Inhibitors were first serially diluted in DMSO and then added to the plate wells before the addition of the other reaction components. The final concentration of DMSO was determined to be 1%. The assay was performed at 25 ℃ in PBS buffer (pH 7.4) containing 0.05% Tween-20 and 0.1% BSA. Recombinant human PD-L1 protein (19-238) with a His-tag at the C-terminus was purchased from Acro biosystems (PD 1-H5229). Recombinant human PD-1 protein (25-167) with an Fc tag at the C-terminus was also purchased from Acrobios systems (PD 1-H5257). PD-L1 and PD-1 protein were diluted in assay buffer and 10. mu.L was added to the plate wells. The plates were centrifuged and the proteins were preincubated with inhibitors for 40 minutes. After incubation, 10. mu.L of HTRF detection buffer supplemented with an encrypted labeled anti-human IgG specific for Fc (Perkinelmer-AD0212) andallophycocyanin (allophycycanin) (APC, PerkinElmer-AD0059H) conjugated anti-His antibody. After centrifugation, the plates were incubated at 25 ℃ for 60 minutes. Before reading on a PHERAStar FS plate reader (665nm/620nm ratio). The final concentrations in the assay were-3 nM PD1, 10nM PD-L1, 1nM anti-human IgG and 20nM anti-His-allophycocyanin. IC was performed using GraphPad Prism 5.0 software to fit a curve controlling percent activity versus log inhibitor concentration₅₀And (4) measuring.

As exemplified in the examples, the IC of the compounds of the invention₅₀The values are in the following ranges: "+" stands for "IC₅₀Less than or equal to 2 nM; ". indicates" 2nM<IC₅₀Less than or equal to 10 nM; ". indicates" 10nM<IC₅₀Less than or equal to 150 nM; ". indicates" IC₅₀＞150nM”.

Table 1 provides data for the compound described in example A as determined using the PD-1/PD-L1 Homogeneous Time Resolved Fluorescence (HTRF) binding method.

For convenience of explanation, the general structure is shown below. Surprisingly, we found that "R" has a large effect on biological activity. As shown in table 1, we can see from the above example compounds (R in the example compounds includes a morpholine group), for example, compound 1, compound 3 or compound 5 have better activity.