Compounds, compositions, and methods for the treatment of diseases

Abstract

To provide a compound for treating cancer in a subject, particularly liver cancer.SOLUTION: The present invention provides a compound of Formula (I), or a pharmaceutically acceptable salt or stereoisomer thereof (where Z represents S or O; B1 represents a pyrimidinyl nucleobase; B2 represents a purinyl nucleobase; X1 and X2 independently represent O or S; Y1 and Y2 independently represent O, S, or N; L1 and L2 independently represent the absence or C1-C6 alkyl or the like; R1 and R2 independently represent H, halo, -CN, C1-C20 alkyl or the like; and R3 and R4 independently represent H, C1-C20 alkyl or the like).SELECTED DRAWING: Figure 1A

Description

[Technical Field]

[0001] Related Applications This application is a continuation of U.S. Provisional Patent Application No. 62 / 359,039, filed July 6, 2016; U.S. Provisional Patent Application No. 62 / 363,118, filed July 15, 2016; U.S. Provisional Patent Application No. 62 / 403,530, filed October 3, 2016; U.S. Provisional Patent Application No. 62 / 411,424, filed October 21, 2016; U.S. Provisional Patent Application No. 62 / 411,424, filed January 9, 2017; This application claims the benefit of priority to U.S. Provisional Patent Application No. 62 / 444,141, filed February 23, 2017, U.S. Provisional Patent Application No. 62 / 462,679, filed March 13, 2017, and U.S. Provisional Patent Application No. 62 / 470,746, filed May 19, 2017, the contents of each of which are hereby incorporated by reference in their entirety.

[0002] The present disclosure relates to compounds and compositions that activate the innate immune defense system and induce expression of pattern recognition receptors in a host, and methods of use for the treatment of proliferative diseases (e.g., cancer). [Background technology]

[0003] A key feature of the innate immune system is the recognition and elimination of foreign substances. Identification of these pathogenic invaders occurs through host recognition of evolutionarily conserved microbial structures known as pathogen-associated molecular patterns (PAMPs) (Jensen, S. and Thomsen, ARJ Virol (2012) 86:2900-2910). These PAMPs can be widely shared by multiple microbial species and include a wide range of molecular structures, such as nucleic acids, lipopolysaccharides, and glycoproteins, that are critical for the survival and / or pathogenicity of these microbial species. Host recognition can occur through multiple pathways, such as the activation of pattern recognition receptors (PRRs), which ultimately leads to downstream signaling events and ultimately to an enhanced immune response.

[0004] To date, several PRRs that serve as sensors of pathogenic infection have been identified. For example, the retinoic acid-inducible gene I (RIG-I) protein is an RNA helicase that also functions as a sensor of microbial RNA. RIG-I is a key factor in host recognition of RNA viruses from various different virus families, including Flaviviridae (e.g., West Nile virus, Hepatitis C virus, Japanese encephalitis virus, and Dengue virus), Paramyxoviridae (e.g., Sendai virus, Newcastle disease virus, respiratory syncytial virus, and measles virus), Rhabdoviridae (e.g., rabies virus), Orthomyxoviridae (e.g., influenza A virus, influenza B virus), and Arenaviridae (e.g., Lassa virus), as well as a biomarker for predicting the prognosis of certain types of cancer, such as hepatocellular carcinoma (Hou, J. et al., Cancer Cell (2014) 25:49-63). Stimulator of interferon genes (STING) is a cytoplasmic adaptor protein that activates the TBK1-IRF3 signaling complex, resulting in the induction of type I interferons (IFN-β and IFN-α) and other immune pathway proteins. Other PRRs also play a role in sensing microbial-derived nucleic acids, including NOD2, LGP2, MDA5, and several Toll-like receptors (TLRs), which are expressed on the cell surface and in endosomal compartments.

[0005] Recent publications have highlighted the importance of RIG-I and STING as mediators of innate and adaptive immunity, and RIG-I and STING agonists have been recognized as immuno-oncology drugs in cancer therapy (Li, XY et al., Mol Cell Oncol (2014) 1:e968016; Woo, SR Trends in Immunol (2015) 36:250-256). In particular, RIG-I is involved in regulating fundamental cellular processes, such as hematopoietic proliferation and differentiation, maintenance of leukemic stemness, and hepatocellular carcinoma tumorigenesis, indicating that RIG-I plays an essential function as a tumor suppressor. Importantly, the cytosolic DNA-sensing STING pathway has been shown to play a key mechanistic role in innate immune sensing and drive type I IFN production in cancer and in the context of immuno-oncology applications, including therapeutic and diagnostic agents. [Prior art documents] [Non-patent literature]

[0006] [Non-Patent Document 1] Jensen, S. and Thomsen, ARJ Virol (2012) 86:2900-2910 [Non-patent document 2] Hou, J. et al, Cancer Cell (2014) 25:49-63 [Non-patent document 3] Li, XYet al, Mol Cell Oncol (2014)1:e968016 [Non-patent document 4] Woo,SRTrends in Immunol(2015)36:250-256 Summary of the Invention

[0007] Described herein are cyclic dinucleotide compounds, compositions comprising cyclic dinucleotide compounds, and related methods of use.

[0008] In one aspect, the present disclosure provides a compound of formula (I): [ka] (wherein Z is either S or O, and B 1 and B 2 each of X is independently a purinyl nucleobase or a pyrimidinyl nucleobase; 1 and X 2 each independently is O or S, and Y 1 and Y 2 each independently represents O, S, or NR 5 and L 1 and L 2 each independently is absent, C1-C6 alkyl, or C1-C6 heteroalkyl, where alkyl and heteroalkyl are each optionally selected from R 6 is substituted with R 1 and R 2 each independently represents hydrogen, halo, —CN, C1-C 20 alkyl (e.g., C1-C6 alkyl), or OR 7 and R 3 and R 4 each independently represents hydrogen, C1 to C 20 Alkyl (e.g., C1-C6 alkyl), C1-C 20 Heteroalkyl (e.g., C1-C6 heteroalkyl), OC(O)OC1-C 20 alkyl (e.g., C1-C6 alkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each optionally contain one or more R 8 is substituted with R 5 is hydrogen or C1-C 20 alkyl (e.g., C1-C6 alkyl), and R 6 Halo, -CN, C1~C 20 alkyl (e.g., C1-C6 alkyl), OR 7, oxo, cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl are each optionally joined by one or more R 9 It is substituted with R 7 is hydrogen, C1 to C 20 alkyl (e.g., C1-C6 alkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each optionally contain one or more R 9 and each R 8 are C1 to C independently 20 Alkyl (e.g., C1-C6 alkyl), C1-C 20 Heteroalkyl, C(O)-C1-C 20 Alkyl, OC(O)-C1-C 20 Alkyl (e.g., C1-C6 alkyl), C(O)O-C1-C 20 Alkyl (e.g., C1-C6 alkyl), OC(O)O-C1-C 20 Alkyl (e.g., C1-C6 alkyl), C(O)N(R 5 )-C1~C 20 Alkyl (e.g., C1-C6 alkyl), N(R 5 )C(O)-C1~C 20 Alkyl (e.g., C1-C6 alkyl), OC(O)N(R 5 )-C1~C 20 Alkyl (e.g., C1-C6 alkyl), O-aryl, O-heteroaryl, C(O)-aryl, C(O)-heteroaryl, OC(O)-aryl, C(O)O-aryl, OC(O)-heteroaryl, C(O)O-heteroaryl, C(O)O-aryl, C(O)O-heteroaryl, C(O)N(R 5 )-aryl, C(O)N(R 5 )-heteroaryl, N(R 5 )C(O)-aryl, N(R 5 )2C(O)-alkyl, or N(R 5 )C(O)-heteroaryl, S(O)N(R 5)-aryl, wherein alkyl, heteroalkyl, aryl, and heteroaryl are each optionally joined by one or more R 9 is substituted with R 9 are each independently C1 to C 20 Alkyl, O-C1-C 20 Alkyl, C1-C 20 heteroalkyl, halo, —CN, OH, oxo, aryl, heteroaryl, O-aryl, or O-heteroaryl), or a pharmaceutically acceptable salt or stereoisomer thereof.

[0009] In some embodiments, the compound has formula (Ia): [ka] (In the formula, B 1 and B 2 each of X is independently a purinyl nucleobase or a pyrimidinyl nucleobase; 1 and X 2 is independently O or S, and Y 1 and Y 2 each independently represents O, S, or NR 5 and L 1 and L 2 each is independently absent, C1-C6 alkyl, or C1-C6 heteroalkyl, where alkyl and heteroalkyl are each optionally selected from R 6 is substituted with R 1 and R 2 each independently represents hydrogen, halo, —CN, C1-C 20 alkyl (e.g., C1-C6 alkyl), or OR 7 and R 3 and R 4 each independently represents hydrogen, C1 to C 20 Alkyl (e.g., C1-C6 alkyl), C1-C 20heteroalkyl (e.g., C1-C6 heteroalkyl), cycloalkyl, heterocyclyl, aryl, wherein alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each optionally contain one or more R 8 is substituted with R 5 is hydrogen or C1-C 20 alkyl (e.g., C1-C6 alkyl), and R 6 Halo, -CN, C1~C 20 alkyl (e.g., C1-C6 alkyl), OR 7 , oxo, cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl are each optionally joined by one or more R 9 is substituted with R 7 is hydrogen, C1 to C 20 alkyl (e.g., C1-C6 alkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, where alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl are optionally joined by one or more R 9 is substituted with R 8 are each independently C1 to C 20 Alkyl (e.g., C1-C6 alkyl), C1-C 20 Heteroalkyl, C(O)-C1-C 20 Alkyl, OC(O)-C1-C 20 Alkyl, C(O)OC1~C 20 Alkyl, OC(O)OC1~C 20 Alkyl, C(O)N(R 5 )-C1~C 20 Alkyl, N(R 5 )C(O))-C1~C 20 Alkyl, OC(O)N(R 5 )-C1~C 20 Alkyl, O-aryl, O-heteroaryl, C(O)-aryl, C(O)-heteroaryl, OC(O)-aryl, C(O)O-aryl, OC(O)-heteroaryl, C(O)O-heteroaryl, C(O)O-aryl, C(O)O-heteroaryl, C(O)N(R5 )-aryl, C(O)N(R 5 )-heteroaryl, N(R 5 )C(O)-aryl, or N(R 5 )C(O)-heteroaryl, where alkyl, heteroalkyl, aryl, and heteroaryl are each optionally joined by one or more R 9 is replaced by R 9 are each independently C1 to C 20 Alkyl, O-C1-C 20 Alkyl, C1-C 20 heteroalkyl, halo, —CN, OH, oxo, aryl, heteroaryl, O-aryl, or O-heteroaryl), or a pharmaceutically acceptable salt or stereoisomer thereof.

[0010] In some embodiments, B 1 and B 2 each of X is independently a purinyl nucleobase or a pyrimidinyl nucleobase; 1 and X 2 is independently O or S, and Y 1 and Y 2 each independently represents O, S, or NR 5 and L 1 and L 2 each is independently absent, C1-C6 alkyl, or C1-C6 heteroalkyl, where alkyl and heteroalkyl are each optionally selected from R 6 is substituted with R 1 and R 2 each independently represents hydrogen, halo, —CN, C1-C 20 alkyl (e.g., C1-C6 alkyl), or OR 7 and R 3 and R 4 are independently hydrogen, C1 to C 20 Alkyl (e.g., C1-C6 alkyl), C1-C 20heteroalkyl (e.g., C1-C6 heteroalkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, where alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each have 1 to 5 R 8 is substituted with R 5 is hydrogen or C1-C 20 alkyl (e.g., C1-C6 alkyl), and R 6 Halo, -CN, C1~C 20 alkyl (e.g., C1-C6 alkyl), OR 7 , oxo, cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl are each optionally joined by 1 to 5 R 9 is substituted with R 7 is hydrogen, C1 to C 20 alkyl (e.g., C1-C6 alkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, where the alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl are optionally joined by 1 to 5 R 9 is substituted with R 8 are each independently C1 to C 20 alkyl (e.g., C1-C6 alkyl), C(O)-aryl, C(O)-heteroaryl, OC(O)-aryl, C(O)O-aryl, OC(O)-heteroaryl, or C(O)O-heteroaryl, wherein the alkyl, aryl, and heteroaryl are each optionally joined by 1 to 5 R 9 is replaced by R 9 are each independently C1 to C 20 Alkyl, O-C1-C 20 Alkyl, halo, -CN, OH, O-C1~C 20 Alkyl, O-C1-C 20 It is heteroalkyl, O-aryl, or O-heteroaryl.

[0011] In some embodiments, the compound is a compound of formula (Ib), formula (Ic), formula (Id), or formula (Ie) [ka] or a pharmaceutically acceptable salt thereof, wherein B 1 , B 2 , X 1 , X 2 , Y 1 , Y 2 , L 1 , L 2 , R 1 , R 2 , R 3 , R 4 , and each of these subvariables is defined above.

[0012] In some embodiments, B 1 or B 2 At least one of B is a purinyl nucleobase. 1 or B 2 Each of B is independently a purinyl nucleobase. 1 is a purinyl nucleobase. In some embodiments, B 2 is a pyrimidinyl nucleobase. In some embodiments, B 1 is a purinyl nucleobase, and B 2 is a pyrimidinyl nucleobase. In some embodiments, B 1 is adenosinyl or guanosinyl. In some embodiments, B 2 is cytosinyl, thyminyl, or uracilyl. 1 is adenosinyl or guanosinyl, and B 2 is cytosinyl, thyminyl, or uracilyl. 1 Each of B is 2 is independently uracilyl. In some embodiments, B 1 Each of B is 2 are independently adenosinyl.

[0013] In some embodiments, R 1 and R 2 each independently represents hydrogen, halo, or OR 7 In some embodiments, R 1 and R 2 Each of R is independently halo (e.g., fluoro). 1 and R 2 Each of 7 isn't it.

[0014] In some embodiments, X 1 is O. In some embodiments, X 2 is O. In some embodiments, X 1 and X 2 Each of is independently O.

[0015] In some embodiments, Y 1 is O or S. In some embodiments, Y 2 is O or S. In some embodiments, Y 1 and Y 2 Each of Y is independently O or S. In some embodiments, Y 1 or Y 2 One of the two is O and the other is Y 1 or Y 2 and the other is S. In some embodiments, Y 1 or Y 2 Each of Y is independently S. In some embodiments, Y 1 or Y 2 Each of is independently O.

[0016] In some embodiments, L 1 is C1-C6 alkyl (e.g., CH2). In some embodiments, L 2 is C1-C6 alkyl (e.g., CH2). In some embodiments, L 1and L 2 Each of is independently C1-C6 alkyl (eg, CH2).

[0017] In some embodiments, R 3 is hydrogen, aryl, or heteroaryl, where aryl and heteroaryl are optionally joined by 1 to 5 R 8 In some embodiments, R 3 is aryl or heteroaryl, each of which optionally has 1 to 5 R 8 In some embodiments, R 3 is one R 8 is a phenyl substituted with

[0018] In some embodiments, R 4 are independently hydrogen, aryl, or heteroaryl, wherein the aryl and heteroaryl are optionally joined by 1 to 5 R 8 In some embodiments, R 4 is aryl or heteroaryl, each of which optionally has 1 to 5 R 8 In some embodiments, R 4 is one R 8 is a phenyl substituted with

[0019] In some embodiments, R 3 and R 4 is independently hydrogen, aryl, or heteroaryl, wherein the aryl and heteroaryl are optionally joined by 1 to 5 R 8 In some embodiments, R 3 is aryl or heteroaryl, each of which optionally contains 1 to 5 R 8 is substituted with R 4 is hydrogen. In some embodiments, R 3 is one R 8 is phenyl substituted with R 4 is hydrogen. In some embodiments, R3 and R 4 Each of the groups independently represents one R 8 is a phenyl substituted with

[0020] In some embodiments, Y 1 and Y 2 Each of is O and R 3 and R 4 Each of Y is independently hydrogen. 2 is O and R 4 is hydrogen. In some embodiments, Y 1 and Y 2 each independently is S, and R 3 and R 4 Each of the groups independently represents one R 8 In some embodiments, Y 1 is S and R 3 is one R 8 is replaced by .

[0021] In some embodiments, each R 8 are independent, C1~C 20 Alkyl (e.g., C1-C6 alkyl), C1-C 20 Heteroalkyl, C(O)-C1-C 20 Alkyl, OC(O)-C1-C 20 Alkyl, OC(O)O-C1-C 20 Alkyl, OC(O)N(R 5 )-C1~C 20 Alkyl, O-aryl, C(O)-aryl, OC(O)-aryl, or C(O)N(R 5 )-aryl, wherein alkyl, heteroalkyl, aryl, and heteroaryl are each optionally joined by one or more R 9 is replaced by .

[0022] In some embodiments, R 8 is any 1 to 5 R 9 (For example, one R 9 ) is O-C(O)-aryl substituted by

[0023] In some embodiments, R 9 is O-C1~C 12 alkyl (e.g., O-CH(CH)CH). In some embodiments, R 9 is O-C1~C 10 alkyl (e.g., O-CH(CH)CH). In some embodiments, R 9 is O—C1-C8 alkyl (e.g., O—CH2(CH2)6CH3). In some embodiments, R 9 is O-C1-C6 alkyl (for example, O-CH2(CH2)4CH3).

[0024] In some embodiments, the compound is a compound of formula (If) [ka] or a pharmaceutically acceptable salt or stereoisomer thereof, 1 and B 2 each of X is independently a purinyl nucleobase or a pyrimidinyl nucleobase; 1 and X 2 is independently O or S, and Y 1 and Y 2 each independently represents O, S, or NR 5 and L 1 and L 2 Each of C1 to C 20 Alkyl (e.g., C1-C6 alkyl) or C1-C 20 heteroalkyl (e.g., C1-C6 heteroalkyl), where C1-C 20 Alkyl and C1-C 20 Each heteroalkyl is optionally R 6 is replaced by R 1 and R 2 each independently is halo; 3 and R 4 each independently represents hydrogen, C1 to C 20Alkyl (e.g., C1-C6 alkyl), C1-C 20 heteroalkyl (e.g., C1-C6 heteroalkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, where C1-C 20 Alkyl, C1-C 20 Heteroalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each optionally have 1 to 5 R 8 is substituted with R 5 is hydrogen or C1-C6 alkyl, and R 6 Halo, -CN, C1~C 20 alkyl (e.g., C1-C6 alkyl), OR 7 , oxo, cycloalkyl, heterocyclyl, aryl, or heteroaryl, where C1-C 20 Each alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl may optionally have 1 to 5 R 9 is substituted with R 7 is hydrogen, C1 to C 20 alkyl (e.g., C1-C6 alkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, where C1-C 20 Each alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl may optionally have 1 to 5 R 9 is substituted with R 8 are each independently C1 to C 20 alkyl (e.g., C1-C6 alkyl), C(O)-aryl, C(O)-heteroaryl, OC(O)-aryl, or OC(O)-heteroaryl, where C1-C 20 Each of alkyl, C(O)-aryl, C(O)-heteroaryl, OC(O)-aryl, or OC(O)-heteroaryl may optionally be substituted with 1 to 5 R 9 is replaced by R 9 are each independently C1 to C 20 Alkyl, halo, -CN, OH, O-C1~C 20 Alkyl, O-C1-C 20 It is heteroalkyl, O-aryl, or O-heteroaryl.

[0025] In some embodiments, the compound is a compound of formula (Ig) [ka] or a pharmaceutically acceptable salt or stereoisomer thereof, 1 and B 2 each of X is independently a purinyl nucleobase or a pyrimidinyl nucleobase; 1 and X 2 each independently is O; and Y 1 and Y 2 each independently is O or S, and L 1 and L 2 each independently is absent or C1-C6 alkyl; R 1 and R 2 each independently is halo or OH, and R 3 and R 4 each independently represents hydrogen or 1 to 5 R 8 aryl substituted with R 8 each independently represents 1 to 5 R 9 and R is an OC(O)-aryl substituted by 9 are each independently O-C1 to C 12 It is alkyl.

[0026] In some embodiments, the compound of formula (I) is selected from the compounds of Table 1, Table 2, or a pharmaceutically acceptable salt thereof.

[0027] In some embodiments, the compound of Formula (Ia) is selected from the compounds of Table 1, Table 2, or a pharmaceutically acceptable salt thereof.

[0028] In another aspect, the disclosure features a method of treating cancer in a subject, the method includes administering to the subject a compound represented by formula (I): [ka] (wherein Z is either S or O, and B 1 and B 2 each of X is independently a purinyl nucleobase or a pyrimidinyl nucleobase; 1 and X 2 is independently O or S, and Y 1 and Y 2 each independently represents O, S, or NR 5 and L 1 and L 2 each is independently absent, C1-C6 alkyl, or C1-C6 heteroalkyl, where alkyl and heteroalkyl are each optionally selected from R 6 is substituted with R 1 and R 2 each independently represents hydrogen, halo, —CN, C1-C 20 alkyl (e.g., C1-C6 alkyl), or OR 7 and R 3 and R 4 each independently represents hydrogen, C1 to C 20 Alkyl (e.g., C1-C6 alkyl), C1-C 20 Heteroalkyl (e.g., C1-C6 heteroalkyl), OC(O)OC1-C 20 alkyl (e.g., C1-C6 alkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each optionally contain one or more R 8 is substituted with R 5 is hydrogen or C1-C 20 alkyl (e.g., C1-C6 alkyl), and R 6 Halo, -CN, C1~C 20 alkyl (e.g., C1-C6 alkyl), OR 7 , oxo, cycloalkyl, heterocyclyl, aryl, or heteroaryl, where alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl are each independently selected from one or more R 9 and optionally substituted with R 7 is hydrogen, C1 to C20 alkyl (e.g., C1-C6 alkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each optionally contain one or more R 9 is substituted with R 8 are each independently C1 to C 20 Alkyl (e.g., C1-C6 alkyl), C1-C 20 Heteroalkyl, C(O)-C1-C 20 Alkyl, OC(O)-C1-C 20 Alkyl (e.g., C1-C6 alkyl), C(O)O-C1-C 20 Alkyl (e.g., C1-C6 alkyl), OC(O)O-C1-C 20 Alkyl (e.g., C1-C6 alkyl), C(O)N(R 5 )-C1~C 20 Alkyl (e.g., C1-C6 alkyl), N(R 5 )C(O)-C1~C 20 Alkyl (e.g., C1-C6 alkyl), OC(O)N(R 5 )-C1~C 20 Alkyl (e.g., C1-C6 alkyl), O-aryl, O-heteroaryl, C(O)-aryl, C(O)-heteroaryl, OC(O)-aryl, C(O)O-aryl, OC(O)-heteroaryl, C(O)O-heteroaryl, C(O)O-aryl, C(O) O-heteroaryl, C(O)N(R 5 )-aryl, C(O)N(R 5 )-heteroaryl, N(R 5 )C(O)-aryl, N(R 5 )2C(O)-alkyl, or N(R 5 )C(O)-heteroaryl, S(O)N(R 5 )-aryl, wherein alkyl, heteroalkyl, aryl, and heteroaryl are each optionally joined by one or more R 9 is replaced by R 9 are each independently C1 to C 20 Alkyl, O-C1-C 20 Alkyl, C1-C20 heteroalkyl, halo, —CN, OH, oxo, aryl, heteroaryl, O-aryl, or O-heteroaryl), or a pharmaceutically acceptable salt or stereoisomer thereof.

[0029] In some embodiments, the disclosure features a method of treating cancer in a subject, the method includes administering to the subject a compound represented by formula (Ia): [ka] (In the formula, B 1 and B 2 each of X is independently a purinyl nucleobase or a pyrimidinyl nucleobase; 1 and X 2 is independently O or S, and Y 1 and Y 2 each independently represents O, S, or NR 5 and L 1 and L 2 each is independently absent, C1-C6 alkyl, or C1-C6 heteroalkyl, where alkyl and heteroalkyl are each optionally selected from R 6 is substituted with R 1 and R 2 each independently represents hydrogen, halo, —CN, C1-C 20 alkyl (e.g., C1-C6 alkyl), or OR 7 and R 3 and R 4 each independently represents hydrogen, C1 to C 20 Alkyl (e.g., C1-C6 alkyl), C1-C 20 heteroalkyl (e.g., C1-C6 heteroalkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each optionally contain one or more R 8 is substituted with R 5 is hydrogen or C1-C 20 alkyl (e.g., C1-C6 alkyl), and R6 Halo, -CN, C1~C 20 alkyl (e.g., C1-C6 alkyl), OR 7 , oxo, cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl are each optionally joined by one or more R 9 is substituted with R 7 is hydrogen, C1 to C 20 alkyl (e.g., C1-C6 alkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, where alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each optionally contain one or more R 9 is substituted with R 8 are each independently C1 to C 20 Alkyl (e.g., C1-C6 alkyl), C1-C 20 Heteroalkyl, C(O)-C1-C 20 Alkyl, OC(O)-C1-C 20 Alkyl, C(O)O-C1-C 20 Alkyl, OC(O)O-C1-C 20 Alkyl, C(O)N(R 5 )-C1~C 20 Alkyl, N(R 5 )C(O)-C1~C 20 Alkyl, OC(O)N(R 5 )-C1~C 20 Alkyl, O-aryl, O-heteroaryl, C(O)-aryl, C(O)-heteroaryl, OC(O)-aryl, C(O)O-aryl, OC(O)-heteroaryl, C(O)O-heteroaryl, C(O)O-aryl, C(O)O-heteroaryl, C(O)N(R 5 )-aryl, C(O)N(R 5 )-heteroaryl, N(R 5 )C(O)-aryl, or N(R 5 )C(O)-heteroaryl, where alkyl, heteroalkyl, aryl, and heteroaryl are each optionally joined by one or more R 9 is substituted by R 9are each independently C1 to C 20 Alkyl, O-C1-C 20 Alkyl, C1-C 20 heteroalkyl, halo, —CN, OH, oxo, aryl, heteroaryl, O-aryl, or O-heteroaryl), or a pharmaceutically acceptable salt or stereoisomer thereof.

[0030] In some embodiments, the cancer is a cancer of the breast, bone, brain, cervix, colon, gastrointestinal tract, eye, gallbladder, lymph node, blood, lung, liver, skin, oral cavity, prostate, ovary, penis, pancreas, uterus, testicle, stomach, thymus, thyroid, or other body parts (e.g., cancer of the liver). In some embodiments, the cancer differentially expresses STING compared to non-cancerous tissue, such as liver cancer, melanoma, skin cancer, or thyroid cancer.

[0031] In some embodiments, the cancer comprises a PD-1 resistant tumor.

[0032] In some embodiments, the method comprises oral administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the method comprises oral administration of a compound of Formula (Ia) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the method comprises parenteral administration (e.g., subcutaneous, intramuscular, intraperitoneal, or intravenous) of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the method comprises parenteral administration (e.g., subcutaneous, intramuscular, intraperitoneal, or intravenous) of a compound of Formula (Ia) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the method comprises intraperitoneal administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the method comprises intraperitoneal administration of a compound of Formula (Ia) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the method comprises intratumoral administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the method comprises intratumoral administration of a compound of Formula (Ia) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0033] In some embodiments, the method further comprises administering an additional agent (e.g., an anti-cancer or immuno-cancer agent). In some embodiments, the additional agent comprises methotrexate, 5-fluorouracil, doxorubicin, vincristine, bleomycin, vinblastine, dacarbazine, toposide, cisplatin, epirubicin, or sorafenib tosylate.

[0034] In another aspect, the present disclosure provides a vaccine comprising a compound of formula (I) [ka] (wherein Z is either S or O, and B 1 and B 2each of X is independently a purinyl nucleobase or a pyrimidinyl nucleobase; 1 and X 2 is independently O or S, and Y 1 and Y 2 each independently represents O, S, or NR 5 and L 1 and L 2 each is independently absent, C1-C6 alkyl, or C1-C6 heteroalkyl, where alkyl and heteroalkyl are each optionally selected from R 6 is substituted with R 1 and R 2 each independently represents hydrogen, halo, —CN, C1-C 20 alkyl (e.g., C1-C6 alkyl), or OR 7 and R 3 and R 4 Each of C1 to C 20 Alkyl (e.g., C1-C6 alkyl), C1-C 20 Heteroalkyl (e.g., C1-C6 heteroalkyl), OC(O)OC1-C 20 alkyl (e.g., C1-C6 alkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, where alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each optionally contain one or more R 8 is substituted with R 5 is hydrogen or C1-C 20 alkyl (e.g., C1-C6 alkyl), and R 6 Halo, -CN, C1~C 20 alkyl (e.g., C1-C6 alkyl), OR 7 , oxo, cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl are each optionally joined by one or more R 9 is substituted with R 7 is hydrogen, C1 to C 20alkyl (e.g., C1-C6 alkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, where alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each optionally contain one or more R 9 is substituted with R 8 are each independently C1 to C 20 Alkyl (e.g., C1-C6 alkyl), C1-C 20 Heteroalkyl, C(O)-C1-C 20 Alkyl, OC(O)-C1-C 20 Alkyl (e.g., C1-C6 alkyl), C(O)O-C1-C 20 Alkyl (e.g., C1-C6 alkyl), OC(O)O-C1-C 20 Alkyl (e.g., C1-C6 alkyl), C(O)N(R 5 )-C1~C 20 Alkyl (e.g., C1-C6 alkyl), N(R 5 )C(O)-C1~C 20 Alkyl (e.g., C1-C6 alkyl), OC(O)N(R 5 )-C1~C 20 Alkyl (e.g., C1-C6 alkyl), O-aryl, O-heteroaryl, C(O)-aryl, C(O)-heteroaryl, OC(O)-aryl, C(O)O-aryl, OC(O)-heteroaryl, C(O)O-heteroaryl, C(O)O-aryl, C(O)O-heteroaryl, C(O)N(R 5 )-aryl, C(O)N(R 5 )-heteroaryl, N(R 5 )C(O)-aryl, N(R 5 )2C(O)-aryl, or N(R 5 )C(O)-heteroaryl, S(O)N(R 5 )-aryl, wherein alkyl, heteroalkyl, aryl, and heteroaryl are each optionally joined by one or more R 9 is replaced by R 9 are each independently C1 to C 20 Alkyl, O-C1-C 20 Alkyl, C1-C 20and a vaccine adjuvant comprising a compound of the formula (I) or a pharmaceutically acceptable salt or stereoisomer thereof, wherein the compound is a heteroalkyl, halo, -CN, OH, oxo, aryl, heteroaryl, O-aryl, or O-heteroaryl.

[0035] In some embodiments, the present disclosure provides a vaccine comprising a compound of formula (Ia) [ka] or a pharmaceutically acceptable salt or stereoisomer thereof; and a vaccine adjuvant comprising B. 1 and B 2 each of X is independently a purinyl nucleobase or a pyrimidinyl nucleobase; 1 and X 2 each independently is O or S, and Y 1 and Y 2 each independently represents O, S, or NR 5 and L 1 and L 2 each is independently absent, C1-C6 alkyl, or C1-C6 heteroalkyl, where alkyl and heteroalkyl are each optionally selected from R 6 is substituted with R 1 and R 2 each independently represents hydrogen, halo, —CN, C1-C 20 alkyl (e.g., C1-C6 alkyl), or OR 7 and R 3 and R 4 each independently represents hydrogen, C1 to C 20 Alkyl (e.g., C1-C6 alkyl), C1-C 20 heteroalkyl (e.g., C1-C6 heteroalkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each optionally contain one or more R 8 is substituted with R 5 is hydrogen or C1-C 20alkyl (e.g., C1-C6 alkyl), and R 6 Halo, -CN, C1~C 20 alkyl (e.g., C1-C6 alkyl), OR 7 , oxo, cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl are each optionally joined by one or more R 9 is substituted with R 7 is hydrogen, C1 to C 20 alkyl (e.g., C1-C6 alkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each optionally contain one or more R 9 is substituted with R 8 are each independently C1 to C 20 Alkyl (e.g., C1-C6 alkyl), C1-C 20 Heteroalkyl, C(O)-C1-C 20 Alkyl, OC(O)-C1-C 20 Alkyl, C(O)O-C1-C 20 Alkyl, OC(O)O-C1~C 20 Alkyl, C(O)N(R 5 )-C1~C 20 Alkyl, N(R 5 )C(O)-C1~C 20 Alkyl, OC(O)N(R 5 )-C1~C 20 Alkyl, O-aryl, O-heteroaryl, C(O)-aryl, C(O)-heteroaryl, OC(O)-aryl, C(O)O-aryl, OC(O)-heteroaryl, C(O)O-heteroaryl, C(O)O-aryl, C(O)O-heteroaryl, C(O)N(R 5 )-aryl, C(O)N(R 5 )-heteroaryl, N(R 5 )C(O)-aryl, or N(R 5 )C(O)-heteroaryl, where alkyl, heteroalkyl, aryl, and heteroaryl are each optionally joined by one or more R 9is substituted by R 9 are each independently C1 to C 20 Alkyl, O-C1-C 20 Alkyl, C1-C 20 heteroalkyl, halo, -CN, OH, oxo, aryl, heteroaryl, O-aryl, or O-heteroaryl, each of which may optionally be joined by one or more R 10 is substituted with R 10 are each independently C1 to C 20 Alkyl, C1-C 20 Alkenyl, C1-C 20 Alkynyl, C1-C 20 heteroalkyl, halo, -CN, or OH, oxo, and R 5 are each independently hydrogen or C1-C 20 It is alkyl.

[0036] In another aspect, the disclosure features a method of inducing expression of a pattern recognition receptor (PRR) for immune regulation in a subject, the method includes administering to the subject a compound represented by formula (I): [ka] (wherein Z is either S or O, and B 1 and B 2 each of X is independently a purinyl nucleobase or a pyrimidinyl nucleobase; 1 and X 2 each independently is O or S, and Y 1 and Y 2 each independently represents O, S, or NR 5 and L 1 and L 2 each is independently absent, C1-C6 alkyl, or C1-C6 heteroalkyl, where alkyl and heteroalkyl are each optionally selected from R 6 is substituted with R 1 and R 2 each independently represents hydrogen, halo, —CN, C1-C 20alkyl (e.g., C1-C6 alkyl), or OR 7 and R 3 and R 4 each independently represents hydrogen, C1 to C 20 Alkyl (e.g., C1-C6 alkyl), C1-C 20 Heteroalkyl (e.g., C1-C6 heteroalkyl), OC(O)OC1-C 20 alkyl (e.g., C1-C6 alkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each optionally contain one or more R 8 is substituted with R 5 is hydrogen or C1-C 20 alkyl (e.g., C1-C6 alkyl), and R 6 Halo, -CN, C1~C 20 alkyl (e.g., C1-C6 alkyl), OR 7 , oxo, cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl are each optionally joined by one or more R 9 is substituted with R 7 is hydrogen, C1 to C 20 alkyl (e.g., C1-C6 alkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each optionally contain one or more R 9 is substituted with R 8 are each independently C1 to C 20 Alkyl (e.g., C1-C6 alkyl), C1-C 20 Heteroalkyl, C(O)-C1-C 20 Alkyl, OC(O)-C1-C 20 Alkyl (e.g., C1-C6 alkyl), C(O)O-C1-C 20 Alkyl (e.g., C1-C6 alkyl), OC(O)O-C1-C 20 Alkyl (e.g., C1-C6 alkyl), C(O)N(R5 )-C1~C 20 Alkyl (e.g., C1-C6 alkyl), N(R 5 )C(O)-C1~C 20 Alkyl (e.g., C1-C6 alkyl), OC(O)N(R 5 )-C1~C 20 Alkyl (e.g., C1-C6 alkyl), O-aryl, O-heteroaryl, C(O)-aryl, C(O)-heteroaryl, OC(O)-aryl, C(O)O-aryl, OC(O)-heteroaryl, C(O)O-heteroaryl, C(O)O-aryl, C(O)O-heteroaryl, C(O)N(R 5 )-aryl, C(O)N(R 5 )-heteroaryl, N(R 5 )C(O)-aryl, N(R 5 )2C(O)-alkyl, or N(R 5 )C(O)-heteroaryl, S(O)N(R 5 )-aryl, wherein alkyl, heteroalkyl, aryl, and heteroaryl are each optionally joined by one or more R 9 is substituted by R 9 are each independently C1 to C 20 Alkyl, O-C1-C 20 Alkyl, C1-C 20 The present invention includes administering a compound of the formula (I) to a patient in need thereof, the compound comprising: -O-, ...

[0037] In some embodiments, the disclosure features a method of inducing expression of a pattern recognition receptor (PRR) for immunomodulation in a subject, the method comprising administering to the subject a compound of Formula (Ia): [ka] or a pharmaceutically acceptable salt or stereoisomer thereof, 1 and B 2each of X is independently a purinyl nucleobase or a pyrimidinyl nucleobase; 1 and X 2 is independently O or S, and Y 1 and Y 2 each independently represents O, S, or NR 5 and L 1 and L 2 each is independently absent, C1-C6 alkyl, or C1-C6 heteroalkyl, where alkyl and heteroalkyl are each optionally selected from R 6 is substituted with R 1 and R 2 each independently represents hydrogen, halo, —CN, C1-C 20 alkyl (e.g., C1-C6 alkyl), or OR 7 and R 3 and R 4 each independently represents hydrogen, C1 to C 20 Alkyl (e.g., C1-C6 alkyl), C1-C 20 heteroalkyl (e.g., C1-C6 heteroalkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each optionally contain one or more R 8 is substituted with R 5 is hydrogen or C1-C 20 alkyl (e.g., C1-C6 alkyl), and R 6 Halo, -CN, C1~C 20 alkyl (e.g., C1-C6 alkyl), OR 7 , oxo, cycloalkyl, heterocyclyl, aryl, or heteroaryl, where alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl are each independently selected from one or more R 9 and optionally substituted with R 7 is hydrogen, C1 to C 20alkyl (e.g., C1-C6 alkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each optionally contain one or more R 9 is substituted with R 8 are each independently C1 to C 20 Alkyl (e.g., C1-C6 alkyl), C1-C 20 Heteroalkyl, C(O)-C1-C 20 Alkyl, OC(O)-C1-C 20 Alkyl, C(O)O-C1-C 20 Alkyl, OC(O)O-C1~C 20 Alkyl, C(O)N(R 5 )-C1~C 20 Alkyl, N(R 5 )C(O)-C1~C 20 Alkyl, OC(O)N(R 5 )-C1~C 20 Alkyl, O-aryl, O-heteroaryl, C(O)-aryl, C(O)-heteroaryl, OC(O)-aryl, C(O)O-aryl, OC(O)-heteroaryl, C(O)O-heteroaryl, C(O)O-aryl, C(O)O-heteroaryl, C(O)N(R 5 )-aryl, C(O)N(R 5 )-heteroaryl, N(R 5 )C(O)-aryl, or N(R 5 )C(O)-heteroaryl, where alkyl, heteroalkyl, aryl, and heteroaryl are each optionally joined by one or more R 9 is substituted by R 9 are each independently C1 to C 20 Alkyl, O-C1-C 20 Alkyl, C1-C 20 It is heteroalkyl, halo, -CN, OH, oxo, aryl, heteroaryl, O-aryl, or O-heteroaryl.

[0038] In another aspect, the disclosure features a method of inducing expression of a pattern recognition receptor (PRR) for immune regulation and inducing a therapeutic response in a subject suffering from cancer, the method comprising administering to the subject a compound represented by formula (I): [ka] (wherein Z is either S or O, and B 1 and B 2 each of X is independently a purinyl nucleobase or a pyrimidinyl nucleobase; 1 and X 2 is independently O or S, and Y 1 and Y 2 each independently represents O, S, or NR 5 and L 1 and L 2 each independently is absent, C1-C6 alkyl, or C1-C6 heteroalkyl, wherein each alkyl and heteroalkyl is optionally joined to R 6 is substituted with R 1 and R 2 each independently represents hydrogen, halo, —CN, C1-C 20 alkyl (e.g., C1-C6 alkyl), or OR 7 and R 3 and R 4 each independently represents hydrogen, C1 to C 20 Alkyl (e.g., C1-C6 alkyl), C1-C 20 Heteroalkyl (e.g., C1-C6 heteroalkyl), OC(O)OC1-C 20 alkyl (e.g., C1-C6 alkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each optionally contain one or more R 8 is substituted with R 5 is hydrogen or C1-C 20 alkyl (e.g., C1-C6 alkyl), and R 6 Halo, -CN, C1~C 20 alkyl (e.g., C1-C6 alkyl), OR7 , oxo, cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl are each optionally joined by one or more R 9 is substituted with R 7 is hydrogen, C1 to C 20 alkyl (e.g., C1-C6 alkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each optionally contain one or more R 9 is substituted with R 8 are each independently C1 to C 20 Alkyl (e.g., C1-C6 alkyl), C1-C 20 Heteroalkyl, C(O)-C1-C 20 Alkyl, OC(O)-C1-C 20 Alkyl (e.g., C1-C6 alkyl), C(O)O-C1-C 20 Alkyl (e.g., C1-C6 alkyl), OC(O)O-C1-C 20 Alkyl (e.g., C1-C6 alkyl), C(O)N(R 5 )-C1~C 20 Alkyl (e.g., C1-C6 alkyl), N(R 5 )C(O)-C1~C 20 Alkyl (e.g., C1-C6 alkyl), OC(O)N(R 5 )-C1~C 20 Alkyl (e.g., C1-C6 alkyl), O-aryl, O-heteroaryl, C(O)-aryl, C(O)-heteroaryl, OC(O)-aryl, C(O)O-aryl, OC(O)-heteroaryl, C(O)O-heteroaryl, C(O)O-aryl, C(O)O-heteroaryl, C(O)N(R 5 )-aryl, C(O)N(R 5 )-heteroaryl, N(R 5 )C(O)-aryl, N(R 5 )2C(O)-aryl, or N(R 5 )C(O)-heteroaryl, S(O)N(R 5)-aryl, wherein alkyl, heteroalkyl, aryl, and heteroaryl are each optionally joined by one or more R 9 is substituted by R 9 are each independently C1 to C 20 Alkyl, O-C1-C 20 Alkyl, C1-C 20 Heteroalkyl, O-C1-C 20 -NR 10 R 10 , halo, —CN, OH, oxo, aryl, heteroaryl, O-aryl, or O-heteroaryl), or a pharmaceutically acceptable salt or stereoisomer thereof.

[0039] In some embodiments, the disclosure features a method of inducing expression of a pattern recognition receptor (PRR) for immune regulation and inducing a therapeutic response in a subject suffering from cancer, the method comprising administering to the subject a compound of Formula (Ia): [ka] or a pharmaceutically acceptable salt or stereoisomer thereof, 1 and B 2 each of X is independently a purinyl nucleobase or a pyrimidinyl nucleobase; 1 and X 2 is independently O or S, and Y 1 and Y 2 each independently represents O, S, or NR 5 and L 1 and L 2 each independently is absent, C1-C6 alkyl, or C1-C6 heteroalkyl, wherein each alkyl and heteroalkyl is optionally joined to R 6 is substituted with R 1 and R 2 each independently represents hydrogen, halo, —CN, C1-C 20 alkyl (e.g., C1-C6 alkyl), or OR 7 and R 3 and R 4each independently represents hydrogen, C1 to C 20 Alkyl (e.g., C1-C6 alkyl), C1-C 20 heteroalkyl (e.g., C1-C6 heteroalkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each optionally contain one or more R 8 is substituted with R 5 is hydrogen or C1-C 20 alkyl (e.g., C1-C6 alkyl), and R 6 Halo, -CN, C1~C 20 alkyl (e.g., C1-C6 alkyl), OR 7 , oxo, cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl are each optionally joined by one or more R 9 is substituted with R 7 is hydrogen, C1 to C 20 alkyl (e.g., C1-C6 alkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each optionally contain one or more R 9 is substituted with R 8 are each independently C1 to C 20 Alkyl (e.g., C1-C6 alkyl), C1-C 20 Heteroalkyl, C(O)-C1-C 20 Alkyl, OC(O)-C1-C 20 Alkyl, C(O)O-C1-C 20 Alkyl, OC(O)O-C1-C 20 Alkyl, C(O)N(R 5 )-C1~C 20 Alkyl, N(R 5 )C(O)-C1~C 20 Alkyl, OC(O)N(R 5 )-C1~C 20Alkyl, O-aryl, O-heteroaryl, C(O)-aryl, C(O)-heteroaryl, OC(O)-aryl, C(O)O-aryl, OC(O)-heteroaryl, C(O)O-heteroaryl, C(O)O-aryl, C(O)O-heteroaryl, C(O)N(R 5 )-aryl, C(O)N(R 5 )-heteroaryl, N(R 5 )C(O)-aryl, or N(R 5 )C(O)-heteroaryl, where alkyl, heteroalkyl, aryl, and heteroaryl are each optionally joined by one or more R 9 is substituted by R 9 are each independently C1 to C 20 Alkyl, O-C1-C 20 Alkyl, C1-C 20 It is heteroalkyl, halo, -CN, OH, oxo, aryl, heteroaryl, O-aryl, or O-heteroaryl.

[0040] In another aspect, the disclosure features a method of inducing an immune response in a subject, the method includes administering to the subject a compound represented by formula (I): [ka] (wherein Z is either S or O, and B 1 and B 2 each of X is independently a purinyl nucleobase or a pyrimidinyl nucleobase; 1 and X 2 each independently is O or S; and Y 1 and Y 2 each independently represents O, S, or NR 5 and L 1 and L 2 each independently is absent, C1-C6 alkyl, or C1-C6 heteroalkyl, wherein each alkyl and heteroalkyl is optionally joined to R 6 is substituted with R 1 and R 2 each independently represents hydrogen, halo, —CN, C1-C 20alkyl (e.g., C1-C6 alkyl), or OR 7 and R 3 and R 4 each independently represents hydrogen, C1 to C 20 Alkyl (e.g., C1-C6 alkyl), C1-C 20 Heteroalkyl (e.g., C1-C6 heteroalkyl), OC(O)OC1-C 20 alkyl (e.g., C1-C6 alkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each optionally contain one or more R 8 is substituted with R 5 is hydrogen or C1-C 20 alkyl (e.g., C1-C6 alkyl), and R 6 Halo, -CN, C1~C 20 alkyl (e.g., C1-C6 alkyl), OR 7 , oxo, cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl are each optionally joined by one or more R 9 is substituted with R 7 is hydrogen, C1 to C 20 alkyl (e.g., C1-C6 alkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl are each independently optionally represented by one or more R 9 is substituted with R 8 are each independently C1 to C 20 Alkyl (e.g., C1-C6 alkyl), C1-C 20 Heteroalkyl, C(O)-C1-C 20 Alkyl, OC(O)-C1-C 20 Alkyl (e.g., C1-C6 alkyl), C(O)O-C1-C 20 Alkyl (e.g., C1-C6 alkyl), OC(O)O-C1-C 20 Alkyl (e.g., C1-C6 alkyl), C(O)N(R5 )-C1~C 20 Alkyl (e.g., C1-C6 alkyl), N(R 5 )C(O)-C1~C 20 Alkyl (e.g., C1-C6 alkyl), OC(O)N(R 5 )-C1~C 20 Alkyl (e.g., C1-C6 alkyl), O-aryl, O-heteroaryl, C(O)-aryl, C(O)-heteroaryl, OC(O)-aryl, C(O)O-aryl, OC(O)-heteroaryl, C(O)O-heteroaryl, C(O)O-aryl, C(O)O-heteroaryl, C(O)N(R 5 )-aryl, C(O)N(R 5 )-heteroaryl, N(R 5 )C(O)-aryl, N(R 5 )2C(O)-alkyl, or N(R 5 )C(O)-heteroaryl, S(O)N(R 5 )-aryl, wherein alkyl, heteroalkyl, aryl, and heteroaryl are each independently optionally joined by one or more R 9 is substituted by R 9 are each independently C1 to C 20 Alkyl, O-C1-C 20 Alkyl, C1-C 20 heteroalkyl, halo, —CN, OH, oxo, aryl, heteroaryl, O-aryl, or O-heteroaryl), or a pharmaceutically acceptable salt or stereoisomer thereof.

[0041] In some embodiments, the disclosure features a method of inducing an immune response in a subject, the method comprising administering to the subject a compound of formula (Ia): [ka] (In the formula, B 1 and B 2 each of X is independently a purinyl nucleobase or a pyrimidinyl nucleobase; 1 and X 2 each independently is O or S; and Y1 and Y 2 each independently represents O, S, or NR 5 and L 1 and L 2 each independently is absent, C1-C6 alkyl, or C1-C6 heteroalkyl, wherein each alkyl and heteroalkyl is optionally joined to R 6 is substituted with R 1 and R 2 each independently represents hydrogen, halo, —CN, C1-C 20 alkyl (e.g., C1-C6 alkyl), or OR 7 and R 3 and R 4 are independently hydrogen, C1 to C 20 Alkyl (e.g., C1-C6 alkyl), C1-C 20 heteroalkyl (e.g., C1-C6 heteroalkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each optionally contain one or more R 8 is substituted with R 5 is hydrogen or C1-C 20 alkyl (e.g., C1-C6 alkyl), and R 6 Halo, -CN, C1~C 20 alkyl (e.g., C1-C6 alkyl), OR 7 , oxo, cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl are each optionally joined by one or more R 9 is substituted with R 7 is hydrogen, C1 to C 20 alkyl (e.g., C1-C6 alkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each optionally contain one or more R 9 is substituted with R 8 are each independently C1 to C 20Alkyl (e.g., C1-C6 alkyl), C1-C 20 Heteroalkyl, C(O)-C1-C 20 Alkyl, OC(O)-C1-C 20 Alkyl, C(O)O-C1-C 20 Alkyl, OC(O)O-C1~C 20 Alkyl, C(O)N(R 5 )-C1~C 20 Alkyl, N(R 5 )C(O)-C1~C 20 Alkyl, OC(O)N(R 5 )-C1~C 20 Alkyl, O-aryl, O-heteroaryl, C(O)-aryl, C(O)-heteroaryl, OC(O)-aryl, C(O)O-aryl, OC(O)-heteroaryl, C(O)O-heteroaryl, C(O)O-aryl, C(O)O-heteroaryl, C(O)N(R 5 )-aryl, C(O)N(R 5 )-heteroaryl, N(R 5 )C(O)-aryl, or N(R 5 )C(O)-heteroaryl, where alkyl, heteroalkyl, aryl, and heteroaryl are each optionally joined by one or more R 9 is substituted by R 9 are each independently C1 to C 20 Alkyl, O-C1-C 20 Alkyl, C1-C 20 heteroalkyl, halo, —CN, OH, oxo, aryl, heteroaryl, O-aryl, or O-heteroaryl), or a pharmaceutically acceptable salt or stereoisomer thereof.

[0042] In some embodiments, the immune response comprises anti-tumor immunity. In some embodiments, the immune response comprises induction of a PRR (e.g., STING, RIG-I, MDA5). [Brief explanation of the drawings]

[0043] [Figure 1A]Exemplary compounds are shown to bind to STING to activate type I IFN signaling. Figure 1A illustrates the results of a primary STING agonist screen, in which a compound library was screened using HEK293 cells stably expressing the ISG54 (ISRE)-promoter-driven firefly luciferase gene. Cells transfected with human STING and the internal control Renilla luciferase were treated with 25 μM exemplary compounds, and IRF activity was assessed by measuring luciferase levels. [Figure 1B] Exemplary compounds are shown to bind to STING to activate type I IFN signaling. Figure 1A illustrates the results of a primary STING agonist screen, in which a compound library was screened using HEK293 cells stably expressing the ISG54 (ISRE)-promoter-driven firefly luciferase gene. Cells transfected with human STING and the internal control Renilla luciferase were treated with 25 μM exemplary compounds, and IRF activity was assessed by measuring luciferase levels. [Figure 1C] Exemplary compounds are shown to bind to STING to activate type I IFN signaling. Figure 1A illustrates the results of a primary STING agonist screen, in which a compound library was screened using HEK293 cells stably expressing the ISG54 (ISRE)-promoter-driven firefly luciferase gene. Cells transfected with human STING and the internal control Renilla luciferase were treated with 25 μM exemplary compounds, and IRF activity was assessed by measuring luciferase levels. [Figure 2A] 1 shows a potency comparison of an exemplary compound (Compound 1) with the natural STING ligand, 2′-3′cGAMP. [Figure 2B] 1 shows a potency comparison of an exemplary compound (Compound 1) with the natural STING ligand, 2′-3′cGAMP. [Figure 2C] 1 shows a potency comparison of an exemplary compound (Compound 1) with the natural STING ligand, 2′-3′cGAMP. [Figure 2D] 1 shows a potency comparison of an exemplary compound (Compound 1) with the natural STING ligand, 2′-3′cGAMP. [Figure 2E] 1 shows a potency comparison of an exemplary compound (Compound 1) with the natural STING ligand, 2′-3′cGAMP. [Figure 2F] 1 shows a potency comparison of an exemplary compound (Compound 1) with the natural STING ligand, 2′-3′cGAMP. [Figure 3A] 1 shows that exemplary compounds have STING-dependent activity. [Figure 3B] 1 shows that exemplary compounds have STING-dependent activity. [Figure 4] 1 shows induction of IRFs by exemplary compounds. [Figure 5A] 1 shows that exemplary compounds bind to STING and activate STING-dependent type I IFN and NF-κB signaling in HEK293 cells. [Figure 5B] 1 shows that exemplary compounds bind to STING and activate STING-dependent type I IFN and NF-κB signaling in HEK293 cells. [Figure 6] 1 shows the induction of NF-κB by exemplary compounds. [Figure 7A] Exemplary compounds are shown to induce apoptotic cell death through modulation of BAX and BCL-2 levels. [Figure 7B] Exemplary compounds are shown to induce apoptotic cell death through modulation of BAX and BCL-2 levels. [Figure 7C] Exemplary compounds are shown to induce apoptotic cell death through modulation of BAX and BCL-2 levels. [Figure 7D] Exemplary compounds are shown to induce apoptotic cell death through modulation of BAX and BCL-2 levels. [Figure 7E] Exemplary compounds are shown to induce apoptotic cell death through modulation of BAX and BCL-2 levels. [Figure 8A] 1 shows selective induction of apoptosis by Compound 1 in acute monocytic leukemia cell line (THP1) and PBMCs. [Figure 8B] 1 shows selective induction of apoptosis by Compound 1 in acute monocytic leukemia cell line (THP1) and PBMCs. [Figure 9A] We show that an exemplary compound (Compound 1) induces selective and enhanced induction of ISG- and PRR-related genes in an acute monocytic leukemia cell line (THP1) compared to primary PBMCs. Gene expression analysis was performed in THP1 and PBMCs. [Figure 9B] We show that an exemplary compound (Compound 1) induces selective and enhanced induction of ISG- and PRR-related genes in an acute monocytic leukemia cell line (THP1) compared to primary PBMCs. Gene expression analysis was performed in THP1 and PBMCs. [Figure 10A] Exemplary compounds are shown to inhibit tumor cell growth. [Figure 10B] Exemplary compounds are shown to inhibit tumor cell growth. [Figure 11A] Exemplary compounds are shown to have STING-dependent IRF activity but do not cause induction of NF-κB. [Figure 11B] Exemplary compounds are shown to have STING-dependent IRF activity but do not cause induction of NF-κB. [Figure 12] 1 shows that exemplary compounds activate IRF signaling in THP1 cells. [Figure 13A] Exemplary compounds are shown to have activity similar to the natural STING ligand, 2'-3'cGAMP. [Figure 13B] Exemplary compounds are shown to have activity similar to the natural STING ligand, 2'-3'cGAMP. [Figure 13C] Exemplary compounds are shown to have activity similar to the natural STING ligand, 2'-3'cGAMP. [Figure 13D] Exemplary compounds are shown to have activity similar to the natural STING ligand, 2'-3'cGAMP. [Figure 14] Exemplary compounds are shown to bind directly to STING. [Figure 15A] 1 shows that exemplary compounds have STING-dependent IRF activity but do not cause induction of NF-κB. [Figure 15B] 1 shows that exemplary compounds have STING-dependent IRF activity but do not cause induction of NF-κB. [Figure 16] 1 shows that exemplary compounds bind directly to STING. [Figure 17A] 1 shows that exemplary compounds have STING-dependent activity. [Figure 17B] 1 shows that exemplary compounds have STING-dependent activity. [Figure 18A] 1 shows that exemplary compounds have similar potency to the natural STING ligand, 2′-3′cGAMP. [Figure 18B] 1 shows that exemplary compounds have similar potency to the natural STING ligand, 2′-3′cGAMP. [Figure 18C] 1 shows that exemplary compounds have similar potency to the natural STING ligand, 2′-3′cGAMP. [Figure 18D] 1 shows that exemplary compounds have similar potency to the natural STING ligand, 2′-3′cGAMP. [Figure 19A] 1 shows that exemplary compounds have enhanced activity in an acute monocytic leukemia cell line (THP1) compared to primary cell PBMC. [Figure 19B] 1 shows that exemplary compounds have enhanced activity in an acute monocytic leukemia cell line (THP1) compared to primary cell PBMC. [Figure 20A]1 shows IRF induction by exemplary compounds. [Figure 20B] 1 shows IRF induction by exemplary compounds. [Figure 20C] 1 shows IRF induction by exemplary compounds. [Figure 21A] 1 shows IRF induction by exemplary compounds. [Figure 21B] 1 shows IRF induction by exemplary compounds. [Figure 21C] 1 shows IRF induction by exemplary compounds. [Figure 22A] 22A and 22B are graphs showing evaluation of the % IRF induction (Figure 22A) and % NF-κB induction (Figure 22B) by Compound 1, Compound 1A, and Compound 1B in THP1 dual cells carrying both a secreted embryonic alkaline phosphatase (SEAP) reporter gene under the control of an IFN-β minimal promoter fused to five copies of the NF-κB consensus transcription response element and a Lucia reporter gene under the control of an ISG54 minimal promoter. [Figure 22B] 22A and 22B are graphs showing evaluation of the % IRF induction (Figure 22A) and % NF-κB induction (Figure 22B) by Compound 1, Compound 1A, and Compound 1B in THP1 dual cells carrying both a secreted embryonic alkaline phosphatase (SEAP) reporter gene under the control of an IFN-β minimal promoter fused to five copies of the NF-κB consensus transcription response element and a Lucia reporter gene under the control of an ISG54 minimal promoter. [Figure 23A] 23A-23D are graphs showing induction of IRF (FIGS. 23A-23B) and NF-κB (FIGS. 23C-23D) by Compound 1, demonstrating that Compound 1 was taken up by cells without the use of transfection agents. [Figure 23B] 23A-23D are graphs showing induction of IRF (FIGS. 23A-23B) and NF-κB (FIGS. 23C-23D) by Compound 1, demonstrating that Compound 1 was taken up by cells without the use of transfection agents. [Figure 23C]23A-23D are graphs showing induction of IRF (FIGS. 23A-23B) and NF-κB (FIGS. 23C-23D) by Compound 1, demonstrating that Compound 1 was taken up by cells without the use of transfection agents. [Figure 23D] 23A-23D are graphs showing induction of IRF (FIGS. 23A-23B) and NF-κB (FIGS. 23C-23D) by Compound 1, demonstrating that Compound 1 was taken up by cells without the use of transfection agents. [Figure 24A] 1 is a graph showing the induction of IRF by Compound 3, demonstrating that Compound 3 was taken up into cells without the use of a transfection agent. [Figure 24B] 1 is a graph showing the induction of IRF by Compound 3, demonstrating that Compound 3 was taken up into cells without the use of a transfection agent. [Figure 25A] 25A-25D are graphs showing the induction of IRF (FIGS. 25A-25B) and NF-κB (FIGS. 25C-25D) by compound 12, demonstrating that compound 12 was taken up by cells without the use of transfection agents. [Figure 25B] 25A-25D are graphs showing the induction of IRF (FIGS. 25A-25B) and NF-κB (FIGS. 25C-25D) by compound 12, demonstrating that compound 12 was taken up by cells without the use of transfection agents. [Figure 25C] 25A-25D are graphs showing the induction of IRF (FIGS. 25A-25B) and NF-κB (FIGS. 25C-25D) by compound 12, demonstrating that compound 12 was taken up by cells without the use of transfection agents. [Figure 25D] 25A-25D are graphs showing the induction of IRF (FIGS. 25A-25B) and NF-κB (FIGS. 25C-25D) by compound 12, demonstrating that compound 12 was taken up by cells without the use of transfection agents. [Figure 26A] 26A-26D are graphs showing the induction of IRF (FIGS. 26A-26B) and NF-κB (FIGS. 26C-26D) by compound 13, demonstrating that compound 13 was taken up by cells without the use of transfection agents. [Figure 26B]26A-26D are graphs showing the induction of IRF (FIGS. 26A-26B) and NF-κB (FIGS. 26C-26D) by compound 13, demonstrating that compound 13 was taken up by cells without the use of transfection agents. [Figure 26C] 26A-26D are graphs showing the induction of IRF (FIGS. 26A-26B) and NF-κB (FIGS. 26C-26D) by compound 13, demonstrating that compound 13 was taken up by cells without the use of transfection agents. [Figure 26D] 26A-26D are graphs showing the induction of IRF (FIGS. 26A-26B) and NF-κB (FIGS. 26C-26D) by compound 13, demonstrating that compound 13 was taken up by cells without the use of transfection agents. [Figure 27A] 27A-27D are graphs showing the induction of IRF (FIGS. 27A-27B) and NF-κB (FIGS. 27C-27D) by compound 14, demonstrating that compound 14 was taken up by cells without the use of transfection agents. [Figure 27B] 27A-27D are graphs showing the induction of IRF (FIGS. 27A-27B) and NF-κB (FIGS. 27C-27D) by compound 14, demonstrating that compound 14 was taken up by cells without the use of transfection agents. [Figure 27C] 27A-27D are graphs showing the induction of IRF (FIGS. 27A-27B) and NF-κB (FIGS. 27C-27D) by compound 14, demonstrating that compound 14 was taken up by cells without the use of transfection agents. [Figure 27D] 27A-27D are graphs showing the induction of IRF (FIGS. 27A-27B) and NF-κB (FIGS. 27C-27D) by compound 14, demonstrating that compound 14 was taken up by cells without the use of transfection agents. [Figure 28A] 28A-28D are graphs showing the induction of IRF (FIGS. 28A-28B) and NF-κB (FIGS. 28C-28D) by compound 15, demonstrating that compound 15 was taken up by cells without the use of transfection agents. [Figure 28B]28A-28D are graphs showing the induction of IRF (FIGS. 28A-28B) and NF-κB (FIGS. 28C-28D) by compound 15, demonstrating that compound 15 was taken up by cells without the use of transfection agents. [Figure 28C] 28A-28D are graphs showing the induction of IRF (FIGS. 28A-28B) and NF-κB (FIGS. 28C-28D) by compound 15, demonstrating that compound 15 was taken up by cells without the use of transfection agents. [Figure 28D] 28A-28D are graphs showing the induction of IRF (FIGS. 28A-28B) and NF-κB (FIGS. 28C-28D) by compound 15, demonstrating that compound 15 was taken up by cells without the use of transfection agents. [Figure 29A] FIG. 29A compares the induction of IRF (FIG. 29A) and NF-κB (FIG. 29B) by Compound 1, Compound 3, Compound 12, Compound 13, Compound 14, and Compound 15. [Figure 29B] FIG. 29A compares the induction of IRF (FIG. 29A) and NF-κB (FIG. 29B) by Compound 1, Compound 3, Compound 12, Compound 13, Compound 14, and Compound 15. [Figure 30A] 30A and 30B are graphs showing the stability of Compound 1 in serum (FIG. 30A) and microsomes (FIG. 30B). In FIG. 30B, Peak 1 and Peak 2 represent Compound 1-A and Compound 1-B, respectively. [Figure 30B] 30A and 30B are graphs showing the stability of Compound 1 in serum (FIG. 30A) and microsomes (FIG. 30B). In FIG. 30B, Peak 1 and Peak 2 represent Compound 1-A and Compound 1-B, respectively. [Figure 31A] 31A and 31B are graphs showing the stability of compound 15 in serum (FIG. 31A) and microsomes (FIG. 31B). In FIG. 31B, peak 1 and peak 2 represent compound 15-A and compound 15-B, respectively. [Figure 31B] 31A and 31B are graphs showing the stability of compound 15 in serum (FIG. 31A) and microsomes (FIG. 31B). In FIG. 31B, peak 1 and peak 2 represent compound 15-A and compound 15-B, respectively. [Figure 32A] FIG. 32A compares the induction of IRF (FIG. 32A) and NF-κB (FIG. 32B) by compound 15 and its isomers, compound 15-A and compound 15-B. [Figure 32B] FIG. 32A compares the induction of IRF (FIG. 32A) and NF-κB (FIG. 32B) by compound 15 and its isomers, compound 15-A and compound 15-B. [Figure 33] FIG. 1 shows the induction of apoptosis through the cytotoxicity rate of THP1 cells by Compound 15 and its isomers Compound 15-A and Compound 15-B. [Figure 34A] Similar to 2',3'-cGAMP, we show that compound 1 binds to STING and activates type 1 IFN signaling. [Figure 34B] Similar to 2',3'-cGAMP, we show that compound 1 binds to STING and activates type 1 IFN signaling. [Figure 35] FIG. 1 shows that, like 2′,3′-cGAMP, compound 1 is highly active in activating type 1 IFN signaling in mouse macrophages. [Figure 36A] 36A and 36B are graphs showing that Compound 1, Compound 5, Compound 12, Compound 13, Compound 14, and Compound 15 are more active against the natural STING ligand 3',3'-cGAMP in human monocytes (FIG. 36A) and mouse macrophages (FIG. 36B). [Figure 36B] 36A and 36B are graphs showing that Compound 1, Compound 5, Compound 12, Compound 13, Compound 14, and Compound 15 are more active against the natural STING ligand 3',3'-cGAMP in human monocytes (FIG. 36A) and mouse macrophages (FIG. 36B). [Figure 37A] 37A and 37B are graphs showing induction of type I IFN signaling in HEK293 cells (FIG. 37A) and THP1 cells (FIG. 37B) by Compound 1 and its isomers Compound 1A and Compound 1B. [Figure 37B]37A and 37B are graphs showing induction of type I IFN signaling in HEK293 cells (FIG. 37A) and THP1 cells (FIG. 37B) by Compound 1 and its isomers Compound 1A and Compound 1B. [Figure 38A] 38A and 38B show that compound 1 and compound 15 induce the production of type III interferon (IL-29) in THP1 cells (FIG. 38A), and that both compound 1 and compound 15 are taken up by the cells without the use of transfection reagents (FIG. 38B). [Figure 38B] 38A and 38B show that compound 1 and compound 15 induce the production of type III interferon (IL-29) in THP1 cells (FIG. 38A), and that both compound 1 and compound 15 are taken up by the cells without the use of transfection reagents (FIG. 38B). [Figure 39A] 1 is a graph comparing the induction of type I IFN signaling in THP1 cells by Compound 1, Compound 13, and Compound 15 as STING agonists. [Figure 39B] 1 is a graph comparing the induction of type I IFN signaling in THP1 cells by Compound 1, Compound 13, and Compound 15 as STING agonists. [Figure 40A] FIG. 42A compares the induction of IRF (FIG. 42A) and NF-κB (FIG. 42B) by compound 15 and compound 16. [Figure 40B] FIG. 42A compares the induction of IRF (FIG. 42A) and NF-κB (FIG. 42B) by compound 15 and compound 16. [Figure 41A] We show that Compound 1 can activate the major human STING-HAQ polymorphic variant. [Figure 41B] We show that Compound 1 can activate the major human STING-HAQ polymorphic variant. [Figure 42]We show that residues R238 and Y167 of loss-of-function STING mutants generated in our laboratory (STING-R238A and STING-Y167A) are important for cGAMP activation of Compound 1 and the STING-dependent IFN response. [Figure 43] 1 shows IRF-type I IFN activity by Compound 1 in a co-cultured tumor / THP1 cell line. [Figure 44A] We demonstrate that compound 1 inhibits tumor cell growth in tumor cells and THP1 cells using a rich image-based approach and that compound 1 is STING-dependent. [Figure 44B] We demonstrate that compound 1 inhibits tumor cell growth in tumor cells and THP1 cells using a rich image-based approach and that compound 1 is STING-dependent. [Figure 45A] Figure 1 shows that Compound 1 induces apoptosis in acute monocytic leukemia cells. [Figure 45B] Figure 1 shows that Compound 1 induces apoptosis in acute monocytic leukemia cells. [Figure 46A] 1 shows that compound 1 induces apoptosis in the mouse lymphoma cell line A20. [Figure 46B] 1 shows that compound 1 induces apoptosis in the mouse lymphoma cell line A20. [Figure 46C] 1 shows that compound 1 induces apoptosis in the mouse lymphoma cell line A20. [Figure 46D] 1 shows that compound 1 induces apoptosis in the mouse lymphoma cell line A20. [Figure 46E] 1 shows that compound 1 induces apoptosis in the mouse lymphoma cell line A20. [Figure 47A] 1 shows that Compound 1 induces apoptosis in mouse melanoma cells. [Figure 47B] 1 shows that Compound 1 induces apoptosis in mouse melanoma cells. [Figure 48A]Compound 1 inhibits mouse A20 B-cell lymphoma tumor cells. [Figure 48B] Compound 1 inhibits mouse A20 B-cell lymphoma tumor cells. [Figure 48C] Compound 1 inhibits mouse A20 B-cell lymphoma tumor cells. [Figure 48D] Compound 1 inhibits mouse A20 B-cell lymphoma tumor cells. [Figure 49] 1 shows the antitumor activity of compound 1 using an information-rich image-based approach. [Figure 50A] This shows that cell death induction by Compound 1 is mediated by STING. [Figure 50B] This shows that cell death induction by Compound 1 is mediated by STING. [Figure 50C] This shows that cell death induction by Compound 1 is mediated by STING. [Figure 50D] This shows that cell death induction by Compound 1 is mediated by STING. [Figure 50E] This shows that cell death induction by Compound 1 is mediated by STING. [Figure 50F] This shows that cell death induction by Compound 1 is mediated by STING. [Figure 51A] Figure 51 shows the results of a gel shift assay demonstrating that Compound 1 binds to STING. Close structural analogs of Compound 1 bearing fluorescent substituents were synthesized for gel shift assays. Figure 51A shows 250 μM Compound 1 analog with 20 μM to 0 μM STING. Figure 51B shows 10 μM STING with 1 mM to 0 mM Compound 1 analog. Figure 51C shows an immunoblot for detecting STING. [Figure 51B]Figure 51 shows the results of a gel shift assay demonstrating that Compound 1 binds to STING. Close structural analogs of Compound 1 bearing fluorescent substituents were synthesized for gel shift assays. Figure 51A shows 250 μM Compound 1 analog with 20 μM to 0 μM STING. Figure 51B shows 10 μM STING with 1 mM to 0 mM Compound 1 analog. Figure 51C shows an immunoblot for detecting STING. [Figure 51C] Figure 51 shows the results of a gel shift assay demonstrating that Compound 1 binds to STING. Close structural analogs of Compound 1 bearing fluorescent substituents were synthesized for gel shift assays. Figure 51A shows 250 μM Compound 1 analog with 20 μM to 0 μM STING. Figure 51B shows 10 μM STING with 1 mM to 0 mM Compound 1 analog. Figure 51C shows an immunoblot for detecting STING. [Figure 52A] 52A-52P show analysis of IRF3 and NF-kB pathways after treatment with Compound 1. Figures 52A-52P show immunoblots of THP-1 cells treated with 5 μM Compound 1 or 2′-3′cGAMP. [Figure 52B] 52A-52P show analysis of IRF3 and NF-kB pathways after treatment with Compound 1. Figures 52A-52P show immunoblots of THP-1 cells treated with 5 μM Compound 1 or 2′-3′cGAMP. [Figure 52C] 52A-52P show analysis of IRF3 and NF-kB pathways after treatment with Compound 1. Figures 52A-52P show immunoblots of THP-1 cells treated with 5 μM Compound 1 or 2′-3′cGAMP. [Figure 52D] 52A-52P show analysis of IRF3 and NF-kB pathways after treatment with Compound 1. Figures 52A-52P show immunoblots of THP-1 cells treated with 5 μM Compound 1 or 2′-3′cGAMP. [Figure 52E] 52A-52P show analysis of IRF3 and NF-kB pathways after treatment with Compound 1. Figures 52A-52P show immunoblots of THP-1 cells treated with 5 μM Compound 1 or 2′-3′cGAMP. [Figure 52F]52A-52P show analysis of IRF3 and NF-kB pathways after treatment with Compound 1. Figures 52A-52P show immunoblots of THP-1 cells treated with 5 μM Compound 1 or 2′-3′cGAMP. [Figure 52G] 52A-52P show analysis of IRF3 and NF-kB pathways after treatment with Compound 1. Figures 52A-52P show immunoblots of THP-1 cells treated with 5 μM Compound 1 or 2′-3′cGAMP. [Figure 52H] 52A-52P show analysis of IRF3 and NF-kB pathways after treatment with Compound 1. Figures 52A-52P show immunoblots of THP-1 cells treated with 5 μM Compound 1 or 2′-3′cGAMP. [Figure 52I] 52A-52P show analysis of IRF3 and NF-kB pathways after treatment with Compound 1. Figures 52A-52P show immunoblots of THP-1 cells treated with 5 μM Compound 1 or 2′-3′cGAMP. [Figure 52J] 52A-52P show analysis of IRF3 and NF-kB pathways after treatment with Compound 1. Figures 52A-52P show immunoblots of THP-1 cells treated with 5 μM Compound 1 or 2′-3′cGAMP. [Figure 52K] 52A-52P show analysis of IRF3 and NF-kB pathways after treatment with Compound 1. Figures 52A-52P show immunoblots of THP-1 cells treated with 5 μM Compound 1 or 2′-3′cGAMP. [Figure 52L] 52A-52P show analysis of IRF3 and NF-kB pathways after treatment with Compound 1. Figures 52A-52P show immunoblots of THP-1 cells treated with 5 μM Compound 1 or 2′-3′cGAMP. [Figure 52M] 52A-52P show analysis of IRF3 and NF-kB pathways after treatment with Compound 1. Figures 52A-52P show immunoblots of THP-1 cells treated with 5 μM Compound 1 or 2′-3′cGAMP. [Figure 52N] 52A-52P show analysis of IRF3 and NF-kB pathways after treatment with Compound 1. Figures 52A-52P show immunoblots of THP-1 cells treated with 5 μM Compound 1 or 2′-3′cGAMP. [Figure 52O]52A-52P show analysis of IRF3 and NF-kB pathways after treatment with Compound 1. Figures 52A-52P show immunoblots of THP-1 cells treated with 5 μM Compound 1 or 2′-3′cGAMP. [Figure 52P] 52A-52P show analysis of IRF3 and NF-kB pathways after treatment with Compound 1. Figures 52A-52P show immunoblots of THP-1 cells treated with 5 μM Compound 1 or 2′-3′cGAMP. [Figure 53A] THP-1 derived macrophages were treated with Compound 1 or DMSO control for 2 hours (Figure 53A), 4 hours (Figure 53B), or 6 hours (Figure 53C) and analyzed for nuclear translocation. Cells were imaged with an IXM (Molecular Devices) at 40x magnification and analyzed using ImageJ. [Figure 53B] THP-1 derived macrophages were treated with Compound 1 or DMSO control for 2 hours (Figure 53A), 4 hours (Figure 53B), or 6 hours (Figure 53C) and analyzed for nuclear translocation. Cells were imaged with an IXM (Molecular Devices) at 40x magnification and analyzed using ImageJ. [Figure 53C] THP-1 derived macrophages were treated with Compound 1 or DMSO control for 2 hours (Figure 53A), 4 hours (Figure 53B), or 6 hours (Figure 53C) and analyzed for nuclear translocation. Cells were imaged with an IXM (Molecular Devices) at 40x magnification and analyzed using ImageJ. [Figure 54A] Figure 54A shows the evaluation of IFN secretion and gene expression after treatment with Compound 1. Figure 54A is a graph showing the fold induction of gene expression in THP-1 cells treated with either 5 μM Compound 1 or 2'3-cGAMP. Gene expression was evaluated by Taqman Assays. Fold induction was calculated using the ΔΔct method. In Figure 54B, THP-1 cells were treated with 1 μM Compound 1, and the secretion of certain cytokines was evaluated by IFN and IL-1 family multiplex ELISA arrays on a Quansys Biosciences (Logan, UT) Q-Plex™ Human Custom. [Figure 54B] Figure 54A shows the evaluation of IFN secretion and gene expression after treatment with Compound 1. Figure 54A is a graph showing the fold induction of gene expression in THP-1 cells treated with either 5 μM Compound 1 or 2'3-cGAMP. Gene expression was evaluated by Taqman Assays. Fold induction was calculated using the ΔΔct method. In Figure 54B, THP-1 cells were treated with 1 μM Compound 1, and the secretion of certain cytokines was evaluated by IFN and IL-1 family multiplex ELISA arrays on a Quansys Biosciences (Logan, UT) Q-Plex™ Human Custom. [Figure 54C] Figure 54D shows the induction of apoptosis-related genes and ISGs by an exemplary compound (Compound 1) compared to 2'3'-cGAMP in A20 mouse B-cell lymphoma tumor cells. In Figure 54D, the higher BAX / BCL2 ratio in cells treated with Compound 1 promotes apoptosis via upregulation of caspase 3. [Figure 54D] Figure 54D shows the induction of apoptosis-related genes and ISGs by an exemplary compound (Compound 1) compared to 2'3'-cGAMP in A20 mouse B-cell lymphoma tumor cells. In Figure 54D, the higher BAX / BCL2 ratio in cells treated with Compound 1 promotes apoptosis via upregulation of caspase 3. [Figure 55A] 1 is a graph showing the induction of various cytokines by Compound 1 in wild-type THP1 cells as determined by multiplex ELISA. [Figure 55B] 1 is a graph showing the induction of various cytokines by Compound 1 in wild-type THP1 cells as determined by multiplex ELISA. [Figure 55C] 1 is a graph showing the induction of various cytokines by Compound 1 in wild-type THP1 cells as determined by multiplex ELISA. [Figure 55D] 1 is a graph showing the induction of various cytokines by Compound 1 in wild-type THP1 cells as determined by multiplex ELISA. [Figure 55E] 1 is a graph showing the induction of various cytokines by Compound 1 in wild-type THP1 cells as determined by multiplex ELISA. [Figure 55F] 1 is a graph showing the induction of various cytokines by Compound 1 in wild-type THP1 cells as determined by multiplex ELISA. [Figure 55G] 1 is a graph showing the induction of various cytokines by Compound 1 in wild-type THP1 cells as determined by multiplex ELISA. [Figure 56A] 1 is a graph showing that an exemplary compound (Compound 1) potently activates IRF-type I and type III IFN responses. [Figure 56B] 1 is a graph showing that an exemplary compound (Compound 1) potently activates IRF-type I and type III IFN responses. [Figure 56C] 1 is a graph showing that an exemplary compound (Compound 1) potently activates IRF-type I and type III IFN responses. [Figure 56D] 1 is a graph showing that an exemplary compound (Compound 1) potently activates IRF-type I and type III IFN responses. [Figure 57] FIG. 1 shows that an exemplary compound (Compound 1) activates human natural killer (NK) cells and induces IFN-γ production. [Figure 58A] In a syngeneic A20 lymphoma model, an exemplary compound (Compound 1) is shown to potently suppress lymphoma tumor growth. [Figure 58B] In a syngeneic A20 lymphoma model, an exemplary compound (Compound 1) is shown to potently suppress lymphoma tumor growth. [Figure 59A] 1 is a graph showing that an exemplary compound (Compound 1) administered in combination with cyclophosphamide results in tumor-free survival in a syngeneic A20 lymphoma mouse model. [Figure 59B]1 is a graph showing that an exemplary compound (Compound 1) administered in combination with cyclophosphamide results in tumor-free survival in a syngeneic A20 lymphoma mouse model. [Figure 59C] 1 is a graph showing that an exemplary compound (Compound 1) administered in combination with cyclophosphamide results in tumor-free survival in a syngeneic A20 lymphoma mouse model. [Figure 59D] 1 is a graph showing that an exemplary compound (Compound 1) administered in combination with cyclophosphamide results in tumor-free survival in a syngeneic A20 lymphoma mouse model. [Figure 60A] In a syngeneic A20 lymphoma mouse model, compound 1 monotherapy and compound 1 combined with cyclophosphamide significantly improved mouse survival. Note that in Figure 60B, VS1 refers to compound 1. [Figure 60B] In a syngeneic A20 lymphoma mouse model, compound 1 monotherapy and compound 1 combined with cyclophosphamide significantly improved mouse survival. Note that in Figure 60B, VS1 refers to compound 1. [Figure 61A] 1 is an image showing immunohistochemistry data from tissues taken from mice treated with Compound 1. The images show that the anti-tumor activity of Compound 1 correlates with the induction of innate and adaptive immune responses. [Figure 61B] 1 is an image showing immunohistochemistry data from tissues taken from mice treated with Compound 1. The images show that the anti-tumor activity of Compound 1 correlates with the induction of innate and adaptive immune responses. [Figure 61C] 1 is an image showing immunohistochemistry data from tissues taken from mice treated with Compound 1. The images show that the anti-tumor activity of Compound 1 correlates with the induction of innate and adaptive immune responses. [Figure 61D] 1 is an image showing immunohistochemistry data from tissues taken from mice treated with Compound 1. The images show that the anti-tumor activity of Compound 1 correlates with the induction of innate and adaptive immune responses. [Figure 62A] In a syngeneic CT26 colon cancer model, an exemplary compound (Compound 1) is shown to be highly effective in suppressing tumor growth. [Figure 62B] In a syngeneic CT26 colon cancer model, an exemplary compound (Compound 1) is shown to be highly effective in suppressing tumor growth. [Figure 63A] In a syngeneic CT26 colon cancer mouse model, compound 1 monotherapy and compound 1 combined with anti-CTLA4 antibody significantly improved mouse survival. Note that in Figure 63B, VS1 refers to compound 1. [Figure 63B] In a syngeneic CT26 colon cancer mouse model, compound 1 monotherapy and compound 1 combined with anti-CTLA4 antibody significantly improved mouse survival. Note that in Figure 63B, VS1 refers to compound 1. [Figure 64] 1 shows that mice found to be tumor-free after treatment with either Compound 1 or Compound 1 + cyclophosphamide do not experience tumor growth compared to controls when the mice are rechallenged with tumor cells (A20 lymphoma tumor challenge test). [Figure 65A] 6A-6H are images showing immunohistochemistry data using an anti-CD38 antibody on tumor tissue collected from mice treated with vehicle (FIGS. 65A-65D) or Compound 1 (FIGS. 65E-65H) in a syngeneic A20 lymphoma model. The images show that Compound 1 induces CD8 T cell migration to the tumor site. [Figure 65B] 6A-6H are images showing immunohistochemistry data using an anti-CD38 antibody on tumor tissue collected from mice treated with vehicle (FIGS. 65A-65D) or Compound 1 (FIGS. 65E-65H) in a syngeneic A20 lymphoma model. The images show that Compound 1 induces CD8 T cell migration to the tumor site. [Figure 65C]6A-6H are images showing immunohistochemistry data using an anti-CD38 antibody on tumor tissue collected from mice treated with vehicle (FIGS. 65A-65D) or Compound 1 (FIGS. 65E-65H) in a syngeneic A20 lymphoma model. The images show that Compound 1 induces CD8 T cell migration to the tumor site. [Figure 65D] 6A-6H are images showing immunohistochemistry data using an anti-CD38 antibody on tumor tissue collected from mice treated with vehicle (FIGS. 65A-65D) or Compound 1 (FIGS. 65E-65H) in a syngeneic A20 lymphoma model. The images show that Compound 1 induces CD8 T cell migration to the tumor site. [Figure 65E] 6A-6H are images showing immunohistochemistry data using an anti-CD38 antibody on tumor tissue collected from mice treated with vehicle (FIGS. 65A-65D) or Compound 1 (FIGS. 65E-65H) in a syngeneic A20 lymphoma model. The images show that Compound 1 induces CD8 T cell migration to the tumor site. [Figure 65F] 6A-6H are images showing immunohistochemistry data using an anti-CD38 antibody on tumor tissue collected from mice treated with vehicle (FIGS. 65A-65D) or Compound 1 (FIGS. 65E-65H) in a syngeneic A20 lymphoma model. The images show that Compound 1 induces CD8 T cell migration to the tumor site. [Figure 65G] 6A-6H are images showing immunohistochemistry data using an anti-CD38 antibody on tumor tissue collected from mice treated with vehicle (FIGS. 65A-65D) or Compound 1 (FIGS. 65E-65H) in a syngeneic A20 lymphoma model. The images show that Compound 1 induces CD8 T cell migration to the tumor site. [Figure 65H] 6A-6H are images showing immunohistochemistry data using an anti-CD38 antibody on tumor tissue collected from mice treated with vehicle (FIGS. 65A-65D) or Compound 1 (FIGS. 65E-65H) in a syngeneic A20 lymphoma model. The images show that Compound 1 induces CD8 T cell migration to the tumor site. [Figure 66A]66A-66H are images showing immunohistochemistry data using an anti-granzyme B antibody on tumor tissues from mice treated with vehicle (FIGS. 66A-66D) or Compound 1 (FIGS. 66E-66H) in a syngeneic A20 lymphoma model. The images show that Compound 1 induces NK cell migration to the tumor site. [Figure 66B] 66A-66H are images showing immunohistochemistry data using an anti-granzyme B antibody on tumor tissues from mice treated with vehicle (FIGS. 66A-66D) or Compound 1 (FIGS. 66E-66H) in a syngeneic A20 lymphoma model. The images show that Compound 1 induces NK cell migration to the tumor site. [Figure 66C] 66A-66H are images showing immunohistochemistry data using an anti-granzyme B antibody on tumor tissues from mice treated with vehicle (FIGS. 66A-66D) or Compound 1 (FIGS. 66E-66H) in a syngeneic A20 lymphoma model. The images show that Compound 1 induces NK cell migration to the tumor site. [Figure 66D] 66A-66H are images showing immunohistochemistry data using an anti-granzyme B antibody on tumor tissues from mice treated with vehicle (FIGS. 66A-66D) or Compound 1 (FIGS. 66E-66H) in a syngeneic A20 lymphoma model. The images show that Compound 1 induces NK cell migration to the tumor site. [Figure 66E] 66A-66H are images showing immunohistochemistry data using an anti-granzyme B antibody on tumor tissues from mice treated with vehicle (FIGS. 66A-66D) or Compound 1 (FIGS. 66E-66H) in a syngeneic A20 lymphoma model. The images show that Compound 1 induces NK cell migration to the tumor site. [Figure 66F] 66A-66H are images showing immunohistochemistry data using an anti-granzyme B antibody on tumor tissues from mice treated with vehicle (FIGS. 66A-66D) or Compound 1 (FIGS. 66E-66H) in a syngeneic A20 lymphoma model. The images show that Compound 1 induces NK cell migration to the tumor site. [Figure 66G]66A-66H are images showing immunohistochemistry data using an anti-granzyme B antibody on tumor tissues from mice treated with vehicle (FIGS. 66A-66D) or Compound 1 (FIGS. 66E-66H) in a syngeneic A20 lymphoma model. The images show that Compound 1 induces NK cell migration to the tumor site. [Figure 66H] 66A-66H are images showing immunohistochemistry data using an anti-granzyme B antibody on tumor tissues from mice treated with vehicle (FIGS. 66A-66D) or Compound 1 (FIGS. 66E-66H) in a syngeneic A20 lymphoma model. The images show that Compound 1 induces NK cell migration to the tumor site. [Figure 67A] 67A-67H are images showing immunohistochemistry data using anti-F4 / 80 antibody on tumor tissue collected from mice treated with vehicle (FIGS. 67A-67D) or Compound 1 (FIGS. 67E-67H) in a syngeneic A20 lymphoma model. The images show that Compound 1 induces macrophage migration to the tumor site. [Figure 67B] 67A-67H are images showing immunohistochemistry data using anti-F4 / 80 antibody on tumor tissue collected from mice treated with vehicle (FIGS. 67A-67D) or Compound 1 (FIGS. 67E-67H) in a syngeneic A20 lymphoma model. The images show that Compound 1 induces macrophage migration to the tumor site. [Figure 67C] 67A-67H are images showing immunohistochemistry data using anti-F4 / 80 antibody on tumor tissue collected from mice treated with vehicle (FIGS. 67A-67D) or Compound 1 (FIGS. 67E-67H) in a syngeneic A20 lymphoma model. The images show that Compound 1 induces macrophage migration to the tumor site. [Figure 67D] 67A-67H are images showing immunohistochemistry data using anti-F4 / 80 antibody on tumor tissue collected from mice treated with vehicle (FIGS. 67A-67D) or Compound 1 (FIGS. 67E-67H) in a syngeneic A20 lymphoma model. The images show that Compound 1 induces macrophage migration to the tumor site. [Figure 67E]67A-67H are images showing immunohistochemistry data using anti-F4 / 80 antibody on tumor tissue collected from mice treated with vehicle (FIGS. 67A-67D) or Compound 1 (FIGS. 67E-67H) in a syngeneic A20 lymphoma model. The images show that Compound 1 induces macrophage migration to the tumor site. [Figure 67F] 67A-67H are images showing immunohistochemistry data using anti-F4 / 80 antibody on tumor tissue collected from mice treated with vehicle (FIGS. 67A-67D) or Compound 1 (FIGS. 67E-67H) in a syngeneic A20 lymphoma model. The images show that Compound 1 induces macrophage migration to the tumor site. [Figure 67G] 67A-67H are images showing immunohistochemistry data using anti-F4 / 80 antibody on tumor tissue collected from mice treated with vehicle (FIGS. 67A-67D) or Compound 1 (FIGS. 67E-67H) in a syngeneic A20 lymphoma model. The images show that Compound 1 induces macrophage migration to the tumor site. [Figure 67H] 67A-67H are images showing immunohistochemistry data using anti-F4 / 80 antibody on tumor tissue collected from mice treated with vehicle (FIGS. 67A-67D) or Compound 1 (FIGS. 67E-67H) in a syngeneic A20 lymphoma model. The images show that Compound 1 induces macrophage migration to the tumor site. [Figure 68A] 1 shows administration of Compound 1 to a panel of normal cell lines, demonstrating that Compound 1 is non-cytotoxic. [Figure 68B] 1 shows administration of Compound 1 to a panel of normal cell lines, demonstrating that Compound 1 is non-cytotoxic. [Figure 68C] 1 shows administration of Compound 1 to a panel of normal cell lines, demonstrating that Compound 1 is non-cytotoxic. [Figure 68D] 1 shows administration of Compound 1 to a panel of normal cell lines, demonstrating that Compound 1 is non-cytotoxic. [Figure 68E] 1 shows administration of Compound 1 to a panel of normal cell lines, demonstrating that Compound 1 is non-cytotoxic. [Figure 68F]1 shows administration of Compound 1 to a panel of normal cell lines, demonstrating that Compound 1 is non-cytotoxic. [Figure 68G] 1 shows administration of Compound 1 to a panel of normal cell lines, demonstrating that Compound 1 is non-cytotoxic. [Figure 69A] Figure 69 shows that palmitoylation of STING is involved in Compound 1-induced activation of NF-κB (Figures 69A-69B) and IRF-type I interferon responses in THP1 cells (Figures 69C-69D). [Figure 69B] Figure 69 shows that palmitoylation of STING is involved in Compound 1-induced activation of NF-κB (Figures 69A-69B) and IRF-type I interferon responses in THP1 cells (Figures 69C-69D). [Figure 69C] Figure 69 shows that palmitoylation of STING is involved in Compound 1-induced activation of NF-κB (Figures 69A-69B) and IRF-type I interferon responses in THP1 cells (Figures 69C-69D). [Figure 69D] Figure 69 shows that palmitoylation of STING is involved in Compound 1-induced activation of NF-κB (Figures 69A-69B) and IRF-type I interferon responses in THP1 cells (Figures 69C-69D). [Figure 70] 1 is a graph showing that intraperitoneal administration of Compound 1 significantly reduces tumor burden in a syngeneic mouse metastatic breast cancer model described in Example 12. [Figure 71] 1 is a graph showing the results of an oral dosing study, showing that all participating subjects were within the acceptable weight range. [Figure 72] 1 is a graph showing the antitumor activity of Compound 1, Compound X, and Compound 21 in a syngeneic mouse A20 lymphoma model. All compounds demonstrated significant tumor growth inhibition compared to vehicle. [Figure 73A]Figure 73A shows the ventral antitumor activity of Compound 1 when administered intratumorally in a CT26 colon cancer model. Figure 73A shows the tumor burden of the left ventral tumor over 13 days after initial treatment, and Figure 73B shows the tumor burden of the right ventral tumor. Compound 1 showed significant tumor growth inhibition compared to vehicle. [Figure 73B] Figure 73A shows the ventral antitumor activity of Compound 1 when administered intratumorally in a CT26 colon cancer model. Figure 73A shows the tumor burden of the left ventral tumor over 13 days after initial treatment, and Figure 73B shows the tumor burden of the right ventral tumor. Compound 1 showed significant tumor growth inhibition compared to vehicle. [Figure 74] 1 is a graph showing the effect on tumor growth in a CT26 colon cancer model of a dose of vehicle and 10 μg, 30 μg, and 100 μg of Compound 1. Compound 1 showed significant tumor growth inhibition at all three doses compared to vehicle. [Figure 75] 1 is a graph showing the effect of Compound 1 and vehicle on tumor growth in a 4T1 breast cancer model. Compound 1 showed significant tumor growth inhibition compared to vehicle. [Figure 76A] Figure 1 shows the percentage induction of CD8+ T cells, CD4+ T cells, and MDSCs by Compound 1 in the spleen, lymph nodes, and blood after 19 days as measured by flow cytometry. Compound 1 showed an increase in CD8+ T cells, CD4+ T cells, and MDSCs compared to vehicle. [Figure 76B] Figure 1 shows the percentage induction of CD8+ T cells, CD4+ T cells, and MDSCs by Compound 1 in the spleen, lymph nodes, and blood after 19 days as measured by flow cytometry. Compound 1 showed an increase in CD8+ T cells, CD4+ T cells, and MDSCs compared to vehicle. [Figure 76C] Figure 1 shows the percentage induction of CD8+ T cells, CD4+ T cells, and MDSCs by Compound 1 in the spleen, lymph nodes, and blood after 19 days as measured by flow cytometry. Compound 1 showed an increase in CD8+ T cells, CD4+ T cells, and MDSCs compared to vehicle. [Figure 76D] Figure 1 shows the percentage induction of CD8+ T cells, CD4+ T cells, and MDSCs by Compound 1 in the spleen, lymph nodes, and blood after 19 days as measured by flow cytometry. Compound 1 showed an increase in CD8+ T cells, CD4+ T cells, and MDSCs compared to vehicle. [Figure 77] 1 is a graph showing the antitumor activity of vehicle Compound 1, Compound 1A (an isomer of Compound 1), and Compound 21. The graph shows that Compound 1, Compound 1A, and Compound 21 inhibit mouse A20 B-cell lymphoma tumor cells. [Figure 78] 1 is a Kaplan-Meier plot showing that Compound 1, Compound 1A, and Compound 21 significantly improve mouse survival in an A26 lymphoma model. [Figure 79] 1 is a graph showing the effect of intratumoral administration of Compound 1, Compound 21, and Compound 25 on tumor growth in a CT26 colon cancer model. Compound 1, Compound 21, and Compound 25 showed significant tumor growth inhibition compared to vehicle. [Figure 80] Graph showing the effect of intratumoral administration of vehicle, vehicle, and ethanol on tumor growth. Compound 1, Compound 1A, and Compound 1A. Compound 1, Compound 21, and Compound 25 in a CT26 colon cancer model showed significant tumor growth inhibition compared to vehicle. [Figure 81A] 81A-81B are graphs showing the stability of Compound 1 in rabbit serum (FIG. 81A) and human microsomes (FIG. 81B). [Figure 81B] 81A-81B are graphs showing the stability of Compound 1 in rabbit serum (FIG. 81A) and human microsomes (FIG. 81B). [Figure 82] 10A-10C are luminescence images showing the effect of intraperitoneal administration of Compound 1 on tumor growth in a 4T1 breast cancer syngeneic mouse model. Compound 1 demonstrated significant tumor growth inhibition at all three doses compared to vehicle. DETAILED DESCRIPTION OF THE INVENTION

[0044] The present invention relates to a method for activating and / or inducing the expression of a PRR (e.g., STING) in a subject, particularly for the treatment of a proliferative disease (e.g., cancer). In some embodiments, the method comprises administering a compound of formula (I) or a pharmaceutically acceptable salt thereof. It should be noted that the induction of any PRR by these compounds can stimulate interferon and / or NF-κB production, which can induce the expression of various PRRs that are inducible genes through a feedback mechanism.

[0045] definition As used herein, the articles "a" and "an" refer to one or to more than one (ie, to at least one) of the grammatical object of the article.

[0046] "About" and "approximately" generally refer to an acceptable degree of error for the quantity measured, given the nature or precision of the measurement. Exemplary degrees of error are within 20 percent (%), typically within 10%, and more typically within 5% of a given value or range of values.

[0047] As used herein, the terms "obtain" or "obtaining," as these terms are used herein, refer to possession of a physical entity (e.g., a sample, e.g., a blood sample or liver biopsy sample) or obtaining a value, e.g., a numerical value, by "directly obtaining" or "indirectly obtaining" a physical entity or value. "Directly obtaining" means performing a process (e.g., an analytical method) to obtain the physical entity or value. "Indirectly obtaining" refers to receiving a physical entity or value from another party or source (e.g., a non-party laboratory that directly obtained the physical entity or value). Directly obtaining a value includes performing a process that involves a physical change of a sample or another substance, e.g., performing an analytical process that involves a physical change of a substance, e.g., a sample, performing an analytical method, e.g., a method described herein, e.g., by analyzing a sample of a bodily fluid such as blood by mass spectrometry, e.g., LC-MS.

[0048] As used herein, the term "induce" or "induction of" refers to an increase or enhancement of function, for example, an increase or enhancement of expression of a pattern recognition receptor (e.g., STING). In some embodiments, "induction of PRR expression" refers to an induction of transcription of a PRR RNA, e.g., STING RNA (e.g., mRNA, e.g., an increase or enhancement thereof), or a PRR protein, e.g., STING protein (e.g., an increase or enhancement thereof). In some embodiments, induction of PRR expression (e.g., STING expression) refers, for example, to an increase or enhancement of the concentration of a PRR RNA, e.g., STING RNA (e.g., mRNA), or STING protein, in a cell. In some embodiments, induction of PRR expression (e.g., STING expression) refers, for example, to an increase in the copy number of a PRR RNA, e.g., STING RNA (e.g., mRNA), or a PRR protein, e.g., STING protein, in a cell. In some embodiments, inducing expression of a PRR (e.g., STING) can refer to the initiation of PRR RNA (e.g., STING RNA (e.g., mRNA)) or transcription, or the initiation of PRR protein (e.g., STING protein) translation. In some embodiments, inducing expression of a PRR (e.g., STING) can refer to an increase in the rate of PRR RNA (e.g., STING RNA (e.g., mRNA)) transcription, or an increase in the rate of PRR protein (e.g., STING protein) expression.

[0049] As used herein, the term "activate" or "activation" refers to, for example, the stimulation or induction of a downstream pathway, e.g., the function of a downstream signaling pathway. In some embodiments, activation of a pattern recognition receptor (PRR) (e.g., STING) refers to the stimulation of a specific protein or pathway, for example, through interaction with a downstream signaling partner (e.g., IFN-β promoter stimulator 1 (IPS-1), IRF3, IRF7, NF-κB, interferons (e.g., IFN-α or IFN-β), and / or cytokines). In some embodiments, activation is distinct from the induction of expression of a PRR. In some embodiments, a PRR may be activated without resulting in the induction of PRR expression (e.g., expression of STING). In some embodiments, activation may include the induction of expression of a PRR (e.g., STING). In some embodiments, activation of a PRR may induce induction of expression of a PRR (e.g., STING) by about 0.1%, about 0.5%, about 1%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, or more compared to a reference standard (e.g., basal expression level of a PRR (e.g., STING)).

[0050] As used herein, an amount of a compound, complex, or substance effective to treat a disorder (e.g., a disorder described herein), a "therapeutically effective amount," "effective amount," or "effective course" refers to the amount of a compound, substance, or composition that, upon single or multiple administrations to a subject, is effective in treating the subject or in curing, alleviating, ameliorating, or improving a subject suffering from a disorder beyond that expected in the absence of such treatment.

[0051] The terms "prevent" or "preventing," as used in the context of a disorder or disease, refer to the administration of an agent to a subject such that the onset of at least one symptom of the disorder or disease is delayed compared to the onset that would be observed in the absence of administration of such agent, e.g., administration of a compound of the present disclosure (e.g., a compound of Formula (I)) to a subject.

[0052] As used herein, the term "reference treatment" or "reference standard" refers to a standardized level or standardized treatment used as a basis for comparison. In some embodiments, the reference standard or reference treatment is an art-recognized, well-known, or well-characterized standard or treatment. In some embodiments, the reference standard describes the results of a method described herein. In some embodiments, the reference standard describes, e.g., the level of a marker (e.g., the level of induction of a PRR, e.g., STING) in a subject or sample, e.g., prior to the initiation of treatment, e.g., with a compound or composition described herein. In some embodiments, the reference standard describes a measure of the presence, progression, or severity of a disease or a symptom thereof, e.g., prior to the initiation of treatment, e.g., with a compound or composition described herein.

[0053] As used herein, the term "subject" is intended to include humans and non-human animals. Exemplary human subjects include human patients suffering from a disorder, e.g., a disorder described herein, or a normal subject. The term "non-human animal" includes all vertebrates, e.g., non-mammals (e.g., chickens, amphibians, reptiles), as well as mammals such as, for example, non-human primates, domesticated animals, and / or agriculturally useful animals, e.g., sheep, dogs, cats, pigs, etc. In an exemplary embodiment of the present disclosure, the subject is a woodchuck (e.g., eastern woodchuck (Marmota monax)).

[0054] As used herein, the term "treat" or "treating" a subject suffering from a disorder or disease refers to subjecting the subject to a treatment regimen, e.g., administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a composition comprising a compound of Formula (I) or a pharmaceutically acceptable salt thereof, so that at least one symptom of the disorder or disease is cured, healed, alleviated, relieved, altered, remedied, ameliorated, or improved. Treating includes administering an amount effective to alleviate, relieve, alter, remedy, improve, or affect the disorder or disease, or the symptoms of the disorder or disease. Treatment may inhibit the deterioration or worsening of the symptoms of the disorder or disease.

[0055] As used herein, the term "Cmd" refers to "compound" or "Compound," all of which terms are used interchangeably.

[0056] Numerous ranges, e.g., ranges for amounts of drug administered per day, are provided herein. In some embodiments, the ranges include both endpoints. In other embodiments, the ranges exclude one or both endpoints. For example, a range can exclude a lower limit. Thus, in such an embodiment, a range of 250 to 400 mg / day, excluding the lower limit, would encompass amounts greater than 250 that are up to 400 mg / day.

[0057] definition As used herein, the term "alkyl" refers to any alkyl group selected from C1 to C6. 12 Alkyl, C1-C 10Alkyl refers to a monovalent saturated straight or branched chain hydrocarbon, such as a straight or branched group of 1 to 12, 1 to 10, or 1 to 6 carbon atoms, referred to as C1-C6 alkyl. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, sec-pentyl, isopentyl, tert-butyl, n-pentyl, neopentyl, n-hexyl, sec-hexyl, and the like.

[0058] The terms "alkenyl" and "alkynyl" are art-recognized and refer to unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but which contain at least one double or triple bond respectively. Exemplary alkenyl groups include, but are not limited to, -CH=CH2 and -CH2CH=CH2.

[0059] The term "alkylene" refers to a diradical of an alkyl group.

[0060] The terms "alkenylene" and "alkynylene" refer to diradicals of alkenyl and alkynyl groups, respectively.

[0061] The term "methylene unit" refers to a divalent -CH2- group present in an alkyl, alkenyl, alkynyl, alkylene, alkenylene, or alkynylene moiety.

[0062] The term "carbocyclic ring system," as used herein, means a monocyclic, or a fused, spiro-fused, and / or bridged bicyclic or polycyclic hydrocarbon ring system, in which each ring is either fully saturated or contains one or more units of unsaturation, but none of the rings is aromatic.

[0063] The term "carbocyclyl" refers to a group of carbocyclic ring systems. Representative carbocyclyl groups include cycloalkyl groups (e.g., cyclopentyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.) and cycloalkenyl groups (e.g., cyclopentenyl, cyclohexenyl, cyclopentadienyl, etc.).

[0064] The term "aromatic ring system" is art-recognized and refers to a monocyclic, bicyclic, or polycyclic hydrocarbon ring system in which at least one ring is aromatic.

[0065] The term "aryl" refers to a radical of an aromatic ring system. Representative aryl groups include fully aromatic ring systems such as phenyl, naphthyl, and anthracenyl, as well as ring systems in which an aromatic carbocyclic ring is fused to one or more non-aromatic carbocyclic rings, such as indanyl, phthalimidyl, naphthymidyl, or tetrahydronaphthyl.

[0066] The term "heteroalkyl" refers to an "alkyl" moiety in which at least one carbon atom has been replaced with a heteroatom such as O, S, or N.

[0067] The term "heteroaromatic ring system" is art-recognized and refers to a monocyclic, bicyclic, or polycyclic ring system in which at least one ring is aromatic and contains a heteroatom, and none of the other rings are heterocyclyl (defined below). In certain cases, an aromatic, heteroatom-containing ring contains 1, 2, 3, or 4 independently selected ring heteroatoms in such ring.

[0068] The term "heteroaryl" refers to a group of heteroaromatic ring systems. Representative heteroaryl groups include (i) ring systems in which the rings each contain a heteroatom and are aromatic, e.g., imidazolyl, oxazolyl, thiazolyl, triazolyl, pyrrolyl, furanyl, thiophenylpyrazolyl, pyridinyl, pyrazinyl, pyridazinyl, pyrimidinyl, indolizinyl, purinyl, naphthyridinyl, and pteridinyl; (ii) rings in which the rings each are aromatic or carbocyclyl, in which at least one aromatic ring contains a heteroatom and at least one other ring is a hydrocarbon ring or a heterocyclic ring, e.g., indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indyl, and the like. The ring system includes (iii) zolyl, benzimidazolyl, benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, pyrido[2,3-b]-1,4-oxazin-3(4H)-one, 5,6,7,8-tetrahydroquinolinyl and 5,6,7,8-tetrahydroisoquinolinyl, and (iii) ring systems in which each ring is aromatic or carbocyclyl, and at least one aromatic ring shares a bridgehead heteroatom with another aromatic ring, such as 4H-quinolizinyl. In certain embodiments, heteroaryl is a monocyclic or bicyclic ring, and each of these rings contains 5 or 6 ring atoms, of which 1, 2, 3 or 4 of these ring atoms are heteroatoms independently selected from N, O and S.

[0069] The term "heterocyclic ring system" refers to monocyclic, or fused, spiro-fused, and / or bridged bicyclic and polycyclic ring systems in which at least one ring is saturated or partially unsaturated (but not aromatic) and contains a heteroatom. The heterocyclic ring system can be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure, and any of the ring atoms can be optionally substituted.

[0070] The term "heterocyclyl" refers to a group of heterocyclic ring systems. Representative heterocyclyls include (i) ring systems in which all rings are non-aromatic and at least one ring contains a heteroatom, such as tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, pyrrolidonyl, piperidinyl, pyrrolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl; (ii) ring systems in which at least one ring is non-aromatic and at least one ring contains a heteroatom, such as tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, pyrrolidinyl, piperidinyl, pyrrolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl; and (iii) ring systems in which at least one ring is non-aromatic and contains a heteroatom and at least one other ring is aromatic and contains a heteroatom, such as 3,4-dihydro-1H-pyrano[4,3-c]pyridine and 1,2,3,4-tetrahydro-2,6-naphthyridine. In certain embodiments, the heterocyclyl is a monocyclic or bicyclic ring, wherein each of the rings contains 3 to 7 ring atoms, and 1, 2, 3, or 4 of the ring atoms are heteroatoms independently selected from N, O, and S.

[0071] The term "saturated heterocyclyl" refers to a heterocyclic group in which all rings are saturated, for example, tetrahydrofuran, tetrahydro-2H-pyran, pyrrolidine, piperidine, and piperazine.

[0072] "Partially unsaturated" refers to a group that contains at least one double or triple bond. "Partially unsaturated" ring systems are also intended to encompass rings with multiple sites of unsaturation, but are not intended to include aromatic groups (e.g., aryl or heteroaryl groups) as defined herein. Similarly, "saturated" refers to a group that contains no double or triple bonds, i.e., all single bonds.

[0073] As used herein, the term "nucleobase" refers to a nucleoside, a nitrogen-containing biological compound found attached to the sugar in the basic building blocks of deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). The primary, or naturally occurring, nucleobases are cytosine (DNA and RNA), guanine (DNA and RNA), adenine (DNA and RNA), thymine (DNA), and uracil (RNA), abbreviated as C, G, A, T, and U, respectively. Because A, G, C, and T occur in DNA, these molecules are called DNA bases, while A, G, C, and U are called RNA bases. Adenine and guanine belong to the double-ring class of molecules called purines (abbreviated R). Cytosine, thymine, and uracil are all pyrimidines. Other nucleobases that do not function as a normal part of the genetic code are called non-natural.

[0074] As described herein, compounds of the present disclosure may contain "optionally substituted" moieties. Generally, the term "substituted," whether preceded by the term "optionally," means that one or more hydrogens of the specified moiety have been replaced with a suitable substituent. Unless otherwise specified, an "optionally substituted" group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituents may be the same or different at each position. Combinations of substituents contemplated under the present disclosure are preferably those that result in the formation of stable or chemically feasible compounds. The term "stable," as used herein, refers to a compound that remains substantially unchanged when subjected to conditions that allow for the production, detection, and, in certain embodiments, recovery, purification, and use of the compound for one or more of the purposes disclosed herein.

[0075] As used herein, the definition of each expression, e.g., alkyl, m, n, etc., when this expression occurs more than once in any structure, is intended to be independent of the definition of this expression elsewhere in the same structure.

[0076] As described herein, compounds of the present disclosure may contain "optionally substituted" moieties. Generally, the term "substituted," whether preceded by the term "optionally," means that one or more hydrogens of the specified moiety have been replaced with a suitable substituent. Unless otherwise specified, an "optionally substituted" group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituents may be the same or different at each position. Combinations of substituents contemplated under the present disclosure are preferably those that result in the formation of stable or chemically feasible compounds. The term "stable," as used herein, refers to a compound that remains substantially unchanged when subjected to conditions that allow for the production, detection, and, in certain embodiments, recovery, purification, and use of the compound for one or more of the purposes disclosed herein.

[0077] Pattern Recognition Receptors The present disclosure provides a method for activating and inducing PRR expression (e.g., STING expression) in a subject, e.g., a subject suffering from a proliferative disease (e.g., cancer). Pattern recognition receptors (PRRs) are a broad class of proteins that recognize pathogen-associated molecular patterns (PAMPs) conserved within pathogenic invaders. PAMPs are typically products of biosynthetic pathways essential for pathogen survival and / or infectivity, such as lipopolysaccharides, glycoproteins, and nucleic acids. Recognition of PAMPs by their cognate PRRs activates signaling pathways that result in the production of immune defense factors, such as pro- and anti-inflammatory cytokines, type I interferons (IFN-α, IFN-β), and / or interferon-stimulated genes (ISGs). It is also well known that the induction of innate immune signaling results in the activation of T cell responses and the induction of adaptive immunity. These downstream immune effects are essential for the elimination of viruses via apoptosis and killing of infected cells through cytotoxic T lymphocytes, as well as other defense mechanisms. It is also well known that interferons act on ISREs (interferon response elements), which can induce the production of ISGs, which play an important role in antiviral cellular defense.

[0078] Stimulator of interferon genes (STING) is a cytosolic microbial DNA sensor that has been shown to be particularly sensitive to double-stranded DNA and cyclic dinucleotides (e.g., cyclic di-GMP) (Burdette, DL and Vance, RE (2013) Nat Immunol 14:19-26). Two STING molecules form homodimers mediated by an α-helix present in the C-terminal dimerization domain, and molecular binding studies have revealed that each STING dimer binds one molecule of microbial nucleic acid, e.g., DNA or cyclic dinucleotides. Upon ligand binding, STING activates the innate immune response through interactions with RIG-I and IPS-1, resulting in interferon production (e.g., IFN-α and IFN-β) and other downstream signaling events. Since its discovery, STING has been shown to function as an important sensor of viruses (e.g., adenovirus, herpes simplex virus, hepatitis B virus, vesicular stomatitis virus, and hepatitis C virus), bacteria (e.g., Listeria monocytogenes, Legionella pneumophila, and Mycobacterium tuberculosis), and protozoa (Plasmodium falciparum and Plasmodium berghei). Furthermore, STING has been shown to play a key role in the innate immune response to tumor antigens, driving dendritic cell activation and subsequent T cell priming in several cancers (Woo, SR et al. Trends in Immunol (2015) 36:250-256).

[0079] Another class of PRRs includes RIG-I, the first member of a family of PRRs called RIG-I-like receptors (RLRs), which primarily detect RNA from exogenous sources. RIG-I is an important sensor of microbial infection (e.g., viral infection) in most cells and is constitutively expressed at low levels in the cytosol. After ligand binding, RIG-I expression is rapidly enhanced, resulting in increased RIG-I concentration in the cell (Jensen, S. and Thomsen, ARJ Virol (2012) 86:2900-2910; Yoneyama M. et al. Nat Immunol (2004) 5:730-737). RIG-I is an ATP-dependent helicase containing a central DExD / H-box ATPase domain and a tandem N-terminal caspase recruitment domain (CARD) that mediates downstream signaling. The C-terminus of RIG-I contains a ssRNA / dsRNA-binding domain that acts to silence CARD function at the N-terminus when unbound. Without wishing to be bound by theory, upon recognition of the target RNA structure, two N-terminal CARDs are exposed, allowing interaction with downstream binding partners CARDs and with mitochondrial antiviral signaling molecule (MAVS) and IFN-β promoter stimulator factor 1 (IPS-1), also known as CARDIF. This interaction then triggers further downstream signaling, such as induction of IRF3, IRF7, NF-κB, IFN, and cytokine production, which culminates in the initiation of a host immune response.

[0080] Other RLRs, including MDA5, LGP2, and RNase L, are homologous to RIG-I and function in a similar manner. MDA5 is highly homologous to RIG-I and has been shown to be crucial for inducing cytokine responses during infection with picornaviruses (e.g., encephalomyocarditis virus (EMCV), Taylor virus, and mengovirus), Sendai virus, rabies virus, West Nile virus, rabies virus, rotavirus, mouse hepatitis virus, and murine norovirus. LPG2 lacks the CARD domain found in RIG-I and MDA5, which directly interacts with IPS-1 to initiate downstream signaling. As such, LPG2 is thought to act as a regulator of innate immune responses together with other CARD-bearing RLRs, such as RIG-I and MDA5.

[0081] Another class of PRRs encompasses the nucleotide-binding receptor and oligomerization domain (NOD)-like receptor, or NLR, family (Caruso, R. et al., Immunity (2014) 41:898-908), which includes the microbial sensor NOD2. NOD2 is composed of an N-terminal CARD, a centrally located nucleotide-binding oligomerization domain, and a C-terminal leucine-rich repeat domain responsible for binding microbial PAMPs such as bacterial peptidoglycan fragments and microbial nucleic acids. Ligand binding is thought to activate NOD2, driving its interaction with the CARD-containing kinase RIPK2, which in turn activates numerous downstream proteins, including NF-κB, MAPK, IRF7, and IRF3, the latter of which results in the induction of type 1 interferon. NOD2 is expressed in a diverse set of cell types, including macrophages, dendritic cells, Paneth cells, epithelial cells (e.g., lung epithelial cells, intestinal epithelium), and osteoblasts. NOD2 binds to protozoa (e.g., Toxoplasma gondii and rodent malaria parasite Plasmodium berghei), bacteria (e.g., Bacillus anthracis, Borrelia burgdorferi, Burkholderia pseudomallei, Helicobacter hepaticus, Legionella pneumophilia, Mycobacterium tuberculosis, Propionibacterium acnes, Porphyromonas gingivalis, Salmonella enterica, and Streptococcus pneumoniae). They have been established as sensors of infection by various pathogenic invaders, including bacteria (e.g., respiratory syncytial virus and murine norovirus-1), and viruses (e.g., respiratory syncytial virus and murine norovirus-1) (Moreira, LO and Zamboni, DS Front Immunol (2012) 3:1-12).Recent studies have shown that mutations in NOD2 can contribute to inflammatory diseases such as Crohn's disease, resulting in an abnormal inflammatory response upon stimulation.

[0082] compound The present disclosure features compounds and methods for inducing PRR expression (e.g., STING expression) in a subject (e.g., a subject suffering from a proliferative disease, e.g., cancer), comprising administering a compound of formula (I) or a prodrug thereof, or a pharmaceutically acceptable salt thereof.

[0083] In some embodiments, the disclosure features compounds of Formula (I) in which the 3'-OH terminus of one nucleoside is linked to the 5'-OH of a second nucleoside through a bond as shown. In some other embodiments, the 2'-OH terminus of one nucleoside may be linked to the 5'-OH of a second nucleoside through a bond.

[0084] In some embodiments, the compound has formula (I): [ka] (wherein Z is either S or O, and B 1 and B 2 each of X is independently a purinyl nucleobase or a pyrimidinyl nucleobase; 1 and X 2 each independently is O or S, and Y 1 and Y 2 each independently represents O, S, or NR 5 and L 1 and L 2 each is independently absent, C1-C6 alkyl, or C1-C6 heteroalkyl, where alkyl and heteroalkyl are each optionally selected from R 6 is substituted with R 1 and R 2 each independently represents hydrogen, halo, —CN, C1-C 20 alkyl (e.g., C1-C6 alkyl), or OR 7 and R3 and R 4 each independently represents hydrogen, C1 to C 20 Alkyl (e.g., C1-C6 alkyl), C1-C 20 Heteroalkyl (e.g., C1-C6 heteroalkyl), OC(O)OC1-C 20 alkyl (e.g., C1-C6 alkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each optionally contain one or more R 8 is substituted with R 5 is hydrogen or C1-C 20 alkyl (e.g., C1-C6 alkyl), and R 6 Halo, -CN, C1~C 20 alkyl (e.g., C1-C6 alkyl), OR 7 , oxo, cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl are each optionally joined by one or more R 9 is substituted with R 7 is hydrogen, C1 to C 20 alkyl (e.g., C1-C6 alkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each optionally contain one or more R 9 is substituted with R 8 are each independently C1 to C 20 Alkyl (e.g., C1-C6 alkyl), C1-C 20 Heteroalkyl, C(O)-C1-C 20 Alkyl, OC(O)-C1-C 20 Alkyl (e.g., C1-C6 alkyl), C(O)O-C1-C 20 Alkyl (e.g., C1-C6 alkyl), OC(O)O-C1-C 20 Alkyl (e.g., C1-C6 alkyl), C(O)N(R 5 )-C1~C 20 Alkyl (e.g., C1-C6 alkyl), N(R5 )C(O)-C1~C 20 Alkyl (e.g., C1-C6 alkyl), OC(O)N(R 5 )-C1~C 20 Alkyl (e.g., C1-C6 alkyl), O-aryl, O-heteroaryl, C(O)-aryl, C(O)-heteroaryl, OC(O)-aryl, C(O)O-aryl, OC(O)-heteroaryl, C(O)O-heteroaryl, C(O)O-aryl, C(O) O-heteroaryl, C(O)N(R 5 )-aryl, C(O)N(R 5 )-heteroaryl, N(R 5 )C(O)-aryl, N(R 5 )2C(O)-aryl, or N(R 5 )C(O)-heteroaryl, S(O)N(R 5 )-aryl, wherein alkyl, heteroalkyl, aryl, and heteroaryl are each optionally joined by one or more R 9 is replaced by R 9 are each independently C1 to C 20 Alkyl, O-C1-C 20 Alkyl, C1-C 20 heteroalkyl, halo, —CN, OH, oxo, aryl, heteroaryl, O-aryl, or O-heteroaryl), or a pharmaceutically acceptable salt or stereoisomer thereof.

[0085] In some embodiments, the compound has formula (Ia): [ka] (In the formula, B 1 and B 2 each of X is independently a purinyl nucleobase or a pyrimidinyl nucleobase; 1 and X 2 each independently is O or S; and Y 1 and Y 2 each independently represents O, S, or NR 5 and L 1 and L 2Each of C1 to C2 is independently absent or 20 Alkyl or C1-C 20 heteroalkyl, wherein each of the alkyl and heteroalkyl is optionally R 6 is substituted with R 1 and R 2 each independently represents hydrogen, halo, -CN, C1-C 20 Alkyl, or OR 7 and R 3 and R 4 each independently represents hydrogen, C1 to C 20 Alkyl, C1-C 20 heteroalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein the alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each optionally have 1 to 5 R 8 is substituted with R 5 is hydrogen or C1-C 20 alkyl, and R 6 Halo, -CN, C1~C 20 Alkyl, OR 7 , oxo, cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein each alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally joined to 1 to 5 R 9 is substituted with R 7 is hydrogen, C1 to C 20 alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein each alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl optionally has 1 to 5 R 9 is substituted with R 8 are each independently C1 to C 20 Alkyl, C1-C 20 Heteroalkyl, C(O)-C1-C 20 Alkyl, OC(O)-C1-C 20 Alkyl, C(O)O-C1-C 20 Alkyl, OC(O)O-C1-C 20 Alkyl, C(O)N(R 5 )-C1~C20 Alkyl, N(R 5 )C(O)-C1~C 20 Alkyl, OC(O)N(R 5 )-C1~C 20 Alkyl, O-aryl, O-heteroaryl, C(O)-aryl, C(O)-heteroaryl, OC(O)-aryl, OC(O)-heteroaryl, C(O)O-aryl, C(O)O-heteroaryl, C(O)N(R 5 )-aryl, C(O)N(R 5 )-heteroaryl, N(R 5 )C(O)-aryl, N(R 5 )C(O)-heteroaryl, where alkyl, heteroalkyl, aryl, or heteroaryl each have 1 to 5 R 9 is substituted with R 9 are each independently C1 to C 20 Alkyl, O-C1-C 20 Alkyl, C1-C 20 heteroalkyl, halo, -CN, OH, oxo, aryl, heteroaryl, O-aryl, or O-heteroaryl, where alkyl, heteroalkyl, aryl) or a pharmaceutically acceptable salt or stereoisomer thereof.

[0086] In some embodiments, the compound has formula (Ib), (Ic), (Id), or (Ie): [ka] (In the formula, B 1 , B 2 , X 1 , X 2 , Y 1 , Y 2 , L 1 , L 2 , R 1 , R 2 , R 3 , R 4 and each of these subvariables is as previously described), or a pharmaceutically acceptable salt thereof.

[0087] In some embodiments, B 1 or B 2 At least one of B is a purinyl nucleobase. 1 or B 2 Each of B is independently a purinyl nucleobase. 1 is a purinyl nucleobase. In some embodiments, B 2 is a pyrimidinyl nucleobase. In some embodiments, B 1 is a purinyl nucleobase, and B 2 is a pyrimidinyl nucleobase.

[0088] In some embodiments, B 1 or B 2 Each of B is selected from a natural nucleobase or a modified nucleobase. 1 or B 2 Each of B is selected from adenosinyl, guanosinyl, cytosinyl, thyminyl, uracilyl, 5'-methylcytosinyl, 5'-fluorouracilyl, 5'-propynyluracilyl, and 7-deazaadenosinyl. 1 or B 2 Each of the [ka] is selected from: [ka] indicates the attachment of the nucleobase to the ribose ring.

[0089] In some embodiments, B 1 or B 2 one of which is selected from natural nucleobases, and B 1 or B 2 The other of B is a modified nucleobase. 1 or B 2is adenosinyl, guanosinyl, thyminyl, cytosinyl, or uracilyl, and B 1 or B 2 The other is 5'-methylcytosinyl, 5'-fluorouracil, 5'-propynyluracil, or 7-deazaadenosinyl.

[0090] In some embodiments, B 1 is adenosinyl or guanosinyl. In some embodiments, B 2 is cytosinyl, thyminyl, or uracilyl. 1 is adenosinyl or guanosinyl, and B 2 is cytosinyl, thyminyl, or uracilyl. 1 and B 2 Each of B is independently uracilyl. 1 and B 2 Each of is independently adenosinyl.

[0091] In some embodiments, R 1 and R 2 each independently is hydrogen, halo, or OR 7 In some embodiments, R 1 and R 2 Each of R is independently halo (e.g., fluoro). 1 and R 2 Each of 7 isn't it.

[0092] In some embodiments, X 1 is O. In some embodiments, X 2 is O. In some embodiments, X 1 and X 2 Each of is independently O.

[0093] In some embodiments, Y 1is O or S. In some embodiments, Y 2 is O or S. In some embodiments, Y 1 and Y 2 Each of Y is independently O or S. In some embodiments, Y 1 or Y 2 One of the two is O and the other is Y 1 or Y 2 and the other is S. In some embodiments, Y 1 or Y 2 Each of Y is independently S. In some embodiments, Y 1 or Y 2 Each of is independently O.

[0094] In some embodiments, L 1 is C1-C6 alkyl (e.g., CH2). In some embodiments, L 2 is C1-C6 alkyl (e.g., CH2). In some embodiments, L 1 and L 2 Each of is independently C1-C6 alkyl (eg, CH2).

[0095] In some embodiments, R 3 is hydrogen, aryl, or heteroaryl, where aryl and heteroaryl are optionally joined by 1 to 5 R 8 In some embodiments, R 3 is aryl or heteroaryl, each of which optionally has 1 to 5 R 8 In some embodiments, R 3 is one R 8 is a phenyl substituted with

[0096] In some embodiments, R 4 are independently hydrogen, aryl, or heteroaryl, wherein the aryl and heteroaryl are optionally joined by 1 to 5 R 8 In some embodiments, R4 is aryl or heteroaryl, each of which optionally has 1 to 5 R 8 In some embodiments, R 4 is one R 8 is a phenyl substituted with

[0097] In some embodiments, R 3 and R 4 is independently hydrogen, aryl, or heteroaryl, wherein the aryl and heteroaryl are optionally joined by 1 to 5 R 8 In some embodiments, R 3 is aryl or heteroaryl, each of which optionally has 1 to 5 R 8 is substituted with R 4 is hydrogen. In some embodiments, R 3 is one R 8 is phenyl substituted with R 4 is hydrogen. In some embodiments, R 3 and R 4 Each of the groups independently represents one R 8 is a phenyl substituted with

[0098] In some embodiments, Y 1 and Y 2 Each of is O and R 3 and R 4 Each of Y is independently hydrogen. 2 is O and R 4 is hydrogen. In some embodiments, Y 1 and Y 2 each independently is S, and R 3 and R 4 Each of the groups independently represents one R 8 In some embodiments, Y 1 is S and R 3 is one R 8 is replaced by .

[0099] In some embodiments, R 8 are each independently C1 to C 20 Alkyl (e.g., C1-C6 alkyl), C1-C 20 Heteroalkyl, C(O)-C1-C 20 Alkyl, OC(O)-C1-C 20 Alkyl, OC(O)O-C1-C 20 Alkyl, OC(O)N(R 5 )-C1~C 20 Alkyl, O-aryl, C(O)-aryl, OC(O)-aryl, or C(O)N(R 5 )-aryl, wherein alkyl, heteroalkyl, aryl, and heteroaryl are each optionally joined by one or more R 9 has been replaced by

[0100] In some embodiments, R 8 is any 1 to 5 R 9 (For example, one R 9 ) is O-C(O)-aryl substituted by

[0101] In some embodiments, R 9 is O-C1~C 12 alkyl (e.g., O-CH(CH)CH). In some embodiments, R 9 is O-C1~C 10 alkyl (e.g., O-CH(CH)CH). In some embodiments, R 9 is O—C1-C8 alkyl (e.g., O—CH2(CH2)6CH3). In some embodiments, R 9 is O-C1-C6 alkyl (for example, O-CH2(CH2)4CH3).

[0102] In some embodiments, the compound has the formula (If): [ka] (In the formula, B 1 and B 2each of X is independently a purinyl nucleobase or a pyrimidinyl nucleobase; 1 and X 2 each independently is O or S; and Y 1 and Y 2 each independently represents O, S, or NR 5 and L 1 and L 2 each independently is absent, C1-C6 alkyl, or C1-C6 heteroalkyl, wherein each of the C1-C6 alkyl and C1-C6 heteroalkyl is optionally selected from the group consisting of R 6 is substituted with R 1 and R 2 each independently is halo; R 3 and R 4 each independently represents hydrogen, C1 to C 20 alkyl, C1-C6 heteroalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl, where C1-C 20 Alkyl, C1-C6 heteroalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each optionally have 1 to 5 R 8 is substituted with R 5 is hydrogen or C1-C 20 alkyl, and R 6 is halo, -CN, C1~C 20 Alkyl, OR 7 , oxo, cycloalkyl, heterocyclyl, aryl, or heteroaryl, where C1-C 20 Each alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl may optionally have 1 to 5 R 9 is substituted with R 7 is hydrogen, C1 to C 20 alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl, where C1-C 20 Each alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl may optionally have 1 to 5 R 9 is substituted with R 8 are each independently C1 to C 20alkyl, C(O)-aryl, C(O)-heteroaryl, OC(O)-aryl, or OC(O)-heteroaryl, where C1-C 20 Alkyl, C(O)-aryl, C(O)-heteroaryl, OC(O)-aryl, or OC(O)-heteroaryl may each optionally be one to five R 9 is replaced by R 9 are each independently C1 to C 20 Alkyl, halo, -CN, OH, O-C1~C 20 Alkyl, O-C1-C 20 or a pharmaceutically acceptable salt or stereoisomer thereof.

[0103] In some embodiments, the compound has the formula (Ig): [ka] (In the formula, B 1 and B 2 each of X is independently a purinyl nucleobase or a pyrimidinyl nucleobase; 1 and X 2 each independently is O; and Y 1 and Y 2 each independently is O or S, and L 1 and L 2 are independently absent or C1-C6 alkyl, and R 1 and R 2 each independently is halo or OH, and R 3 and R 4 each independently represents hydrogen, or optionally 1 to 5 R 8 aryl substituted with R 8 each independently represents 1 to 5 R 9 and R is an OC(O)-aryl substituted by 9 are each independently O-C1 to C 20 or a pharmaceutically acceptable salt or stereoisomer thereof.

[0104] In some embodiments, the compound is selected from the compounds shown in Table 1. [Table 1] TIFF0007749629000024.tif230167TIFF0007749629000025.tif217166TIFF0007749629000026.tif238166TIFF0007749629000027.tif225163TIFF0007749629000028.tif231164, where X is any pharmaceutically acceptable counterion, e.g., lithium, sodium, potassium, calcium, magnesium, aluminum, ammonium, ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, etc. (See, e.g., Berge et al., supra.) In some embodiments, the compounds of Table 1 are not salts (e.g., are free acids or free bases).

[0105] In some embodiments, the compound is a compound shown in Table 2. [Table 2] TIFF0007749629000030.tif232168TIFF0007749629000031.tif203167TIFF0007749629000032.tif2371 66TIFF0007749629000033.tif217167TIFF0007749629000034.tif200165TIFF0007749629000035.tif22 4167TIFF0007749629000036.tif211169TIFF0007749629000037.tif219167TIFF0007749629000038.tif231169TIFF0007749629000039.tif224167TIFF0007749629000040.tif49168 or a pharmaceutically acceptable salt thereof.

[0106] In one embodiment, the compounds described herein are in the form of a pharmaceutically acceptable salt. Exemplary salts, such as ammonium salts, are described herein. In some embodiments, the compounds are monosalts. In some embodiments, the compounds are di-salts. In some embodiments, the compounds described herein (e.g., compounds in Table 1 or Table 2) are not salts (e.g., are free acids or free bases).

[0107] The compounds of formula (I) or formula (Ia) are small molecule nucleic acid complex (cyclic dinucleotide) compounds that combine both antiviral and immunomodulatory activity, the latter activity mediating controlled apoptosis of virus-infected hepatocytes via stimulation of the innate immune response, similar to that achieved by IFN-α therapy in patients suffering from viral infections.

[0108] Without wishing to be bound by theory, the mechanism of action of the compounds of Formula (I) or Formula (Ia) involves the host immune stimulatory activity of the compounds, which can induce endogenous IFN through activation of PRRs such as RIG-I, NOD2, and STING. As previously described, activation can occur by binding of the compounds of Formula (I) to the nucleotide-binding domain of PRRs (e.g., STING), which can result in the induction of PRR expression (e.g., STING expression).

[0109] The compounds provided herein may contain one or more asymmetric centers and thus occur as racemates and racemic mixtures, single enantiomers, individual diastereomers, and diastereomeric mixtures.All such isomeric forms of these compounds are expressly included within the scope of the present invention.When a compound is named or depicted by structure without specifying stereochemistry, and has one or more chiral centers, it is understood to represent all possible stereoisomers of the compound unless otherwise stated.The compounds provided herein may also contain restrictions resulting from the presence of bonds (e.g., carbon-carbon bonds, phosphorus-oxygen bonds, or phosphorus-sulfur bonds) or substituents that can restrict bond rotation, such as rings or double bonds.

[0110] In some embodiments, the methods described herein include administering a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, the methods described herein include administering a compound of Formula (Ia) or a pharmaceutically acceptable salt thereof. In some embodiments, the compound of Formula (I) includes an isomer of the compound of Formula (I) (e.g., an Rp-isomer or an Sp-isomer) or a mixture of isomers of the compound of Formula (I) (e.g., an Rp-isomer or an Sp-isomer). In some embodiments, the compound of Formula (I) includes an isomer of the compound of Formula (Ia) (e.g., an Rp-isomer or an Sp-isomer) or a mixture of isomers of the compound of Formula (Ia) (e.g., an Rp-isomer or an Sp-isomer).

[0111] How to use The present disclosure relates to methods of inducing expression of a PRR (e.g., STING) in a subject through administration of a compound of formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, the subject may be suffering from a condition described below, e.g., a proliferative disease, e.g., cancer.

[0112] It has been reported that many patients suffering from advanced solid tumors exhibit a spontaneous T cell-mediated inflammatory tumor microenvironment that predicts prognosis and clinical response to immunotherapy. Recent findings suggest that the cytosolic DNA-sensing STING pathway is a critical innate immune sensing mechanism that drives type I IFN production in the tumor context. Knowledge of this pathway is guiding the further development of novel immunotherapeutic strategies.

[0113] It has been reported that the presence of activated CD8+ T cells within the tumor microenvironment in early-stage colorectal cancer carries significant positive prognostic significance. Patients with other solid tumor histologies also appear to have spontaneous T cell infiltrates, which may have similar positive prognostic value. These include breast cancer, renal cell carcinoma, melanoma, ovarian cancer, and gastrointestinal tumors. T cell infiltrates are thought to contain tumor antigen-specific T cells that are spontaneously activated in response to the growing tumor, likely through immune surveillance. This attempted host immune response, even if it does not completely eliminate the tumor, is thought to slow tumor progression and thus result in improved clinical outcomes. Furthermore, innate immune mechanisms can generate adaptive T cell responses to tumor antigens even in the absence of exogenous infection. In this regard, human cancer gene expression profiling studies have revealed a relationship between type I IFN signatures, T cell infiltration, and clinical outcomes. Therefore, the innate immune sensing pathway that triggers type I IFN production may represent an important intermediate mechanistic step. Gene expression profiling of melanoma revealed two major tumor microenvironment subsets that either exhibit transcriptional signatures indicative of T cell infiltration or lack thereof. Indeed, the CD8+ T cells, macrophages, and some B cells and plasma cells in these lesions in melanoma metastases are phenotypically similar to those described in early colon cancer and other tumors where activated T cells have been associated with a favorable prognosis. CD8+ T cells were required for the upregulation of all immune factors within the tumor microenvironment. Studies have shown that IFN production is necessary for optimal T cell priming against tumor antigens. Many PRRs, including STING, induce IFN-β production by host DCs in response to tumor growth in vivo. STING is an adaptor protein activated by cyclic dinucleotides generated by cyclic GMP-AMP synthase (cGAS), which then directly activates cytosolic DNA.In the presence of these cyclic dinucleotides and / or DNA, STING translocates from the endoplasmic reticulum to various perinuclear compartments; for example, palmitoylation of STING in the Golgi apparatus has been shown to be essential for STING activation (Mukai, K. et al. (2016) Nat Commun doi:10.1038 / ncomms11932).

[0114] Activated STING forms aggregates and activates TBK1, which then phosphorylates interferon regulatory factor 3 (IRF3), which directly contributes to type I IFN gene transcription. This pathway has been implicated in DNA virus sensing and in selected autoimmune models. Furthermore, activating STING mutations have recently been identified in human patients suffering from vasculitis / pulmonary inflammatory syndrome, characterized by increased type I IFN production. Mechanistic studies using mouse transplantable tumor models have revealed that STING knockout and IRF3 knockout mice exhibit defective spontaneous T cell priming against tumor antigens in vivo, resulting in abrogated rejection of immunogenic tumors. Similarly, tumor-derived DNA was found in the cytosol of a major population of tumor-infiltrating DCs, which was associated with STING pathway activation and IFN-β production. Therefore, the host STING pathway appears to be a critical innate immune sensing pathway that detects the presence of tumors and drives DC activation and subsequent T cell priming against tumor-associated antigens in vivo. The functional role of the STING pathway in vivo has also been reported in other mouse-tumor systems. An inducible glioma model was shown to result in the induction of a type I IFN gene signature as part of the host response. This induction was substantially reduced in STING knockout mice, resulting in more aggressive tumor growth and shorter mouse survival. Exogenous delivery of cyclic dinucleotides as STING agonists exerted therapeutic effects in vivo. The STING pathway was also confirmed in B16.OVA and EL4.OVA models in response to cryoablation. Interestingly, host STING is also required for maximal production of anti-DNA antibodies, so the mechanisms involved parallel those observed in the Bm12 mouse model of lupus. Thus, the antitumor immune response induced in part by tumor DNA overlaps with mechanisms involved in autoimmunity driven by extracellular DNA. The role of STING has also been explored in an inducible colon cancer model. The ability of individual patient cancers to support STING pathway activation appears to be linked to the spontaneous development of a T cell-inflammatory tumor microenvironment.Because this phenotype correlates with improved prognosis in patients with early-stage cancer and clinical response to immunotherapy in the metastatic setting, impaired STING activation may therefore represent an early functional block and thus may have prognostic / predictive value as a biomarker in its own right. Second, strategies that activate or mimic the output of the host STING pathway should have immunotherapeutic potential in the clinic. To the extent that non-T cell inflammatory tumors appear to lack evidence of a type I IFN transcriptional signature, strategies that promote robust innate signaling via APCs in the tumor microenvironment may facilitate improved cross-priming of tumor antigen-specific CD8+ T cells and subsequently enhance chemokine production for tumor-regressing activity.

[0115] Cancer treatment Recognition of nucleic acid ligands by PRRs such as cGAS, RIG-I, and STING stimulates the production of type I interferons (e.g., IFN-α or IFN-β), triggering a series of downstream signaling events that can lead to apoptosis in susceptible cells. Recently, a link between the induction of PRR expression and numerous cancers has been discovered. For example, RIG-I expression has been shown to be significantly downregulated in hepatocellular carcinoma, and patients with low RIG-I expression in tumors had shorter survival times and poorer responses to IFN-α therapy (Hou, J. et al., Cancer Cell (2014) 25:49-63). It has been suggested that the level of RIG-I expression may be useful as a biomarker for predicting prognosis and response to immunotherapy. In other cases, induction of RIG-I expression has been shown to induce immunogenic cell death in pancreatic, prostate, breast, skin, and lung cancer cells (Duewell, P. et al, Cell Death Differ (2014) 21:1825-1837; Besch, R. et al, J Clin Invest (2009) 119:2399-2411; Kaneda, Y. Oncoimmunology (2013) 2:e23566; Li, XY et al, Mol Cell Oncol (2014) 1:e968016), highlighting new approaches in immune-mediated cancer therapy.

[0116] STING is recognized as a key adaptor protein in the cGAS-STING-IFN cascade, but it has also been reported to be a sensor for DNA. A role for STING in stimulating innate immunity in response to cancer has also been identified. Recent studies have revealed the presence of tumor-derived DNA in the cytosol of certain antigen-presenting cells, such as tumor-infiltrating dendritic cells, likely arising through tumor cell stress or cell death. This tumor-derived DNA is known to trigger the production of cyclic nucleotides, which have been shown to activate STING, leading to cGAS activation and consequent production of the associated type 1 interferons (Woo, SR et al, Immunity (2014) 41:830-842). STING stimulation and the resulting downstream signaling pathways also appear to contribute to the recruitment of effector T cells to the inflamed tumor microenvironment (Woo, SR Trends in Immunol (2015) 36:250-256). STING activation in the tumor microenvironment can induce adaptive immune responses that result in antitumor activity. Thus, in this setting, tumors that are STING-deficient, those described herein may still have anti-tumor activity through activation of antigen-presenting cells and dendritic cells (APCs and DCs) and induction of adaptive immune responses.

[0117] In some embodiments, a method for inducing expression of a PRR (e.g., a PRR described herein) comprises administering a compound of Formula (I) or a pharmaceutically acceptable salt thereof to a subject suffering from cancer. In some embodiments, a method for inducing expression of a PRR (e.g., a PRR described herein) comprises administering a compound of Formula (Ia) or a pharmaceutically acceptable salt thereof to a subject suffering from cancer. In some embodiments, a method for inducing STING expression disclosed herein comprises administering a compound of Formula (I) or a pharmaceutically acceptable salt thereof to a subject suffering from cancer. In some embodiments, a method for inducing STING expression disclosed herein comprises administering a compound of Formula (Ia) or a pharmaceutically acceptable salt thereof to a subject suffering from cancer. In some embodiments, a method for inducing RIG-I expression disclosed herein comprises administering a compound of Formula (I) or a pharmaceutically acceptable salt thereof to a subject suffering from cancer. In some embodiments, a method for inducing RIG-I expression disclosed herein comprises administering a compound of Formula (Ia) or a pharmaceutically acceptable salt thereof to a subject suffering from cancer. In some embodiments, the methods for inducing NOD2 expression disclosed herein comprise administering a compound of Formula (I) or a pharmaceutically acceptable salt thereof to a subject suffering from cancer. In some embodiments, the methods for inducing NOD2 expression disclosed herein comprise administering a compound of Formula (Ia) or a pharmaceutically acceptable salt thereof to a subject suffering from cancer. In some embodiments, the cancer is selected from cancer of the breast, bone, brain, cervix, colon, gastrointestinal tract, eye, gallbladder, lymph node, blood, lung, liver, skin, oral cavity, prostate, ovary, penis, pancreas, uterus, testicle, stomach, thymus, thyroid, or other body parts. In some embodiments, the cancer comprises a solid tumor (e.g., carcinoma, sarcoma, or lymphoma). In some embodiments, the cancer is hepatocellular carcinoma or other liver cancer. In some embodiments, the cancer is leukemia or other blood cancer.In some embodiments, the cancer comprises breast cancer, renal cell carcinoma, colon cancer, melanoma, ovarian cancer, head and neck squamous cell carcinoma, pancreatic cancer, prostate cancer, lung cancer, brain cancer, thyroid cancer, kidney cancer, testicular cancer, gastric cancer, urothelial cancer, skin cancer, cervical cancer, endometrial cancer, liver cancer, lung cancer, lymphoma or gastrointestinal stromal cancer, and solid tumors. In some embodiments, the cancer cells (e.g., tumor cells) comprise a specific cancer-associated antigen that induces a T cell-mediated anti-tumor response.

[0118] In some embodiments, the methods disclosed herein for inducing expression of a PRR (e.g., STING, RIG-I, MDA5, LGP2) in a subject suffering from cancer result in increased PRR expression (e.g., STING expression). In some embodiments, expression of a PRR (e.g., STING) is induced by about 1.1, about 1.2, about 1.3, about 1.4, about 1.5, about 1.6, about 1.7, about 1.8, about 1.9, about 2, about 2.5, about 3, about 4, about 5, about 7.5, about 10, about 15, about 20, about 25, about 30, about 40, about 50, about 75, about 100, about 150, about 200, about 250, about 500, about 1000, about 1500, about 2500, about 5000, about 10,000, or more factors. In some embodiments, induction of expression of a PRR (e.g., STING) occurs within about 5 minutes of administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, induction of expression of a PRR (e.g., STING) occurs within about 5 minutes of administration of a compound of Formula (Ia) or a pharmaceutically acceptable salt thereof. In some embodiments, induction of expression of a PRR (e.g., STING) occurs within about 5 minutes of administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, induction of expression of a PRR (e.g., STING) occurs within about 10 minutes, about 15 minutes, about 20 minutes, about 25 minutes, about 30 minutes, about 45 minutes, about 1 hour, about 1.5 hours, 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 10 hours, about 12 hours or more after administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, induction of expression of a PRR (e.g., STING) occurs within about 10 minutes, about 15 minutes, about 20 minutes, about 25 minutes, about 30 minutes, about 45 minutes, about 1 hour, about 1.5 hours, 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 10 hours, about 12 hours or more after administration of a compound of Formula (Ia) or a pharmaceutically acceptable salt thereof.It is recognized that activation of STING by compounds can lead to the induction of expression of other PRRs, such as RIG-I, MDA5, and NOD2, which can further amplify IFN production in the tumor microenvironment and stimulate T cells for enhanced antitumor activity.

[0119] In some embodiments, the methods disclosed herein for inducing PRR (e.g., STING) expression in a subject suffering from cancer result in increased PRR expression (e.g., STING expression). In some embodiments, PRR (e.g., STING) expression is induced by about 1.1, about 1.2, about 1.3, about 1.4, about 1.5, about 1.6, about 1.7, about 1.8, about 1.9, about 2, about 2.5, about 3, about 4, about 5, about 7.5, about 10, about 15, about 20, about 25, about 30, about 40, about 50, about 75, about 100, about 150, about 200, about 250, about 500, about 1000, about 1500, about 2500, about 5000, about 10,000, or more factors. In some embodiments, induction of PRR (e.g., STING) expression occurs within about 5 minutes of administration of a compound of Formula (I) or a pharmaceutically acceptable salt or stereoisomer thereof. In some embodiments, induction of PRR (e.g., STING) expression occurs within about 5 minutes of administration of a compound of Formula (Ia) or a pharmaceutically acceptable salt or stereoisomer thereof. In some embodiments, induction of PRR (e.g., STING) expression occurs within about 10 minutes, about 15 minutes, about 20 minutes, about 25 minutes, about 30 minutes, about 45 minutes, about 1 hour, about 1.5 hours, 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 10 hours, about 12 hours or more after administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, induction of expression of a PRR (e.g., STING) occurs within about 10 minutes, about 15 minutes, about 20 minutes, about 25 minutes, about 30 minutes, about 45 minutes, about 1 hour, about 1.5 hours, 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 10 hours, about 12 hours or more after administration of a compound of Formula (Ia) or a pharmaceutically acceptable salt thereof.

[0120] Pharmaceutical Composition The disclosure features a method for inducing PRR (e.g., STING) expression in a subject, the method including administering a compound of Formula (I) or Formula (1-a), or a pharmaceutically acceptable salt thereof.

[0121] Although it is possible to administer a compound of the present disclosure (e.g., a compound of Formula (I)) alone, it is preferable to administer the compound as a pharmaceutical composition or formulation, in which the compound is combined with one or more pharmaceutically acceptable diluents, excipients, or carriers. Compounds according to the present disclosure can be formulated for administration in any convenient manner for use in human or veterinary medicine. In certain embodiments, the compound included in the pharmaceutical preparation can be active itself or, for example, a prodrug that can be converted to an active compound in a physiological setting. Regardless of the selected route of administration, the compounds of the present disclosure, which can be used in a suitable hydrated form, and / or pharmaceutical compositions of the present disclosure are formulated into pharmaceutically acceptable dosage forms as described below or by other conventional methods known to those skilled in the art.

[0122] The amount and concentration of a compound of the present disclosure (e.g., a compound of Formula (I)) in a pharmaceutical composition, and the amount of the pharmaceutical composition administered to a subject, can be selected based on clinically relevant factors, such as the subject's medically relevant characteristics (e.g., age, weight, sex, other medical conditions, etc.), the solubility of the compound in the pharmaceutical composition, the potency and activity of the compound, and the mode of administration of the pharmaceutical composition. For further information on routes of administration and dosage regimens, the reader is referred to Chapter 25.3 in Volume 5 of Comprehensive Medicinal Chemistry (Corwin Hansch; Chairman of the Editorial Board), Pergamon Press, 1990.

[0123] Accordingly, another aspect of the present disclosure provides pharmaceutically acceptable compositions comprising a therapeutically or prophylactically effective amount of a compound described herein (e.g., a compound of Formula (I)) formulated together with one or more pharmaceutically acceptable carriers (excipients) and / or diluents. As described in detail below, the pharmaceutical compositions of the present disclosure may be specially formulated for administration in solid or liquid form, including oral administration, intratumoral administration, parenteral administration, e.g., suitable for subcutaneous, intramuscular, intraperitoneal, or intravenous injection as a sterile solution or suspension. However, in certain embodiments, the compounds of the present invention can simply be dissolved or suspended in sterile water. In certain embodiments, the pharmaceutical preparations are non-pyrogenic, i.e., do not elevate a patient's body temperature.

[0124] As used herein, the phrases "systemic administration," "administered systemically," "peripheral administration," and "administered peripherally" refer to administration of a compound other than directly into the central nervous system, whereby the compound enters the patient's system and is therefore subject to metabolism and other similar processes, for example, subcutaneous administration.

[0125] The phrase "pharmaceutically acceptable" is employed herein to refer to those compounds, materials, compositions, and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0126] As used herein, the phrase "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, stabilizer, excipient, solvent, or encapsulating material, that is involved in carrying or transporting an antagonist of the present invention from one organ or part of the body to another. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not harmful to the patient. Some examples of materials that can serve as pharmaceutically acceptable carriers include, but are not limited to, (1) sugars, such as lactose, glucose, and sucrose. (2) starches such as corn starch and potato starch, (3) cellulose and its derivatives such as sodium carboxymethylcellulose, ethyl cellulose, and cellulose acetate, (4) powdered tragacanth, (5) malt, (6) gelatin, (7) talc, (8) excipients such as cocoa butter and suppository wax, (9) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil, (10) glycols such as propylene glycol, (11) polyols such as glycerin, sorbitol, mannitol, and polyethylene glycol, (12) esters such as ethyl oleate and ethyl laurate, (13) agar, and (14) buffers such as magnesium hydroxide and aluminum hydroxide. (15) alginic acid, (16) ascorbic acid, (17) pyrogen-free water, (18) isotonic saline, (19) Ringer's solution, (20) ethyl alcohol, (21) phosphate buffer, (22) cyclodextrins such as Captisol®, and (23) other non-toxic compatible substances such as antioxidants and antimicrobial agents employed in pharmaceutical formulations.

[0127] As mentioned above, certain embodiments of the compounds described herein may contain a basic functional group, such as an amine, and therefore can form pharmaceutically acceptable salts with pharmaceutically acceptable acids. In this respect, the term "pharmaceutically acceptable salts" refers to the relatively non-toxic inorganic and organic acid addition salts of the compounds of the present disclosure. These salts can be prepared in situ during the final isolation and purification of the compounds of the present disclosure, or by separately reacting the purified compounds of the present disclosure in their free base form with a suitable organic or inorganic acid and isolating the salt thus formed. Representative salts include hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate, mesylate, glucoheptonate, lactobionate, and lauryl sulfonate salts, and the like (see, e.g., Berge et al. (1977) "Pharmaceutical Salts", J. Pharm. Sci. 66:1-19).

[0128] In other cases, compounds of the present disclosure may contain one or more acidic functional groups and thus can form pharmaceutically acceptable salts with pharmaceutically acceptable bases. In these cases, the term "pharmaceutically acceptable salts" refers to relatively non-toxic inorganic and organic base addition salts of compounds of the present disclosure (e.g., compounds of Formula (I)). These salts can also be prepared in situ during the final isolation and purification of the compounds, or by separately reacting the purified compounds of the present disclosure in their free acid form with a suitable base, such as a hydroxide, carbonate, or bicarbonate of a pharmaceutically acceptable metal cation, with ammonia, or with a pharmaceutically acceptable organic primary, secondary, or tertiary amine. Representative alkali or alkaline earth salts include lithium, sodium, potassium, calcium, magnesium, and aluminum salts, and the like. Representative organic amines useful for forming base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, and the like (see, for example, Berge et al., supra).

[0129] Wetting agents, emulsifiers, and lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweeteners, flavorings and fragrances, preservatives, and antioxidants can also be present in the composition. Examples of pharmaceutically acceptable antioxidants include (1) water-soluble antioxidants such as ascorbic acid, cysteine ​​hydrochloride, sodium bisulfate, sodium metabisulfite, and sodium sulfite; (2) oil-soluble antioxidants such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, and α-tocopherol; and (3) metal chelating agents such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, and phosphoric acid.

[0130] Pharmaceutically acceptable carriers, as well as wetting agents, emulsifiers, lubricants, colorants, release agents, coating agents, sweeteners, flavoring agents, perfumes, preservatives, antioxidants, and other additional ingredients may be present in amounts of about 0.001% to 99% of the compositions described herein. For example, the pharmaceutically acceptable carrier, as well as wetting agents, emulsifiers, lubricants, colorants, release agents, coating agents, sweeteners, flavoring agents, perfumes, preservatives, antioxidants, and other additional ingredients may be present from about 0.005%, about 0.01%, about 0.05%, about 0.1%, about 0.25%, about 0.5%, about 0.75%, about 1%, about 1.5%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 85%, about 90%, about 95%, or about 99% of the compositions described herein.

[0131] The pharmaceutical compositions of the present disclosure may be in a form suitable for oral administration, for example, a liquid or solid oral dosage form. In some embodiments, liquid dosage forms include suspensions, solutions, electuaries, emulsions, beverages, elixirs, or syrups. In some embodiments, solid dosage forms include capsules, tablets, powders, dragees, or powders. The pharmaceutical compositions may be in unit dosage forms suitable for single administration of precise dosages. In addition to a compound described herein (e.g., a compound of Formula (I)) or a pharmaceutically acceptable salt thereof, the pharmaceutical compositions may include pharmaceutically acceptable carriers such as stabilizers (e.g., binders, e.g., polymers, e.g., suspending agents, diluents, binders, and lubricants).

[0132] In some embodiments, the compositions described herein comprise a liquid dosage form for oral administration, such as a solution or suspension. In other embodiments, the compositions described herein comprise a solid dosage form for oral administration that can be directly compressed into a tablet. Additionally, the tablet may contain other medicinal or pharmaceutical agents, carriers, and / or adjuvants. Exemplary pharmaceutical compositions include, for example, compressed tablets (e.g., direct compression tablets) containing a compound of the present disclosure (e.g., a compound of Formula (I)) or a pharmaceutically acceptable salt thereof.

[0133] The formulations of the present disclosure include those suitable for parenteral administration. The formulations can be conveniently provided in unit dosage form and can be prepared by any method well known in the pharmaceutical arts. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will vary depending on the host being treated and the particular mode of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will generally be the amount of compound that produces a therapeutic effect. Generally, out of 100 percent, this amount will range from about 1 percent to about 99 percent of the active ingredient, preferably from about 5 percent to about 70 percent, and most preferably from about 10 percent to about 30 percent. Pharmaceutical compositions of the present disclosure suitable for parenteral administration comprise a compound of the present disclosure in combination with one or more pharmaceutically acceptable sterile, isotonic aqueous or non-aqueous solvents, dispersions, suspensions or emulsions, which may contain antioxidants, buffers, bacteriostats, solutes that render the formulation isotonic with the blood of the intended recipient, or suspending or thickening agents, or sterile powders that can be reconstituted into sterile injectable solutions or dispersions immediately before use.

[0134] Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions of the present disclosure include water, ethanol, polyols (glycerol, propylene glycol, polyethylene glycol, etc.) and suitable mixtures thereof, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate.Proper fluidity can be maintained, for example, by using coating materials such as lecithin, by maintaining the required particle size in the case of dispersions, and by using surfactants.

[0135] These compositions may contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of microbial action can be ensured by including various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol sorbic acid, etc. It may also be desirable to include isotonic agents such as sugars and sodium chloride in the composition. In addition, prolonged absorption of injectable pharmaceutical forms can be achieved by including agents that delay absorption, such as aluminum monostearate and gelatin.

[0136] In some cases, in order to prolong the effect of the compounds of the present disclosure (e.g., compounds of formula (I)), it may be desirable to delay the absorption of the drug from subcutaneous, intraperitoneal, or intramuscular injection. This can be achieved by using a liquid suspension of crystalline or amorphous material that is poorly water-soluble. The absorption rate of the drug then depends on its dissolution rate, which in turn may depend on the size and crystalline form of the crystal. Alternatively, delayed absorption of the compounds of the present disclosure in parenteral administration form can be achieved by dissolving or suspending the compound in an oil vehicle.

[0137] In some embodiments, it may be advantageous to administer the compound of the present disclosure (e.g., compound of formula (I)) in a sustained manner.It is recognized that any formulation that provides sustained absorption characteristics can be used.In certain embodiments, sustained absorption can be achieved by combining the compound of the present disclosure with other pharmaceutically acceptable ingredients, diluents, or carriers that delay the release characteristics of the compound of the present disclosure into the systemic circulation.

[0138] Administration route The compounds and compositions used in the methods described herein can be administered to a subject in a variety of forms depending on the selected route of administration, as will be understood by those skilled in the art. Exemplary routes of administration for the compositions used in the methods described herein include topical, enteral, or parenteral application. Topical application includes transdermal, inhalation, enemas, eye drops, ear drops, and application via internal mucous membranes. Enteral application includes oral, rectal, vaginal, and tube feeding. Parenteral administration includes intravenous, intraarterial, intravesical, intraorbital, intracardiac, intradermal, intratracheal, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, intraosseous, intraperitoneal, subcutaneous, intramuscular, transepithelial, intranasal, intrapulmonary, intrathecal, intrarectal, and topical modes of administration. Parenteral administration can be by continuous infusion over a selected period of time. In certain embodiments of the present disclosure, the compositions described herein comprising a compound of Formula (I) are administered orally. In certain embodiments of the present disclosure, the compositions described herein comprising a compound of Formula (Ia) are administered orally. In other embodiments of the present disclosure, the compositions described herein comprising a compound of Formula (I) are administered parenterally (e.g., intraperitoneally). It is recognized that for the treatment of solid tumors, direct injection of the compound into the tumor may also be performed (e.g., intratumoral administration). In other embodiments of the present disclosure, the compositions described herein comprising a compound of Formula (Ia) are administered parenterally (e.g., intraperitoneally). It is recognized that for the treatment of solid tumors, direct injection of the compound into the tumor may also be performed (e.g., intratumoral administration).

[0139] For intravenous, intraperitoneal, or intrathecal delivery or direct injection (e.g., intratumor), the composition must be sterile and fluid to the extent that the composition can be delivered by syringe. In addition to water, the carrier can be an isotonic buffered saline solution, ethanol, polyol (e.g., glycerol, propylene glycol, liquid polyethylene glycol, and the like), and suitable mixtures thereof. Proper fluidity can be maintained, for example, by the use of coating materials such as lecithin, by maintaining the required particle size in the case of dispersions, and by the use of surfactants. In many cases, it is preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol or sorbitol, and sodium chloride in the composition. Prolonged absorption of injectable compositions can be achieved by including in the composition an agent that delays absorption, for example, aluminum monostearate or gelatin.

[0140] The choice of route of administration will depend on whether a local or systemic effect is to be achieved. For example, for local action, the composition can be formulated for topical administration and applied directly to the area where the effect is desired. For systemic, long-term action, the composition can be formulated for enteral administration and administered via the digestive tract. For systemic, immediate and / or short-term action, the composition can be formulated for parenteral administration and administered by a route other than via the digestive tract.

[0141] dosage The compositions of the present disclosure are formulated into acceptable dosage forms by conventional methods known to those skilled in the art. The actual dosage level of the active ingredient in the compositions of the present disclosure may be varied to obtain an amount of the active ingredient effective to achieve the desired therapeutic response for a particular subject, composition, and mode of administration without causing toxicity to the patient. The selected dosage level will depend on various pharmacokinetic factors, including the activity of the particular composition of the present disclosure employed, the route of administration, the time of administration, the rate of release of the particular agent employed, the duration of treatment, other drugs, substances, and / or materials used in combination with the particular composition employed, the age, sex, weight, condition, general health and medical history of the subject being treated, and similar factors well known in the medical field. A physician or veterinarian skilled in the art can easily determine and prescribe the effective amount of the composition required. For example, a physician or veterinarian can begin administering the substance of the present disclosure employed in the composition at a level lower than that required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. Generally, the appropriate daily dose of the composition of the present disclosure will be the amount of substance that is the minimum effective dose to produce a therapeutic effect.Such an effective amount will generally depend on the factors mentioned above.Preferably, the effective daily dose of the therapeutic composition can be administered as 2, 3, 4, 5, 6 or more sub-doses that are administered separately at appropriate intervals throughout the day, optionally in unit dosage form.

[0142] Preferred therapeutic dosage levels are from about 0.1 mg / kg to about 1000 mg / kg (e.g., about 0.2 mg / kg, 0.5 mg / kg, 1.0 mg / kg, 1.5 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 10 mg / kg, 15 mg / kg, 20 mg / kg, 25 mg / kg, 30 mg / kg, 35 mg / kg, 40 mg / kg, 45 mg / kg, 50 mg / kg, 60 mg / kg, 70 mg / kg, 80 mg / kg, 90 mg / kg, 100 mg / kg, 125 mg / kg, 150 mg / kg, 175 mg / kg, 200 mg / kg) of the composition per day administered (e.g., orally or intraperitoneally) to a subject suffering from a disorder described herein (e.g., HBV infection). , 250mg / kg, 300mg / kg, 350mg / kg, 400mg / kg, 450mg / kg, 500mg / kg, 600mg / kg, 700mg / kg, 800mg / kg, 900mg / kg, or 1000mg / kg). Preferred prophylactic dosage levels are from about 0.1 mg / kg to about 1000 mg / kg (e.g., about 0.2 mg / kg, 0.5 mg / kg, 1.0 mg / kg, 1.5 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 10 mg / kg, 15 mg / kg, 20 mg / kg, 25 mg / kg, 30 mg / kg, 35 mg / kg, 40 mg / kg, 45 mg / kg, 50 mg / kg, 60 mg / kg, 70 mg / kg, 80 mg / kg, 90 mg / kg, 100 mg / kg, 125 mg / kg, 150 mg / kg, 175 mg / kg, 200 mg / kg) of the composition per day administered (e.g., orally or intraperitoneally) to a subject. , 250 mg / kg, 300 mg / kg, 350 mg / kg, 400 mg / kg, 450 mg / kg, 500 mg / kg, 600 mg / kg, 700 mg / kg, 800 mg / kg, 900 mg / kg, or 1000 mg / kg). The dose can also be titrated (e.g., the dose can be increased gradually until signs of toxicity such as headache, diarrhea, or nausea appear).

[0143] The frequency of treatment can also vary. The subject can be treated once or more times a day (e.g., once, twice, three times, four times or more) or every many hours (e.g., about every 2, 4, 6, 8, 12, or 24 hours). The composition can be administered once or twice per 24 hours. The time course of treatment can be of varying duration, for example, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more days, 2 weeks, 1 month, 2 months, 4 months, 6 months, 8 months, 10 months, or more than 1 year. For example, treatment can be twice a day for 3 days, twice a day for 7 days, or twice a day for 10 days. The treatment cycle can be repeated at regular intervals, for example, every week, every other month, or every month, and the treatment cycle is separated by a period when no treatment is given. The treatment can be a single treatment, or can continue for the life of the subject (e.g., for many years).

[0144] Patient Selection and Monitoring The methods of the present disclosure described herein require administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof to a subject to activate PRRs for the production of IFNs, ISGs, and cytokines, or to further induce the expression of PRRs (e.g., RIG-I, STING, etc.). In some embodiments, the subject is suffering from or diagnosed with a condition, e.g., a proliferative disease, e.g., cancer. Thus, patients and / or subjects can be selected for treatment with a compound of Formula (I) and / or a pharmaceutically acceptable salt thereof by first evaluating the patient and / or subject to determine whether the subject is infected with a proliferative disease, e.g., cancer. A subject can be evaluated for being infected with a proliferative disease (e.g., cancer) using methods known in the art. For example, the subject can also be monitored after administration of a compound described herein (e.g., a compound of Formula (I) or a pharmaceutically acceptable salt thereof).

[0145] In some embodiments, the subject is a mammal. In some embodiments, the subject is human. In some embodiments, the subject is an adult. In some embodiments, the subject is suffering from a proliferative disease, e.g., cancer. In some embodiments, the subject is suffering from cancer of the breast, bone, brain, cervix, colon, gastrointestinal tract, eye, gallbladder, lymph node, blood, lung, liver, skin, oral cavity, prostate, ovary, penis, pancreas, uterus, testicle, stomach, thymus, thyroid, or other body parts. In some embodiments, the subject is suffering from cancer, including solid tumors (e.g., carcinoma, sarcoma, or lymphoma). In some embodiments, the subject is suffering from hepatocellular carcinoma or other liver cancer. In some embodiments, the subject is suffering from leukemia or other blood cancer. In some embodiments, the subject is suffering from breast cancer, renal cell carcinoma, colon cancer, melanoma, ovarian cancer, head and neck squamous cell carcinoma, pancreatic cancer, prostate cancer, lung cancer, brain cancer, or gastrointestinal stromal cancer. In some embodiments, the subject has cancer cells (eg, tumor cells) that contain a specific cancer-associated antigen that induces a T cell response.

[0146] In some embodiments, the subject is treatment naive. In some embodiments, the subject has previously been treated for a proliferative disease (e.g., cancer). In some embodiments, the subject has relapsed.

[0147] Combination therapy The compounds described herein can be used in combination with other known therapies. "Administered in combination," as used herein, means that two (or more) different therapies are delivered to a subject while the subject is suffering from a disorder, e.g., two or more therapies are delivered after the subject is diagnosed with a disorder, but before the disorder is cured or eliminated, or before the treatments are discontinued for other reasons. In some embodiments, the delivery of one treatment is still occurring when the delivery of the second begins, so there is an overlap in administration. This is sometimes referred to herein as "simultaneous" or "co-delivery." In other embodiments, the delivery of one treatment ends before the delivery of the other treatment begins. In some embodiments, either case, the treatments are more effective because of the combined administration. For example, the second treatment is more effective; e.g., an equivalent effect is seen with the second treatment, or the second treatment reduces symptoms to a greater extent than would be seen if the second treatment were administered without the first treatment, or a similar effect is seen with the first treatment. In some embodiments, delivery is such that the reduction in symptoms or other parameters associated with the disorder is greater than that which would be observed with one treatment delivered in the absence of the other parameter. The effects of the two treatments can be partially additive, wholly additive, or greater than additive. Delivery can be such that the effect of the first treatment delivered is still detectable when the second treatment is delivered.

[0148] The compound described herein and at least one additional therapeutic agent can be administered simultaneously, in the same or separate compositions, or sequentially. For sequential administration, the compound described herein can be administered first and the additional agent can be administered second, or the order of administration can be reversed.

[0149] In some embodiments, the combination of a compound of Formula (I) or a pharmaceutically acceptable salt thereof with an additional agent has a synergistic or additive effect. In some embodiments, the term "additive" refers to a result in which, when two agents are used together, the combination of the agents acts in a manner equal to, but not greater than, the sum of the individual activities of each agent.

[0150] In some embodiments, the combination of a compound of Formula (Ia) or a pharmaceutically acceptable salt thereof with an additional agent has a synergistic or additive effect. In some embodiments, the term "additive" refers to a result in which, when two agents are used together, the combination of agents acts in a manner equal to, but not greater than, the sum of the individual activities of each agent. In some embodiments, the term "synergy" or "synergistic" refers to a result in which, when two agents are used together, the combination of agents requires a lower concentration of each individual agent than would be required to be effective in the absence of the other agent. In some embodiments, a synergistic effect results in a reduction in the minimum inhibitory concentration of one or both agents, such that the effect is greater than the sum of their effects. Synergism is greater than additive. In some embodiments, the agents in the compositions herein may exhibit synergistic activity, where the activity at a particular concentration is at least about 1.25, 1.5, 1.75, 2, 2.5, 3, 4, 5, 10, 12, 15, 20, 25, 50, or 100 times greater than the activity of either agent alone.

[0151] For example, any of the methods described herein may further comprise administering a therapeutically effective amount of an additional agent. Exemplary additional pharmaceutical agents include, but are not limited to, antiproliferative agents, anticancer agents, antidiabetic agents, anti-inflammatory agents, immunosuppressants, and pain-relieving agents. Pharmaceutical agents include drug compounds (e.g., compounds approved by the U.S. Food and Drug Administration as defined in the Code of Federal Regulations (CFR)), peptides, proteins, carbohydrates, monosaccharides, oligosaccharides, polysaccharides, nucleoproteins, mucoproteins, lipoproteins, synthetic polypeptides or proteins, small molecules bound to proteins, glycoproteins, steroids, nucleic acids, DNA, RNA, nucleotides, nucleosides, oligonucleotides, antisense oligonucleotides, lipids, hormones, vitamins, and cells. In some embodiments, the additional agent is an anticancer agent, such as an alkylating agent (e.g., cyclophosphamide).

[0152] In one embodiment, the additional agent is an immuno-oncology agent, e.g., an agent that activates the immune system, e.g., enables it to recognize and destroy cancer cells. Exemplary immuno-oncology compounds are compounds that inhibit immune checkpoint blockade pathways. In one embodiment, the compound is an antibody, such as a PD-1 antibody or a PD-L1 antibody or a costimulatory antibody. In some embodiments, the compound is an anti-CTLA4 antibody. In another embodiment, the agent is a cell-based agent, such as CAR-t therapy. [Example]

[0153] The present disclosure is further illustrated by the following examples and synthetic schemes, which should not be construed as limiting the disclosure in scope or spirit to the specific procedures described herein. It should be understood that the examples are provided to illustrate certain embodiments, and no limitation on the scope of the disclosure is thereby intended. It should further be understood that various other embodiments, modifications, and equivalents thereof that may be apparent to those skilled in the art can be resorted to without departing from the spirit of the present disclosure and / or the scope of the appended claims.

[0154] Abbreviations used in the examples below and elsewhere in this specification are as follows: [Table 3]

[0155] Example 1. Synthesis of exemplary compounds of the present disclosure Procedures for the synthesis of cyclic dinucleotide prodrugs 9 and 4 and cyclic thiodiphosphates [ka] Synthesis of 5'-OH-3'-levulinyl-2'F-dA: Levulinic acid (2.148 g, 18.5 mmol) was dissolved in anhydrous dioxane (50 mL), and the solution was cooled to 5–10 °C in an ice-water bath. DCC (1.939 g, 9.4 mmol) was added in small portions over 1 h. The ice-water bath was removed, and the reaction was allowed to warm to room temperature over 2 h. The resulting dicyclohexylurea precipitate was filtered off and washed with anhydrous dioxane (10 mL). The filtrate was added to a solution of 5'DMT-2'F-3'OH-dA (5.0 g, 7.4 mmol) in anhydrous pyridine (50 mL), followed by the addition of a catalytic amount of DMAP under an argon atmosphere. After stirring at room temperature for 2 h, the mixture was evaporated to dryness. The residue was dissolved in DCM (150 mL) and the organic phase was washed with 5% NaHCO.sub.3 (100 mL) and brine (100 mL), dried over Na.sub.2SO.sub.4 and concentrated under reduced pressure to give the desired product as a white solid. [ka]

[0156] Detritylation: The above solid was dissolved in DCM (100 mL), and water (1.33 mL, 74 mmol) was added to the reaction mixture. 6% DCA in DCM (100 mL) was then added, and the reaction mixture was stirred at room temperature for 10–15 min. The resulting mixture was quenched by the addition of methanol (25 mL) and then washed with 5% NaHCO3 solution (150 mL) and brine (150 mL). The combined organic layers were dried over Na2SO4 and concentrated under reduced pressure. The crude residue was purified using CombiFlash silica gel column chromatography eluting with 0–5% MeOH in DCM to afford 3.45 g (62% yield) of the pure desired product as a white solid. [ka]

[0157] Coupling: A mixture of 5'OH-3'-levulinylated-2'F-deoxy-azinosine (700 mg, 1.48 mmol) and 5'DMT-2'F-3'CED-phosphoramidite-deoxy-uridine (1.66 g, 2.22 mmol) was dried under high vacuum for 1-2 hours. The round-bottom flask containing the reaction mixture was flushed with argon. Anhydrous acetonitrile (40 mL) was added to the reaction mixture, followed by ETT (279 mg, 2.146 mmol) in acetonitrile (5.0 mL) under an argon atmosphere. The resulting mixture was stirred at room temperature under argon for 2 hours. When TLC analysis indicated the reaction was complete, water (80 μL, 2 equivalents relative to the amidite) was added.

[0158] Sulfurization: In a silanization flask, Beaucage reagent (3H-BD) (592 mg, 2.96 mmol) was dissolved in acetonitrile (5.0 mL). The previous coupling reaction mixture was transferred to a solution of sulfurization reagent (3H-BD) in acetonitrile under an argon atmosphere. The resulting mixture was stirred at room temperature for 45 minutes to complete the sulfurization reaction. After adding methanol (10 mL), the reaction mixture was stirred for 30 minutes. The resulting mixture was evaporated to dryness under reduced pressure. The crude residue was dissolved in DCM (100 mL) and washed with water (75 mL). The DCM layer was separated, dried over Na2SO4, and used in the detritylation step.

[0159] Detritylation: The previously obtained DCM layer containing the sulfurized product was cooled in an ice-water bath. A 5% PTSA solution in DCM:MeOH (7:3, 100 mL) was added, and the reaction mixture was stirred for 15 minutes to complete the detritylation reaction. Water (50 mL) was then added, and the resulting mixture was stirred for another 15 minutes. The reaction mixture was transferred to a separatory funnel, and the layers were separated. The organic layer was washed with 5% NaHCO3 solution (100 mL), while the pH of the aqueous layer was above 7.0. The combined organic layers were dried over Na2SO4 and concentrated under reduced pressure to obtain the crude product. The crude product was purified using CombiFlash silica gel column chromatography, eluting with 0–5% MeOH in DCM, to obtain 960 mg of pure desired product as a white solid. [ka]

[0160] Levulinyl group deprotection: The 3'-levulinyl-protected dinucleotide thiophosphate was treated with 0.5 M hydrazine monohydrate in a mixture of pyridine:acetic acid (3:2), and the reaction mixture was stirred at room temperature for 15 min. When TLC analysis showed the reaction was complete, 2,4-pentanedione (2.0 mL) was then added to quench unreacted hydrazine hydrate. The volatiles were removed under reduced pressure, and the reaction mixture was partitioned between 25% IPA in DCM (50 mL) and water (50 mL). The organic layer was collected and evaporated to dryness under reduced pressure to give a thick liquid, which was co-evaporated with toluene (2 × 15 mL) to give a crude residue, which was purified by Combi-Flash silica gel column chromatography using 0–10% MeOH in DCM to give 725 mg of pure desired product as a white solid.

[0161] Cyclization: Dinucleotide phosphorothioate trimer (1 equivalent) and 2-cyanoethyl tetraisopropyl phosphorodiamidite (bisamidite) (1 equivalent) were dissolved in a mixture of anhydrous acetonitrile and anhydrous DCM (2:1, 30 mL). Under an inert atmosphere, diisopropylaminotetrazolide (1 equivalent) was added to the reaction mixture in four portions over 1 hour. The solution was stirred at room temperature for an additional 2 hours, and then ETT (2.0 equivalents) was added to the reaction mixture and stirred overnight. Deoxygenated water (29 μL) was then added to the reaction mixture. [ka]

[0162] Sulfurization (Synthesis of Protected Cyclic Phosphorothiodiphosphate): Beaucage reagent (3H-BD) (2.0 equiv.) was dissolved in acetonitrile in a silanization flask. A portion (two-thirds) of the previous cyclization product was added to the sulfurization reagent under an argon atmosphere. The reaction mixture was stirred at room temperature for 45 min. Methanol (10 mL) was then added, and the resulting mixture was stirred for 30 min. The solvent was evaporated under reduced pressure, and the crude residue was dissolved in DCM (50 mL) and washed with water (50 mL). The DCM layer was separated, dried over Na2SO4, and concentrated under reduced pressure. The crude product was purified using Combiflash silica gel column chromatography eluting with 0-10% MeOH in DCM to give 150 mg of pure desired product. [ka]

[0163] Oxidation (synthesis of protected cyclic phosphoromonothiodiphosphate): TBHP (4.0 equiv.) was added to a stirred solution of the second portion (one-third) of the cyclized product at 0 °C, and the reaction mixture was allowed to warm to room temperature over 15 min. Excess TBHP was quenched by the addition of saturated sodium bisulfite solution, and the resulting mixture was evaporated under reduced pressure. The crude residue was dissolved in DCM (25 mL) and washed with water (20 mL). The organic layer was separated, dried over Na2SO4, and concentrated under reduced pressure. The resulting crude product was purified using Combiflash silica gel column chromatography eluting with 0-10% MeOH in DCM to give 60 mg of pure desired product. [ka]

[0164] Deprotection of cyclic phosphorothiodiphosphate [Synthesis of compound 2]: The protected cyclic phosphorothiodiphosphate (60 mg) was dissolved in concentrated NHOH (2.0 mL) and stirred at room temperature overnight. When LCMS showed the reaction was complete, the mixture was evaporated under reduced pressure to remove ammonia. The aqueous layer was washed with ethyl acetate (5 × 5 mL), separated, and lyophilized to give 100 mg of crude product as a white fluffy solid. [ka]

[0165] Alkylation of cyclic phosphorothiodiphosphate [Synthesis of compound 4]: Cyclic phosphorothiodiphosphate (25 mg) was dissolved in water (250 μL). Then, a solution of 4-(iodomethyl)phenyl 4-(decyloxy)benzoate (42 mg) in a mixture of THF:acetone (1:1, 2.0 mL) was added. The pH of the reaction mixture was approximately 3.5-4.0. The reaction mixture was stirred at room temperature for 40 h. The crude product was purified using Combiflash silica gel column chromatography eluting with 0-10% IPA in DCM to give 25 mg of the desired product as a tan solid. [ka]

[0166] Deprotection of cyclic phosphoromonothiodiphosphate [Synthesis of compound 3]: The protected cyclic phosphoromonothiodiphosphate (60 mg) was dissolved in concentrated NHOH (5.0 mL) and then stirred at room temperature overnight. When LCMS indicated a reaction, the mixture was evaporated under reduced pressure to remove ammonia. The aqueous layer was washed with ethyl acetate (5 × 5 mL), separated, and lyophilized to give 50 mg of the desired crude product as a white, fluffy solid. [ka]

[0167] Alkylation of Cyclic Phosphoromonothiodiphosphate [Synthesis of Compound 1]: Cyclic phosphoromonothiodiphosphate (20 mg) was dissolved in water (200 μL). A solution of 4-(iodomethyl)phenyl 4-(decyloxy)benzoate (18 mg) in a mixture of THF:acetone (1:1, 1.4 mL) was then added. The pH of the reaction mixture was approximately 4.0. The reaction mixture was stirred overnight at room temperature, and the solvent was removed under reduced pressure. The resulting crude residue was redissolved in water:acetonitrile (1:1, 2.0 mL). A precipitate (unreacted alkylating reagent) formed and was removed by centrifugation. The mother liquor was lyophilized, and the crude product was purified by CHCl3 containing 0.2 M ammonium acetate buffer. 18 Purification was performed using a Sep-Pak column (Waters, 4.0 g). The compound was eluted with acetonitrile:water (1:1). Pure fractions were collected and lyophilized to give 5-6 mg of the pure desired product as a fluffy white solid.

[0168] Example 2. In vitro activation of ISG54 and NF-κβ in HEK293 cells In this experiment, to screen for potential STING agonists, HEK293 cells (SZ14) stably expressing either the ISG54 ISRE-luc reporter or the NF-κβ-luc reporter gene were treated in duplicate with exemplary compounds of the present disclosure or 2',3'-cGAMP as a control in digitonin buffer for 5 hours. ISG54 or NF-κβ activity was measured using the Steady-glo buffer system (Promega) and the EC values ​​are summarized in Table 3 below. 50 Generally, the half-maximal effective concentration (EC 50 ) refers to the concentration of drug that induces a response halfway between baseline and maximum after a specified exposure time. This calculation is applicable to compounds with enzyme inhibitory activity, since the baseline for untreated samples can be set at 100% enzyme activity, and therefore, the percent inhibition is evaluated based on this 100% maximum standard. For these tests, EC 50 Values ​​relate to the concentration required to achieve a value of 50% activity level above the untreated sample, which was set at 0%.

[0169] In Table 3, "A" indicates an EC<50 nM 50 "B" represents an EC between 50 nM and 500 nM 50 and "C" is an EC between 500 nM and 1 μM 50 and "D" is an EC between 1 μM and 2 μM 50 and "E" is an EC50 value greater than 2 μM 50 Data are presented as fold induction over cells receiving DMSO (compound carrier) alone, as the mean + / - standard deviation of replicate wells per stimulant. [Table 4] TIFF0007749629000054.tif50163

[0170] Example 3. Evaluation of IRF-type I IFN activity in THP cells THP1-double cells were treated in triplicate with an exemplary compound of the present disclosure (e.g., Compound 2 or Compound 3) in lipofectamine or 2',3'-cGAMP in lipofectamine as a control at various concentrations for 22 hours. The levels of IRF-inducible luciferase reporter activity in cell culture supernatants were assayed using Quanti-luc reagent and are summarized in Figure 9. Data are presented as the mean + / - standard deviation of duplicate wells per stimulant and as fold induction relative to cells receiving DMSO (compound carrier) alone.

[0171] Example 4. Measurement of cytotoxicity of exemplified compounds The cytotoxicity of exemplary compounds in THP1 cells was assessed using the CellTiter-Glo assay (Promega). THP1 bilayer cells grown in complete medium were treated with various concentrations of compounds or DMSO control. CellTiter-Glo® luminescent cell viability / cytotoxicity was measured by assessing the number of viable cells in culture based on quantification of ATP present via a "glow-type" luminescent signal generated by the luciferase reaction. Percent apoptosis was calculated from the fold change in luminescence compared to DMSO-treated samples.

[0172] Example 5. Quantification of STING binding SZ14 HEK293 cells stably expressing the ISG54 ISRE-luc reporter gene were treated with the exemplary compound 1,2'3'-cGAMP (a natural STING ligand) or DMSO for 5-6 hours in the presence of digitonin. ISRE-luciferase activity was measured and normalized to DMSO-treated cells (mean ± standard deviation of triplicate wells per stimulant).

[0173] Alternatively, live ISG-Dual cells in 96-well plates were stimulated in triplicate with compound / lipoprotein, cGAMP / lipoprotein complexes, or compound alone at 37°C and 5% CO for 22–24 h. Secreted luciferase activity in cell culture supernatants was measured using Invivogen Quanti-luc. Data are presented as fold induction relative to DMSO-treated cells (mean ± standard deviation of triplicate wells per stimulus).

[0174] Example 6. Induction of Type III IFN (IL-29) Production in THP Cells by Exemplary Compounds THP1 double (wild-type) cells were treated in triplicate with the exemplary compounds alone or with cGAMP / Lipo for 21 hours. IL-29 levels in the culture supernatants were measured using ELISA. Results shown are the mean ± standard deviation of duplicate wells.

[0175] Example 7. Figure 9 shows that Compound 1 induces cell death by apoptosis. Apoptosis of THP1 cells was assessed using the Caspase-Glo® 3 / 7 assay (Promega). THP1 bilayer cells grown in complete medium were treated with various concentrations of Compound 1 or 2'3-cGAMP containing Lipofectamine LTX or DMSO control. Caspase-3 and caspase-7 activity was measured using a proluminescent caspase-3 / 7 substrate containing the tetrapeptide sequence DEVD, which was cleaved to release amino-luciferin, the luciferase substrate used for light generation. After 20 hours of incubation, apoptotic activity was assessed by measuring the level of amino-luciferin. The percent apoptosis was calculated from the fold change in luminescence compared to DMSO-treated samples. CC50 values ​​were generated by curve fitting with Xlfit.

[0176] Example 8. Figure 10 shows the selective induction of apoptosis by Compound 1 in an acute monocytic leukemia cell line (THP1) versus PBMCs. Apoptosis in THP1 cells and PBMCs was assessed using the Caspase-Glo® 3 / 7 assay (Promega). THP1 cells and PBMCs grown in complete medium were treated with various concentrations of Compound 1 or 2'3-cGAMP containing Lipofectamine LTX or DMSO control. Caspase-3 and caspase-7 activity was measured using a proluminescent caspase-3 / 7 substrate containing the tetrapeptide sequence DEVD, which was cleaved to release amino-luciferin, the luciferase substrate used for light generation. After 20 hours of incubation, apoptotic activity was assessed by measuring the level of amino-luciferin. The percentage of apoptosis was calculated from the fold change in luminescence compared to DMSO-treated samples.

[0177] Example 9. Figure 11 shows that Compound 1 induces selective and enhanced induction of ISG and PRR-related genes in acute monocytic leukemia cell line (THP1) compared to primary PBMC. Gene expression analysis in THP1 and PBMC: THP1 cells and PBMC grown in complete medium were treated with either 5 μM Compound 1 or 2'3-cGAMP containing Lipofectamine LTX or DMSO control. After 20 hours of incubation, RNA was extracted, and gene expression of different interferon-stimulated genes (ISGs) and various pattern recognition receptors (PRRs) was evaluated by real-time PCR. Fold induction was calculated using the ΔΔct method.

[0178] Example 10. Figure 12 shows that Compound 1 inhibits tumor cell growth. Tumor cells in 96-well plates were treated with Compound 1 (without lipofectamine) or recombinant IFN (U-IFN) once a day for 3 days. Cells were fixed with 1% paraformaldehyde and stained with DAPI. Cells were automatically imaged using ImageXpress, and the total number of viable cells was analyzed using MetaXpress software. Results are shown as the total number of cells per group or as a percentage reduction calculated by normalizing to DMSO-treated cells.

[0179] Example 11. Figure 21 shows that compound 4 has enhanced activity in acute monocytic leukemia cell line (THP1) compared to primary cell PBMC. Gene expression analysis in THP1 and PBMC: THP1 cells and PBMC grown in complete medium were treated with either 5 μM compound 4 or 2',3'-cGAMP containing Lipofectamine LTX or DMSO control. After 20 hours of incubation, RNA was extracted, and gene expression of different interferon-stimulated genes (ISGs) and various pattern recognition receptors (PRRs) was evaluated by real-time PCR. Fold induction was calculated using the ΔΔct method.

[0180] Example 12. Efficacy of exemplary compounds via intraperitoneal administration in a breast cancer model. The efficacy of intraperitoneal administration of Compound 1 was investigated in the 4T1.luc2 orthotopic mouse breast cancer model. Thirty female BALB / c mice, 7-10 weeks old, were randomly divided into four treatment groups based on their body weight after one day and treated according to the dosing schedule outlined in Table 4 below. Compound 1 was dissolved in saline and administered at 10 mL / kg (0.200 mL / 20 g animal), with a cell injection volume of 0.05 mL / animal. [Table 5]

[0181] Each animal was monitored individually. The experimental endpoint was a tumor volume of 2000 mm or 45 days. Groups 1 and 2 animals underwent whole-body bioluminescence imaging starting on day 5 and then weekly (days 12, 19, 26, 33, and 41). At the endpoint, blood and tissues (lungs, lymph nodes, spleen, and tumor) were analyzed for the presence of metastases and biomarker (CD45, CD3, CD4, CD8, CD11b, CD25, Ly-6G, Ly-6C, FoxP3) levels. As shown in Figure 70, mice treated with Compound 1 showed significantly reduced tumor growth compared to controls.

[0182] Example 13. Determination of the maximum tolerated dose of an exemplary compound administered orally. To determine the maximum tolerated dose of orally administered compound, 15 female BALB / c mice aged 7-10 weeks were divided into three treatment groups. Each group received either Compound 1 or vehicle orally according to the schedule outlined in Table 5 below. Compound 1 was administered at 10 mL / kg (0.200 mL / 20 g of animal). Oral administration of Compound 1 up to 60 mg / kg / day once or twice daily resulted in no adverse clinical signs, and the compound was well tolerated, as shown in Table 5. [Table 6]

[0183] equivalent The disclosures of each and every patent, patent application, and publication cited herein are incorporated herein by reference in their entirety. While the present disclosure has been described with respect to certain embodiments, it will be apparent that other embodiments and variations may be devised by those skilled in the art without departing from the true spirit and scope of the present disclosure. It is intended that the appended claims be construed to include all such embodiments and equivalent variations. Any patent, publication, or other disclosure material said to be incorporated herein by reference is incorporated herein in whole or in part only to the extent that it does not contradict existing definitions, statements, or other disclosure material set forth in this disclosure. As such, to the extent necessary, the present disclosure as expressly set forth herein takes precedence over any conflicting material incorporated herein by reference.

[0184] While the present disclosure has been particularly shown and described with respect to preferred embodiments thereof, it will be understood by those skilled in the art that various changes in form and details can be made therein without departing from the scope of the present disclosure as encompassed by the appended claims.

Claims

1. Use of a compound in the manufacture of a medicament for i) treating cancer, ii) immunomodulating, or iii) inducing an immune response, wherein the compound is represented by formula (I): 【Chemical 1】 or a pharmaceutically acceptable salt or stereoisomer thereof, wherein: Z is either S or O; B １ teeth, 【Chemistry 2】 and B ２ teeth, 【Chemistry 3】 and X １ and X ２ each is independently O or S; Y １ and Y ２ One of the groups is O, S, or NR ５ and Y １ and Y ２ The other is S or NR ５ and L １ and L ２ Each of is independently absent or C １ ~C ６ alkyl or C １ ~C ６ heteroalkyl, wherein alkyl and heteroalkyl are each optionally selected from R ６ is replaced by R １ and R ２ each independently represents hydrogen, halo, —CN, C １ ~C ２０ Alkyl (e.g., C １ ~C ６ alkyl), or OR ７ and R ３ and R ４ Each of the groups is independently hydrogen, C １ ~C ２０ Alkyl (e.g., C １ ~C ６ alkyl), C １ ~C ２０ Heteroalkyl (e.g., C １ ~C ６ heteroalkyl), OC(O)OC １ ~C ２０ Alkyl (e.g., C １ ~C ６ alkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each optionally have one or more R ８ is replaced by R ５ is hydrogen or C １ ~C ２０ Alkyl (e.g., C １ ~C ６ alkyl), R ６ But halo, -CN, C １ ~C ２０ Alkyl (e.g., C １ ~C ６ alkyl), OR ７ , oxo, cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein the alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl each may be joined by one or more R ９ and optionally replaced by R ７ is hydrogen, C １ ~C ２０ Alkyl (e.g., C １ ~C ６ alkyl), cycloalkyl, heterocyclyl, aryl, or heteroaryl, each of which may optionally be joined by one or more R ９ is replaced by Each R ８ are independently １ ~C ２０ Alkyl (e.g., C １ ~C ６ alkyl), C １ ~C ２０ Heteroalkyl, C(O)—C １ ~C ２０ Alkyl, OC(O)-C １ ~C ２０ Alkyl (e.g., C １ ~C ６ alkyl), C(O)O-C １ ~C ２０ Alkyl (e.g., C １ ~C ６ alkyl), OC(O)O-C １ ~C ２０ Alkyl (e.g., C １ ~C ６ alkyl), C(O)N(R ５ )-C １ ~C ２０ Alkyl (e.g., C １ ~C ６ alkyl), N(R ５ ) C(O)-C １ ~C ２０ Alkyl (e.g., C １ ~C ６ alkyl), OC(O)N(R ５ )-C １ ~C ２０ Alkyl (e.g., C １ ~C ６ alkyl), O-aryl, O-heteroaryl, C(O)-aryl, C(O)-heteroaryl, OC(O)-aryl, C(O)O-aryl, OC(O)-heteroaryl, C(O)O-heteroaryl, C(O)O-aryl, C(O)O-heteroaryl, C(O)N(R ５ )-aryl, C(O)N(R ５ )-heteroaryl, N(R ５ )C(O)-aryl, N(R ５ ) ２ C(O)-aryl, or N(R ５ ) C(O)-heteroaryl, S(O) ２ N (R ５ )-aryl, wherein alkyl, heteroalkyl, aryl, and heteroaryl each have one or more R ９ and optionally replaced by And each R ９ are independently １ ~C ２０ Alkyl, O-C １ ~C ２０ Alkyl, C １ ~C ２０ heteroalkyl, halo, —CN, OH, oxo, aryl, heteroaryl, O-aryl, or O-heteroaryl.

2. The compound is a compound of formula (I-b), formula (I-c), formula (I-d), or formula (I-e), 【Chemistry 4】 or a pharmaceutically acceptable salt thereof, １ , B ２ , X １ , X ２ , Y １ , Y ２ , L １ , L ２ , R １ , R ２ , R ３ , R ４ and each of these subvariables is defined in claim 1.

3. R １ and R ２ each independently represents hydrogen, halo, or OR ７ 3. The use according to claim 1 or 2, wherein

4. R １ and R ２ The use of any one of claims 1 to 3, wherein each of is independently halo (e.g., fluoro).

5. X １ and X ２ The use according to any one of claims 1 to 4, wherein each of is independently O.

6. Y １ and Y ２ One of the groups is O, and Y １ and Y ２ The use according to any one of claims 1 to 5, wherein the other is S.

7. Y １ and Y ２ The use according to any one of claims 1 to 5, wherein each of

8. Y １ is S and Y ２ The use according to any one of claims 1 to 5, wherein is O.

9. L １ and L ２ Each of the is independently C １ ~C ６ Alkyl (e.g., CH ２ 9. The use according to any one of claims 1 to 8, wherein

10. R ３ and R ４ is independently hydrogen, aryl, or heteroaryl, and the aryl and heteroaryl are selected from 1 to 5 R ８ The use according to any one of claims 1 to 9, optionally substituted by

11. R ３ are aryl or heteroaryl, each of which is 1 to 5 R ８ and optionally substituted by R ４ The use according to any one of claims 1 to 9, wherein is hydrogen.

12. R ３ There is one R ８ phenyl substituted with R ４ The use according to any one of claims 1 to 9, wherein is hydrogen.

13. R ３ and R ４ Each of the groups independently represents one R ８ The use according to any one of claims 1 to 9, wherein the phenyl is phenyl substituted with

14. R ８ But 1 to 5 R ９ (For example, one R ９ 14. The use according to any one of claims 1 to 13, wherein the aryl is OC(O)-aryl optionally substituted by

15. R ９ But O-C １ ~C １２ Alkyl (e.g., O—CH ２ (CH ２ ) ８ CH ３ The use according to any one of claims 1 to 14, wherein

16. R ３ and R ４ The use according to any one of claims 1 to 9, wherein each of is hydrogen.

17. Y ２ is O and L ２ does not exist, and R ４ The use according to any one of claims 1 to 5, wherein is hydrogen.

18. Y １ is S and Y ２ is O and L １ and L ２ each of which is independently absent, and R ３ and R ４ The use according to any one of claims 1 to 5, wherein each of is independently hydrogen.

19. Use of a compound in the manufacture of a medicament for i) treating cancer, ii) immunomodulating, or iii) inducing an immune response, wherein the compound is: 【Chemistry 5】 or a pharmaceutically acceptable salt thereof.

20. The use according to any one of claims 1 to 19, wherein the medicament is for the treatment of cancer.

21. 21. The use of claim 20, wherein the cancer is cancer of the breast, bone, brain, cervix, colon, digestive tract, eye, gallbladder, lymph node, blood, lung, liver, skin, oral cavity, prostate, ovary, penis, pancreas, uterus, testicle, stomach, thymus, thyroid, or other part of the body.

22. 21. The use according to claim 20, wherein the cancer is cancer of the liver.

23. The use of claim 20, wherein the medicament is administered in combination with an additional active agent (e.g., an anticancer agent).

24. 24. The use of claim 23, wherein the additional agent comprises methotrexate, 5-fluorouracil, doxorubicin, vincristine, bleomycin, vinblastine, dacarbazine, toposide, cisplatin, epirubicin, or sorafenib tosylate.

25. The use of any one of claims 1 to 19, wherein the medicament is for immunomodulation.

26. The use of any one of claims 1 to 19, wherein the medicament is for inducing an immune response.

27. 27. The use of claim 26, wherein the immune response comprises anti-tumor immunity.

28. 27. The use of claim 26, wherein the immune response comprises induction of a PRR (e.g., STING, RIG-I, or MDA5).

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