Methods and compositions for treating cancer

Abstract

The present disclosure is directed to compounds, compositions, and methods for the treatment of various diseases and / or conditions associated with G protein-coupled receptor 174 (eg, cancer). TIFF2022517978000110.tif171148

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Patent Application No. 62 / 791,591, filed January 11, 2019, U.S. Provisional Patent Application No. 62 / 886,235, filed August 13, 2019, U.S. Provisional Patent Application No. 62 / 936,223, filed November 15, 2019, and U.S. Provisional Patent Application No. 62 / 946,631, filed December 11, 2019. Each of the foregoing related applications is incorporated herein by reference in its entirety.

[0002] Sequence Listing Description The Sequence Listing associated with this application is provided in text format in lieu of a paper copy and is incorporated herein by reference. The text file containing the Sequence Listing is named OG_1_0294_US_Sequence_Listing_20200107_ST25.txt. The file is 11 KB, was created on January 7, 2020, and has been submitted via EFS-Web in conjunction with the filing of this application.

[0003] Technical Field The present disclosure is directed to methods and compounds for treating cancer by administering to a subject a therapeutically effective amount of an inhibitor of G protein-coupled receptor 174 (GPR174)-mediated signaling, or a combination of an inhibitor of GPR174-mediated signaling with an inhibitor of ATP-adenosine A2aR or A2bR-mediated signaling (such as an A2aR antagonist and / or an A2bR antagonist), or a combination of an inhibitor of GPR174-mediated signaling with an inhibitor of enzymatic pathways involved in the production of adenosine, e.g., CD38, CD39, and / or CD73 (such as a CD73 inhibitor and / or a CD38 inhibitor and / or a CD39 inhibitor), thereby stimulating an immune response in a subject in need thereof, e.g., a subject suffering from cancer. [Background technology]

[0004] background Mammalian G protein-coupled receptors (GPCRs) constitute a diverse superfamily of proteins with hundreds of members. GPCRs act as receptors for numerous different signals. Sensory GPCRs (spurs) are receptors for sensory signals of external origin, perceived as different colors of light, odors, pheromones, or tastes. Most other GPCRs respond to endogenous signals, such as peptides, lipids, neurotransmitters, or nucleotides. GPCRs belonging to the latter group are involved in numerous physiological processes, including the control of neuronal excitability, metabolism, reproduction, development, cell division, hormone homeostasis, and behavior, and are differentially expressed in many cell types within the body.

[0005] Ligands have been identified for several GPCRs. GPCRs that have yet to have identified endogenous or cognate ligands are known as orphan GPCRs. As of 2019, approximately 120 GPCRs, including GPR174, were considered orphan GPCRs (see "Orphan and other 7TM receptors, IUPHAR / BPS Guide to Pharmacology," accessed August 6, 2019; Alexander S. et al., Br. J. Pharmacol. 174 Suppl 1:S17-S129, 2017). Of all currently available drugs, 30-40% are modulators of specific GPCRs. Of the approximately 240 non-orphan GPCRs, more than half are targeted by marketed drugs or drugs currently in clinical development (see Hauser AS et al., Nature Reviews Drug Discovery 16(12):829-842, 2017).

[0006] The development of drugs targeting specific GPCRs generally requires knowledge of endogenous or surrogate ligands, i.e., non-endogenous molecules that bind to and functionally interact with a specific GPCR. This knowledge can serve as a screening control and / or as a medicinal chemistry basis for developing synthetic drug molecules that functionally interact with that specific GPCR. Given the potential use of orphan GPCRs as targets for new drugs, the identification of ligands or surrogate ligands for orphan GPCRs is believed to facilitate the development of new drugs against those receptors. However, despite extensive efforts to identify such ligands, a large number of orphan receptors remain (see, e.g., Levoye et al., Drug Discov. Today 13(1-2):52-8, 2008). Conventional GPCR screening strategies typically focus on the detection of Ca2+ in response to GPCR activation. ++ Drug discovery relies on screening using both biochemical and functional assays, such as assays that measure signal transduction. However, these traditional approaches rely on the presence of endogenous or surrogate ligands. When used with orphan GPCRs, these assays can only identify agonists for the GPCR. Traditional assay techniques cannot identify inverse agonists or antagonists until an agonist is available. Importantly, many FDA-approved drugs targeting GPCRs are functional inhibitors (e.g., partial agonists, inverse agonists, partial inverse agonists, or antagonists). Therefore, traditional approaches fail to detect this common and important class of GPCR ligands.

[0007] GPR174, also known as FKSG79 and GPCR17, is expressed in a relatively small number of tissues, including the spleen, thymus, bone marrow, and lymph nodes (Regard et al., Cell 135:561-71, 2008 (Non-Patent Document 5); Chu et al., J Med Genet, 50:479-85, 2013 (Non-Patent Document 6); Sugita et al., Biochem Biophys Res Commun 430:190-5, 2013 (Non-Patent Document 7)). Within lymphoid tissues, GPR174 is expressed at high levels in naive B and T cells, particularly in regulatory T cells (Tregs) (Barnes et al., J Exp Med 212:1011-20, 2015 (Non-Patent Document 8)).

[0008] As described herein, GPR174 activity is related to cancer.In view of the role that GPR174 seems to play in cancer, it is desirable to identify the inhibitor of this receptor.There is a need in the art for effective cancer treatment.The methods and compositions provided herein address these and other deficiencies in the art. [Prior art documents] [Non-patent literature]

[0009] [Non-Patent Document 1] Orphan and other 7TM receptors, IUPHAR / BPS Guide to Pharmacology (accessed August 6, 2019) [Non-patent document 2] Alexander S. et al., Br. J. Pharmacol. 174 Suppl 1:S17-S129,2017 [Non-patent document 3] Hauser AS et al., Nature Reviews Drug Discovery 16(12):829-842,2017 [Non-patent document 4] Levoye et al., Drug Discov. Today 13(1-2):52-8, 2008 [Non-Patent Document 5] Regard et al., Cell 135:561-71, 2008 [Non-patent document 6] Chu et al., J Med Genet, 50:479-85, 2013 [Non-Patent Document 7] Sugita et al., Biochem Biophys Res Commun 430:190-5,2013 [Non-patent document 8] Barnes et al., J Exp Med 212:1011-20,2015 Summary of the Invention

[0010] Quick Overview The present inventors have identified chemical compounds that functionally interact with GPR174 and inhibit one or more GPR174-mediated signaling pathways, and further characterized the GPR174 receptor signaling profile, including the Gs signaling pathway. The present inventors demonstrated that representative GPR174 inhibitory compounds can reduce the fraction of highly suppressive regulatory T cells or "T-Regs" (FoxP3+Helios+) in human peripheral blood mononuclear cells (PBMCs), as described in Examples 6, 8, and 9. The present inventors further demonstrated that GPR174 inhibition stimulates IL-2 production in PBMCs in a dose-dependent manner, as described in Examples 6-11. The present inventors have also demonstrated that GPR174 inhibition reduces immune cell-associated programmed death-ligand 1 (PD-L1) expression, cytotoxic T-lymphocyte-associated antigen 4 (CTLA4) expression, T cell immunoreceptor with Ig and ITIM domains (TIGIT) expression, and amphiregulin (AREG) expression, as described in Examples 13 and 14. Thus, GPR174-inhibitory compounds can be used as drugs to stimulate immune responses in mammalian subjects and can form the basis for further therapeutic agents. Based on these findings, the present disclosure relates to in vivo and in vitro methods for inhibiting the GPR174 receptor and thereby stimulating immune responses, particularly in subjects suffering from non-malignant or malignant neoplasms (i.e., cancer).

[0011] As further described herein, the inventors have also demonstrated that combining inhibition of GPR174 with inhibition of the adenosine 2a receptor (A2aR) and / or inhibition of the adenosine 2b receptor (A2bR), as described in Examples 15, 18-23, 27, and 29, results in synergistic induction of Th1 cytokine production (e.g., IFN-γ, IL-2, TNF, and GM-CSF) in human PBMCs.

[0012] As further described herein, the inventors also demonstrated that mouse colon cancer tumor growth and melanoma tumor growth were reduced in GPR174-deficient mice treated with anti-GITR antibody (i.e., a Treg-attenuating agent) compared to wild-type mice, as described in Examples 24 and 25.

[0013] As further described herein, the inventors have demonstrated that cancer cell-derived exosomes stimulate GPR174, and that inhibition of GPR174 in the presence of cancer cell-derived exosomes enhances Th1 cytokine levels, thereby stimulating and / or amplifying the immune system, as described in Example 26.

[0014] In summary, as demonstrated herein, GPR174 is an immune system-restricted Gαs-coupled GPCR, and PS exposed on liposomes and cell membranes stimulates GPR174, supporting a model of active GPR174-mediated immunosuppression in the tumor microenvironment. In the presence of both PS / lysoPS and adenosine, inhibition of both axes is required for effective restoration of T cell function. As further demonstrated herein, GPR174 deficiency enhances antitumor immune responses in mice.

[0015] Thus, in one aspect, the present disclosure provides methods and compositions for stimulating and / or amplifying an immune response (e.g., to treat cancer) by administering a therapeutically effective amount of an inhibitor of GPR174-mediated signaling to a subject suffering from cancer or cancer metastasis, or a subject at risk of developing cancer or cancer metastasis, as a single agent or optionally in combination with an inhibitor of ATP-adenosine A2aR or A2bR-mediated signaling (such as an A2aR antagonist, an A2bR antagonist and / or a CD73 inhibitor and / or a CD38 inhibitor and / or a CD39 inhibitor, and / or a Treg attenuating agent, or a combination thereof), thereby stimulating and / or amplifying an immune response in a mammalian subject in need thereof. In some embodiments, the method comprises administering an inhibitor of GPR174 and an A2aR antagonist to a subject suffering from cancer. In some embodiments, the method comprises administering to a subject suffering from cancer an inhibitor of GPR174 and an A2bR antagonist. In some embodiments, the method comprises administering to a subject suffering from cancer an inhibitor of GPR174 and an A2aR / A2bR antagonist. In some embodiments, the method comprises administering to a subject suffering from cancer an inhibitor of GPR174 and a CD73 inhibitor. In some embodiments, the method comprises administering to a subject suffering from cancer an inhibitor of GPR174 and a CD38 inhibitor. In some embodiments, the method comprises administering to a subject suffering from cancer a GPR174 inhibitor and a CD39 inhibitor. In some embodiments, the method comprises administering to a subject suffering from cancer an inhibitor of GPR174 and a Treg-attenuating agent. In some embodiments, the method comprises administering to a subject suffering from cancer a GPR174 inhibitor and an A2aR antagonist, and optionally at least one or more of an A2bR antagonist, a CD73 inhibitor, a CD38 inhibitor, a CD39 inhibitor, and a Treg attenuating agent.In some embodiments, the A2aR antagonist is a small molecule.In some embodiments, the A2aR antagonist is an antibody.In some embodiments, the A2aR antagonist is a peptide or nucleotide / nucleic acid based molecule. In some embodiments, the A2bR antagonist is a small molecule. In some embodiments, the A2bR antagonist is an antibody. In some embodiments, the A2bR antagonist is a peptide or nucleotide / nucleic acid based molecule. In some embodiments, the CD38 inhibitor is a small molecule. In some embodiments, the CD38 inhibitor is an antibody. In some embodiments, the CD38 antagonist is a peptide or nucleotide / nucleic acid based molecule. In some embodiments, the CD39 inhibitor is a small molecule. In some embodiments, the CD39 inhibitor is an antibody. In some embodiments, the CD39 antagonist is a peptide or nucleotide / nucleic acid based molecule. In some embodiments, the CD73 inhibitor is a small molecule. In some embodiments, the CD73 inhibitor is an antibody. In some embodiments, the CD73 antagonist is a peptide or nucleotide / nucleic acid based molecule. In some embodiments, the Treg attenuating agent is a small molecule. In some embodiments, the Treg attenuating agent is an antibody. In some embodiments, the Treg attenuating agent is a peptide or a nucleotide / nucleic acid-based molecule. In some embodiments, the GPR174 inhibitor is a small molecule. In some embodiments, the GPR174 inhibitor is an antibody. In some embodiments of the method, the GPR174 inhibitor and at least one or more of the A2aR antagonist, A2bR antagonist, CD38 inhibitor, CD39 inhibitor, CD73 inhibitor, and Treg attenuating agent are administered simultaneously (e.g., co-administered separately or together) or sequentially in any order, provided that the effect of the first administered inhibitor is still present at the time of the second (and optionally third) administered inhibitor or attenuating agent.

[0016] As further described in Examples 16 and 17 herein, the inventors have also discovered that phosphatidylserine (PS) is an agonist of GPR174-mediated Gs signaling. Example 17 provides experimental results demonstrating that apoptotic cells stimulate the GPR174 Gs signaling pathway in cells expressing GPR174. As further described in Example 16 and shown in Figures 47A-47F, PS-mediated GPR174 Gs signaling is inhibited by representative GPR174 inhibitory compounds 6, 10, 11, 20, 23, and 30, which belong to different chemical classes (i.e., groups I, II, and IV). This data demonstrates that compounds 6, 10, 11, 20, 23, and 30 act as GPR174 antagonists and inhibit PS liposome-mediated cAMP signaling. Therefore, GPR174-mediated PS liposome signaling is inhibited by a plurality of GPR174 inhibitory small molecule compounds with diverse chemical structures. Thus, in one aspect, the present disclosure provides a method for inhibiting PS-mediated activation of GPR174 Gs signaling in GPR174-expressing immune cells, comprising contacting the immune cells with an inhibitor of GPR174 Gs signaling, either as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent. In one aspect, the present disclosure provides a method of stimulating T cell-mediated immunity in a subject suffering from cancer, comprising administering to the subject a GPR174 inhibitor, either as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg-attenuating agent, in an amount effective to stimulate T cell-mediated immunity as described herein.

[0017] In another aspect, the present disclosure provides a method for increasing an anti-tumor immune response in a subject currently undergoing or having undergone treatment with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, comprising administering to the subject an effective amount of a GPR174 inhibitor to stimulate an enhanced anti-tumor immune response, although due to the fact that it may be preferable to keep CD73 active during initial tumor formation or in the setting of metastasis, the use of a CD73 inhibitor is preferably avoided in such situations.

[0018] In some embodiments, provided herein are methods of stimulating an immune response in a mammalian patient in need thereof, comprising administering to the mammalian patient a small molecule inhibitor of GPR174 signaling, alone or in combination with an inhibitor of ATP-adenosine A2aR- or ATP-adenosine A2bR-mediated signaling (such as an A2aR antagonist, an A2bR antagonist, and / or a CD73 inhibitor and / or a CD38 inhibitor and / or a CD39 inhibitor) and / or a Treg-attenuating agent. In some embodiments, the GPR174 inhibitor inhibits GPR174-mediated signaling via the Gsα subunit of heterotrimeric G proteins, which stimulates the 3′,5′-cyclic adenosine monophosphate (cAMP)-dependent pathway. The cAMP-dependent pathway (also referred to as Gs signaling or the Gs pathway) involves the synthesis of the second messenger cAMP, which then activates cAMP-dependent protein kinase (protein kinase A or PKA).

[0019] In one aspect, the present disclosure provides a pharmaceutical composition comprising an inhibitor of GPR174 signaling in combination with at least one or more of an adenosine A2A receptor antagonist, an adenosine A2B receptor antagonist, a CD73 inhibitor, and / or a CD38 inhibitor and / or a CD39 inhibitor, and a pharmaceutically acceptable carrier.

[0020] In one aspect, the present disclosure provides a pharmaceutical composition comprising a combination of an inhibitor of GPR174 signaling and a Treg attenuating agent, and a pharmaceutically acceptable carrier.

[0021] In some embodiments, the GPR174 inhibitor is a small molecule inhibitor that is an inverse agonist of GPR174 signaling.

[0022] In some embodiments, the small molecule inhibitor is an antagonist of GPR174 signaling.

[0023] In some embodiments, the small molecule inhibitor inhibits phosphatidylserine (PS)-dependent or lysophosphatidylserine (LysoPS)-dependent or pepducin-dependent activation of GPR174 signaling by at least 25% in cells expressing GPR174.

[0024] In some embodiments, the stimulated immune response comprises a T cell-mediated immune response.

[0025] In some embodiments, the T cell-mediated immune response comprises suppression of the activity, differentiation, growth, and / or proliferation of T-Regs. In some embodiments, the T cell-mediated immune response comprises stimulation of the activity, differentiation, growth, and / or proliferation of T effectors. In some embodiments, the T cell-mediated immune response comprises stimulation of the activity, differentiation, growth, and / or proliferation of Th1 cells. In some embodiments, the T cell-mediated immune response comprises suppression of the activity, differentiation, growth, and / or proliferation of Th17 cells. In one embodiment, the T cell-mediated immune response comprises preferential proliferation and activation of T effector cells over proliferation and activation of T regulatory cells, resulting in a relative decrease in the frequency, abundance, and / or function of T regulatory cells.

[0026] In some embodiments, the stimulated immune response comprises a decrease in immune cell-associated or cancer cell-associated programmed death-ligand 1 (PD-L1) expression, or cytotoxic T-lymphocyte-associated antigen 4 (CTLA4) expression, or T-cell immunoreceptor with Ig and ITIM domains (TIGIT) expression, or amphiregulin (AREG) expression.

[0027] In some embodiments, the stimulated immune response comprises an NK cell-mediated immune response. In some embodiments, at least a portion of the T cells or NK cells express GPR174. In some embodiments, the NK cell-mediated immune response comprises stimulation of NK cell function, activity, differentiation, growth, and / or proliferation.

[0028] In some embodiments, the stimulated and / or amplified immune response comprises increased Th1 cytokine production in the presence of cancer cell-derived exosomes.

[0029] In some embodiments, the patient suffers from or has a neoplasm that is not recognized as self by the body. In some embodiments, the neoplasm is a non-malignant neoplasm, such as a non-malignant tumor. In some embodiments, the neoplasm is a malignant tumor. In some embodiments, the neoplasm (either malignant or non-malignant) contains T cell infiltrates. In some embodiments, the tumor cells are destroyed by an immune response stimulated by a GPR174 inhibitory compound as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent.

[0030] In some embodiments, the patient is a cancer patient. In some embodiments, the mammalian patient is a human. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a hematological cancer. In some embodiments, the cancer is as disclosed herein. In some embodiments, at least some of the cancer cells express GPR174. In some embodiments, GPR174 has 90% or more than 90% sequence identity to SEQ ID NO:1.

[0031] In some embodiments, T cell activity, differentiation, proliferation, and / or growth is stimulated in a population of peripheral blood mononuclear cells (PBMCs) contacted with a GPR174 small molecule inhibitor as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to a control population of PBMCs not contacted with the GPR174 small molecule inhibitor.

[0032] In some embodiments, FoxP3 + The percentage of Helios+ T cells (regulatory T cells) is reduced by at least 20% in a population of PBMCs contacted with a GPR174 small molecule inhibitor as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to a control population of PBMCs not contacted with the small molecule GPR174 inhibitor.

[0033] In some embodiments, cAMP levels are reduced by at least 20%, at least 50%, or at least 75% in cells expressing GPR174 contacted with a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to control cells expressing GPR174 that have not been contacted with the GPR174 small molecule inhibitor.

[0034] In some embodiments, the activity of protein kinase A (PKA) is reduced by at least 20% in cells expressing GPR174 contacted with a small molecule inhibitor of GPR174 as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to control cells not contacted with the GPR174 small molecule inhibitor.

[0035] In some embodiments, PKA activity is reduced by at least 20%, at least 50%, or at least 75% in cells expressing GPR174 contacted with a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to control cells expressing GPR174 that have not been contacted with the GPR174 small molecule inhibitor.

[0036] In some embodiments, production of one or more of IL-2, IFN-γ, TNF-α, IL-6, and IL-10 is increased by at least 20% in peripheral blood mononuclear cells (PBMCs) contacted with a small molecule inhibitor of GPR174 compared to control cells not contacted with the small molecule inhibitor.

[0037] In some embodiments, production of one or more of IL-2, IFN-γ, TNF-α, and GM-CSF is increased by at least 20% in peripheral blood mononuclear cells (PBMCs) contacted with a small molecule inhibitor of GPR174 in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to control cells not contacted with the small molecule inhibitor of GPR174.

[0038] In some embodiments, the production of IL-17A is reduced by at least 20% in PBMCs contacted with a small molecule inhibitor compared to control cells not contacted with the small molecule inhibitor.

[0039] In some embodiments, production of one or more cytokines is reduced by at least 20%, at least 50%, or at least 80% in a population of PBMCs exposed to a small molecule inhibitor of GPR174 signaling compared to a population of PBMCs not exposed to the small molecule inhibitor of GPR174 signaling, hi some embodiments, production of IL-17A is reduced by at least 20%, at least 50%, or at least 80% in a population of PBMCs exposed to a small molecule inhibitor of GPR174 signaling compared to a population of PBMCs not exposed to the small molecule inhibitor of GPR174 signaling.

[0040] In some embodiments, production of one or more cytokines is increased by at least 20%, at least 50%, or at least 80% in a population of PBMCs exposed to a small molecule inhibitor of GPR174 signaling compared to a population of PBMCs not exposed to the small molecule inhibitor of GPR174 signaling, hi some embodiments, the cytokine is selected from IL-2, IFN-γ, TNF, IL-6, and IL-10.

[0041] In some embodiments, production of one or more cytokines is increased by at least 20%, at least 50%, or at least 80% in a population of PBMCs exposed to a small molecule inhibitor of GPR174 in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to control cells not contacted with the small molecule inhibitor of GPR174. In some embodiments, the cytokine is a T helper type 1 cell (Th1) cytokine selected from IL-2, IFN-γ, TNF, and GM-CSF.

[0042] In some embodiments, production of one or more cytokines is increased by at least 20%, at least 50%, at least 100%, at least 500%, or at least 1000% in a population of PBMCs exposed to a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to a population of PBMCs not exposed to the small molecule inhibitor of GPR174 signaling. In some embodiments, the cytokine is selected from IL-2, IFN-γ, TNFR, F, and GM-CSF. In some embodiments, production of one or more of IL-2, IFN-γ, TNF, or GM-CSF is increased by at least 20%, at least 50%, at least 100%, at least 500%, or at least 1000% in a population of PBMCs exposed to a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to a population of PBMCs not exposed to the small molecule inhibitor of GPR174 signaling.

[0043] In some embodiments, production of IL-2 is increased by at least 100%, at least 500%, or at least 1000% in a population of PBMCs exposed to a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to a population of PBMCs not exposed to the small molecule inhibitor of GPR174 signaling. In some embodiments, IFN-γ production is increased by at least 50%, at least 100%, or at least 500% in a population of PBMCs exposed to a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to a population of PBMCs not exposed to the small molecule inhibitor of GPR174 signaling. In some embodiments, TNF-α production is increased by at least 50%, at least 100%, or at least 500% in a population of PBMCs exposed to a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to a population of PBMCs not exposed to the small molecule inhibitor of GPR174 signaling.

[0044] In some embodiments, production of GM-CSF is increased by at least 50%, at least 100%, or at least 500% in a population of PBMCs exposed to a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to a population of PBMCs not exposed to the small molecule inhibitor of GPR174 signaling.

[0045] In some embodiments, production of one or more cytokines is unregulated and is not increased or decreased by more than 20% in a population of PBMCs exposed to a small molecule inhibitor of GPR174 signaling compared to a population of PBMCs not exposed to the small molecule inhibitor of GPR174 signaling, hi some embodiments, the one or more cytokines are selected from the group consisting of IL-4, IL-5, IL-9, IL-13, IL-17F, IL-21, and IL-22.

[0046] In some embodiments, the production of one or more cytokines is reduced by at least 20%, at least 50%, or at least 80% in cells expressing GPR174 contacted with a small molecule inhibitor compared to control cells expressing GPR174 that have not been contacted with a small molecule inhibitor, hi some embodiments, the production of IL-17A is reduced by at least 20%, at least 50%, or at least 80% in cells expressing GPR174 contacted with a small molecule inhibitor compared to control cells expressing GPR174 that have not been contacted with a small molecule inhibitor.

[0047] In some embodiments, production of one or more cytokines is increased by at least 20%, at least 50%, at least 100%, at least 500%, or at least 1000% in cells expressing GPR174 contacted with a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to control cells expressing GPR174 that have not been contacted with the small molecule inhibitor of GPR174. In some embodiments, production of one or more of IL-2, IFN-γ, TNF, or GM-CSF is increased by at least 20%, at least 50%, at least 100%, at least 500%, or at least 1000% in cells expressing GPR174 that have been contacted with a small molecule inhibitor of GPR174 compared to control cells expressing GPR174 that have not been contacted with the small molecule inhibitor of GPR174. In some embodiments, IL-2 production is increased by at least 100%, at least 500%, or at least 1000% in cells expressing GPR174 contacted with a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to control cells expressing GPR174 that have not been contacted with the small molecule inhibitor of GPR174. In some embodiments, IFN-γ production is increased by at least 50%, at least 100%, or at least 500% in cells expressing GPR174 contacted with a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to control cells expressing GPR174 that have not been contacted with the small molecule inhibitor of GPR174.In some embodiments, TNF-α production is increased by at least 50%, at least 100%, or at least 500% in cells expressing GPR174 contacted with a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to control cells expressing GPR174 that have not been contacted with the small molecule inhibitor of GPR174.

[0048] In some embodiments, production of one or more of IL-4, IL-5, IL-9, IL-13, IL-17F, IL-21, and IL-22 is unregulated and is not increased or decreased by more than 20% in cells expressing GPR174 contacted with a small molecule inhibitor of GPR174 signaling compared to control cells expressing GPR174 that have not been contacted with the small molecule inhibitor. In some embodiments, the patient has one or more tumors that may or may not contain infiltrating regulatory T cells.

[0049] In some aspects, provided herein are methods for inhibiting cAMP levels in cells expressing GPR174, comprising contacting the cells with a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent.

[0050] In some embodiments, cAMP levels are reduced by at least 20%, at least 50%, or at least 75% in cells expressing GPR174 contacted with a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to control cells expressing GPR174 that have not been contacted with the small molecule inhibitor of GPR174.

[0051] In some embodiments, the contacting is in vivo. In some embodiments, the contacting is in vitro. In some embodiments, the contacting is ex vivo.

[0052] In some embodiments, the cell is an immune cell. In some embodiments, the cell is a T cell or an NK cell. In some embodiments, the activity, differentiation, growth, and / or proliferation of an NK cell or a T cell is stimulated. In some embodiments, the cell is a mammalian cell. In some embodiments, the cell is a human cell. In some embodiments, the cell is a thymocyte, a lymph node cell, a spleen cell, a bone marrow cell, or a PMBC cell.

[0053] In some embodiments, GPR174 has 90% or greater than 90%, such as 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, to SEQ ID NO: 1. In some embodiments, GPR174 has the sequence set forth as SEQ ID NO: 1.

[0054] In some aspects, provided herein are methods for modulating the levels of one or more cytokines in PBMCs, comprising contacting the PBMCs with a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuator.

[0055] In some embodiments, the contacting is in vivo. In some embodiments, the contacting is in vitro. In some embodiments, the contacting is ex vivo.

[0056] In some embodiments, the PBMCs comprise immune cells. In some embodiments, the immune cells comprise T cells or NK cells. In some embodiments, the activity, differentiation, growth, and / or proliferation of NK cells or T cells is stimulated. In some embodiments, the PBMCs comprise mammalian cells. In some embodiments, the PBMCs comprise human cells.

[0057] In some embodiments, GPR174 has 90% or greater than 90% sequence identity to SEQ ID NO:1.

[0058] In some embodiments, the cytokine is selected from IL-2, IFN-γ, TNF, GM-CSF, IL-6, IL-17A, IL-10, IL-4, IL-5, IL-9, IL-12, IL-13, IL-17F, IL-21, and IL-22. In some embodiments, the level of one or more cytokines is increased. In some embodiments, the level of one or more cytokines selected from the group consisting of IL-2, IFN-γ, TNF, and GM-CSF is increased. In some embodiments, the level of one or more cytokines is decreased. In some embodiments, the level of one or more cytokines is not modulated.

[0059] In some embodiments, the level of one or more cytokines is reduced by at least 20%, at least 50%, or at least 80% in a population of PBMCs exposed to a small molecule inhibitor of GPR174 signaling compared to a population of PBMCs not exposed to the small molecule inhibitor of GPR174 signaling, hi some embodiments, the level of IL-17A is reduced by at least 20%, at least 50%, or at least 80% in a population of PBMCs exposed to a small molecule inhibitor of GPR174 signaling compared to a population of PBMCs not exposed to the small molecule inhibitor of GPR174 signaling.

[0060] In some embodiments, the level of one or more cytokines is increased by at least 20%, at least 50%, or at least 80% in a population of PBMCs exposed to a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to a population of PBMCs not exposed to the small molecule inhibitor of GPR174 signaling. In some embodiments, the cytokine is selected from IL-2, IFN-γ, TNF, and GM-CSF.

[0061] In some embodiments, the level of one or more cytokines is increased by at least 20%, at least 50%, at least 100%, at least 500%, or at least 1000% in a population of PBMCs exposed to a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to a population of PBMCs not exposed to the small molecule inhibitor of GPR174 signaling. In some embodiments, the cytokine is selected from IL-2, IFN-γ, TNF, and GM-CSF. In some embodiments, levels of one or more of IL-2, IFN-γ, TNF-α, or GM-CSF are increased by at least 20%, at least 50%, at least 100%, at least 500%, or at least 1000% in a population of PBMCs exposed to a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to a population of PBMCs not exposed to the small molecule inhibitor of GPR174 signaling.

[0062] In some embodiments, the level of IL-2 is increased by at least 100%, at least 500%, or at least 1000% in a population of PBMCs exposed to a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to a population of PBMCs not exposed to the small molecule inhibitor of GPR174 signaling. In some embodiments, the level of IFN-γ is increased by at least 50%, at least 100%, or at least 500% in a population of PBMCs exposed to a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to a population of PBMCs not exposed to the small molecule inhibitor of GPR174 signaling. In some embodiments, the level of TNF-α is increased by at least 50%, at least 100%, or at least 500% in a population of PBMCs exposed to a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to a population of PBMCs not exposed to the small molecule inhibitor of GPR174 signaling.

[0063] In some embodiments, the level of one or more cytokines is not modulated and is not increased or decreased by more than 20% in a population of PBMCs exposed to a small molecule inhibitor of GPR174 signaling compared to a population of PBMCs not exposed to the small molecule inhibitor of GPR174 signaling. In some embodiments, the one or more cytokines are selected from the group consisting of IL-4, IL-5, IL-9, IL-13, IL-17F, IL-21, and IL-22.

[0064] In some embodiments, the level of one or more cytokines is reduced by at least 20%, at least 50%, or at least 80% in cells expressing GPR174 contacted with a small molecule inhibitor compared to control cells expressing GPR174 that have not been contacted with a small molecule inhibitor, hi some embodiments, the level of IL-17A is reduced by at least 20%, at least 50%, or at least 80% in cells expressing GPR174 contacted with a small molecule inhibitor compared to control cells expressing GPR174 that have not been contacted with a small molecule inhibitor.

[0065] In some embodiments, the level of one or more cytokines is increased by at least 20%, at least 50%, at least 100%, at least 500%, or at least 1000% in cells expressing GPR174 contacted with a small molecule inhibitor of GPR174 as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to control cells expressing GPR174 that have not been contacted with the small molecule inhibitor of GPR174. In some embodiments, the levels of one or more of IL-2, IFN-γ, TNF, or GM-CSF are increased by at least 20%, at least 50%, at least 100%, at least 500%, or at least 1000% in cells expressing GPR174 contacted with a small molecule inhibitor of GPR174 as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to control cells expressing GPR174 that have not been contacted with the small molecule inhibitor of GPR174.

[0066] In some embodiments, the level of IL-2 is increased by at least 100%, at least 500%, or at least 1000% in cells expressing GPR174 contacted with a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to control cells expressing GPR174 that have not been contacted with the small molecule inhibitor of GPR174.

[0067] In some embodiments, the level of IFN-γ is increased by at least 50%, at least 100%, or at least 500% in cells expressing GPR174 contacted with a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to control cells expressing GPR174 that have not been contacted with a small molecule inhibitor of GPR174.

[0068] In some embodiments, the level of TNF (also referred to as "TNF-α") is increased by at least 50%, at least 100%, or at least 500% in cells expressing GPR174 that have been contacted with a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to control cells expressing GPR174 that have not been contacted with the small molecule inhibitor of GPR174.

[0069] In some embodiments, the levels of one or more of IL-4, IL-5, IL-9, IL-13, IL-17F, IL-21, and IL-22 are not regulated and do not increase or decrease by more than 20% in cells expressing GPR174 that have been contacted with a small molecule inhibitor of GPR174 signaling compared to control cells expressing GPR174 that have not been contacted with the small molecule inhibitor.

[0070] In some aspects, provided herein is a pharmaceutical composition comprising a small molecule inhibitor of GPR174 signaling and a pharmaceutically acceptable excipient.

[0071] In some aspects, provided herein are pharmaceutical compositions comprising a small molecule inhibitor of GPR174 signaling in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, and / or a CD73 inhibitor, and a pharmaceutically acceptable excipient.

[0072] In some aspects, provided herein is a pharmaceutical composition comprising a small molecule inhibitor of GPR174 signaling in combination with an A2aR antagonist, and a pharmaceutically acceptable excipient.

[0073] In some aspects, provided herein is a pharmaceutical composition comprising a small molecule inhibitor of GPR174 signaling in combination with an A2bR antagonist, and a pharmaceutically acceptable excipient.

[0074] In some aspects, provided herein are pharmaceutical compositions comprising a small molecule inhibitor of GPR174 signaling in combination with an A2aR antagonist and an A2bR antagonist, and a pharmaceutically acceptable excipient.

[0075] In some aspects, provided herein is a pharmaceutical composition comprising a small molecule inhibitor of GPR174 signaling in combination with a Treg attenuating agent, and a pharmaceutically acceptable excipient.

[0076] In some embodiments, provided herein are pharmaceutical compositions comprising a small molecule inhibitor of GPR174, (i) at least one of an A2aR antagonist and / or an A2bR antagonist, and (ii) at least one of a CD73 inhibitor and / or a CD38 inhibitor and / or a CD39 inhibitor. In some embodiments, the pharmaceutical composition further comprises a Treg attenuating agent.

[0077] In some embodiments, the small molecule inhibitor of GPR174 does not include an alkyl chain containing 10 or more C atoms. In some embodiments, the small molecule inhibitor of GPR174 is not a pepducin, LysoPS, PS, or a compound disclosed in U.S. Patent Application Publication No. 2015 / 0361119. In some embodiments, the small molecule inhibitor has a molecular weight of about 50 Da to about 2500 Da. In some embodiments, the small molecule inhibitor has a molecular weight of about 50 to 800 Da, e.g., about 100 Da to about 800 Da.

[0078] In some embodiments, the small molecule inhibitor alters the cellular distribution of a recombinant GPR174 polypeptide modified to include a nuclear localization signal compared to the cellular distribution of the recombinant GPR174 polypeptide in the absence of the inhibitor. In some embodiments, the small molecule inhibitor competitively binds to GPR174 compared to any one of compounds 1-59. In some embodiments, the small molecule inhibitor has an EC of 15 μM for GPR174 as measured by a cellular redistribution assay (CRA) of less than 15 μM. 50CRAs are described in detail herein under "Screening Methods." In some embodiments, the small molecule inhibitor competitively binds to GPR174 compared to any one of compounds 1-59, resulting in a change in the apparent binding affinity of the compound to GPR174 compared to the binding affinity of the compound to GPR174 alone when cells expressing GPR174 are simultaneously contacted with the compound and the small molecule inhibitor. In some embodiments, the small molecule inhibitor causes a difference in the inhibitory activity of any one of reference compounds 1-59 in a GPR174-mediated signaling assay compared to the inhibitory activity of the reference compound alone when tested in the presence of said reference compound. In some embodiments, the small molecule inhibitor binds allosterically to GPR174 compared to any one of compounds 1-59. In some embodiments, the small molecule inhibitor is specific for GPR174 compared to one or more GPCRs in the reference panel of GPCRs disclosed herein. In some embodiments, the small molecule inhibitor is specific for GPR174 compared to P2Y10. In some embodiments, the small molecule inhibitor is specific for GPR174 compared to GPR34. In some embodiments, the small molecule inhibitor inhibits the Gs signaling pathway.

[0079] In some embodiments, T cell activity, differentiation, proliferation, and / or growth is stimulated in a population of peripheral blood mononuclear cells (PBMCs) contacted with a small molecule inhibitor of GPR174 as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to a control population of PBMCs not contacted with the small molecule inhibitor of GPR174.

[0080] In some embodiments, a subpopulation of regulatory T cells is decreased in a population of PBMCs contacted with a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuator, compared to a control population of PBMCs not contacted with a small molecule inhibitor of GPR174.

[0081] In some embodiments, FoxP3 + The percentage of Helios+ T cells (regulatory T cells) is reduced by at least 20% in a population of PBMCs contacted with a small molecule inhibitor of GPR174 as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to a control population of PBMCs not contacted with a small molecule inhibitor of GPR174.

[0082] In some embodiments, cAMP levels are reduced by at least 20% in cells expressing GPR174 contacted with a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to control cells expressing GPR174 that have not been contacted with the small molecule inhibitor of GPR174.

[0083] In some embodiments, cAMP levels are reduced by at least 20%, at least 50%, or at least 75% in cells expressing GPR174 contacted with a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to control cells expressing GPR174 that have not been contacted with the small molecule inhibitor of GPR174.

[0084] In some embodiments, PKA activity is reduced by at least 20% in cells expressing GPR174 contacted with a small molecule inhibitor of GPR174 as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to control cells not contacted with the small molecule inhibitor of GPR174.

[0085] In some embodiments, PKA activity is reduced by at least 20%, at least 50%, or at least 75% in cells expressing GPR174 contacted with a small molecule inhibitor of GPR174 as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to control cells expressing GPR174 that have not been contacted with the small molecule inhibitor of GPR174.

[0086] In some embodiments, the levels of one or more cytokines are modulated in PBMCs contacted with a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to control cells not contacted with the small molecule inhibitor of GPR174.

[0087] In some embodiments, levels of one or more of IL-2, IFN-γ, TNF, and GM-CSF are increased by at least 20% in PBMCs contacted with a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuator, compared to control cells not contacted with the small molecule inhibitor of GPR174.

[0088] In some embodiments, the level of IL-17A is reduced by at least 20% in PBMCs contacted with a small molecule inhibitor compared to control cells not contacted with the small molecule inhibitor.

[0089] In some embodiments, the level of one or more cytokines is reduced by at least 20%, at least 50%, or at least 80% in a population of PBMCs exposed to a small molecule inhibitor of GPR174 signaling compared to a population of PBMCs not exposed to the small molecule inhibitor of GPR174 signaling, hi some embodiments, the level of IL-17A is reduced by at least 20%, at least 50%, or at least 80% in a population of PBMCs exposed to a small molecule inhibitor of GPR174 signaling compared to a population of PBMCs not exposed to the small molecule inhibitor of GPR174 signaling.

[0090] In some embodiments, the level of one or more cytokines is increased by at least 20%, at least 50%, or at least 80% in a population of PBMCs exposed to a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to a population of PBMCs not exposed to the small molecule inhibitor of GPR174 signaling. In some embodiments, the cytokine is selected from IL-2, IFN-γ, TNF, and GM-CSF.

[0091] In some embodiments, the level of one or more cytokines is increased by at least 20%, at least 50%, at least 100%, at least 500%, or at least 1000% in a population of PBMCs exposed to a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to a population of PBMCs not exposed to the small molecule inhibitor of GPR174 signaling. In some embodiments, the cytokine is selected from IL-2, IFN-γ, TNF, and GM-CSF. In some embodiments, the levels of one or more of IL-2, IFN-γ, TNF, or GM-CSF are increased by at least 20%, at least 50%, at least 100%, at least 500%, or at least 1000% in a population of PBMCs exposed to a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to a population of PBMCs not exposed to the small molecule inhibitor of GPR174 signaling.

[0092] In some embodiments, the level of IL-2 is increased by at least 100%, at least 500%, or at least 1000% in a population of PBMCs exposed to a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to a population of PBMCs not exposed to the small molecule inhibitor of GPR174 signaling. In some embodiments, the level of IFN-γ is increased by at least 50%, at least 100%, or at least 500% in a population of PBMCs exposed to a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to a population of PBMCs not exposed to the small molecule inhibitor of GPR174 signaling. In some embodiments, the level of TNF-α is increased by at least 50%, at least 100%, or at least 500% in a population of PBMCs exposed to a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to a population of PBMCs not exposed to the small molecule inhibitor of GPR174 signaling.

[0093] In some embodiments, the level of one or more cytokines is not modulated and is not increased or decreased by more than 20% in a population of PBMCs exposed to a small molecule inhibitor of GPR174 signaling compared to a population of PBMCs not exposed to the small molecule inhibitor of GPR174 signaling. In some embodiments, the one or more cytokines are selected from the group consisting of IL-4, IL-5, IL-9, IL-13, IL-17F, IL-21, and IL-22.

[0094] In some embodiments, the level of one or more cytokines is reduced by at least 20%, at least 50%, or at least 80% in immune cells contacted with a small molecule inhibitor compared to control cells expressing GPR174 that have not been contacted with the small molecule inhibitor, hi some embodiments, the level of IL-17A is reduced by at least 20%, at least 50%, or at least 80% in immune cells contacted with a small molecule inhibitor compared to control cells that have not been contacted with the small molecule inhibitor.

[0095] In some embodiments, the level of one or more cytokines is increased by at least 20%, at least 50%, at least 100%, at least 500%, or at least 1000% in immune cells contacted with a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to control cells not contacted with the small molecule inhibitor of GPR174. In some embodiments, levels of one or more of IL-2, IFN-γ, TNF, or GM-CSF are increased by at least 20%, at least 50%, at least 100%, at least 500%, or at least 1000% in immune cells contacted with a small molecule inhibitor of GPR174 as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to control cells not contacted with the small molecule inhibitor of GPR174.

[0096] In some embodiments, the level of IL-2 is increased by at least 100%, at least 500%, or at least 1000% in immune cells contacted with a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to control cells not contacted with the small molecule inhibitor of GPR174. In some embodiments, the level of IFN-γ is increased by at least 50%, at least 100%, or at least 500% in immune cells contacted with a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to control cells not contacted with the small molecule inhibitor of GPR174. In some embodiments, the level of TNF-α is increased by at least 50%, at least 100%, or at least 500% in immune cells contacted with a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to control cells not contacted with the small molecule inhibitor of GPR174.

[0097] In some embodiments, the levels of one or more of IL-4, IL-5, IL-9, IL-13, IL-17F, IL-21, and IL-22 are not regulated and are not increased or decreased by more than 20% in immune cells contacted with a small molecule inhibitor of GPR174 signaling compared to control cells not contacted with the small molecule inhibitor.

[0098] In some embodiments, the small molecule inhibitor reduces ligand-induced receptor internalization of GPR174 in cells expressing GPR174 compared to control cells not contacted with the small molecule inhibitor.

[0099] In some embodiments, receptor internalization is reduced by at least 10%, at least 20%, at least 50%, or at least 80% in cells expressing GPR174 contacted with a small molecule inhibitor of GPR174 signaling compared to control cells expressing GPR174 that have not been contacted with the small molecule inhibitor. In some embodiments, receptor internalization is reduced by 1-99% in cells expressing GPR174 contacted with a small molecule inhibitor of GPR174 signaling compared to control cells expressing GPR174 that have not been contacted with the small molecule inhibitor. In some embodiments, receptor internalization is reduced by 1-20%, 20-40%, 40-60%, 60-80%, or 80-99% in cells expressing GPR174 contacted with a small molecule inhibitor of GPR174 signaling compared to control cells expressing GPR174 that have not been contacted with the small molecule inhibitor.

[0100] In some embodiments, β-arrestin recruitment is reduced by at least 10%, at least 20%, at least 50%, or at least 80% in cells expressing GPR174 contacted with a small molecule inhibitor of GPR174 signaling compared to control cells expressing GPR174 that have not been contacted with the small molecule inhibitor. In some embodiments, β-arrestin recruitment is reduced by 1-99% in cells expressing GPR174 contacted with a small molecule inhibitor of GPR174 signaling compared to control cells expressing GPR174 that have not been contacted with the small molecule inhibitor. In some embodiments, β-arrestin recruitment is reduced by 1-20%, 20-40%, 40-60%, 60-80%, or 80-99% in cells expressing GPR174 contacted with a small molecule inhibitor of GPR174 signaling compared to control cells expressing GPR174 that have not been contacted with the small molecule inhibitor.

[0101] In some aspects, provided herein are compositions comprising a small molecule inhibitor of GPR174, either as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, and a cell expressing GPR174.

[0102] In some embodiments, the cell is a mammalian cell. In some embodiments, the cell is a human cell. In some embodiments, the cell is an immune cell. In some embodiments, the cell is a T cell. In some embodiments, the T cell is a naive T cell. In some embodiments, the T cell is a regulatory T cell. In some embodiments, the T cell is an effector T cell. In some embodiments, the cell is a thymocyte, lymph node cell, spleen cell, bone marrow cell, or PBMC cell. In some embodiments, the level of cAMP is reduced by at least 20% in the composition compared to control cells expressing GPR174 that have not been contacted with a small molecule inhibitor of GPR174. In some embodiments, the activity of PKA is reduced by at least 20% in the composition compared to control cells expressing GPR174 that have not been contacted with a small molecule inhibitor of GPR174. In some embodiments, the level of one or more cytokines is modulated in the composition compared to control cells expressing GPR174 that have not been contacted with a small molecule inhibitor of GPR174.

[0103] In some embodiments, cAMP levels are reduced by at least 20%, at least 50%, or at least 75% in cells expressing GPR174 contacted with a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to control cells expressing GPR174 that have not been contacted with the small molecule inhibitor of GPR174.

[0104] In some embodiments, PKA activity is reduced by at least 20%, at least 50%, or at least 75% in cells expressing GPR174 contacted with a small molecule inhibitor of GPR174 signaling as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, compared to control cells expressing GPR174 that have not been contacted with the small molecule inhibitor of GPR174.

[0105] In some aspects, provided herein are compositions comprising a small molecule inhibitor of GPR174 signaling, either as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, and a population of cells, at least a portion of which express GPR174 and at least a portion of which expresses GPR174 comprises T cells.

[0106] In some embodiments, the population of cells comprises cancer cells. In some embodiments, the population of T cells comprises regulatory T cells. In some embodiments, the population of T cells comprises effector T cells.

[0107] In another aspect, the disclosure features a method for inhibiting a GPR174-mediated signaling pathway (e.g., a Gs signaling pathway) in a cell. The method includes contacting a cell with an isolated compound previously determined to functionally interact with GPR174, either as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, wherein the cell comprises a GPR174-mediated signaling pathway and expresses GPR174 (e.g., a GPR174 polypeptide), and the isolated compound inhibits the ability of GPR174 to regulate the GPR174-mediated signaling pathway, thereby inhibiting the GPR174-mediated signaling pathway in the cell. In certain embodiments, the compound can functionally interact with human GPR174 having the sequence set forth as SEQ ID NO:1, or with a naturally occurring variant of GPR174 having at least 90% identity to SEQ ID NO:1. In certain embodiments, the isolated compound is capable of altering the cellular distribution of a recombinant GPR174 polypeptide (e.g., human GPR174 having the sequence of SEQ ID NO: 1) that has been modified to alter the cellular distribution compared to the cellular distribution of the recombinant GPR174 polypeptide in the absence of the compound. In some embodiments, the cells are contacted in vivo in a subject suffering from a disease such as cancer.

[0108] In a related aspect, the disclosure features a method for inhibiting a GPR174-mediated signaling pathway in a cell by (i) providing an isolated compound that has been predetermined to functionally interact with human GPR174 and inhibit a GPR174-mediated signaling pathway in a cell that expresses GPR174, and (ii) contacting a cell that expresses GPR174 with the isolated compound as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, thereby inhibiting the GPR174-mediated signaling pathway in the cell. In some embodiments, the cells express human GPR174 having the sequence set forth as SEQ ID NO:1, or a naturally occurring variant of GPR174 having at least 90% identity to SEQ ID NO:1 (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:1), and inhibit GPR174-mediated signaling pathways in cells expressing GPR174. In some embodiments, the method comprises contacting a cell expressing human GPR174 with an isolated compound previously determined to functionally interact with human GPR174 having the sequence set forth as SEQ ID NO: 1 or a naturally occurring variant of GPR174 having at least 90% (such as at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) identity to SEQ ID NO: 1, and to inhibit a GPR174-mediated signaling pathway in the cell expressing GPR174. In some embodiments, step (i) comprises performing at least one assay to measure the functional interaction of the compound with GPR174 prior to contacting the cell by step (ii).

[0109] In either aspect, the compound optionally competitively binds the same region of GPR174 as any one of compounds 1-59 provided below.

[0110] In another aspect, the disclosure provides a method of using a small molecule chemical compound to inhibit a GPR174-mediated signaling pathway in a cell, comprising: (a) providing a small molecule chemical compound, wherein the small molecule chemical compound functionally interacts with GPR174 and inhibits the GPR174-mediated Gs signaling pathway in cells expressing GPR174, and the compound meets the following criteria: (i) the compound has a structure selected from the group consisting of Formulas I, II, III, IV, V, and VI (or (I), (II), (III), (IV), (V), (Va), or (VI)); or (ii) the compound alters the binding affinity of any one of reference compounds 1-59 (listed in Table 1) to GPR174; or (iii) the compound, when tested in the presence of the reference compound, differentially inhibits the inhibitory activity of any one of reference compounds 1-59 (listed in Table 1) in a GPR174-mediated signaling pathway assay compared to the inhibitory activity of the reference compound alone: characterized by at least one of the steps of: (b) contacting a cell expressing GPR174, including a GPR174-mediated Gs signaling pathway, with the compound of step (a) as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, thereby inhibiting the GPR174-mediated signaling pathway in the cell. The present invention is characterized by a method comprising:

[0111] In another aspect, the disclosure provides a method of using a small molecule chemical compound to inhibit a GPR174-mediated signaling pathway in a cell, comprising: (a) providing a small molecule chemical compound, wherein the small molecule chemical compound functionally interacts with GPR174 and inhibits the GPR174-mediated Gs signaling pathway in cells expressing GPR174, and the compound meets the following criteria: (i) the compound has a structure selected from the group consisting of Formulas I, II, III, IV, V, and VI (or (I), (II), (III), (IV), (V), (Va), or (VI)); or (ii) the compound alters the apparent binding affinity of any one of reference compounds 1-59 (listed in Table 1) to GPR174; or (iii) the compound differentially inhibits the inhibitory activity of any one of Reference Compounds 1-59 (listed in Table 1) in a GPR174-mediated signaling pathway assay compared to the inhibitory activity of the reference compound alone when tested in the presence of said reference compound: characterized by at least one of the steps of: (b) contacting a cell expressing GPR174, including a GPR174-mediated Gs signaling pathway, with the compound of step (a) as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, thereby inhibiting the GPR174-mediated signaling pathway in the cell. The present invention is characterized by a method comprising:

[0112] In some embodiments, the compound alters the binding affinity of any one of the compounds described herein (e.g., any one of Reference Compounds 1-59 in Table 1). In certain embodiments, the compound decreases (i.e., inhibits) the binding affinity of any one of Reference Compounds 1-59 to GPR174. In certain embodiments, the compound competitively decreases (i.e., competitively inhibits) the binding affinity of any one of Reference Compounds 1-59 to GPR174. In alternative embodiments, the compound increases the binding affinity of any one of Reference Compounds 1-59 to GPR174. In one embodiment, the compound has a structure selected from the group consisting of Formulas I-VI. In one embodiment, the compound inhibits lysophosphatidylserine (LysoPS)-dependent activation of the GPR174 receptor in the cell. In one embodiment, the compound inhibits lysophosphatidylserine (LysoPS)-dependent activation of the GPR174 receptor in the cell.

[0113] In one embodiment, the compound inhibits phosphatidylserine (PS)-dependent activation of the GPR174 receptor in the cells. As described in Examples 16 and 17 herein, the inventors have discovered that phosphatidylserine (PS) is an agonist of GPR174-mediated Gs signaling. Example 17 provides experimental results demonstrating that apoptotic cells stimulate the GPR174 Gs signaling pathway in cells expressing GPR174. As further described in Example 16 and shown in Figures 47A-47F, PS-mediated GPR174 Gs signaling is inhibited by representative GPR174 inhibitory compounds 6, 10, 11, 20, 23, and 30, which belong to different chemical classes (i.e., groups I, II, and IV). This data demonstrates that compounds 6, 10, 11, 20, 23, and 30 act as GPR174 antagonists and inhibit PS liposome-mediated cAMP signaling. Thus, GPR174-mediated PS liposome signaling is inhibited by multiple GPR174-inhibitory small molecule compounds with diverse chemical structures.

[0114] As described herein, extracellular PS is highly enriched in the tumor microenvironment and is found not only on tumor cells but also on the surface of endothelial cells in blood vessels that permeate solid tumors. Furthermore, apoptotic neutrophils and activated platelets, both of which expose PS, are also recruited to solid tumors (see AK and Rao DA, Blood 120:4667-4668, 2012; Schlesinger, M., Journal of Hematology and Oncology (11) 125, 2018; Treffers LW et al., Immunological Reviews vol. 273:312-328, 2016; and Gregory AD and Houghton AM, Cancer Research vol. 71(7):2411-6, 2011). Therefore, high concentrations of PS are considered a major source of tumor-mediated immunosuppression and may play a role in resistance to cancer immunotherapies, such as checkpoint inhibitors.

[0115] Thus, the experimental results described in Examples 16 and 17 provide support for a method of stimulating T cell-mediated immunity in a subject suffering from cancer, comprising administering to the subject a GPR174 inhibitor, either as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg-attenuating agent, in an amount effective to stimulate T cell-mediated immunity as described herein.

[0116] As further described herein, the present inventors have demonstrated that cancer cell-derived exosomes stimulate GPR174, and that inhibition of GPR174 in the presence of cancer cell-derived exosomes increases Th1 cytokine levels to a greater extent than in the presence of PS liposomes, as described in Example 26. Thus, in one aspect, there is provided a method for stimulating and / or amplifying an immune response in a mammalian subject suffering from cancer or cancer metastasis, or at risk of developing cancer or cancer metastasis, comprising: (a) determining the presence of cancer cell-derived exosomes expressing phosphatidylserine (PS) in a sample obtained from the mammalian subject; and (b) administering to the subject a therapeutically effective amount of a GPR174 inhibitor. A method is provided, comprising:

[0117] In some embodiments, the GPR174 inhibitor is a small molecule that inhibits PS-mediated immunosuppression. In some embodiments, the sample is a tissue, cell, or cell extract, or a bodily fluid selected from the group consisting of blood, serum, plasma, sputum, urine, saliva, and tears. In some embodiments, step (a) comprises contacting the sample or an exosome extract obtained from the sample with a PS-binding substance to thereby determine the presence of cancer cell-derived exosomes. Because the exosome surface membrane reflects the plasma membrane of their parent cell, tumor cell-derived exosomes, in contrast to exosomes derived from normal cells, are characterized by having phosphatidylserine (PS) on their surface. Exemplary PS-binding substances include, for example, anti-PS antibodies or any other PS-binding substance, such as PKC, protein S, factor VII, factor III, and other known PS-binding substances. The PS-binding substance can be labeled with a detectable agent, such as a radioisotope, a colorimetric label, a fluorescent label, a magnetic resonance label, an enzyme, an affinity ligand, or a luminescent label. Exosomes can be extracted from a sample obtained from a subject by various means, such as selective precipitation, affinity purification, or differential centrifugation by known methods (see, for example, Zhang Y. et al., Mol Cell 39:133-144, 2010). In some embodiments, step (a) comprises determining a quantitative amount of cancer cell-derived exosomes in a sample obtained from the subject. In some embodiments, the method comprises determining whether the subject has an amount of cancer cell-derived exosomes above a threshold value indicating the presence of cancer (i.e., one or more tumors). Generally, normal tumor-free individuals have undetectable levels of PS-exosomes in unenriched samples. In contrast, patients with tumors typically exhibit values ​​of greater than 100 pg / 50 μL of sample in unenriched samples. Thus, a value greater than 50 pg / 50 μL plasma (or serum) indicates malignancy (i.e., cancer or one or more tumors).

[0118] It has been found that exosomes expressing PS are released from tumor cells, and circulating PS-positive tumor exosomes can be detected in biological fluids such as blood and urine.In some embodiments, the cancer cell-derived exosomes expressing PS are derived from lung cancer cells, pancreatic cancer cells, ovarian cancer cells, breast cancer cells, colon cancer cells, kidney cancer cells, liver cancer cells, skin cancer cells, brain cancer cells, head and neck cancer cells, or thyroid cancer cells, or any other cancer cell type disclosed herein.In some embodiments, the method further comprises administering to the subject at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent.

[0119] In some embodiments, the GPR174 inhibitory compound alters the apparent binding affinity of any one of the compounds described herein (e.g., any one of Reference Compounds 1-59 in Table 1) to GPR174. In certain embodiments, the compound decreases the apparent binding affinity of any one of Reference Compounds 1-59 to GPR174. In certain embodiments, the compound competitively binds to GPR174, decreasing the apparent binding affinity of any one of Reference Compounds 1-59 to GPR174. In alternative embodiments, the compound allosterically binds to GPR174, decreasing the apparent binding affinity of any one of Reference Compounds 1-59 to GPR174. In alternative embodiments, the compound increases the apparent binding affinity of any one of Reference Compounds 1-59 to GPR174. In one embodiment, the compound has a structure selected from the group consisting of Formulas I-VI. In one embodiment, the compound inhibits lysophosphatidylserine (LysoPS)-dependent activation of the GPR174 receptor in the cell. In one embodiment, the compound inhibits PS-dependent activation of the GPR174 receptor in the cell.

[0120] The cell expressing GPR174 can be a eukaryotic cell, such as a mammalian cell (e.g., a human cell). The cell can be present in a mammal (e.g., a human, a non-human primate, a rodent, a dog, a cat, a horse, a pig, or a cow). In some embodiments, the cell is contacted in vitro. In some embodiments, the cell is present in a mammal and is contacted in vivo. In some embodiments, the cell expressing GPR174 is an immune cell, such as a T cell. In certain embodiments, the mammal (e.g., a human) can suffer from cancer or be at risk of developing cancer.

[0121] In another aspect, the disclosure features a compound for use in treating cancer, characterized by interacting with GPR174 and inhibiting a GPR174-mediated signaling pathway (e.g., the Gs signaling pathway), wherein the GPR174 inhibitory compound is used to treat cancer as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent. In certain embodiments, the GPR174 inhibitory compound is also characterized by exhibiting the ability to alter the cellular distribution of a recombinant GPR174 polypeptide modified to include a nuclear localization signal, compared to the cellular distribution of the recombinant GPR174 polypeptide in the absence of the compound. In some embodiments, the method comprises contacting GPR174-expressing cancer cells and / or GPR174-expressing immune cells with an isolated compound, wherein the isolated compound has been previously determined to functionally interact with GPR174 and inhibit the GPR174-mediated signaling pathway, either as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuator. In some embodiments, the method comprises contacting GPR174-expressing immune cells, such as T cells, with a compound that inhibits the GPR174-mediated signaling pathway, either as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuator. In some embodiments, the methods include performing at least one assay to determine the functional interaction of said compound with GPR174 prior to administering said compound to the subject.In another aspect, the present disclosure features a compound for use in treating a tumor in a patient suffering from cancer, the compound interacting with GPR174 and inhibiting a GPR174-mediated signaling pathway (e.g., the Gs signaling pathway) as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent. In certain embodiments, the GPR174 inhibitory compound also exhibits the ability to alter the cellular distribution of a recombinant GPR174 polypeptide modified to include a nuclear localization signal, compared to the cellular distribution of the recombinant GPR174 polypeptide in the absence of the compound. In some embodiments, the tumor contains regulatory T cell (Treg) infiltration. In some embodiments, the tumor containing Treg infiltration is a solid organ tumor. In some embodiments, the tumor is present in a subject suffering from a cancer selected from the group consisting of breast cancer, lung cancer (such as small cell lung cancer or non-small cell lung cancer), colorectal cancer, cervical cancer, renal cancer, ovarian cancer, melanoma, pancreatic cancer, hepatocellular carcinoma, gastric cancer, glioblastoma, glioma, bladder cancer, myeloma (such as multiple myeloma), prostate cancer, thyroid cancer, testicular cancer, and esophageal cancer. In some embodiments, the method comprises performing at least one assay to measure the functional interaction of the compound with GPR174 before administering the compound to the subject.

[0122] In another aspect, the present disclosure features a method for treating or preventing cancer in a subject in need thereof. The method includes administering a compound to the subject, wherein the compound binds to and inhibits the GPR174 receptor, thereby inhibiting the GPR174-mediated signaling pathway, and the inhibition of the GPR174-mediated signaling pathway by the GPR174 inhibitor, optionally in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, results in the treatment or prevention of cancer. In some embodiments, the method includes administering an isolated compound to the subject, wherein the isolated compound is pre-determined to functionally interact with GPR174 and inhibit the GPR174-mediated signaling pathway, and the inhibition of the signaling pathway results in the treatment of cancer. In some embodiments, the GPR174 inhibitory compound meets the following criteria: (i) the compound has a structure selected from the group consisting of Formulas I-VI; (ii) the compound alters the binding affinity of any one of Reference Compounds 1-59 (listed in Table 1) to GPR174; and / or (iii) the compound differentially inhibits the inhibitory activity of any one of Reference Compounds 1-59 (listed in Table 1) in a GPR174-mediated signaling pathway assay compared to the inhibitory activity of the reference compound alone when tested in the presence of said reference compound: In some embodiments, the compound inhibits LysoPS-dependent activation of the GPR174 receptor in the subject. In some embodiments, the compound inhibits PS-dependent activation of the GPR174 receptor in the subject. In some embodiments, the GPR174 inhibitory compound satisfies the following criteria: (i) the compound has a structure selected from the group consisting of Formulas I-VI; (ii) the compound alters the apparent binding affinity of any one of the reference compounds 1 to 59 (listed in Table 1) to GPR174; and / or (iii) the compound, when tested in the presence of the reference compound, differentially inhibits the inhibitory activity of any one of reference compounds 1-59 (listed in Table 1) in a GPR174-mediated signaling pathway assay compared to the inhibitory activity of the reference compound alone: In some embodiments, the compound inhibits LysoPS-dependent or PS-dependent activation of the GPR174 receptor in the subject. In another aspect, the disclosure features a method of stimulating T cell-mediated immunity in a subject suffering from cancer, comprising administering to the subject a small molecule compound previously determined to inhibit the GPR174-mediated signaling pathway, either as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, and / or a CD73 inhibitor. In some embodiments, the compound inhibits GPR174-mediated signaling in regulatory T cells, thereby inhibiting the growth and / or proliferation and / or activity of regulatory T cells. In some embodiments, the compound inhibits GPR174-mediated signaling in effector T cells, thereby stimulating the growth and / or proliferation and / or cytotoxic activity of effector T cells. In some embodiments, the subject is afflicted with a tumor comprising regulatory T cells, and inhibition of GPR174 inhibits and / or suppresses the growth and / or proliferation of the regulatory T cells. In some embodiments of any of the above aspects, the cell is present in a mammal, and the mammal is afflicted with or at risk of developing cancer.

[0123] In some embodiments, the subject has or has a neoplasm that is not recognized as self by the body. In some embodiments, the neoplasm is a non-malignant neoplasm. In some embodiments, the neoplasm is a malignant neoplasm.

[0124] In some embodiments, the subject is diagnosed with an acoustic neuroma, anal cancer (including carcinoma in situ), squamous cell carcinoma, adrenal gland tumors (including adenoma, hyperaldosteronism, and adrenocortical carcinoma), Cushing's syndrome, benign paraganglioma, appendix cancer (including pseudomyxoma peritonei, carcinoid tumor, and non-carcinoid appendix tumor), bile duct cancer (including intrahepatic cholangiocarcinoma, extrahepatic cholangiocarcinoma, perihilar cholangiocarcinoma, and distal bile duct cancer), gallbladder cancer, bone cancer (including chondrosarcoma, osteosarcoma, malignant fibrous histiocytoma, fibrosarcoma, and chordoma), brain tumor (including craniopharyngioma, dermoid cyst, epidermoid tumor, Glioma, astrocytoma, low-grade astrocytoma, anaplastic astrocytoma, ependymoma, glioblastoma, oligodendroglioma, hemangioblastoma, pineal tumor, pituitary tumor, sarcoma, chordoma), breast cancer (including lobular carcinoma, triple-negative breast cancer, recurrent breast cancer, and brain metastasis), bladder cancer (including transitional cell bladder cancer, squamous cell carcinoma, and adenocarcinoma), cancer of unknown primary (CUP), (including adenocarcinoma, poorly differentiated carcinoma, squamous cell carcinoma, poorly differentiated malignant neoplasm, and neuroendocrine carcinoma), cervical cancer (including squamous cell carcinoma, adenocarcinoma, and mixed carcinoma) carcinoid tumors, childhood germ cell tumors (including yolk sac tumors, teratomas, embryonal carcinomas, polyembryoma, and germinomas), childhood brain tumors (including ependymoma, craniopharyngioma, chordoma, pleomorphic xanthoastrocytoma, meningioma, primitive neuroectodermal tumor, ganglioglioma, pineoblastoma, germ cell tumor, mixed glial-neuronal tumor, astrocytoma, and choroid plexus tumor), childhood leukemia (including lymphoblastic leukemia and myeloid leukemia), childhood blood disorders (Fanconi anemia, Diamond-Blackfan anemia, aplastic anemia, Shwachman-Diamond syndrome, Cox syndrome, and childhood liver cancer (including hepatoblastoma and hepatocellular carcinoma); childhood lymphoma (including Hodgkin lymphoma, non-Hodgkin lymphoma, Burkitt lymphoma, lymphoblastic lymphoma, and large cell lymphoma); childhood osteosarcoma; childhood melanoma; childhood soft tissue sarcoma; colorectal cancer (including adenocarcinoma, hereditary nonpolyposis colorectal cancer syndrome, and familial adenomatous polyposis); desmoplastic small round cell tumor (DSRCT);Esophageal cancer (including adenocarcinoma and squamous cell carcinoma), Ewing's sarcoma (including Ewing's sarcoma of bone, extraskeletal Ewing's tumor, and peripheral primitive neuroectodermal tumor), eye cancer (including uveal melanoma, basal cell carcinoma, squamous cell carcinoma, eyelid melanoma, conjunctival melanoma, sebaceous gland carcinoma, Merkel cell carcinoma, mucosa-associated lymphoid tissue lymphoma, orbital lymphoma, orbital sarcoma, orbital and optic nerve meningioma, metastatic orbital tumor, lacrimal gland lymphoma, adenoid cystic carcinoma, pleomorphic adenoma, transitional cell carcinoma, and lacrimal sac lymphoma); fallopian tube cancer (including endometrioid adenocarcinoma, serous adenocarcinoma, and pleomorphic adenoma, transitional cell carcinoma, and lacrimal sac lymphoma) Hodgkin lymphoma (including classical Hodgkin lymphoma, nodular sclerosing Hodgkin lymphoma, lymphocyte-rich classical Hodgkin lymphoma, mixed cytology Hodgkin lymphoma, lymphocyte-reduced Hodgkin lymphoma, lymphocyte-predominant Hodgkin lymphoma), implant-associated anaplastic large cell lymphoma (ALCL); inflammatory breast cancer (IBC); kidney cancer (including renal cell carcinoma, urothelial carcinoma of the kidney, renal pelvis, and ureter); leukemia (acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphoblastic leukemia, chronic myeloid leukemia, Myeloid leukemia (including hepatocellular carcinoma, fibrolamellar hepatocellular carcinoma, angiosarcoma, hepatoblastoma, and angiosarcoma); lung cancer (including non-small cell lung cancer, adenocarcinoma, squamous cell carcinoma, large cell carcinoma, small cell lung cancer, carcinoid tumor, salivary gland carcinoma, pulmonary metastasis, and sarcoma); medulloblastoma; melanoma (including cutaneous melanoma, superficial spreading melanoma, nodular melanoma, lentigo maligna melanoma, acral lentiginous melanoma, ocular melanoma, and mucosal melanoma); mesothelioma (including sarcomatoid mesothelioma and biphasic mesothelioma); multiple endocrine neoplasia (MEN), (multiple endocrine neoplasia syndrome 1) type, including multiple endocrine neoplasia type 2); multiple myeloma; myelodysplastic syndromes (MDS) (including refractory anemia, refractory cytopenia with multilineage dysplasia, refractory anemia with ringed sideroblasts, refractory anemia with excess blasts, and refractory cytopenia with multilineage dysplasia and ringed sideroblasts); myeloproliferative disorders (MPDs) (including polycythemia vera, primary myelofibrosis, essential thrombocythemia, systemic mastocytosis, and hypereosinophilic syndrome); neuroblastoma; neurofibromatosis (including neurofibromatosis type 1, neurofibromatosis type 2, and schwannoma);Non-Hodgkin's lymphoma (including B-cell lymphoma, T-cell lymphoma, NK-cell lymphoma, mucosa-associated lymphoid tissue lymphoma, follicular lymphoma, mantle cell lymphoma, diffuse large cell lymphoma, primary mediastinal large cell lymphoma, anaplastic large cell lymphoma, Burkitt lymphoma, lymphoblastic lymphoma, and marginal zone lymphoma); oral cancer (including squamous cell carcinoma); ovarian cancer (including ovarian epithelial carcinoma, germ cell ovarian carcinoma, stromal ovarian carcinoma, and primary peritoneal ovarian carcinoma); pancreatic cancer (including islet cell carcinoma, sarcoma, lymphoma, pseudopapillary neoplasm, ampullary carcinoma, pancreatoblastoma, and adenocarcinoma) Parathyroid diseases (including hyperparathyroidism, hypoparathyroidism, and parathyroid carcinoma), penile cancer (including squamous cell carcinoma, Kaposi's sarcoma, adenocarcinoma, melanoma, and basal cell carcinoma); pituitary tumors (including non-functioning carcinoma, functioning tumors, and pituitary carcinoma), prostate cancer (including adenocarcinoma and prostatic intraepithelial neoplasia), colon cancer (including adenocarcinoma), retinoblastoma (including unilateral retinoblastoma, bilateral retinoblastoma, and PNET-related retinoblastoma), skin cancer (including basal cell carcinoma, squamous cell carcinoma, and actinic keratosis); skull base tumors (meningioma, pituitary adenoma, acoustic neuroma, and granuloma) soft tissue sarcomas; spinal cord tumors (including intramedullary spinal cord tumors, intradural extramedullary spinal cord tumors, epidural spinal cord tumors, osteoblastomas, enchondroma, aneurysmal bone cysts, giant cell tumors, hemangiomas, eosinophilic granulomas, sarcomas, chordoma, chondrosarcomas, and plasmacytomas); gastric cancer (including lymphomas, gastrointestinal stromal tumors, and carcinoid tumors); testicular cancer (including germ cell tumors, non-seminomas, including seminoma, embryonal carcinoma, yolk sac tumor, teratoma, Sertoli cell tumor, choriocarcinoma, stromal tumor, and Leydig tumor; throat cancer (including squamous cell carcinoma); thyroid cancer (including papillary thyroid carcinoma, follicular thyroid carcinoma, eosinophilic cell carcinoma, medullary thyroid carcinoma, and histopathological thyroid carcinoma); uterine cancer (including endometrioid adenocarcinoma, uterine carcinosarcoma, and uterine sarcoma); vaginal cancer (including squamous cell carcinoma, adenocarcinoma, melanoma, and sarcoma); vulvar cancer (including squamous cell carcinoma, adenocarcinoma, melanoma, and sarcoma); von Hippel-Lindau disease; Waldenstrom hypergammaglobulinemia;and Wilms' tumor;

[0125] In some embodiments, the subject suffers from or has a malignant or non-malignant neoplasm described in items 1-10 below.

[0126] Connective tissues (including adult fibrous tissue, embryonic (myxomatous) fibrous tissue, fat, cartilage, bone, and notochord): 1a. Benign neoplasms of connective tissue include fibroma, myxoma, lipoma, chondroma, osteoma, and fibrous histocytoma. 1b. Malignant neoplasms of connective tissue include fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteosarcoma, chordoma, and malignant fibrous histiocytoma.

[0127] Endothelium and mesothelium (including blood vessels, lymphatic vessels, and mesothelium): 2a. Benign neoplasms of the endothelium and mesothelium include hemangiomas, hemangiopericytomas, and lymphangiomas. 2b. Malignant neoplasms of the endothelium and mesothelium include hemangiosarcoma, angiosarcoma, lymphangiosarcoma, and mesothelioma.

[0128] Blood and lymphoid cells (including hematopoietic cells and lymphoid tissue): 3a. Benign neoplasms of blood and lymphoid cells include preleukemias, myeloproliferative disorders, and plasmacytosis. 3b. Malignant neoplasms of blood and lymphoid cells include plasmacytoma, multiple myeloma, Hodgkin's lymphoma, and non-Hodgkin's lymphoma.

[0129] Muscles (including smooth and striated muscles): 4a. Benign neoplasms of muscle include leiomyomas and rhabdomyomas. 4b. Malignant neoplasms of muscle include leiomyosarcoma and rhabdomyosarcoma.

[0130] Epithelial tissues (including stratified squamous, glandular epithelium (including liver, kidney, and bile duct), transitional epithelium, placenta, and testis): 5a. Benign neoplasms of epithelial tissue include papillomas, seborrheic keratoses and adnexal tumors, adenomas (including hepatic, renal tubular, and bile duct adenomas), transitional cell papillomas, and hydatidiform moles. 5b. Malignant neoplasms of epithelial tissue include squamous cell carcinoma, epidermoid carcinoma, malignant skin adnexal tumor, adenocarcinoma, hepatocellular carcinoma, renal cell carcinoma, Grawitz tumor, cholangiocarcinoma, transitional cell carcinoma, choriocarcinoma, seminoma, and embryonal cell carcinoma.

[0131] Neural tissue (including glial cells, neurons, meninges, and nerve sheaths): 6a. Benign neoplasms of nervous tissue include ganglioneuroma, meningioma, schwannoma, schwannoma, and neurofibroma. 6b. Malignant neoplasms of nervous tissue include glioma (grades I-III), undifferentiated glioblastoma multiforme (grade IV), neuroblastoma, medulloblastoma, malignant meningioma, malignant schwannoma, and neurofibrosarcoma.

[0132] Amine precursor uptake and decarboxylation (APUD) tissues (including pituitary gland, parathyroid gland, thyroid gland (C cells), bronchial lining (Kulchytsky cells), adrenal medulla, pheochromocytoma, pancreas, stomach and intestine, carotid body and chemoreceptor system): 7a. Benign neoplasms of APUD tissue include basophilic adenoma, eosinophilic adenoma, chromophobe adenoma, parathyroid adenoma, C-cell hyperplasia, pheochromocytoma, islet cell adenoma, insulinoma, gastrinoma, carcinoid, nonchromaffin paraneuroma, and paraganglioma. 7b. Malignant neoplasms of APUD tissues include parathyroid carcinoma, medullary thyroid carcinoma, bronchial carcinoid, oat cell carcinoma, malignant pheochromocytoma, pancreatic islet cell carcinoma, malignant carcinoid, malignant carcinoid, and malignant paraganglioma.

[0133] Other neural crest-derived cells (including pigment-producing cells in the skin, eye, and occasionally other sites, Schwann cells of the peripheral nervous system, and Merkel cells in squamous epithelia): 8a. Benign neoplasms of neural crest-derived cells include nevi, schwannomas, and schwannomas. 8b. Malignant neoplasms of neural crest-derived cells include melanoma, malignant schwannoma, and Merkel cell neoplasm.

[0134] Tumors (including breast tissue and renal primordium tissue) 9a. Benign neoplasms of breast tissue include fibroadenomas. 9b. Malignant neoplasms of breast tissue include cystosarcoma phyllodes, and malignant neoplasms of the renal primordium include Wilms tumor.

[0135] Ovaries and testes (including multipotent cells, germ cells, connective tissue stroma, and epithelial tissue) 10a. Malignant neoplasms of the ovary and testis include seminoma (in females, dysgerminoma), choriocarcinoma, embryonal carcinoma, endodermal sinus tumor, as well as teratocarcinoma and Sertoli-Leydig cell tumor.

[0136] In some embodiments, the subject has or has a non-malignant tumor selected from the group consisting of adenoma (including polyps of the liver, adrenal gland, pituitary gland, thyroid, and colon and stomach), astrocytoma, craniopharyngioma, desmoid, enchondroma, fibroma or fibroid of any organ such as uterine fibroid, ganglioneuroma, hemangioma, lipoma, lipoblastoma, pheochromocytoma, lymphangioma, meningioma, fibroid (including leiomyoma and rhabdomyoma), nevi or mole, neuroma, neurofibroma, schwannoma, osteochondroma, and papilloma (including skin, cervix, breast duct, and conjunctiva).

[0137] In some embodiments, the subject is a cancer patient, and the cancer may be breast cancer, melanoma, colon cancer, urinary cancer, lung cancer, small cell and non-small cell lung cancer, relapsed or refractory malignancies, non-Hodgkin's lymphoma and Hodgkin's lymphoma, lymphoma, follicular lymphoma, lymphocytic lymphoma, CNS lymphoma, T-cell lymphoma, AIDS-related lymphoma, acute lymphoblastic leukemia, gastrointestinal cancer, liver cancer, hepatocellular carcinoma, ovarian cancer, pancreatic cancer, bile duct cancer, prostate cancer, renal cancer, bladder cancer, colorectal cancer, multiple myeloma, mesothelioma, cervical cancer, vaginal cancer, anal cancer, oropharyngeal cancer, bone marrow cancer, Myeloid leukemia, chronic myeloid leukemia, gastric cancer, nasopharyngeal carcinoma, head and neck carcinoma, glioblastoma, gliosarcoma, squamous cell brain cancer, malignant glioma, diffuse pontine glioma, esophageal cancer, thyroid cancer, astrocytoma, breast cancer, endometrial cancer, skin cell carcinoma, leukemia, acinic cell carcinoma, adenocarcinoma, bronchioloalveolar carcinoma, cholangiocellular carcinoma, chordoma, giant cell carcinoma, intestinal carcinoma, major salivary gland carcinoma, malignant odontogenic neoplasm, malignant peripheral nerve sheath tumor, skin cancer, testicular cancer, germ cell tumor, neuroendocrine carcinoma, parathyroid carcinoma, pituitary carcinoma, placental choriocarcinoma, scrotal cancer, tracheal cancer, transitional cell carcinoma, uterine cancer Cancer of the uterine tract, vulva, kidney, rectum, fallopian tube, peritoneal, epithelial, pleural mesothelioma, sarcomatoid carcinoma, synovial sarcoma, nephroblastoma, neuroblastoma, adult acute myeloid leukemia, myelodysplastic / myeloproliferative neoplasms, embryonal carcinoma, Kaposi's sarcoma, bone cancer, uterine, stomach, endometrial, small intestine, endocrine system, paragland, adrenal, soft tissue sarcoma, urethral, ​​penile, ureteral, pelvis, central nervous system, primary tumor angiogenesis, spinal axis tumors, epidermoid carcinoma, asbestos-induced Environmentally induced cancers including originating cancers, adenosarcoma, adenosquamous carcinoma, adrenocortical carcinoma, astrocytic tumor, basal cell carcinoma, chondrosarcoma, Ewing's sarcoma, gallbladder cancer, hypopharyngeal cancer, intraocular melanoma, laryngeal cancer, leiomyosarcoma, lip and oral cavity cancer, malignant mesothelial tumor, malignant thymoma, medulloblastoma, medulloepithelioma, Merkel cell carcinoma, mucinous epithelioid carcinoma, myelodysplastic syndrome, nasal cavity and paranasal sinus cancer, osteosarcoma, pulmonary blastoma, pineal and supratentorial primitive neuroectodermal tumor, plasma cell neoplasm, retinoblastoma, rhabdomyosarcoma, sarcoma, neuroectodermal tumor,and Wilms' tumor.

[0138] In some embodiments, the subject is a cancer patient suffering from or having a solid tumor. In some embodiments, the cancer patient has a solid tumor infiltrated with lymphocyte cells that express GPR174.

[0139] In some embodiments, the subject has one or more tumors infiltrated with regulatory T cells, such as, for example, breast cancer, lung cancer (such as small cell lung cancer or non-small cell lung cancer), colorectal cancer, cervical cancer, renal cancer, ovarian cancer, melanoma, pancreatic cancer, hepatocellular carcinoma, gastric cancer, glioblastoma, glioma, bladder cancer, myeloma (such as multiple myeloma), prostate cancer, thyroid cancer, testicular cancer, and esophageal cancer.

[0140] In some embodiments, the method comprises administering a GPR174 inhibitory compound as a single agent. In some embodiments, the method comprises administering a GPR174 inhibitory compound in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent. In some embodiments, the method comprises administering a GPR174 inhibitory compound in combination with an A2aR antagonist. In some embodiments, the method comprises administering a GPR174 inhibitory compound in combination with an A2bR antagonist. In some embodiments, the method includes administering a GPR174 inhibitory compound in combination with a Treg-attenuating agent, such as an agent that binds to one or more of GITR, CTLA-4, CD25, LAG3, TIGIT, NRP1, TGF-β, CCR2, CCR4, CCR8, TNFR2, and / or EZH2 to attenuate, deplete, or otherwise reduce the tumor-suppressive function of Treg cells. In some embodiments, the Treg-attenuating agent is a small molecule. In some embodiments, the Treg-attenuating agent is selected from the group consisting of anti-GITR, anti-CTLA-4, anti-CD25, anti-LAG3, anti-TIGIT, anti-NRP1, anti-TGF-β, anti-CCR2, anti-CCR4, anti-CCR8, anti-TNFR2, and / or anti-EZH2 antibodies. Exemplary Treg-attenuating agents for use in the compositions and methods of the present disclosure are provided in Example 25.

[0141] In some embodiments, the method further comprises administering one or more additional therapeutic agents. In certain embodiments, the cancer cells are further contacted with a chemotherapeutic agent.

[0142] In another aspect, the present disclosure features a pharmaceutical composition comprising a combination of a GPR174 inhibitor selected from any of the compounds described below (e.g., those of Formulas I-VI and compounds 1-59 in Table 1) and an inhibitor of ATP-adenosine-A2aR-A2bR-mediated signaling (such as an A2aR antagonist, an A2bR antagonist, and / or a CD73 inhibitor, and / or a CD38 inhibitor, and / or a CD39 inhibitor, and / or a Treg attenuating agent), and a pharmaceutically acceptable carrier. The pharmaceutical composition may be in unit dosage form. In certain embodiments, the compound may be formulated for oral, intravenous, intraperitoneal, intramuscular, topical, rectal, cutaneous, subcutaneous, nasal, cutaneous (e.g., as a transdermal patch), intracerebroventricular, intraparenchymal, intrathecal, inhalation, intracranial, or ocular administration. The pharmaceutical composition may be used to practice the methods disclosed herein.

[0143] In another aspect, the disclosure features compounds of Formula I that have activity as GPR174 inhibitors. In some embodiments, the compounds of Formula I have a structure according to Formula IA or Formula IB or Formula IC described herein. In some embodiments, the disclosure features pharmaceutical compositions including compounds of Formula I, such as compounds according to Formula IA or Formula IB or Formula IC described herein. Compounds of Formula I are useful in methods of inhibiting at least one GPR174-mediated signaling pathway as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuator, to treat or prevent cancers described herein.

[0144] In another aspect, the present disclosure features a compound of Formula II having activity as a GPR174 inhibitor. In some embodiments, the compound of Formula II has a structure according to Formula IIA, Formula IIB, Formula IIC, Formula IID, or Formula IIE described herein. In some embodiments, the present disclosure features a pharmaceutical composition comprising a compound of Formula II, such as a compound according to Formula IIA, Formula IIB, Formula IIC, Formula IID, or Formula IIE described herein. The compound of Formula II is useful in methods of inhibiting at least one GPR174-mediated signaling pathway as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuator, for treating or preventing the cancers described herein.

[0145] In another aspect, the disclosure features a compound of Formula III having activity as a GPR174 inhibitor. In some embodiments, the compound of Formula III has a structure according to Formula IIIA, Formula IIIB, Formula IIIC, Formula IIID, Formula IIIE, or Formula IIIF described herein. In some embodiments, the disclosure features a pharmaceutical composition comprising a compound of Formula III, such as a compound according to Formula IIIA, Formula IIIB, Formula IIIC, Formula IIID, Formula IIIE, or Formula IIIF described herein. The compound of Formula III is useful in methods of inhibiting at least one GPR174-mediated signaling pathway as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuator, for treating or preventing cancers described herein.

[0146] In another aspect, the present disclosure features a compound of formula IV having activity as a GPR174 inhibitor. In some embodiments, the compound of formula IV has a structure according to formula IVA, formula IVB, formula IVC, formula IVD, or formula IVE described herein. In some embodiments, the present disclosure features a pharmaceutical composition comprising a compound of formula IV, such as a compound according to formula IVA, formula IVB, formula IVC, formula IVD, or formula IVE described herein. The compound of formula IV is useful in methods of inhibiting at least one GPR174-mediated signaling pathway as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuator, for treating or preventing cancers described herein.

[0147] In another aspect, the disclosure features compounds of Formula V having activity as GPR174 inhibitors. In some embodiments, the disclosure features pharmaceutical compositions including compounds of Formula V described herein. Compounds of Formula V are useful in methods of inhibiting at least one GPR174-mediated signaling pathway as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuator to treat or prevent the cancers described herein.

[0148] In another aspect, the present disclosure features a compound of formula Va having activity as a GPR174 inhibitor. In some embodiments, the present disclosure features a pharmaceutical composition comprising a compound of formula Va described herein. The compound of formula Va is useful in methods of inhibiting at least one GPR174-mediated signaling pathway, either as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuator, to treat or prevent the cancers described herein.

[0149] In another aspect, the present disclosure features a compound of formula VI having activity as a GPR174 inhibitor. In some embodiments, the present disclosure features a pharmaceutical composition comprising a compound of formula VI described herein. The compound of formula VI is useful in a method for inhibiting at least one GPR174-mediated signaling pathway, either as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuator, for treating or preventing the cancers described herein.

[0150] In another aspect, the disclosure provides a method for preparing a pharmaceutical composition comprising a compound that is an inhibitor of a GPR174-mediated signaling pathway, wherein the compound is contacted in vitro with a mammalian cell that expresses GPR174 and determined to inhibit the GPR174-mediated signaling pathway, wherein the ability of the compound to inhibit the GPR174-mediated signaling pathway indicates that the compound is a GPR174 inhibitor, the method comprising formulating the GPR174 inhibitory compound with a pharmaceutically acceptable carrier as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, wherein the GPR174 inhibitory compound meets the following criteria: (i) the compound has a structure selected from the group consisting of Formulas I-VI; (ii) the compound alters the binding affinity of any one of Reference Compounds 1-59 (listed in Table 1) to GPR174; and / or (iii) the compound differentially inhibits the inhibitory activity of any one of Reference Compounds 1-59 (listed in Table 1) in a GPR174-mediated signaling pathway assay compared to the inhibitory activity of the reference compound alone when tested in the presence of said reference compound: The method is characterized by at least one of:

[0151] In another aspect, the disclosure provides a method for preparing a pharmaceutical composition comprising a compound that is an inhibitor of a GPR174-mediated signaling pathway, wherein the compound has been contacted in vitro with a mammalian cell expressing GPR174 and determined to inhibit the GPR174-mediated signaling pathway, wherein the ability of the compound to inhibit the GPR174-mediated signaling pathway indicates that the compound is a GPR174 inhibitor, the method comprising formulating the GPR174 inhibitory compound with a pharmaceutically acceptable carrier as a single agent or in combination with at least one of an A2aR antagonist, an A2bR antagonist, a CD38 inhibitor, a CD39 inhibitor, a CD73 inhibitor, and / or a Treg attenuating agent, wherein the GPR174 inhibitory compound meets the following criteria: (i) the compound has a structure selected from the group consisting of Formulas I-VI; (ii) the compound alters the apparent binding affinity of any one of Reference Compounds 1-59 (listed in Table 1) to GPR174; and / or (iii) the compound differentially inhibits the inhibitory activity of any one of Reference Compounds 1-59 (listed in Table 1) in a GPR174-mediated signaling pathway assay compared to the inhibitory activity of the reference compound alone when tested in the presence of said reference compound: The method is characterized by at least one of:

[0152] In another aspect, the disclosure features a GPR174-interacting compound that includes a structure according to any of Formulas I-VI disclosed herein and a detectable moiety. In some aspects, the GPR174-interacting compound meets the following criteria: (i) the compound has a structure selected from the group consisting of Formulas I-VI; (ii) the compound alters the binding affinity of any one of Reference Compounds 1-59 (listed in Table 1) to GPR174; and / or (iii) the compound differentially inhibits the inhibitory activity of any one of Reference Compounds 1-59 (listed in Table 1) in a GPR174-mediated signaling pathway assay compared to the inhibitory activity of the reference compound alone when tested in the presence of said reference compound: In one embodiment, the GPR174-interacting compound comprising a detectable moiety comprises a structure according to any of compounds 1-59 disclosed in Table 1. In certain embodiments, the detectable moiety is at least one of a radioisotope, a bioluminescent tag, a chemiluminescent tag, or a photoaffinity label. In one embodiment, the detectable moiety is a radioisotope.

[0153] In another aspect, the disclosure features a GPR174-interacting compound that includes a structure according to any of Formulas I-VI disclosed herein and a detectable moiety. In some aspects, the GPR174-interacting compound meets the following criteria: (i) the compound has a structure selected from the group consisting of Formulas I-VI; (ii) the compound alters the apparent binding affinity of any one of Reference Compounds 1-59 (listed in Table 1) to GPR174; and / or (iii) the compound differentially inhibits the inhibitory activity of any one of Reference Compounds 1-59 (listed in Table 1) in a GPR174-mediated signaling pathway assay compared to the inhibitory activity of the reference compound alone when tested in the presence of said reference compound: In one embodiment, the GPR174-interacting compound comprising a detectable moiety comprises a structure according to any of compounds 1-59 disclosed in Table 1. In certain embodiments, the detectable moiety is at least one of a radioisotope, a bioluminescent tag, a chemiluminescent tag, or a photoaffinity label. In one embodiment, the detectable moiety is a radioisotope.

[0154] In any of the above aspects, the GPR174 inhibitory compound can be a synthetic compound, a semi-synthetic compound (e.g., a chemically modified steroid derived from a natural source), or a natural product. In certain embodiments, the compound is not an endogenous ligand of GPR174 (e.g., a surrogate ligand). In various embodiments, the compound is a functional inhibitor of a GPR174-mediated signal transduction pathway (e.g., an antagonist, partial agonist, inverse agonist, partial inverse agonist, or negative allosteric modulator). In some embodiments, the compound is an antagonist of GPR174 and inhibits at least one GPR174-mediated signal transduction pathway in the presence of an agonist or partial agonist. In some embodiments, the compound is an inverse agonist. In certain embodiments, the compound reduces activation of a GPR174-mediated signaling pathway by at least two-thirds, one-half, one-fifth, one-tenth, one-fifth, or one-hundredth of basal activity at concentrations less than 40, 25, 10, 5, 1, 0.5, 0.25, 0.1, or 0.5 μM (e.g., 0.01 μM to 0.5 μM, 0.01 μM to 0.5 μM, 0.01 μM to 5 μM, 0.01 μM to 10 μM, 0.01 μM to 25 μM, or 0.01 μM to 40 μM). In some embodiments, the compound is an inverse agonist and reduces activation of at least one GPR174-mediated signaling pathway by at least two-fold compared to basal activity at concentrations less than 10 μM (e.g., 0.01 μM to 10 μM).

[0155] The GPR174 inhibitory compound may be a polypeptide, an antibody, a non-peptide compound, an expression inhibitor that inhibits GPR174 expression (e.g., a GPR174 antisense nucleic acid molecule such as an antisense RNA, antisense DNA or antisense oligonucleotide, a GPR174 ribozyme or a GPR174 RNAi molecule), or a small molecule (e.g., a small organic molecule or organometallic molecule).

[0156] In some embodiments, the compound was not under development by any entity before January 11, 2019. In some embodiments, the compound is not a compound approved by a regulatory agency. In some embodiments, the compound is a compound that was not approved by the U.S. Food and Drug Administration or other regulatory agency for use in diseases associated with GPR174 before January 11, 2019. In some embodiments, the compound was not under development by any entity for use in diseases associated with GPR174 before January 11, 2019. In some embodiments, the compound was not approved by the U.S. Food and Drug Administration or equivalent regulatory agency as targeting GPR174 before January 11, 2019. In certain embodiments, the compound is not an antibody or receptor-binding fragment thereof. In certain embodiments, the compound is not a pepducin, which is a membrane-tethered peptide that penetrates cells (see, e.g., Covic L. et al., PNAS 99(2):643-8(2002)).

[0157] In particular embodiments of any aspect, the isolated GPR174 inhibitory compound is not LysoPS or PS.

[0158] In certain embodiments, the GPR174 inhibitory compound can functionally interact with human GPR174 having the sequence of SEQ ID NO:1 and / or can alter the cellular distribution of a recombinant GPR174 polypeptide (e.g., human GPR174 having the sequence of SEQ ID NO:1) modified to include a nuclear localization signal compared to the cellular distribution of the recombinant GPR174 polypeptide in the absence of the compound.

[0159] The compound may be specific for GPR174, for example, as compared to a reference panel of GPCRs.

[0160] In particular embodiments of any aspect, the isolated compound selectively binds to GPR174 and does not bind to P2Y10.

[0161] In particular embodiments of any aspect, the isolated compound selectively binds to GPR174 and does not bind to GPR34.

[0162] In particular embodiments of any aspect, the isolated compound functionally interacts with GPR174 and does not bind to P2Y10.

[0163] In particular embodiments of any aspect, the isolated compound functionally interacts with GPR174 and does not bind to GPR34.

[0164] In certain embodiments of any aspect, the isolated compound is not disclosed in US2015 / 0361119.

[0165] Additionally, the methods and compositions of any of the aspects of the disclosure may employ any compound having a structure according to the formula set forth below, such as the exemplary compounds in Table 1, or a pharmaceutically acceptable salt thereof.

[0166] In some embodiments, the GPR174 inhibitory compound has the following formula (I): TIFF0007625524000001.tif48128 or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof; During the ceremony, X 1 is N or CR 10 and; X 2 is N or CR 11 and; X 3 is N or CR 12 and; X 4 is N or CR 13 and; X 5 is N or CR 14 and; X 6 is N or CR 15 and; X 7 is N or CR 16 and; R1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , and R 9 each independently represents H, hydroxy, thiol, optionally substituted amino, optionally substituted amido, cyano, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted C1-C6 alkoxy, optionally substituted C6-C 10 Aryloxy, optionally substituted C1-C9 heteroaryloxy, optionally substituted C2-C6 alkanoyl, optionally substituted C7-C 11 Aryloyl, optionally substituted C2-C 10 Heteroaryloyl, optionally substituted C2-C 10 Heterocyclyl, hydroxycarbonyl, optionally substituted ester, optionally substituted carboxamide, optionally substituted C1-C6 alkanoyloxy, optionally substituted C7-C 11 Aryloyloxy, optionally substituted C2-C 10 Heteroaryloyloxy, optionally substituted C2-C 10 Heterocyclyloxy, optionally substituted C1-C6 alkylsulfinyl, optionally substituted C1-C6 alkylsulfonyl, optionally substituted C6-C 10 Arylsulfinyl, optionally substituted C6-C 10 Arylsulfonyl, optionally substituted C1-C9 heteroarylsulfinyl, optionally substituted C1-C9 heteroarylsulfonyl, optionally substituted C1-C9 heterocyclylsulfinyl, optionally substituted C1-C9 heterocyclylsulfonyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C2-C6 heteroalkenyl, optionally substituted C2-C6 heteroalkynyl, optionally substituted C3-C 10Cycloalkyl, optionally substituted C4-C 10 Cycloalkenyl, optionally substituted C8-C 10 Cycloalkynyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10 Aryl C1-C6 alkyl, optionally substituted C6-C 10 Aryl C2-C6 alkenyl, optionally substituted C6-C 10 aryl C2-C6 alkynyl, optionally substituted C1-C9 heteroaryl, optionally substituted C1-C9 heteroaryl C1-C6 alkyl, optionally substituted C1-C9 heteroaryl C2-C6 alkenyl, optionally substituted C1-C9 heteroaryl C2-C6 alkynyl, optionally substituted C1-C9 heterocyclyl, optionally substituted C1-C9 heterocyclyl C1-C6 alkyl, optionally substituted C1-C9 heterocyclyl C2-C6 alkenyl, or optionally substituted C1-C9 heterocyclyl C2-C6 alkynyl; or R 2 and R 3 is bonded to =O, =S, or =NR 17 or R 4 and R 5 is bonded to =O, =S, or =NR 17 or R 6 and R 7 is bonded to =O, =S, or =NR 17 or R 8 and R 9 is bonded to =O, =S, or =NR 17 Forming; R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , and R 16each independently represents H, hydroxy, halogen, thiol, optionally substituted amino, optionally substituted amido, cyano, nitro, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted C1-C6 alkoxy, optionally substituted C6-C 10 Aryloxy, optionally substituted C1-C9 heteroaryloxy, optionally substituted C2-C6 alkanoyl, optionally substituted C7-C 11 Aryloyl, optionally substituted C2-C 10 Heteroaryloyl, optionally substituted C2-C 10 Heterocyclyl, hydroxycarbonyl, optionally substituted ester, optionally substituted carboxamide, optionally substituted C1-C6 alkanoyloxy, optionally substituted C7-C 11 Aryloyloxy, optionally substituted C2-C 10 Heteroaryloyloxy, optionally substituted C2-C 10 Heterocyclyloxy, optionally substituted C1-C6 alkylsulfinyl, optionally substituted C1-C6 alkylsulfonyl, optionally substituted C6-C 10 Arylsulfinyl, optionally substituted C6-C 10 Arylsulfonyl, optionally substituted C1-C9 heteroarylsulfinyl, optionally substituted C1-C9 heteroarylsulfonyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C2-C6 heteroalkenyl, optionally substituted C2-C6 heteroalkynyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C4-C 10 Cycloalkenyl, optionally substituted C8-C 10 Cycloalkynyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10 Aryl C1-C6 alkyl, optionally substituted C6-C 10Aryl C2-C6 alkenyl, optionally substituted C6-C 10 aryl C2-C6 alkynyl, optionally substituted C1-C9 heteroaryl, optionally substituted C1-C9 heteroaryl C1-C6 alkyl, optionally substituted C1-C9 heteroaryl C2-C6 alkenyl, optionally substituted C1-C9 heteroaryl C2-C6 alkynyl, optionally substituted C1-C9 heterocyclyl, optionally substituted C1-C9 heterocyclyl C1-C6 alkyl, optionally substituted C1-C9 heterocyclyl C2-C6 alkenyl, or optionally substituted C1-C9 heterocyclyl C2-C6 alkynyl; or (i)R 12 and R 13 are joined together with the atoms to which they are each attached to form an optionally substituted 5-, 6-, or 7-membered ring; (ii)R 13 and R 14 are joined together with the atoms to which they are each attached to form an optionally substituted 5-, 6-, or 7-membered ring; (iii)R 14 and R 15 are joined together with the atoms to which they are each attached to form an optionally substituted 5-, 6-, or 7-membered ring; and (iv)R 15 and R 16 may be joined together with the atoms to which they are attached to form an optionally substituted 5-, 6-, or 7-membered ring. is one of R 17 is H, hydroxyl, cyano, optionally substituted amino, optionally substituted amido, optionally substituted carboxamido, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted C1-C6 alkanoyl, optionally substituted C7-C 11 Aryloyl, optionally substituted C2-C 10Heterocyclyl, optionally substituted C2-C 10 Heteroaryloyl, optionally substituted C1-C6 alkylsulfinyl, optionally substituted C1-C6 alkylsulfonyl, optionally substituted C6-C 10 Arylsulfinyl, optionally substituted C6-C 10 Arylsulfonyl, optionally substituted C1-C9 heteroarylsulfinyl, optionally substituted C1-C9 heteroarylsulfonyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C2-C6 heteroalkenyl, optionally substituted C2-C6 heteroalkynyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C4-C 10 Cycloalkenyl, optionally substituted C8-C 10 Cycloalkynyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10 Aryl C1-C6 alkyl, optionally substituted C6-C 10 Aryl C2-C6 alkenyl, optionally substituted C6-C 10 aryl C2-C6 alkynyl, optionally substituted C1-C9 heteroaryl, optionally substituted C1-C9 heteroaryl C1-C6 alkyl, optionally substituted C1-C9 heteroaryl C2-C6 alkenyl, optionally substituted C1-C9 heteroaryl C2-C6 alkynyl, optionally substituted C1-C9 heterocyclyl, optionally substituted C1-C9 heterocyclyl C1-C6 alkyl, optionally substituted C1-C9 heterocyclyl C2-C6 alkenyl, or optionally substituted C1-C9 heterocyclyl C2-C6 alkynyl; X 3 , X 4 , X 5 , X 6 , and X 7 three or fewer of are N; and X 1 and X 2 At least one of is N.

[0167] In some embodiments of Formula (I), X 1 is N. In certain embodiments of formula (I), X 2 is N. In certain embodiments of formula (I), X 3 is CR 12 In another embodiment of formula (I), X 4 is CR 13 In yet another embodiment of formula (I), X 5 is CR 14 In yet another embodiment of formula (I), X 6 is CR 15 In certain other embodiments of formula (I), X 7 is CR 16 is.

[0168] In certain embodiments of Formula (I), the isolated compound has the formula (IA): It has the structure according to TIFF0007625524000002.tif39128.

[0169] In certain embodiments of formula (I) or (IA), R 2 is H, optionally substituted C1-C6 alkyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10 In some embodiments of Formula (I) or (IA), R is an aryl C1-C6 alkyl, an optionally substituted C1-C9 heteroaryl, an optionally substituted C1-C9 heteroaryl C1-C6 alkyl, an optionally substituted C1-C9 heterocyclyl, or an optionally substituted C1-C9 heterocyclyl C1-C6 alkyl. 2 is H or optionally substituted C1-C6 alkyl. In other embodiments of formula (I) or (IA), R 2 is H.

[0170] In yet another embodiment of formula (I) or (IA), R 3 is H, optionally substituted C1-C6 alkyl, optionally substituted C3-C 10Cycloalkyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10 In yet another embodiment of formula (I) or (IA), R is an aryl C1-C6 alkyl, an optionally substituted C1-C9 heteroaryl, an optionally substituted C1-C9 heteroaryl C1-C6 alkyl, an optionally substituted C1-C9 heterocyclyl, or an optionally substituted C1-C9 heterocyclyl C1-C6 alkyl. 3 is H or optionally substituted C1-C6 alkyl. In certain embodiments of formula (I) or (IA), R 3 is H.

[0171] In some embodiments of formula (I) or (IA), R 4 is H, optionally substituted C1-C6 alkyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10 In certain embodiments of formula (I) or (IA), R is an aryl C1-C6 alkyl, an optionally substituted C1-C9 heteroaryl, an optionally substituted C1-C9 heteroaryl C1-C6 alkyl, an optionally substituted C1-C9 heterocyclyl, or an optionally substituted C1-C9 heterocyclyl C1-C6 alkyl. 4 is H or optionally substituted C1-C6 alkyl. In certain embodiments of formula (I) or (IA), R 4 is H.

[0172] In another embodiment of formula (I) or (IA), R 5 is H, optionally substituted C1-C6 alkyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10In yet another embodiment of formula (I) or (IA), R is an aryl C1-C6 alkyl, an optionally substituted C1-C9 heteroaryl, an optionally substituted C1-C9 heteroaryl C1-C6 alkyl, an optionally substituted C1-C9 heterocyclyl, or an optionally substituted C1-C9 heterocyclyl C1-C6 alkyl. 5 is H or optionally substituted C1-C6 alkyl. In still other embodiments of formula (I) or (IA), R 5 is H.

[0173] In certain embodiments of formula (I) or (IA), R 6 is H, optionally substituted C1-C6 alkyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10 In certain embodiments of formula (I) or (IA), R is an aryl C1-C6 alkyl, an optionally substituted C1-C9 heteroaryl, an optionally substituted C1-C9 heteroaryl C1-C6 alkyl, an optionally substituted C1-C9 heterocyclyl, or an optionally substituted C1-C9 heterocyclyl C1-C6 alkyl. 6 is H or an optionally substituted C1-C6 alkyl. In some embodiments of formula (I) or (IA), R 6 is H.

[0174] In certain embodiments of formula (I) or (IA), R 7 is H, optionally substituted C1-C6 alkyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10In another embodiment of formula (I) or (IA), R is an aryl C1-C6 alkyl, an optionally substituted C1-C9 heteroaryl, an optionally substituted C1-C9 heteroaryl C1-C6 alkyl, an optionally substituted C1-C9 heterocyclyl, or an optionally substituted C1-C9 heterocyclyl C1-C6 alkyl. 7 is H or optionally substituted C1-C6 alkyl. In still other embodiments of formula (I) or (IA), R 7 is H.

[0175] In some embodiments of formula (I) or (IA), R 8 is H, optionally substituted C1-C6 alkyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10 In certain embodiments of formula (I) or (IA), R is an aryl C1-C6 alkyl, an optionally substituted C1-C9 heteroaryl, an optionally substituted C1-C9 heteroaryl C1-C6 alkyl, an optionally substituted C1-C9 heterocyclyl, or an optionally substituted C1-C9 heterocyclyl C1-C6 alkyl. 8 is H or optionally substituted C1-C6 alkyl. In certain embodiments of formula (I) or (IA), R 8 is H.

[0176] In another embodiment of formula (I) or (IA), R 9 is H, optionally substituted C1-C6 alkyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10In yet another embodiment of formula (I) or (IA), R is an aryl C1-C6 alkyl, an optionally substituted C1-C9 heteroaryl, an optionally substituted C1-C9 heteroaryl C1-C6 alkyl, an optionally substituted C1-C9 heterocyclyl, or an optionally substituted C1-C9 heterocyclyl C1-C6 alkyl. 9 is H or optionally substituted C1-C6 alkyl. In still other embodiments of formula (I) or (IA), R 9 is H.

[0177] In certain embodiments of formula (I) or (IA), R 13 is H, hydroxy, optionally substituted amino, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C1-C6 alkoxy, optionally substituted C6-C 10 Aryloxy, optionally substituted C1-C9 heteroaryloxy, optionally substituted C6-C 10 aryl, optionally substituted C1-C9 heteroaryl, or optionally substituted C1-C9 heterocyclyl. In certain embodiments of formula (I) or (IA), R 13 is H or optionally substituted C1-C6 alkyl. In other embodiments of formula (I) or (IA), R 13 is H.

[0178] In certain embodiments of formula (I) or (IA), R 16 is H, halogen, cyano, nitro, optionally substituted C1-C6 alkyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C6-C 10 Aryl, optionally substituted C1-C9 heteroaryl, optionally substituted C2-C6 alkanoyl, optionally substituted C7-C 11 Aryloyl, optionally substituted C2-C 10 Heteroaryloyl, optionally substituted C2-C 10In another embodiment of formula (I) or (IA), R is heterocyclyl, hydroxycarbonyl, an optionally substituted ester, or an optionally substituted C1-C9 heterocyclyl. 16 is H or optionally substituted C1-C6 alkyl. In yet another embodiment of formula (I) or (IA), R 16 is H.

[0179] In some embodiments of Formula (I), the compound has the formula (IB): It has the structure according to TIFF0007625524000003.tif39128.

[0180] In certain embodiments of formula (I), (IA), or (IB), R 12 is H, halogen, cyano, nitro, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxy, optionally substituted C1-C6 alkanoyl, optionally substituted C7-C 11 Aryloyl, optionally substituted C2-C 10 Heterocyclyl, optionally substituted C2-C 10 Heteroaryloyl, optionally substituted C1-C6 alkylsulfonyl, optionally substituted C6-C 10 Arylsulfonyl, optionally substituted C1-C9 heteroarylsulfonyl, hydroxycarbonyl, optionally substituted ester, optionally substituted carboxamide, optionally substituted C1-C6 alkanoyloxy, optionally substituted C7-C 11 Aryloyloxy, optionally substituted C2-C 10 Heteroaryloyloxy, optionally substituted C2-C 10 In certain embodiments of formula (I), (IA), or (IB), R is heterocyclyloxy, optionally substituted C1-C9 heteroaryl, or optionally substituted C1-C9 heterocyclyl. 12is H, halogen, cyano, nitro, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxy, optionally substituted C1-C6 alkanoyl, optionally substituted C1-C6 alkylsulfonyl, hydroxycarbonyl, optionally substituted ester, optionally substituted carboxamido, optionally substituted C1-C6 alkanoyloxy, optionally substituted C1-C9 heteroaryl, or optionally substituted C1-C9 heterocyclyl. In other embodiments of Formula (I), (IA), or (IB), R 12 is H, halogen, cyano, nitro, optionally substituted C1-C6 alkyl, optionally substituted ester, optionally substituted carboxamido, optionally substituted C1-C6 alkanoyloxy, or optionally substituted C1-C9 heteroaryl. In still other embodiments of Formula (I), (IA), or (IB), R 12 is H, halogen, nitro, an optionally substituted ester, or an optionally substituted C1-C6 alkanoyloxy. In still other embodiments of Formula (I), (IA), or (IB), R 12 is a halogen (e.g., R 12 is fluorine). In certain embodiments of formula (I), (IA), or (IB), R 12 is nitro.

[0181] In certain embodiments of formula (I), (IA), or (IB), R 14 is H, halogen, cyano, nitro, optionally substituted C1-C6 alkyl, hydroxycarbonyl, optionally substituted ester, optionally substituted carboxamide, optionally substituted C1-C6 alkanoyloxy, optionally substituted C7-C 11 Aryloyloxy, optionally substituted C2-C 10 Heteroaryloyloxy, optionally substituted C2-C 10 Heterocyclyloxy, optionally substituted C1-C6 alkanoyl, optionally substituted C7-C 11 Aryloyl, optionally substituted C2-C 10Heterocyclyl, optionally substituted C2-C 10 Heteroaryloyl, optionally substituted C1-C6 alkylsulfonyl, optionally substituted C6-C 10 Arylsulfonyl, optionally substituted C1-C9 heteroarylsulfinyl, optionally substituted C6-C 10 In some embodiments of formula (I), (IA), or (IB), R is aryl, optionally substituted C1-C9 heteroaryl, or optionally substituted C1-C9 heterocyclyl. 14 is H, halogen, cyano, optionally substituted C1-C6 alkyl, optionally substituted ester, optionally substituted carboxamido, optionally substituted C1-C6 alkanoyloxy, optionally substituted C1-C6 alkanoyl, optionally substituted C7-C 11 Aryloyl, optionally substituted C2-C 10 Heterocyclyl, optionally substituted C2-C 10 Heteroaryloyl, optionally substituted C1-C6 alkylsulfonyl, optionally substituted C6-C 10 In other embodiments of formula (I), (IA), or (IB), R 14 is H, halogen, optionally substituted C1-C6 alkyl, optionally substituted ester, optionally substituted carboxamido, optionally substituted C1-C6 alkanoyloxy, optionally substituted C1-C6 alkanoyl. In still other embodiments of Formula (I), (IA), or (IB), R 14 is halogen, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 alkanoyl. In still other embodiments of Formula (I), (IA), or (IB), R 14 is a halogen (e.g., R 14 is fluorine). In some embodiments of formula (I), (IA), or (IB), R 14 is an optionally substituted C1-C6 alkanoyl. In certain embodiments of formula (I), (IA), or (IB), R14 is an optionally substituted C2-C4 alkanoyl. In certain embodiments of formula (I), (IA), or (IB), R 14 is an unsubstituted C2-C4 alkanoyl.

[0182] In some embodiments of formula (I), (IA), or (IB), R 15 is H, optionally substituted amino, optionally substituted amido, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxy, optionally substituted C6-C 10 Aryloxy, optionally substituted C1-C9 heteroaryloxy, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10 In certain embodiments of formula (I), (IA), or (IB), R is an aryl C1-C6 alkyl, an optionally substituted C1-C9 heteroaryl, an optionally substituted C1-C9 heteroaryl C1-C6 alkyl, an optionally substituted C1-C9 heterocyclyl, or an optionally substituted C1-C9 heterocyclyl C1-C6 alkyl. 15 is H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxy, optionally substituted C6-C 10 Aryloxy, optionally substituted C1-C9 heteroaryloxy, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C6-C 10 aryl, optionally substituted C1-C9 heteroaryl, or optionally substituted C1-C9 heterocyclyl. In certain embodiments of formula (I), (IA), or (IB), R 15 is H or an optionally substituted C1-C9 heterocyclyl. In other embodiments of formula (I), (IA), or (IB), R 15 is H. In still other embodiments of formula (I), (IA), or (IB), R15 is an optionally substituted C1-C9 heterocyclyl (e.g., R 15 is piperidinyl, methyl-substituted piperidinyl or benzpiperidinyl).

[0183] In some embodiments of formula (IB), R 1 C1-C6 alkanoyl, C6-C 10 Aryl, C7-C 11 Allyroyle, C2~C 10 Heteroaryloyl, C2-C7 alkoxycarbonyl, and C6-C 10 arylsulfonyl; R 1 is optionally substituted; R 12 is H, nitro, or halogen; R 14 is C1-C6 alkanoyl or halogen; and R 15 is H or an optionally substituted C1-C9 heterocyclyl.

[0184] In some embodiments of formula (IB), R 1 C1-C6-alkanoyl, C7-C 11 Allyroyle, C2~C 10 Heteroaryloyl, C2-C7 alkoxycarbonyl, and C6-C 10 The group is selected from the group consisting of optionally substituted arylsulfonyl.

[0185] In some embodiments of formula (IB), R 12 is nitro and R 14 is fluoro.

[0186] In some embodiments of formula (IB), R 15 is an optionally substituted piperidin-1-yl or an optionally substituted azepan-1-yl. In some embodiments of Formula (I), (IA), or (IB), the compound has the formula (IC): It has the structure according to TIFF0007625524000004.tif34128, In the formula, R 16 is H or C1-C6 alkyl.

[0187] In some embodiments of formula (IC), R 16 is H or methyl.

[0188] In certain embodiments of formula (I), (IA), (IB), or (IC), R 1 is H, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkanoyl, optionally substituted C7-C 11 Aryloyl, optionally substituted C2-C 10 Heteroaryloyl, optionally substituted C2-C 10 Heterocyclyl, optionally substituted ester, optionally substituted carboxamide, optionally substituted C1-C6 alkylsulfonyl, substituted C6-C 10 Arylsulfonyl, optionally substituted C1-C9 heteroarylsulfonyl, optionally substituted C1-C9 heterocyclylsulfonyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10 In certain embodiments of formula (I), (IA), (IB), or (IC), R is an aryl C1-C6 alkyl, an optionally substituted C1-C9 heteroaryl, an optionally substituted C1-C9 heteroaryl C1-C6 alkyl, an optionally substituted C1-C9 heterocyclyl, or an optionally substituted C1-C9 heterocyclyl C1-C6 alkyl. 1 is an optionally substituted C1-C6 alkyl, an optionally substituted C2-C6 alkanoyl, an optionally substituted C7-C 11 Aryloyl, optionally substituted C2-C 10 Heteroaryloyl, optionally substituted C2-C 10Heterocyclyl, optionally substituted ester, optionally substituted carboxamide, optionally substituted C1-C6 alkylsulfonyl, substituted C6-C 10 Arylsulfonyl, optionally substituted C1-C9 heteroarylsulfonyl, optionally substituted C1-C9 heterocyclylsulfonyl, optionally substituted C6-C 10 arylC1-C6 alkyl, optionally substituted C1-C9 heteroarylC1-C6 alkyl, or optionally substituted C1-C9 heterocyclylC1-C6 alkyl. In certain embodiments of formula (I), (IA), (IB), or (IC), R 1 is an optionally substituted C1-C6 alkyl, an optionally substituted C2-C6 alkanoyl, an optionally substituted C7-C 11 Aryloyl, optionally substituted C2-C 10 Heteroaryloyl, optionally substituted ester, optionally substituted carboxamide, optionally substituted C1-C6 alkylsulfonyl, substituted C6-C 10 Arylsulfonyl, optionally substituted C1-C9 heteroarylsulfonyl, optionally substituted C6-C 10 In some embodiments of formula (I), (IA), (IB), or (IC), R is an aryl C1-C6 alkyl, or an optionally substituted C1-C9 heteroaryl C1-C6 alkyl. 1 is an optionally substituted C1-C6 alkyl, an optionally substituted C2-C6 alkanoyl, an optionally substituted C7-C 11 Aryloyl, optionally substituted ester, optionally substituted C-C 10 Arylsulfonyl or optionally substituted C6-C 10 In other embodiments of formula (I), (IA), (IB), or (IC), R 1 is an optionally substituted C1-C6 alkyl, an optionally substituted C2-C6 alkanoyl, an optionally substituted C7-C 11 Aryloyl, optionally substituted C2-C 10Heteroaryloyl, optionally substituted C6-C 10 In still other embodiments of Formula (I), (IA), (IB), or (IC), R 1 is an optionally substituted C2-C7 alkoxycarbonyl (e.g., methyloxycarbonyl or ethyloxycarbonyl). In still other embodiments of Formula (I), (IA), (IB), or (IC), R 1 is an optionally substituted C2-C6 alkanoyl (e.g., R 1 is acetyl, propanoyl, n-butanoyl, isobutanoyl, or t-pentanoyl. In some embodiments of formula (I), (IA), (IB), or (IC), R 1 is an optionally substituted C7 to C 11 aryloyl (e.g., R 1 is 4-fluorobenzoyl or benzoyl). In other embodiments of formula (I), (IA), (IB), or (IC), R 1 is optionally substituted C2 to C 10 heteroaryloyl (e.g., R 1 is 2-thiophenecarbonyl). In certain embodiments of formula (I), (IA), (IB), or (IC), R 1 is an optionally substituted C6 to C 10 arylsulfonyl (e.g., R 1 is p-tolylsulfonyl or phenylsulfonyl). In other embodiments of formula (I), (IA), (IB), or (IC), R 1 is an optionally substituted C1-C6 alkyl (e.g., R 1 is ethyl or methyl).

[0189] In certain embodiments of Formula (I), (IA), (IB), or (1C), the isolated compound is Compound 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 53: TIFF0007625524000005.tif87143TIFF0007625524000006.tif209143TIFF0007625524000007.tif121141.

[0190] In certain embodiments of formula (IB), the isolated compound is compound 53: TIFF0007625524000008.tif37128.

[0191] In some embodiments, the GPR174 inhibitory compound has formula (II): TIFF0007625524000009.tif27128 or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof; During the ceremony, X 1 is N or CR 2 and; X 2 is N or CR 3 and; R A and R B together with the atoms to which they are attached, combine to form an optionally substituted 5-membered ring, an optionally substituted 6-membered ring, or an optionally substituted 7-membered ring; R 1 is H, halo, hydroxy, optionally substituted amino, optionally substituted amido, thiol, cyano, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted C1-C6 alkoxy, optionally substituted C6-C 10 Aryloxy, optionally substituted C1-C9 heteroaryloxy, optionally substituted C2-C6 alkanoyl, optionally substituted C7-C 11 Aryloyl, optionally substituted C2-C 10 Heteroaryloyl, optionally substituted C2-C 10Heterocyclyl, hydroxycarbonyl, optionally substituted C2-C7 alkoxycarbonyl, optionally substituted carboxamido, optionally substituted C1-C6 alkanoyloxy, optionally substituted C7-C 11 Aryloyloxy, optionally substituted C2-C 10 Heteroaryloyloxy, optionally substituted C2-C 10 Heterocyclyloxy, optionally substituted C1-C6 thioalkyl, optionally substituted C1-C6 alkylsulfinyl, optionally substituted C1-C6 alkylsulfonyl, optionally substituted C6-C 10 Arylthio, optionally substituted C6-C 10 Arylsulfinyl, optionally substituted C6-C 10 Arylsulfonyl, optionally substituted C1-C9 heteroarylthio, optionally substituted C1-C9 heteroarylsulfinyl, optionally substituted C1-C9 heteroarylsulfonyl, optionally substituted C1-C9 heterocyclylsulfinyl, optionally substituted C1-C9 heterocyclylsulfonyl, optionally substituted sulfamoyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C2-C6 heteroalkenyl, optionally substituted C2-C6 heteroalkynyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C4-C 10 Cycloalkenyl, optionally substituted C8-C 10 Cycloalkynyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10 Aryl C1-C6 alkyl, optionally substituted C6-C 10 Aryl C2-C6 alkenyl, optionally substituted C6-C 10aryl C2-C6 alkynyl, optionally substituted C1-C9 heteroaryl, optionally substituted C1-C9 heteroaryl C1-C6 alkyl, optionally substituted C1-C9 heteroaryl C2-C6 alkenyl, optionally substituted C1-C9 heteroaryl C2-C6 alkynyl, optionally substituted C1-C9 heterocyclyl, optionally substituted C1-C9 heterocyclyl C1-C6 alkyl, optionally substituted C1-C9 heterocyclyl C2-C6 alkenyl, or optionally substituted C1-C9 heterocyclyl C2-C6 alkynyl; and Ar 1 is an optionally substituted C6 to C 10 It is aryl, optionally substituted C1-C9 heteroaryl, or optionally substituted C1-C9 heterocyclyl.

[0192] In some embodiments of Formula (II), R A and R B and together with the atoms to which they are each attached, combine to form an optionally substituted carbocyclic ring. In certain embodiments of formula (II), R A and R B and together with the atoms to which they are each attached, combine to form an optionally substituted heterocycle. In certain embodiments of formula (II), R A and R B and together with the atoms to which they are each attached, combine to form an optionally substituted 6-membered ring. In another embodiment of formula (II), R A and R B and together with the atoms to which they are each attached, combine to form an optionally substituted non-aromatic ring. In yet another embodiment of Formula (II), R A and R B together with the atoms to which they are each attached, combine to form an optionally substituted aromatic ring.

[0193] In yet another embodiment of Formula (II), the isolated compound has the formula (IIA): It has the structure according to TIFF0007625524000010.tif34128, During the ceremony, X 3 N, CR 4 and; X 4 N, CR 5 and; X 5 N, CR 6 and; X 6 N, CR 7 , or non-existent; and R 4 , R 5 , R 6 , and R 7 each independently represents H, halo, hydroxy, optionally substituted amino, optionally substituted amido, thiol, cyano, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted C1-C6 alkoxy, optionally substituted C6-C 10 Aryloxy, optionally substituted C1-C9 heteroaryloxy, optionally substituted C2-C6 alkanoyl, optionally substituted C7-C 11 Aryloyl, optionally substituted C2-C 10 Heteroaryloyl, optionally substituted C2-C 10 Heterocyclyl, hydroxycarbonyl, optionally substituted C2-C7 alkoxycarbonyl, optionally substituted carboxamido, optionally substituted C1-C6 alkanoyloxy, optionally substituted C7-C 11 Aryloyloxy, optionally substituted C2-C 10 Heteroaryloyloxy, optionally substituted C2-C 10 Heterocyclyloxy, optionally substituted C1-C6 thioalkyl, optionally substituted C1-C6 alkylsulfinyl, optionally substituted C1-C6 alkylsulfonyl, optionally substituted C6-C 10 Arylthio, optionally substituted C6-C 10 Arylsulfinyl, optionally substituted C6-C 10Arylsulfonyl, optionally substituted C1-C9 heteroarylthio, optionally substituted C1-C9 heteroarylsulfinyl, optionally substituted C1-C9 heteroarylsulfonyl, optionally substituted C1-C9 heterocyclylsulfinyl, optionally substituted C1-C9 heterocyclylsulfonyl, optionally substituted sulfamoyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C2-C6 heteroalkenyl, optionally substituted C2-C6 heteroalkynyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C4-C 10 Cycloalkenyl, optionally substituted C8-C 10 Cycloalkynyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10 Aryl C1-C6 alkyl, optionally substituted C6-C 10 Aryl C2-C6 alkenyl, optionally substituted C6-C 10 aryl C2-C6 alkynyl, optionally substituted C1-C9 heteroaryl, optionally substituted C1-C9 heteroaryl C1-C6 alkyl, optionally substituted C1-C9 heteroaryl C2-C6 alkenyl, optionally substituted C1-C9 heteroaryl C2-C6 alkynyl, optionally substituted C1-C9 heterocyclyl, optionally substituted C1-C9 heterocyclyl C1-C6 alkyl, optionally substituted C1-C9 heterocyclyl C2-C6 alkenyl, or optionally substituted C1-C9 heterocyclyl C2-C6 alkynyl; X 1 , X 2 , X 3 , X 4 , X 5 , and X 6 Three or fewer of the are N.

[0194] In certain embodiments of formula (IIA), X 3 is CR 4 is.

[0195] In yet another embodiment of Formula (IIA), X 4 is CR 5 is.

[0196] In another embodiment of formula (IIA), X 5 is CR 6 In yet another embodiment of formula (IIA), X 6 is CR 7 is.

[0197] In some embodiments of formula (II) or (IIA), X 1 is N. In certain embodiments of formula (II) or (IIA), X 2 is N.

[0198] In yet another embodiment of Formula (II), the isolated compound has the formula (IIB): TIFF0007625524000011.tif has the structure 38128.

[0199] In some embodiments of formula (IIA) or (IIB), R 4 is H, optionally substituted amino, halo, optionally substituted amido, cyano, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxy, optionally substituted C6-C 10 Aryloxy, optionally substituted C1-C9 heteroaryloxy, optionally substituted C2-C6 alkanoyl, optionally substituted C7-C 11 Aryloyl, optionally substituted C2-C 10 Heteroaryloyl, optionally substituted C2-C 10 Heterocyclyl, hydroxycarbonyl, optionally substituted C2-C7 alkoxycarbonyl, optionally substituted carboxamido, optionally substituted C1-C6 alkanoyloxy, optionally substituted C7-C 11 Aryloyloxy, optionally substituted C2-C 10 Heteroaryloyloxy, optionally substituted C2-C 10Heterocyclyloxy, optionally substituted C1-C6 alkylsulfonyl, optionally substituted C6-C 10 Arylsulfonyl, optionally substituted C1-C9 heteroarylsulfonyl, optionally substituted C1-C9 heterocyclylsulfonyl, optionally substituted sulfamoyl, optionally substituted C1-C6 thioalkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10 In certain embodiments of formula (IIA) or (IIB), R is an aryl C1-C6 alkyl, an optionally substituted C1-C9 heteroaryl, an optionally substituted C1-C9 heteroaryl C1-C6 alkyl, an optionally substituted C1-C9 heterocyclyl, or an optionally substituted C1-C9 heterocyclyl C1-C6 alkyl. 4 is H, optionally substituted amino, halo, optionally substituted amido, cyano, optionally substituted C1-C6 alkyl, optionally substituted C2-C7 alkoxycarbonyl, optionally substituted C2-C7 alkoxycarbonyl, optionally substituted carboxamido, optionally substituted C1-C6 alkanoyloxy, optionally substituted C7-C 11 Aryloyloxy, optionally substituted C2-C 10 Heteroaryloyloxy, optionally substituted C2-C 10 Heterocyclyloxy, optionally substituted C1-C6 alkylsulfonyl, optionally substituted C6-C 10 Arylsulfonyl, optionally substituted sulfamoyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C6-C 10 aryl, optionally substituted C1-C9 heteroaryl, or optionally substituted C1-C9 heterocyclyl. In certain embodiments of formula (IIA) or (IIB), R 4is H, halo, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkylsulfonyl, optionally substituted carboxamido, or optionally substituted sulfamoyl. In other embodiments of Formula (IIA) or (IIB), R 4 is H.

[0200] In still other embodiments of formula (IIA) or (IIB), R 5 is H, optionally substituted amino, halo, optionally substituted amido, optionally substituted carboxamido, cyano, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxy, optionally substituted C6-C 10 Aryloxy, optionally substituted C1-C9 heteroaryloxy, optionally substituted C2-C6 alkanoyl, optionally substituted C7-C 11 Aryloyl, optionally substituted C2-C 10 Heteroaryloyl, optionally substituted C2-C 10 Heterocyclyl, hydroxycarbonyl, optionally substituted C2-C7 alkoxycarbonyl, optionally substituted C1-C6 alkylsulfonyl, optionally substituted C6-C 10 Arylsulfonyl, optionally substituted C1-C9 heteroarylsulfonyl, optionally substituted C1-C9 heterocyclylsulfonyl, optionally substituted C1-C6 thioalkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10 In still other embodiments of Formula (IIA) or (IIB), R is aryl C1-C6 alkyl, optionally substituted C1-C9 heteroaryl, optionally substituted C1-C9 heteroaryl C1-C6 alkyl, optionally substituted C1-C9 heterocyclyl, or optionally substituted C1-C9 heterocyclyl C1-C6 alkyl. 5is H, optionally substituted amino, halo, cyano, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxy, optionally substituted C6-C 10 Aryloxy, optionally substituted C1-C9 heteroaryloxy, optionally substituted C1-C6 heteroalkyl, optionally substituted C6-C 10 In some embodiments of formula (IIA) or (IIB), R is aryl, optionally substituted C1-C9 heteroaryl, or optionally substituted C1-C9 heterocyclyl. 5 is H, optionally substituted amino, halo, optionally substituted C1-C6 alkyl, optionally substituted C6-C 10 aryloxy, optionally substituted C1-C9 heteroaryloxy, or optionally substituted C6-C 10 In certain embodiments of formula (IIA) or (IIB), R 5 is H.

[0201] In certain embodiments of formula (IIA) or (IIB), R 6 is H, optionally substituted amino, halo, optionally substituted amido, optionally substituted carboxamido, cyano, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxy, optionally substituted C6-C 10 Aryloxy, optionally substituted C1-C9 heteroaryloxy, optionally substituted C2-C6 alkanoyl, optionally substituted C7-C 11 Aryloyl, optionally substituted C2-C 10 Heteroaryloyl, optionally substituted C2-C 10 Heterocyclyl, hydroxycarbonyl, optionally substituted C2-C7 alkoxycarbonyl, optionally substituted C1-C6 alkylsulfonyl, optionally substituted C6-C 10Arylsulfonyl, optionally substituted C1-C9 heteroarylsulfonyl, optionally substituted C1-C9 heterocyclylsulfonyl, optionally substituted C1-C6 thioalkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10 In other embodiments of Formula (IIA) or (IIB), R is an aryl C1-C6 alkyl, an optionally substituted C1-C9 heteroaryl, an optionally substituted C1-C9 heteroaryl C1-C6 alkyl, an optionally substituted C1-C9 heterocyclyl, or an optionally substituted C1-C9 heterocyclyl C1-C6 alkyl. 6 is H, optionally substituted amino, halo, optionally substituted amido, optionally substituted carboxamido, optionally substituted C1-C6 alkyl, optionally substituted C2-C7 alkoxycarbonyl, optionally substituted C1-C6 alkoxy, optionally substituted C6-C 10 Aryloxy, optionally substituted C1-C9 heteroaryloxy, optionally substituted C1-C6 heteroalkyl, optionally substituted C6-C 10 In still other embodiments of formula (IIA) or (IIB), R is aryl, optionally substituted C1-C9 heteroaryl, or optionally substituted C1-C9 heterocyclyl. 6 is H, optionally substituted amino, optionally substituted amido, halo, or optionally substituted C1-C6 alkyl. In still other embodiments of Formula (IIA) or (IIB), R 6 is H.

[0202] In some embodiments of formula (IIA) or (IIB), R 7 is H, optionally substituted amino, halo, optionally substituted amido, cyano, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxy, optionally substituted C6-C 10Aryloxy, optionally substituted C1-C9 heteroaryloxy, optionally substituted C2-C6 alkanoyl, optionally substituted C7-C 11 Aryloyl, optionally substituted C2-C 10 Heteroaryloyl, optionally substituted C2-C 10 Heterocyclyl, hydroxycarbonyl, optionally substituted C2-C7 alkoxycarbonyl, optionally substituted carboxamido, optionally substituted C1-C6 alkanoyloxy, optionally substituted C7-C 11 Aryloyloxy, optionally substituted C2-C 10 Heteroaryloyloxy, optionally substituted C2-C 10 Heterocyclyloxy, optionally substituted C1-C6 alkylsulfonyl, optionally substituted C6-C 10 Arylsulfonyl, optionally substituted C1-C9 heteroarylsulfonyl, optionally substituted C1-C9 heterocyclylsulfonyl, optionally substituted sulfamoyl, optionally substituted C1-C6 thioalkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10 In certain embodiments of formula (IIA) or (IIB), R is an aryl C1-C6 alkyl, an optionally substituted C1-C9 heteroaryl, an optionally substituted C1-C9 heteroaryl C1-C6 alkyl, an optionally substituted C1-C9 heterocyclyl, or an optionally substituted C1-C9 heterocyclyl C1-C6 alkyl. 7 is H, optionally substituted amino, halo, optionally substituted amido, cyano, optionally substituted C1-C6 alkyl, optionally substituted C2-C7 alkoxycarbonyl, optionally substituted carboxamido, optionally substituted C1-C6 alkanoyloxy, optionally substituted C7-C 11 Aryloyloxy, optionally substituted C2-C 10Heteroaryloyloxy, optionally substituted C2-C 10 Heterocyclyloxy, optionally substituted C1-C6 alkylsulfonyl, optionally substituted C6-C 10 Arylsulfonyl, optionally substituted sulfamoyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C6-C 10 aryl, optionally substituted C1-C9 heteroaryl, or optionally substituted C1-C9 heterocyclyl. In certain embodiments of formula (IIA) or (IIB), R 7 is H, halo, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkylsulfonyl, optionally substituted carboxamido, or optionally substituted sulfamoyl. In other embodiments of Formula (IIA) or (IIB), R 7 is H.

[0203] In yet another embodiment of Formula (II), the isolated compound has the formula (IIC): It has a structure according to TIFF0007625524000012.tif30128.

[0204] In some embodiments of formula (II), (IIA), (IIB), or (IIC), Ar 1 is an optionally substituted C6 to C 10 In other embodiments of Formula (II), (IIA), (IIB), or (IIC), Ar 1 is an optionally substituted C6 aryl.

[0205] In yet another embodiment of Formula (II), the isolated compound has the formula (IID): It has the structure according to TIFF0007625524000013.tif35128, During the ceremony, R 8 , R 9 , R 10 , R 11 , and R 12each independently represents H, halo, hydroxy, optionally substituted amino, optionally substituted amido, thiol, cyano, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted C1-C6 alkoxy, optionally substituted C6-C 10 Aryloxy, optionally substituted C1-C9 heteroaryloxy, optionally substituted C2-C6 alkanoyl, optionally substituted C7-C 11 Aryloyl, optionally substituted C2-C 10 Heteroaryloyl, optionally substituted C2-C 10 Heterocyclyl, hydroxycarbonyl, optionally substituted C2-C7 alkoxycarbonyl, optionally substituted carboxamido, optionally substituted C1-C6 alkanoyloxy, optionally substituted C7-C 11 Aryloyloxy, optionally substituted C2-C 10 Heteroaryloyloxy, optionally substituted C2-C 10 Heterocyclyloxy, optionally substituted C1-C6 thioalkyl, optionally substituted C1-C6 alkylsulfinyl, optionally substituted C1-C6 alkylsulfonyl, optionally substituted C6-C 10 Arylthio, optionally substituted C6-C 10 Arylsulfinyl, optionally substituted C6-C 10 Arylsulfonyl, optionally substituted C1-C9 heteroarylthio, optionally substituted C1-C9 heteroarylsulfinyl, optionally substituted C1-C9 heteroarylsulfonyl, optionally substituted C1-C9 heterocyclylsulfinyl, optionally substituted C1-C9 heterocyclylsulfonyl, optionally substituted sulfamoyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C2-C6 heteroalkenyl, optionally substituted C2-C6 heteroalkynyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C4-C 10Cycloalkenyl, optionally substituted C8-C 10 Cycloalkynyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10 Aryl C1-C6 alkyl, optionally substituted C6-C 10 Aryl C2-C6 alkenyl, optionally substituted C6-C 10 aryl C2-C6 alkynyl, optionally substituted C1-C9 heteroaryl, optionally substituted C1-C9 heteroaryl C1-C6 alkyl, optionally substituted C1-C9 heteroaryl C2-C6 alkenyl, optionally substituted C1-C9 heteroaryl C2-C6 alkynyl, optionally substituted C1-C9 heterocyclyl, optionally substituted C1-C9 heterocyclyl C1-C6 alkyl, optionally substituted C1-C9 heterocyclyl C2-C6 alkenyl, or optionally substituted C1-C9 heterocyclyl C2-C6 alkynyl; Or, adjacent R 8 , R 9 , R 10 , R 11 , and R 12 Any two of these, taken together with the two adjacent carbon atoms to which they are attached, form a 5-, 6-, or 7-membered optionally substituted carbocyclic or heterocyclic ring.

[0206] In certain embodiments of formula (IID), R 8 is H, halo, or optionally substituted C1-C6 alkyl. In another embodiment of formula (IID), R 8 is H.

[0207] In certain embodiments of formula (IID), R 11 is H, halo, or optionally substituted C1-C6 alkyl. In another embodiment of formula (IID), R 11 is H.

[0208] In some embodiments of Formula (IID), R 12is H, halo, or optionally substituted C1-C6 alkyl. In another embodiment of formula (IID), R 12 is H.

[0209] In certain embodiments of formula (IID), R 9 is H, optionally substituted amino, halo, optionally substituted amido, optionally substituted carboxamido, cyano, nitro, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxy, optionally substituted C6-C 10 Aryloxy, optionally substituted C1-C9 heteroaryloxy, optionally substituted C2-C6 alkanoyl, optionally substituted C7-C 11 Aryloyl, optionally substituted C2-C 10 Heteroaryloyl, optionally substituted C2-C 10 Heterocyclyl, hydroxycarbonyl, optionally substituted C2-C7 alkoxycarbonyl, optionally substituted C1-C6 alkylsulfinyl, optionally substituted C1-C6 alkylsulfonyl, optionally substituted C6-C 10 Arylsulfinyl, optionally substituted C6-C 10 Arylsulfonyl, optionally substituted C1-C9 heteroarylsulfinyl, optionally substituted C1-C9 heteroarylsulfonyl, optionally substituted C1-C9 heterocyclylsulfonyl, optionally substituted sulfamoyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10 In another embodiment of formula (IID), R is an aryl C1-C6 alkyl, an optionally substituted C1-C9 heteroaryl, an optionally substituted C1-C9 heteroaryl C1-C6 alkyl, an optionally substituted C1-C9 heterocyclyl, or an optionally substituted C1-C9 heterocyclyl C1-C6 alkyl. 9is H, optionally substituted amido, halo, cyano, optionally substituted C1-C6 alkyl, optionally substituted C2-C7 alkoxycarbonyl, optionally substituted C1-C6 alkylsulfonyl, optionally substituted C6-C 10 Arylsulfonyl, optionally substituted C1-C9 heteroarylsulfonyl, optionally substituted C1-C9 heterocyclylsulfonyl, optionally substituted sulfamoyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C6-C 10 In yet another embodiment of formula (IID), R is aryl, optionally substituted C1-C9 heteroaryl, or optionally substituted C1-C9 heterocyclyl. 9 is H, optionally substituted carboxamido, halo, optionally substituted C1-C6 alkylsulfonyl, optionally substituted C1-C9 heterocyclylsulfonyl, or optionally substituted sulfamoyl. In still other embodiments of formula (IID), R 9 is an optionally substituted sulfamoyl (eg, unsubstituted sulfamoyl).

[0210] In certain embodiments of formula (IID), R 10 is H, halo, cyano, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted C2-C6 alkanoyl, hydroxycarbonyl, optionally substituted C2-C7 alkoxycarbonyl, optionally substituted carboxamido, optionally substituted C1-C6 thioalkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C6-C 10 In yet another embodiment of formula (IID), R is an aryl C1-C6 alkyl, an optionally substituted C1-C9 heteroaryl C1-C6 alkyl, an optionally substituted C1-C9 heterocyclyl, or an optionally substituted C1-C9 heterocyclyl C1-C6 alkyl. 10is H, halo, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C6-C 10 In yet another embodiment of formula (IID), R is an aryl C1-C6 alkyl, an optionally substituted C1-C9 heteroaryl C1-C6 alkyl, an optionally substituted C1-C9 heterocyclyl, or an optionally substituted C1-C9 heterocyclyl C1-C6 alkyl. 10 is an optionally substituted C1-C6 alkyl, an optionally substituted C1-C6 heteroalkyl, or an optionally substituted C3-C 10 In some embodiments of formula (IID), R 10 may be substituted C 1 ~C 6 It is alkyl (eg, methyl).

[0211] In certain embodiments of formula (II), (IIA), (IIB), (IIC), or (IID), R 1 is H, hydroxy, optionally substituted amino, halo, thiol, optionally substituted amido, optionally substituted carboxamido, cyano, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxy, optionally substituted C6-C 10 Aryloxy, optionally substituted C1-C9 heteroaryloxy, optionally substituted C1-C6 alkanoyloxy, optionally substituted C7-C 11 Aryloyloxy, optionally substituted C2-C 10 Heteroaryloyloxy, optionally substituted C2-C 10 Heterocyclyloxy, optionally substituted C1-C6 thioalkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10In certain embodiments of formula (IID), R is an aryl C1-C6 alkyl, an optionally substituted C1-C9 heteroaryl, an optionally substituted C1-C9 heteroaryl C1-C6 alkyl, an optionally substituted C1-C9 heterocyclyl, or an optionally substituted C1-C9 heterocyclyl C1-C6 alkyl. 10 is H, optionally substituted amino, optionally substituted amido, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxy, optionally substituted C6-C 10 Aryloxy, optionally substituted C1-C9 heteroaryloxy, optionally substituted C1-C6 heteroalkyl, optionally substituted C6-C 10 In some embodiments of Formula (II), (IIA), (IIB), (IIC), or (IID), R is an aryl C1-C6 alkyl, an optionally substituted C1-C9 heteroaryl C1-C6 alkyl, an optionally substituted C1-C9 heterocyclyl, or an optionally substituted C1-C9 heterocyclyl C1-C6 alkyl. 1 is H, optionally substituted amino, optionally substituted C-C 10 Aryloxy, optionally substituted C1-C9 heteroaryloxy, optionally substituted C6-C 10 In other embodiments of formula (II), (IIA), (IIB), (IIC), or (IID), R is aryl, optionally substituted C1-C9 heteroaryl, or optionally substituted C1-C9 heterocyclyl. 1 is optionally substituted amino. In yet another embodiment of formula (IID), R 10 is a substituted amino, wherein at least one of the substituents is phenyl. In still other embodiments of formula (II), (IIA), (IIB), (IIC), or (IID), R 1 is a substituted amino, wherein at least one of the substituents is o-tolyl.

[0212] In some embodiments of Formula (II), the compound of Formula (II) has the formula (IIE): TIFF0007625524000014.tif33128 structure, During the ceremony, R A is an optionally substituted phenyl, and Ar 1 is an optionally substituted phenyl.

[0213] In some embodiments of Formula (IIE), R A is phenyl or 2-methylphenyl.

[0214] In some embodiments of Formula (IIE), Ar 1 is 3-aminosulfonyl-4-methylphenyl.

[0215] In some embodiments of Formula (II), (IIA), (IIB), or (IIC), the isolated compound is compound 19 or 20: The file is TIFF0007625524000015.tif61128.

[0216] In some embodiments, the GPR174 inhibitory compound has formula (III): TIFF0007625524000016.tif21128 or a stereoisomer, or tautomer thereof, or a pharmaceutically acceptable salt thereof; During the ceremony, R 1 and R 2 each is independently H, halo, cyano, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl; and Ar 1 and Ar 2 each independently represents an optionally substituted C6 to C 10 It is aryl or an optionally substituted C1-C9 heteroaryl.

[0217] In certain embodiments of formula (III), Ar 1 is an optionally substituted C6 to C 10In another embodiment of Formula (III), Ar 1 is an optionally substituted C6 aryl, for example, an optionally substituted phenyl.

[0218] In certain embodiments of formula (III), Ar 2 is an optionally substituted C6 to C 10 In another embodiment of Formula (III), Ar 2 is an optionally substituted C6 aryl, for example, an optionally substituted phenyl.

[0219] In yet another embodiment of Formula (III), the isolated compound has the formula (IIIA): It has the structure according to TIFF0007625524000017.tif36128, During the ceremony, R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , and R 12 each independently represents H, halo, hydroxy, optionally substituted amino, optionally substituted amido, thiol, cyano, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted C1-C6 alkoxy, optionally substituted C6-C 10 Aryloxy, optionally substituted C1-C9 heteroaryloxy, optionally substituted C2-C6 alkanoyl, optionally substituted C7-C 11 Aryloyl, optionally substituted C2-C 10 Heteroaryloyl, optionally substituted C2-C 10 Heterocyclyl, hydroxycarbonyl, optionally substituted C2-C7 alkoxycarbonyl, optionally substituted carboxamido, optionally substituted C2-C6 alkanoyloxy, optionally substituted C7-C 11Aryloyloxy, optionally substituted C2-C 10 Heteroaryloyloxy, optionally substituted C2-C 10 Heterocyclyloxy, optionally substituted C2-C6 alkanoylamino, optionally substituted C7-C 11 Aryloylamino, optionally substituted C2-C 10 Heteroaryloylamino, optionally substituted C2-C 10 Heterocyclylamino, optionally substituted C1-C6 thioalkyl, optionally substituted C1-C6 alkylsulfinyl, optionally substituted C1-C6 alkylsulfonyl, optionally substituted C6-C 10 Arylthio, optionally substituted C6-C 10 Arylsulfinyl, optionally substituted C6-C 10 Arylsulfonyl, optionally substituted C1-C9 heteroarylthio, optionally substituted C1-C9 heteroarylsulfinyl, optionally substituted C1-C9 heteroarylsulfonyl, optionally substituted C1-C9 heterocyclylsulfinyl, optionally substituted C1-C9 heterocyclylsulfonyl, optionally substituted sulfamoyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C2-C6 heteroalkenyl, optionally substituted C2-C6 heteroalkynyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C4-C 10 Cycloalkenyl, optionally substituted C8-C 10 Cycloalkynyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10 Aryl C1-C6 alkyl, optionally substituted C6-C 10 Aryl C2-C6 alkenyl, optionally substituted C6-C 10It is aryl C2-C6 alkynyl, optionally substituted C1-C9 heteroaryl, optionally substituted C1-C9 heteroaryl C1-C6 alkyl, optionally substituted C1-C9 heteroaryl C2-C6 alkenyl, optionally substituted C1-C9 heteroaryl C2-C6 alkynyl, optionally substituted C1-C9 heterocyclyl, optionally substituted C1-C9 heterocyclyl C1-C6 alkyl, optionally substituted C1-C9 heterocyclyl C2-C6 alkenyl, or optionally substituted C1-C9 heterocyclyl C2-C6 alkynyl.

[0220] In certain embodiments of formula (III) or (IIIA), R 1 is H, halo, or optionally substituted C1-C6 alkyl. In other embodiments of formula (III) or (IIIA), R 1 is H, halo, or methyl. In still other embodiments of Formula (III) or (IIIA), R 1 is H.

[0221] In certain embodiments of formula (III) or (IIIA), R 2 is H, halo, or optionally substituted C1-C6 alkyl. In other embodiments of formula (III) or (IIIA), R 2 is H, halo, or methyl. In still other embodiments of Formula (III) or (IIIA), R 2 is H.

[0222] In some embodiments of Formula (III) or (IIIA), the isolated compound has the formula (IIIB): It has a structure according to TIFF0007625524000018.tif39128.

[0223] In certain embodiments of formula (III), (IIIA), or (IIIB), R 3 is H, halo, or optionally substituted C1-C6 alkyl. In other embodiments of Formula (III), (IIIA), or (IIIB), R 3 is H.

[0224] In certain embodiments of formula (III), (IIIA), or (IIIB), R 4 is H, halo, or optionally substituted C1-C6 alkyl. In other embodiments of Formula (III), (IIIA), or (IIIB), R 4 is H.

[0225] In some embodiments of formula (III), (IIIA), or (IIIB), R 7 is H, halo, or optionally substituted C1-C6 alkyl. In other embodiments of Formula (III), (IIIA), or (IIIB), R 7 is H.

[0226] In other embodiments of Formula (III), (IIIA), or (IIIB), the isolated compound has the formula (IIIC): It has the structure according to TIFF0007625524000019.tif39128.

[0227] In certain embodiments of formula (III), (IIIA), (IIIB), or (IIIC), R 11 is H, halo, or optionally substituted C1-C6 alkyl. In other embodiments of Formula (III), (IIIA), (IIIB), or (IIIC), R 11 is H.

[0228] In certain embodiments of formula (III), (IIIA), (IIIB), or (IIIC), R 12 is H, halo, or optionally substituted C1-C6 alkyl. In other embodiments of Formula (III), (IIIA), (IIIB), or (IIIC), R 12 is H.

[0229] In some embodiments of Formula (III), (IIIA), (IIIB), or (IIIC), R 8 is H, halo, or optionally substituted C1-C6 alkyl. In other embodiments of Formula (III), (IIIA), (IIIB), or (IIIC), R8 is H.

[0230] In other embodiments of Formula (III), (IIIA), (IIIB), or (IIIC), the isolated compound has the formula (IIID): It has a structure according to TIFF0007625524000020.tif30128.

[0231] In some embodiments of formula (III), (IIIA), (IIIB), (IIIC), or (IIID), R 5 is H, halo, cyano, optionally substituted amino, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkanoyl, optionally substituted C7-C 11 Aryloyl, optionally substituted C2-C 10 Heteroaryloyl, optionally substituted C2-C 10 Heterocyclyl, optionally substituted C2-C6 alkanoyloxy, optionally substituted C7-C 11 Aryloyloxy, optionally substituted C2-C 10 Heteroaryloyloxy, optionally substituted C2-C 10 Heterocyclyloxy, optionally substituted C2-C6 alkanoylamino, optionally substituted C7-C 11 Aryloylamino, optionally substituted C2-C 10 Heteroaryloylamino, optionally substituted C2-C 10 Heterocyclylamino, hydroxycarbonyl, optionally substituted carboxamido, optionally substituted C1-C6 alkylsulfonyl, substituted C6-C 10 Arylsulfonyl, optionally substituted C1-C9 heteroarylsulfonyl, optionally substituted C1-C9 heterocyclylsulfonyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10In other embodiments of Formula (III), (IIIA), (IIIB), (IIIC), or (IIID), R is an aryl C1-C6 alkyl, an optionally substituted C1-C9 heteroaryl, an optionally substituted C1-C9 heteroaryl C1-C6 alkyl, an optionally substituted C1-C9 heterocyclyl, or an optionally substituted C1-C9 heterocyclyl C1-C6 alkyl. 5 is an optionally substituted amino, an optionally substituted C2-C6 alkanoyloxy, an optionally substituted C7-C 11 Aryloyloxy, optionally substituted C2-C 10 Heteroaryloyloxy, optionally substituted C2-C 10 Heterocyclyloxy, optionally substituted C2-C6 alkanoylamino, optionally substituted C7-C 11 Aryloylamino, optionally substituted C2-C 10 Heteroaryloylamino, optionally substituted C2-C 10 In still other embodiments of Formula (III), (IIIA), (IIIB), (IIIC), or (IIID), R is heterocyclylamino, hydroxycarbonyl, or an optionally substituted carboxamido. 5 is an optionally substituted amino, an optionally substituted C2-C6 alkanoylamino, an optionally substituted C7-C 11 Aryloylamino, optionally substituted C2-C 10 Heteroaryloylamino or optionally substituted C2-C 10 It is heterocyclylamino.

[0232] In certain embodiments of formula (III), (IIIA), (IIIB), (IIIC), or (IIID), R 10 is H, halo, cyano, optionally substituted amino, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkanoyl, optionally substituted C7-C 11 Aryloyl, optionally substituted C2-C 10 Heteroaryloyl, optionally substituted C2-C10 Heterocyclyl, optionally substituted C2-C6 alkanoyloxy, optionally substituted C7-C 11 Aryloyloxy, optionally substituted C2-C 10 Heteroaryloyloxy, optionally substituted C2-C 10 Heterocyclyloxy, optionally substituted C2-C6 alkanoylamino, optionally substituted C7-C 11 Aryloylamino, optionally substituted C2-C 10 Heteroaryloylamino, optionally substituted C2-C 10 Heterocyclylamino, hydroxycarbonyl, optionally substituted carboxamido, optionally substituted C1-C6 alkylsulfonyl, substituted C6-C 10 Arylsulfonyl, optionally substituted C1-C9 heteroarylsulfonyl, optionally substituted C1-C9 heterocyclylsulfonyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10 In other embodiments of Formula (III), (IIIA), (IIIB), (IIIC), or (IIID), R is an aryl C1-C6 alkyl, an optionally substituted C1-C9 heteroaryl, an optionally substituted C1-C9 heteroaryl C1-C6 alkyl, an optionally substituted C1-C9 heterocyclyl, or an optionally substituted C1-C9 heterocyclyl C1-C6 alkyl. 10 is an optionally substituted amino, an optionally substituted C2-C6 alkanoyloxy, an optionally substituted C7-C 11 Aryloyloxy, optionally substituted C2-C 10 Heteroaryloyloxy, optionally substituted C2-C 10 Heterocyclyloxy, optionally substituted C2-C6 alkanoylamino, optionally substituted C7-C 11 Aryloylamino, optionally substituted C2-C 10Heteroaryloylamino, optionally substituted C2-C 10 In still other embodiments of Formula (III), (IIIA), (IIIB), (IIIC), or (IIID), R is heterocyclylamino, hydroxycarbonyl, or an optionally substituted carboxamido. 10 is an optionally substituted amino, an optionally substituted C2-C6 alkanoylamino, an optionally substituted C7-C 11 Aryloylamino, optionally substituted C2-C 10 Heteroaryloylamino or optionally substituted C2-C 10 It is heterocyclylamino.

[0233] In certain embodiments of Formula (III), (IIIA), (IIIB), (IIIC), or (IIID), the isolated compound has the formula (IIIE): It has the structure according to TIFF0007625524000021.tif33128, During the ceremony, R A and R B are each independently H or an optionally substituted C1-C6 alkyl, and R C and R D each independently represents H, an optionally substituted C1-C6 alkyl, an optionally substituted C3-C 10 Cycloalkyl, optionally substituted C6-C 10 It is aryl, optionally substituted C1-C9 heteroaryl, or optionally substituted C1-C9 heterocyclyl.

[0234] In some embodiments of formula (III), (IIIA), (IIIB), (IIIC), (IIID), or (IIIE), R A is H. In other embodiments of formula (III), (IIIA), (IIIB), (IIIC), (IIID), or (IIIE), R B is H.

[0235] In certain embodiments of formula (III), (IIIA), (IIIB), (IIIC), (IIID), or (IIIE), R C is an optionally substituted C6 to C 10 aryl, optionally substituted C1-C9 heteroaryl. In other embodiments of formula (III), (IIIA), (IIIB), (IIIC), (IIID), or (IIIE), R C is an optionally substituted C4 heteroaryl, for example, thiophen-2-yl.

[0236] In other embodiments of formula (III), (IIIA), (IIIB), (IIIC), (IIID), or (IIIE), R D is an optionally substituted C6 to C 10 In still other embodiments of formula (III), (IIIA), (IIIB), (IIIC), (IIID), or (IIIE), R D is an optionally substituted C4 heteroaryl, for example, thiophen-2-yl.

[0237] In certain embodiments of formula (III), (IIIA), (IIIB), (IIIC), (IIID), or (IIIE), R 6 is H, halo, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10 In other embodiments of Formula (III), (IIIA), (IIIB), (IIIC), (IIID), or (IIIE), R is an aryl C1-C6 alkyl, an optionally substituted C1-C9 heteroaryl, an optionally substituted C1-C9 heteroaryl C1-C6 alkyl, an optionally substituted C1-C9 heterocyclyl, or an optionally substituted C1-C9 heterocyclyl C1-C6 alkyl. 6is H, halo, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted C1-C6 heteroalkyl, or optionally substituted C3-C 10 In other embodiments of formula (III), (IIIA), (IIIB), (IIIC), (IIID), or (IIIE), R 6 is H, halo, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted C1-C6 heteroalkyl, or optionally substituted C3-C 10 In still other embodiments of formula (III), (IIIA), (IIIB), (IIIC), (IIID), or (IIIE), R 6 is H or an optionally substituted C1-C6 alkyl. In some embodiments of formula (III), (IIIA), (IIIB), (IIIC), or (IIID), R 6 is H. In other embodiments of formula (III), (IIIA), (IIIB), (IIIC), (IIID), or (IIIE), R 6 is C1-C6 alkyl, for example, methyl.

[0238] In certain embodiments of formula (III), (IIIA), (IIIB), (IIIC), (IIID), or (IIIE), R 9 is H, halo, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10In other embodiments of Formula (III), (IIIA), (IIIB), (IIIC), (IIID), or (IIIE), R is an aryl C1-C6 alkyl, an optionally substituted C1-C9 heteroaryl, an optionally substituted C1-C9 heteroaryl C1-C6 alkyl, an optionally substituted C1-C9 heterocyclyl, or an optionally substituted C1-C9 heterocyclyl C1-C6 alkyl. 9 is H, halo, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted C1-C6 heteroalkyl, or optionally substituted C3-C 10 In other embodiments of formula (III), (IIIA), (IIIB), (IIIC), (IIID), or (IIIE), R 9 is H, halo, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted C1-C6 heteroalkyl, or optionally substituted C3-C 10 In still other embodiments of formula (III), (IIIA), (IIIB), (IIIC), (IIID), or (IIIE), R 9 is H or an optionally substituted C1-C6 alkyl. In some embodiments of formula (III), (IIIA), (IIIB), (IIIC), (IIID), or (IIIE), R 9 is H. In other embodiments of formula (III), (IIIA), (IIIB), (IIIC), (IIID), or (IIIE), R 9 is C1-C6 alkyl, for example, methyl.

[0239] In some embodiments, the compound of formula (III) has formula (IIIF): It has the structure TIFF0007625524000022.tif28128, During the ceremony, R C and R Deach independently is an optionally substituted C1-C9 heteroaryl; and R 6 and R 9 Each of these is independently an optionally substituted C1-C6 alkyl.

[0240] In some embodiments of Formula (IIIF), R C and R D each independently represents an unsubstituted C1-C9 heteroaryl; R 6 and R 9 Each of is independently an unsubstituted C1-C6 alkyl.

[0241] In some embodiments of Formula (IIIF), R C and R D Each of is thien-2-yl.

[0242] In some embodiments of Formula (IIIF), R 6 and R 9 Each of is methyl.

[0243] In some embodiments of Formula (III), (IIIA), (IIIB), (IIIC), (IIID), or (IIIE), the isolated compound is Compound 21: The file is TIFF0007625524000023.tif25128.

[0244] In some embodiments, the GPR174 inhibitory compound has formula (IV): TIFF0007625524000024.tif40128 or a stereoisomer thereof, or a tautomer thereof, or a pharmaceutically acceptable salt thereof; During the ceremony, R 1 and R 2each independently represents H, hydroxy, halo, optionally substituted amino, optionally substituted amido, thiol, cyano, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted C1-C6 alkoxy, optionally substituted C6-C 10 Aryloxy, optionally substituted C1-C9 heteroaryloxy, optionally substituted C2-C6 alkanoyl, optionally substituted C7-C 11 Aryloyl, optionally substituted C2-C 10 Heteroaryloyl, optionally substituted C2-C 10 Heterocyclyl, hydroxycarbonyl, optionally substituted C2-C7 alkoxycarbonyl, optionally substituted carboxamido, optionally substituted C1-C6 alkanoyloxy, optionally substituted C7-C 11 Aryloyloxy, optionally substituted C2-C 10 Heteroaryloyloxy, optionally substituted C2-C 10 Heterocyclyloxy, optionally substituted C1-C6 thioalkyl, optionally substituted C1-C6 alkylsulfinyl, optionally substituted C1-C6 alkylsulfonyl, optionally substituted C6-C 10 Arylthio, optionally substituted C6-C 10 Arylsulfinyl, optionally substituted C6-C 10 Arylsulfonyl, optionally substituted C1-C9 heteroarylthio, optionally substituted C1-C9 heteroarylsulfinyl, optionally substituted C1-C9 heteroarylsulfonyl, optionally substituted C1-C9 heterocyclylsulfinyl, optionally substituted C1-C9 heterocyclylsulfonyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C2-C6 heteroalkenyl, optionally substituted C2-C6 heteroalkynyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C4-C 10Cycloalkenyl, optionally substituted C8-C 10 Cycloalkynyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10 Aryl C1-C6 alkyl, optionally substituted C6-C 10 Aryl C2-C6 alkenyl, optionally substituted C6-C 10 aryl C2-C6 alkynyl, optionally substituted C1-C9 heteroaryl, optionally substituted C1-C9 heteroaryl C1-C6 alkyl, optionally substituted C1-C9 heteroaryl C2-C6 alkenyl, optionally substituted C1-C9 heteroaryl C2-C6 alkynyl, optionally substituted C1-C9 heterocyclyl, optionally substituted C1-C9 heterocyclyl C1-C6 alkyl, optionally substituted C1-C9 heterocyclyl C2-C6 alkenyl, or optionally substituted C1-C9 heterocyclyl C2-C6 alkynyl; R 3 and R 4 each independently represents H, hydroxy, halo, optionally substituted amino, optionally substituted amido, thiol, cyano, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted C1-C6 alkoxy, optionally substituted C6-C 10 Aryloxy, optionally substituted C1-C9 heteroaryloxy, optionally substituted C2-C6 alkanoyl, optionally substituted C7-C 11 Aryloyl, optionally substituted C2-C 10 Heteroaryloyl, optionally substituted C2-C 10 Heterocyclyl, hydroxycarbonyl, optionally substituted C2-C7 alkoxycarbonyl, optionally substituted carboxamido, optionally substituted C1-C6 alkanoyloxy, optionally substituted C7-C 11 Aryloyloxy, optionally substituted C2-C 10 Heteroaryloyloxy, optionally substituted C2-C10 Heterocyclyloxy, optionally substituted C1-C6 thioalkyl, optionally substituted C1-C6 alkylsulfonyl, optionally substituted C6-C 10 Arylthio, optionally substituted C6-C 10 Arylsulfonyl, optionally substituted C1-C9 heteroarylthio, optionally substituted C1-C6 heteroalkyl, optionally substituted C2-C6 heteroalkenyl, optionally substituted C2-C6 heteroalkynyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C4-C 10 Cycloalkenyl, optionally substituted C8-C 10 Cycloalkynyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10 Aryl C1-C6 alkyl, optionally substituted C6-C 10 Aryl C2-C6 alkenyl, optionally substituted C6-C 10 aryl C2-C6 alkynyl, optionally substituted C1-C9 heteroaryl, optionally substituted C1-C9 heteroaryl C1-C6 alkyl, optionally substituted C1-C9 heteroaryl C2-C6 alkenyl, optionally substituted C1-C9 heteroaryl C2-C6 alkynyl, optionally substituted C1-C9 heterocyclyl, optionally substituted C1-C9 heterocyclyl C1-C6 alkyl, optionally substituted C1-C9 heterocyclyl C2-C6 alkenyl, or optionally substituted C1-C9 heterocyclyl C2-C6 alkynyl; R 5 is H, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted C1-C6 alkoxy, optionally substituted C6-C 10 Aryloxy, optionally substituted C1-C9 heteroaryloxy, optionally substituted C2-C6 alkanoyl, optionally substituted C7-C 11 Aryloyl, optionally substituted C2-C10 Heteroaryloyl, optionally substituted C2-C 10 Heterocyclyl, optionally substituted C1-C6 alkoxycarbonyl, optionally substituted C1-C6 alkylsulfinyl, optionally substituted C1-C6 alkylsulfonyl, optionally substituted C6-C 10 Arylsulfinyl, optionally substituted C6-C 10 Arylsulfonyl, optionally substituted C1-C9 heteroarylsulfinyl, optionally substituted C1-C9 heteroarylsulfonyl, optionally substituted C1-C9 heterocyclylsulfinyl, optionally substituted C1-C9 heterocyclylsulfonyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C2-C6 heteroalkenyl, optionally substituted C2-C6 heteroalkynyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C4-C 10 Cycloalkenyl, optionally substituted C8-C 10 Cycloalkynyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10 Aryl C1-C6 alkyl, optionally substituted C6-C 10 Aryl C2-C6 alkenyl, optionally substituted C6-C 10 aryl C2-C6 alkynyl, optionally substituted C1-C9 heteroaryl, optionally substituted C1-C9 heteroaryl C1-C6 alkyl, optionally substituted C1-C9 heteroaryl C2-C6 alkenyl, optionally substituted C1-C9 heteroaryl C2-C6 alkynyl, optionally substituted C1-C9 heterocyclyl, optionally substituted C1-C9 heterocyclyl C1-C6 alkyl, optionally substituted C1-C9 heterocyclyl C2-C6 alkenyl, or optionally substituted C1-C9 heterocyclyl C2-C6 alkynyl; n is 0, 1, 2, 3, or 4; and m is 0, 1, 2, 3, 4, 5, or 6.

[0245] In some embodiments of Formula (IV), m is 0.

[0246] In another embodiment of formula (IV), the isolated compound has formula (IVA): It has a structure according to TIFF0007625524000025.tif44128.

[0247] In certain embodiments of formula (IV) or (IVA), R 1 is H, halo, optionally substituted amino, optionally substituted amido, thiol, cyano, or optionally substituted C1-C6 alkyl. In other embodiments of formula (IV) or (IVA), R 1 is H, halo, or optionally substituted C1-C6 alkyl. In still other embodiments of formula (IV) or (IVA), R 1 is H.

[0248] In certain embodiments of Formula (IV) or Formula (IVA), the isolated compound has the formula (IVB): It has the structure according to TIFF0007625524000026.tif43128.

[0249] In some embodiments of formula (IV), (IVA), or (IVB), R 5 is H, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted C2-C6 alkanoyl, optionally substituted C7-C 11 Aryloyl, optionally substituted C2-C 10 Heteroaryloyl, optionally substituted C2-C 10 Heterocyclyl, optionally substituted C1-C6 alkyloxycarbonyl, optionally substituted C1-C6 alkylsulfonyl, or optionally substituted C6-C 10 In other embodiments of formula (IV), (IVA), or (IVB), R 5is H, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkanoyl, optionally substituted C7-C 11 In still other embodiments of Formula (IV), (IVA), or (IVB), R is aryloyl, or optionally substituted C1-C6 alkyloxycarbonyl. 5 is H, optionally substituted C1-C6 alkyl, or optionally substituted C2-C6 alkanoyl. In still other embodiments of Formula (IV), (IVA), or (IVB), R 5 is H.

[0250] In certain embodiments of Formula (IV), (IVA), or (IVB), the isolated compound has the formula (IVC): It has the structure according to TIFF0007625524000027.tif43128.

[0251] In certain embodiments of formula (IV), (IVA), (IVB), or (IVC), R 2 is H, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C2-C6 heteroalkenyl, optionally substituted C2-C6 heteroalkynyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C4-C 10 Cycloalkenyl, optionally substituted C8-C 10 Cycloalkynyl, optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10 Aryl C1-C6 alkyl, optionally substituted C6-C 10 Aryl C2-C6 alkenyl, optionally substituted C6-C 10In other embodiments of Formula (IV), (IVA), (IVB), or (IVC), R is an aryl C2-C6 alkynyl, optionally substituted C1-C9 heteroaryl, optionally substituted C1-C9 heteroaryl C1-C6 alkyl, optionally substituted C1-C9 heteroaryl C2-C6 alkenyl, optionally substituted C1-C9 heteroaryl C2-C6 alkynyl, optionally substituted C1-C9 heterocyclyl, optionally substituted C1-C9 heterocyclyl C1-C6 alkyl, optionally substituted C1-C9 heterocyclyl C2-C6 alkenyl, or optionally substituted C1-C9 heterocyclyl C2-C6 alkynyl. 2 is an optionally substituted C3 to C 10 Cycloalkyl, optionally substituted C4-C 10 Cycloalkenyl, optionally substituted C8-C 10 Cycloalkynyl, optionally substituted C6-C 10 In still other embodiments of formula (IV), (IVA), (IVB), or (IVC), R is aryl, optionally substituted C1-C9 heteroaryl, or optionally substituted C1-C9 heterocyclyl. 2 is an optionally substituted C6 to C 10 In still other embodiments of formula (IV), (IVA), (IVB), or (IVC), R is aryl, optionally substituted C1-C9 heteroaryl, or optionally substituted C1-C9 heterocyclyl. 2 is an optionally substituted C6 to C 10 In other embodiments of formula (IV), (IVA), (IVB), or (IVC), R 2 is an optionally substituted pyridyl (e.g., 2-pyridyl, 3-pyridyl, or 4-pyridyl). In certain embodiments of formula (IV), (IVA), (IVB), or (IVC), R 2 is an optionally substituted phenyl.

[0252] In some embodiments of Formula (IV), (IVA), (IVB), or (IVC), the isolated compound has the formula (IVD): It has the structure according to TIFF0007625524000028.tif51128, During the ceremony, R 6 is independently in each occurrence halo, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C6-C 10 Aryl, optionally substituted C2-C6 heteroaryl, optionally substituted C2-C6 heterocyclyl, optionally substituted C2-C6 alkynyl, optionally substituted amino, optionally substituted amido, thiol, cyano, nitro, C1-C6 alkylsulfonyl, hydroxycarbonyl, optionally substituted C2-C7 alkoxycarbonyl, optionally substituted C6-C 10 aryloxy, or optionally substituted C2-C6 heteroaryloxy; Z 1 is C or N; Z 2 is C or N; Z 3 is N or C; and p is 0, 1, 2, 3, 4, or 5.

[0253] In some embodiments of Formula (IVD), Z 1 is C and Z 2 is C and Z 3 is N. In another embodiment of formula (IVD), Z 1 is C and Z 2 is N and Z 3 is C. In certain embodiments of formula (IVD), Z 1 is N and Z 2 is C and Z 3 is C. In certain other embodiments of formula (IVD), Z 1 is C and Z 2 is C and Z 3 is C.

[0254] In some embodiments of Formula (IV), (IVA), (IVB), or (IVC), the isolated compound has the formula (IVD): It has the structure according to TIFF0007625524000029.tif51128, During the ceremony, R 6 is independently in each occurrence halo, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C6-C 10 Aryl, optionally substituted C2-C6 heteroaryl, optionally substituted C2-C6 heterocyclyl, optionally substituted C2-C6 alkynyl, optionally substituted amino, optionally substituted amido, thiol, cyano, nitro, C1-C6 alkylsulfonyl, hydroxycarbonyl, optionally substituted C2-C7 alkoxycarbonyl, optionally substituted C2-C6 alkoxy, optionally substituted C2-C6 alkenoxy, optionally substituted C6-C 10 aryloxy, or optionally substituted C2-C6 heteroaryloxy; Z 1 is CH or N; Z 2 is CH or N; Z 3 is N or CH; and p is 0, 1, 2, 3, 4, or 5.

[0255] In some embodiments of Formula (IVD), Z 1 is C and Z 2 is C and Z 3 is N. In another embodiment of formula (IVD), Z 1 is C and Z 2 is N and Z 3 is CH. In certain embodiments of formula (IVD), Z 1 is N and Z 2 is CH and Z 3 is CH. In certain other embodiments of formula (IVD), Z 1 is CH and Z 2 is CH and Z 3is CH.

[0256] In some embodiments of Formula (IVD), p is 0. In other embodiments of Formula (IVD), p is 1. In particular embodiments of Formula (IVD), p is 2. In some embodiments of Formula (IVD), p is 1 and R 6 is present in the p or m position.

[0257] In certain embodiments of formula (IVD), R 6 is methoxy, methyl, hydroxyl, ethoxy, ethyl, optionally substituted phenoxy, optionally substituted cyclopentyloxy, t-butoxy, allyloxy, isopropyloxy, n-pentyloxy, trifluoromethyloxy, difluoromethyloxy, fluoro, chloro, nitro, 2-hydroxyethyloxy, optionally substituted 1,3,4-oxadiazolyl, or optionally substituted pyrrolidyl.

[0258] In certain embodiments of formula (IV), (IVA), (IVB), (IVC), or (IVD), R 3 is H, halo, optionally substituted amino, optionally substituted amido, thiol, cyano, hydroxycarbonyl, optionally substituted C2-C7 alkoxycarbonyl, or optionally substituted C1-C6 alkyl.

[0259] In other embodiments of formula (IV), (IVA), (IVB), (IVC), or (IVD), n is 0.

[0260] In some embodiments of Formula (IV), the compound has the formula (IVE): TIFF0007625524000030.tif45128 structure, During the ceremony, Z 2 and Z 3 Each of these is independently 6 or N; and R 6each independently represents H, halogen, hydroxy, nitro, optionally substituted C1-C6 alkylsulfonyl, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxy, optionally substituted C4-C 11 Cycloalkoxy, optionally substituted C1-C6 haloalkoxy, optionally substituted C2-C6 alkenoxy, optionally substituted C6-C 10 Aryloxy, optionally substituted C1-C9 heterocyclyl, optionally substituted C6-C 10 aryl or optionally substituted C1-C9 heteroaryl; Or, two adjacent R 6 The groups together with the carbon atoms to which they are attached form a C1-C9 heterocyclyl.

[0261] In some embodiments of Formula IV, (IVA), (IVB), or (IVC), the isolated compound is compound 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 54, or 55: TIFF0007625524000031.tif211119TIFF0007625524000032.tif243119TIFF0007625524000033.tif239119TIFF0007625524000034.tif188128TIFF0007625524000035.tif119128.

[0262] In some embodiments, the compound of formula (IVE) is compound 54 or 55: The file is TIFF0007625524000036.tif53128.

[0263] In some embodiments, the GPR174 inhibitory compound has the following formula (V): TIFF0007625524000037.tif24128 or a stereoisomer thereof, or a tautomer thereof, or a pharmaceutically acceptable salt thereof; During the ceremony, R 1 is phenyl, and R 2 is an optionally substituted C6 to C 10 It is aryl or an optionally substituted C1-C9 heteroaryl.

[0264] In some embodiments of Formula (V), R 2 is an optionally substituted phenyl.

[0265] In some embodiments of Formula (V), R 2 is phenyl substituted with para-C2-C6 alkenoxy.

[0266] In some embodiments of Formula (V), R 2 is phenyl substituted with para-(2-methylallyl)oxy.

[0267] In some embodiments of Formula (V), the compound of Formula (V) is compound 56: TIFF0007625524000038.tif30128.

[0268] In some embodiments, the GPR174 inhibitory compound has the following formula (Va): TIFF0007625524000039.tif23128, or a stereoisomer thereof, or a tautomer thereof, or a pharmaceutically acceptable salt thereof; During the ceremony, X is O or S; R 1a is optionally substituted phenyl; and R 2a is an optionally substituted C6 to C 10 aryl, optionally substituted C3-C9 heteroaryl, or optionally substituted C3-C 10It is heteroarylalkyl.

[0269] In some embodiments, R 1a is a substituted phenyl. In some more specific embodiments, R 1a is optionally substituted with halo (e.g., F, Br, Cl, or I). In some embodiments, R 1a has the following structure: I have TIFF0007625524000040.tif16128.

[0270] In some embodiments, R 2a is an optionally substituted C3 to C 10 In some embodiments, R is heteroarylalkyl. 2a is unsubstituted. In some more particular embodiments, R 2a has the following structure: I have TIFF0007625524000041.tif12128.

[0271] In some embodiments, X is O. In certain embodiments, X is S.

[0272] In some embodiments, the compound of formula (VI): Provided herein is a GPR174 inhibitory compound described in TIFF0007625524000042.tif26128 or a stereoisomer thereof, or a tautomer thereof, or a pharmaceutically acceptable salt thereof; During the ceremony, R 1 is an optionally substituted C1-C9 heteroaryl, and R 2 is a halogen.

[0273] In some embodiments of formula (VI), the N=C bond has the (E) configuration.

[0274] In some embodiments of formula (VI), the N=C bond has the (Z) configuration.

[0275] In some embodiments of Formula (VI), R 1 is optionally substituted pyridinyl or optionally substituted furanyl.

[0276] In some embodiments of Formula (VI), R 1 is pyridin-4-yl.

[0277] In some embodiments of Formula (VI), R 1 is 2,5-dimethyl-fur-3-yl.

[0278] In some embodiments of Formula (VI), R 2 is a halo.

[0279] In some embodiments of Formula (VI), R 2 is chloro or bromo.

[0280] In some embodiments of Formula (VI), the compound of Formula (VI) is compound 57 or 58: TIFF0007625524000043.tif31134 or a stereoisomer thereof, or a tautomer thereof, or a pharmaceutically acceptable salt thereof.

[0281] In any of the embodiments described herein, the compound can be a compound described in Table 1 (eg, any of compounds 1-59).

[0282] In another aspect, the present disclosure provides a method for identifying a modulatory agent capable of modulating the GPR174-mediated Gs signaling pathway, comprising: (a) contacting a cell with a candidate modulatory agent, wherein the cell expresses GPR174 and comprises a GPR174-mediated signaling pathway; and (b) determining whether the candidate modulatory agent modulates the GPR174-mediated Gs signaling pathway relative to cells contacted with a control or relative to a reference standard; In some embodiments, the candidate modulator is a chemical compound comprising a structure according to any of Formulas I-VI disclosed herein. In certain embodiments, step (b) further comprises performing an assay to detect at least one of transcriptional reporter gene expression, GTPase activity, cAMP levels, intracellular messenger levels, calcium levels, downstream kinase activity, transcriptional activation of downstream genes, changes in cell morphology, changes in cell proliferation rate, arachidonic acid release, or extracellular acidification rate.

[0283] In some embodiments, the candidate modulator is selected from the group consisting of a chemical compound, a nucleic acid, a natural extract, and a gas. In certain embodiments, the compound is a small molecule compound. In certain embodiments, the chemical compound comprises a structure according to any of Formulas I-VI disclosed herein.

[0284] In another aspect, the present disclosure provides a method for identifying a modulatory agent capable of modulating GPR174-mediated Gs signaling pathway activity, comprising: (a) Cells expressing GPR174, (i) at least one candidate modulator; and (ii) reference chemical compounds known to modulate GPR174-mediated Gs signaling pathway activity; contacting the (b) measuring the level of GPR174-mediated signaling pathway activity in the cells contacted by step (a); and (c) comparing the level of GPR174-mediated signaling pathway activity in cells contacted with only the reference chemical compound with the GPR174-mediated signaling pathway activity measured in step (b), wherein a difference in GPR174-mediated signaling pathway activity between cells containing the candidate modulator in the presence of the reference compound and cells contacted with only the reference compound indicates that the candidate modulator is capable of modulating GPR174 activity. The present invention is characterized by a method comprising:

[0285] In some embodiments, the reference chemical compound known to modulate GPR174-mediated Gs signaling pathway activity comprises a structure according to any of Formulas I-VI disclosed herein. In certain embodiments, step (b) further comprises performing an assay to detect at least one of transcriptional reporter gene expression, GTPase activity, cAMP levels, intracellular messenger levels, calcium levels, downstream kinase activity, transcriptional activation of downstream genes, changes in cell morphology, changes in cell proliferation rate, arachidonic acid release, or extracellular acidification rate. In some embodiments, the candidate modulator is selected from the group consisting of chemical compounds, nucleic acids, natural extracts, and gases. In certain embodiments, the compound is a chemical compound. In certain embodiments, the small molecule compound comprises a structure according to any of Formulas I-VI.

[0286] In another aspect, the disclosure features a method for identifying a modulator capable of modulating GPR174-mediated signaling pathway activity. The method includes the steps of: (a) contacting cells expressing GPR174 with (i) at least one candidate modulator and (ii) a reference chemical compound known to modulate GPR174-mediated signaling pathway activity; (b) measuring the level of GPR174-mediated signaling pathway activity in the cells contacted in step (a); and (c) comparing the level of GPR174-mediated signaling pathway activity in the cells contacted with only the reference chemical compound with the GPR174-mediated signaling pathway activity measured in step (b), wherein a difference in GPR174-mediated signaling activity between cells containing the candidate modulator in the presence of the reference compound and cells contacted with only the reference compound indicates that the candidate modulator can modulate GPR174 activity. In some embodiments, the reference chemical compound comprises a structure according to any one of Formulas I-VI (e.g., any of Compounds 1-59).

[0287] In certain embodiments, step (b) further comprises performing an assay to detect at least one of transcriptional reporter gene expression, GTPase activity, cAMP levels, intracellular messenger levels, calcium levels, downstream kinase activity, transcriptional activation of downstream genes, changes in cell morphology, changes in cell proliferation rate, arachidonic acid release, or extracellular acidification rate.

[0288] In some embodiments, the candidate modulator is selected from the group consisting of a chemical compound, a nucleic acid, a natural extract, and a gas. In certain embodiments, the compound is a small molecule chemical compound.

[0289] In another aspect, the disclosure features a method of using a small molecule chemical compound to inhibit a GPR174-mediated signaling pathway in a cell. The method includes: (a) providing a small molecule chemical compound, wherein the small molecule chemical compound functionally interacts with GPR174 and inhibits the GPR174-mediated Gs signaling pathway in cells expressing GPR174, and the compound meets the following criteria: (i) the compound has a structure selected from the group consisting of Formulas I, II, III, IV, V, and VI (or (I), (II), (III), (IV), (V), (Va), or (VI)); or (ii) the compound alters the binding affinity of any one of reference compounds 1-59 (listed in Table 1) to GPR174; or (iii) the compound, when tested in the presence of the reference compound, differentially inhibits the inhibitory activity of any one of reference compounds 1-59 (listed in Table 1) in a GPR174-mediated signaling pathway assay compared to the inhibitory activity of the reference compound alone: characterized by at least one of the steps of: (b) contacting a cell expressing GPR174, which contains a GPR174-mediated signaling pathway, with the compound described in step (a), thereby inhibiting the GPR174-mediated signaling pathway in the cell. Includes:

[0290] In another aspect, the disclosure features a method of using a small molecule chemical compound to inhibit a GPR174-mediated signaling pathway in a cell. The method includes: (a) providing a small molecule chemical compound, wherein the small molecule chemical compound functionally interacts with GPR174 and inhibits the GPR174-mediated Gs signaling pathway in cells expressing GPR174, and the compound meets the following criteria: (i) the compound has a structure selected from the group consisting of Formulas I, II, III, IV, V, and VI (or (I), (II), (III), (IV), (V), (Va), or (VI)); or (ii) the compound alters the apparent binding affinity of any one of reference compounds 1-59 (listed in Table 1) to GPR174; or (iii) said compound, when tested in the presence of said reference compound, differentially inhibits the inhibitory activity of any one of reference compounds 1-59 (listed in Table 1) in a GPR174-mediated signaling pathway assay compared to the inhibitory activity of the reference compound alone: characterized by at least one of the steps of: (b) contacting a cell expressing GPR174, which contains a GPR174-mediated signaling pathway, with the compound described in step (a), thereby inhibiting the GPR174-mediated signaling pathway in the cell. Includes:

[0291] In some embodiments, the compound alters the binding affinity of any one of reference compounds 1-59 (listed in Table 1) to GPR174. In certain embodiments, the compound decreases (i.e., inhibits) the binding affinity of any one of reference compounds 1-59 to GPR174. In certain embodiments, the compound competitively decreases (i.e., competitively inhibits) the binding affinity of any one of reference compounds 1-59 to GPR174. In other embodiments, the compound increases the binding affinity of any one of reference compounds 1-59 (listed in Table 1) to GPR174.

[0292] In some embodiments, the compound alters the apparent binding affinity of any one of reference compounds 1-59 (listed in Table 1) to GPR174. In certain embodiments, the compound decreases the apparent binding affinity of any one of reference compounds 1-59 to GPR174. In certain embodiments, the compound competitively binds to GPR174, decreasing the apparent binding affinity of any one of reference compounds 1-59 to GPR174. In alternative embodiments, the compound allosterically binds to GPR174, decreasing the apparent binding affinity of any one of reference compounds 1-59 to GPR174. In other embodiments, the compound increases the binding affinity of any one of reference compounds 1-59 (listed in Table 1) to GPR174.

[0293] In certain embodiments of any of the above aspects of the disclosure, the GPR174-mediated signaling pathway is the Gs pathway.

[0294] In some embodiments of any of the above aspects, the cell is present in a mammalian subject in need of treatment for any of the conditions, diseases, or disorders described herein.

[0295] Inhibition of ATP-adenosine A2aR and / or A2bR mediated signaling (such as A2aR antagonists, and / or A2bR antagonists and / or CD73 inhibitors and / or CD38 inhibitors and / or CD39 inhibitors) Adenosine signaling, particularly via the A2A and / or A2B receptor pathways, has been identified as a promising approach for cancer immunotherapy. See Leone et al., “Targeting adenosine for cancer immunotherapy,” J. ImmunoTher. Cancer., 2018, 6:57. Allard et al., “The ectonucleotidases CD39 and CD73: novel checkpoint inhibitor targets,” Immunol. Rev., 2017, 276(1), 121-144 (see also Table 15 below). Adenosine is an immunosuppressive metabolite produced at high levels within the tumor microenvironment. Adenosine signaling has emerged as a key metabolic pathway regulating tumor immunity. In particular, adenosine signaling via the A2A receptor expressed on immune cells potently attenuates immune responses in inflamed tissues. Adenosine is produced under hypoxic conditions in inflamed and malignant tissues through the conversion of ATP mediated by the enzymes CD73 (NT5E, 5'-nucleotidase (5'-NT) or ecto-5'-nucleotidase) and CD39 (ectonucleoside triphosphate diphosphohydrolase-1, NTPDase1). Adenosine can also be produced from NAD+ by a CD38-centered axis of NAD+ metabolism that generates adenosine diphosphate ribose (ADPR). Adenosine generated through the CD38-mediated pathway has been shown to correlate with human myeloma progression (Horenstein et al., Mol Med 22:694-704, 2016). Adenosine production and signaling, particularly via A2A receptors, plays a role in the resolution of inflammation in response to tissue injury and dampens the immune response. However, this coupling of wound healing and immunosuppression represents a mechanism of cancer immune evasion. Therefore, inhibition of the hypoxia-CD38-CD39-CD73-A2AR pathway has been identified as a promising target for cancer immunotherapy.

[0296] Inhibitors of the ATP-adenosine A2A and / or A2B receptor pathway Inhibitors of the ATP-adenosine A2A receptor pathway and / or inhibitors of the ATP-adenosine A2B receptor pathway that can be used in combination with GPR174 inhibitors in the compositions, methods, and uses described herein include adenosine A2A receptor antagonists, A2B receptor antagonists, and inhibitors of enzymes that degrade ATP to adenosine, such as inhibitors of CD73 and inhibitors of CD38 and inhibitors of CD39, and combinations of such agents.

[0297] Adenosine A2A receptor antagonist Adenosine A2A receptor antagonists include the compounds ATL-444, AZD4635, caffeine, ciforadenant (CPI-444; V81444), CPI-445, EOS100850, MK-3814, istradefylline (KW-6002), MSX-3, PBF-509 (NIR178), preladenant (SCH-420,814), SCH-58261, SCH-412,348, SCH-442,416, ST-1535, ST-4206, theophylline, tozadenant (SYN115), VER-6623, VER-6947, VER-7835, bipadenant (BIIB-014), and ZM-241,385, whose structures are shown below. TIFF0007625524000044.tif214154TIFF0007625524000045.tif142151

[0298] Additional A2A receptor antagonists are described in Preti, et al., "History and Perspectives of A2A Adenosine Receptor Antagonists as Potential Therapeutic Agents," Med. Res. Rev., 2015, 35(4), 790-848, and Congreve et al., "Targeting adenosine A2A receptor antagonism for treatment of cancer," Exp. Opin. Drug Discov., 2018, 13(11), 997-1003, the disclosures of which are incorporated herein by reference.

[0299] Other A2A receptor antagonists are described in the following international patent applications, the disclosures of which are incorporated herein by reference: PCT Publication Nos. WO 2019 / 007140; WO 2018 / 166493; WO 2018 / 161910; WO 2018 / 130184; WO 2018 / 059531; WO 2017 / 136375; WO 2017 / 112917; WO 2017 / 008205; WO 2017 / 011214; WO 2016 / 200717; WO 2016 / 126570; WO 2016 / 0 87429; WO 2016 / 081290; WO 2015 / 020565; WO 2014 / 105664; WO 2014 / 105666; WO 2014 / 101120; WO 2014 / 101113; WO 2013 / 156614, WO 2013 / 058681; WO 2012 / 129381; WO 2012 / 112962; WO 2012 / 061787; WO WO 2012 / 060844; WO 2012 / 038980; WO 2011 / 061527; WO 2011 / 060207; WO 2011 / 053507; WO 2010 / 040003; WO 2010 / 037122; WO 2009 / 055308; WO 2009 / 050198; WO 2008 / 055711; WO 2007 / 047293; WO 2007 / 038212; WO 2006 / 137527; WO 2006 / 129626; WO 2006 / 124770; WO 2006 / 083949; WO 2012 / 03898; WO 2011 / 06152; WO 2011 / 06020; WO 2011 / 05350; WO 2010 / 04000; WO 2010 / 03712; WO 2009 / 055308; WO 2009 / 050198;WO 2008 / 055711; WO 2007 / 047293; WO 2007 / 038212; WO 2006 / 137527; WO 2006 / 129626; WO 2006 / 124770; and WO 2006 / 083949. ;

[0300] Adenosine A2B receptor antagonist Adenosine A2B receptor antagonists include the compounds MRS-1754, GS-6201, ISAM-140, PSB-0788, PSB-1115, and PSB-603, the structures of which are as follows: Shown in TIFF0007625524000046.tif118137.

[0301] Additional A2B receptor antagonists include ATL-801, CVT-6883, MRS-1706, OSIP-339,391, PSB-1901, PBF-1129, and Vigano S. et al., Frontiers in Immunology vol 10:925,2019; Volpini R. et al., Journal of Med Chem 45(15):3271-9,2002; Volpini R. et al., Current Pharmaceutical Design 8(25):2285-98,2002; Baraldi PG et al., Journal of Med Chem 47(6):1434-47,2004; Cacciari B. et al., Mini Reviews in Med Chem 5(12):1053-60,2005; Baraldi PG et al., Current Med Chem 13(28):3467-82, 2006; Beukers MW et al., Medicinal Research Reviews, 26(5):667-98, 2006; Elzein E. et al., Bioorganic & Medicinal Chemistry Letters 16(2):302-6, 2006; Carotti A. et al., Journal of Med Chem 49(1):282-99, 2006; Tabrizi MA et al., Bioorganic & Medicinal Chemistry 16(5):2419-30, 2008; Stefanachi A. et al., Bioorganic & Medicinal Chemistry 16(6):2852-69, 2008 and Jiang et al., Journal of Med Chem 62(8):4032-4055, 2019, as well as further A2B receptor antagonists described in Stefanachi A compound 38 described in A. et al., Bioorganic & Medicinal Chemistry 16(22):9780-9, 2008 is included.

[0302] Other A2B receptor antagonists are described in the following international patent applications, the disclosures of which are incorporated herein by reference: PCT Publication No. WO 1995 / 011681; WO 1999 / 042093; WO 2000 / 049051; WO 2000 / 073307; WO 2009 / 157938; WO 2011 / 005871; WO 2012 / 112964; WO 2007 / 149277; WO 2019 / 123482; WO 2007 / 134958; WO 2005 / 051951; WO 2003 / 042214; WO 2007 / 039297; WO 2005 / 040155; WO 2005 / 042534; WO 2004 / 106337; WO 2001 / 016134; WO 2008 / 027585; WO 2003 / 053366; WO 2003 / 053361; WO 2005 / 070926; WO 2008 / 080461; WO 2016 / 150901; WO 2005 / 051951; WO 2003 / 042214; WO 2003 / 063800; and WO 2016 / 164838.

[0303] CD73 (NT5E, 5'-nucleotidase (5'-NT) or ecto-5'-nucleotidase) inhibitors CD73 inhibitors can include CD73 antibodies, nucleotides that inhibit CD73 expression (eg, inhibitory RNA), as well as chemical (eg, small molecule) inhibitors.

[0304] CD73 antibodies, including oleculab, are described in the following international patent applications, the disclosures of which are incorporated herein by reference: PCT Publication Nos. WO 2018 / 237157; WO 2018 / 237173; WO 2018 / 215535; WO 2018 / 187484; WO 2018 / 137598; WO 2018 / 013611; WO 2017 / 152085; WO 2017 / 118613; WO 2017 / 100670; WO 2017 / 064043; WO 2016 / 131950; WO 2016 / 081748; WO 2016 / 075176; WO 2016 / 075099; and WO 2016 / 055609.

[0305] Nucleotides that inhibit CD73 expression are described in the following international patent application, the disclosure of which is incorporated herein by reference: PCT Publication No. WO 2018 / 065627.

[0306] CD73 inhibitors include N-benzyl-α,β-methylene adenosine 5'-diphosphate sodium salt and α,β-methylene adenosine 5'-diphosphate sodium salt (PSB 12379), BMS-986179, MEDI9447, CPI-006, and NZV 930. CD73 inhibitors are also described in the following international patent applications, the disclosures of which are incorporated herein by reference: PCT Publication No. WO 2018 / 208727; WO 2018 / 208980; WO 2018 / 187512; WO 2018 / 183635; WO 2018 / 110555; WO 2018 / 094148; WO 2018 / 067424; WO 2017 / 153952; WO 2017 / 120508; WO 2017 / 098421; WO 2015 / 164573; WO 2015 / 049447; and WO 2007 / 135195.

[0307] It is noted that while CD73 inhibitors are useful in the compositions and methods described herein for the treatment of cancer, the use of CD73 inhibitors is preferably avoided during early tumor formation and in the setting of metastasis due to the fact that adenosine derived from CD73 may help prevent metastasis (it may act as a barrier in the vascular endothelium), and therefore, in such situations it may be preferable to leave CD73 in an active state.

[0308] CD38 (cyclic ADP-ribose hydrolase) inhibitor CD38 is a surface enzyme that catalyzes the synthesis and hydrolysis of cyclic ADP-ribose (cADPR) from NAD+ to ADP-ribose, in addition to the synthesis of NAADP from NADP+.

[0309] CD38 inhibitors can include CD38 antibodies, nucleotides that inhibit CD38 expression (eg, inhibitory RNA), as well as chemical (eg, small molecule) inhibitors.

[0310] CD38 antibodies, including daratumumab, and other CD38 inhibitors are described in the following publications and international patent applications, the disclosures of which are incorporated herein by reference: Xia C et al., Drugs of Today. 52(10):551-560, 2016; Escande C. et al., Diabetes, 62(4):1084-1093, 2013; U.S. Patent No. 9,840,496; U.S. Patent Application Publication No. 2018 / 85383; U.S. Patent Application Publication No. 2017 / 0260164; PCT Publication No. WO 2019 / 140410; WO 20190 / 74973; WO 2019 / 0904931; WO 2019 / 087151; WO 2019 / 074973; WO 2019 / 034753; WO 2019 / 034752; WO 2019 / 020643; WO 2018 / 224685; WO 2018 / 224683; WO 2018 / 224682; WO 2016 / 087975; and WO 2013 / 002879.

[0311] CD39 (ectonucleoside triphosphate diphosphohydrolase-1, NTPDase1) inhibitor CD39 inhibitors can include CD39 antibodies, nucleotides that inhibit CD39 expression (eg, inhibitory RNA), as well as chemical (eg, small molecule) inhibitors.

[0312] CD39 antibodies, including TTX-030, are described in the following international patent applications, the disclosures of which are incorporated herein by reference: PCT Publication Nos. WO 2018 / 167267; WO 2018 / 065552; ​​WO 2017 / 191300; WO 2017 / 157948; WO 2017 / 089334; WO 2017 / 064043; WO 2016 / 073845; WO 2012 / 085132; and WO 2009 / 095478.

[0313] Nucleotides that inhibit CD39 expression are described in the following international patent application, the disclosure of which is incorporated herein by reference: PCT Publication No. WO 2018 / 065622.

[0314] CD39 inhibitors include sodium metatungstate (POM1); 6-N,N-diethyl-D-β,γ-dibromomethylene ATP trisodium salt (ARL-67156); 1-amino-4-(1-naphthyl)aminoanthraquinone-2-sulfonic acid sodium salt (PSB 06126), 1-amino-4-(4-chlorophenyl)aminoanthraquinone-2-sulfonic acid sodium salt (PSB069), and IPH52. CD39 inhibitors are also described in the following international patent application, the disclosure of which is incorporated herein by reference: PCT Publication No. WO 2007 / 135195.

[0315] Other features and advantages of the present disclosure will become apparent from the following detailed description, drawings, and claims. [Brief explanation of the drawings]

[0316] [Figure 1A] The amino acid sequence of human GPR174 (NP_115942.1) is set forth as SEQ ID NO: 1. [Figure 1B] 1 shows the cDNA (NM_032553.1) encoding human GPR174, which is set forth as SEQ ID NO:2. [Figure 1C] The amino acid sequence of mouse GPR174 (NP_001028423.1) is set forth as SEQ ID NO: 3. [Figure 1D] The amino acid sequence of rat GPR174 (NP_001100408.1) is set forth as SEQ ID NO: 4. [Figure 2] 1 is a graph showing that the reported potency ranking of various known agonists for the muscarinic acetylcholine receptor M1 (CHRM1) receptor is conserved in a cell redistribution assay (CRA). [Figure 3] FIG. 1 is a schematic diagram showing the algorithm used to determine the G protein signaling profile of orphan GPCRs. [Figure 4A] 1 is a graph showing the effect of various compounds on beta-2 adrenergic receptor (ADBR2) signaling. [Figure 4B] 1 is a graph showing the effect of ADBR2 overexpression on the CRE-luc reporter, as measured by transient transfection of increasing amounts (ng) of DNA encoding ADBR2. [Figure 4C] 1 is a graph showing the effect of ADBR2 overexpression on the NFAT-luc reporter, as measured by transient transfection of increasing amounts (ng) of DNA encoding ADBR2. [Figure 4D] 1 is a graph demonstrating that the compound ICI 118551, an inverse agonist of ADBR2, specifically inhibits CRE-luc reporter activity in cells transiently transfected with ADBR2. [Figure 4E] 1 is a graph demonstrating that the compound ICI 118551 does not inhibit NFAT-luc reporter activity in cells transiently transfected with ADBR2. [Figure 5A] 1 is a graph illustrating dose-response curves of representative compounds, including compounds 1, 2, 3, 4, and 20, against GPR174 in a Cellular Redistribution Assay (CRA) assay. [Figure 5B] 1 is a graph illustrating dose-response curves for representative compounds, including compounds 1, 2, 3, 4, and 20, to CHRM1 in a CRA assay. The dose-response curve for the CHRM1 interacting compound pirenzepine is also shown. [Figure 6] 6A-6B are graphs showing that Compound 1 (Group I) inhibited the Gs pathway in the presence of GPR174 (FIG. 6A), but not in the presence of ADBR2 (FIG. 6B). [Figure 7]7A-7B are graphs showing that compound 2 (Group I) inhibited the Gs pathway in the presence of GPR174 (FIG. 7A), but not in the presence of ADBR2 (FIG. 7B). [Figure 8] 8A-8B are graphs showing that compound 4 (Group I) did not regulate the Gs pathway in the presence of GPR174 (FIG. 8A) and did not regulate the Gs pathway in the presence of ADBR2 (FIG. 8B). [Figure 9] 9A-9B are graphs showing that compound 6 (Group I) inhibited the Gs pathway in the presence of GPR174 (FIG. 9A), but not in the presence of ADBR2 (FIG. 9B). [Figure 10] 10A-10B are graphs showing that compound 7 (Group I) inhibited the Gs pathway in the presence of GPR174 (FIG. 10A), but not in the presence of ADBR2 (FIG. 10B). [Figure 11] 11A-11B are graphs showing that compound 10 (Group I) inhibited the Gs pathway in the presence of GPR174 (FIG. 11A), but not in the presence of ADBR2 (FIG. 11B). [Figure 12] 12A-12B are graphs showing that compound 11 (Group I) inhibited the Gs pathway in the presence of GPR174 (FIG. 12A), but not in the presence of ADBR2 (FIG. 12B). [Figure 13] 13A-13B are graphs showing that compound 19 (Group II) did not regulate the Gs pathway in the presence of GPR174 (FIG. 13A) and did not regulate the Gs pathway in the presence of ADBR2 (FIG. 13B). [Figure 14] 14A-14B are graphs showing that compound 21 (Group III) did not regulate the Gs pathway in the presence of GPR174 (FIG. 14A) and did not regulate the Gs pathway in the presence of ADBR2 (FIG. 14B). [Figure 15] 15A-15B are graphs showing that compound 22 (Group IV) inhibited the Gs pathway in the presence of GPR174 (FIG. 15A), but not in the presence of ADBR2 (FIG. 15B). [Figure 16] 16A-16B are graphs showing that compound 23 (Group IV) inhibited the Gs pathway in the presence of GPR174 (FIG. 16A), but not in the presence of ADBR2 (FIG. 16B). [Figure 17] 17A-17B are graphs showing that compound 31 (Group IV) inhibited the Gs pathway in the presence of GPR174 (FIG. 17A), but not in the presence of ADBR2 (FIG. 17B). [Figure 18] 18A-18B are graphs showing that compound 33 (Group IV) inhibited the Gs pathway in the presence of GPR174 (FIG. 18A), but not in the presence of ADBR2 (FIG. 18B). [Figure 19] 19A-19B are graphs showing that compound 36 (Group IV) inhibited the Gs pathway in the presence of GPR174 (FIG. 19A), but not in the presence of ADBR2 (FIG. 19B). [Figure 20] 20A-20B are graphs showing that compound 42 (Group IV) inhibited the Gs pathway in the presence of GPR174 (FIG. 20A), but not in the presence of ADBR2 (FIG. 20B). [Figure 21] Figures 21A-21B are graphs showing that compound 4 (Group I) does not regulate the Gs pathway in the presence of GPR174 but competes with the GPR174 agonist LysoPS (Figure 21A), and that neither compound 4 nor LysoPS regulates Gs signaling in the presence of ADBR (Figure 21B). [Figure 22] 1 graphically illustrates the relative transcript abundance of GPR174 in human tissues as measured by qPCR, as described in Example 5. [Figure 23] 1 graphically illustrates the relative transcript abundance of GPR174 in human lymphoid cells as measured by qPCR, as described in Example 5. [Figure 24]Figure 24A graphically depicts the percentage of FoxP3+Helios- cells in the CD4+ cell population in human peripheral blood mononuclear cell (PBMC) cultures treated with vehicle, Compound 10, or interleukin-2 (IL-2) on day 3 post-stimulation (p=0.03 for Compound 10 vs. vehicle; p=0.003 for IL-2 vs. vehicle), as described in Example 6. Figure 24B graphically depicts the percentage of FoxP3+Helios+ cells in the CD4+ cell population on day 7 post-stimulation in PBMC cultures treated with vehicle, Compound 10, or IL-2, as described in Example 6. (p=0.01 for Compound 10 vs. vehicle; p=0.01 for IL-2 vs. vehicle). [Figure 25] Graph shows the amount of IL-2 (fold increase over vehicle levels) in culture supernatants on day 2 after stimulation in PBMC cultures treated with vehicle or Compound 10 (1 μM, 3 μM, or 10 μM), as described in Example 6. [Figure 26] Graph shows the amount of IFN-γ (fold increase over vehicle levels) in culture supernatants on day 2 post-stimulation in PBMC cultures treated with vehicle or Compound 10 (1 μM, 3 μM, or 10 μM), as described in Example 6. [Figure 27] 12A-12D are graphs showing the amount of IL-2 (pg / mL) in culture supernatants 48 hours after stimulation in splenocyte cultures from WT or GPR174 KO mice treated with vehicle or Compound 10 (10 μM), as described in Example 7. [Figure 28] The graph shows the percentage of FoxP3+Helios+ cells in the CD4+ cell population at day 7 post-stimulation in human PBMC cultures from a single donor treated with vehicle or Compound 10 (3 μM, 10 μM, or 30 μM) as described in Example 8 (n=3, *p<0.05; **p<0.01). [Figure 29]13A-13C are graphs showing the amount of IL-2 (pg / mL) in the culture supernatant of human PBMC cultures from a single donor treated with vehicle or GPR174 inhibitory compound 10 (3 μM, 10 μM, 30 μM, or 60 μM) on day 3 post-stimulation, as described in Example 8. [Figure 30] 12A-C graphically show the percentage of vehicle FoxP3+Helios+ cells in the CD4+ cell population at day 7 post-stimulation in human PBMC cultures from eight different donors treated with vehicle or Compound 10 (30 μM), as described in Example 9. [Figure 31] 12A-C graphically show the amount of IL-2 (fold over IL-2 produced by vehicle) in culture supernatants at 2 days post-stimulation in human PBMC cultures from eight different donors treated with vehicle or Compound 10 (30 μM), as described in Example 9. [Figure 32] Graph showing the amounts of various cytokines (fold over vehicle-produced cytokines) in culture supernatants from human PBMCs obtained from eight donors treated with vehicle or Compound 10 (30 μM) on day 2 post-stimulation, as described in Example 9. Statistically significant fold increases were observed for interleukin-6 (IL-6) (***p<0.001), interleukin-10 (IL-10) (*p<0.05), interferon gamma (IFN-γ) (*p<0.05), and tumor necrosis factor (TNF-α) (*p<0.05) compared to vehicle controls, and a statistically significant fold decrease was observed for interleukin-17 (IL-17A) (***p<0.001) compared to vehicle controls. [Figure 33] This graph shows the fold induction by Compound 10 (30 μM) of a panel of cytokines in co-cultured murine C57BL / 6+DBA1 splenocytes on day 6 after mixing, compared to the cytokine levels measured on day 6 in C57BL / 6+DBA1 splenocytes co-cultured in the absence of Compound 10, as described in Example 10. [Figure 34A]1 shows the CD4+ (46.2%), CD8+ (17.1%), and non-T cell (33.9%) populations present in a representative PBMC culture from a single donor 4 hours after stimulation with anti-CD3 and anti-CD28 antibodies in the presence of Compound 10 (10 μM), as described in Example 11. [Figure 34B] FIG. 34B graphically depicts the percentage of cells with intracellular IL-2 in the CD4+ T cell population treated with compound 10 (10 μM) shown in FIG. 34A compared to vehicle control, as described in Example 11. [Figure 34C] 34A , which graphically depicts the percentage of cells with intracellular IL-2 in the CD8+ T cell population treated with compound 10 (10 μM) shown in FIG. 34A compared to vehicle control, as described in Example 11. [Figure 35-1] Figure 35A shows the CD4+ (31.8%), CD8+ (27.5%), and non-T cell (33.1%) populations present in a representative PBMC culture from a single donor 2 days after stimulation with anti-CD3 and anti-CD28 antibodies, as described in Example 11. [Figure 35-2] Figure 35B graphically depicts the percentage of cells with intracellular IL-2 staining in the CD4+ T cell population in PBMC cultures treated with compound 10 (0, 0.1, 0.3, 1.0, or 3.0 μM), representatives of which are shown in Figure 35A, as described in Example 11. Figure 35C graphically depicts the percentage of cells with intracellular IL-2 staining in the CD8+ T cell population in PBMC cultures treated with compound 10 (0, 0.1, 0.3, 1.0, or 3.0 μM), representatives of which are shown in Figure 35A, as described in Example 11. Figure 35D graphically depicts the percentage of cells with intracellular IL-2 staining in the non-T cell population in PBMC cultures treated with compound 10 (0, 0.1, 0.3, 1.0, or 3.0 μM), representatives of which are shown in Figure 35A, as described in Example 11. [Figure 35-3]Figure 35E graphically depicts the percentage of cells with intracellular IL-10 staining in the CD4+ T cell population in PBMC cultures treated with compound 10 (0, 0.1, 0.3, 1.0, or 3.0 μM), representatives of which are shown in Figure 35A, as described in Example 11. Figure 35F graphically depicts the percentage of cells with intracellular IL-10 staining in the CD8+ T cell population in PBMC cultures treated with compound 10 (0, 0.1, 0.3, 1.0, or 3.0 μM), representatives of which are shown in Figure 35A, as described in Example 11. Figure 35G graphically depicts the percentage of cells with intracellular IL-10 staining in the non-T cell population in PBMC cultures treated with compound 10 (0, 0.1, 0.3, 1.0, or 3.0 μM), representatives of which are shown in Figure 35A, as described in Example 11. [Figure 35-4] Figure 35H graphically depicts the percentage of cells with intracellular IFN-γ staining in the CD4+ T cell population in PBMC cultures treated with compound 10 (0, 0.1, 0.3, 1.0, or 3.0 μM), representatives of which are shown in Figure 35A, as described in Example 11. Figure 35I graphically depicts the percentage of cells with intracellular IFN-γ staining in the CD8+ T cell population in PBMC cultures treated with compound 10 (0, 0.1, 0.3, 1.0, or 3.0 μM), representatives of which are shown in Figure 35A, as described in Example 11. Figure 35J graphically depicts the percentage of cells with intracellular IFN-γ staining in the non-T cell population in PBMC cultures treated with compound 10 (0, 0.1, 0.3, 1.0, or 3.0 μM), representatives of which are shown in Figure 35A, as described in Example 11. [Figure 35-5]Figure 35K graphically depicts the percentage of cells with intracellular TNF-α staining in the CD4+ T cell population in PBMC cultures treated with compound 10 (0, 0.1, 0.3, 1.0, 3.0, or 10 μM), representatives of which are shown in Figure 35A, as described in Example 11. Figure 35L graphically depicts the percentage of cells with intracellular TNF-α staining in the CD8+ T cell population in PBMC cultures treated with compound 10 (0, 0.1, 0.3, 1.0, 3.0, or 10 μM), representatives of which are shown in Figure 35A, as described in Example 11. Figure 35M graphically depicts the percentage of cells with intracellular TNF-α staining in the non-T cell population in PBMC cultures treated with compound 10 (0, 0.1, 0.3, 1.0, 3.0, or 10 μM), representatives of which are shown in Figure 35A, as described in Example 11. [Figure 36-1] Figure 36A shows further analysis of the non-T cell population from Figure 35A sorted into CD56+CD16- (2.27%), CD56+CD16+ (11.8%), and non-NK (78.5%) populations present in the non-T cell population in a representative PBMC culture 2 days after stimulation with anti-CD3 and anti-CD28 antibodies, as described in Example 11. [Figure 36-2] Figure 36B graphically depicts the percentage of cells with intracellular IFN-γ staining in the non-T-CD56+CD16+ cell population in PBMC cultures treated with compound 10 (0, 0.1, 0.3, 1.0, or 3.0 μM), representatives of which are shown in Figure 36A, as described in Example 11. Figure 36C graphically depicts the percentage of cells with intracellular IFN-γ staining in the non-T-CD56+CD16− cell population in PBMC cultures treated with compound 10 (0, 0.1, 0.3, 1.0, or 3.0 μM), representatives of which are shown in Figure 36A, as described in Example 11. [Figure 36-3]Figure 36D graphically depicts the percentage of cells with intracellular TNF-α staining in the non-T-CD56+CD16+ cell population in PBMC cultures treated with compound 10 (0, 0.1, 0.3, 1.0, 3.0, or 10 μM), representatives of which are shown in Figure 36A, as described in Example 11. Figure 36E graphically depicts the percentage of cells with intracellular TNF-α staining in the non-T-CD56+CD16− cell population in PBMC cultures treated with compound 10 (0, 0.1, 0.3, 1.0, 3.0, or 10 μM), representatives of which are shown in Figure 36A, as described in Example 11. [Figure 37] Graph showing the amount of dextramerstained cells (antigen-specific T cells) as a percentage of total CD8+ cells in splenocyte cultures obtained from WT and GPR174 KO mice 3 weeks after immunization with 1000 HAU influenza virus A / PR / 8 / 34 and cultured for 5 days in the presence of NP antigenic peptide, as described in Example 12; splenocytes obtained from naive WT mice are included as a control. [Figure 38] Graph showing the amount of dextramerstained cells (antigen-specific T cells) as a percentage of total CD8+ cells in splenocyte cultures obtained from WT and GPR174 KO mice immunized with 1000 HAU (hemagglutinin units) of influenza virus A / PR / 8 / 34 and boosted 3 weeks later, as described in Example 12. Splenocytes were analyzed 11 days after the boost; splenocytes obtained from naive WT mice are included as a control. [Figure 39]Figure 39A graphically depicts the proportion of naive regulatory T cells (Tregs) with CTLA-4 positive staining (percentage of naive Treg cells (CD45RA+FOXP3+)) in PBMC cultures treated with GPR174 inhibitory compound 10 (0, 0.19, 0.38, 0.75, 1.5, 3.0, or 6.0 μM) or an equivalent amount of vehicle control (DMSO) one day after stimulation, as described in Example 13. Figure 39B graphically depicts the proportion of non-regulatory T cells (non-Treg) with CTLA4 positive staining (percentage of naive non-Treg cells (CD45RA+FOXP3-)) in PBMC cultures treated with GPR174 inhibitory compound 10 (0, 0.19, 0.38, 0.75, 1.5, 3.0, or 6.0 μM) or an equivalent amount of vehicle control (DMSO) one day after stimulation, as described in Example 13. [Figure 40] Figure 40A graphically depicts the percentage of memory Tregs with CTLA4-positive staining (percentage of memory Tregs (CD45RA-FOXP3+)) in PBMC cultures treated with GPR174 inhibitory compound 10 (0, 0.19, 0.38, 0.75, 1.5, 3.0, or 6.0 μM) or an equivalent amount of vehicle control (DMSO) one day after stimulation, as described in Example 13. Figure 40B graphically depicts the percentage of memory non-Treg cells with CTLA4-positive staining (percentage of memory non-Tregs (CD45RA-FOXP3-)) in PBMC cultures treated with GPR174 inhibitory compound 10 (0, 0.19, 0.38, 0.75, 1.5, 3.0, or 6.0 μM) or an equivalent amount of vehicle control (DMSO) one day after stimulation, as described in Example 13. [Figure 41] 1 is a graph showing the percentage of human CD4+ T cells with positive PD-L1 staining in PBMC cultures treated with GPR174 inhibitor Compound 10 (0, 1.25, or 5 μM) one day after stimulation, as described in Example 14, demonstrating a dose-dependent decrease in PD-L1 expression in CD4+ T cells treated with Compound 10 compared to vehicle control-treated cells. [Figure 42]10 is a graph showing the percentage of human CD8+ T cells with TGIT-positive staining in PBMC cultures treated with GPR174 inhibitory compound 10 (0, 1.25, or 5 μM) one day after stimulation, as described in Example 14, demonstrating a dose-dependent decrease in TGIT expression in CD8+ T cells treated with compound 10 compared to vehicle control-treated cells. [Figure 43] Graph showing the percentage of human CD4+ T cells with AREG-positive staining in PBMC cultures treated with GPR174-inhibitory compound 10 (0, 1.25, or 5 μM) one day after stimulation, as described in Example 14, demonstrating a dose-dependent decrease in AREG expression in CD4+ T cells treated with compound 10 compared to vehicle control-treated cells. [Figure 44] 10 is a graph showing the amount of IFN-γ in the culture supernatant 24 hours after stimulation in human PBMC cultures from a single donor cultured in the presence of GPR174 inhibitory compound 10 (3 μM); A2aR inhibitory compound (ZM241385, 1 μM or 10 μM); a combination of compound 10 (3 μM) + 1 μM ZM241385; a combination of compound 10 (3 μM) + 10 μM ZM241385; or vehicle control (DMSO), as described in Example 15, demonstrating that combined inhibition of GPR174 and A2aR results in synergistic induction of IFN-γ production. [Figure 45] Figure 45A graphically depicts the fold induction of Gs signaling activity in HEK293 cells expressing wild-type GPR174 and the cAMP biosensor expression plasmid pGlo22F in the presence of increasing concentrations of phosphatidylserine liposomes (PS) or lysophosphatidylserine (Lyso-PS), as described in Example 16. Figure 45B graphically depicts the fold induction of Gs signaling activity in HEK293 cells expressing mutant GPR174-v38 and the cAMP biosensor expression plasmid pGlo22F in the presence of increasing concentrations of phosphatidylserine liposomes (PS) or lysophosphatidylserine (Lyso-PS), as described in Example 16. [Figure 46] 10 is a graph showing the level of GPR174 Gs signaling activity (shown as the ratio of luminescence after addition of phospholipids to the pre-read luminescence value) in HEK293 cells expressing GPR174 and the cAMP biosensor expression plasmid pGlo22F in the presence of PS, phosphatidylcholine (PC), phosphatidylethanolamine (PE), and phosphatidylinositol (PI) liposomes, as described in Example 16. [Figure 47A] As described in Example 16, this graph shows that GPR174 inhibitory compound 10 (Group I) inhibits PS-induced GPR174 Gs signaling in a dose-responsive manner with an apparent IC50 of approximately 20-40 nM. [Figure 47B] 10A-10C graphically show that GPR174 inhibitory compound 6 (Group I) inhibits PS-induced GPR174 Gs signaling in a dose-responsive manner with an apparent IC50 of approximately 0.1 μM, as described in Example 16. [Figure 47C] As described in Example 16, this graph shows that GPR174 inhibitory compound 11 (Group I) inhibits PS-induced GPR174 Gs signaling in a dose-responsive manner with an apparent IC50 of approximately 0.1 μM. [Figure 47D] 10A-10C graphically show that GPR174 inhibitory compound 20 (Group II) inhibits PS-induced GPR174 Gs signaling in a dose-responsive manner with an apparent IC50 of approximately 3.0 μM, as described in Example 16. [Figure 47E] As described in Example 16, this graph shows that GPR174 inhibitory compound 30 (Group IV) inhibits PS-induced GPR174 Gs signaling in a dose-responsive manner with an apparent IC50 of approximately 1.3 μM. [Figure 47F]10A-10C graphically show that GPR174 inhibitory compound 23 (Group IV) inhibits PS-induced GPR174 Gs signaling in a dose-responsive manner with an apparent IC50 of approximately 0.15 μM, as described in Example 16. [Figure 48A] Graph shows the ratio of post-addition luminescence to pre-read luminescence of HEK293 cells expressing GPR174 and GloSensor exposed to culture medium or untreated (non-apoptotic) K562 cells or apoptotic K562 cells (treated with HO for 20 hours), as described in Example 17. [Figure 48B] Graph shows the ratio of post-addition luminescence to pre-read luminescence of HEK293 cells expressing GPR174 and GloSensor exposed to culture medium or freshly isolated neutrophils or apoptotic neutrophils (treated with anti-Fas antibody), as described in Example 17. [Figure 48C] Graph showing the ratio of post-addition luminescence to pre-read luminescence of HEK293 cells expressing GPR174 and GloSensor exposed to culture supernatant (Sup) or platelets (Pl) as described in Example 17. [Figure 49A] 10 is a graph showing the amount of IFN-γ (pg / mL) in the culture supernatant 24 hours after stimulation in human PBMC cultures from a single donor (donor 1) cultured in the presence of GPR174 inhibitory compound 10 (2 μM); A2aR inhibitory compounds (SCH-58261, 0.2 μM, 0.6 μM, or 2 μM); a combination of compound 10 (2 μM) + 0.2 μM SCH-58261; a combination of compound 10 (2 μM) + 0.6 μM SCH-58261 or a combination of compound 10 (2 μM) + 2 μM SCH-58261; or a vehicle control (DMSO), as described in Example 18. [Figure 49B]13A-13C are graphs showing the amount of IL-2 (pg / mL) in the culture supernatant 24 hours after stimulation in human PBMC cultures from a single donor (donor 1) cultured in the presence of GPR174 inhibitory compound 10 (2 μM); A2aR inhibitory compounds (SCH-58261, 0.2 μM, 0.6 μM, or 2 μM); a combination of compound 10 (2 μM) + 0.2 μM SCH-58261; a combination of compound 10 (2 μM) + 0.6 μM SCH-58261 or a combination of compound 10 (2 μM) + 2 μM SCH-58261; or a vehicle control (DMSO), as described in Example 18. [Figure 49C] 13 is a graph showing the amount of TNF (pg / mL) in the culture supernatant 24 hours after stimulation in human PBMC cultures from a single donor (donor 1) cultured in the presence of GPR174 inhibitory compound 10 (2 μM); A2aR inhibitory compounds (SCH-58261, 0.2 μM, 0.6 μM, or 2 μM); a combination of compound 10 (2 μM) + 0.2 μM SCH-58261; a combination of compound 10 (2 μM) + 0.6 μM SCH-58261 or a combination of compound 10 (2 μM) + 2 μM SCH-58261; or a vehicle control (DMSO), as described in Example 18. [Figure 50A] 10 is a graph showing the amount of IFN-γ (pg / mL) in the culture supernatant 24 hours after stimulation in human PBMC cultures from a single donor (donor 2) cultured in the presence of GPR174 inhibitory compound 10 (1 μM); A2aR inhibitory compound (ZM, 0.2 μM); A2aR inhibitory compound (PBF-509 0.1 μM); the combination of compound 10 (1 μM) + 0.2 μM ZM; the combination of compound 10 (1 μM) + 0.1 μM PBF-509; or vehicle control (DMSO), as described in Example 18. [Figure 50B]10 is a graph showing the amount of IL-2 (pg / mL) in the culture supernatant 24 hours after stimulation in human PBMC cultures from a single donor (donor 2) cultured in the presence of GPR174 inhibitory compound 10 (1 μM); A2aR inhibitory compound (ZM, 0.2 μM); A2aR inhibitory compound (PBF-509 0.1 μM); the combination of compound 10 (1 μM) + 0.2 μM ZM; the combination of compound 10 (1 μM) + 0.1 μM PBF-509; or vehicle control (DMSO), as described in Example 18. [Figure 50C] This graph shows the amount of TNF (pg / mL) in the culture supernatant 24 hours after stimulation in human PBMC cultures from a single donor (donor 2) cultured in the presence of GPR174 inhibitory compound 10 (1 μM); A2aR inhibitory compound (ZM, 0.2 μM); A2aR inhibitory compound (PBF-509 0.1 μM); the combination of compound 10 (1 μM) + 0.2 μM ZM; the combination of compound 10 (1 μM) + 0.1 μM PBF-509; or vehicle control (DMSO), as described in Example 18. [Figure 51A] 10 is a graph showing the amount of IFN-γ (pg / mL) in the culture supernatant 24 hours after stimulation in human PBMC cultures from a single donor (donor 3) cultured in the presence of GPR174 inhibitory compound 10 (1 μM); A2aR inhibitory compound (ZM, 0.2 μM); A2aR inhibitory compound (PBF-509 0.2 μM); the combination of compound 10 (1 μM) + 0.2 μM ZM; the combination of compound 10 (1 μM) + 0.2 μM PBF-509; or vehicle control (DMSO), as described in Example 18. [Figure 51B]10 is a graph showing the amount of IL-2 (pg / mL) in the culture supernatant 24 hours after stimulation in human PBMC cultures from a single donor (donor 3) cultured in the presence of GPR174 inhibitory compound 10 (1 μM); A2aR inhibitory compound (ZM, 0.2 μM); A2aR inhibitory compound (PBF-509 0.2 μM); the combination of compound 10 (1 μM) + 0.2 μM ZM; the combination of compound 10 (1 μM) + 0.2 μM PBF-509; or vehicle control (DMSO), as described in Example 18. [Figure 51C] Graph showing the amount of TNF (pg / mL) in the culture supernatant 24 hours after stimulation in human PBMC cultures from a single donor (donor 3) cultured in the presence of GPR174 inhibitory compound 10 (1 μM); A2aR inhibitory compound (ZM, 0.2 μM); A2aR inhibitory compound (PBF-509 0.2 μM); the combination of compound 10 (1 μM) + 0.2 μM ZM; the combination of compound 10 (1 μM) + 0.2 μM PBF-509; or vehicle control (DMSO), as described in Example 18. [Figure 52A] Graph showing the fold change from vehicle in the amount of IFN-γ in the culture supernatant 24 hours after stimulation in human PBMC cultures from 12 donors cultured in the presence of GPR174 inhibitory compound 10 (1-2 μM), A2aR inhibitory compound (ZM, 0.2-1 μM); the combination of compound 10 (1-2 μM) + 0.2-1 μM ZM; or vehicle control (DMSO), as described in Example 19. [Figure 52B] 10 is a graph showing the fold change from vehicle in the amount of IL-2 in the culture supernatant 24 hours after stimulation in human PBMC cultures from 12 donors cultured in the presence of GPR174 inhibitory compound 10 (1-2 μM), A2aR inhibitory compound (ZM, 0.2-1 μM); the combination of compound 10 (1-2 μM) + 0.2-1 μM ZM; or vehicle control (DMSO), as described in Example 19. [Figure 52C]Graph showing the fold change from vehicle in the amount of TNF in the culture supernatant 24 hours after stimulation in human PBMC cultures from 12 donors cultured in the presence of GPR174 inhibitory compound 10 (1-2 μM), A2aR inhibitory compound (ZM, 0.2-1 μM); the combination of compound 10 (1-2 μM) + 0.2-1 μM ZM; or vehicle control (DMSO), as described in Example 19. [Figure 52D] Graph showing the fold change from vehicle in the amount of GM-CSF in the culture supernatant 24 hours after stimulation in human PBMC cultures from 12 donors cultured in the presence of GPR174 inhibitory compound 10 (1-2 μM), A2aR inhibitory compound (ZM, 0.2-1 μM); the combination of compound 10 (1-2 μM) + 0.2-1 μM ZM; or vehicle control (DMSO), as described in Example 19. [Figure 53A] 10 is a graph showing the amount of IFN-γ (pg / mL) in the culture supernatant 24 hours after stimulation of human PBMC cultures from a single donor (donor 3) cultured in the presence of GPR174 inhibitory compound 10 (1 μM); ZM, 0.1 μM; PBF (0.1 μM), SCH (0.2 μM), MRS (1 μM), adenosine deaminase (ADA, 0.75 μg / mL), or vehicle control, alone or in combination with compound 10, as described in Example 20. [Figure 53B] 10 is a graph showing the amount of IL-2 (pg / mL) in the culture supernatant 24 hours after stimulation of human PBMC cultures from a single donor (donor 3) cultured in the presence of GPR174 inhibitory compound 10 (1 μM); ZM, 0.1 μM; PBF (0.1 μM), SCH (0.2 μM), MRS (1 μM), adenosine deaminase (ADA, 0.75 μg / mL), or vehicle control, alone or in combination with compound 10, as described in Example 20. [Figure 53C]10 is a graph showing the amount of TNF (pg / mL) in the culture supernatant 24 hours after stimulation of human PBMC cultures from a single donor (donor 3) cultured in the presence of GPR174 inhibitory compound 10 (1 μM); ZM, 0.1 μM; PBF (0.1 μM), SCH (0.2 μM), MRS (1 μM), adenosine deaminase (ADA, 0.75 μg / mL), or vehicle control, alone or in combination with compound 10, as described in Example 20. [Figure 54A] 10 is a graph showing the amount of IFN-γ (pg / mL) in the culture supernatant 24 hours after stimulation of human PBMC cultures from a single donor (donor 6) cultured in the presence of GPR174 inhibitory compound 10 (1 μM); ZM, 0.1 μM; PBF (0.1 μM), SCH (0.2 μM), MRS (1 μM), adenosine deaminase (ADA, 0.75 μg / mL), or vehicle control, alone or in combination with compound 10, as described in Example 20. [Figure 54B] 10 is a graph showing the amount of IL-2 (pg / mL) in the culture supernatant 24 hours after stimulation of human PBMC cultures from a single donor (donor 6) cultured in the presence of GPR174 inhibitory compound 10 (1 μM); ZM, 0.1 μM; PBF (0.1 μM), SCH (0.2 μM), MRS (1 μM), adenosine deaminase (ADA, 0.75 μg / mL), or vehicle control, alone or in combination with compound 10, as described in Example 20. [Figure 54C] 10 is a graph showing the amount of TNF (pg / mL) in the culture supernatant 24 hours after stimulation of human PBMC cultures from a single donor (donor 6) cultured in the presence of GPR174 inhibitory compound 10 (1 μM); ZM, 0.1 μM; PBF (0.1 μM), SCH (0.2 μM), MRS (1 μM), adenosine deaminase (ADA, 0.75 μg / mL), or vehicle control, alone or in combination with compound 10, as described in Example 20. [Figure 55A]2 is a graph showing the amount of IFN-γ (pg / mL) in the supernatant 24 hours after stimulation of human PBMCs (donor 7) in the presence of GPR174 inhibitory compounds 6, 10, 11, or 20, respectively, in the presence or absence of ZM, as described in Example 21. [Figure 55B] 10 graphically depicts the amount of IL-2 (pg / mL) in the supernatant 24 hours after stimulation of human PBMCs (donor 7) in the presence of GPR174 inhibitory compounds 6, 10, 11, or 20, respectively, in the presence or absence of ZM, as described in Example 21. [Figure 55C] 2 is a graph showing the amount of TNF (pg / mL) in the supernatant 24 hours after stimulation of human PBMCs (donor 7) in the presence of GPR174 inhibitory compounds 6, 10, 11, or 20, respectively, in the presence or absence of ZM, as described in Example 21. [Figure 55D] Graph showing the amount of GM-CSF (pg / mL) in the supernatant 24 hours after stimulation of human PBMCs (donor 7) in the presence of GPR174 inhibitory compounds 6, 10, 11, or 20, respectively, in the presence or absence of ZM, as described in Example 21. [Figure 56A] The graph shows the fold change from vehicle in the amount of IFN-γ in the culture supernatant 24 hours after stimulation in human PBMC cultures from five donors cultured in the presence of GPR174 inhibitory compound 10 (1 μM); A2aR inhibitory compound (ZM, 0.2 μM); A2bR inhibitory compound MRS (2 μM); or a combination of compound 10 (1 μM) + 0.2 μM ZM + 2 μM MRS; or vehicle control (DMSO), as described in Example 22. [Figure 56B]The graph shows the fold change from vehicle in the amount of IL-2 in the culture supernatant 24 hours after stimulation in human PBMC cultures from five donors cultured in the presence of GPR174 inhibitory compound 10 (1 μM); A2aR inhibitory compound (ZM, 0.2 μM); A2bR inhibitory compound MRS (2 μM); or a combination of compound 10 (1 μM) + 0.2 μM ZM + 2 μM MRS; or vehicle control (DMSO), as described in Example 22. [Figure 56C] The graph shows the fold change from vehicle in the amount of TNF in the culture supernatant 24 hours after stimulation in human PBMC cultures from five donors cultured in the presence of GPR174 inhibitory compound 10 (1 μM); A2aR inhibitory compound (ZM, 0.2 μM); A2bR inhibitory compound MRS (2 μM); or a combination of compound 10 (1 μM) + 0.2 μM ZM + 2 μM MRS; or vehicle control (DMSO), as described in Example 22. [Figure 57] As described in Example 23, the graph shows the fold reduction of AREG+ cells in the presence of vehicle; GPR174 inhibitory compound 10 (1 μM); ZM-241385 (0.2 μM); or a combination of compound 10 (1 μM) and ZM (0.2 μM). [Figure 58] FIG. 1 is a schematic illustrating how both GPR174 and adenosine receptors respond to products of cellular stress and cell death, as described in Example 23. [Figure 59] Figure 59A graphically depicts the growth of CT26 tumors in individual wild-type (WT) mice inoculated with CT26.cl25 mouse colon carcinoma cells on day 0 and treated with anti-GITR antibodies on days 7 and 9, as described in Example 24. Figure 59B graphically depicts the growth of CT26 tumors in individual GPR174-KO mice inoculated with CT26.cl25 mouse colon carcinoma cells on day 0 and treated with anti-GITR antibodies on days 7 and 9, as described in Example 24. [Figure 60]10A-C graphically show the percent survival of tumor-bearing WT and GPR174-KO mice inoculated with CT26.cl25 mouse colon carcinoma cells on day 0 and treated with anti-GITR antibody on days 7 and 9, as described in Example 24, demonstrating that GPR174-KO mice had a higher percent survival (i.e., were euthanized significantly later) than WT mice (p=0.03, log-rank test). [Figure 61] Figure 61A graphically depicts the growth of CT26 tumors in individual WT mice inoculated with CT26.cl25 mouse colon carcinoma cells on day 0 and treated with anti-GITR antibodies on days 7, 9, and 14, as described in Example 24. Figure 61B graphically depicts the growth of tumors in individual GPR174-KO mice inoculated with CT26.cl25 mouse colon carcinoma cells on day 0 and treated with anti-GITR antibodies on days 7, 9, and 14, as described in Example 24. [Figure 62] Graph showing percent survival of tumor-bearing WT and GPR174-KO mice inoculated with CT26.cl25 mouse colon carcinoma cells on day 0 and treated with anti-GITR antibody on days 7, 9, and 14, as described in Example 24, demonstrating that GPR174-KO mice had a higher percent survival (i.e., were euthanized significantly later) than WT mice. [Figure 63]Figure 63A graphically depicts the growth of B16F10-Kb melanoma tumors in individual WT mice inoculated with B16F10-Kb cells on day 0 and treated with anti-GITR antibody on days 4 and 14, as described in Example 25. Figure 63B graphically depicts the growth of B16F10-Kb melanoma tumors in individual GPR174-KO mice inoculated with B16F10-Kb cells on day 0 and treated with anti-GITR antibody on days 4 and 14, as described in Example 25. Figure 63C graphically depicts the mean tumor volume of B16F10-Kb melanoma tumors in WT and GPR174-KO mice inoculated with B16F10-Kb cells on day 0 and treated with anti-GITR antibody on days 4 and 14, as described in Example 25, demonstrating that the mean tumor size was significantly smaller in GPR174KO mice on day 14 ("*" indicates p=0.01) and day 16 ("***" indicates p=0.00005). [Figure 64] 10A-10C are graphs showing percent survival of B16F10-Kb melanoma tumor-bearing WT and GPR174-KO mice inoculated with B16F10-Bb cells on day 0 and treated with anti-GITR antibody on days 4 and 14, as described in Example 25, demonstrating that GPR174-KO mice had a higher percent survival (i.e., were euthanized significantly later) than WT mice (p=0.006, log-rank test). [Figure 65] 1 graphically depicts IL-2 concentrations in cultures of stimulated human T cells in the presence or absence of PS liposomes (PSL) or a GPR174 inhibitor (compound 10, 3 μM), as described in Example 26. [Figure 66]Figure 66A graphically depicts the concentration of IL-2 in cultures of stimulated human T cells in the presence or absence of PS liposomes, tumor exosomes, or a GPR174 inhibitory compound (compound 10, 0.5 μM), as described in Example 26. Figure 66B graphically depicts the concentration of IFN-γ in cultures of stimulated human T cells in the presence or absence of PS liposomes, tumor exosomes, or a GPR174 inhibitory compound (compound 10, 0.5 μM), as described in Example 26. Figure 66C graphically depicts the concentration of TNF in cultures of stimulated human T cells in the presence or absence of PS liposomes, tumor exosomes, or a GPR174 inhibitory compound (compound 10, 0.5 μM), as described in Example 26. [Figure 67] 10A-10C are graphs illustrating IL-2 levels (fold change from vehicle) in cultures of stimulated WT or GPR174-KO mouse T cells in the presence or absence of PS liposomes (PSL) or a GPR174 inhibitor (compound 10, 1 μM), as described in Example 26. [Figure 68] Figure 68A graphically depicts the effects of A2a / A2b adenosine receptor antagonist ZM-241385 (0.1 μM), GPR174 inhibitory compound 10 (1 μM), or both compounds (ZM-241385 + compound 10) on IL-2 levels in the supernatants of cultured splenocytes isolated from n=3 WT mice after 2 days of stimulation with anti-CD3 and anti-CD28 antibodies, as described in Example 27. Figure 68B graphically depicts the effects of A2a / A2b adenosine receptor antagonist ZM-241385 (0.1 μM), GPR174 inhibitory compound 10 (1 μM), or both compounds (ZM-241385 + compound 10) on IL-2 levels in the supernatants of cultured splenocytes isolated from n=3 GPR174-KO mice after 2 days of stimulation with anti-CD3 and anti-CD28 antibodies, as described in Example 27. [Figure 69]10A-10C graphically show the effects of A2a / A2b adenosine receptor antagonist ZM-241385 (0.2 μM), GPR174 inhibitory compound 10 (0.3 μM) or GPR174 inhibitory compound 6 (0.3 μM), or ZM-241385 plus compound 10 or ZM-241385 plus compound 6 on IL-2 levels in the supernatant of cultured splenocytes isolated from three WT mice after two days of stimulation with anti-CD3 and anti-CD28 antibodies in the presence of the adenosine receptor agonist NECA (0.1 μM), as described in Example 27. [Figure 70A] FIG. 11 graphically shows the effects of the A2a / A2b adenosine receptor antagonist ZM-241385 (0.2 μM), the GPR174 inhibitory compound 10 (0.3 μM) or compound 6 (0.3 μM), or ZM-241385 alone or in combination with any of the GPR174 inhibitory compounds, on IL-2 levels in the supernatants of cultured splenocytes isolated from three WT mice after two days of stimulation with anti-CD3 and anti-CD28 antibodies in the presence of the adenosine receptor agonist NECA (0.1 μM), as described in Example 27. [Figure 70B] FIG. 11 graphically shows the effects of the A2a / A2b adenosine receptor antagonist ZM-241385 (0.2 μM), the GPR174 inhibitory compound 10 (0.3 μM) or compound 6 (0.3 μM), or ZM-241385 alone or in combination with any GPR174 inhibitory compound, on IL-2 levels in the supernatant of cultured splenocytes isolated from three GPR174-KO mice after two days of stimulation with anti-CD3 and anti-CD28 antibodies in the presence of the adenosine receptor agonist NECA (0.1 μM), as described in Example 27. [Figure 71] FIG. 11 is a graph showing the levels of IL-2 in the supernatants of human PBMCs stimulated with anti-CD3 and anti-CD28 in the presence or absence of GPR174 inhibitory compound 10 (1 μM); A2aR / A2bR inhibitory compound ZM-241385 (0.1 μM) or CD73 inhibitory compound APCP (10 μM), collected 24 hours after stimulation, as described in Example 28. [Figure 72A]10A-B graphically show the concentration of IL-2 in purified human T cells stimulated with CD3 / CD28 activators in the presence of PS liposomes (PS, 1 μM), the adenosine receptor agonist NECA (0.1 μM), or a combination of PS (1 μM) plus NECA (0.1 μM) compared to medium alone, as described in Example 29. [Figure 72B] A graph shows the concentration of IFN-γ (FIG. 72B) in purified human T cells stimulated with CD3 / CD28 activators in the presence of PS liposomes (PS, 1 μM), the adenosine receptor agonist NECA (0.1 μM), or a combination of PS (1 μM) plus NECA (0.1 μM) compared to medium alone, as described in Example 29. [Figure 72C] A graph shows the concentration of IFN-γ (Figure 72C) in purified human T cells stimulated with CD3 / C28 activators in the presence of PS liposomes (PS, 1 μM), the adenosine receptor agonist NECA (0.05 μM), or a combination of PS (1 μM) + NECA (0.05 μM) compared to medium alone, as described in Example 29. DETAILED DESCRIPTION OF THE INVENTION

[0317] Detailed Description As described in detail below, the present inventors have identified compounds that functionally interact with GPR174 and can inhibit one or more GPR174-mediated signaling pathways. The present inventors further characterized the signaling pathways activated by GPR174 and revealed the signaling profile for this receptor, including the Gs signaling pathway. The present inventors further demonstrated that combining GPR174 inhibition with adenosine 2a receptor (A2aR) and / or adenosine 2b receptor (A2bR) inhibition, or combining a GPR174 inhibitor with an inhibitor of ATP hydrolysis to adenosine by CD38, CD39, and CD73 (such as a CD73 inhibitor and / or a CD38 inhibitor and / or a CD39 inhibitor), and / or a Treg attenuator, resulted in synergistic induction of IFN-γ, IL-2, TNF, and GM-CSF production in human PBMCs. The present inventors further discovered that phosphatidylserine (PS) is an agonist of GPR174-mediated Gs signaling, and demonstrated that GPR174-mediated PS signaling is inhibited by multiple GPR174-inhibitory small molecule compounds with diverse chemical structures. As further demonstrated herein, GPR174 deficiency enhances antitumor immune responses in mice.

[0318] Based on these findings, the present disclosure relates to in vivo and in vitro methods of inhibiting the GPR174-mediated signaling pathway, alone or in combination with inhibition of ATP-adenosine A2aR and / or A2bR-mediated signaling (such as an A2aR antagonist and / or an A2bR antagonist), or in combination with inhibitors of GPR174-mediated signaling and inhibitors of ATP hydrolysis to adenosine by CD38, CD39, and CD73 (such as a CD73 inhibitor or a CD38 inhibitor or a CD39 inhibitor), and / or Treg attenuating agents, thereby stimulating an immune response in a mammalian subject, particularly a subject suffering from a condition such as cancer as described herein.

[0319] definition The term "G protein-coupled receptor" or "GPCR" or "GPR" refers to a transmembrane receptor that can transmit signals from the outside of a cell to the inside of a cell via the G protein pathway and / or the arrestin pathway. Hundreds of such receptors are known in the art; see, for example, Fredriksson et al., Mol. Pharmacol. 63:1256-1272, 2003, and Vassilatis, DK, Proc Natl Acad Sci USA 100:4903-4908 (2003), each of which is incorporated herein by reference. These references characterize human and mouse GPCRs based on sequence homology and function. Human GPCRs can be classified into five classes: secretin, rhodopsin, glutamate, frizzled / Tas2, and adhesion. Alternatively, receptors can be classified by their ligands, such as peptide hormones or small molecules (e.g., biogenic amines). Other classification schemes include the A-F classification, where class A represents receptors related to rhodopsin and adrenergic receptors, class B represents receptors related to calcitonin and parathyroid hormone receptors, class C represents receptors related to metabotropic receptors, and classes D-F represent receptors found in fungi and archaea.

[0320] The terms "G protein-coupled receptor 174," "GPR174," "FKSG79," or "GPCR17" refer to any naturally occurring form of the GPR174 protein, such as SEQ ID NO:1 as shown in FIG. 1, or a naturally occurring variant thereof, such as a variant having at least 90% identity (such as at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to SEQ ID NO:1. Preferred forms of GPR174 have the ability to signal through at least one G protein-coupled receptor pathway, such as Gs.

[0321] The term "G protein" refers to a heterotrimeric protein complex that transmits signals from activated GPCRs to intracellular effector molecules, such as enzymes and ion channels. G proteins are composed of Gα, Gβ, and Gγ subunits. The Gα subunit family includes Gq, Gi, Gs, and Gα12 / 13. G protein signaling pathways are named after the activated Gα subunits: Gαs, Gαi, Gαq, and Gα12 / 13. Heterotrimeric G proteins bind to activated GPCR proteins, i.e., GPCR proteins bound to a ligand or surrogate ligand. Upon binding to the GPCR protein, the Gα subunit exchanges its bound guanosine diphosphate (GDP) for guanosine-5'-triphosphate (GTP) and dissociates from the Gβ and Gγ subunits that typically associate in the heterodimeric complex. Upon dissociation, both the Gα-GTP-bound protein and the Gβγ complex can activate signaling pathways. The Gq family includes Gαq, Gα11, Gα14, and Gα15 / 16. The Gi family includes Gαi1-3, Gαo, Gαt, Gαgust, and Gαz. The Gs family includes Gαs and Gαolf. The G12 / 13 family includes Gα12 and Gα13.

[0322] The term "signal transduction pathway" refers to an intracellular response to a signal. In the present disclosure, a G protein signal transduction pathway refers to an intracellular response that occurs after a ligand, a surrogate ligand, or any other functional activator or inactivator binds to a G protein-coupled receptor, such as GPR174, activating a G protein-coupled pathway. G protein signal transduction pathways include the Gq, Gi, Gs, G12 / 13, and βγ pathways. An arrestin signal transduction pathway refers to an intracellular response that occurs after a ligand, a surrogate ligand, or any other functional activator or inactivator binds to a G protein-coupled receptor, such as GPR174, and activates an arrestin-mediated signal transduction pathway, such as the arrestin 1, arrestin 2, arrestin 3, and arrestin 4 signal transduction pathways.

[0323] The term "Gq signaling pathway" or "Gαq signaling pathway" refers to an intracellular signaling pathway activated by activated Gqα proteins. Gαq proteins are activated by exchanging GDP for GTP after binding to activated GPCR proteins. Activation of the Gq signaling pathway typically activates phospholipase Cβ isoform (PLCβ), which hydrolyzes phosphatidylinositol to produce diacylglycerol (DAG) and inositol 1,4,5-triphosphate (IP3). IP3 is soluble and diffuses through the cytoplasm, interacting with IP3 receptors on the endoplasmic reticulum, triggering calcium release and increasing intracellular calcium levels. Due to its hydrophobicity, DAG remains tethered to the inner leaflet of the plasma membrane, where it recruits protein kinase C (PKC), which is activated in conjunction with bound calcium ions. PKC then phosphorylates other proteins to regulate their function. Downstream effects, such as the expression of reporter genes regulated by appropriate transcription factors, can also be detected.

[0324] "Gi signaling pathway" or "Gαi signaling pathway" refers to the intracellular signaling pathway activated by activated Giα proteins. Gαi proteins are activated after binding to activated GPCR proteins. Gαi1-3, Gαo, and Gαz inhibit the activity of adenylyl cyclase (AC). The sensory Gi protein Gαt activates cyclic guanosine monophosphate (cGMP)-dependent phosphodiesterase, causing a decrease in intracellular cGMP, and Gαgust activates phospholipase C (PLC). All Gs except Gαz i Family members are sensitive to inhibition by pertussis toxin through adenosine diphosphate (ADP) ribosylation of their α subunits (Siehler, Biotechnology 3:471-83, 2008).

[0325] "Gs signaling pathway" or "Gαs signaling pathway" refers to an intracellular signaling pathway activated by activated Gsα protein. Gas proteins are activated after binding to activated GPCR proteins. The Gs family of G proteins (Gαs and Gαolf) activate ACs, which generate the second messenger 3',5'-cyclic adenosine monophosphate (cAMP).

[0326] The "G12 / 13 signaling pathway" or "Gα12 / 13 signaling pathway" refers to an intracellular signaling pathway activated by activated G12 / 13α proteins. Gα12 / 13 proteins are activated after binding to GPCR proteins. The G12 / 13 family of G proteins (Gα12 and Gα13) activates Rho guanine exchange factor (Rho-GEF) proteins. Rho-GEF proteins catalyze the exchange of GDP for GTP to activate RhoA. RhoA then activates Rho kinase, which further leads to the activation of activating transcription factor 2 (ATF2), resulting in a cellular response (Liu et al., Methods Mol. Biol. 237:145-9, 2004).

[0327] The "βγ signaling pathway" refers to an intracellular pathway activated by a free (i.e., uncoupled) βγ complex. The βγ complex can, for example, activate phospholipase A2, and when coupled to a muscarinic acetylcholine receptor, induce the opening of G protein-coupled inwardly rectifying potassium channels (GIRKs), activate L-type calcium channels, and activate PLC, thereby initiating the phospholipase C pathway.

[0328] "Arrestin signaling pathway" refers to an intracellular signaling pathway activated by arrestin 1, arrestin 2, arrestin 3, or arrestin 4. Arrestins are involved in signaling through ERK, JNK, p38, Akt, PI3 kinase, and RhoA (DeWire et al., Annu. Rev. Physiol. 69:483-510, 2007).

[0329] The term "contacting" is used interchangeably herein with combined with, added to, mixed with, introduced into, passed through, incubated with, flowed over, etc. For clarity, the phrase "contacting a cell" includes introducing a compound into a mammal (e.g., orally, into plasma, or intramuscularly) such that the compound contacts the cells of the mammal in vivo.

[0330] The term "GPR174-mediated signaling pathway" means a signaling pathway whose activity is regulated at least in part through GPR174.

[0331] As used herein, the term "modulating at least one GPR174-mediated signal transduction pathway" refers to modulating (activating or inhibiting) at least the Gs signal transduction pathway that is functional in cells that express GPR174 by contacting the cells with a compound that functionally interacts with GPR174.

[0332] A "modulator" is a compound that functionally interacts with a GPCR and affects GPCR-mediated signaling activity, either alone or by altering the ability of another compound to affect GPCR-mediated signaling activity. Modulators include activators and inhibitors of the GPCR-mediated signaling pathway.

[0333] An "activator" is a compound that increases GPCR-mediated signal transduction in a signal transduction pathway. An activator can directly activate the receptor (e.g., an agonist) or increase the activation of the receptor by another compound (e.g., a positive allosteric modulator). An activator can bind to the receptor and increase, increase, open, activate, promote, enhance activation, sensitize, stimulate, induce receptor conformation to an active state, or upregulate receptor-protein activity. Activators include agonists, partial agonists, and positive allosteric modulators.

[0334] An "agonist," which may be a full agonist or a partial agonist, binds to a GPCR, activates the receptor, and typically initiates a response via activation of a G protein. An agonist increases receptor activity compared to normal baseline levels. A compound is considered a receptor agonist if it increases GPCR-mediated signaling pathway activity by at least 20% above an appropriate control (e.g., background or basal activity) at concentrations up to 50 μM. A full agonist is always an activator. A partial agonist has lower intrinsic activity than a full agonist, but increases receptor activity compared to basal activity. A partial agonist may act as a functional inhibitor in the presence of a full agonist.

[0335] A "positive allosteric modulator" or "PAM" is a compound that binds to a GPCR at a site distinct from the ligand binding site and increases GPCR-mediated signaling activity in response to an agonist.

[0336] An "inhibitor" is a compound that reduces GPCR-mediated signal transduction in a signal transduction pathway. An inhibitor is a compound that functionally interacts with GPCR and partially or completely blocks, reduces, suppresses, delays activation, inactivates, antagonizes, desensitizes, induces the receptor's conformation to an inactive conformation, blocks the ability of another compound (e.g., an endogenous agonist ligand) to interact with the receptor, or otherwise downregulates the activity of the receptor. An inhibitor can reduce the basal activity of the receptor (e.g., an inverse agonist) or can block or reduce the activity of another compound (e.g., a partial agonist or antagonist). Inhibitors include antagonists, inverse agonists, partial agonists, partial inverse agonists, and negative allosteric modulators. Inhibitors do not include compounds that act only by reducing the expression of receptor nucleic acid or protein.

[0337] An "antagonist" is an inhibitor that typically binds to GPCR at the same site as a ligand or agonist. By itself, an antagonist does not activate or inhibit receptor-mediated signaling activity, and does not change receptor activity from basal levels. However, an antagonist can inhibit or block receptor activation in the presence of an agonist or ligand, or inhibit or block receptor inhibition in the presence of an inverse agonist. A compound is considered to be a receptor antagonist or allosteric modulator if it has activity in a CRA assay at concentrations up to 50 μM, and increases GPCR receptor-mediated signaling pathway activity by less than 20% over an appropriate control (e.g., background or basal), or decreases GPCR-mediated signaling pathway activity by less than 10% compared to an appropriate control (e.g., background or basal). Furthermore, a compound that inhibits the effect of an agonist or inverse agonist can be considered an antagonist.

[0338] An "inverse agonist" can be a full inverse agonist or a partial inverse agonist, and is an inhibitor that binds to a receptor and reduces the basal signal transduction of a GPCR-mediated signal transduction pathway. A compound is considered to be an inverse agonist if it reduces the basal signal transduction of a GPCR-mediated signal transduction pathway by at least 10% at a concentration of up to 50 μM. A partial inverse agonist has a lower intrinsic inhibitory activity than a full inverse agonist.

[0339] A "negative allosteric modulator" is a compound that binds to a receptor at a site distinct from the ligand binding site and decreases GPCR-mediated signaling activity in response to an agonist.

[0340] A "partial agonist" can be a functional activator or a functional inhibitor depending on the presence or absence of a full agonist in a given biological environment. A partial agonist binds to a receptor and increases receptor activity compared to the basal level of activity, but has only partial effectiveness compared to a full agonist. For example, in the presence of a full agonist, a partial agonist can reduce receptor activation, thereby acting as a functional inhibitor. In the absence of a full agonist, a partial agonist can increase receptor activation, thereby acting as a functional activator.

[0341] A "partial inverse agonist" can be a functional activator or a functional inhibitor depending on the presence or absence of a full inverse agonist in a given biological environment. A partial inverse agonist binds to a receptor and reduces receptor activity compared to the basal level of activity, but has only partial effectiveness compared to a full inverse agonist. For example, in the presence of a full inverse agonist, a partial inverse agonist can increase receptor activation, thereby acting as a functional activator. In the absence of a full inverse agonist, a partial inverse agonist can decrease receptor activation, thereby acting as a functional inhibitor.

[0342] As used herein, the term "basal level of activity" or "basal signaling activity" or "baseline activity" refers to the level of GPCR-mediated signaling activity in the absence of a modulator compound. In one embodiment, the basal level of GPR174-mediated signaling pathway activity is established with reference to a particular cell line, which is known to have functional or defective GPR174 activity. In another embodiment, the basal level of GPR174-mediated signaling pathway activity is established with reference to cells that overexpress GPR174 (e.g., as described in Examples 2, 3, and 4 herein). In some embodiments, the basal level of GPR174-mediated signaling pathway activity is established with reference to cells known to interact with or contain an endogenous GPR174 ligand, or with reference to cells known not to contain an endogenous GPR174 ligand, assuming that an endogenous GPR174 ligand is later identified.

[0343] A "ligand" is a compound that binds to a receptor and modulates the activity of the receptor.

[0344] An "endogenous ligand" is an endogenous entity (eg, a molecule, peptide, or ion) that modulates a receptor in the cell, tissue, or organism of origin.

[0345] A "surrogate ligand" is a non-endogenous molecule that binds to a receptor and modulates its activity. A surrogate ligand may be naturally occurring or synthetic. A surrogate ligand can activate or inhibit the activity of a receptor. In one aspect, a surrogate ligand is a small molecule. In a further aspect, a surrogate ligand is a small organic molecule.

[0346] A "biased ligand" is a compound that binds to a GPCR and selectively modulates (more than two-fold) the activity of one GPCR-mediated signaling pathway (e.g., the Gq, Gi, Gs, G12 / 13, βγ, or arrestin signaling pathway) relative to other GPCR-mediated signaling pathways.

[0347] As used herein, the term "apparent binding affinity" refers to the observed change in the binding affinity of a compound to GPR174 in the presence of an additional compound that binds to GPR174, compared to the binding affinity observed for the compound to GPR174 in the absence of the additional compound. In some embodiments, the compound and the additional compound compete for binding at the same site on GPR174. While the binding affinity of each compound to GPR174 does not change, determining the binding affinity of one compound to GPR174 in the presence of an additional compound that binds to GPR174 may result in a decrease in the measured binding affinity of the compound to GPR174. In some embodiments, the two compounds do not bind to the same site, and allosteric binding of one compound to GPR174 may result in a decrease or increase in the binding affinity of the other compound to GPR174. This may be due to a conformational change in GPR174 caused by the binding of one of the compounds to GPR174, which affects the binding site of the other compound.

[0348] A compound is "specific for" GPR174 if it can modulate (e.g., inhibit) GPR174 activity but does not have similar activity at other proteins, such as other G protein-coupled receptors, for example, compared to a reference panel of proteins (e.g., including GPCRs other than GPR174, and optionally including ion channels and / or other types of transporters). In one example, the reference panel comprises the following receptors: muscarinic M1, CCRL2, CMKOR1, GPR3, GPR4, GPR12, GPR17, GPR18, GPR19, GPR20, GPR21, GPR22, GPR25, GPR26, GPR27, GPR31, GPR32, GPR34, GPR37, GPR37L1, GPR39, GPR43, GPR45, GPR48, GPR50, GPR52, GPR61, GPR62, GPR63, GPR65, GPR68, GPR78, GPR80, GPR 83, GPR85, GPR87, GPR88, GPR101, GPR132, GPR135, GPR139, GPR141, GPR146, GPR148, GPR149, GPR150, GPR151, GPR152, GPR153, GPR160, GPR161, GPR162, GPR173, GPR182, GPR183, LGR5, LGR6, MAS1, MRGD, MRGE, MRGF, MRGI4, OPN3, OPN4, OPN5, P2Y8, P2Y10, TAAR6, and TAAR8.

[0349] In another example, the reference panel includes the following receptors: human adenosine A1; human adenosine A2A; rat adrenergic alpha 1A; rat adrenergic alpha 1B; human adrenergic alpha 2A; human adrenergic beta 1; human adrenergic beta 2; rat calcium channel L-type dihydropyridine; human dopamine D1; human dopamine D2; and human G protein-coupled receptor, GPR103. In another example, the reference panel includes the following receptors: rat GABA A; rat glutamate NMDA; human histamine H1; human muscarinic M2; human nicotinic acetylcholine alpha 1; human opiate μ (OP3, MOP); human potassium channel [K ATP]; human potassium channel hERG; human sigma σ1; rat sigma σ2; rat sodium channel, site 2; and human transporter, norepinephrine.

[0350] In certain embodiments, compounds specific for GPR174 have 2-fold, 5-fold, 10-fold, 25-fold, 50-fold, 100-fold, 500-fold, 1000-fold, 5000-fold, or 10,000-fold greater activity in inhibiting GPR174 activity compared to other G protein-coupled receptors, e.g., other GPCRs in one or more of the above reference panels.

[0351] In certain embodiments, compounds specific for GPR174 have 2-fold, 5-fold, 10-fold, 25-fold, 50-fold, 100-fold, 500-fold, 1000-fold, 5000-fold, or 10,000-fold greater activity in Gs signaling assays performed in cells expressing GPR174 compared to Gs signaling activity in cells expressing other GPCRs, such as other GPCRs in each of the above reference panels.

[0352] "Agency-approved compound" refers to a compound approved for clinical use for human or veterinary purposes by the U.S. Food and Drug Administration (FDA) or a similar government regulatory agency (e.g., the European Medicines Agency (EMA), Health Canada, or the Ministry of Health, Labor and Welfare of Japan) before November 4, 2016.

[0353] A compound that is "not approved for use in a disease associated with GPR174" refers to a compound that has not been approved for therapeutic use by the FDA or a similar government regulatory agency (e.g., EMA, Health Canada, and the Ministry of Health, Labor and Welfare of Japan) before November 4, 2016, for an indication associated with GPR174 activity (e.g., any of those described herein, e.g., cancer or a nervous system disease or disorder).

[0354] A compound "not approved as targeting GPR174" means a compound that has not been shown to target GPR174 in pharmacology data submitted to the U.S. FDA or similar government regulatory agencies (e.g., EMA, Health Canada, and the Japanese Ministry of Health, Labor and Welfare) prior to November 4, 2016. Indications for such targeting are often found on the labeling of approved drugs.

[0355] As used herein, the term "compound" or grammatical equivalents refers to natural or synthetic molecules, such as proteins; antibodies, oligopeptides (e.g., about 5 to about 25 amino acids in length, e.g., about 10 to 20 or 12 to 18 amino acids in length, e.g., 12, 15, or 18 amino acids in length); nucleotides that inhibit GPR174 expression (e.g., inhibitory RNA), small chemical compounds, such as small organic, organometallic, or inorganic molecules; polysaccharides; oligonucleotides; lipids; and fatty acids. Compounds can be included in libraries of compounds, such as combinatorial, synthetic, natural, heterocyclic, drug-like, lead-like, organic, inorganic, non-randomized, or randomized libraries, providing a sufficient range of diversity, or can be focused or targeted collections of the above compounds. Compounds are optionally linked to fusion partners, such as targeting compounds, rescue compounds, dimerization compounds, stabilizing compounds, addressable compounds, and other functional moieties. Traditionally, new chemical entities with useful properties are generated by identifying a compound (called a "lead compound") that has some desirable property or activity, e.g., inhibitory activity, generating variants of the lead compound, and evaluating the properties and activities of those variant compounds. Often, high-throughput screening ("HTS") methods are used for such analysis.

[0356] The terms "small molecule," "small organic molecule," and "small inorganic molecule" refer to molecules (whether organic, organometallic, or inorganic), organic molecules, and inorganic molecules, respectively, that are either natural or synthetic and have a molecular weight greater than about 50 Da and less than about 2500 Da. Small organic (for example) molecules can be less than about 2000 Da, between about 100 Da and about 1000 Da, or between about 100 and about 600 Da, or between about 200 and 500 Da.

[0357] The term "a compound pre-determined to functionally interact with GPR174" refers to a compound that has been determined to modulate a GPR174-mediated signal transduction pathway by functionally interacting with GPR174 prior to contacting a cell expressing GPR174 with the compound or administering the compound to a subject. This pre-determination, in either case, is performed prior to contacting a cell expressing GPR174 with the compound or administering the compound to a subject. (i) directly obtaining the knowledge that a compound functionally interacts with GPR174 by physically demonstrating that the compound functionally interacts with GPR174 by performing an assay that measures functional interaction with GPR174 or by carrying out an assay that measures GPR174-mediated signaling activity; or (ii) Indirectly gaining knowledge that a compound functionally interacts with GPR174 by receiving or obtaining from another person or source knowledge or information regarding results from another person that physically demonstrate such interaction using an assay that measures the functional interaction of a compound with GPR174 or an assay that measures GPR174-mediated signaling activity (e.g., obtained, for example, from a publication in a scientific journal or in a published patent application or by reading prescribing information, or obtained through private channels (e.g., personal conversation)) that physically demonstrate such interaction using an assay that measures the functional interaction of a compound with GPR174 or an assay that measures GPR174-mediated signaling activity). In one embodiment, a compound is pre-determined to functionally interact with human GPR174 by directly or indirectly finding that the compound functionally interacts with human GPR174, as set forth as SEQ ID NO:1, or with a naturally occurring variant of GPR174 having at least 95% identity to SEQ ID NO:1.

[0358] A "therapeutically effective amount" means the amount for which it is administered that produces the desired effect, e.g., amelioration or delay of at least one symptom associated with the disease or condition being treated. The exact dose will depend on the purpose of the treatment and can be ascertained by one skilled in the art using known techniques (see, e.g., Lieberman, Pharmaceutical Dosage Forms (vols. 1-3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); and Pickar, Dosage Calculations (1999)).

[0359] "Substantially pure" or "isolated" refers to a compound (e.g., a polypeptide or conjugate) separated from other chemical components. Typically, a compound is substantially pure when it is at least 30% by weight free from other components. In certain embodiments, a preparation is at least 50%, 60%, 75%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% free by weight from other components. A purified polypeptide can be obtained, for example, by expression of a recombinant polynucleotide encoding such a polypeptide or by chemically synthesizing the polypeptide. Purity can be measured by any appropriate method, for example, column chromatography, polyacrylamide gel electrophoresis, or by HPLC analysis.

[0360] In the context of a naturally occurring compound, the term "isolated" refers to one that has been altered or removed from the natural state (eg, through human intervention).

[0361] The term "mammal" includes all mammals, including, but not limited to, humans, non-human primates, dogs, cats, horses, sheep, goats, cows, rabbits, pigs, and rodents.

[0362] As used herein, and as is well understood in the art, "treating" a disease, disorder, or condition, "treatment" of a disease, disorder, or condition (e.g., a condition described herein, such as an inflammatory condition), or "therapy" is an approach to obtain a beneficial or desired result, such as a clinical result, and can be carried out for prophylaxis or during the course of clinical pathology. Beneficial or desired results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, whether detectable or undetectable; reduction in the severity of the disease, disorder, or condition; reduced likelihood of developing the disease, disorder, or condition; stabilized (i.e., non-worsening) state of the disease, disorder, or condition; inhibiting the onset or spread of the disease, disorder, or condition; delaying or slowing the progression of the disease, disorder, or condition; improvement or palliation of the disease, disorder, or condition; and remission (whether partial or complete). To "ameliorate" a disease, disorder, or condition means to lessen the severity and / or undesirable clinical findings of the disease, disorder, or condition and / or slow or prolong the time course of progression compared to the severity or time course in the absence of treatment.

[0363] As used herein, the term "subject" or "patient" refers to any organism to which a compound or composition according to the present disclosure can be administered, for example, for experimental, diagnostic, prophylactic, and / or therapeutic purposes. A subject to be treated with a compound or composition described herein can be one who has been diagnosed by a physician as having a disease, disorder, or condition described herein, or one who is at risk for developing a disease, disorder, or condition described herein. Diagnosis can be performed by any technique or method known in the art. One skilled in the art will understand that a subject can be diagnosed with a disease, disorder, or condition using standard tests or examinations, or can be identified as a high-risk subject due to the presence of one or more risk factors without examination. Typical subjects include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and humans).

[0364] As used herein, the term "neoplasm" refers to any new and abnormal growth of cells, especially one in which the division and proliferation of cells is uncontrolled and progressive. Neoplasms can be non-malignant (i.e., benign) or malignant.

[0365] As used herein, the term "tumor" refers to neoplasms, including solid and liquid (i.e., blood) neoplasms, as well as benign and malignant neoplasms, including primary and / or metastatic neoplasms.

[0366] As used herein, the term "alkanoyl" refers to a group having the structure -C(O)-R, where R is alkyl. The alkanoyl may be unsubstituted or substituted as described for alkyl (e.g., optionally substituted alkanoyl). The suffix "oyl" may be used to define other groups having the structure -C(O)-R. For example, in an alkenoyl group, R is alkenyl; in an alknynoyl group, R is alkynyl; in a cycloalkanoyl group, R is cycloalkyl; in a cycloalkenoyl group, R is cycloalkenyl; and in a cycloalkynyl group, R is cycloalkynyl (all groups are as defined herein). Additionally, a group defined with the suffix "oyl" may be further used to define a group having the structure -O-C(O)-R' by adding the suffix "oxy," e.g., when R' is alkyl, the group is an "alkanoyloxy." For example, in an alkenoyloxy group, R' is alkenyl, in an alknynoyloxy group, R' is alkynyl, in a cycloalkanoyloxy group, R' is cycloalkanyl, in a cycloalkenoyloxy group, R' is cycloalkenyl, and in a cycloalkynoyloxy group, R' is cycloalkynyl (all groups are as defined herein). Each of these groups can be unsubstituted or substituted (e.g., optionally substituted) as described for each respective group.

[0367] The term "alkenyl" as used herein refers to a straight- or branched-chain monovalent substituent containing one or two carbon-carbon double bonds and, if unsubstituted, containing only C and H. Unless otherwise specified, an alkenyl group may contain 2, 3, 4, 5, or 6 carbon atoms, excluding the carbon atoms of any substituents, if any. Non-limiting examples of alkenyl groups include ethenyl, prop-1-enyl, prop-2-enyl, 1-methylethenyl, but-1-enyl, but-2-enyl, but-3-enyl, 1-methylprop-1-enyl, 2-methylprop-1-enyl, and 1-methylprop-2-enyl. Alkenyl can be unsubstituted or substituted (e.g., optionally substituted alkenyl) as described for alkyl.

[0368] The term "alkenylene," as used herein, refers to a straight- or branched-chain divalent substituent containing one or two carbon-carbon double bonds and, if unsubstituted, containing only C and H. Unless otherwise specified, an alkenylene group may contain 2, 3, 4, 5, or 6 carbon atoms, excluding carbon atoms of any substituents, if any. Non-limiting examples of alkenylene groups include ethene-1,1-diyl; ethene-1,2-diyl; prop-1-ene-1,1-diyl, prop-2-ene-1,1-diyl; prop-1-ene-1,2-diyl, prop-1-ene-1,3-diyl; prop-2-ene-1,1-diyl; prop-2-ene-1,2-diyl; but-1-ene-1,1-diyl; but-1-ene-1,2-diyl; but-1-ene-1,3-diyl; but-1-ene-1,4-diyl; but-2-ene-1,1-diyl; but-2-ene-1,2-diyl; but-2-ene-1,3-diyl; but-2-ene-1,4-diyl; but- buta-1,3-diene-1,4-diyl; buta-1,3-diene-1,1-diyl; buta-1,3-diene-1,2-diyl; buta-1,3-diene-1,2-diyl; buta-1,3-diene-1,4-diyl; buta-1,3-diene-1,1-diyl; buta-1,3-diene-1,2-diyl; buta-1,3-diene-1,3-diyl; buta-1,3-diene-1,4-diyl; buta-1,3-diene-2,3-diyl; buta-2,3-diene-1,1-diyl; and buta-2,3-diene-1,2-diyl. The alkenylene can be unsubstituted or substituted as described for alkylene (eg, optionally substituted alkenylene).

[0369] The term "alkoxy" refers to a chemical substituent of formula -OR, where R is an optionally substituted alkyl group (e.g., an optionally substituted C1-C6 alkyl group). Substituted alkoxy groups can have 1, 2, 3, 4, 5, or 6 substituents as defined herein. Similarly, the term "arylalkoxy" refers to a chemical substituent of formula -OR, where R is an optionally substituted arylalkyl...

Claims

1. below: i. Structure: Compound 10 having the formula: ii. Structure: Compound 6 having the formula: iii. Structure: Compound 11 having the formula: iv. Structure: Compound 20 having the formula A pharmaceutical composition for treating cancer in a patient comprising a therapeutically effective amount of a GPR174 inhibitor selected from the group consisting of: i. ZM-241385; ii. SCH-58261; iii. PBF-509; iv. MRS-1754; and v. Anti-GITR antibody and administered in combination with at least one additional agent selected from the group consisting of: A pharmaceutical composition, wherein the pharmaceutical composition and the at least one additional agent are administered simultaneously or sequentially in any order, provided that the effect of the first administered inhibitor or agent is still present at the time of the second administered inhibitor or agent.

2. The pharmaceutical composition of claim 1, wherein the GPR174 inhibitor inhibits PS-dependent or LysoPS-dependent activation of GPR174 signaling in cells expressing GPR174 by at least 25%.

3. The pharmaceutical composition of claim 1, wherein the GPR174 inhibitor stimulates an immune response in a patient by inhibiting GPR174G-α-s signaling.

4. The pharmaceutical composition of claim 3, wherein the stimulated immune response comprises an NK cell-mediated immune response or an anti-tumor cytotoxic T cell-mediated immune response.

5. Human peripheral blood mononuclear cells (PBMCs) were cultured as follows: i. Structure: Compound 10 having the formula: ii. Structure: Compound 6 having the formula: iii. Structure: Compound 11 having the formula: iv. Structure: Compound 20 having the formula and at least one of an adenosine A2A (A2A) receptor antagonist, an adenosine A2B (A2B) receptor antagonist, or a combination thereof, wherein the inhibitor of GPR174 signaling is selected from the group consisting of: A2A receptor antagonists and A2B receptor antagonists i. ZM-241385; ii. SCH-58261; iii. PBF-509; and iv. MRS-1754 The method of claim 1, wherein the

6. The method of claim 5, wherein the PBMCs comprise T cells or NK cells.

7. The method of claim 5 or 6, wherein the level of at least one of the Th1 cytokines IFN-γ, IL-2, TNF, or GM-CSF is increased by at least 20%.

8. A pharmaceutical composition for treating cancer comprising a combination of a GPR174 inhibitor and at least one of an adenosine A2A (A2A) receptor antagonist, an adenosine A2B (A2B) receptor antagonist, and optionally a pharma- ceutically acceptable excipient, The GPR174 inhibitor is: i. Structure: Compound 10 having the formula: ii. Structure: Compound 6 having the formula: iii. Structure: Compound 11 having the formula: iv. Structure: Compound 20 having the formula is selected from the group consisting of A2A receptor antagonists and A2B receptor antagonists i. ZM-241385; ii. SCH-58261; iii. PBF-509; and iv. MRS-1754 A pharmaceutical composition selected from the group consisting of:

9. 1. A pharmaceutical composition for enhancing an anti-tumor immune response in a subject currently undergoing treatment with at least one of an A2aR antagonist, an A2bR antagonist, and a Treg attenuating agent, comprising an effective amount of a GPR174 inhibitor, wherein the GPR174 inhibitor is: i. Structure: Compound 10 having the formula: ii. Structure: Compound 6 having the formula: iii. Structure: Compound 11 having the formula: iv. Structure: Compound 20 having the formula is selected from the group consisting of A2A receptor antagonists, A2B receptor antagonists, and Treg attenuators i. ZM-241385; ii. SCH-58261; iii. PBF-509; iv. MRS-1754; and v. Anti-GITR antibody A pharmaceutical composition selected from the group consisting of:

10. The pharmaceutical composition of claim 9, wherein the GPR174 inhibitor inhibits PS-dependent or LysoPS-dependent activation of GPR174 signaling in cells expressing GPR174 by at least 25%.

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