Biomarkers associated with immune checkpoint inhibitor therapy and methods of use thereof

Abstract

The present disclosure relates to biomarkers that predict responsiveness of a patient's cancer to immune checkpoint inhibitor therapy. In some embodiments, the present disclosure provides diagnostic and / or prognostic methods using such biomarkers for determining whether a cancer patient will benefit from treatment with an aryl hydrocarbon receptor antagonist. In some embodiments, the biomarkers described herein can be used to inform and provide effective treatment methods for treating cancer, which include administration of an aryl hydrocarbon receptor antagonist, optionally in combination with immune checkpoint inhibitor therapy.

Description

[0001] This application claims priority to U.S. Provisional Patent Application No. 63 / 091,192, filed October 13, 2020, and U.S. Provisional Patent Application No. 63 / 107,309, filed October 29, 2020, the contents of which are incorporated herein by reference in their entirety.

[0002] This disclosure relates to biomarkers for predicting a patient’s cancer response to immune checkpoint inhibitor (ICI) therapy. In some embodiments, this disclosure provides diagnostic and / or prognostic methods using such biomarkers, comprising (a) measuring, or having measured, the amount of at least one biomarker from a serum, plasma, or stool sample obtained from a cancer patient; (b) comparing, or having compared, the amount of the at least one biomarker to a threshold fraction; and (c) determining that the cancer patient will benefit from treatment with an aryl hydrocarbon receptor (AhR) antagonist if the amount of the at least one biomarker is above the threshold fraction. A further aspect of this disclosure provides methods for treating cancer, comprising administering an AhR antagonist, optionally in combination with ICI therapy.

[0003] ICI therapy represents the first-line treatment standard for several cancers (Gandhi et al., N Engl J Med., 2018, 378(22), 2078-2092; Socinski et al., N Engl J Med., 2018, 378(24), 2288-2301; Antonia et al., N Engl J Med., 2017, 377(20), 1919-1929; Wolchok et al., N Engl J Med., 2017, 377(14), 1345-1356; Cella et al., Lancet Oncol., 2019, 20(2), 297-310). Treatments using this type of therapy aim to enhance the host's immune response to different stages of tumor progression, with fewer off-target effects compared to chemotherapy drugs (Tsai et al., Journal of Biomedical Science, 2017, 24(1), 35). Major ICI targets include programmed death 1 (PD-1), programmed death-ligand 1 (PD-L1), cytotoxic T-lymphocyte antigen 4 (CTLA-4), lymphocyte activation gene-3 (LAG-3), T-cell immunoglobulin and mucin-containing domain-3 (TIM-3), T-cell immunoglobulin and ITIM domain (TIGIT), and T-cell activation V-domain Ig inhibitor (VISTA).

[0004] Although ICI therapy has demonstrated significant clinical benefit for treating a variety of solid tumors, a lack of response and / or development of resistance have been observed in many patients (Horsman et al., Int J Mol Sci., 2020, 21(13), 4778). Furthermore, ICI therapy is often expensive and can cause inflammatory toxicity in these patients (Das et al., J Immunother Cancer., 2019, 7(1), 306). Therefore, it is important to determine the probability of response to ICI therapy in order to design treatment strategies that can overcome primary or secondary resistance to this type of therapy.

[0005] Microbial metabolites play a role in patient responses to ICI therapy. The gut microbiome is a collection of diverse microorganisms that typically reside in specific regions of the gastrointestinal tract (Dieterich et al., Med Sci [Medical Sciences] (Basel), 2018, 6(4):116; Donaldson et al., Nat Rev Microbiol. [Nature Reviews Microbiol], 2016, 14(1):20-32). In recent years, the relationship between the gut microbiome and overall host health has been a hot topic in medical research (Lin et al., BMC Immunol. [BMC Immunology], 2017, 18(1):2). In addition to playing a role in homeostasis within the intestinal barrier, the gut microbiome also plays a role in regulating immune function, with dysregulation of the gut microbiome establishing a correlation with certain inflammatory and malignant disease states of the gastrointestinal system. The gut microbiota regulates immune responses at different levels, influencing neutrophil migration, promoting the differentiation of T cells in Th1, Th2, and Th17 or regulatory T cells (Tregs), and modulating immune homeostasis and inflammation (Rossi et al., Int Rev Immunol. [International Review of Immunology], 2013, 32(5-6):471-474; Pandiyan et al., Front Immunol. [Fronts in Immunology], 2019, 10:426).

[0006] Response to ICI therapy is impaired due to alterations in the microbiome. Mice with solid tumors treated with sterile and antibiotic methods showed ineffective responses to ICI therapy, typically associated with reduced cellular immune responses and decreased cytokine production (Gopalakrishnan et al., Science, 2018, 359(6371):97-103). Administration of a probiotic blend of Bifidobacterium restored a more effective response to ICI therapy in a mouse cancer model (Matson et al., Science, 2018, 359(6371):104-108). Fecal metagenomic analysis in patients with melanoma revealed a distinct microbiome composition in responders to ICI therapy compared to non-responders, with responders exhibiting an abundance of *Bifidobacterium longum*, *Collinsella aerofaciens*, and *Enterococcus faecium* (Matson et al., Science, 2018, 359(6371):104-108). Similarly, microbiome composition analysis in humans (particularly enrichment of *Akkermansia muciniphila* and *Alistipes*) has been used to predict the ability to respond to anti-PD-1 / PD-L1 therapy in solid epithelial tumors such as non-small cell lung cancer (Routy et al., Science, 2018, 359(6371):91-97). In melanoma patients, high abundance of Clostridium, Ruminococcus, or Faecalibacterium species has been detected via CD4. + and CD8 + T cell activation and regulation of Treg and myeloid-derived suppressor cells are associated with a better response to PD-1 (Gopalakrishnan et al., Science, 2018, 359(6371):97-103). Transplantation of fecal microbiota from responder patients into germ-free (GF) mice improved ICI efficacy.

[0007] Therefore, preclinical studies in mice and early observational human cohorts have revealed the complex interactions between bacteria and the immune host response in antitumor activity. Identification of biomarkers, such as microbial metabolites, that predict the responsiveness of cancer patients to ICI therapy remains crucial. Such biomarkers could provide valuable information for determining the effectiveness of ICI therapy and for incorporating these findings into improved treatment strategies for cancer management.

[0008] This disclosure relates to biomarkers for predicting a patient's cancer's responsiveness to ICI therapy. In some embodiments, this disclosure provides methods for determining whether a cancer patient will benefit from treatment with an AhR antagonist. In some embodiments, this disclosure provides methods for predicting a patient's cancer's responsiveness to ICI therapy. Aspects of this disclosure further provide improved methods for treating cancer patients, including administering an AhR antagonist to the patient, optionally in combination with ICI therapy. In some embodiments, the patient has previously been treated with ICI therapy but has not responded to it. In some embodiments, the patient has previously been treated with ICI therapy and has developed resistance to it.

[0009] Certain embodiments of this disclosure are summarized in the following paragraphs. This list is exemplary only and does not exhaustively list all embodiments provided by this disclosure.

[0010] Example 1. A method for treating cancer in a patient in need, comprising: (a) measuring or having measured the amount of at least one aryl hydrocarbon receptor (AhR) ligand from a serum, plasma, or fecal sample obtained from the patient; (b) comparing or having compared the amount of the at least one AhR ligand to a threshold fraction; (c) determining or having determined that the patient will benefit from treatment with an AhR antagonist if the amount of the at least one AhR ligand is above the threshold fraction; and (d) treating the patient determined to benefit in (c) with an effective amount of the AhR antagonist.

[0011] Example 2. The method described in Example 1, wherein the patient suffers from non-small cell lung cancer, small cell lung cancer, head and neck squamous cell carcinoma, renal cell carcinoma, gastric adenocarcinoma, nasopharyngeal carcinoma, urothelial carcinoma, colorectal cancer, pleural mesothelioma, triple-negative breast cancer, esophageal tumor, multiple myeloma, gastric and gastroesophageal junction cancer, melanoma, Hodgkin lymphoma, hepatocellular carcinoma, lung cancer, head and neck cancer, non-Hodgkin lymphoma, metastatic clear cell renal cell carcinoma, squamous cell carcinoma of the lung, mesothelioma, gastric cancer, gastroesophageal junction cancer, metastatic melanoma, metastatic non-cutaneous melanoma, urinary... Epithelial carcinoma, diffuse large B-cell lymphoma, renal cell carcinoma, ovarian cancer, fallopian tube cancer, peritoneal tumors, extensive-stage small cell lung cancer, bladder cancer, transitional cell carcinoma, prostate cancer, recurrent or metastatic PD-L1 positive or negative squamous cell carcinoma of the head and neck, recurrent squamous cell lung cancer, advanced solid malignancies, pharyngeal squamous cell carcinoma, laryngeal squamous cell carcinoma, unresectable or metastatic melanoma, biliary tract tumors, esophageal squamous cell carcinoma, breast cancer, pancreatic cancer, glioblastoma, metastatic cancer, prostate cancer, solid organ cancer, gastric cancer, colon cancer, or liver cancer.

[0012] Example 3. The method as described in Example 1 or Example 2, wherein the patient has been treated with immune checkpoint inhibitor (ICI) therapy but has not responded to ICI therapy.

[0013] Example 4. The method as described in Example 1 or Example 2, wherein the patient has been treated with ICI therapy and has developed resistance to ICI therapy.

[0014] Example 5. The method as described in any one of Examples 1-4, wherein the at least one AhR ligand comprises kynurenine, tryptophan, phenylpyruvic acid, indolelacic acid, or a combination thereof.

[0015] Example 6. The method as described in Example 5, wherein the at least one AhR ligand comprises kynurenine, tryptophan, phenylpyruvic acid, and indolelacic acid.

[0016] Example 7. The method as described in any one of Examples 1-6, wherein the amount of the at least one AhR ligand in the sample is measured by liquid chromatography-mass spectrometry or gas chromatography-mass spectrometry.

[0017] Example 8. The method as described in any one of Examples 1-7, wherein the threshold fraction is determined by measuring the amount of at least one corresponding AhR ligand in a control serum, plasma, or fecal sample of a patient who has responded to ICI therapy.

[0018] Example 9. The method as described in any one of Examples 1-8, wherein the AhR antagonist is a compound of Formula I:

[0019] Or its pharmaceutically acceptable salt.

[0020] And among them:

[0021] R 1 and R 2 Each of these is independently selected from optionally substituted alkyl groups, optionally substituted ester groups, optionally substituted heteroalkyl groups, optionally substituted acyl groups, optionally substituted amide groups, optionally substituted aryl groups, optionally substituted heteroaryl groups, optionally substituted cycloalkyl groups, optionally substituted amine groups, and optionally substituted heterocycloalkyl groups.

[0022] R 3 The group is selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, optionally substituted amide, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted ester, optionally substituted heteroalkyl, optionally substituted heteroaryl, optionally substituted heterocycloalkyl, optionally substituted amine, cyano, halogen, hydroxyl and -C(O)H.

[0023] Example 10. The method as described in any one of Examples 1-9, wherein the AhR antagonist is a compound of formula Ia:

[0024] Or its pharmaceutically acceptable salt.

[0025] And among them:

[0026] Ring A is selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, and optionally substituted heterocyclic alkyl.

[0027] Ring B is selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, and optionally substituted heterocycloalkyl.

[0028] R is selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, optionally substituted amide, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted ester, optionally substituted heteroalkyl, optionally substituted heteroaryl, optionally substituted heterocycloalkyl, amino, cyano, halogen, hydroxyl and -C(O)H.

[0029] Example 11. The method as described in any one of Examples 1-9, wherein the AhR antagonist is any one of those listed in Table 2 or a pharmaceutically acceptable salt thereof.

[0030] Example 12. An in vitro method for determining whether a patient will benefit from treatment with an aryl hydrocarbon receptor (AhR) antagonist, comprising: (a) measuring or having measured the amount of at least one AhR ligand from a serum, plasma, or fecal sample obtained from the patient; (b) comparing or having compared the amount of the at least one AhR ligand with a threshold fraction; and (c) if the amount of the at least one AhR ligand is higher than the threshold fraction, determining that the patient will benefit from treatment with an AhR antagonist.

[0031] Example 13. The method as described in Example 12, wherein the patient has non-small cell lung cancer, small cell lung cancer, head and neck squamous cell carcinoma, renal cell carcinoma, gastric adenocarcinoma, nasopharyngeal carcinoma, urothelial carcinoma, colorectal cancer, pleural mesothelioma, triple-negative breast cancer, esophageal tumor, multiple myeloma, gastric and gastroesophageal junction cancer, melanoma, Hodgkin lymphoma, hepatocellular carcinoma, lung cancer, head and neck cancer, non-Hodgkin lymphoma, metastatic clear cell renal cell carcinoma, lung squamous cell carcinoma, mesothelioma, gastric cancer, gastroesophageal junction cancer, metastatic melanoma, metastatic non-cutaneous melanoma, Urothelial carcinoma, diffuse large B-cell lymphoma, renal cell carcinoma, ovarian cancer, fallopian tube cancer, peritoneal tumor, extensive-stage small cell lung cancer, bladder cancer, transitional cell carcinoma, prostate tumor, recurrent or metastatic PD-L1 positive or negative squamous cell carcinoma of the head and neck, recurrent squamous cell lung cancer, advanced solid malignant tumors, pharyngeal squamous cell carcinoma, laryngeal squamous cell carcinoma, unresectable or metastatic melanoma, biliary tract tumors, esophageal squamous cell carcinoma, breast cancer, pancreatic cancer, glioblastoma, metastatic cancer, prostate cancer, solid organ cancer, gastric cancer, colon cancer, or liver cancer.

[0032] Example 14. The method as described in Example 12 or Example 13, wherein the patient has been treated with immune checkpoint inhibitor (ICI) therapy but has not responded to ICI therapy.

[0033] Example 15. The method as described in Example 12 or Example 13, wherein the patient has been treated with ICI therapy and has developed resistance to ICI therapy.

[0034] Example 16. The method as described in any one of Examples 12-15, wherein the at least one AhR ligand comprises kynurenine, tryptophan, phenylpyruvic acid, indolelacic acid, or a combination thereof.

[0035] Example 17. The method as described in Example 16, wherein the at least one AhR ligand comprises kynurenine, tryptophan, phenylpyruvic acid, and indolelacic acid.

[0036] Example 18. The method as described in any one of Examples 12-17, wherein the amount of the at least one AhR ligand in the sample is measured by liquid chromatography-mass spectrometry or gas chromatography-mass spectrometry.

[0037] Example 19. The method as described in any one of Examples 12-18, wherein the threshold fraction is determined by measuring the amount of at least one corresponding AhR ligand in a control serum, plasma, or fecal sample of a patient who has responded to ICI therapy.

[0038] Example 20. The method as described in any one of Examples 11-19, wherein the AhR antagonist is a compound of Formula I:

[0039] Or its pharmaceutically acceptable salt.

[0040] And among them:

[0041] R 1 and R 2 Each of these is independently selected from optionally substituted alkyl groups, optionally substituted ester groups, optionally substituted heteroalkyl groups, optionally substituted acyl groups, optionally substituted amide groups, optionally substituted aryl groups, optionally substituted heteroaryl groups, optionally substituted cycloalkyl groups, optionally substituted amine groups, and optionally substituted heterocycloalkyl groups.

[0042] R 3 The group is selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, optionally substituted amide, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted ester, optionally substituted heteroalkyl, optionally substituted heteroaryl, optionally substituted heterocycloalkyl, optionally substituted amine, cyano, halogen, hydroxyl and -C(O)H.

[0043] Example 21. The method as described in any one of Examples 12-20, wherein the AhR antagonist is a compound of formula Ia:

[0044] Or its pharmaceutically acceptable salt.

[0045] And among them:

[0046] Ring A is selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, and optionally substituted heterocyclic alkyl.

[0047] Ring B is selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, and optionally substituted heterocycloalkyl.

[0048] R is selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, optionally substituted amide, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted ester, optionally substituted heteroalkyl, optionally substituted heteroaryl, optionally substituted heterocycloalkyl, amino, cyano, halogen, hydroxyl and -C(O)H.

[0049] Example 22. The method as described in any one of Examples 12-20, wherein the AhR antagonist is any one of those listed in Table 2 or a pharmaceutically acceptable salt thereof.

[0050] Example 23. A method of treating cancer in a patient in need with a combination therapy comprising an immune checkpoint inhibitor (ICI) and an aryl hydrocarbon receptor (AhR) antagonist, comprising: (a) measuring or having measured the amount of at least one AhR ligand from a serum, plasma, or fecal sample obtained from the patient; (b) comparing or having compared the amount of the at least one AhR ligand with a threshold fraction; (c) determining or having determined that the patient will benefit from the combination therapy if the amount of the at least one AhR ligand is above the threshold fraction; and (d) treating the patient determined to benefit in (c) with the combination therapy.

[0051] Example 24. The method described in Example 23, wherein the patient has non-small cell lung cancer, small cell lung cancer, head and neck squamous cell carcinoma, renal cell carcinoma, gastric adenocarcinoma, nasopharyngeal carcinoma, urothelial carcinoma, colorectal cancer, pleural mesothelioma, triple-negative breast cancer, esophageal tumor, multiple myeloma, gastric and gastroesophageal junction cancer, melanoma, Hodgkin lymphoma, hepatocellular carcinoma, lung cancer, head and neck cancer, non-Hodgkin lymphoma, metastatic clear cell renal cell carcinoma, lung squamous cell carcinoma, mesothelioma, gastric cancer, gastroesophageal junction cancer, metastatic melanoma, metastatic non-cutaneous melanoma, Urothelial carcinoma, diffuse large B-cell lymphoma, renal cell carcinoma, ovarian cancer, fallopian tube cancer, peritoneal tumor, extensive-stage small cell lung cancer, bladder cancer, transitional cell carcinoma, prostate tumor, recurrent or metastatic PD-L1 positive or negative squamous cell carcinoma of the head and neck, recurrent squamous cell lung cancer, advanced solid malignant tumors, pharyngeal squamous cell carcinoma, laryngeal squamous cell carcinoma, unresectable or metastatic melanoma, biliary tract tumors, esophageal squamous cell carcinoma, breast cancer, pancreatic cancer, glioblastoma, metastatic cancer, prostate cancer, solid organ cancer, gastric cancer, colon cancer, or liver cancer.

[0052] Example 25. The method as described in Example 23 or Example 24, wherein the patient has been treated with ICI therapy but is unresponsive to ICI therapy.

[0053] Example 26. The method as described in Example 23 or Example 24, wherein the patient has been treated with ICI therapy and has developed resistance to ICI therapy.

[0054] Example 27. The method of any one of Examples 23-26, wherein the at least one AhR ligand comprises kynurenine, tryptophan, phenylpyruvic acid, indolelacic acid, or a combination thereof.

[0055] Example 28. The method as described in Example 27, wherein the at least one AhR ligand comprises kynurenine, tryptophan, phenylpyruvic acid, and indolelacic acid.

[0056] Example 29. The method as described in any one of Examples 23-28, wherein the amount of the at least one AhR ligand in the sample is measured by liquid chromatography-mass spectrometry or gas chromatography-mass spectrometry.

[0057] Example 30. The method as described in any one of Examples 23-29, wherein the ICI therapy comprises an anti-PD-1 antibody, an anti-PD-L1 antibody, an anti-CTLA-4 antibody, an anti-LAG-3 antibody, an anti-TIM-3 antibody, an anti-TIGIT antibody, an anti-VISTA antibody, or a combination thereof.

[0058] Example 31. The method as described in any one of Examples 23-30, wherein the threshold fraction is determined by measuring the amount of at least one corresponding AhR ligand in a control serum, plasma, or fecal sample of a patient who has responded to ICI therapy.

[0059] Example 32. The method as described in any one of Examples 23-31, wherein the AhR antagonist is a compound of Formula I:

[0060] Or its pharmaceutically acceptable salt.

[0061] And among them:

[0062] R 1 and R 2 Each of these is independently selected from optionally substituted alkyl groups, optionally substituted ester groups, optionally substituted heteroalkyl groups, optionally substituted acyl groups, optionally substituted amide groups, optionally substituted aryl groups, optionally substituted heteroaryl groups, optionally substituted cycloalkyl groups, optionally substituted amine groups, and optionally substituted heterocycloalkyl groups.

[0063] R 3 The group is selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, optionally substituted amide, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted ester, optionally substituted heteroalkyl, optionally substituted heteroaryl, optionally substituted heterocycloalkyl, optionally substituted amine, cyano, halogen, hydroxyl and -C(O)H.

[0064] Example 33. The method as described in any one of Examples 23-32, wherein the AhR antagonist is a compound of formula Ia:

[0065] Or its pharmaceutically acceptable salt.

[0066] And among them:

[0067] Ring A is selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, and optionally substituted heterocyclic alkyl.

[0068] Ring B is selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, and optionally substituted heterocycloalkyl.

[0069] R is selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, optionally substituted amide, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted ester, optionally substituted heteroalkyl, optionally substituted heteroaryl, optionally substituted heterocycloalkyl, amino, cyano, halogen, hydroxyl and -C(O)H.

[0070] Example 34. The method as described in any one of Examples 23-33, wherein the AhR antagonist is any one of those listed in Table 2 or a pharmaceutically acceptable salt thereof.

[0071] Example 35. An in vitro method for predicting the responsiveness of a patient’s cancer to immune checkpoint inhibitor (ICI) therapy, comprising: (a) measuring or having measured the amount of at least one aryl hydrocarbon receptor (AhR) ligand from a serum, plasma, or fecal sample obtained from the patient; (b) comparing or having compared the amount of the at least one AhR ligand to a threshold fraction; and (c) determining that the patient’s cancer may be responsive to ICI therapy if the amount of the at least one AhR ligand is equal to or less than the threshold fraction.

[0072] Example 36. The method as described in Example 35, wherein the patient has non-small cell lung cancer, small cell lung cancer, head and neck squamous cell carcinoma, renal cell carcinoma, gastric adenocarcinoma, nasopharyngeal carcinoma, urothelial carcinoma, colorectal cancer, pleural mesothelioma, triple-negative breast cancer, esophageal tumor, multiple myeloma, gastric and gastroesophageal junction cancer, melanoma, Hodgkin lymphoma, hepatocellular carcinoma, lung cancer, head and neck cancer, non-Hodgkin lymphoma, metastatic clear cell renal cell carcinoma, lung squamous cell carcinoma, mesothelioma, gastric cancer, gastroesophageal junction cancer, metastatic melanoma, metastatic non-cutaneous melanoma, Urothelial carcinoma, diffuse large B-cell lymphoma, renal cell carcinoma, ovarian cancer, fallopian tube cancer, peritoneal tumor, extensive-stage small cell lung cancer, bladder cancer, transitional cell carcinoma, prostate tumor, recurrent or metastatic PD-L1 positive or negative squamous cell carcinoma of the head and neck, recurrent squamous cell lung cancer, advanced solid malignant tumors, pharyngeal squamous cell carcinoma, laryngeal squamous cell carcinoma, unresectable or metastatic melanoma, biliary tract tumors, esophageal squamous cell carcinoma, breast cancer, pancreatic cancer, glioblastoma, metastatic cancer, prostate cancer, solid organ cancer, gastric cancer, colon cancer, or liver cancer.

[0073] Example 37. The method as described in Example 35 or Example 36, wherein the at least one AhR ligand comprises kynurenine, tryptophan, phenylpyruvic acid, indolelacic acid, or a combination thereof.

[0074] Example 38. The method as described in Example 37, wherein the at least one AhR ligand comprises kynurenine, tryptophan, phenylpyruvic acid, and indolelacic acid.

[0075] Example 39. The method as described in any one of Examples 35-38, wherein the amount of the at least one AhR ligand in the sample is measured by liquid chromatography-mass spectrometry or gas chromatography-mass spectrometry.

[0076] Example 40. The method as described in any one of Examples 35-39, wherein the ICI therapy comprises an anti-PD-1 antibody, an anti-PD-L1 antibody, an anti-CTLA-4 antibody, an anti-LAG-3 antibody, an anti-TIM-3 antibody, an anti-TIGIT antibody, an anti-VISTA antibody, or a combination thereof.

[0077] Example 41. The method as described in any one of Examples 35-40, wherein the threshold fraction is determined by measuring the amount of at least one corresponding AhR ligand in a control serum, plasma, or fecal sample of a patient who has responded to ICI therapy.

[0078] Example 42. The method as described in any one of Examples 35-41, further comprising determining that the patient’s cancer may be responsive to ICI therapy in combination with an AhR antagonist if the amount of the at least one AhR ligand is above a threshold fraction.

[0079] Example 43. The method as described in Example 42, wherein the AhR antagonist is a compound of formula I:

[0080] Or its pharmaceutically acceptable salt.

[0081] And among them:

[0082] R 1 and R 2 Each of these is independently selected from optionally substituted alkyl groups, optionally substituted ester groups, optionally substituted heteroalkyl groups, optionally substituted acyl groups, optionally substituted amide groups, optionally substituted aryl groups, optionally substituted heteroaryl groups, optionally substituted cycloalkyl groups, optionally substituted amine groups, and optionally substituted heterocycloalkyl groups.

[0083] R 3 The group is selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, optionally substituted amide, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted ester, optionally substituted heteroalkyl, optionally substituted heteroaryl, optionally substituted heterocycloalkyl, optionally substituted amine, cyano, halogen, hydroxyl and -C(O)H.

[0084] Example 44. The method as described in Example 42 or Example 43, wherein the AhR antagonist is a compound of formula Ia:

[0085] Or its pharmaceutically acceptable salt.

[0086] And among them:

[0087] Ring A is selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, and optionally substituted heterocyclic alkyl.

[0088] Ring B is selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, and optionally substituted heterocycloalkyl.

[0089] R is selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, optionally substituted amide, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted ester, optionally substituted heteroalkyl, optionally substituted heteroaryl, optionally substituted heterocycloalkyl, amino, cyano, halogen, hydroxyl and -C(O)H.

[0090] Example 45. The method as described in any one of Examples 42-44, wherein the AhR antagonist is any one of those listed in Table 2 or a pharmaceutically acceptable salt thereof.

[0091] Example 46. A method of treating cancer in a patient in need, comprising treating the patient with an effective amount of an aryl hydrocarbon receptor (AhR) antagonist, wherein at least one AhR ligand is obtained from a serum, plasma, or fecal sample of the patient in an amount exceeding a threshold fraction.

[0092] Example 47. The method as described in Example 46, wherein the patient has non-small cell lung cancer, small cell lung cancer, head and neck squamous cell carcinoma, renal cell carcinoma, gastric adenocarcinoma, nasopharyngeal carcinoma, urothelial carcinoma, colorectal cancer, pleural mesothelioma, triple-negative breast cancer, esophageal tumor, multiple myeloma, gastric and gastroesophageal junction cancer, melanoma, Hodgkin lymphoma, hepatocellular carcinoma, lung cancer, head and neck cancer, non-Hodgkin lymphoma, metastatic clear cell renal cell carcinoma, lung squamous cell carcinoma, mesothelioma, gastric cancer, gastroesophageal junction cancer, metastatic melanoma, metastatic non-cutaneous melanoma, Urothelial carcinoma, diffuse large B-cell lymphoma, renal cell carcinoma, ovarian cancer, fallopian tube cancer, peritoneal tumor, extensive-stage small cell lung cancer, bladder cancer, transitional cell carcinoma, prostate tumor, recurrent or metastatic PD-L1 positive or negative squamous cell carcinoma of the head and neck, recurrent squamous cell lung cancer, advanced solid malignant tumors, pharyngeal squamous cell carcinoma, laryngeal squamous cell carcinoma, unresectable or metastatic melanoma, biliary tract tumors, esophageal squamous cell carcinoma, breast cancer, pancreatic cancer, glioblastoma, metastatic cancer, prostate cancer, solid organ cancer, gastric cancer, colon cancer, or liver cancer.

[0093] Example 48. The method as described in Example 46 or Example 47, wherein the patient has been treated with immune checkpoint inhibitor (ICI) therapy but is unresponsive to ICI therapy.

[0094] Example 49. The method as described in Example 46 or Example 47, wherein the patient has been treated with ICI therapy and has developed resistance to ICI therapy.

[0095] Example 50. The method as described in any one of Examples 46-49, wherein the at least one AhR ligand comprises kynurenine, tryptophan, phenylpyruvic acid, indolelacic acid, or a combination thereof.

[0096] Example 51. The method as described in Example 50, wherein the at least one AhR ligand comprises kynurenine, tryptophan, phenylpyruvic acid, and indolelacic acid.

[0097] Example 52. The method as described in any one of Examples 46-51, wherein the amount of the at least one AhR ligand in the sample is measured by liquid chromatography-mass spectrometry or gas chromatography-mass spectrometry.

[0098] Example 53. The method as described in any one of Examples 46-52, wherein the threshold fraction is determined by measuring the amount of at least one corresponding AhR ligand in a control serum, plasma, or fecal sample of a patient who has responded to ICI therapy.

[0099] Example 54. The method as described in any one of Examples 46-53, wherein the AhR antagonist is a compound of Formula I:

[0100] Or its pharmaceutically acceptable salt.

[0101] And among them:

[0102] R 1 and R 2 Each of these is independently selected from optionally substituted alkyl groups, optionally substituted ester groups, optionally substituted heteroalkyl groups, optionally substituted acyl groups, optionally substituted amide groups, optionally substituted aryl groups, optionally substituted heteroaryl groups, optionally substituted cycloalkyl groups, optionally substituted amine groups, and optionally substituted heterocycloalkyl groups.

[0103] R 3 The group is selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, optionally substituted amide, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted ester, optionally substituted heteroalkyl, optionally substituted heteroaryl, optionally substituted heterocycloalkyl, optionally substituted amine, cyano, halogen, hydroxyl and -C(O)H.

[0104] Example 55. The method as described in any one of Examples 46-53, wherein the AhR antagonist is a compound of formula Ia:

[0105] Or its pharmaceutically acceptable salt.

[0106] And among them:

[0107] Ring A is selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, and optionally substituted heterocyclic alkyl.

[0108] Ring B is selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, and optionally substituted heterocycloalkyl.

[0109] R is selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, optionally substituted amide, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted ester, optionally substituted heteroalkyl, optionally substituted heteroaryl, optionally substituted heterocycloalkyl, amino, cyano, halogen, hydroxyl and -C(O)H.

[0110] Example 56. The method as described in any one of Examples 46-55, wherein the AhR antagonist is any one of those listed in Table 2 or a pharmaceutically acceptable salt thereof.

[0111] Example 57. A method of treating cancer in a patient in need, comprising treating the patient with a combination therapy comprising an immune checkpoint inhibitor (ICI) and an aryl hydrocarbon receptor (AhR) antagonist, wherein at least one AhR ligand is obtained from a serum, plasma, or fecal sample of the patient in an amount exceeding a threshold fraction.

[0112] Example 58. The method as described in Example 57, wherein the patient has non-small cell lung cancer, small cell lung cancer, head and neck squamous cell carcinoma, renal cell carcinoma, gastric adenocarcinoma, nasopharyngeal carcinoma, urothelial carcinoma, colorectal cancer, pleural mesothelioma, triple-negative breast cancer, esophageal tumor, multiple myeloma, gastric and gastroesophageal junction cancer, melanoma, Hodgkin lymphoma, hepatocellular carcinoma, lung cancer, head and neck cancer, non-Hodgkin lymphoma, metastatic clear cell renal cell carcinoma, lung squamous cell carcinoma, mesothelioma, gastric cancer, gastroesophageal junction cancer, metastatic melanoma, metastatic non-cutaneous melanoma, Urothelial carcinoma, diffuse large B-cell lymphoma, renal cell carcinoma, ovarian cancer, fallopian tube cancer, peritoneal tumor, extensive-stage small cell lung cancer, bladder cancer, transitional cell carcinoma, prostate tumor, recurrent or metastatic PD-L1 positive or negative squamous cell carcinoma of the head and neck, recurrent squamous cell lung cancer, advanced solid malignant tumors, pharyngeal squamous cell carcinoma, laryngeal squamous cell carcinoma, unresectable or metastatic melanoma, biliary tract tumors, esophageal squamous cell carcinoma, breast cancer, pancreatic cancer, glioblastoma, metastatic cancer, prostate cancer, solid organ cancer, gastric cancer, colon cancer, or liver cancer.

[0113] Example 59. The method as described in Example 57 or Example 58, wherein the patient has been treated with immune checkpoint inhibitor (ICI) therapy but is unresponsive to ICI therapy.

[0114] Example 60. The method as described in Example 57 or Example 58, wherein the patient has been treated with ICI therapy and has developed resistance to ICI therapy.

[0115] Example 61. The method as described in any one of Examples 57-60, wherein the at least one AhR ligand comprises kynurenine, tryptophan, phenylpyruvic acid, indolelacic acid, or a combination thereof.

[0116] Example 62. The method as described in Example 61, wherein the at least one AhR ligand comprises kynurenine, tryptophan, phenylpyruvic acid, and indolelacic acid.

[0117] Example 63. The method as described in any one of Examples 57-62, wherein the amount of the at least one AhR ligand in the sample is measured by liquid chromatography-mass spectrometry or gas chromatography-mass spectrometry.

[0118] Example 64. The method as described in any one of Examples 57-63, wherein the ICI therapy comprises an anti-PD-1 antibody, an anti-PD-L1 antibody, an anti-CTLA-4 antibody, an anti-LAG-3 antibody, an anti-TIM-3 antibody, an anti-TIGIT antibody, an anti-VISTA antibody, or a combination thereof.

[0119] Example 65. The method as described in any one of Examples 57-64, wherein the threshold fraction is determined by measuring the amount of at least one corresponding AhR ligand in a control serum, plasma, or fecal sample of a patient who has responded to ICI therapy.

[0120] Example 66. The method as described in any one of Examples 57-65, wherein the AhR antagonist is a compound of Formula I:

[0121] Or its pharmaceutically acceptable salt.

[0122] And among them:

[0123] R 1 and R 2 Each of these is independently selected from optionally substituted alkyl groups, optionally substituted ester groups, optionally substituted heteroalkyl groups, optionally substituted acyl groups, optionally substituted amide groups, optionally substituted aryl groups, optionally substituted heteroaryl groups, optionally substituted cycloalkyl groups, optionally substituted amine groups, and optionally substituted heterocycloalkyl groups.

[0124] R 3The group is selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, optionally substituted amide, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted ester, optionally substituted heteroalkyl, optionally substituted heteroaryl, optionally substituted heterocycloalkyl, optionally substituted amine, cyano, halogen, hydroxyl and -C(O)H.

[0125] Example 67. The method as described in any one of Examples 57-66, wherein the AhR antagonist is a compound of formula Ia:

[0126] Or its pharmaceutically acceptable salt.

[0127] And among them:

[0128] Ring A is selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, and optionally substituted heterocyclic alkyl.

[0129] Ring B is selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, and optionally substituted heterocycloalkyl.

[0130] R is selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, optionally substituted amide, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted ester, optionally substituted heteroalkyl, optionally substituted heteroaryl, optionally substituted heterocycloalkyl, amino, cyano, halogen, hydroxyl and -C(O)H.

[0131] Example 68. The method as described in any one of Examples 57-66, wherein the AhR antagonist is any one of those listed in Table 2 or a pharmaceutically acceptable salt thereof.

[0132] Example 69. An aryl hydrocarbon receptor (AhR) antagonist for use in a method of treating cancer in patients in need, wherein at least one AhR ligand is obtained from a patient’s serum, plasma, or fecal sample in an amount exceeding a threshold fraction.

[0133] Example 70. An AhR antagonist for use as described in Example 69, wherein the patient has non-small cell lung cancer, small cell lung cancer, head and neck squamous cell carcinoma, renal cell carcinoma, gastric adenocarcinoma, nasopharyngeal carcinoma, urothelial carcinoma, colorectal cancer, pleural mesothelioma, triple-negative breast cancer, esophageal tumor, multiple myeloma, gastric and gastroesophageal junction cancer, melanoma, Hodgkin lymphoma, hepatocellular carcinoma, lung cancer, head and neck cancer, non-Hodgkin lymphoma, metastatic clear cell renal cell carcinoma, lung squamous cell carcinoma, mesothelioma, gastric cancer, gastroesophageal junction cancer, metastatic melanoma, metastatic non-scutellariae carcinoma. Skin melanoma, urothelial carcinoma, diffuse large B-cell lymphoma, renal cell carcinoma, ovarian cancer, fallopian tube cancer, peritoneal tumor, extensive-stage small cell lung cancer, bladder cancer, transitional cell carcinoma, prostate tumor, recurrent or metastatic PD-L1 positive or negative squamous cell carcinoma of the head and neck, recurrent squamous cell lung cancer, advanced solid malignant tumors, pharyngeal squamous cell carcinoma, laryngeal squamous cell carcinoma, unresectable or metastatic melanoma, biliary tract tumors, esophageal squamous cell carcinoma, breast cancer, pancreatic cancer, glioblastoma, metastatic cancer, prostate cancer, solid organ cancer, gastric cancer, colon cancer, or liver cancer.

[0134] Example 71. An AhR antagonist for use as described in Example 69 or Example 70, wherein the patient has been treated with immune checkpoint inhibitor (ICI) therapy but has not responded to ICI therapy.

[0135] Example 72. An AhR antagonist for use as described in Example 69 or Example 70, wherein the patient has been treated with ICI therapy and has developed resistance to ICI therapy.

[0136] Example 73. An AhR antagonist for use as described in any one of Examples 69-72, wherein the at least one AhR ligand comprises kynurenine, tryptophan, phenylpyruvic acid, indolelacic acid, or a combination thereof.

[0137] Example 74. An AhR antagonist for use as described in Example 73, wherein the at least one AhR ligand comprises kynurenine, tryptophan, phenylpyruvic acid, and indolelacic acid.

[0138] Example 75. An AhR antagonist for use as described in any one of Examples 69-74, wherein the amount of the at least one AhR ligand in the sample is measured by liquid chromatography-mass spectrometry or gas chromatography-mass spectrometry.

[0139] Example 76. An AhR antagonist for use as described in any one of Examples 69-75, wherein the threshold fraction is determined by measuring the amount of at least one corresponding AhR ligand in a control serum, plasma, or fecal sample obtained from a patient responding to ICI therapy.

[0140] Example 77. An AhR antagonist for use as described in any one of Examples 69-76, wherein the AhR antagonist is a compound of Formula I:

[0141] Or its pharmaceutically acceptable salt.

[0142] And among them:

[0143] R 1 and R 2 Each of these is independently selected from optionally substituted alkyl groups, optionally substituted ester groups, optionally substituted heteroalkyl groups, optionally substituted acyl groups, optionally substituted amide groups, optionally substituted aryl groups, optionally substituted heteroaryl groups, optionally substituted cycloalkyl groups, optionally substituted amine groups, and optionally substituted heterocycloalkyl groups.

[0144] R 3 The group is selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, optionally substituted amide, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted ester, optionally substituted heteroalkyl, optionally substituted heteroaryl, optionally substituted heterocycloalkyl, optionally substituted amine, cyano, halogen, hydroxyl and -C(O)H.

[0145] Example 78. An AhR antagonist for use as described in any one of Examples 69-77, wherein the AhR antagonist is a compound of formula Ia:

[0146] Or its pharmaceutically acceptable salt.

[0147] And among them:

[0148] Ring A is selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, and optionally substituted heterocyclic alkyl.

[0149] Ring B is selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, and optionally substituted heterocycloalkyl.

[0150] R is selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, optionally substituted amide, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted ester, optionally substituted heteroalkyl, optionally substituted heteroaryl, optionally substituted heterocycloalkyl, amino, cyano, halogen, hydroxyl and -C(O)H.

[0151] Example 79. An AhR antagonist for use as described in any one of Examples 69-77, wherein the AhR antagonist is any one of those listed in Table 2 or a pharmaceutically acceptable salt thereof.

[0152] Example 80. A combination therapy for use in a method of treating cancer in a patient in need, wherein the combination therapy comprises an immune checkpoint inhibitor (ICI) and an aryl hydrocarbon receptor (AhR) antagonist, and further wherein at least one AhR ligand is obtained from a serum, plasma, or fecal sample of the patient in an amount exceeding a threshold fraction.

[0153] Example 81. A combination therapy for use as described in Example 80, wherein the patient has non-small cell lung cancer, small cell lung cancer, head and neck squamous cell carcinoma, renal cell carcinoma, gastric adenocarcinoma, nasopharyngeal carcinoma, urothelial carcinoma, colorectal cancer, pleural mesothelioma, triple-negative breast cancer, esophageal tumor, multiple myeloma, gastric and gastroesophageal junction cancer, melanoma, Hodgkin lymphoma, hepatocellular carcinoma, lung cancer, head and neck cancer, non-Hodgkin lymphoma, metastatic clear cell renal cell carcinoma, lung squamous cell carcinoma, mesothelioma, gastric cancer, gastroesophageal junction cancer, metastatic melanoma, metastatic non-cutaneous carcinoma. Melanoma, urothelial carcinoma, diffuse large B-cell lymphoma, renal cell carcinoma, ovarian cancer, fallopian tube cancer, peritoneal tumor, extensive-stage small cell lung cancer, bladder cancer, transitional cell carcinoma, prostate cancer, recurrent or metastatic PD-L1 positive or negative squamous cell carcinoma of the head and neck, recurrent squamous cell lung cancer, advanced solid malignant tumors, pharyngeal squamous cell carcinoma, laryngeal squamous cell carcinoma, unresectable or metastatic melanoma, biliary tract tumors, esophageal squamous cell carcinoma, breast cancer, pancreatic cancer, glioblastoma, metastatic cancer, prostate cancer, solid organ cancer, gastric cancer, colon cancer, or liver cancer.

[0154] Example 82. A combination therapy for use as described in Example 80 or Example 81, wherein the patient has been treated with immune checkpoint inhibitor (ICI) therapy but has not responded to ICI therapy.

[0155] Example 83. A combination therapy for use as described in Example 80 or Example 81, wherein the patient has been treated with ICI therapy and has developed resistance to ICI therapy.

[0156] Example 84. A combination therapy for use as described in any one of Examples 80-83, wherein the at least one AhR ligand comprises kynurenine, tryptophan, phenylpyruvic acid, indolelacic acid, or a combination thereof.

[0157] Example 85. A combination therapy for use as described in Example 84, wherein the at least one AhR ligand comprises kynurenine, tryptophan, phenylpyruvic acid, and indolelactate.

[0158] Example 86. A combination therapy for use as described in any one of Examples 80-85, wherein the amount of the at least one AhR ligand in the sample is measured by liquid chromatography-mass spectrometry or gas chromatography-mass spectrometry.

[0159] Example 87. A combination therapy for use as described in any one of Examples 80-86, wherein the ICI therapy comprises an anti-PD-1 antibody, an anti-PD-L1 antibody, an anti-CTLA-4 antibody, an anti-LAG-3 antibody, an anti-TIM-3 antibody, an anti-TIGIT antibody, an anti-VISTA antibody, or a combination thereof.

[0160] Example 88. A combination therapy for use as described in any one of Examples 80-87, wherein the threshold fraction is determined by measuring the amount of at least one corresponding AhR ligand in a control serum, plasma, or fecal sample of a patient who has responded to ICI therapy.

[0161] Example 89. A combination therapy for use as described in any one of Examples 80-88, wherein the AhR antagonist is a compound of Formula I:

[0162] Or its pharmaceutically acceptable salt.

[0163] And among them:

[0164] R 1 and R 2 Each of these is independently selected from optionally substituted alkyl groups, optionally substituted ester groups, optionally substituted heteroalkyl groups, optionally substituted acyl groups, optionally substituted amide groups, optionally substituted aryl groups, optionally substituted heteroaryl groups, optionally substituted cycloalkyl groups, optionally substituted amine groups, and optionally substituted heterocycloalkyl groups.

[0165] R 3 The group is selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, optionally substituted amide, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted ester, optionally substituted heteroalkyl, optionally substituted heteroaryl, optionally substituted heterocycloalkyl, optionally substituted amine, cyano, halogen, hydroxyl and -C(O)H.

[0166] Example 90. A combination therapy for use as described in any one of Examples 80-89, wherein the AhR antagonist is a compound of formula Ia:

[0167] Or its pharmaceutically acceptable salt.

[0168] And among them:

[0169] Ring A is selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, and optionally substituted heterocyclic alkyl.

[0170] Ring B is selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, and optionally substituted heterocycloalkyl.

[0171] R is selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, optionally substituted amide, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted ester, optionally substituted heteroalkyl, optionally substituted heteroaryl, optionally substituted heterocycloalkyl, amino, cyano, halogen, hydroxyl and -C(O)H.

[0172] Example 91. A combination therapy for use as described in any one of Examples 80-89, wherein the AhR antagonist is any one listed in Table 2 or a pharmaceutically acceptable salt thereof.

[0173] Example 92. Use of an aryl hydrocarbon receptor (AhR) antagonist in the manufacture of a medicament for treating cancer in patients in need, wherein at least one AhR ligand is obtained from a patient’s serum, plasma, or fecal sample in an amount exceeding a threshold fraction.

[0174] Example 93. Use as described in Example 92, wherein the patient has non-small cell lung cancer, small cell lung cancer, head and neck squamous cell carcinoma, renal cell carcinoma, gastric adenocarcinoma, nasopharyngeal carcinoma, urothelial carcinoma, colorectal cancer, pleural mesothelioma, triple-negative breast cancer, esophageal tumor, multiple myeloma, gastric and gastroesophageal junction cancer, melanoma, Hodgkin lymphoma, hepatocellular carcinoma, lung cancer, head and neck cancer, non-Hodgkin lymphoma, metastatic clear cell renal cell carcinoma, lung squamous cell carcinoma, mesothelioma, gastric cancer, gastroesophageal junction cancer, metastatic melanoma, metastatic non-cutaneous melanoma. Urothelial carcinoma, diffuse large B-cell lymphoma, renal cell carcinoma, ovarian cancer, fallopian tube cancer, peritoneal tumor, extensive-stage small cell lung cancer, bladder cancer, transitional cell carcinoma, prostate tumor, recurrent or metastatic PD-L1 positive or negative squamous cell carcinoma of the head and neck, recurrent squamous cell lung cancer, advanced solid malignant tumors, pharyngeal squamous cell carcinoma, laryngeal squamous cell carcinoma, unresectable or metastatic melanoma, biliary tract tumors, esophageal squamous cell carcinoma, breast cancer, pancreatic cancer, glioblastoma, metastatic cancer, prostate cancer, solid organ cancer, gastric cancer, colon cancer, or liver cancer.

[0175] Example 94. Use as described in Example 92 or Example 93, wherein the patient has been treated with immune checkpoint inhibitor (ICI) therapy but has not responded to ICI therapy.

[0176] Example 95. Use as described in Example 92 or Example 93, wherein the patient has been treated with ICI therapy and has developed resistance to ICI therapy.

[0177] Example 96. Use as described in any one of Examples 92-95, wherein the at least one AhR ligand comprises kynurenine, tryptophan, phenylpyruvic acid, indolelacic acid, or a combination thereof.

[0178] Example 97. Use as described in Example 96, wherein the at least one AhR ligand comprises kynurenine, tryptophan, phenylpyruvic acid, and indolelacic acid.

[0179] Example 98. Use as described in any one of Examples 92-97, wherein the amount of the at least one AhR ligand in the sample is measured by liquid chromatography-mass spectrometry or gas chromatography-mass spectrometry.

[0180] Example 99. Use as described in any one of Examples 92-98, wherein the threshold fraction is determined by measuring the amount of at least one corresponding AhR ligand in a control serum, plasma, or fecal sample of a patient who has responded to ICI therapy.

[0181] Example 100. Use as described in any one of Examples 92-99, wherein the AhR antagonist is a compound of Formula I:

[0182] Or its pharmaceutically acceptable salt.

[0183] And among them:

[0184] R 1 and R 2 Each of these is independently selected from optionally substituted alkyl groups, optionally substituted ester groups, optionally substituted heteroalkyl groups, optionally substituted acyl groups, optionally substituted amide groups, optionally substituted aryl groups, optionally substituted heteroaryl groups, optionally substituted cycloalkyl groups, optionally substituted amine groups, and optionally substituted heterocycloalkyl groups.

[0185] R 3 The group is selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, optionally substituted amide, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted ester, optionally substituted heteroalkyl, optionally substituted heteroaryl, optionally substituted heterocycloalkyl, optionally substituted amine, cyano, halogen, hydroxyl and -C(O)H.

[0186] Example 101. Use as described in any one of Examples 92-100, wherein the AhR antagonist is a compound of formula Ia:

[0187] Or its pharmaceutically acceptable salt.

[0188] And among them:

[0189] Ring A is selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, and optionally substituted heterocyclic alkyl.

[0190] Ring B is selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, and optionally substituted heterocycloalkyl.

[0191] R is selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, optionally substituted amide, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted ester, optionally substituted heteroalkyl, optionally substituted heteroaryl, optionally substituted heterocycloalkyl, amino, cyano, halogen, hydroxyl and -C(O)H.

[0192] Example 102. Use as described in any one of Examples 92-100, wherein the AhR antagonist is any one of those listed in Table 2 or a pharmaceutically acceptable salt thereof.

[0193] Example 103. Use of immune checkpoint inhibitors (ICIs) and aryl hydrocarbon receptor (AhR) antagonists in the manufacture of medicaments for the treatment of cancer in patients in need, wherein at least one AhR ligand is obtained from a patient’s serum, plasma, or fecal sample in an amount exceeding a threshold fraction.

[0194] Example 104. Use as described in Example 103, wherein the patient has non-small cell lung cancer, small cell lung cancer, head and neck squamous cell carcinoma, renal cell carcinoma, gastric adenocarcinoma, nasopharyngeal carcinoma, urothelial carcinoma, colorectal cancer, pleural mesothelioma, triple-negative breast cancer, esophageal tumor, multiple myeloma, gastric and gastroesophageal junction cancer, melanoma, Hodgkin lymphoma, hepatocellular carcinoma, lung cancer, head and neck cancer, non-Hodgkin lymphoma, metastatic clear cell renal cell carcinoma, lung squamous cell carcinoma, mesothelioma, gastric cancer, gastroesophageal junction cancer, metastatic melanoma, or metastatic non-cutaneous melanoma. Urothelial carcinoma, diffuse large B-cell lymphoma, renal cell carcinoma, ovarian cancer, fallopian tube cancer, peritoneal tumors, extensive-stage small cell lung cancer, bladder cancer, transitional cell carcinoma, prostate cancer, recurrent or metastatic PD-L1 positive or negative squamous cell carcinoma of the head and neck, recurrent squamous cell lung cancer, advanced solid malignant tumors, pharyngeal squamous cell carcinoma, laryngeal squamous cell carcinoma, unresectable or metastatic melanoma, biliary tract tumors, esophageal squamous cell carcinoma, breast cancer, pancreatic cancer, glioblastoma, metastatic cancer, prostate cancer, solid organ cancer, gastric cancer, colon cancer, or liver cancer.

[0195] Example 105. Use as described in Example 103 or Example 104, wherein the patient has been treated with immune checkpoint inhibitor (ICI) therapy but has not responded to ICI therapy.

[0196] Example 106. Use as described in Example 103 or Example 104, wherein the patient has been treated with ICI therapy and has developed resistance to ICI therapy.

[0197] Example 107. Use as described in any one of Examples 103-106, wherein the at least one AhR ligand comprises kynurenine, tryptophan, phenylpyruvic acid, indolelacic acid, or a combination thereof.

[0198] Example 108. Use as described in Example 107, wherein the at least one AhR ligand comprises kynurenine, tryptophan, phenylpyruvic acid and indolelacic acid.

[0199] Example 109. Use as described in any one of Examples 103-108, wherein the amount of the at least one AhR ligand in the sample is measured by liquid chromatography-mass spectrometry or gas chromatography-mass spectrometry.

[0200] Example 110. Use as described in any one of Examples 103-109, wherein the ICI therapy comprises an anti-PD-1 antibody, an anti-PD-L1 antibody, an anti-CTLA-4 antibody, an anti-LAG-3 antibody, an anti-TIM-3 antibody, an anti-TIGIT antibody, an anti-VISTA antibody, or a combination thereof.

[0201] Example 111. Use as described in any one of Examples 103-110, wherein the threshold fraction is determined by measuring the amount of at least one corresponding AhR ligand in a control serum, plasma, or fecal sample of a patient who has responded to ICI therapy.

[0202] Example 112. Use as described in any one of Examples 103-111, wherein the AhR antagonist is a compound of Formula I:

[0203] Or its pharmaceutically acceptable salt.

[0204] And among them:

[0205] R 1 and R 2 Each of these is independently selected from optionally substituted alkyl groups, optionally substituted ester groups, optionally substituted heteroalkyl groups, optionally substituted acyl groups, optionally substituted amide groups, optionally substituted aryl groups, optionally substituted heteroaryl groups, optionally substituted cycloalkyl groups, optionally substituted amine groups, and optionally substituted heterocycloalkyl groups.

[0206] R 3 The group is selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, optionally substituted amide, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted ester, optionally substituted heteroalkyl, optionally substituted heteroaryl, optionally substituted heterocycloalkyl, optionally substituted amine, cyano, halogen, hydroxyl and -C(O)H.

[0207] Example 113. Use as described in any one of Examples 103-112, wherein the AhR antagonist is a compound of formula Ia:

[0208] Or its pharmaceutically acceptable salt.

[0209] And among them:

[0210] Ring A is selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, and optionally substituted heterocyclic alkyl.

[0211] Ring B is selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, and optionally substituted heterocycloalkyl.

[0212] R is selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, optionally substituted amide, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted ester, optionally substituted heteroalkyl, optionally substituted heteroaryl, optionally substituted heterocycloalkyl, amino, cyano, halogen, hydroxyl and -C(O)H.

[0213] Example 114. Use as described in any one of Examples 103-112, wherein the AhR antagonist is any one of those listed in Table 2 or a pharmaceutically acceptable salt thereof. Attached Figure Description

[0214] Figure 1A-1B The following are fecal samples obtained from 73 patients in Example 1. Figure 1A ) and plasma samples ( Figure 1B The levels of kynurenine, tryptophan, phenylpyruvic acid, and indolelacic acid (collectively referred to as relevant ligands) in these patients were eventually characterized as either responders or non-responders.

[0215] Figure 2 Elevated levels of relevant ligands were shown from stool samples obtained from 19 patients who received ICI therapy alone (without concomitant therapy) and who were eventually characterized as either responders or non-responders.

[0216] Figure 3 Principal component analysis shows the reduced signal dimensionality from all 29 AhR ligands compared to their associated ligands. Left panel: 73 patients - 29 AhR ligands; Middle panel: 73 patients - associated ligands; Right panel: 19 patients who received ICI therapy only - associated ligands.

[0217] Figures 4A-4B The following are tumor growth curves measured in a mouse model of colon cancer resistant to anti-PD-L1 therapy after administration of an isotope control antibody, an anti-PD-L1 antibody, and compound number 30 from Table 2 combined with an anti-PD-L1 antibody (BioXcell anti-mouse PD-L1, B7-H1 clone 10F.9G2). Figure 4A ) and tumor weight ( Figure 4B ).

[0218] Figure 5The levels of AhR ligands described in Table 1 are shown in patients who received ICI therapy alone (without concomitant therapy) and who were eventually characterized as responders or non-responders.

[0219] This disclosure is based in part on the finding that certain AhR ligands, such as kynurenine, tryptophan, phenylpyruvate, and indolelactone, are available biomarkers for predicting a patient’s cancer response to ICI therapy. In some embodiments, this disclosure provides a method for predicting a patient’s cancer response to ICI therapy based on determining the amount of at least one biomarker relative to a threshold fraction. In some embodiments, the biomarkers described herein can enable the identification and / or stratification of cancer patients who will benefit from treatment with an AhR antagonist. In some embodiments, the biomarkers described herein can be used to inform and provide effective treatment options to cancer patients undergoing one or more ICI therapies.

[0220] It should be understood that treatments using aryl hydrocarbon receptor (AhR) antagonists or combination therapies (e.g., cancer treatments) mentioned in this article should also be interpreted as mentions:

[0221] -AhR antagonists or combination therapies for use in treatments such as cancer; and / or

[0222] -AhR antagonists or combination therapies are used in the manufacture of drugs for treating, for example, cancer.

[0223] The following detailed description and examples illustrate certain embodiments of this disclosure. Those skilled in the art will recognize that the scope of this disclosure includes many variations and modifications. Therefore, the description of certain embodiments should not be construed as limiting the scope of this disclosure.

[0224] definition

[0225] To facilitate understanding of this disclosure, certain terms are defined throughout the detailed description. Unless otherwise defined herein, all scientific and technical terms used in conjunction with this disclosure have the same meaning as commonly understood by one of ordinary skill in the art.

[0226] As used herein, unless the context clearly indicates otherwise, the singular form of a word includes the plural form; for example, the terms “a / an” and “the” should be understood as either singular or plural. Unless the specific context clearly indicates otherwise, the term “or” should mean “and / or”.

[0227] As used herein, the term "biomarker" refers to a biological compound that is present in a biological sample and can be isolated from or measured in a biological sample. Such biomarkers include, but are not limited to, nucleic acids, proteins, carbohydrates, lipids, organic or inorganic chemicals, natural polymers, and metabolites. In some embodiments, a biomarker includes one or more AhR ligands, such as kynurenine, tryptophan, phenylpyruvic acid, and indolelactate, as shown in Example 1. In some embodiments, biomarkers according to this disclosure can predict the response of cancer patients to ICI therapy.

[0228] As used herein, the terms “measurement” or “detection” refer to assessing the presence, absence, or amount of a biomarker disclosed herein in a biological sample, including the derivation of a quantitative level of such biomarker. In some embodiments, the amount of at least one biomarker in a biological sample is measured or detected using methods known in the art, such as liquid chromatography-mass spectrometry and gas chromatography-mass spectrometry.

[0229] As used herein, the term "threshold score" refers to a reference biomarker amount suitable for providing a comparison with the amount of the biomarker in a test sample. In some embodiments, the threshold score is the amount of a biomarker from a serum, plasma, or stool sample of a cancer patient responding to ICI therapy. In some embodiments, the threshold score may be generated from a population or cohort of two or more cancer patients responding to ICI therapy. A population or cohort may, for example, include at least 2, 3, 4, 5, 10, 15, 18, 20, 30, 40, 50, 75, 100, or more cancer patients. In such embodiments, the threshold score for each biomarker may be the average amount of the biomarker from serum, plasma, or stool samples of a population or cohort of cancer patients responding to ICI therapy.

[0230] The terms "biological sample" and "sample" are used interchangeably and refer to a sample obtained from a patient, including bodily fluids, body tissues (e.g., tumor tissue), cells, or other sources. In some embodiments, a biological sample includes stool obtained from a cancer patient who has undergone ICI therapy. In some embodiments, a biological sample includes plasma obtained from a cancer patient who has undergone ICI therapy. In some embodiments, a biological sample includes stool obtained from a cancer patient who has not yet received ICI therapy. In some embodiments, a biological sample includes plasma obtained from a cancer patient who has not yet received ICI therapy. Methods for obtaining such biological samples from cancer patients are known in the art.

[0231] As used herein, the term "AhR ligand" refers to a compound that binds to and activates aryl hydrocarbon receptors (i.e., AhR) expressed in many cells of the immune system, and upon binding, causes AhR to translocate to the nucleus, where it interacts with dioxin-response elements (DREs) of AhR-responsive genes to regulate their transcription. AhR ligands are known in the art and include typical xenobiotics such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and 3-methylcholanthrene. Other examples of AhR ligands include kynurenine, tryptophan, phenylpyruvic acid, indolelacic acid, β-naphthylflavonoid (BNF), 6-formylindolano(3,2-b)carbazole (FICZ), and methyl 2-(1H-indol-3-ylcarbonyl)-4-thiazolylcarboxylate (ITE). In addition, Table 1 lists the 29 AhR ligands studied in Example 1, including kynurenine, tryptophan, phenylpyruvic acid, and indolelacic acid.

[0232] As used herein, the term "AhR antagonist" refers to any agent capable of inhibiting or reducing the biological activity of the AhR it binds to. In some embodiments, the AhR antagonist reduces the activity of AhR by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 99% relative to the activity of AhR in the absence of the AhR antagonist. Certain AhR antagonists, such as α-naphthylflavonoids and resveratrol, are known in the art. Other AhR antagonists are described in this disclosure. See, for example, Table 2.

[0233] The terms “comprise,” “contain,” “have,” and “include” are open-ended connecting verbs. Any form or tense of one or more of these verbs, such as “comprise,” “comprising,” “contain,” “containing,” “has,” “having,” “include,” and “including,” are also open-ended. For example, any method that “comprises,” “contains,” “has,” or “includes” one or more steps is not limited to having only those one or more steps and may also cover other steps not listed. The use of any and all instances or exemplary language (e.g., “such as”) provided with respect to certain embodiments herein is intended only to better illustrate this disclosure and does not constitute a limitation on the scope of this disclosure as otherwise claimed.

[0234] The terms “patient” and “subject” are used interchangeably in this document and refer to a human or a non-human animal (e.g., a mammal). As used herein, the term “cancer patient” refers to a patient who has been diagnosed with cancer or a tumor by a qualified professional (e.g., a physician or nursing practitioner).

[0235] The terms “cancer” or “tumor” are used interchangeably herein and refer to cells exhibiting typical characteristics of cancerous cells, such as uncontrolled proliferation, immortality, metastatic potential, rapid growth and proliferation rates, and certain characteristic morphological features. In some embodiments, such cells exhibit these characteristics partially or completely due to the expression and activity of immune checkpoint inhibitors such as PD-1, PD-L1, and / or CTLA-4. Cancers include, but are not limited to, non-small cell lung cancer, small cell lung cancer, squamous cell carcinoma of the head and neck, renal cell carcinoma, gastric adenocarcinoma, nasopharyngeal carcinoma, urothelial carcinoma, colorectal cancer, pleural mesothelioma, triple-negative breast cancer, esophageal tumors, multiple myeloma, gastric and gastroesophageal junction cancer, melanoma, Hodgkin lymphoma, hepatocellular carcinoma, lung cancer, head and neck cancer, non-Hodgkin lymphoma, metastatic clear cell renal cell carcinoma, squamous cell carcinoma of the lung, mesothelioma, gastric cancer, gastroesophageal junction cancer, metastatic melanoma, metastatic non-cutaneous melanoma, urothelial carcinoma, diffuse Large B-cell lymphoma, renal cell carcinoma, ovarian cancer, fallopian tube cancer, peritoneal tumors, extensive-stage small cell lung cancer, bladder cancer, transitional cell carcinoma, prostate cancer, recurrent or metastatic PD-L1 positive or negative squamous cell carcinoma of the head and neck, recurrent squamous cell lung cancer, advanced solid malignancies, pharyngeal squamous cell carcinoma, laryngeal squamous cell carcinoma, unresectable or metastatic melanoma, biliary tract tumors, esophageal squamous cell carcinoma, breast cancer, pancreatic cancer, glioblastoma, metastatic cancer, prostate cancer, solid organ cancers, gastric cancer, colon cancer, and liver cancer.

[0236] As used herein, the term “immune checkpoint” refers to a group of molecules on the cell surface of CD4+ and / or CD8+ cells that downregulate or inhibit immune responses to maintain self-tolerance, prevent autoimmunity, and control the duration and extent of immune responses to minimize collateral tissue damage. Immune checkpoint proteins are known in the art and include, but are not limited to, CTLA-4, PD-1, PD-L1, VISTA, B7-H2, B7-H3, B7-H4, B7-H6, ICOS, HVEM, PD-L2, CD160, gp49B, PIR-B, KIR family receptors, TIM-1, TIM-3, TIM-4, LAG-3, GITR, 4-IBB, OX-40, BTLA, SIRP, CD47, CD48, 2B4 (CD244), B7.1, B7.2, ILT-2, ILT-4, TIGIT, HHLA2, prolactin, IDO, CD39, CD73, and A2aR.

[0237] As used herein, the term "immune checkpoint inhibitor" (ICI) refers to any therapeutic agent that inhibits one or more immune checkpoint proteins, including any small molecule chemical compound, nucleic acid molecule, or peptide (e.g., antibody), or any fragment thereof. Inhibition of one or more immune checkpoint proteins can block or otherwise neutralize inhibitory signaling, thereby upregulating the immune response for example, to treat cancer more effectively. Exemplary immune checkpoint inhibitors include antibodies against one or more immune checkpoint proteins that directly block the interaction between the protein and its natural receptor, inactive forms of one or more immune checkpoint proteins, small molecules or peptides that block the interaction between one or more immune checkpoint proteins and their natural receptors, nucleic acid molecules that block the transcription or translation of the nucleic acid of the immune checkpoint protein, etc.

[0238] As used herein, the term "treating" or "treatment" means relieving symptoms, slowing the onset or progression of symptoms, reducing the risk of disease progression, preventing or delaying the development of symptoms associated with the symptoms, reducing or terminating symptoms associated with the symptoms, producing complete or partial remission of the symptoms, curing the symptoms, or a combination thereof. In some embodiments, the condition is cancer. Regarding the treatment of cancer, the term "treating" or "treatment" may refer to providing a cancer patient with ICI therapy, optionally in combination with an AhR antagonist, resulting in improvement of at least one or more symptoms of the cancer.

[0239] As used herein, the term "effective amount" refers to an amount of a therapeutic agent or its pharmaceutically acceptable salt that is sufficient to reduce at least one or more symptoms of the conditions disclosed herein or to provide the desired effect. Regarding the treatment of cancer, "effective amount" can vary depending on factors such as the stage of cancer and / or the age, sex, and weight of the cancer patient.

[0240] As used herein, the term "administering" refers to placing a therapeutic agent (e.g., an AhR antagonist or an ICI) into a mammalian tissue or subject by means or pathways that result in the therapeutic agent being at least partially located at a desired site or tissue location. In some embodiments, the therapeutic agent may be administered to the subject using pathways known in the art, such as intramuscular injection, subcutaneous injection, and intravenous injection or infusion.

[0241] As used herein, the term "resistance" refers to acquired or natural resistance in a cancer sample or mammal to a cancer therapy (i.e., no response to the therapeutic or a reduced or limited response), such as a reduced response to the therapeutic therapy by 25% or more, for example, 30%, 40%, 50%, 60%, 70%, 80%, or more. The reduction in response can be measured by comparison with the same cancer sample or mammal before the acquisition of resistance, or by comparison with different cancer samples or mammals known to be resistant to the therapeutic therapy. The determination of resistance to a therapeutic therapy is a routine procedure in the art and is within the skill level of a general technical clinician.

[0242] The terms “respond,” “response,” or “responsiveness” refer to an anticancer response, for example, in the sense of a reduction in tumor size or inhibition of tumor growth. The term can also refer to an improved prognosis, such as reflected by an increased time to recurrence or an increased overall survival, which is the period from treatment to death from any cause. A response or having a response means achieving a beneficial endpoint upon exposure to a stimulus. Alternatively, a response or having a response means that negative or harmful symptoms are minimized, alleviated, or reduced upon exposure to a stimulus. Determining a response or responsiveness to therapeutic treatment is a routine procedure in the art and is within the skill of a general clinician. For example, whether a patient is responding to ICI therapy can be determined based on the Response Evaluation Criteria in Solid Tumors (RECIST). See Example 1.

[0243] As used herein, the term "combination therapy" refers to the delivery of two or more different treatments to a patient during a disease, disability, or condition (e.g., cancer). For example, in some embodiments, the two treatments, such as ICI therapy and an AhR antagonist, are delivered after the patient has been diagnosed with cancer and before the cancer has been cured or eliminated. In some embodiments, the delivery of one treatment still occurs when the delivery of the second treatment begins, resulting in overlap. In some embodiments, the first and second treatments begin simultaneously. In other embodiments, the first and second treatments begin at different times. In some embodiments, one treatment ends before the second treatment begins. In some embodiments, ICI therapy and an AhR antagonist are administered simultaneously. In some embodiments, ICI therapy and an AhR antagonist are administered sequentially. In some embodiments, the two treatments are administered close enough in time to provide the desired therapeutic effect.

[0244] AhR ligand as a biomarker for ICI responsiveness

[0245] According to this disclosure, certain AhR ligands, such as kynurenine, tryptophan, phenylpyruvic acid, and indolelactate, can be used as biomarkers to predict a patient’s cancer response to ICI therapy.

[0246] AhR is a ligand-activated transcription factor belonging to the basic helical-loop-helical / Per-Arnt-Sim (bHLH / PAS) protein family located in the cytosol. Upon ligand binding, AhR translocates to the nucleus, where it heterodimerizes with the AhR nuclear translocase protein (ARNT). On the ARNT, it interacts with dioxin-responsive elements (DREs) of AhR-responsive genes to regulate their transcription. Representative target genes include microsomal cytochrome P450-dependent monooxygenases, including cytochrome P450 family-1 subfamily-A polypeptide-1 (CYP1A1), cytochrome P450 family-1 subfamily-A polypeptide-2 (CYP1A2), cytochrome P450 family-1 subfamily-B polypeptide-1 (CYP1B1), and NAD(P)H-quinone oxidoreductase. AhR can also influence chromatin remodeling, act on deacetylases, and correct their gene regulatory activities to guide gene repression (Rothhammer et al., Nat RevImmunol. [Nature Immunology Reviews], 2019, 19(3):184-197). Furthermore, AhR interacts with transcription factors such as NF-κB, c-Maf, and a range of other gene regulators and influences the activities of these regulators (Hankinson et al., ArchBiochem Biophys. [Biochemistry and Biophysics Literature], 2005, 433(2):379-386). A feedback loop for NF-κB activation has also been described.

[0247] AhR is expressed in many cells of the immune system, including dendritic cells (DCs), macrophages, T cells, and NK cells, and plays a role in immune regulation (Nguyen et al., Front. Immunol., 2014, 5:551). Several reports have indicated that AhR mediates the differentiation of CD4(+) T cells, particularly Th17 and Th22, and exacerbates autoimmunity in mice (Funatake et al., J Immunol., 2005, 175(7):4184-4188; Quintana et al., Nature., 2008, 453(7191):65-71; Veldhoen et al., Nature., 2008, 453(7191):106-109; Xiong et al., Exp Cell Res., 2020, 112288). AhR also modulates the tone of adaptive immune responses and regulates the differentiation of T cells into regulatory phenotypes. In addition, Quintana et al. reported the role of AhR in regulating Th17 biology and controlling IL-22 production (Quintana et al., Nature, 2008, 453(7191):65-71; Quintana et al., Eur J Immunol, 2009, 39(3):655-657; Yeste et al., Nat Commun, 2014, 5:3753).

[0248] Under immunosuppression, AhR activation promotes regulatory T cell generation, directly and indirectly inhibits Th1 and Th17 differentiation, and reduces dendritic cell (DC) activation and maturation (Wang et al., Clin. Exp. Immunol., 2014, 177(2):521-30; Mezrich et al., J. Immunol., 2010, 185(6):3190-8; Wei et al., Lab. Invest., 2014, 94(5):528-35; Nguyen et al., PNAS, 2010, 107(46):19961-6). It is known that the classic exogenous AhR ligands TCDD and 3-methylcholanthrene induce deep immunosuppression, promote carcinogenesis, and induce tumor growth (Gramatzki et al., Oncogene, 2009, 28(28):2593-605; Bui et al., Oncogene, 2009, 28(41):3642-51; Esser et al., Trends Immunol, 2009, 30:447-454).

[0249] AhR activation also regulates the innate immune response, and constitutive AhR expression has been shown to negatively regulate the type I interferon response to viral infection (Yamada et al., Nat. Immunol. [Nature Immunology], 2016, 17(6):687-94).

[0250] Mice with constitutively active AhR spontaneously develop tumors (Andersson et al., PNAS, 2002, 99(15):9990-5). There is evidence that AhR activity is dysregulated in the tumor environment, affecting immune cells and leading to impaired immune surveillance. For example, AhR can affect tumorigenesis by directly disrupting cell proliferation, tissue invasion, angiogenesis, tumor-associated inflammation, and metastasis (Xiong et al., Experimental Cell Research, 2020, 112288; Veldhoen et al., Nature, 2008, 453(7191):106-109; Dietrich et al., Carcinogenesis, 2010, 31(8):1319-1328; Opitz et al., Nature, 2011, 478(7368):197-203).

[0251] AhR signaling can be activated by one or more AhR ligands known in the art. For example, AhR can be activated by environmental toxins such as TCDD and natural ligands provided directly by diet, or by dietary substances further converted by enzymatic activity of intestinal microsomes. These ligands, in turn, modulate the transcriptional activity of AhR in the host immune system, affecting the immune response against the microbiome. Tryptophan metabolites such as kynurenine and kynurenine have been reported to activate AhR (DiNatale et al., Toxicol. Sci., 2010, 115(1):89-97; Mezrich et al., J. Immunol., 2010, 185(6):3190-8; Opitz et al., Nature, 2011, 478(7368):197-203). Other endogenous ligands are known to bind to AHR, although their physiological effects are currently unclear (Nguyen & Bradfield, Chem. Res. Toxicol. [Chemical Research in the Field of Toxicology], 2008, 21(1):102-116).

[0252] It has been shown that the essential amino acid tryptophan is metabolized in tumor tissues by the rate-limiting enzymes indoleamine-2,3-dioxygenase 1 and 2 (IDO1 / IDO2) and tryptophan-2,3-dioxygenase 2 (TDO2) expressed in tumor cells, bone marrow, or antigen-presenting cells (Platten et al., Front Immunol. 2015, 5:673). The IDO1 / 2-mediated metabolic pathway induces an immune-allowed microenvironment in tumors and tumor-draining lymph nodes by inducing T cell unresponsiveness and apoptosis through tryptophan depletion and the accumulation of immunosuppressive tryptophan catabolites (Fallarino et al., Cell DeathDiffer., 2002, 9(10): 1069-1077; Uyttenhove et al., Nat. Med., 2003, 9(10): 1269-74; Liu et al., Blood, 2005, 115(17): 3520-30; Muller et al., Nat. Med., 11(3): 312-9; Metz, Cancer Res., 2007, 67(15): 7082-7). Competitive inhibitors of IDO are currently being tested in clinical trials in patients with solid tumors with the aim of enhancing the efficacy of conventional chemotherapy.

[0253] TDO2 is strongly expressed in cancer and can lead to the production of immunosuppressive kynurenine. In gliomas, downstream of TDO-mediated tryptophan degradation, kynurenine activation of AhR enhances tumor growth by suppressing antitumor immune responses and directly promoting tumor cell survival and motility (Opitz et al., Nature, 2011, 478(7368):197-203). Therefore, AhR ligands generated by tumor cells act on tumor cells and lymphocytes in both autocrine and paracrine modes to promote tumor growth.

[0254] Constitutive activation of the IDO1 / TDO2 / AhR pathway and nuclear AhR protein accumulation is associated with unresponsiveness to immune checkpoint inhibitors and poor overall survival (Chen et al., Medicine (Baltimore), 2020, 99(21); Ferns et al., Oncoimmunology, 2015, 4(2); Suzuki et al., Lung Cancer, 2010, 67(3):361-365). In a phase II study of melanoma, epadostat's inhibition of IDO1 improved the antitumor efficacy of the anti-PD-1 checkpoint inhibitor Keytruda, but development was terminated in phase III (ECHO-301) due to its lack of durable efficacy (Muller et al., Semin Immunopathol, 2019, 41(1):41-48). One possible explanation is that the tumor is converted to other sources of AhR ligands that are not subject to icodrostat inhibition.

[0255] Table 1 lists 29 AhR ligands used in this disclosure as potential biomarkers for predicting a patient’s cancer response to ICI therapy.

[0256] Table 1

[0257]

[0258]

[0259] Example 1 shows that among the 29 AhR ligands, kynurenine, tryptophan, phenylpyruvate, and indolelactate were differentially secreted in fecal samples at baseline, with an FDR-corrected p-value of less than 0.05 when comparing patients who became responsive to ICI therapy (responders) with patients who did not respond to any ICI therapy (non-responders). All four AhR ligands were elevated in fecal samples from non-responders compared to responders. Furthermore, no statistically significant differences were found in plasma samples from non-responders compared to responders for these four AhR ligands.

[0260] In some embodiments, the biomarkers according to this disclosure comprise one or more of kynurenine, tryptophan, phenylpyruvic acid, and indolelacic acid. For example, in some embodiments, the biomarkers according to this disclosure comprise kynurenine. In some embodiments, the biomarkers according to this disclosure comprise tryptophan. In some embodiments, the biomarkers according to this disclosure comprise phenylpyruvic acid. In some embodiments, the biomarkers according to this disclosure comprise indolelacic acid.

[0261] In some embodiments, the biomarkers according to this disclosure comprise kynurenine and tryptophan. In some embodiments, the biomarkers according to this disclosure comprise kynurenine and phenylpyruvate. In some embodiments, the biomarkers according to this disclosure comprise kynurenine and indolelacic acid. In some embodiments, the biomarkers according to this disclosure comprise tryptophan and phenylpyruvate. In some embodiments, the biomarkers according to this disclosure comprise tryptophan and indolelacic acid. In some embodiments, the biomarkers according to this disclosure comprise phenylpyruvate and indolelacic acid.

[0262] In some embodiments, the biomarkers according to this disclosure comprise kynurenine, tryptophan, and phenylpyruvate. In some embodiments, the biomarkers according to this disclosure comprise kynurenine, tryptophan, and indolelactate. In some embodiments, the biomarkers according to this disclosure comprise tryptophan, phenylpyruvate, and indolelactate.

[0263] In some embodiments, the biomarkers according to this disclosure include kynurenine, tryptophan, phenylpyruvic acid, and indolelacic acid.

[0264] This disclosure uses biomarkers to predict a patient's cancer response to ICI therapy. Several ICI therapies have been approved by the Food and Drug Administration for the treatment of, in particular, melanoma, non-small cell lung cancer, renal cell carcinoma, head and neck squamous cell carcinoma, Hodgkin's lymphoma, urothelial carcinoma, small cell lung cancer, esophageal squamous cell carcinoma, cervical cancer, primary mediastinal large B-cell lymphoma, MSI-H / dMMR colorectal cancer, hepatocellular carcinoma, Merkel cell carcinoma, triple-negative breast cancer, and cutaneous squamous cell carcinoma.

[0265] In some embodiments, ICI therapy includes inhibiting one or more of the following: CTLA-4, PD-1, PD-L1, VISTA, B7-H2, B7-H3, B7-H4, B7-H6, ICOS, HVEM, PD-L2, CD160, gp49B, PIR-B, KIR family receptors, TIM-1, TIM-3, TIM-4, LAG-3, GITR, 4-IBB, OX-40, BTLA, SIRP, CD47, CD48, 2B4 (CD244), B7.1, B7.2, ILT-2, ILT-4, TIGIT, HHLA2, prolactin, IDO, CD39, CD73, and A2aR.

[0266] CTLA-4 and PD-1 are negative regulators of T-cell immune function and play a role at different stages of the immune response. Although CTLA-4 regulates the early activation of naive and memory T cells, typically within lymph nodes, the PD-1 pathway regulates the response of previously activated T cells to inflammatory signals in the later stages of the immune response, primarily in peripheral tissues and within the cancer itself (Fife et al., Immunol Rev., 2008, 224:166-182).

[0267] PD-1 is highly expressed by activated T cells, B cells, dendritic cells (DCs), and natural killer (NK) cells, while PD-L1 is expressed on several types of tumor cells. PD-1 is regulated by two ligands, programmed death-ligand 1 (PD-L1; B7-H1, CD274) and PD-L2 (B7-DC), leading to inhibition of T cell activation. PD-1 binding inhibits T cell proliferation and the production of pro-inflammatory cytokines such as interferon-γ (IFN-γ), tumor necrosis factor-α, and IL-2 (Chamoto et al., Curr Top Microbiol Immunol. [Recent Issues in Microbiology and Immunology], 2017, 410:75-97). The binding of PD-L1 and PD-1 in normal tissues maintains immune system homeostasis and protects against autoimmunity during infection or inflammation. Their interaction in the tumor microenvironment appears to block these interactions by shutting down cytotoxic T cells, which provide tumor cells with an immune escape mechanism.

[0268] CTLA-4, PD-1, and PD-L1 are well-validated targets for ICI therapy. Approved ICI therapies targeting these checkpoint proteins include pembrolizumab (Keytruda), nivolumab (Opdivo), ipilimumab (Yervoy), acitumab (Bavencio), atezolizumab (Tecentriq), durvalumab (Imfinzi), cimiprimab (LBTAYO), sintilimab (Tyvyt), toripalimab (Tuoyi), and camrelizumab (AiRuiKa).

[0269] Ongoing clinical trials include therapeutics targeting LAG-3, TIM-3TIGIT, and VISTA (Qin et al., Molecular Cancer, 2019, 18:155). Non-limiting examples of ICI therapies targeting LAG-3 include IMP321 (eftilagimod alpha), renalalimab (BMS-986016), LAG525, MK-4280, REGN3767, TSR-033, BI754111, Sym022, FS118, and MGD013. Non-limiting examples of ICI therapies targeting TIM-3 include TSR-022, MBG453, Sym023, INCAGN2390, LY3321367, BMS-986258, SHR-1702, and RO7121661. Non-limiting examples of ICI therapies targeting TIGIT include MK-7684, etigilimab (OMP-313), tiragolumab (MTIG7192A, RG-6058), BMS-986207, AB-154, and ASP-8374. Non-limiting examples of ICI therapies targeting VISTA include JNJ-61610588 and CA-170.

[0270] In some embodiments, the ICI therapy according to this disclosure includes anti-PD-1 antibody, anti-PD-L1 antibody, anti-CTLA-4 antibody, anti-LAG-3 antibody, anti-TIM-3 antibody, anti-TIGIT antibody, anti-VISTA antibody, or a combination thereof.

[0271] In some embodiments, the ICI therapies disclosed herein include pembrolizumab (Keytruda), nivolumab (Opdivo), ipilimumab (Yervoy), acitumab (Bavencio), atezolizumab (Tecentriq), durvalumab (Imfinzi), cimiprimab (LBTAYO), sintilimab (Tyvyt), toripalimab (Tuoyi), camrelizumab (AiRuiKa), and IMP321 (Eftilagimod). Alpha), piracetamab (BMS-986016), LAG525, MK-4280, REGN3767, TSR-033, BI754111, Sym022, FS118, MGD013, TSR-022, MBG453, Sym023, INCAGN2390, LY3321367, BMS-986258, SHR-1702, RO7121661, MK-7684, ettiribumab (OMP-313), tislelizumab (MTIG7192A, RG-6058), BMS-986207, AB-154, ASP-8374, JNJ-61610588, CA-170, or combinations thereof.

[0272] According to this disclosure, the amount of biomarkers in a biological sample is measured using one or more methods known in the art. In some embodiments, the biological sample includes a plasma sample taken from a cancer patient. In some embodiments, the biological sample includes a serum, plasma, or fecal sample (e.g., an intestinal lumen sample) taken from a cancer patient.

[0273] Diagnostic / prognostic and therapeutic uses of biomarkers

[0274] The biomarkers disclosed herein can be used in a variety of diagnostic / prognostic and therapeutic applications, or combinations thereof.

[0275] The biomarkers disclosed herein can be used in in vitro methods to predict the responsiveness of a patient's cancer to ICI therapy. In some embodiments, the method includes (a) measuring or having measured the amount of at least one biomarker from a serum, plasma, or fecal sample obtained from the patient, (b) comparing or having compared the amount of the at least one biomarker to a threshold fraction, and (c) determining that the patient's cancer may be responsive to ICI therapy if the amount of the at least one biomarker is equal to or less than the threshold fraction.

[0276] The biomarkers disclosed herein can be used in an in vitro method to determine whether a patient will benefit from treatment with an AhR antagonist. In some embodiments, the method includes (a) measuring or having measured the amount of at least one biomarker from a serum, plasma, or fecal sample obtained from the patient, (b) comparing or having compared the amount of the at least one biomarker to a threshold fraction, and (c) determining that the patient will benefit from treatment with an AhR antagonist if the amount of the at least one biomarker is above the threshold fraction.

[0277] The biomarkers disclosed herein can be used to stratify cancer patients based on the amount of at least one biomarker present in serum, plasma, or fecal samples. In some embodiments, such stratification can be used to inform and provide effective treatment options for cancer patients.

[0278] In some embodiments, a patient may respond to ICI therapy even without combining ICI therapy with an AhR antagonist when the amount of at least one biomarker in a patient's serum, plasma, or stool sample is equal to or below a threshold fraction of that biomarker. In such embodiments, the patient can be treated with the ICI therapy disclosed herein. Therefore, in some embodiments, this disclosure provides a method of treating cancer in a patient in need, comprising: (a) measuring or having measured the amount of at least one biomarker obtained from a patient's serum, plasma, or stool sample; (b) comparing or having compared the amount of the at least one AhR ligand with a threshold fraction; and (c) treating the patient with ICI therapy if the amount of the at least one biomarker is equal to or below a threshold fraction. In some embodiments, ICI therapy may optionally be combined with various cancer therapies known in the art, such as surgery, chemotherapy, or radiation therapy to remove cancerous tissue. In some embodiments, the various cancer therapies are not AhR antagonists. In some embodiments, the various cancer therapies are AhR antagonists. In some embodiments, the at least one AhR ligand is selected from piperine, nicotinamide, 4-hydroxyphenylacetic acid, urolithin A, berberine, quinolinic acid, bilirubin, phenyl acetate, citrulline, phenylpyruvic acid, xanthuric acid, spermidine, indolepropionate, pyridoxine (vitamin B6), indole, indolelactic acid, kynurenic acid, curcumin, biliverdin, indoleacetic acid, pyridinecarboxylate, kynurenine, tryptophan, 3-indophenol sulfate, serotonin, phenyl lactate (PLA), tryptophan, benzoate, and 4-hydroxyphenylpyruvic acid. In some embodiments, the at least one AhR ligand comprises two or more of the following: piperine, nicotinamide, 4-hydroxyphenylacetate, urolithin A, berberine, quinolinic acid, bilirubin, phenyl acetate, citrulline, phenylpyruvic acid, xanthuric acid, spermidine, indolepropionate, pyridoxine (vitamin B6), indole, indolelactic acid, kynurenic acid, curcumin, biliverdin, indoleacetate, pyridinecarboxylate, kynurenine, tryptophan, 3-indophenol sulfate, serotonin, phenyl lactate (PLA), tryptophan, benzoate, and 4-hydroxyphenylpyruvic acid. In some embodiments, the at least one AhR ligand comprises three or more of the following: piperine, nicotinamide, 4-hydroxyphenylacetic acid, urolithin A, berberine, quinolinic acid, bilirubin, phenylacetic acid, citrulline, phenylpyruvic acid, xanthuric acid, spermidine, indolepropionate, pyridoxine (vitamin B6), indole, indolelactic acid, kynurenic acid, curcumin, biliverdin, indoleacetic acid, pyridinecarboxylate, kynurenine, tryptophan, 3-indophenol sulfate, serotonin, phenyl lactate (PLA), tryptophan, benzoate, and 4-hydroxyphenylpyruvic acid ester. In some embodiments, the at least one AhR ligand comprises kynurenine, tryptophan, phenylpyruvic acid, indolelactic acid, or a combination thereof.

[0279] In some embodiments, when the amount of the at least one biomarker in a patient's serum, plasma, or stool sample is higher than a threshold fraction of the biomarker, the patient may be unresponsive to ICI therapy in the absence of an AhR antagonist (e.g., ICI therapy alone or a combination of ICI therapy with different (non-AhR antagonist) cancer therapies, such as surgery, chemotherapy, and / or radiation therapy). In such embodiments, the patient may be treated with a combination of ICI therapy and an AhR antagonist. Therefore, in some embodiments, this disclosure provides a method of treating cancer in a patient in need, comprising: (a) measuring or having measured the amount of at least one biomarker obtained from a patient's serum, plasma, or stool sample; (b) comparing or having compared the amount of the at least one AhR ligand with a threshold fraction; (c) if the amount of the at least one biomarker is higher than a threshold fraction, determining or having determined that the patient will benefit from treatment with an AhR antagonist; and (d) treating the patient determined to benefit in (d) with an effective amount of the AhR antagonist.

[0280] In some embodiments, this disclosure provides a method of treating cancer in a patient in need with a combination therapy comprising an ICI therapy and an AhR antagonist, comprising: (a) measuring or having measured the amount of at least one biomarker from a serum, plasma, or fecal sample obtained from the patient; (b) comparing or having compared the amount of the at least one biomarker to a threshold fraction; (c) if the amount of the at least one biomarker is above the threshold fraction, determining or having determined that the patient will benefit from the combination therapy; and (d) treating the patient determined to benefit in (c) with the combination therapy.

[0281] In some embodiments of the diagnostic / prognostic and treatment methods disclosed herein, the biomarkers are selected from piperine, nicotinamide, 4-hydroxyphenylacetate, urolithiasis A, berberine, quinolinic acid, bilirubin, phenyl acetate, citrulline, phenylpyruvic acid, xanthuric acid, spermidine, indolepropionate, pyridoxine (vitamin B6), indole, indolelactic acid, kynurenic acid, curcumin, biliverdin, indoleacetate, pyridinecarboxylate, kynurenine, tryptophan, 3-indophenol sulfate, serotonin, phenyl lactate (PLA), tryptophan, benzoate, and 4-hydroxyphenylpyruvic acid. For example, in some embodiments, the biomarker comprises two or more of the following: piperine, nicotinamide, 4-hydroxyphenylacetate, urolithin A, berberine, quinolinic acid, bilirubin, phenyl acetate, citrulline, phenylpyruvic acid, xanthuric acid, spermidine, indolepropionate, pyridoxine (vitamin B6), indole, indolelactic acid, kynurenic acid, curcumin, biliverdin, indoleacetate, pyridinecarboxylate, kynurenine, tryptophan, 3-indophenol sulfate, serotonin, phenyl lactate (PLA), tryptophan, benzoate, and 4-hydroxyphenylpyruvic acid. In some embodiments, the biomarker comprises three or more of the following: piperine, nicotinamide, 4-hydroxyphenylacetate, urolithin A, berberine, quinolinic acid, bilirubin, phenyl acetate, citrulline, phenylpyruvic acid, xanthuric acid, spermidine, indolepropionate, pyridoxine (vitamin B6), indole, indolelactic acid, kynurenic acid, curcumin, biliverdin, indoleacetate, pyridinecarboxylate, kynurenine, tryptophan, 3-indophenol sulfate, serotonin, phenyl lactate (PLA), tryptophan, benzoate, and 4-hydroxyphenylpyruvic acid.

[0282] In some embodiments of the diagnostic / prognostic and treatment methods disclosed herein, the biomarker comprises kynurenine, tryptophan, phenylpyruvate, indolelacic acid, or a combination thereof. For example, in some embodiments, the biomarker comprises kynurenine. In some embodiments, the biomarker comprises tryptophan. In some embodiments, the biomarker comprises phenylpyruvate. In some embodiments, the biomarker comprises indolelacic acid. In some embodiments, the biomarker comprises kynurenine and tryptophan. In some embodiments, the biomarker comprises kynurenine and phenylpyruvate. In some embodiments, the biomarker comprises kynurenine and indolelacic acid. In some embodiments, the biomarker comprises tryptophan and phenylpyruvate. In some embodiments, the biomarker comprises tryptophan and indolelacic acid. In some embodiments, the biomarker comprises phenylpyruvate and indolelacic acid. In some embodiments, the biomarker comprises kynurenine, tryptophan, and phenylpyruvate. In some embodiments, the biomarker comprises kynurenine, tryptophan, and indolelacic acid. In some embodiments, the biomarker comprises tryptophan, phenylpyruvate, and indolelacic acid. In some embodiments, the biomarkers include kynurenine, tryptophan, phenylpyruvic acid, and indolelacic acid.

[0283] In some embodiments of the diagnostic / prognostic and treatment methods disclosed herein, cancer patients with non-small cell lung cancer, small cell lung cancer, head and neck squamous cell carcinoma, renal cell carcinoma, gastric adenocarcinoma, nasopharyngeal carcinoma, urothelial carcinoma, colorectal cancer, pleural mesothelioma, triple-negative breast cancer, esophageal tumors, multiple myeloma, gastric and gastroesophageal junction cancer, melanoma, Hodgkin lymphoma, hepatocellular carcinoma, lung cancer, head and neck cancer, non-Hodgkin lymphoma, metastatic clear cell renal cell carcinoma, lung squamous cell carcinoma, mesothelioma, gastric cancer, gastroesophageal junction cancer, metastatic melanoma, and metastatic non-cutaneous melanoma are included. Melanoma, urothelial carcinoma, diffuse large B-cell lymphoma, renal cell carcinoma, ovarian cancer, fallopian tube cancer, peritoneal tumor, extensive-stage small cell lung cancer, bladder cancer, transitional cell carcinoma, prostate tumor, recurrent or metastatic PD-L1 positive or negative squamous cell carcinoma of the head and neck, recurrent squamous cell lung cancer, advanced solid malignant tumors, pharyngeal squamous cell carcinoma, laryngeal squamous cell carcinoma, unresectable or metastatic melanoma, biliary tract tumors, esophageal squamous cell carcinoma, breast cancer, pancreatic cancer, glioblastoma, metastatic cancer, prostate cancer, solid organ cancer, gastric cancer, colon cancer, or liver cancer.

[0284] In some embodiments of the diagnostic / prognostic and treatment methods disclosed herein, cancer patients have previously been treated with ICI therapy but have not responded to it. In some embodiments, cancer patients have previously been treated with ICI therapy and have developed resistance to it. In some embodiments, ICI therapy includes anti-PD-1 antibody, anti-PD-L1 antibody, anti-CTLA-4 antibody, anti-LAG-3 antibody, anti-TIM-3 antibody, anti-TIGIT antibody, anti-VISTA antibody, or a combination thereof. In some embodiments, ICI therapy includes pembrolizumab (Keytruda), nivolumab (Opdivo), ipilimumab (Yervoy), acitumab (Bavencio), atezolizumab (Tecentriq), durvalumab (Imfinzi), cimiprimab (LBTAYO), sintilimab (Tyvyt), toripalimab (Tuoyi), camrelizumab (AiRuiKa), and IMP321 (Eftilagimod). Alpha), piracetamab (BMS-986016), LAG525, MK-4280, REGN3767, TSR-033, BI754111, Sym022, FS118, MGD013, TSR-022, MBG453, Sym023, INCAGN2390, LY3321367, BMS-986258, SHR-1702, RO7121661, MK-7684, ettiribumab (OMP-313), tislelizumab (MTIG7192A, RG-6058), BMS-986207, AB-154, ASP-8374, JNJ-61610588, CA-170, or combinations thereof.

[0285] In some embodiments of the diagnostic / prognostic and treatment methods disclosed herein, cancer patients may have been previously treated or are currently being treated with different (non-ICI and non-AhR antagonist) cancer therapies, such as surgical removal of cancerous tissue, chemotherapy, or radiation therapy.

[0286] In some embodiments of the diagnostic / prognostic and treatment methods disclosed herein, the amounts of the biomarkers disclosed herein are measured at appropriate times in serum, plasma, or stool samples taken from cancer patients. In some embodiments, samples are collected approximately one week after the start of ICI therapy. In some embodiments, samples are collected approximately two weeks after the start of ICI therapy. In some embodiments, samples are collected approximately three weeks after the start of ICI therapy. In some embodiments, samples are collected approximately four weeks after the start of ICI therapy. In some embodiments, samples are collected approximately five weeks after the start of ICI therapy. In some embodiments, samples are collected approximately six weeks after the start of ICI therapy. In some embodiments, samples are collected approximately seven weeks after the start of ICI therapy. In some embodiments, samples are collected approximately eight weeks after the start of ICI therapy. In some embodiments, samples are collected approximately nine weeks after the start of ICI therapy. In some embodiments, samples are collected approximately ten weeks after the start of ICI therapy. In some embodiments, samples are collected more than ten weeks, three months, five months, seven months, ten months, one year, two years, three years, four years, or five years after the start of ICI therapy.

[0287] Sample preparation can involve any procedure known in the art. Such procedures include, by way of example only, concentration, dilution, pH adjustment, removal of highly abundant peptides (e.g., albumin, gamma globulin, and transferrin), addition of preservatives and calibrators, addition of protease inhibitors, addition of denaturing agents, sample desalting, sample protein concentration, and extraction and purification of lipids.

[0288] After sample preparation, the amount of biomarker in serum, plasma, or fecal samples is measured using one or more applicable methods known in the art. Suitable measurement / detection methods may include mass spectrometry, such as liquid chromatography-mass spectrometry (LC-MS), gas chromatography-mass spectrometry (GC-MS), or tandem mass spectrometry (MS-MS). Other suitable methods include reversed-phase chromatography (e.g., with positive and / or negative ion modes) and hydrophobic interactive liquid ion chromatography (HILIC) (e.g., with positive and / or negative ion modes), or combinations thereof. In some embodiments, the amount of biomarker is measured by LC-MS. In some embodiments, the amount of biomarker is analyzed using ultra-high performance liquid chromatography-tandem mass spectrometry. See Example 1.

[0289] In some embodiments, fecal samples may be graded prior to the application of measurement / detection methods. In some embodiments, the amount of a biomarker may be measured by methods that do not require physical separation of the biomarker itself. For example, nuclear magnetic resonance (NMR) spectroscopy may be used to resolve the amount of a biomarker from a complex mixture of metabolites including various AhR ligands.

[0290] In some embodiments of the diagnostic / prognostic and treatment methods disclosed herein, the amount of each biomarker is compared to a predetermined threshold score for that biomarker. In some embodiments, the threshold score is the amount of the biomarker from serum, plasma, or stool samples of control cancer patients responding to ICI therapy. In some embodiments, the threshold score may be generated from a population or cohort of two or more control cancer patients responding to ICI therapy. A population or cohort may, for example, include at least 2, 3, 4, 5, 10, 15, 18, 20, 30, 40, 50, 75, 100, or more cancer patients. In such embodiments, the threshold score for each biomarker may be the average amount of the biomarker from serum, plasma, or stool samples of a population or cohort of control cancer patients responding to ICI therapy.

[0291] To determine the threshold fraction for each biomarker, serum, plasma, or stool samples may be obtained from control cancer patients who responded to ICI therapy at an appropriate time, such as those disclosed herein. Control cancer patients may respond to ICI therapy including inhibition of one or more of the following: CTLA-4, PD-1, PD-L1, VISTA, B7-H2, B7-H3, B7-H4, B7-H6, ICOS, HVEM, PD-L2, CD160, gp49B, PIR-B, KIR family receptors, TIM-1, TIM-3, TIM-4, LAG-3, GITR, 4-IBB, OX-40, BTLA, SIRP, CD47, CD48, 2B4 (CD244), B7.1, B7.2, ILT-2, ILT-4, TIGIT, HHLA2, lactolipoprotein, IDO, CD39, CD73, and A2aR. In some embodiments, ICI therapy includes anti-PD-1 antibody, anti-PD-L1 antibody, anti-CTLA-4 antibody, anti-LAG-3 antibody, anti-TIM-3 antibody, anti-TIGIT antibody, anti-VISTA antibody, or a combination thereof. In some embodiments, ICI therapy includes pembrolizumab (Keytruda), nivolumab (Opdivo), ipilimumab (Yervoy), acitumab (Bavencio), atezolizumab (Tecentriq), durvalumab (Imfinzi), cimiprimab (LBTAYO), sintilimab (Tyvyt), toripalimab (Tuoyi), camrelizumab (AiRuiKa), and IMP321 (Eftilagimod). Alpha), piracetamab (BMS-986016), LAG525, MK-4280, REGN3767, TSR-033, BI754111, Sym022, FS118, MGD013, TSR-022, MBG453, Sym023, INCAGN2390, LY3321367, BMS-986258, SHR-1702, RO7121661, MK-7684, ettiribumab (OMP-313), tislelizumab (MTIG7192A, RG-6058), BMS-986207, AB-154, ASP-8374, JNJ-61610588, CA-170, or combinations thereof.

[0292] The amount of a biomarker in a serum, plasma, or stool sample from a cancer patient may be “higher” than a threshold fraction of the biomarker, for example, at least 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, 300%, 350%, 400%, 500%, 600%, 700%, 800%, 900%, 1000%, or more above the threshold fraction. In some embodiments, the amount of a biomarker in a serum, plasma, or stool sample from a cancer patient being tested may be “lower” than a threshold fraction of the biomarker, for example, at least about 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more below the threshold fraction.

[0293] In some embodiments of the treatment methods disclosed herein, an AhR antagonist may be administered in combination with ICI therapy to a cancer patient when the level of a biomarker in a serum, plasma, or stool sample exceeds a threshold fraction. In these embodiments, the AhR antagonist may be administered concurrently with ICI therapy. In some embodiments, the AhR antagonist and ICI therapy are administered sequentially. In some embodiments, the combination therapy results in an increased or synergistic antitumor response compared to treatment with the respective ICI therapy alone.

[0294] Pyridopyrimidine derivatives as AhR antagonists

[0295] This article discloses novel 6,8-trisubstituted pyrido[3,4-d]pyrimidine-4(3H)-one compounds of formula (I), (Ia), or (Ib), or pharmaceutically acceptable salts thereof, which effectively inhibit AhR and are therefore, as known AhR antagonists, available for use in the therapeutic methods disclosed herein.

[0296] Unless otherwise specified, the nomenclature used to describe chemical groups or parts as used herein follows the convention of reading the name from left to right, with the attachment point to the rest of the molecule on the right side of the name. For example, the group "(C 1-3 Alkoxy)C 1-3 The "alkyl" terminus is attached to the rest of the molecule at the alkyl terminus. Further examples include methoxyethyl, where the attachment point is at the ethyl terminus, and methylamino, where the attachment point is at the amine terminus.

[0297] Unless otherwise specified, when a chemical group is described by its chemical formula or a structure having a terminal bonded portion indicated by "-", it should be understood that "-" represents an attachment point. In some embodiments, the wavy line (i.e., Describe the attachment points.

[0298] As used herein, the term "pharmaceutically acceptable salt" means a salt that is pharmaceutically acceptable as defined herein and has the desired pharmacological activity of the parent compound. Non-limiting examples of pharmaceutically acceptable salts include those derived from inorganic acids, including hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, and phosphoric acid; and those derived from organic acids, including acetic acid, trifluoroacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, stearic acid, malic acid, maleic acid, malonic acid, salicylic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, p-toluenesulfonic acid, methanesulfonic acid, ethanesulfonic acid, and lactic acid. Additional non-limiting examples of pharmaceutically acceptable salts include those formed when an acidic proton in the parent compound is replaced by a metal ion. Non-limiting examples of metal ions include alkali metal ions and alkaline earth metal ions, as well as those formed when an acidic proton present in the parent compound is replaced by an ammonium ion, primary ammonium ion, secondary ammonium ion, tertiary ammonium ion, or quaternary ammonium ion. Non-limiting examples of alkali metals and alkaline earth metals include sodium, potassium, lithium, calcium, aluminum, magnesium, copper, zinc, iron, and manganese. Additional non-limiting examples of pharmaceutically acceptable salts include those containing one or more counterions and zwitterions.

[0299] As used herein, “acyl” or “alkanoyl” is a functional group having the formula RCO-, where R is bonded to a carbon atom of a carbonyl functional group via a single bond, and “-” indicates an attachment point to the rest of the molecule. Non-limiting examples of acyl groups include formyl (HC(O)-, also known as methanoyl)), acetyl (CH3C(O)-, also known as ethanoyl) and benzoyl (PhC(O)-).

[0300] As used herein, the terms "alkyl" or "aliphatic" refer to a fully saturated, straight (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain having a single attachment point to the rest of the molecule. Unless otherwise stated, an alkyl group is a hydrocarbon chain having 1 to 20 alkyl carbon atoms. In some embodiments, the alkyl group comprises one to twelve carbon atoms (C1-C2). 12In some embodiments, the alkyl group comprises one to eight carbon atoms (C1-C8). In some embodiments, the alkyl group comprises one to six carbon atoms (C1-C6). In some embodiments, the alkyl group comprises one to four carbon atoms (C1-C4). In some embodiments, the cyclic alkyl group comprises three to six carbon atoms (C3-C6). Non-limiting examples of substituted and unsubstituted straight-chain, branched, and cyclic alkyl groups include methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, cyclobutyl, cyclopentyl, cyclohexyl, hydroxymethyl, chloromethyl, fluoromethyl, trifluoromethyl, aminomethyl, 2-aminoethyl, 3-aminopropyl, 4-aminobutyl, dimethylaminomethyl, 2-dimethylaminoethyl, 3-dimethylaminopropyl, 4-dimethylaminobutyl, cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl, trifluoroethyl, and trifluoropropyl.

[0301] As used herein, “alkoxy” refers to an alkyl group, as previously defined, attached to the main carbon chain via an oxygen (“alkoxy”) atom.

[0302] As used herein, “halogenated” and “halogen” are interchangeable and refer to halogen atoms such as fluorine (F), chlorine (Cl), bromine (Br) and iodine (I).

[0303] "Halogenated alkyl" refers to an alkyl group that is substituted by one or more halogen atoms (F, Cl, Br, I). For example, "fluoromethyl" refers to a methyl group that is substituted by one or more fluorine atoms (e.g., monofluoromethyl, difluoromethyl, or trifluoromethyl).

[0304] "Haloalkoxy" refers to an alkoxy group that is substituted by one or more halogen atoms (F, Cl, Br, I). For example, "fluoromethoxy" refers to a methoxy group that is substituted by one or more fluorine atoms (e.g., monofluoromethoxy, difluoromethoxy, or trifluoromethoxy).

[0305] "Hydroxyalkyl" refers to an alkyl group that is replaced by one or more hydroxyl groups (-OH).

[0306] As used interchangeably herein, the terms "cycloalkyl" and "cycloalkyl group" refer to a cyclic, saturated, monovalent hydrocarbon group having three to twelve carbon atoms and a single attachment point to the remainder of the molecule. The cycloalkyl group may be unsubstituted or substituted. In some embodiments, the cycloalkyl group comprises three to eight carbon atoms (C3-C8). In some embodiments, the cycloalkyl group comprises three to six carbon atoms (C3-C6). Non-limiting examples of substituted and unsubstituted cycloalkyl groups include cyclopropyl, cyclopropylmethyl, cyclobutyl, cyclobutylmethyl, cyclopentyl, cyclopentylmethyl, cyclohexyl, cycloheptyl, and cyclooctyl.

[0307] As used interchangeably in this article, the terms "alkylene" and "alkylene group" refer to a group containing one to twelve carbon atoms (C1-C2). 12 The alkylene group is a saturated divalent (i.e., having two attachment sites with the rest of the molecule) hydrocarbon group. The alkylene group can be straight-chain, branched, or cyclic. The alkylene group can be unsubstituted or substituted. In some embodiments, the alkylene group comprises one to eight carbon atoms (C1-C8). In some embodiments, the alkylene group comprises one to six carbon atoms (C1-C6). In some embodiments, the alkylene group comprises one to four carbon atoms (C1-C4). Non-limiting examples of alkylene groups include methylene and ethylene.

[0308] As used interchangeably herein, the terms "alkenyl" and "alkenyl group" refer to a monovalent (i.e., having a single attachment point with the rest of the molecule) hydrocarbon group comprising two to eight carbon atoms (C2-C8) and having at least one unsaturated site (i.e., sp2 carbon-carbon double bond). Alkenyl groups can be linear, branched, or cyclic. Alkenyl groups can be unsubstituted or substituted. In some embodiments, an alkenyl group comprises two to six carbon atoms (C2-C6). In some embodiments, an alkenyl group comprises two to four carbon atoms (C2-C4). Alkenyl groups can have an E or Z orientation. Non-limiting examples of alkenyl groups include vinyl (also known as vinyl), 1-propenyl, isopropenyl, and 2-chlorovinyl.

[0309] As used interchangeably herein, the terms "alkenyl" and "alkenyl group" refer to a divalent (i.e., having two attachment sites with the rest of the molecule) hydrocarbon group having at least one unsaturated site (e.g., an sp2 carbon-carbon double bond). Alkenyl groups can be linear, branched, or cyclic. Alkenyl groups can be unsubstituted or substituted. In some embodiments, an alkylene group comprises two to six carbon atoms (C2-C6). In some embodiments, an alkylene group comprises two to four carbon atoms (C2-C4). Alkenyl groups can have an E or Z orientation. A non-limiting example of an alkenyl group is vinylene (also known as vinylene).

[0310] As used interchangeably herein, the terms "alkynyl" and "alkynyl group" refer to a monovalent (i.e., having a single attachment point to the rest of the molecule) hydrocarbon group having at least one unsaturated site (i.e., an sp carbon-carbon triple bond) and containing two to eight carbon atoms (C2-C8). The alkynyl group can be straight-chain or branched. The alkynyl group can be unsubstituted or substituted. In some embodiments, the alkynyl group comprises two to six carbon atoms (C2-C6). In some embodiments, the alkynyl group comprises two to four carbon atoms (C2-C4). A non-limiting example of an alkynyl group is the ethynyl group.

[0311] As used interchangeably herein, the terms "ynynyl" and "ynynyl group" refer to a divalent (i.e., having two attachment sites with the rest of the molecule) hydrocarbon group having at least one unsaturated site (i.e., an sp carbon-carbon triple bond) and containing two to eight carbon atoms (C2-C8). The ynynyl group can be straight-chain or branched. The ynynyl group can be unsubstituted or substituted. In some embodiments, the ynynyl group comprises two to six carbon atoms (C2-C6). In some embodiments, the ynynyl group comprises two to four carbon atoms (C2-C4). A non-limiting example of an ynynyl group is ethynylene.

[0312] As used herein, "aromatic group" or "aromatic ring" refers to a chemical group containing a conjugated planar ring system having delocalized π-electron orbitals consisting of [4n+2]p orbital electrons, where n is an integer ranging from 0 to 6. Non-limiting examples of aromatic groups include aryl and heteroaryl groups.

[0313] As used interchangeably in this article, the terms "aryl" and "aryl group" refer to 6-20 carbon atoms (C6-C6). 20 The aryl group is a single aryl group (i.e., having a single attachment point with the rest of the molecule). The aryl group can be unsubstituted or substituted. Non-limiting examples of unsubstituted and substituted aryl groups include phenyl, 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 2-methylphenyl, 3-methylphenyl, 4-methylphenyl, 2-chlorophenyl, 3-chlorophenyl, 4-chlorophenyl, 2,6-dichlorophenyl, 3,4-difluorophenyl, 2-hydroxyphenyl, 3-hydroxyphenyl, 4-hydroxyphenyl, 2-methoxyphenyl, 3-methoxyphenyl, 4-methoxyphenyl, 2-phenoxyphenyl, 3-phenoxyphenyl, 4-phenoxyphenyl. Phenyl, 2-cyanophenyl, 3-cyanophenyl, 4-cyanophenyl, 2-dimethylaminophenyl, 3-dimethylaminophenyl, 4-dimethylaminophenyl, 3-methylsulfonylphenyl, 4-methylsulfonylphenyl, 3-aminophenyl, 3-methylaminophenyl, 3-(2-hydroxyethoxy)phenyl, 2-trifluoromethylphenyl, 3-trifluoromethylphenyl, 4-trifluoromethylphenyl, 2-isopropylphenyl, 3-isopropylphenyl, 4-isopropylphenyl, 1-naphthyl and 2-naphthyl.

[0314] As used herein, the term "heteroalkyl" refers to an alkyl group in which at least one carbon atom in the chain is replaced by a heteroatom such as nitrogen, oxygen, phosphorus, or sulfur. Heteroalkyl groups can be unsubstituted or substituted.

[0315] As used interchangeably herein, the terms “heterocyclic alkyl,” “heterocyclic,” “heterocyclic group,” and “heterocyclic alkyl group” refer to a saturated or partially unsaturated ring system having 3 to 20 atoms, wherein at least one of these ring atoms is a heteroatom, such as nitrogen, oxygen, phosphorus, and sulfur. Heterocyclic alkyl groups may be unsubstituted or substituted. In some embodiments, a heterocyclic alkyl group comprises 3 to 10 atoms. In some embodiments, a heterocyclic alkyl group comprises 3 to 7 atoms. In some embodiments, a heterocyclic alkyl group is monocyclic. In some embodiments, a heterocyclic alkyl group is bicyclic. In some embodiments, a heterocyclic alkyl group comprises a fused ring. Non-limiting examples of unsubstituted and substituted heterocyclic alkyl groups include pyrrolidinyl, N-methylpyrrolidinyl, azaheterocyclic butyl, dihydrofuranyl, tetrahydrofuranyl, tetrahydropyranyl, 3-hydroxypyrrolidinyl, 3-methoxypyrrolidinyl, and benzodioxanepentenyl.

[0316] As used interchangeably herein, the terms "heteroaryl" and "heteroaryl group" refer to an aromatic ring system having 3 to 20 atoms, wherein at least one of these ring atoms is a heteroatom, such as nitrogen, oxygen, phosphorus, and sulfur. Heteroaryl groups can be unsubstituted or substituted. In some embodiments, a heteroaryl group contains 5 to 20 atoms. In some embodiments, a heteroaryl group contains 5 to 9 atoms. In some embodiments, a heteroaryl group contains 5 atoms. In some embodiments, a heteroaryl group contains 6 atoms. In some embodiments, a heteroaryl group contains 7 atoms. In some embodiments, a heteroaryl group is monocyclic. In some embodiments, a heteroaryl group is bicyclic. In some embodiments, a heteroaryl group contains a fused ring. Non-limiting examples of heteroaryl groups include pyridinyl, imidazolyl, imidazopyridinyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furanyl, 2-thiopheneyl, 3-thiopheneyl, isoxazolyl, thiazolyl, oxadiazolyl, 3-methyl-1,2,4-oxadiazolyl, 3-phenyl-1,2,4-oxadiazolyl, oxazolyl, isothiazolyl, pyrroleyl, quinolinyl, isoquinolinyl, tetrahydroisoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, indazoleyl, indoleyl, phthalazinyl, pyridazinyl, triazinyl, thiadiazolyl, furazonyl, benzofuranyl, benzothiopheneyl, benzoxazolyl, quinazolinyl, quinoxolinyl, naphthidyl, furanylpyridinyl, and 1H-pyrrolo[2,3-b]pyridinyl. Non-limiting examples of heteroaryl groups include:

[0317] as well as

[0318] As used herein, the phrase “optionally substituted” means that the specified component may or may not be “substituted”. As used herein, the term “substituted” means that one or more hydrogen atoms on a group (such as an alkyl group, alkylene group, alkenyl group, alkenylene group, alkynyl group, alkenylene group, aryl group, heterocyclic alkyl group, or heteroaryl group) are replaced by one or more substituents. Non-limiting examples of substituents replacing a single hydrogen atom include halogens, hydroxyl groups, and amino groups. Non-limiting examples of substituents replacing two hydrogen atoms include oxo and methylene groups. Non-limiting examples of substituents replacing three hydrogen atoms include nitriles.

[0319] Additional non-limiting examples of substituents include:

[0320] C1-C6 straight-chain, branched, and cyclic alkyl groups, non-limiting examples of which include methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, cyclobutyl, cyclopentyl, and cyclohexyl;

[0321] C2-C8 straight-chain, branched, and cyclic alkenyl groups, non-limiting examples of which include vinyl (also known as vinyl), 1-propenyl, and isopropenyl;

[0322] C2-C8 straight-chain and branched alkynyl groups, non-limiting examples of which include acetylenol;

[0323] Substituted and unsubstituted aryl groups, non-limiting examples of which include phenyl, 2-fluorophenyl, 3-methylphenyl, 4-chlorophenyl, 2,6-dichlorophenyl, 3,4-difluorophenyl, 3-hydroxyphenyl, 4-cyanophenyl, 2-dimethylaminophenyl, 3-methylsulfonylphenyl, 4-trifluoromethylphenyl, 3-isopropylphenyl, 1-naphthyl and 2-naphthyl;

[0324] Substituted and unsubstituted heterocyclic groups, non-limiting examples of which include pyrrolidinyl, N-methylpyrrolidinyl, azahexacyclic butyl, dihydrofuranyl, tetrahydrofuranyl, tetrahydropyranyl, 3-hydroxypyrrolidinyl and 3-methoxypyrrolidinyl;

[0325] Substituted and unsubstituted heteroaryl groups, non-limiting examples of which include pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl, furanyl, 2-thienyl, 3-thienyl, isoxazolyl, thiazolyl, oxadiazolyl, 3-methyl-1,2,4-oxadiazolyl, 3-phenyl-1,2,4-oxadiazolyl, indolyl, benzothiazolyl, and 1H-pyrrolo[2,3-b]pyridinyl;

[0326] -(CR a R b ) z OR cNon-limiting examples include -OH, -OCH3, -OCH2OH and -OCH2CH3;

[0327] -(CR a R b ) z N(R c (R) d Non-limiting examples include -NH2, -NHCH3, -N(CH3)2, -CH2NH2, and -CH2NHCH3.

[0328] Halogen atoms, non-limiting examples of which include fluorine atoms (-F) and chlorine atoms (-Cl);

[0329] -(CR a R b ) z CN;

[0330] -(CR a R b ) z NO2;

[0331] -CH x X y , where X is a halogen atom and the sum of x+y is 3, and non-limiting examples include -CH2F, -CHF2 and -CF3;

[0332] -(CR a R b ) z C(O)R c Non-limiting examples include -COCH3, -COCH2CH3 and -CH2COCH3;

[0333] -(CR a R b ) z C(O)OR c Non-limiting examples include CO2H, -CO2CH3, -CO2CH2CH3, and -CH2CO2CH3.

[0334] -(CR a R b ) z C(O)N(R c (R) d Non-limiting examples of -CONH2, -CONHCH3, -CON(CH3)2, -CH2CONH2, -CH2CONHCH3, and -CH2CON(CH3)2 are included.

[0335] -(CR a R b )z SO2R c Non-limiting examples include -SO2H, -SO2CH3, -CH2SO2H, -CH2SO2CH3, -SO2C6H5, and -CH2SO2C6H5; and

[0336] -(CR a R b ) z SO3R c Non-limiting examples include -SO3H, -SO3CH3, -CH2SO3H, -CH2SO3CH3, -SO3C6H5, and -CH2SO3C6H5;

[0337] Where R a and R b Each of them is independently selected from hydrogen and substituted or unsubstituted C1-C6 straight-chain, branched or cyclic alkyl groups, R c and R d Each of them is independently selected from hydrogen, substituted or unsubstituted C1-C6 straight-chain, branched or cyclic alkyl and aryl groups, or wherein R c and R d Together they form a ring system containing 3 to 7 atoms, and z is selected from 0, 1, 2, 3 and 4.

[0338] In some embodiments, the AhR antagonist according to this disclosure is a compound of formula (I):

[0339]

[0340] Or its pharmaceutically acceptable salt.

[0341] in:

[0342] R 1 and R 2 Each of these is independently selected from optionally substituted alkyl groups, optionally substituted ester groups, optionally substituted heteroalkyl groups, optionally substituted acyl groups, optionally substituted amide groups, optionally substituted aryl groups, optionally substituted heteroaryl groups, optionally substituted cycloalkyl groups, optionally substituted amine groups, and optionally substituted heterocycloalkyl groups; and

[0343] R 3 The group is selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, optionally substituted amide, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted ester, optionally substituted heteroalkyl, optionally substituted heteroaryl, optionally substituted heterocycloalkyl, optionally substituted amine, cyano, halogen, hydroxyl and -C(O)H.

[0344] In some embodiments, R 2 It is a dialkylamine. In some embodiments, R 2It is diethylamine.

[0345] In some embodiments, the AhR antagonist according to this disclosure is a compound of formula Ia:

[0346]

[0347] Or its pharmaceutically acceptable salt.

[0348] in:

[0349] Ring A is selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, and optionally substituted heterocycloalkyl.

[0350] Ring B is selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, and optionally substituted heterocycloalkyl; and

[0351] R is selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, optionally substituted amide, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted ester, optionally substituted heteroalkyl, optionally substituted heteroaryl, optionally substituted heterocycloalkyl, amino, cyano, halogen, hydroxyl and -C(O)H.

[0352] In some embodiments, ring A is selected from 6-10-membered aryl, 5-10-membered heteroaryl, 3-10-membered cycloalkyl, and 3-10-membered heterocycloalkyl, wherein each 6-10-membered aryl, 5-10-membered heteroaryl, 3-10-membered cycloalkyl, and 3-10-membered heterocycloalkyl is independently optionally surrounded by 1 to 5 R A The instance is replaced.

[0353] In some embodiments, ring B is selected from 6-10-membered aryl, 5-10-membered heteroaryl, 3-10-membered cycloalkyl, and 3-10-membered heterocycloalkyl, wherein each 6-10-membered aryl, 5-10-membered heteroaryl, 3-10-membered cycloalkyl, and 3-10-membered heterocycloalkyl is independently optionally surrounded by 1 to 5 R B The instance is replaced.

[0354] In some embodiments, R is selected from hydrogen, C1-C 10 Alkyl, 6-10 aryl, -C(O)R', -C(O)NR'R', 3-10 cycloalkyl, -C(O)OR', C1-C 10 Heteroalkyl, 5-10-membered heteroaryl, 3-10-membered heterocyclic alkyl, amino, cyano, halogenated, hydroxyl, and -C(O)H, wherein each C1-C 10 Alkyl, 6-10 aryl, 3-10 cycloalkyl, C1-C 10 Heteroalkyl, 5-10-membered heteroaryl and 3-10-membered heterocyclic alkyl are independently and optionally bound by 1 to 5 R C The instance is replaced.

[0355] In some embodiments, each R' is independently selected from hydrogen, C1-C2, C2-C3, C4-C4, C5-C6 ... 10 Alkyl, C1-C 10 Haloalkyl, C1-C 10 Hydroxyalkyl and C1-C 10 Heteroalkyl groups.

[0356] In some embodiments, each R A Independently selected from halogenated, hydroxyl-containing, C1-C 10 Alkyl, C1-C 10 Haloalkyl, C1-C 10 Alkoxy, C1-C 10 Halogenated alkoxy groups, C1-C 10 Hydroxyalkyl and NR”R”.

[0357] In some embodiments, each R B Independently selected from halogenated, hydroxyl-containing, C1-C 10 Alkyl, C1-C 10 Haloalkyl, C1-C 10 Alkoxy, C1-C 10 Halogenated alkoxy groups, C1-C 10 Hydroxyalkyl and NR”R”.

[0358] In some embodiments, each R C Independently selected from halogenated, hydroxylated, cyano-based, C1-C 10 Alkyl, C1-C 10 Alkoxy, C1-C 10 Halogenated alkyl, 3-10 membered cycloalkyl, 3-10 membered heterocycloalkyl, 6-10 membered aryl and 5-10 membered heteroaryl.

[0359] In some embodiments, each R” is independently selected from hydrogen, C1-C 10 Alkyl, C1-C 10 Haloalkyl, C1-C 10 Hydroxyalkyl and C1-C 10 Heteroalkyl groups.

[0360] In some embodiments, ring A is selected from 6-10 aryl, 5-8 heteroaryl, 3-10 cycloalkyl, and 3-10 heterocycloalkyl, wherein each 6-10 aryl, 5-10 heteroaryl, 3-10 cycloalkyl, and 3-10 heterocycloalkyl is independently optionally surrounded by 1 to 5 R. A Instance replacement;

[0361] Ring B is selected from 6-10-membered aryl, 5-10-membered heteroaryl, 3-10-membered cycloalkyl, and 3-10-membered heterocycloalkyl, wherein each 6-10-membered aryl, 5-10-membered heteroaryl, 3-10-membered cycloalkyl, and 3-10-membered heterocycloalkyl is independently and optionally surrounded by 1 to 5 R. B Instance replacement;

[0362] R is selected from hydrogen, C1-C 10 Alkyl, 6-10 aryl, -C(O)R', -C(O)NR'R', 3-10 cycloalkyl, -C(O)OR', C1-C10 heteroalkyl, 5-10 heteroaryl, 3-10 heterocycloalkyl, amino, cyano, halogen, hydroxyl, and -C(O)H, wherein each C1-C 10 Alkyl, 6-10 aryl, 3-10 cycloalkyl, C1-C 10 Heteroalkyl, 5-10-membered heteroaryl and 3-10-membered heterocyclic alkyl are independently and optionally bound by 1 to 5 R C Instance replacement;

[0363] Each R' is independently selected from hydrogen, C1-C 10 Alkyl, C1-C 10 Haloalkyl, C1-C 10 Hydroxyalkyl and C1-C 10 Heteroalkyl;

[0364] Each R A Independently selected from halogenated, hydroxyl-containing, C1-C 10 Alkyl, C1-C 10 Haloalkyl, C1-C 10 Alkoxy, C1-C 10 Halogenated alkoxy groups, C1-C 10 Hydroxyalkyl and NR”R”;

[0365] Each R B Independently selected from halogenated, hydroxyl-containing, C1-C 10 Alkyl, C1-C 10 Haloalkyl, C1-C 10 Alkoxy, C1-C 10 Halogenated alkoxy groups, C1-C 10 Hydroxyalkyl and NR”R”;

[0366] Each R C Independently selected from halogenated, hydroxylated, cyano-based, C1-C 10 Alkyl, C1-C 10 Alkoxy, C1-C 10 Halogenated alkyl groups, 3-10 membered cycloalkyl groups, 3-10 membered heterocycloalkyl groups, 6-10 membered aryl groups, and 5-10 membered heteroaryl groups; and

[0367] Each R is independently selected from hydrogen, C1-C 10 Alkyl, C1-C 10 Haloalkyl, C1-C 10 Hydroxyalkyl and C1-C 10 Heteroalkyl groups.

[0368] In some embodiments, ring A is selected from 1 to 5 R's optionally. A Examples of substituted 3-10 membered cycloalkyl groups. In some embodiments, ring A is selected from those optionally replaced by 1 to 5 R groups. A Examples of substituted cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl groups. In some embodiments, ring A is selected from those optionally replaced by 1 to 5 R groups. A The 6-8 aryl groups are replaced by examples. In some embodiments, ring A is optionally replaced by 1 to 3 R groups. A Examples of substituted phenyl groups. In some embodiments, ring A is selected from those optionally replaced by 1 to 5 R groups. A Examples of substituted 5-8 heteroaryl groups.

[0369] In some embodiments, ring A is selected from pyrrolyl, furanyl, furazanyl, thiophene, imidazolyl, isothiazolyl, isoxazolyl, oxazolyl, oxadiazolyl, tetrazolyl, thiazolyl, triazolyl, pyrazole, pyridinyl, pyrazinyl, pyridazinyl, and pyrimidinyl, wherein each of pyrrolyl, furanyl, furazanyl, thiophene, imidazolyl, isothiazolyl, isoxazolyl, oxazolyl, oxadiazolyl, tetrazolyl, thiazolyl, triazolyl, pyrazole, pyridinyl, pyridinyl, pyridazinyl, and pyrimidinyl is independently and optionally selected by 1 to 3 R groups. A The instance is replaced.

[0370] In some embodiments, ring A is optionally surrounded by 1 to 3 R A Examples of substituted pyridinyl groups. In some embodiments, ring A is selected from those optionally replaced by 1 to 5 R groups. A Examples of substituted 5-8 membered heterocyclic alkyl groups. In some embodiments, ring A is selected from pyrrolidinyl, pyrazolyl, piperidinyl, piperazinyl, morpholinyl, and aza-alkyl groups. alkyl, tetrahydropyranyl and tetrahydrofuranyl, wherein pyrrolidinyl, pyrazolylyl, piperidinyl, piperazinyl, morpholinyl, aza Each of the following groups—tetrahydropyranyl and tetrahydrofuranyl—is independently and optionally constituting one to three R groups. A Instances of are replaced. In some embodiments, ring A is optionally replaced by 1 to 3 R A Examples of substituted piperidinyl or morpholinoyl groups.

[0371] In some embodiments, each R A Independently selected from halogenated, C1-C 10 Alkyl, C1-C 10Haloalkyl, C1-C 10 Alkoxy, C1-C 10 Haloalkoxy groups and NR"R". In some embodiments,

[0372] Each R B Independently selected from halogenated, C1-C 10 Alkyl and C1-C 10 Halogenated alkyl groups. In some embodiments, each R C Independently selected from halogenated, hydroxylated, cyano-based, C1-C 10 Alkyl, C1-C 10 Alkoxy, 3-8 membered cycloalkyl, 3-8 membered heterocycloalkyl, and 6-8 membered aryl. In some embodiments, each R” is independently selected from hydrogen and C1-C2. 10 alkyl.

[0373] In some embodiments, each R A Independently selected from halogenated, C1-C 10 Alkyl, C1-C 10 Haloalkyl, C1-C 10 Alkoxy, C1-C 10 Halogenated alkoxy groups and NR"R";

[0374] Each R B Independently selected from halogenated, C1-C 10 Alkyl and C1-C 10 Halogenated alkyl groups;

[0375] Each R C Independently selected from halogenated, hydroxylated, cyano-based, C1-C 10 Alkyl, C1-C 10 Alkoxy, 3-8 membered cycloalkyl, 3-8 membered heterocycloalkyl, and 6-8 membered aryl; and

[0376] Each R” is independently selected from hydrogen and C1-C 10 alkyl.

[0377] In some embodiments, ring B is selected from 1 to 5 R's optionally. B The 6-8 aryl groups are replaced by examples. In some embodiments, ring B is optionally replaced by 1 to 3 R groups. B Examples of substituted phenyl groups. In some embodiments, ring B is selected from those optionally replaced by 1 to 5 R groups. BExamples of substituted 5-8-membered heteroaryl groups. In some embodiments, ring B is selected from pyrrole, furanyl, furazanyl, thiophene, imidazolyl, isothiazolyl, isoxazolyl, oxazolyl, oxadiazolyl, tetrazolyl, thiazolyl, triazolyl, pyrazolyl, pyridinyl, pyrazinyl, pyridinyl, pyridonel, and pyrimidinyl, wherein each of pyrrole, furanyl, furazanyl, thiophene, imidazolyl, isothiazolyl, isoxazolyl, oxazolyl, oxadiazolyl, tetrazolyl, thiazolyl, triazolyl, pyrazolyl, pyridinyl, pyridinyl, pyridinyl, and pyrimidinyl is independently optionally replaced by 1 to 3 R groups. B Examples of [the group] are replaced. In some embodiments, ring B is selected from pyrazolyl, isothiazolyl, isoxazolyl, pyridinyl, pyrimidinyl, and thiophene, wherein each of the pyrazolyl, isothiazolyl, isoxazolyl, pyridinyl, and thiophene groups is independently and optionally selected by 1 to 3 R groups. B The instance is replaced.

[0378] In some embodiments, ring A is selected from

[0379] In some embodiments, ring A is selected from

[0380] In some embodiments, ring A is selected from

[0381] In some embodiments, ring B is selected from

[0382] In some embodiments, ring B is selected from

[0383] In some embodiments, ring B is selected from

[0384] In some embodiments, R is selected from methyl,

[0385]

[0386] In some embodiments, R is selected from methyl,

[0387] In some embodiments, the AhR antagonist according to this disclosure is a compound of formula (Ib):

[0388]

[0389] Or its pharmaceutically acceptable salt.

[0390] in:

[0391] Ring A is selected from optionally substituted heteroaryl groups and optionally substituted heterocyclic alkyl groups;

[0392] Ring B is selected from optionally substituted heteroaryl and optionally substituted heterocyclic alkyl groups; and

[0393] R is selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, optionally substituted amide, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted ester, optionally substituted heteroalkyl, optionally substituted heteroaryl, optionally substituted heterocycloalkyl, amino, cyano, halogen, hydroxyl and -C(O)H.

[0394] In some embodiments, the AhR antagonist is any of the compounds listed in Table 2.

[0395] Table 2

[0396]

[0397]

[0398]

[0399]

[0400]

[0401]

[0402]

[0403]

[0404]

[0405]

[0406]

[0407]

[0408]

[0409]

[0410]

[0411]

[0412]

[0413]

[0414]

[0415]

[0416]

[0417]

[0418]

[0419]

[0420]

[0421]

[0422]

[0423]

[0424]

[0425]

[0426]

[0427] In some embodiments, AhR antagonists are selected from the following compounds and their pharmaceutically acceptable salts:

[0428] (i)(S)-8-(5-fluoropyridin-3-yl)-3-(1-hydroxypropyl-2-yl)-6-(4-(trifluoromethoxy)phenyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0429] (ii)(S)-6-(4-chlorophenyl)-3-(1-hydroxypropyl-2-yl)-8-(6-oxo-1,6-dihydropyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0430] (iii)(S)-8-(benzo[d][1,3]dioxacyclopenten-4-yl)-6-(4-chlorophenyl)-3-(1-hydroxypropyl-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0431] (iv)(S)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-4-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0432] (v)(S)-8-(5-fluoropyridin-3-yl)-3-(1-hydroxypropyl-2-yl)-6-(4-(trifluoromethyl)phenyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0433] (vi)(S)-3-(1-hydroxypropyl-2-yl)-6,8-bis(pyridin-4-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0434] (vii)(S)-6-(4-chlorophenyl)-8-(3-fluorophenyl)-3-(1-hydroxypropyl-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0435] (viii)(S)-8-(3-fluorophenyl)-3-(1-hydroxypropyl-2-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0436] (ix)3-(2-hydroxy-2-methylpropyl)-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one;

[0437] (x)6,8-Di(pyridin-3-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0438] (xi)(S)-6-chloro-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0439] (xii)(S)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one;

[0440] (xiii)6-(4-chlorophenyl)-8-(pyridin-3-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0441] (xiv)8-(pyridin-3-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)-6-(4-(trifluoromethoxy)phenyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0442] (xv)6-(4-chlorophenyl)-8-(1-methyl-1H-pyrazol-4-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0443] (xvi)3-(2-hydroxy-2-methylpropyl)-8-(pyridin-3-yl)-6-(4-(trifluoromethyl)phenyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0444] (xvii)(S)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)-6-(p-tolyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0445] (xviii)6-(4-chlorophenyl)-3-(2-hydroxy-2-methylpropyl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0446] (xix)3-(2-hydroxy-2-methylpropyl)-6,8-bis(1-methyl-1H-pyrazol-4-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0447] (xx)(S)-3-(1-hydroxypropyl-2-yl)-6,8-bis(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0448] (xxi)6-(4-chlorophenyl)-3-(2-hydroxy-2-methylpropyl)-8-(1-methyl-1H-pyrazol-4-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0449] (xxii)(S)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)-6-(4-(trifluoromethoxy)phenyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0450] (xxiii)(S)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)-6-(4-(trifluoromethyl)phenyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0451] (xxiv)(S)-6-(4-chlorophenyl)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0452] (xxv)(S)-3-(1-hydroxypropyl-2-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(4-(trifluoromethoxy)phenyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0453] (xxvi)(S)-3-(1-hydroxypropyl-2-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-phenylpyrido[3,4-d]pyrimidin-4(3H)-one;

[0454] (xxvii)(S)-6-(4-chlorophenyl)-3-(1-hydroxypropyl-2-yl)-8-(1-methyl-1H-pyrazol-4-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0455] (xxviii)3-Methyl-8-(pyridin-3-yl)-6-(4-(trifluoromethoxy)phenyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0456] (xxix) Racemic-6-(4-chlorophenyl)-3-((trans)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0457] (xxx)(S)-6-(4-chlorophenyl)-3-(3-hydroxy-3-methylbut-2-yl)-8-(1-methyl-1H-pyrazol-4-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0458] (xxxi)(R)-6-(4-chlorophenyl)-3-(3-hydroxy-3-methylbut-2-yl)-8-(1-methyl-1H-pyrazol-4-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0459] (xxxii) Racemic-6-(4-chlorophenyl)-3-((cis)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0460] (xxxiii)(R)-6-(4-chlorophenyl)-3-(3-hydroxy-3-methylbut-2-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0461] (xxxiv)(S)-3-(3-hydroxy-3-methylbut-2-yl)-6,8-bis(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0462] (xxxv)(S)-6,8-bis(3,5-difluorophenyl)-3-(1-hydroxy-3-methylbut-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0463] (xxxvi)(S)-6-(4-chlorophenyl)-3-(3-hydroxy-3-methylbut-2-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0464] (xxxvii)(S)-8-(3,5-difluorophenyl)-3-(1-hydroxy-3-methylbut-2-yl)-6-(p-tolyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0465] (xxxviii)6-(4-chlorophenyl)-8-(3-fluorophenyl)-3-(2-hydroxy-2-methylpropyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0466] (xxxix)(R)-6-(4-chlorophenyl)-8-(3-fluorophenyl)-3-(3-hydroxy-3-methylbut-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0467] (xl)(S)-3-(1-(benzyloxy)propyl-2-yl)-8-(3-fluorophenyl)-6-(p-tolyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0468] (xli)(R)-6-(4-chlorophenyl)-8-(3-fluorophenyl)-3-(3,3,3-trifluoro-2-hydroxypropyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0469] (xlii)(S)-6-(4-chlorophenyl)-8-(3-fluorophenyl)-3-(3,3,3-trifluoro-2-hydroxypropyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0470] (xliii)(S)-3-(1-hydroxypropyl-2-yl)-6-morpholino-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0471] (xliv)(S)-3-(1-hydroxypropyl-2-yl)-8-(1H-imidazol-1-yl)-6-(4-(trifluoromethoxy)phenyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0472] (xlv)(S)-3-(1-methoxypropyl-2-yl)-8-(pyridin-3-yl)-6-(p-tolyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0473] (xlvi)(S)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0474] (xlvii)(S)-8-(3-fluorophenyl)-3-(1-hydroxypropyl-2-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0475] (xlviii)(S)-8-(3-fluorophenyl)-3-(1-hydroxypropyl-2-yl)-6-(p-tolyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0476] (xlix)(S)-8-(3-fluorophenyl)-3-(1-hydroxypropyl-2-yl)-6-(4-(trifluoromethoxy)phenyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0477] (l)(S)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)-6-(6-(trifluoromethoxy)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0478] (li)(S)-3-(2-hydroxy-2-methylpropyl)-8-(1H-pyrazol-4-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0479] (lii)(S)-5-(3-(1-hydroxypropyl-2-yl)-4-oxo-8-(pyridin-3-yl)-3,4-dihydropyridino[3,4-d]pyrimidin-6-yl)pyridinecarboxylate

[0480] (liii)(S)-3-(1-hydroxypropyl-2-yl)-6-(isothiazo-4-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0481] (liv)3-(2-hydroxy-2-methylpropyl)-8-(isothiazo-4-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0482] (lv)(S)-3-(1-hydroxypropyl-2-yl)-8-(isothiazo-4-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0483] (lvi)(S)-3-(1-hydroxypropyl-2-yl)-6,8-bis(isothiazo-4-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0484] (lvii)(S)-8-(pyridin-3-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0485] (lviii)(S)-6-(4-chlorophenyl)-3-(1-hydroxypropyl-2-yl)-8-(isothiazo-4-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0486] (lix)3-(2-hydroxy-2-methylpropyl)-6,8-bis(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0487] (lx)3-(2-hydroxy-2-methylpropyl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0488] (lxi)6-(4-chloro-2-methylphenyl)-3-(2-hydroxy-2-methylpropyl)-8-(1-methyl-1H-pyrazol-4-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0489] (lxii)(S)-3-(1-hydroxypropyl-2-yl)-8-(1H-pyrazol-4-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0490] (lxiii)(S)-3-(1-hydroxypropyl-2-yl)-8-(2-methylpyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0491] (lxiv)(S)-3-(1-hydroxypropyl-2-yl)-8-(4-methylpyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0492] (lxv)(S)-3-(1-hydroxypropyl-2-yl)-6-(4-methylthiazolyl-5-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0493] (lxvi)(S)-6-(2-Cyclopropylthiazo-5-yl)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0494] (lxvii)(S)-3-(1-hydroxypropyl-2-yl)-6-(2-isopropylthiazo-5-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0495] (lxviii)(S)-3-(1-hydroxypropyl-2-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0496] (lxix)(S)-3-(1-hydroxypropyl-2-yl)-6,8-bis(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0497] (lxx)6-(4-chlorophenyl)-3-((3S,4S)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0498] (lxxi)6-(4-chlorophenyl)-3-((3R,4R)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0499] (lxxii)3-(2-hydroxyethyl)-8-(pyridin-3-yl)-6-(4-(trifluoromethoxy)phenyl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0500] (lxxiii)(S)-6-(4-chlorophenyl)-3-(1-hydroxypropyl-2-yl)-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0501] (lxxiv)(S)-6-(6-cyclopropylpyridin-3-yl)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0502] (lxxv)(S)-3-(1-hydroxypropyl-2-yl)-6-(4-methyl-6-(trifluoromethyl)pyridin-3-yl)-8-(pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0503] (lxxvi)(S)-3-(1-hydroxypropyl-2-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0504] (lxxvii)(S)-6-(cyclohex-1-en-1-yl)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0505] (lxxviii)(S)-6,8-bis(5-fluoropyridin-3-yl)-3-(1-hydroxypropyl-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0506] (lxxix)3-((3R,4S)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0507] (lxxx)3-(2-hydroxy-2-methylpropyl)-8-(pyridin-3-yl)-6-(2-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0508] (lxxxi)6-(6-Cyclopropylpyridin-3-yl)-3-(2-hydroxy-2-methylpropyl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0509] (lxxxii)6-(6-Cyclopropylpyridin-3-yl)-3-(2-hydroxy-2-methylpropyl)-8-(1-methyl-1H-pyrazol-4-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0510] (lxxxiii)(S)-6-(6-cyclopropylpyridin-3-yl)-3-(1-hydroxypropyl-2-yl)-8-(1-methyl-1H-pyrazol-4-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0511] (lxxxiv)(S)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)-6-(thiazo-5-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0512] (lxxxv)(S)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)-6-(2-(trifluoromethyl)thiazo-5-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0513] (lxxxvi)(S)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)-6-(2-(trifluoromethyl)pyrimidin-5-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0514] (lxxxvii)3-(2-hydroxy-2-methylpropyl)-8-(pyridin-3-yl)-6-(2-(trifluoromethyl)thiazo-5-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0515] (lxxxviii)3-((3S,4S)-4-hydroxytetrahydrofuran-3-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0516] (lxxxix)(S)-3-(1-hydroxypropyl-2-yl)-6-(2-methylthiazolyl-5-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0517] (xc)(S)-6-(4-chlorophenyl)-8-(3-fluorophenyl)-3-(1-hydroxy-3-methylbut-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0518] (xci)(S)-3-(1-hydroxypropyl-2-yl)-6-(piperidin-1-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0519] (xcii)3-(2-hydroxy-2-methylpropyl)-8-(isothiazo-4-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0520] (xciii)(S)-3-(1-hydroxypropyl-2-yl)-8-(isothiazo-4-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0521] (xciv)3-(2-hydroxy-2-methylpropyl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0522] (xcv)(S)-8-(pyridin-3-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0523] (xcvi)(3-(2-hydroxy-2-methylpropyl)-8-(pyridin-3-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0524] (xcvii)3-(1,1-tetrahydrothiophen-3-yl)-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0525] (xcviii)(R)-3-(1,1-tetrahydrothiophene-3-yl)-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0526] (xcix)(R)-3-(2-hydroxypropyl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0527] (c)(S)-3-(2-hydroxypropyl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0528] (ci)(R)-8-(1-methyl-1H-pyrazol-4-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0529] (cii)(S)-8-(1-methyl-1H-pyrazol-4-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0530] (ciii)(R)-3-(2-hydroxypropyl)-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0531] (civ)(R)-8-(1-methyl-1H-pyrazol-4-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0532] (cv)3-((3S,4R)-4-hydroxytetrahydrofuran-3-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0533] (cvi)3-((3R,4S)-4-hydroxytetrahydrofuran-3-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0534] (cvii)6-(4-chlorophenyl)-3-((3S,4R)-4-hydroxytetrahydrofuran-3-yl)-8-(1-methyl-1H-pyrazol-4-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0535] (cviii)3-((3S,4R)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0536] (cix)(S)-6-(6-cyclopropylpyridin-3-yl)-8-(pyridin-3-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0537] (cx)(R)-6-(6-cyclopropylpyridin-3-yl)-8-(pyridin-3-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0538] (cxi)(R)-8-(pyridin-3-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0539] (cxii)(R)-8-(pyridin-3-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)-6-(4-(trifluoromethoxy)phenyl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0540] (cxiii)(S)-8-(pyridin-3-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)-6-(4-(trifluoromethoxy)phenyl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0541] (cxiv)6-(4-chlorophenyl)-3-((3S,4R)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0542] (cxv)3-((3S,4S)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0543] Methyl (cxvi)(S)-2-(4-oxo-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-3(4H)-yl)propionate

[0544] (cxvii)6-(4-chlorophenyl)-3-(4-hydroxy-1-methylpyrrolidin-3-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0545] (cxviii)6-(4-chlorophenyl)-3-((3R,4R)-4-hydroxypyrrolidin-3-yl)-8-(pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0546] (cxix)(R)-6-(6-cyclopropylpyridin-3-yl)-8-(1-methyl-1H-pyrazol-4-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0547] (cxx)(S)-6-(6-cyclopropylpyridin-3-yl)-8-(1-methyl-1H-pyrazol-4-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0548] (cxxi)3-((3S,4R)-4-hydroxytetrahydrofuran-3-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0549] (cxxii)3-((3R,4S)-4-hydroxytetrahydrofuran-3-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0550] (cxxiii)3-((3S,4R)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0551] (cxxiv)3-((3R,4S)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0552] (cxxv)(R)-3-(2-hydroxypropyl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0553] (cxxvi)(S)-3-(2-hydroxypropyl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0554] (cxxvii)(S)-3-(2-hydroxypropyl)-8-(pyridin-3-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0555] (cxxviii)(R)-3-(2-hydroxypropyl)-8-(pyridin-3-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0556] (cxxix)3-((3S,4S)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0557] (cxxx)(S)-3-(2-hydroxypropyl)-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0558] (cxxxi)(S)-8-(1-methyl-1H-pyrazol-4-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0559] (cxxxii)(S)-3-(1-hydroxypropyl-2-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0560] (cxxxiii)3-((3S,4S)-4-hydroxytetrahydrofuran-3-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0561] (cxxxiv)6-(4-chlorophenyl)-3-((3R,4S)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0562] (cxxxv)6-(4-chlorophenyl)-3-((3R,4S)-4-hydroxytetrahydrofuran-3-yl)-8-(1-methyl-1H-pyrazol-4-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0563] (cxxxvi)(R)-8-(pyridin-3-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0564] (cxxxvii)(S)-2-(4-oxo-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-3(4H)-yl)propionic acid

[0565] (cxxxviii)(S)-N-methyl-2-(4-oxo-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-3(4H)-yl)propionamide

[0566] (cxxxix)(S)-N,N-dimethyl-2-(4-oxo-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-3(4H)-yl)acrylamide

[0567] (cxl)3-(2-hydroxy-2-methylpropyl)-8-(1H-pyrazol-4-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0568] (cxli)3-(2-hydroxy-2-methylpropyl)-8-(1H-imidazol-1-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0569] (cxlii)3-(2-hydroxy-2-methylpropyl)-8-(1H-imidazol-1-yl)-6-(6-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0570] (cxliii)(S)-3-(1-hydroxypropyl-2-yl)-8-(1H-imidazol-1-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0571] (cxliv)(S)-3-(1-hydroxypropyl-2-yl)-8-(1H-imidazol-1-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0572] (cxlv)(S)-3-(1-hydroxypropyl-2-yl)-8-(1H-1,2,4-triazol-1-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0573] (cxlvi)(S)-3-(1-hydroxypropyl-2-yl)-8-(1H-pyrazol-1-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0574] (cxlvii)(S)-3-(1-hydroxypropyl-2-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(2-(trifluoromethyl)thiazo-5-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0575] (cxlviii)(S)-8-(diethylamino)-3-(1-hydroxypropyl-2-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0576] (cxlix)(S)-3-(1-hydroxypropyl-2-yl)-8-(piperidin-1-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0577] (cl)(S)-3-(1-hydroxypropyl-2-yl)-8-(pyrrolid-1-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0578] (cli)(S)-6-(4-chlorophenyl)-3-(1-hydroxypropyl-2-yl)-8-(piperidin-1-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0579] (clii)(S)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-2-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0580] (cliii)(S)-6-cyclohexyl-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0581] (cliv)(S)-3-(1-hydroxypropyl-2-yl)-6-(pyridin-2-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0582] (clv)(S)-3-(1-hydroxypropyl-2-yl)-6-(2-methylthiazo-4-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0583] (clvi)(S)-3-(1-hydroxypropyl-2-yl)-6-(1-methyl-1H-1,2,3-triazol-5-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0584] (clvii)(R)-6-(4-chlorophenyl)-8-(pyridin-3-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0585] (clviii)(S)-6-(4-chlorophenyl)-8-(pyridin-3-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0586] (clix)(S)-3-(1-hydroxypropyl-2-yl)-8-(1-methyl-1,2,5,6-tetrahydropyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0587] (clx)6-(4-chlorophenyl)-3-((3S,4S)-4-hydroxytetrahydrofuran-3-yl)-8-(1-methyl-1H-pyrazol-4-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0588] (clxi)(S)-3-(1-hydroxypropyl-2-yl)-6-(2-methylpyrimidin-5-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0589] (clxii)3-(2-hydroxy-2-methylpropyl)-8-(1-(trifluoromethyl)-1H-pyrazol-4-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0590] (clxiii)(S)-3-(1-hydroxypropyl-2-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(2-(trifluoromethyl)pyrimidin-5-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0591] (clxiv)3-(2-hydroxy-2-methylpropyl)-8-(pyridin-3-yl)-6-(2-(trifluoromethyl)pyrimidin-5-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0592] (clxv)(S)-5-(3-(1-hydroxypropyl-2-yl)-4-oxo-8-(pyridin-3-yl)-3,4-dihydropyridino[3,4-d]pyrimidin-6-yl)pyridinecarboxylic acid

[0593] (clxvi)(S)-3-(1-hydroxypropyl-2-yl)-6-(6-methylpyridin-3-yl)-8-(pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0594] (clxvii)3-(2-hydroxy-2-methylpropyl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(2-(trifluoromethyl)pyrimidin-5-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0595] (clxviii)3,8-Di(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0596] (clxix)8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0597] (clxx)3-((3R,4R)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0598] (clxxi)3-((3R,4R)-4-hydroxytetrahydrofuran-3-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0599] (clxxii)6-(4-chlorophenyl)-3-((3R,4R)-4-hydroxytetrahydrofuran-3-yl)-8-(1-methyl-1H-pyrazol-4-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0600] (clxxiii)3-Cyclopentyl-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0601] (clxxiv)3-Phenyl-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0602] (clxxv)3-((3R,4R)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0603] (clxxvi)3-((3R,4R)-4-hydroxytetrahydrofuran-3-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0604] (clxxvii)(S)-3-(1-hydroxypropyl-2-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0605] (clxxviii)(S)-N-((3R,4S)-4-hydroxytetrahydrofuran-3-yl)-2-(4-oxo-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-3(4H)-yl)propionamide

[0606] (clxxix)3-(2-hydroxy-2-methylpropyl)-8-(1H-pyrazol-4-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0607] (clxxx)(S)-6-(4-chlorophenyl)-3-(1-hydroxypropyl-2-yl)-8-(1H-pyrazol-1-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0608] (clxxxi)(S)-3-(1-hydroxypropyl-2-yl)-8-(1H-pyrazol-1-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0609] (clxxxii)3-((3S,4R)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)-6-(2-(trifluoromethyl)thiazo-5-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0610] (clxxxiii)3-(2-hydroxy-2-methylpropyl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(2-(trifluoromethyl)thiazo-5-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0611] (clxxxiv)3-((3S,4R)-4-hydroxytetrahydrofuran-3-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(2-(trifluoromethyl)thiazo-5-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0612] (clxxxv)3-((3R,4S)-4-hydroxytetrahydrofuran-3-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(2-(trifluoromethyl)thiazo-5-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0613] (clxxxvi)3-((3R,4S)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)-6-(2-(trifluoromethyl)thiazo-5-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0614] (clxxxvii)(S)-3-(1-hydroxypropyl-2-yl)-8-morpholino-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0615] (clxxxviii)3-(2-hydroxy-2-methylpropyl)-8-(piperidin-1-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0616] (clxxxix)(S)-3-(1-hydroxypropyl-2-yl)-6-(5-methylpyridin-2-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0617] (cxc)(S)-3-(1-hydroxypropyl-2-yl)-6-(5-methylpyrimidin-2-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0618] (cxci)(S)-8-(cyclohex-1-en-1-yl)-3-(1-hydroxypropyl-2-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0619] (cxcii)(S)-8-cyclohexyl-3-(1-hydroxypropyl-2-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0620] (cxciii)(S)-3-(1-hydroxypropyl-2-yl)-N,N-dimethyl-4-oxo-8-(pyridin-3-yl)-3,4-dihydropyrido[3,4-d]pyrimidin-6-carboxamide

[0621] (cxciv)(S)-3-(1-hydroxypropyl-2-yl)-8-(1H-pyrazol-4-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0622] (cxcv)(S)-3-(1-hydroxypropyl-2-yl)-6-(2-methoxyethyl)-8-(pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0623] (cxcvi)(S)-3-(1-hydroxypropyl-2-yl)-8-(2-methoxyethyl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one.

[0624] In some embodiments, AhR antagonists are selected from the following compounds and their pharmaceutically acceptable salts:

[0625] (i)(S)-8-(5-fluoropyridin-3-yl)-3-(1-hydroxypropyl-2-yl)-6-(4-(trifluoromethoxy)phenyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0626] (ii)(S)-6-(4-chlorophenyl)-3-(1-hydroxypropyl-2-yl)-8-(6-oxo-1,6-dihydropyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0627] (iii)(S)-8-(benzo[d][1,3]dioxacyclopenten-4-yl)-6-(4-chlorophenyl)-3-(1-hydroxypropyl-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0628] (iv)(S)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-4-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0629] (v)(S)-8-(5-fluoropyridin-3-yl)-3-(1-hydroxypropyl-2-yl)-6-(4-(trifluoromethyl)phenyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0630] (vi)(S)-3-(1-hydroxypropyl-2-yl)-6,8-bis(pyridin-4-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0631] (vii)(S)-6-(4-chlorophenyl)-8-(3-fluorophenyl)-3-(1-hydroxypropyl-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0632] (viii)(S)-8-(3-fluorophenyl)-3-(1-hydroxypropyl-2-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0633] (ix)3-(2-hydroxy-2-methylpropyl)-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one;

[0634] (x)6,8-Di(pyridin-3-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0635] (xi)(S)-6-chloro-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0636] (xii)(S)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one;

[0637] (xiii)6-(4-chlorophenyl)-8-(pyridin-3-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0638] (xiv)8-(pyridin-3-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)-6-(4-(trifluoromethoxy)phenyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0639] (xv)6-(4-chlorophenyl)-8-(1-methyl-1H-pyrazol-4-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0640] (xvi)3-(2-hydroxy-2-methylpropyl)-8-(pyridin-3-yl)-6-(4-(trifluoromethyl)phenyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0641] (xvii)(S)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)-6-(p-tolyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0642] (xviii)6-(4-chlorophenyl)-3-(2-hydroxy-2-methylpropyl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0643] (xix)3-(2-hydroxy-2-methylpropyl)-6,8-bis(1-methyl-1H-pyrazol-4-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0644] (xx)(S)-3-(1-hydroxypropyl-2-yl)-6,8-bis(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0645] (xxi)6-(4-chlorophenyl)-3-(2-hydroxy-2-methylpropyl)-8-(1-methyl-1H-pyrazol-4-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0646] (xxii)(S)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)-6-(4-(trifluoromethoxy)phenyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0647] (xxiii)(S)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)-6-(4-(trifluoromethyl)phenyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0648] (xxiv)(S)-6-(4-chlorophenyl)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0649] (xxv)(S)-3-(1-hydroxypropyl-2-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(4-(trifluoromethoxy)phenyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0650] (xxvi)(S)-3-(1-hydroxypropyl-2-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-phenylpyrido[3,4-d]pyrimidin-4(3H)-one;

[0651] (xxvii)(S)-6-(4-chlorophenyl)-3-(1-hydroxypropyl-2-yl)-8-(1-methyl-1H-pyrazol-4-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0652] (xxviii)3-Methyl-8-(pyridin-3-yl)-6-(4-(trifluoromethoxy)phenyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0653] (xxix) Racemic-6-(4-chlorophenyl)-3-((trans)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0654] (xxx)(S)-6-(4-chlorophenyl)-3-(3-hydroxy-3-methylbut-2-yl)-8-(1-methyl-1H-pyrazol-4-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0655] (xxxi)(R)-6-(4-chlorophenyl)-3-(3-hydroxy-3-methylbut-2-yl)-8-(1-methyl-1H-pyrazol-4-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0656] (xxxii) Racemic-6-(4-chlorophenyl)-3-((cis)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0657] (xxxiii)(R)-6-(4-chlorophenyl)-3-(3-hydroxy-3-methylbut-2-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0658] (xxxiv)(S)-3-(3-hydroxy-3-methylbut-2-yl)-6,8-bis(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0659] (xxxv)(S)-6,8-bis(3,5-difluorophenyl)-3-(1-hydroxy-3-methylbut-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0660] (xxxvi)(S)-6-(4-chlorophenyl)-3-(3-hydroxy-3-methylbut-2-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0661] (xxxvii)(S)-8-(3,5-difluorophenyl)-3-(1-hydroxy-3-methylbut-2-yl)-6-(p-tolyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0662] (xxxviii)6-(4-chlorophenyl)-8-(3-fluorophenyl)-3-(2-hydroxy-2-methylpropyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0663] (xxxix)(R)-6-(4-chlorophenyl)-8-(3-fluorophenyl)-3-(3-hydroxy-3-methylbut-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0664] (xl)(S)-3-(1-(benzyloxy)propyl-2-yl)-8-(3-fluorophenyl)-6-(p-tolyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0665] (xli)(R)-6-(4-chlorophenyl)-8-(3-fluorophenyl)-3-(3,3,3-trifluoro-2-hydroxypropyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0666] (xlii)(S)-6-(4-chlorophenyl)-8-(3-fluorophenyl)-3-(3,3,3-trifluoro-2-hydroxypropyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0667] (xliii)(S)-3-(1-hydroxypropyl-2-yl)-6-morpholino-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0668] (xliv)(S)-3-(1-hydroxypropyl-2-yl)-8-(1H-imidazol-1-yl)-6-(4-(trifluoromethoxy)phenyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0669] (xlv)(S)-3-(1-methoxypropyl-2-yl)-8-(pyridin-3-yl)-6-(p-tolyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0670] (xlvi)(S)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0671] (xlvii)(S)-8-(3-fluorophenyl)-3-(1-hydroxypropyl-2-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0672] (xlviii)(S)-8-(3-fluorophenyl)-3-(1-hydroxypropyl-2-yl)-6-(p-tolyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0673] (xlix)(S)-8-(3-fluorophenyl)-3-(1-hydroxypropyl-2-yl)-6-(4-(trifluoromethoxy)phenyl)pyrido[3,4-d]pyrimidin-4(3H)-one;

[0674] (l)(S)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)-6-(6-(trifluoromethoxy)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0675] (li)(S)-3-(2-hydroxy-2-methylpropyl)-8-(1H-pyrazol-4-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0676] (lii)(S)-5-(3-(1-hydroxypropyl-2-yl)-4-oxo-8-(pyridin-3-yl)-3,4-dihydropyridino[3,4-d]pyrimidin-6-yl)pyridinecarboxylate

[0677] (liii)(S)-3-(1-hydroxypropyl-2-yl)-6-(isothiazo-4-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0678] (liv)3-(2-hydroxy-2-methylpropyl)-8-(isothiazo-4-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0679] (lv)(S)-3-(1-hydroxypropyl-2-yl)-8-(isothiazo-4-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0680] (lvi)(S)-3-(1-hydroxypropyl-2-yl)-6,8-bis(isothiazo-4-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0681] (lvii)(S)-8-(pyridin-3-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0682] (lviii)(S)-6-(4-chlorophenyl)-3-(1-hydroxypropyl-2-yl)-8-(isothiazo-4-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0683] (lix)3-(2-hydroxy-2-methylpropyl)-6,8-bis(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0684] (lx)3-(2-hydroxy-2-methylpropyl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0685] (lxi)6-(4-chloro-2-methylphenyl)-3-(2-hydroxy-2-methylpropyl)-8-(1-methyl-1H-pyrazol-4-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0686] (lxii)(S)-3-(1-hydroxypropyl-2-yl)-8-(1H-pyrazol-4-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0687] (lxiii)(S)-3-(1-hydroxypropyl-2-yl)-8-(2-methylpyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0688] (lxiv)(S)-3-(1-hydroxypropyl-2-yl)-8-(4-methylpyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0689] (lxv)(S)-3-(1-hydroxypropyl-2-yl)-6-(4-methylthiazolyl-5-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0690] (lxvi)(S)-6-(2-Cyclopropylthiazo-5-yl)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0691] (lxvii)(S)-3-(1-hydroxypropyl-2-yl)-6-(2-isopropylthiazo-5-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0692] (lxviii)(S)-3-(1-hydroxypropyl-2-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0693] (lxix)(S)-3-(1-hydroxypropyl-2-yl)-6,8-bis(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0694] (lxx)6-(4-chlorophenyl)-3-((3S,4S)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0695] (lxxi)6-(4-chlorophenyl)-3-((3R,4R)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0696] (lxxii)3-(2-hydroxyethyl)-8-(pyridin-3-yl)-6-(4-(trifluoromethoxy)phenyl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0697] (lxxiii)(S)-6-(4-chlorophenyl)-3-(1-hydroxypropyl-2-yl)-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0698] (lxxiv)(S)-6-(6-cyclopropylpyridin-3-yl)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0699] (lxxv)(S)-3-(1-hydroxypropyl-2-yl)-6-(4-methyl-6-(trifluoromethyl)pyridin-3-yl)-8-(pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0700] (lxxvi)(S)-3-(1-hydroxypropyl-2-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0701] (lxxvii)(S)-6-(cyclohex-1-en-1-yl)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0702] (lxxviii)(S)-6,8-bis(5-fluoropyridin-3-yl)-3-(1-hydroxypropyl-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0703] (lxxix)3-((3R,4S)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0704] (lxxx)3-(2-hydroxy-2-methylpropyl)-8-(pyridin-3-yl)-6-(2-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0705] (lxxxi)6-(6-Cyclopropylpyridin-3-yl)-3-(2-hydroxy-2-methylpropyl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0706] (lxxxii)6-(6-Cyclopropylpyridin-3-yl)-3-(2-hydroxy-2-methylpropyl)-8-(1-methyl-1H-pyrazol-4-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0707] (lxxxiii)(S)-6-(6-cyclopropylpyridin-3-yl)-3-(1-hydroxypropyl-2-yl)-8-(1-methyl-1H-pyrazol-4-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0708] (lxxxiv)(S)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)-6-(thiazo-5-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0709] (lxxxv)(S)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)-6-(2-(trifluoromethyl)thiazo-5-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0710] (lxxxvi)(S)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)-6-(2-(trifluoromethyl)pyrimidin-5-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0711] (lxxxvii)3-(2-hydroxy-2-methylpropyl)-8-(pyridin-3-yl)-6-(2-(trifluoromethyl)thiazo-5-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0712] (lxxxviii)3-((3S,4S)-4-hydroxytetrahydrofuran-3-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0713] (lxxxix)(S)-3-(1-hydroxypropyl-2-yl)-6-(2-methylthiazolyl-5-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0714] (xc)(S)-6-(4-chlorophenyl)-8-(3-fluorophenyl)-3-(1-hydroxy-3-methylbut-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0715] (xci)(S)-3-(1-hydroxypropyl-2-yl)-6-(piperidin-1-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0716] (xcii)3-(2-hydroxy-2-methylpropyl)-8-(isothiazo-4-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0717] (xciii)(S)-3-(1-hydroxypropyl-2-yl)-8-(isothiazo-4-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0718] (xciv)3-(2-hydroxy-2-methylpropyl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0719] (xcv)(S)-8-(pyridin-3-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0720] (xcvi)(3-(2-hydroxy-2-methylpropyl)-8-(pyridin-3-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0721] (xcvii)3-(1,1-tetrahydrothiophen-3-yl)-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0722] (xcviii)(R)-3-(1,1-tetrahydrothiophene-3-yl)-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0723] (xcix)(R)-3-(2-hydroxypropyl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0724] (c)(S)-3-(2-hydroxypropyl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0725] (ci)(R)-8-(1-methyl-1H-pyrazol-4-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0726] (cii)(S)-8-(1-methyl-1H-pyrazol-4-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0727] (ciii)(R)-3-(2-hydroxypropyl)-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0728] (civ)(R)-8-(1-methyl-1H-pyrazol-4-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0729] (cv)3-((3S,4R)-4-hydroxytetrahydrofuran-3-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0730] (cvi)3-((3R,4S)-4-hydroxytetrahydrofuran-3-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0731] (cvii)6-(4-chlorophenyl)-3-((3S,4R)-4-hydroxytetrahydrofuran-3-yl)-8-(1-methyl-1H-pyrazol-4-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0732] (cviii)3-((3S,4R)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0733] (cix)(S)-6-(6-cyclopropylpyridin-3-yl)-8-(pyridin-3-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0734] (cx)(R)-6-(6-cyclopropylpyridin-3-yl)-8-(pyridin-3-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0735] (cxi)(R)-8-(pyridin-3-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0736] (cxii)(R)-8-(pyridin-3-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)-6-(4-(trifluoromethoxy)phenyl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0737] (cxiii)(S)-8-(pyridin-3-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)-6-(4-(trifluoromethoxy)phenyl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0738] (cxiv)6-(4-chlorophenyl)-3-((3S,4R)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0739] (cxv)3-((3S,4S)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0740] Methyl (cxvi)(S)-2-(4-oxo-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-3(4H)-yl)propionate

[0741] (cxvii)6-(4-chlorophenyl)-3-(4-hydroxy-1-methylpyrrolidin-3-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0742] (cxviii)6-(4-chlorophenyl)-3-((3R,4R)-4-hydroxypyrrolidin-3-yl)-8-(pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0743] (cxix)(R)-6-(6-cyclopropylpyridin-3-yl)-8-(1-methyl-1H-pyrazol-4-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0744] (cxx)(S)-6-(6-cyclopropylpyridin-3-yl)-8-(1-methyl-1H-pyrazol-4-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0745] (cxxi)3-((3S,4R)-4-hydroxytetrahydrofuran-3-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0746] (cxxii)3-((3R,4S)-4-hydroxytetrahydrofuran-3-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0747] (cxxiii)3-((3S,4R)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0748] (cxxiv)3-((3R,4S)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0749] (cxxv)(R)-3-(2-hydroxypropyl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0750] (cxxvi)(S)-3-(2-hydroxypropyl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0751] (cxxvii)(S)-3-(2-hydroxypropyl)-8-(pyridin-3-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0752] (cxxviii)(R)-3-(2-hydroxypropyl)-8-(pyridin-3-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0753] (cxxix)3-((3S,4S)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0754] (cxxx)(S)-3-(2-hydroxypropyl)-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0755] (cxxxi)(S)-8-(1-methyl-1H-pyrazol-4-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0756] (cxxxii)(S)-3-(1-hydroxypropyl-2-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0757] (cxxxiii)3-((3S,4S)-4-hydroxytetrahydrofuran-3-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0758] (cxxxiv)6-(4-chlorophenyl)-3-((3R,4S)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0759] (cxxxv)6-(4-chlorophenyl)-3-((3R,4S)-4-hydroxytetrahydrofuran-3-yl)-8-(1-methyl-1H-pyrazol-4-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0760] (cxxxvi)(R)-8-(pyridin-3-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0761] (cxxxvii)(S)-2-(4-oxo-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-3(4H)-yl)propionic acid

[0762] (cxxxviii)(S)-N-methyl-2-(4-oxo-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-3(4H)-yl)propionamide

[0763] (cxxxix)(S)-N,N-dimethyl-2-(4-oxo-8-(pyridin-3-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-3(4H)-yl)acrylamide

[0764] (cxl)3-(2-hydroxy-2-methylpropyl)-8-(1H-pyrazol-4-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0765] (cxli)3-(2-hydroxy-2-methylpropyl)-8-(1H-imidazol-1-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0766] (cxlii)3-(2-hydroxy-2-methylpropyl)-8-(1H-imidazol-1-yl)-6-(6-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0767] (cxliii)(S)-3-(1-hydroxypropyl-2-yl)-8-(1H-imidazol-1-yl)-6-(5-(trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0768] (cxliv)(S)-3-(1-hydroxypropyl-2-yl)-8-(1H-imidazol-1-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0769] (cxlv)(S)-3-(1-hydroxypropyl-2-yl)-8-(1H-1,2,4-triazol-1-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0770] (cxlvi)(S)-3-(1-hydroxypropyl-2-yl)-8-(1H-pyrazol-1-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0771] (cxlvii)(S)-3-(1-hydroxypropyl-2-yl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(2-(trifluoromethyl)thiazo-5-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0772] (cxlviii)(S)-8-(diethylamino)-3-(1-hydroxypropyl-2-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0773] (cxlix)(S)-3-(1-hydroxypropyl-2-yl)-8-(piperidin-1-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0774] (cl)(S)-3-(1-hydroxypropyl-2-yl)-8-(pyrrolid-1-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one

[0775] (cli)(S)-6-(4-chlorophenyl)-3-(1-hydroxypropyl-2-yl)-8-(piperidin-1-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0776] (clii)(S)-3-(1-hydroxypropyl-2-yl)-8-(pyridin-2-yl)-6-(6-(trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0777] (cliii)(S)-6-cyclohexyl-3-(1-hydroxypropyl-2-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0778] (cliv)(S)-3-(1-hydroxypropyl-2-yl)-6-(pyridin-2-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0779] (clv)(S)-3-(1-hydroxypropyl-2-yl)-6-(2-methylthiazo-4-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0780] (clvi)(S)-3-(1-hydroxypropyl-2-yl)-6-(1-methyl-1H-1,2,3-triazol-5-yl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0781] (clvii)(R)-6-(4-chlorophenyl)-8-(pyridin-3-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)pyrido[3,4-d]pyrimidin-4(3H)-one

[0782] (clviii)(S)-6-(4-chlorophenyl)-8-(pyridin-3-yl)-3-(3,3,3-trifluoro-2-hydroxypropyl)pyrido[3,4-d]pyrimidin-4(3H)-one.

[0783] Example

[0784] The synthesis of AhR antagonists and the biological data of these compounds described in Table 2 are described in PCT International Publication No. WO2021 / 102288 A1, the contents of which are incorporated herein by reference in their entirety.

[0785] Example 1: AhR ligands for predicting ICI responsiveness

[0786] An experimental cohort of patients with advanced (III-IV) non-small cell lung cancer (donors) was recruited to participate in a study evaluating the role of the microbiome in the response to immunomodulatory therapy (ICI). Therapies evaluated included any single or combined use of PD-1, PD-L1, and CLTA-4-targeted therapies. Samples collected to characterize the effect of the microbiome on patient response consisted of paired fecal and plasma samples, collected at two time points: (i) before therapy initiation and (ii) approximately six weeks after therapy initiation. Patients were monitored for up to three years post-recruitment to establish response to administered immunomodulatory therapy based on adopted RECIST 1.1 criteria. A total of 73 patients met the requirements for this study and provided samples.

[0787] Fecal (intestinal lumen) and plasma samples collected from all donors at all time points were processed in parallel to characterize signals from metabolites shared across the microbiome and host. Samples were stored according to best practices in microbiome science.

[0788] Metabolite concentrations were measured using targeted GC / MS, LC / MS, and LC / MS / MS platforms, including ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS / MS), as described by Zierer et al. (Nat Genet. [Nature Genetics], 2018, 50(6):790-795), Metabolon, Inc., North Carolina, USA. The following instruments / conditions were used: a Waters ACQUITY ultra-high performance liquid chromatograph (UPLC) and a Thermo Scientific Q-Exactive high-resolution / precision mass spectrometer interoperable with a heated electrospray ionization (HESI-II) source and an Orbitrap mass analyzer operating at 35,000 mass resolution. Sample extracts were dried and then reconstituted in solvents compatible with each of the four aliquots determined below. Each reconstitution solvent contained a series of standards at fixed concentrations to ensure consistency between injection and chromatography.

[0789] The first aliquot was analyzed under acidic cation conditions, with chromatographic optimization for more hydrophilic compounds. The extract was eluted from a C18 column (Waters UPLC BEH C18-2.1 x 100 mm, 1.7 μm) using a gradient elution with water and methanol containing 0.05% perfluoropentanoic acid (PFPA) and 0.1% formic acid (FA). The second aliquot was also analyzed under acidic cation conditions, but it was chromatographically optimized for more hydrophobic compounds. In this determination, the extract was eluted from the same C18 column using a gradient elution with methanol, acetonitrile, water, 0.05% PFPA, and 0.01% FA, and operated at a higher overall organic content. The third aliquot was analyzed using a separate dedicated C18 column with optimized alkaline negative ion conditions. The alkaline extract was eluted from the column using a gradient of methanol and water (containing 6.5 mM ammonium bicarbonate at pH 8). After elution from a HILIC column (Waters UPLC BEH Amide 2.1 x 150 mm, 1.7 μm), the fourth aliquot was analyzed using negative ionization with a gradient of water and acetonitrile (containing 10 mM ammonium formate, pH 10.8). MS analysis used dynamic exclusion, alternating between MS and data-dependent MSn scans. Scan ranges varied slightly between determinations but covered 70–1000 m / z. Raw data files were archived and extracted by Metabolon Inc.

[0790] From the generated small molecule distribution data, the target group of 29 compounds known to function as AhR ligands was investigated (Table 1). A binary representation of the response to ICI therapy was constructed for each patient using metadata describing patient outcomes. AhR ligands in both fecal and plasma samples were further analyzed using a statistical association test (MannWhitney U with false discovery rate correction) at the baseline time point prior to the actual initiation of therapy to assess any potential for such ligands to have a predictive or predetermined role in the success of ICI therapy. Figure 1A As shown, four ligands, namely kynurenine, tryptophan, phenylpyruvic acid, and indolelactate, were found to be highly differentiated at baseline, with FDR-corrected p-values ​​less than 0.05 when compared between patients who eventually became responders or non-responders.

[0791] Of these four AhR ligands, all were statistically associated with future response in this clinical study. All of these ligands were found to be elevated on average in non-responders compared to responders, suggesting that the observed absence of these AhR ligands may indicate a higher probability of response to ICI therapy.

[0792] like Figure 1BAs shown, using the same method, the corresponding signals of these ligands in paired plasma samples from these donors did not show statistical differences, indicating that the elevation of these AhR ligands in patients who eventually became non-responders could be observed simply by directly measuring fecal samples rather than plasma samples.

[0793] Although only a small clinical group (19 out of 73) of patients who received ICI alone without other concomitant therapies (such as chemotherapy) also demonstrated this significant shift in the levels of these four AhR ligands, indicating potential future non-responsiveness to ICI therapy ( Figure 2 The data further demonstrate the correlation between the microbial metabolism of these AhR ligands and the prediction of successful treatment response to ICI therapy.

[0794] Signals from 29 AhR ligands across the entire graph were compared with four statistically correlated ligands, allowing for visual observation of these signals in future responders or non-responders, as shown by principal component analysis used for dimensionality reduction. Figure 3 As shown in the left and middle figures, the grouping of responders qualitatively increases as the view moves from the full graph to a visualization of signals from only the four relevant ligands. When comparing this view of relevant ligands only from the ICI donor, the distinction between responders and non-responders becomes even more significant (right figure), confirming the potential predictive or deterministic nature of these four AhR ligands in successful ICI therapy.

[0795] Example 2: DRE-luciferase reporter gene assay

[0796] AhR binds to dioxin-responsive elements (DREs) upstream of the activated gene. One measure of AhR activity is the activation of reporter genes downstream of one or more DRE elements, such as luciferase. Luciferase activity reflects AhR activation and inhibition in cells expressing the reporter gene. 20,000 human HepG2 liver cancer-AhR-Lucia reporter cells or human HT29 colon adenocarcinoma-AhR reporter cells or other stably transfected DRE-luciferase reporter gene cell lines were plated in Eagle's minimum essential medium, 10% heat-inactivated FBS, 1X non-essential amino acid Pen-Strep (10,000 U / mL), and Normocin (100 μg / mL) in 96-well, 384-well, or other plates and incubated overnight at 37°C in a CO2 incubator, with and without AhR antagonists at logarithmic dilutions starting at 100 μM.

[0797] One hour after seeding the cell plates, AhR activating ligands such as TCDD, kynurenine, ITE (methyl 2-(lH-indol-3-ylcarbonyl)-4-thiazolylcarboxylate), VAF347, BNF (β-naphthylflavonoid), FICZ (6-formylindol(3,2-b)carbazole), or other AhR ligands were applied at their specific EC50 values. 50 The concentration was added to cells with or without AhR antagonists.

[0798] Cells were incubated for 24 or 48 hours or at another time point, and the supernatant was analyzed to determine luciferase activity as a reading of AhR activation or inhibition. The commercial QUANTI-Luc kit from InvivoGen was used according to the manufacturer's instructions. TM The assay kit measures luciferase.

[0799] The level of luciferase was at its maximum when only an agonist ligand was added, while it was at its minimum when no antagonist was added. IC 50 The value was determined to be the concentration that inhibits half of the luciferase activity. The IC50 of the novel AhR antagonist for luciferase disclosed herein... 50 The level report is in Table 3. "A" indicates IC. 50 Value less than 100nM, "B" indicates IC 50 Between 100 and 500 nm, "C" indicates the IC. 50 Above 500nM, and "D" indicates IC 50 The value cannot be generated from the data.

[0800] Table 3

[0801]

[0802]

[0803]

[0804]

[0805]

[0806]

[0807]

[0808] Example 3: Combination Therapy

[0809] The antitumor efficacy of compound 30 was investigated in a CT26 mouse model unresponsive to PD-L1 therapy alone. CT26 cells were subcutaneously implanted into Balb / c mice, which were then randomized and treated with an isotope-controlled antibody, an anti-PD-L1 antibody, or a combination of compound 30 (Table 2) and an anti-PD-L1 antibody. Compound 30 was administered orally at 10 mg / kg once daily for 14 days, and the anti-PD-L1 antibody was administered intraperitoneally at 10 mg / kg every 3 days. Tumor growth curves and tumor weights for all treatment groups are shown in the figure. Figure 4A and 4B As shown in the figure. Although the anti-PD-L1 antibody alone did not significantly affect tumor growth or tumor weight, co-administration of compound 30 with the anti-PD-L1 antibody resulted in a reduction in tumor volume, p = 0.039. Figure 4A (Mann-Whitney, nonparametric test). Additionally, co-administration of compound 30 with the anti-PD-L1 antibody resulted in a reduction in tumor weight, p = 0.067. Figure 4B (Mann-Whitney, a nonparametric test).

[0810] Although this disclosure has been specifically shown and described with reference to particular embodiments, those skilled in the art will understand that various changes in form and detail may be made therein without departing from the spirit and scope of this disclosure.

Claims

1. Use of an effective amount of an aryl hydrocarbon receptor antagonist in the preparation of a medicament for treating cancer in patients in need, wherein the patient's benefit from treatment with the aryl hydrocarbon receptor antagonist is determined by the following steps: (a) Measuring or having measured the amount of at least one aryl hydrocarbon receptor ligand from a fecal sample obtained from the patient, wherein the at least one aryl hydrocarbon receptor ligand comprises kynurenine, tryptophan, phenylpyruvic acid, indolelacic acid, or a combination thereof. (b) The amount of at least one aryl hydrocarbon acceptor ligand has been compared with the threshold fraction, and (c) If the amount of the at least one aryl hydrocarbon receptor ligand is higher than the threshold fraction, it is determined, or has been determined, that the patient will benefit from treatment with an aryl hydrocarbon receptor antagonist, wherein the aryl hydrocarbon receptor antagonist is a compound of formula Ia: (Ia) Or its pharmaceutically acceptable salt. in: Ring A is selected from 6-10-membered aryl, 5-10-membered heteroaryl, 3-10-membered cycloalkyl, and 3-10-membered heterocycloalkyl, wherein each 6-10-membered aryl, 5-10-membered heteroaryl, 3-10-membered cycloalkyl, and 3-10-membered heterocycloalkyl is independently and optionally surrounded by 1 to 5 R. A Instance replacement; Ring B is selected from 6-10-membered aryl, 5-10-membered heteroaryl, 3-10-membered cycloalkyl, and 3-10-membered heterocycloalkyl, wherein each 6-10-membered aryl, 5-10-membered heteroaryl, 3-10-membered cycloalkyl, and 3-10-membered heterocycloalkyl is independently and optionally surrounded by 1 to 5 R. B Instance replacement; R is selected from hydrogen, C1-C 10 Alkyl, 6-10 aryl, -C(O)R', -C(O)NR'R', 3-10 cycloalkyl, -C(O)OR', C1-C 10 Heteroalkyl, 5-10-membered heteroaryl, 3-10-membered heterocyclic alkyl, amino, cyano, halogenated, hydroxyl, and -C(O)H, wherein each C1-C 10 Alkyl, 6-10 aryl, 3-10 cycloalkyl, C1-C 10 Heteroalkyl, 5-10-membered heteroaryl and 3-10-membered heterocyclic alkyl are independently and optionally bound by 1 to 5 Rs. C Instance replacement; Each R' is independently selected from hydrogen, C1-C 10 Alkyl, C1-C 10 Haloalkyl, C1-C 10 Hydroxyalkyl and C1-C 10 Heteroalkyl; Each R A Independently selected from halogenated, hydroxyl-containing, C1-C 10 Alkyl, C1-C 10 Haloalkyl, C1-C 10 Alkoxy, C1-C 10 Halogenated alkoxy groups, C1-C 10 Hydroxyalkyl and NR''R''; Each R B Independently selected from halogenated, hydroxyl-containing, C1-C 10 Alkyl, C1-C 10 Haloalkyl, C1-C 10 Alkoxy, C1-C 10 Halogenated alkoxy groups, C1-C 10 Hydroxyalkyl and NR''R''; Each R C Independently selected from halogenated, hydroxylated, cyano-based, C1-C 10 Alkyl, C1-C 10 Alkoxy, C1-C 10 Halogenated alkyl groups, 3-10 membered cycloalkyl groups, 3-10 membered heterocycloalkyl groups, 6-10 membered aryl groups, and 5-10 membered heteroaryl groups; and Each R'' is independently selected from hydrogen, C1-C 10 Alkyl, C1-C 10 Haloalkyl, C1-C 10 Hydroxyalkyl and C1-C 10 Heteroalkyl groups.

2. The use according to claim 1, wherein, The patient has esophageal cancer, multiple myeloma, melanoma, Hodgkin's lymphoma, lung cancer, head and neck cancer, mesothelioma, gastric cancer, urothelial carcinoma, renal cell carcinoma, ovarian cancer, fallopian tube cancer, peritoneal cancer, biliary tract cancer, breast cancer, pancreatic cancer, glioblastoma, prostate cancer, colon cancer, or liver cancer.

3. The use according to claim 1, wherein, The patient has non-small cell lung cancer, urothelial carcinoma, gastric and gastroesophageal junction cancer, head and neck squamous cell carcinoma, non-Hodgkin lymphoma, pleural mesothelioma, gastric adenocarcinoma, metastatic melanoma, renal cell carcinoma, bladder cancer, esophageal squamous cell carcinoma, triple-negative breast cancer, or prostate cancer.

4. The use according to claim 1, wherein, The patient has small cell lung cancer, diffuse large B-cell lymphoma, squamous cell carcinoma of the lung, transitional cell carcinoma, recurrent or metastatic PD-L1 positive or negative squamous cell carcinoma of the head and neck, or unresectable or metastatic melanoma.

5. The use according to claim 1, wherein, The patient has nasopharyngeal carcinoma, colorectal cancer, hepatocellular carcinoma, metastatic clear cell renal cell carcinoma, or metastatic noncutaneous melanoma.

6. The use according to any one of claims 1-5, wherein, The patient had been treated with immune checkpoint inhibitor therapy, but did not respond to it.

7. The use according to any one of claims 1-5, wherein, The patient had been treated with immune checkpoint inhibitor therapy and had developed resistance to it.

8. The use according to any one of claims 1-5, wherein, The at least one aryl hydrocarbon acceptor ligand comprises kynurenine, tryptophan, phenylpyruvic acid, and indolelactic acid.

9. The use according to any one of claims 1-5, wherein, The amount of at least one aryl hydrocarbon acceptor ligand in the sample was measured by liquid chromatography-mass spectrometry or gas chromatography-mass spectrometry.

10. The use according to any one of claims 1-5, wherein, The threshold score is determined by measuring the amount of at least one corresponding aryl hydrocarbon receptor ligand in a control stool sample from a patient who has received an immune checkpoint inhibitor therapy.

11. The use according to any one of claims 1-5, wherein: Cycloyl A is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, phenyl, pyrrolyl, furanyl, furazanyl, thienyl, imidazolyl, isothiazolyl, isoxazolyl, oxazolyl, oxadiazolyl, tetrazolyl, thiazolyl, triazolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyrrolylalkyl, pyrazolylalkyl, piperidinyl, piperazinyl, morpholino, azahexayl, tetrahydropyranyl, and tetrahydrofuranyl, wherein cyclopropyl, Each of the following groups is independently and optionally influenced by 1 to 5 R groups: cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, phenyl, pyrrolyl, furanyl, furazanyl, thienyl, imidazolyl, isothiazolyl, isoxazolyl, oxazolyl, oxadiazolyl, tetrazolyl, thiazolyl, triazolyl, pyrazolyl, pyridinyl, pyridine, pyrrolylalkyl, pyrazolylalkyl, piperidinyl, piperazinyl, morpholino, azahexyl, tetrahydropyranyl, and tetrahydrofuranyl. A Instance replacement; Ring B is selected from phenyl, benzodioxacyclopentenyl, pyrrolyl, furanyl, furazanyl, thiophene, imidazolyl, isothiazolyl, isoxazolyl, oxazolyl, oxadiazolyl, tetrazolyl, thiazolyl, triazolyl, pyrazolyl, pyridinyl, pyrazinyl, pyridinyl, pyridoneyl, and pyrimidinyl, wherein each of the following groups is independently and optionally selected by 1 to 5 R groups. B Instance replacement; Or ring B is R is selected from hydrogen, C1-C 10 Alkyl, 6-10 aryl, 3-10 cycloalkyl, 5-10 heteroaryl, and 3-10 heterocycloalkyl, wherein C1-C 10 Each of the following groups—alkyl, 6-10 aryl, 3-10 cycloalkyl, 5-10 heteroaryl, and 3-10 heterocycloalkyl—is independently and optionally bounded by 1 to 5 R groups. C Instance replacement; Each R A Independently selected from halogenated, C1-C 10 Alkyl, C1-C 10 Halogenated alkyl groups and C1-C 10 Halogenated alkoxy groups; Each R B Independently selected from halogenated, C1-C 10 Alkyl and C1-C 10 Halogenated alkyl groups; Each R C Independently selected from halogenated, hydroxyl-containing, and C1-C 10 Alkyl group.

12. The use according to any one of claims 1-5, wherein: Ring A is selected from pyrrolidinyl, pyrazolyl, piperidinyl, piperazinyl, morpholino, azahexayl, tetrahydropyranyl, and tetrahydrofuranyl. Each of the following groups—pyrrolidinyl, pyrazolyl, piperidinyl, piperazinyl, morpholino, aziridine, tetrahydropyranyl, and tetrahydrofuranyl—is independently and optionally influenced by one to three R groups. A Instance replacement; Ring B is selected from benzodioxanepentenyl, pyrroleyl, furanyl, furazanyl, thienyl, imidazolyl, isothiazolyl, isoxazolyl, oxazolyl, oxadiazolyl, tetrazolyl, thiazolyl, triazolyl, pyrazolyl, pyridinyl, pyrazinyl, pyridinyl, pyridoneyl, and pyrimidinyl. Each of the following groups—benzodioxanepentenyl, pyrroleyl, furanyl, furazanyl, thienyl, imidazolyl, isothiazolyl, isoxazolyl, oxazolyl, oxadiazolyl, tetrazolyl, thiazolyl, triazolyl, pyrazolyl, pyridyl, pyrazinyl, pyridazinyl, and pyrimidinyl—is independently and optionally influenced by one to three R groups. B The instance is replaced.

13. The use according to any one of claims 1-5, wherein, The aryl hydrocarbon receptor antagonist is any one of the following compounds, or a pharmaceutically acceptable salt thereof: 。 14. Use of an immune checkpoint inhibitor and an aryl hydrocarbon receptor antagonist in the preparation of a medicament for treating cancer in a patient in need, wherein the patient who would benefit from a combination therapy comprising the immune checkpoint inhibitor and the aryl hydrocarbon receptor is determined by the following steps: (a) Measuring or having measured the amount of at least one aryl hydrocarbon receptor ligand from a fecal sample obtained from the patient, wherein the at least one aryl hydrocarbon receptor ligand comprises kynurenine, tryptophan, phenylpyruvic acid, indolelacic acid, or a combination thereof. (b) The amount of at least one aryl hydrocarbon acceptor ligand has been compared with the threshold fraction, and (c) If the amount of the at least one aryl hydrocarbon receptor ligand is higher than the threshold fraction, then it is determined, or has been determined, that the patient will benefit from the combination therapy. The aryl hydrocarbon receptor antagonist is a compound of formula Ia: (Ia) Or its pharmaceutically acceptable salt. in: Ring A is selected from 6-10-membered aryl, 5-10-membered heteroaryl, 3-10-membered cycloalkyl, and 3-10-membered heterocycloalkyl, wherein each 6-10-membered aryl, 5-10-membered heteroaryl, 3-10-membered cycloalkyl, and 3-10-membered heterocycloalkyl is independently and optionally surrounded by 1 to 5 R. A Instance replacement; Ring B is selected from 6-10-membered aryl, 5-10-membered heteroaryl, 3-10-membered cycloalkyl, and 3-10-membered heterocycloalkyl, wherein each 6-10-membered aryl, 5-10-membered heteroaryl, 3-10-membered cycloalkyl, and 3-10-membered heterocycloalkyl is independently and optionally surrounded by 1 to 5 R. B Instance replacement; R is selected from hydrogen, C1-C 10 Alkyl, 6-10 aryl, -C(O)R', -C(O)NR'R', 3-10 cycloalkyl, -C(O)OR', C1-C 10 Heteroalkyl, 5-10-membered heteroaryl, 3-10-membered heterocyclic alkyl, amino, cyano, halogenated, hydroxyl, and -C(O)H, wherein each C1-C 10 Alkyl, 6-10 aryl, 3-10 cycloalkyl, C1-C 10 Heteroalkyl, 5-10-membered heteroaryl and 3-10-membered heterocyclic alkyl are independently and optionally bound by 1 to 5 Rs. C Instance replacement; Each R' is independently selected from hydrogen, C1-C 10 Alkyl, C1-C 10 Haloalkyl, C1-C 10 Hydroxyalkyl and C1-C 10 Heteroalkyl; Each R A Independently selected from halogenated, hydroxyl-containing, C1-C 10 Alkyl, C1-C 10 Haloalkyl, C1-C 10 Alkoxy, C1-C 10 Halogenated alkoxy groups, C1-C 10 Hydroxyalkyl and NR''R''; Each R B Independently selected from halogenated, hydroxyl-containing, C1-C 10 Alkyl, C1-C 10 Haloalkyl, C1-C 10 Alkoxy, C1-C 10 Halogenated alkoxy groups, C1-C 10 Hydroxyalkyl and NR''R''; Each R C Independently selected from halogenated, hydroxylated, cyano-based, C1-C 10 Alkyl, C1-C 10 Alkoxy, C1-C 10 Halogenated alkyl groups, 3-10 membered cycloalkyl groups, 3-10 membered heterocycloalkyl groups, 6-10 membered aryl groups, and 5-10 membered heteroaryl groups; and Each R'' is independently selected from hydrogen, C1-C 10 Alkyl, C1-C 10 Haloalkyl, C1-C 10 Hydroxyalkyl and C1-C 10 Heteroalkyl groups.

15. The use according to claim 14, wherein, The patient has esophageal cancer, multiple myeloma, melanoma, Hodgkin's lymphoma, lung cancer, head and neck cancer, mesothelioma, gastric cancer, urothelial carcinoma, renal cell carcinoma, ovarian cancer, fallopian tube cancer, peritoneal cancer, biliary tract cancer, breast cancer, pancreatic cancer, glioblastoma, prostate cancer, colon cancer, or liver cancer.

16. The use according to claim 14, wherein, The patient has non-small cell lung cancer, urothelial carcinoma, gastric and gastroesophageal junction cancer, head and neck squamous cell carcinoma, non-Hodgkin lymphoma, pleural mesothelioma, gastric adenocarcinoma, metastatic melanoma, renal cell carcinoma, bladder cancer, esophageal squamous cell carcinoma, triple-negative breast cancer, or prostate cancer.

17. The use according to claim 14, wherein, The patient has small cell lung cancer, diffuse large B-cell lymphoma, squamous cell carcinoma of the lung, transitional cell carcinoma, recurrent or metastatic PD-L1 positive or negative squamous cell carcinoma of the head and neck, or unresectable or metastatic melanoma.

18. The use according to claim 14, wherein, The patient has nasopharyngeal carcinoma, colorectal cancer, hepatocellular carcinoma, metastatic clear cell renal cell carcinoma, or metastatic noncutaneous melanoma.

19. The use according to any one of claims 14-18, wherein, The patient had been treated with immune checkpoint inhibitor therapy but did not respond to it.

20. The use according to any one of claims 14-18, wherein, The patient had been treated with immune checkpoint inhibitor therapy and had developed resistance to it.

21. The use according to any one of claims 14-18, wherein, The at least one aryl hydrocarbon acceptor ligand comprises kynurenine, tryptophan, phenylpyruvic acid, and indolelactic acid.

22. The use according to any one of claims 14-18, wherein, The amount of at least one aryl hydrocarbon acceptor ligand in the sample was measured by liquid chromatography-mass spectrometry or gas chromatography-mass spectrometry.

23. The use according to any one of claims 14-18, wherein, This immune checkpoint inhibitor therapy includes anti-PD-1 antibodies, anti-PD-L1 antibodies, anti-CTLA-4 antibodies, anti-LAG-3 antibodies, anti-TIM-3 antibodies, anti-TIGIT antibodies, anti-VISTA antibodies, or combinations thereof.

24. The use according to any one of claims 14-18, wherein, The threshold score is determined by measuring the amount of at least one corresponding aryl hydrocarbon receptor ligand in a control stool sample from a patient who has received an immune checkpoint inhibitor therapy.

25. The use according to any one of claims 14-18, wherein: Cycloyl A is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, phenyl, pyrrolyl, furanyl, furazanyl, thienyl, imidazolyl, isothiazolyl, isoxazolyl, oxazolyl, oxadiazolyl, tetrazolyl, thiazolyl, triazolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyrrolylalkyl, pyrazolylalkyl, piperidinyl, piperazinyl, morpholino, azahexayl, tetrahydropyranyl, and tetrahydrofuranyl, wherein cyclopropyl, Each of the following groups is independently and optionally influenced by 1 to 5 R groups: cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, phenyl, pyrrolyl, furanyl, furazanyl, thienyl, imidazolyl, isothiazolyl, isoxazolyl, oxazolyl, oxadiazolyl, tetrazolyl, thiazolyl, triazolyl, pyrazolyl, pyridinyl, pyridine, pyrrolylalkyl, pyrazolylalkyl, piperidinyl, piperazinyl, morpholino, azahexyl, tetrahydropyranyl, and tetrahydrofuranyl. A Instance replacement; Ring B is selected from phenyl, benzodioxacyclopentenyl, pyrrolyl, furanyl, furazanyl, thiophene, imidazolyl, isothiazolyl, isoxazolyl, oxazolyl, oxadiazolyl, tetrazolyl, thiazolyl, triazolyl, pyrazolyl, pyridinyl, pyrazinyl, pyridinyl, pyridoneyl, and pyrimidinyl, wherein each of the following groups is independently and optionally selected by 1 to 5 R groups. B Instance replacement; Or ring B is R is selected from hydrogen, C1-C 10 Alkyl, 6-10 aryl, 3-10 cycloalkyl, 5-10 heteroaryl, and 3-10 heterocycloalkyl, wherein C1-C 10 Each of the following groups—alkyl, 6-10 aryl, 3-10 cycloalkyl, 5-10 heteroaryl, and 3-10 heterocycloalkyl—is independently and optionally bounded by 1 to 5 R groups. C Instance replacement; Each R A Independently selected from halogenated, C1-C 10 Alkyl, C1-C 10 Halogenated alkyl groups and C1-C 10 Halogenated alkoxy groups; Each R B Independently selected from halogenated, C1-C 10 Alkyl and C1-C 10 Halogenated alkyl groups; Each R C Independently selected from halogenated, hydroxyl-containing, and C1-C 10 Alkyl group.

26. The use according to any one of claims 14-18, wherein: Ring A is selected from pyrrolidinyl, pyrazolyl, piperidinyl, piperazinyl, morpholino, azahexayl, tetrahydropyranyl, and tetrahydrofuranyl. Each of the following groups—pyrrolidinyl, pyrazolyl, piperidinyl, piperazinyl, morpholino, aziridine, tetrahydropyranyl, and tetrahydrofuranyl—is independently and optionally influenced by one to three R groups. A Instance replacement; Ring B is selected from benzodioxanepentenyl, pyrroleyl, furanyl, furazanyl, thienyl, imidazolyl, isothiazolyl, isoxazolyl, oxazolyl, oxadiazolyl, tetrazolyl, thiazolyl, triazolyl, pyrazolyl, pyridinyl, pyrazinyl, pyridinyl, pyridoneyl, and pyrimidinyl. Each of the following groups—benzodioxanepentenyl, pyrroleyl, furanyl, furazanyl, thienyl, imidazolyl, isothiazolyl, isoxazolyl, oxazolyl, oxadiazolyl, tetrazolyl, thiazolyl, triazolyl, pyrazolyl, pyridyl, pyrazinyl, pyridazinyl, and pyrimidinyl—is independently and optionally influenced by one to three R groups. B The instance is replaced.

27. The use according to any one of claims 14-18, wherein, The aryl hydrocarbon receptor antagonist is any one of the following compounds, or a pharmaceutically acceptable salt thereof: 。 28. The use of immune checkpoint inhibitors in the preparation of drugs for treating patients with cancer, wherein, The following steps determine whether the patient's cancer is responsive to immune checkpoint inhibitor therapy: (a) Measuring or having measured the amount of at least one aryl hydrocarbon receptor ligand from a fecal sample obtained from the patient, wherein the at least one aryl hydrocarbon receptor ligand comprises kynurenine, tryptophan, phenylpyruvic acid, indolelacic acid, or a combination thereof. (b) The amount of at least one aryl hydrocarbon acceptor ligand has been compared with the threshold fraction, and (c) If the amount of the at least one aryl hydrocarbon receptor ligand is equal to or less than the threshold fraction, then the patient’s cancer is determined to be responsive to immune checkpoint inhibitor therapy.

29. The use according to claim 28, wherein, The patient has esophageal cancer, multiple myeloma, melanoma, Hodgkin's lymphoma, lung cancer, head and neck cancer, mesothelioma, gastric cancer, renal cell carcinoma, ovarian cancer, fallopian tube cancer, peritoneal cancer, biliary tract cancer, breast cancer, pancreatic cancer, glioblastoma, prostate cancer, colon cancer, or liver cancer.

30. The use according to claim 28, wherein, The patient has non-small cell lung cancer, urothelial carcinoma, gastric and gastroesophageal junction cancer, head and neck squamous cell carcinoma, non-Hodgkin lymphoma, pleural mesothelioma, gastric adenocarcinoma, metastatic melanoma, renal cell carcinoma, bladder cancer, esophageal squamous cell carcinoma, triple-negative breast cancer, or prostate cancer.

31. The use according to claim 28, wherein, The patient has small cell lung cancer, diffuse large B-cell lymphoma, squamous cell carcinoma of the lung, transitional cell carcinoma, recurrent or metastatic PD-L1 positive or negative squamous cell carcinoma of the head and neck, or unresectable or metastatic melanoma.

32. The use according to claim 28, wherein, The patient has nasopharyngeal carcinoma, colorectal cancer, hepatocellular carcinoma, metastatic clear cell renal cell carcinoma, or metastatic noncutaneous melanoma.

33. The use according to any one of claims 28-32, wherein, The at least one aryl hydrocarbon acceptor ligand comprises kynurenine, tryptophan, phenylpyruvic acid, and indolelactic acid.

34. The use according to any one of claims 28-32, wherein, The amount of at least one aryl hydrocarbon acceptor ligand in the sample was measured by liquid chromatography-mass spectrometry or gas chromatography-mass spectrometry.

35. The use according to any one of claims 28-32, wherein, This immune checkpoint inhibitor therapy includes anti-PD-1 antibodies, anti-PD-L1 antibodies, anti-CTLA-4 antibodies, anti-LAG-3 antibodies, anti-TIM-3 antibodies, anti-TIGIT antibodies, anti-VISTA antibodies, or combinations thereof.

36. The use according to any one of claims 28-32, wherein, The threshold score is determined by measuring the amount of at least one corresponding aryl hydrocarbon receptor ligand in a control stool sample from a patient who has received an immune checkpoint inhibitor therapy.

37. The use according to any one of claims 28-32, wherein, If the amount of at least one aryl hydrocarbon receptor ligand is higher than the threshold fraction, it is determined that the patient's cancer is responsive to immune checkpoint inhibitor therapy in combination with an aryl hydrocarbon receptor antagonist. The aryl hydrocarbon receptor antagonist is a compound of formula Ia: (Ia) Or its pharmaceutically acceptable salt. in: Ring A is selected from 6-10-membered aryl, 5-10-membered heteroaryl, 3-10-membered cycloalkyl, and 3-10-membered heterocycloalkyl, wherein each 6-10-membered aryl, 5-10-membered heteroaryl, 3-10-membered cycloalkyl, and 3-10-membered heterocycloalkyl is independently and optionally surrounded by 1 to 5 R. A Instance replacement; Ring B is selected from 6-10-membered aryl, 5-10-membered heteroaryl, 3-10-membered cycloalkyl, and 3-10-membered heterocycloalkyl, wherein each 6-10-membered aryl, 5-10-membered heteroaryl, 3-10-membered cycloalkyl, and 3-10-membered heterocycloalkyl is independently and optionally surrounded by 1 to 5 R. B Instance replacement; R is selected from hydrogen, C1-C 10 Alkyl, 6-10 aryl, -C(O)R', -C(O)NR'R', 3-10 cycloalkyl, -C(O)OR', C1-C 10 Heteroalkyl, 5-10-membered heteroaryl, 3-10-membered heterocyclic alkyl, amino, cyano, halogenated, hydroxyl, and -C(O)H, wherein each C1-C 10 Alkyl, 6-10 aryl, 3-10 cycloalkyl, C1-C 10 Heteroalkyl, 5-10-membered heteroaryl and 3-10-membered heterocyclic alkyl are independently and optionally bound by 1 to 5 Rs. C Instance replacement; Each R' is independently selected from hydrogen, C1-C 10 Alkyl, C1-C 10 Haloalkyl, C1-C 10 Hydroxyalkyl and C1-C 10 Heteroalkyl; Each R A Independently selected from halogenated, hydroxyl-containing, C1-C 10 Alkyl, C1-C 10 Haloalkyl, C1-C 10 Alkoxy, C1-C 10 Halogenated alkoxy groups, C1-C 10 Hydroxyalkyl and NR''R''; Each R B Independently selected from halogenated, hydroxyl-containing, C1-C 10 Alkyl, C1-C 10 Haloalkyl, C1-C 10 Alkoxy, C1-C 10 Halogenated alkoxy groups, C1-C 10 Hydroxyalkyl and NR''R''; Each R C Independently selected from halogenated, hydroxylated, cyano-based, C1-C 10 Alkyl, C1-C 10 Alkoxy, C1-C 10 Halogenated alkyl groups, 3-10 membered cycloalkyl groups, 3-10 membered heterocycloalkyl groups, 6-10 membered aryl groups, and 5-10 membered heteroaryl groups; and Each R'' is independently selected from hydrogen, C1-C 10 Alkyl, C1-C 10 Haloalkyl, C1-C 10 Hydroxyalkyl and C1-C 10 Heteroalkyl groups.

38. The use according to any one of claims 28-32, wherein: Cycloyl A is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, phenyl, pyrrolyl, furanyl, furazanyl, thienyl, imidazolyl, isothiazolyl, isoxazolyl, oxazolyl, oxadiazolyl, tetrazolyl, thiazolyl, triazolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyrrolylalkyl, pyrazolylalkyl, piperidinyl, piperazinyl, morpholino, azahexayl, tetrahydropyranyl, and tetrahydrofuranyl, wherein cyclopropyl, Each of the following groups is independently and optionally influenced by 1 to 5 R groups: cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, phenyl, pyrrolyl, furanyl, furazanyl, thienyl, imidazolyl, isothiazolyl, isoxazolyl, oxazolyl, oxadiazolyl, tetrazolyl, thiazolyl, triazolyl, pyrazolyl, pyridinyl, pyridine, pyrrolylalkyl, pyrazolylalkyl, piperidinyl, piperazinyl, morpholino, azahexyl, tetrahydropyranyl, and tetrahydrofuranyl. A Instance replacement; Ring B is selected from phenyl, benzodioxacyclopentenyl, pyrrolyl, furanyl, furazanyl, thiophene, imidazolyl, isothiazolyl, isoxazolyl, oxazolyl, oxadiazolyl, tetrazolyl, thiazolyl, triazolyl, pyrazolyl, pyridinyl, pyrazinyl, pyridinyl, pyridoneyl, and pyrimidinyl, wherein each of the following groups is independently and optionally selected by 1 to 5 R groups. B Instance replacement; Or ring B is R is selected from hydrogen, C1-C 10 Alkyl, 6-10 aryl, 3-10 cycloalkyl, 5-10 heteroaryl, and 3-10 heterocycloalkyl, wherein C1-C 10 Each of the following groups—alkyl, 6-10 aryl, 3-10 cycloalkyl, 5-10 heteroaryl, and 3-10 heterocycloalkyl—is independently and optionally bounded by 1 to 5 R groups. C Instance replacement; Each R A Independently selected from halogenated, C1-C 10 Alkyl, C1-C 10 Halogenated alkyl groups and C1-C 10 Halogenated alkoxy groups; Each R B Independently selected from halogenated, C1-C 10 Alkyl and C1-C 10 Halogenated alkyl groups; Each R C Independently selected from halogenated, hydroxyl-containing, and C1-C 10 Alkyl group.

39. The use according to any one of claims 28-32, wherein: Ring A is selected from pyrrolidinyl, pyrazolyl, piperidinyl, piperazinyl, morpholino, azahexayl, tetrahydropyranyl, and tetrahydrofuranyl. Each of the following groups—pyrrolidinyl, pyrazolyl, piperidinyl, piperazinyl, morpholino, aziridine, tetrahydropyranyl, and tetrahydrofuranyl—is independently and optionally influenced by one to three R groups. A Instance replacement; Ring B is selected from benzodioxanepentenyl, pyrroleyl, furanyl, furazanyl, thienyl, imidazolyl, isothiazolyl, isoxazolyl, oxazolyl, oxadiazolyl, tetrazolyl, thiazolyl, triazolyl, pyrazolyl, pyridinyl, pyrazinyl, pyridinyl, pyridoneyl, and pyrimidinyl. Each of the following groups—benzodioxanepentenyl, pyrroleyl, furanyl, furazanyl, thienyl, imidazolyl, isothiazolyl, isoxazolyl, oxazolyl, oxadiazolyl, tetrazolyl, thiazolyl, triazolyl, pyrazolyl, pyridyl, pyrazinyl, pyridazinyl, and pyrimidinyl—is independently and optionally influenced by one to three R groups. B The instance is replaced.

40. The use according to any one of claims 28-32, wherein, The aryl hydrocarbon receptor antagonist is any one of the following compounds, or a pharmaceutically acceptable salt thereof: 。 41. The use according to any one of claims 1-5 or 14-18, wherein the aryl hydrocarbon receptor antagonist is any one of the following compounds or a pharmaceutically acceptable salt thereof: (3-(2-hydroxy-2-methylpropyl)-8-(pyridin-3-yl)-6-(6-trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one); (6-(4-chlorophenyl)-3-(2-hydroxy-2-methylpropyl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one); (3-(2-hydroxy-2-methylpropyl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(6-trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one); (3-((3R,4S)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)-6-(6-trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one); (3-(2-hydroxy-2-methylpropyl)-8-(pyridin-3-yl)-6-(5-trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one); or (6-(4-chlorophenyl)-3-((3S,4R)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one).

42. The use according to claim 37, wherein the aryl hydrocarbon receptor antagonist is any one of the following compounds or a pharmaceutically acceptable salt thereof: (3-(2-hydroxy-2-methylpropyl)-8-(pyridin-3-yl)-6-(6-trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one); (6-(4-chlorophenyl)-3-(2-hydroxy-2-methylpropyl)-8-(pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one); (3-(2-hydroxy-2-methylpropyl)-8-(1-methyl-1H-pyrazol-4-yl)-6-(6-trifluoromethyl)pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one); (3-((3R,4S)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)-6-(6-trifluoromethyl)pyridin-3-yl)pyrido[3,4-d]pyrimidin-4(3H)-one); (3-(2-hydroxy-2-methylpropyl)-8-(pyridin-3-yl)-6-(5-trifluoromethyl)pyridin-2-yl)pyrido[3,4-d]pyrimidin-4(3H)-one); or (6-(4-chlorophenyl)-3-((3S,4R)-4-hydroxytetrahydrofuran-3-yl)-8-(pyridin-3-yl)pyridino[3,4-d]pyrimidin-4(3H)-one).

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