Pyridinyl-substituted oxoisoindoline compounds

Abstract

Formula (I): TIFF2024515243000053.tif39153[where, R 1 , R 2 , R 4 , R 6 , m and n are defined in this disclosure. or salts thereof. Also disclosed are methods of using the compounds, and pharmaceutical compositions containing the compounds, to inhibit Helios protein. These compounds are useful in the treatment of viral infections and proliferative diseases (e.g., cancer).

Description

[Technical field]

[0001] (Related Applications) This application claims priority to U.S. Provisional Application No. 202111016193, filed April 6, 2021, and U.S. Provisional Application No. 202111022098, filed May 17, 2021, all of which are incorporated by reference herein.

[0002] (explanation) The present invention generally relates to pyridinyl-substituted oxoisoindoline compounds that inhibit the Helios protein. The present invention provides pyridinyl-substituted oxoisoindoline compounds, compositions containing the compounds, and methods of using them. The present invention further relates to pharmaceutical compositions containing at least one compound according to the present invention, which are useful for treating proliferative diseases (e.g., cancer) and viral infections. [Background technology]

[0003] Regulatory T cells (Tregs) play an essential role in maintaining self-tolerance and immune homeostasis by suppressing and maintaining immune unresponsiveness against intense immune and inflammatory responses. By maintaining a phenotype that exhibits a stable suppressive effect of immune unresponsiveness, Tregs suppress excessive immune responses and prevent or alleviate autoimmune diseases. Numerous reports have demonstrated the presence of Tregs in human tumor tissues. Studies have shown a clear negative correlation between the number of Tregs and the infiltration and survival rate of T cells in tumors (Curiel et al., 2004, Nat. Med. 10:942-949;Viguier et al., 2004, J Immuno. 1173:1444-1453;Beyer et al., 2006, Blood 108:804-811;Zou et al., 2006, Nat. Rev. Immunol. 6:295-307), implying that Tregs potentially play an important role in inhibiting effective antitumor immunity. Previous evidence has shown that Foxp3+CD25+CD4+Tregs predominantly infiltrate tumors and apparently inhibit immune responses against tumor cells in rodents and humans. Once activated by a specific antigen, Tregs behave in an antigen-nonspecific manner in vitro, suppressing killer T cells by overlooking them (Takahashi et al., 1998, Int Immunol. 10:1969-80;Thornton et al., 1998, J Exp. Med. 188:287-96). Foxp3+CD25+CD4+Tregs can apparently suppress a broad range of antitumor immune responses, including CD4+helper T cells, CD8+T cells, natural killer cells, and natural killer T cells (Tanaka et al., 2017, Cell Research 27:109-118). Depletion of CD25+CD4+Tregs in tumors alters the cytokine environment in tumor sites, resulting in regression of established tumors (Yu et al., 2005, J Exp Med. 201:779-91).Moreover, transplantation of CD4+ T cells deficient in Tregs significantly increases antitumor immune responses compared with transplantation of Tregs containing T cells (Antony et al., 2005, J Immunol 174:2591-601). Tumor-infiltrated Tregs activated by either tumor-derived self-antigens or tumor-associated antigens can similarly suppress specific antitumor immune responses. This means that modulating the activity of key factors to control Treg differentiation is a promising strategy for treating certain diseases, such as cancer and viral infections.

[0004] FoxP3+CD4 Tregs are highly stable. Research is still ongoing to understand this genetic mechanism to see if the phenotype is stable even after the spread of inflammation, infection, or autoimmune disease. Transcription factors (TFs) involved in maintaining the stable immunosuppressive phenotype of Tregs can be used in this study. The Helios (IKZF2) gene, a member of the Ikaros family of TFs, differs from other Ikaros family members on the basis of its selective expression by negatively selected thymocytes, as well as CD4 and CD8 T cells of the suppressive lineage. Helios is expressed by two regulatory T cell lineages, FoxP3+CD4+ and Ly49+CD8+ Tregs, which are essential lineages for maintaining self-tolerance (Kim et al., 2015, Science 350:334-339;Sebastian et al., 2016, J Immunol 196:144-155). Interestingly, recent studies have suggested that Helios is dispensable for Treg activity in the steady state, but is essential for regulating the genetic program of FoxP3+CD4 Tregs during inflammation to maintain a stable phenotype and enhance suppressive function (Thornton et al., 2010, J Immunol. 184:3433-3441; Kim et al., 2015). Expression of Helios was shown to be crucial for Tregs to maintain an immunosuppressive and immunounresponsive phenotype against strong inflammatory responses. Activating the IL-2Rα-STAT5 pathway was shown to be a key associated factor for Treg survival and stability (Kim et al., 2015). Helios exerts a major lymphocyte-intrinsic inhibitory activity, and thus plays an essential role in maintaining the phenotype of FoxP3+CD4 Tregs to prevent autoimmune diseases in the presence of highly activated autoreactive T cells from Scurfy mice lacking the forkhead domain of FoxP3. Bone marrow chimeras reconstituted with Helios- / - / Scurfy bone marrow (BM) (non-Helios+) / + / Scurfy BM cells rapidly established autoimmunity ( Kim et al., 2015 ).These results indicate that Helios is important for the selection, differentiation, and functional expression of autoreactive T cells. Immunosuppression by Tregs can impede antitumor immune responses. Selective deficiency of Helios in FoxP3+CD4 Tregs leads to Treg instability and the transformation of intratumoral CD4 Tregs into effector T cells (Teff). Intratumoral Treg instability can lead to Treg conversion and reduced suppressive activity of Tregs, resulting in increased numbers of Teff cells in the tumor. Furthermore, poor IL-2 response was observed in intratumoral Tregs lacking Helios. This may result in a reduced number of activated Tregs and also contribute to increased intratumoral Teff activity. Interactions between tumor cells and infiltrating immune cells promote the secretion of inflammatory mediators (such as TNF-α, IL-6, IL-17, IL-1, and TGF-β) to form a local inflammatory environment (Kim et al., 2015).

[0005] Lineage instability of Helios-deficient Tregs is also caused by reduced FoxP3 expression, resulting in the production of inflammatory cytokines and effector T cells. The transformation of Helios-deficient Tregs into effector cells within the tumor tissue microenvironment is associated with increased expression of genes governing the Teff phenotype (Yates et al., 2018, PNAS, 2018, 115:2162-2167). Helios deficiency results in an unstable phenotype only within the tumor microenvironment (TME), but not in peripheral lymphoid organs (Nakagawa et al., 2016, PNAS 113:6248-6253). In chronically inflamed TME, Helios deficiency in Tregs increases helper T cells for TFs and effector cytokines, and can significantly alleviate the suppressive gene program for helper T cell differentiation. A subpopulation of Tregs with high affinity for self-antigens showed increased GITR / PD-1 expression and increased responsiveness to self-antigens, which are evident in the genetic alterations of these Helios-deficient Tregs, which combined promote the conversion of Tregs to Teffs in the TME and increase T cell receptor (TCR) engagement and co-stimulatory receptor expression by Tregs, suggesting that the gene expression alterations that are a key feature of Treg conversion vary depending on the immune environment (Yates et al., 2018).

[0006] Decreased Helios expression in FoxP3+CD4 Tregs may convert memory Tregs into Teff cells expressing tumor antigen-specific autoreactive T cell receptors. In chronic inflammatory conditions due to tumor growth, the converted Treg characteristics may be selectively induced. Helios-deficient Tregs express TCRs with high affinity for self-peptides / MHC, which may promote stable activation of Tregs in the TME (Yates et al., 2018). Given the increased autoreactivity of TCRs in CD4 Tregs compared with conventional T cells, the conversion of Tregs may produce highly potent effector CD4 T cells, along with suppressing the suppressive effect mediated by Tregs in the TME. Moreover, a method to selectively convert intratumoral Tregs into Teff cells without affecting the systemic Treg population would be an even more effective strategy. As a key factor for maintaining Treg abundance and functional stability in response to various immune responses, Helios may be pharmacologically relevant to strategies to enhance current tumor immunotherapy. Because the change from Tregs to Teffs can be restricted to the inflammatory tumor microenvironment, antibody or small molecule-based drugs targeting Helios may lead to improved immunotherapy of Treg-dependent cancers. Importantly, the change in Helios-deficient Tregs occurs only within the local inflammatory environment of the tumor. This approach may avoid autoimmune side effects associated with the reduction of systemic Tregs. Therefore, a strategy to specifically suppress Helios-dependent regulation of intratumoral Tregs would be a critical support for improving cancer immunotherapy. Furthermore, it has been reported that depletion of Foxp3+Tregs enhances vaccine-induced antitumor T cell responses (Nishikawa et al., 2010, Int. J. Cancer 127:759-767). This suggests that reducing Helios levels could be effective in enhancing the efficacy of cancer vaccines.

[0007] Antitumor immunotherapy during viral infections may limit the immune response of Treg cells driven by excessive inflammation and inhibit effective antiviral T cell responses, promoting viral persistence (Schmitz et al., 2013, PLOS Pathogens 9:e1003362). Chronic infection of mice with lymphocytic choriomeningitis virus resulted in a significant expansion of Foxp3+ Tregs, implying a potential mechanism by which certain infectious agents may evade the host immune response by activating and expanding Tregs (Punkosdy et al., 2011, PNAS 108:3677-3682). During chronic virus-related diseases, reducing the levels of Helios in activated Tregs may have therapeutic benefits.

[0008] There is a need for compounds that are useful as inhibitors of Helios proteins.

[0009] SUMMARY OF THE PRESENTINVENTION The present invention provides pyridinyl-substituted oxoisoindoline compounds of formula (I) or salts thereof useful for reducing Helios protein levels and Helios activity levels and / or inhibiting Helios expression levels in a cell.

[0010] The present invention also provides a pharmaceutical composition comprising a compound of formula (I) and / or a pharma- ceutically acceptable salt thereof; and a pharma- ceutically acceptable carrier.

[0011] The present invention also provides a method for treating a disease or disorder caused by a decrease in the activity of the Helios protein, the method comprising administering to a patient a compound of formula (I) and / or a pharma- ceutical acceptable salt thereof.

[0012] The present invention also provides processes and intermediates for use in the preparation of compounds of formula (I) and / or salts thereof.

[0013] The present invention also provides a compound of formula (I) and / or a pharma- ceutically acceptable salt thereof for use in therapy.

[0014] The present invention also provides the use of a compound of formula (I) and / or a pharma- ceutical acceptable salt thereof for the manufacture of a medicament for the treatment of certain diseases (such as cancer and viral infections) to reduce Helios protein levels and Helios activity levels in cells and / or suppress Helios expression levels to control Treg differentiation.

[0015] The compounds of formula (I) and compositions containing the compounds of formula (I) may be used to treat, prevent or cure viral infections and various proliferative diseases, such as cancer. Pharmaceutical compositions containing these compounds are useful for treating, preventing or inhibiting the progression of diseases or disorders, such as viral infections and cancer, in various treatment sites.

[0016] These and other features of the present invention are set forth in the broader disclosure. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0017] Applicant has discovered substituted oxoisoindoline compounds that inhibit Helios protein by promoting the interaction of Helios protein and its corresponding E3 ubiquitin ligase complex (Cullin4-Cereblon, CUL4-CRBN). These compounds reduce Helios protein levels and Helios activity levels in cells and / or suppress Helios expression levels to control Treg differentiation. These compounds are useful for treating certain diseases, such as cancer and viral infections. These compounds provide useful pharmaceuticals with desirable stability, bioavailability, therapeutic index, and toxicity values ​​important to their druggability.

[0018] The first aspect of the present invention relates to a compound having at least one formula (I): [ka] [In the formula, R1 is -NH2 or -NH(CH3); Each R2 is independently F, Cl, -CN, C 1-4 alkyl, -CH2F, -CHF2, -CF3, -OCH3 or cyclopropyl; Each R4 is independently F, Cl, -CH3, -CH2F, -CHF2, -CF3, or -OCH3; R6 is hydrogen, C 1-2 Alkyl or C 1-2 is a fluoroalkyl; m is 0, 1, 2 or 3; and Wherein n is 0, 1, 2 or 3; provided that when R6 is hydrogen, m is 1, 2 or 3. or a salt thereof.

[0019] One embodiment provides a compound of formula (I) or a salt thereof, wherein R1 is -NH2.

[0020] One embodiment provides a compound of formula (I) or a salt thereof, wherein R1 is -NH(CH3).

[0021] One embodiment provides a compound of formula (I) or a salt thereof, wherein each R2 is independently F, Cl, -CN, -CH3, -CH2CH3, -CH(CH3)2, -CH2CH2CH2CH3, -CH2F, -CHF2, -CF3, -OCH3 or cyclopropyl.

[0022] One embodiment provides a compound of formula (I) or a salt thereof, wherein each R2 is independently F, Cl, -CN, -CH3, -CH2CH3, -CH(CH3)2, -CH2CH2CH2CH3, -CH2F, -CHF2 or -CF3.

[0023] One embodiment provides a compound of formula (I) or a salt thereof, wherein each R2 is independently F, Cl, -CN, -CH3, -CH2CH3, -CH2F, -CHF2, or -CF3.

[0024] One embodiment provides a compound of formula (I) or a salt thereof, wherein each R2 is independently F, Cl, -CN, -CH3 or -CF3.

[0025] One embodiment provides a compound of formula (I) or a salt thereof, wherein each R2 is independently -CN, -CH3 or -CF3.

[0026] One embodiment provides a compound of formula (I) or a salt thereof, wherein each R2 is independently F, -CN, -CH3 or -CF3.

[0027] One embodiment provides a compound of formula (I) or a salt thereof, wherein each R2 is independently F, -CN or -CH3.

[0028] One embodiment provides a compound of formula (I) or a salt thereof, wherein n is 0, 1 or 2.

[0029] One embodiment provides a compound of formula (I) or a salt thereof, wherein n is 0 or 1.

[0030] One embodiment provides a compound of formula (I) or a salt thereof, wherein n is 1 or 2.

[0031] One embodiment provides a compound of formula (I) or a salt thereof, wherein n is 0.

[0032] One embodiment provides a compound of formula (I) or a salt thereof, wherein n is 1.

[0033] One embodiment provides a compound of formula (I) or a salt thereof, wherein n is 2.

[0034] One embodiment provides a compound of formula (I) or a salt thereof, wherein each R2 is independently -CN, -CH3, or -CF3; and n is 0, 1, or 2. Included in this embodiment are compounds where n is 1 or 2.

[0035] One embodiment provides a compound of formula (I) or a salt thereof, wherein each R2 is independently -CN, -CH3, or -CF3; and n is 0 or 1. Included in this embodiment are compounds where n is 1.

[0036] One embodiment provides a compound of formula (I) or a salt thereof, wherein each R2 is independently -CN, -CH3, or -CF3; and n is 2.

[0037] One embodiment provides a compound of formula (I) or a salt thereof, wherein each R2 is independently -CN or -CH3; and n is 1 or 2. Included in this embodiment are compounds where n is 1. Also included in this embodiment are compounds where n is 2.

[0038] One embodiment provides a compound of formula (I) or a salt thereof, wherein each R2 is -CH3; and n is 1 or 2. Included in this embodiment are compounds where n is 1. Also included in this embodiment are compounds where n is 2.

[0039] One embodiment provides a compound of formula (I) or a salt thereof, wherein each R4 is independently F, Cl, -CH3, -CH2F, -CHF2, or -CF3.

[0040] One embodiment provides a compound of formula (I) or a salt thereof, wherein each R4 is independently F, -CH3, -CH2F, -CHF2, or -CF3.

[0041] One embodiment provides a compound of formula (I) or a salt thereof, wherein each R4 is independently F, -CH3, -CHF2 or -CF3.

[0042] One embodiment provides a compound of formula (I) or a salt thereof, wherein each R4 is independently F or -CH3.

[0043] One embodiment provides a compound of formula (I) or a salt thereof, wherein each R4 is F3. Included in this embodiment are compounds where m is 1. Also included in this embodiment are compounds where m is 2.

[0044] One embodiment provides a compound of formula (I) or a salt thereof, wherein each R4 is -CH3. Included in this embodiment are compounds where m is 1. Also included in this embodiment are compounds where m is 2.

[0045] One embodiment provides a compound of formula (I) or a salt thereof, wherein m is 0, 1 or 2.

[0046] One embodiment provides a compound of formula (I) or a salt thereof, wherein m is 0 or 1.

[0047] One embodiment provides a compound of formula (I) or a salt thereof, wherein m is 1 or 2.

[0048] One embodiment provides a compound of formula (I) or a salt thereof, wherein m is 1, 2 or 3.

[0049] One embodiment provides a compound of formula (I) or a salt thereof, wherein m is 1.

[0050] One embodiment provides a compound of formula (I) or a salt thereof, wherein R6 is hydrogen; and m is 1, 2 or 3.

[0051] One embodiment provides a compound of formula (I) or a salt thereof, wherein R6 is hydrogen; and m is 1 or 2.

[0052] One embodiment provides a compound of formula (I) or a salt thereof, wherein R6 is hydrogen; and m is 1.

[0053] One embodiment provides a compound of formula (I) or a salt thereof, wherein R6 is hydrogen; and m is 2.

[0054] One embodiment provides a compound of formula (I) or a salt thereof, wherein R6 is hydrogen; and m is 3.

[0055] One embodiment provides a compound of formula (I) or a salt thereof, wherein R6 is hydrogen; m is 1; and R4 is F, Cl, -CH3, -CH2F, -CHF2, or -CF3. Included in this embodiment are compounds where R4 is F, -CH3, -CHF2, or -CF3. Also included in this embodiment are compounds where R4 is F, -CH3, or -CF3. Further included in this embodiment are compounds where R4 is F or -CH3.

[0056] One embodiment provides a compound of formula (I) or a salt thereof, wherein R6 is hydrogen; m is 1; and R4 is F.

[0057] One embodiment provides a compound of formula (I) or a salt thereof, wherein R6 is hydrogen; m is 1; and R4 is -CH3.

[0058] One embodiment provides a compound of formula (I) or a salt thereof, wherein R6 is hydrogen; m is 1 or 2; and R4 is F or -CH3.

[0059] In one embodiment, R6 is C 1-2 Alkyl or C 1-2 The present invention provides a compound of formula (I) or a salt thereof, which is fluoroalkyl.

[0060] In one embodiment, R6 is C 1-2 The present invention provides a compound of formula (I) or a salt thereof, wherein:

[0061] In one embodiment, R6 is C 1-2The present invention provides a compound of formula (I) or a salt thereof, wherein:

[0062] In one embodiment, R6 is C 1-2 The present invention provides a compound of formula (I) or a salt thereof, wherein:

[0063] In one embodiment, R6 is C 1-2 The present invention provides a compound of formula (I) or a salt thereof, wherein

[0064] One embodiment provides a compound of formula (I) or a salt thereof, wherein R6 is -CH3.

[0065] One embodiment provides a compound of formula (I) or a salt thereof, wherein R6 is -CH2F, -CF2H, -CF3 or -CH2CF3.

[0066] One embodiment provides a compound of formula (I) or a salt thereof, wherein R6 is -CH2F, -CF2H or -CF3.

[0067] In one embodiment, R6 is C 1-2 Alkyl or C 1-2 and m is 0; or a salt thereof.

[0068] In one embodiment, R6 is C 1-2 and m is 0; or a salt thereof.

[0069] In one embodiment, R6 is C 1-2 and m is 0; or a salt thereof.

[0070] In one embodiment, R6 is C 1-2 and m is 0; or a salt thereof.

[0071] In one embodiment, R6 is C 1-2 and m is 0; or a salt thereof.

[0072] One embodiment provides a compound of formula (I) or a salt thereof, wherein R6 is -CH3; and m is 0.

[0073] One embodiment provides a compound of formula (I) or a salt thereof, wherein R6 is -CH2F, -CF2H, -CF3 or -CH2CF3; and m is 0.

[0074] One embodiment provides a compound of formula (I) or a salt thereof, wherein R6 is -CH2F, -CF2H or -CF3; and m is 0.

[0075] One embodiment provides a compound of formula (I) or a salt thereof, wherein R1 is -NH2 or -NH(CH3); each R2 is independently -CN, -CH3 or -CF3; each R4 is independently F or -CH3; R6 is hydrogen or -CH3; m is 0 or 1; and n is 1 or 2; with the proviso that when R6 is hydrogen, m is 1.

[0076] One embodiment provides a compound of formula (I) or a salt thereof, wherein R1 is -NH2; each R2 is independently -CN, -CH3 or -CF3; R4 is F or -CH3; R6 is hydrogen or -CH3; m is 0 or 1; and n is 1 or 2; with the proviso that when R6 is hydrogen, m is 1.

[0077] One embodiment provides a compound of formula (I) or a salt thereof, wherein R1 is -NH(CH3); each R2 is independently -CN or -CH3; R4 is F or -CH3; R6 is hydrogen or -CH3; m is 0 or 1; and n is 1 or 2; with the proviso that when R6 is hydrogen, m is 1.

[0078] One embodiment provides a compound of formula (I) or a salt thereof, wherein R1 is -NH2; each R2 is independently -CN, -CH3 or -CF3; each R4 is independently F or -CH3; R6 is -CH3; m is 1; and n is 1 or 2. Included in this embodiment are compounds where each R2 is -CH3.

[0079] One embodiment provides a compound of formula (I) or a salt thereof, wherein R1 is -NH2; each R2 is independently -CN or -CH3; R4 is F or -CH3; R6 is hydrogen; m is 1; and n is 1 or 2.

[0080] One embodiment provides a compound of formula (I) or a salt thereof, wherein R1 is -NH2 or -NH(CH3); each R2 is independently -CH3; R4 is F; R6 is hydrogen or -CH3; m is 0 or 1; and n is 1 or 2; with the proviso that when R6 is hydrogen, m is 1.

[0081] One aspect is a compound having the following structure: [ka] or a salt thereof.

[0082] One aspect is a compound having the following structure: [ka] The present invention provides a compound of formula (I) which is: or a salt thereof.

[0083] One aspect is a compound having the following structure: [ka] The present invention provides a compound of formula (I) which is: or a salt thereof.

[0084] One aspect is a compound having the following structure: [ka] The present invention provides a compound of formula (I) which is: or a salt thereof.

[0085] One aspect is a compound having the following structure: [ka] The present invention provides a compound of formula (I) which is: or a salt thereof.

[0086] One aspect is a compound having the following structure: [ka] The present invention provides a compound of formula (I) which is: or a salt thereof.

[0087] One aspect is a compound having the following structure: [ka] The present invention provides a compound of formula (I) which is: or a salt thereof.

[0088] One aspect is a compound having the following structure: [ka] The present invention provides a compound of formula (I) which is: or a salt thereof.

[0089] One aspect is a compound having the following structure: [ka] The present invention provides a compound of formula (I) which is: or a salt thereof.

[0090] One aspect is a compound having the following structure: [ka] The present invention provides a compound of formula (I) which is: or a salt thereof.

[0091] One aspect is a compound having the following structure: [ka] The present invention provides a compound of formula (I) which is: or a salt thereof.

[0092] One aspect is a compound having the following structure: [ka] The present invention provides a compound of formula (I) which is: or a salt thereof.

[0093] One aspect is a compound having the following structure: [ka] The present invention provides a compound of formula (I) which is: or a salt thereof.

[0094] One embodiment is the compound: 2-amino-6-(2-(2,6-dioxopiperidin-3-yl)-4-methyl-1-oxoisoindolin-5-yl)-4-methylnicotinonitrile (1); 2-amino-6-(2-(2,6-dioxopiperidin-3-yl)-6-methyl-1-oxoisoindolin-5-yl)-4-methylnicotinonitrile (2); 2-amino-6-(2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxoisoindolin-5-yl)-4-methylnicotinonitrile (3);3-(5-(6-amino-4-methylpyridin-2-yl)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione(4);3-(5-(6-amino-4-(trifluoroethyl)pyridin-2-yl)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione(5);3-(4-fluoro-5-(4-methyl-6-(methylamino)pyridin-2-yl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione(6);3-(5 -(6-amino-4,5-dimethylpyridin-2-yl)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (7); 3-((S)-5-(6-amino-4,5-dimethylpyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (8); 3-((R)-5-(6-amino-4,5-dimethylpyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (9); 2-amino-6-(( 3S)-2-(2,6-dioxopiperidin-3-yl)-3-methyl-1-oxoisoindolin-5-yl)-4-methylnicotinonitrile (10); 2-amino-6-((3R)-2-(2,6-dioxopiperidin-3-yl)-3-methyl-1-oxoisoindolin-5-yl)-4-methylnicotinonitrile (11); 3-((S)-5-(4,5-dimethyl-6-(methylamino)pyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (12);3-((R)-5-(4,5-dimethyl-6-(methylamino)pyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (13); 6-((3S)-2-(2,6-dioxopiperidin-3-yl)-3-methyl-1-oxoisoindolin-5-yl)-4-methyl-2-(methylamino)nicotinonitrile (14); 6-((3R)-2-(2,6-dioxopiperidin-3-yl)-3-methyl-1-oxoisoindolin-5-yl)-4-methyl-2-(methylamino)nicotinonitrile (15);3-((S)-5-(6-amino-4,5-dimethylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (16);3-((R)-5-(6-amino-4,5-dimethylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (17);3-(5-(6-amino-3-methylpyridin-2-yl)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (18);3-(5 -(6-amino-5-fluoro-4-methylpyridin-2-yl)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (19); 3-(5-(6-amino-3-fluoro-4-methylpyridin-2-yl)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (20); 3-(5-(6-amino-5-fluoro-4-methylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (21-22); 2-amino-6-( (3R)-2-(2,6-dioxopiperidin-3-yl)-4-fluoro-3-methyl-1-oxoisoindolin-5-yl)-4-methylnicotinonitrile (23); 3-((R)-5-(6-amino-4-methylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (24); 3-((R)-5-(6-amino-4-(trifluoromethyl)pyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (25);3-((R)-5-(6-amino-3-methylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (26); 3-((R)-5-(6-aminopyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (27); (R)-3-((R)-5-(6-amino-4,5-dimethylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2 3-((R)-5-(6-amino-5-fluoro-4-methylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (28); 3-((R)-5-(6-amino-5-fluoro-4-methylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (29); or 3-((S)-5-(6-amino-5-fluoro-4-methylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (30), or a salt thereof.

[0095] One embodiment provides a compound of formula (I) or a salt thereof, wherein the compound is 3-((R)-5-(6-amino-5-fluoro-4-methylpyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione.

[0096] One embodiment provides a compound of formula (I) or a salt thereof, wherein the compound is 3-((S)-5-(6-amino-5-fluoro-4-methylpyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione.

[0097] The present invention may be embodied in other specific forms without departing from its spirit or essential characteristics. The present invention encompasses any and all combinations of the aspects and / or embodiments of the present invention described herein. It is understood that any and all embodiments of the present invention may be combined with any and all other embodiments to describe further embodiments. It is also understood that individual elements of the embodiments are meant to be combined with any and all other elements from any and all embodiments to describe further embodiments.

[0098] The features and advantages of the present invention may be more readily understood by those skilled in the art upon reading the following detailed description. It is understood that, for clarity, certain features of the invention that are described in the context of another embodiment may be combined to form a single embodiment. Conversely, various features of the invention that are described in the context of a single embodiment for brevity may be combined to form subcombinations thereof. The embodiments identified herein as exemplary or preferred are intended to be illustrative and not limiting.

[0099] Unless otherwise specified herein, words referred to in the singular may also include the plural. For example, "a" and "an" can refer to either "one" or "one or more."

[0100] As used herein, the phrase "compound and / or salt thereof" refers to at least one compound, a salt of at least one compound, or a combination thereof. For example, a compound of formula (I) and / or a salt thereof includes one compound of formula (I); two compounds of formula (I); a salt of one compound of formula (I); one compound of formula (I) and one or more salts of compounds of formula (I); and a salt of two or more compounds of formula (I).

[0101] Unless otherwise specified, any atom with unsatisfied valences is assumed to include enough hydrogen atoms to satisfy the valences.

[0102] The definitions set forth herein take precedence over definitions set forth in any patents, patent applications, and / or published patent applications incorporated herein by reference.

[0103] Listed below are definitions of various terms used to describe this invention. These definitions apply to the terms as they are used throughout the specification, either individually (unless otherwise limited in specific instances) or as part of a larger group.

[0104] Throughout the specification, groups and substituents may be chosen by one skilled in the art to provide stable moieties and compounds.

[0105] According to the practice used in the art, [ka] is used in structural formulas herein to represent the bond that is the point of attachment of a moiety or substituent to a core or backbone structure.

[0106] The terms "halo" and "halogen" as used herein refer to F, Cl, Br, and I.

[0107] The term "cyano" refers to the group --CN.

[0108] The term "amino" refers to the group -NH2.

[0109] The term "oxo" refers to the group =O.

[0110] The term "alkyl" as used herein refers to both branched and straight chain saturated aliphatic hydrocarbon groups, for example, having 1 to 12 carbon atoms, 1 to 6 carbon atoms, and 1 to 4 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (e.g., n-propyl and i-propyl), butyl (e.g., n-butyl, i-butyl, sec-butyl, and t-butyl), and pentyl (e.g., n-pentyl, isopentyl, neopentyl), n-hexyl, 2-methylpentyl, 2-ethylbutyl, 3-methylpentyl, and 4-methylpentyl. When a number appears in subscript after the symbol "C," the subscript more specifically qualifies the number of carbon atoms that a particular group may contain. For example, "C 1-4 "Alkyl" means straight and branched chain alkyl groups having 1 to 4 carbon atoms.

[0111] As used herein, the term "fluoroalkyl" is intended to include both branched and straight-chain saturated aliphatic hydrocarbon groups substituted with one or more fluorine atoms. For example, 1-4 "Fluoroalkyl" is meant to include C1, C2, C3, and C4 alkyl groups substituted with one or more fluorine atoms. Representative examples of fluoroalkyl groups include, but are not limited to, -CF3 and -CH2CF3.

[0112] The present invention is intended to include all isotopes of atoms contained in the compounds of the present invention. Isotopes include atoms having the same atomic number but different mass numbers. By way of general example and without limitation, isotopes of hydrogen include deuterium (D) and tritium (T). Isotopes of carbon include 13 C and 14 C. Isotopically labeled compounds of the invention can generally be prepared by conventional techniques known to those of skill in the art, or by methods analogous to those described herein, substituting the appropriate isotopically labeled reagent for the non-labeled reagent otherwise used.

[0113] As used herein, the phrase "pharmacologically acceptable" refers to compounds, substances, compositions, and / or dosage forms that are suitable, within the scope of ordinary medical judgment, for contact with the tissues of human beings and animals without undue toxicity, irritation, allergic response, or other problem or complication and that afford a reasonable benefit / risk ratio.

[0114] Compounds of formula (I) may form salts, which are also within the scope of the present invention. Unless otherwise specified, a reference to a compound of the invention is understood to include a reference to one or more salts thereof. The term "salt" refers to acid and / or base salts formed with inorganic and / or organic acids and bases. In addition, the term "salt" may include zwitterions (internal salts), for example, when a compound of formula (I) has both a basic moiety (e.g., an amine or a pyridine or imidazole ring) and an acidic moiety (e.g., a carboxylic acid). Pharmaceutically acceptable (i.e., non-toxic and physiologically acceptable) salts are preferred, for example, acceptable metal salts and amine salts, in which the cation does not significantly contribute to the toxicity or biological activity of the salt. However, other salts may also be useful, for example, in isolation or purification steps that may be used in the preparation process, and therefore other salts are considered to be within the scope of the present invention. Salts of compounds of formula (I) may be formed, for example, by reacting a compound of formula (I) with a certain amount of acid or base (e.g., one equivalent) and precipitating the salt in a solvent, for example, or from an aqueous solution followed by lyophilization.

[0115] Examples of acid addition salts include acetates (e.g., acetates prepared from acetic acid or trihaloacetic acids (e.g., trifluoroacetic acid)), adipates, alginates, ascorbates, aspartates, benzoates, benzenesulfonates, hydrogensulfates, borates, butyrates, citrates, camphorates, camphorsulfonates, cyclopentanepropionates, digluconates, dodecylsulfates, ethanesulfonates, fumarates, glucoheptanoates, glycerophosphates, hemisulfates, heptanoates, hexanoates, hydrochlorides (prepared from hydrochloric acid), hydrobromides (prepared from hydrobromide), and the like. salts such as those described herein), hydroiodides, maleates (prepared from maleic acid), 2-hydroxyethanesulfonates, lactates, methanesulfonates (prepared from methanesulfonic acid), 2-naphthalenesulfonates, nicotinates, nitrates, oxalates, pectinates, persulfates, 3-phenylpropionates, phosphates, picrates, pivalates, propionates, salicylates, succinates, sulfates (e.g., prepared from sulfuric acid), sulfonates (e.g., those described herein), tartrates, thiocyanates, toluenesulfonates (e.g., tosylates), undecanoates, and the like.

[0116] Examples of base salts include ammonium salts, alkali metal salts (e.g., sodium, lithium, and potassium salts), alkaline earth metal salts (e.g., calcium and magnesium salts), barium, zinc, and aluminum salts, organic base salts such as organic amines (e.g., trialkylamines (e.g., triethylamine), procaine, dibenzylamine, N-benzyl-β-phenethylamine, 1-ephenamine, N,N'-dibenzylethylene-diamine, dehydroabietylamine, N-ethylpiperidine, benzylamine, dicyclohexylamine), or similar pharma- ceutically acceptable amines, and amino acid salts (e.g., arginine, lysine), and the like. Basic nitrogen-containing groups may be quaternized with reagents such as lower alkyl halides (e.g., methyl, ethyl, propyl, and butyl chlorides, bromides, and iodides), dialkyl sulfates (e.g., methyl, ethyl, propyl, and butyl chlorides, bromides, and iodides), longer chain halides (e.g., decyl, lauryl, myristyl, and stearyl chlorides, bromides, and iodides), aralkyl halides (e.g., benzyl and phenethyl bromides), and other groups.

[0117] The compound of formula (I) may be provided as an amorphous solid or a crystalline solid. The compound of formula (I) may be provided as a solid by lyophilization.

[0118] In addition, solvates (e.g., hydrates) of the compounds of formula (I) should be considered within the scope of the present invention. The term "solvate" refers to a physical association of a compound of formula (I) with one or more organic or inorganic solvent molecules. This physical association includes hydrogen bonding. In some cases, it is possible to isolate the solvate, for example when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid. "Solvate" includes both solution-phase and isolable solvates. Examples of solvates include hydrates, ethanolates, methanolates, isopropanolates, acetonitrile solvates, and ethyl acetate solvates. Methods of solvation are known in the art.

[0119] Various forms of prodrugs are well known in the art and are described in Rautio, J. et al., Nature Review Drug Discovery, 17, 559-587 (2018).

[0120] In addition, the compounds of formula (I) may be prepared, isolated, purified, and obtained as compositions containing 99% or greater of the compound of formula (I) ("substantially pure"), which are then used or formulated as described herein. Such "substantially pure" compounds of formula (I) are also considered herein to be part of the invention.

[0121] By "stable compound" and "stable structure" is intended a compound that is sufficiently robust that it will not decompose upon isolation to a useful degree of purity from a reaction mixture and formulation into an efficacious therapeutic agent. The present invention embodies stable compounds.

[0122] The term "Helios inhibitor" refers to an agent that reduces Helios protein levels and Helios activity levels in cells and / or suppresses Helios expression levels to regulate Treg differentiation. Helios inhibitors can be reversible or irreversible inhibitors.

[0123] As used herein, "Helios" protein refers to a protein that is a member of the Ikaros family of zinc finger proteins. In humans, Helios is encoded by the IKZF2 gene. Helios is also known as Ikaros family zinc finger 2, ANF1A2, ZNF1A2, ZNFN1A2, zinc finger proteins, subfamilies 1A, 2, and Ikaros family zinc finger protein 2. Members of this protein family include Ikaros, Helios, Aiolos, Eos, and Pegasus. As used herein, Helios protein includes various isoforms, including isoforms 1-5 listed below. [Table 1] [Table 2] [Table 3] [Table 4] [Table 5]

[0124] The "Helios" isoforms 1, 2, 4, 6, and 7 contain a degron, FHCNQCGASFTQKGNLLRHIKLH (SEQ ID NO: 6) (bold underlined). A degron is a part of a protein that plays a role in regulating the rate of protein degradation.

[0125] As used herein, the "Eos" protein is encoded by the IKZF4 gene and is also known as Ikaros family zinc finger 4, ZNFN1A4, zinc finger protein, subfamily 1A, 4, Ikaros family zinc finger protein 4, and KIAA1782. The "Eos" protein includes isoforms encoded by the following two human isoforms 1 (Q9H2S9-1) and 2 (Q9H2S9-2). [Table 6] [Table 7]

[0126] The "Eos" protein isoforms 1 and 2 contain a degron, FHCNQCGASFTQKGNLLRHIKLH (SEQ ID NO: 6) (bold underlined portion), which is identical to the degron of the "Helios" protein.

[0127] As used herein, the "Ikaros" protein is encoded by the IKZF1 gene. Ikaros is also known as Ikaros family zinc finger 1, ZNFN1A1, zinc finger protein, subfamily 1A, 1, Ikaros family zinc finger protein 1, IK1, lymphoid transcription factor LyF-1, Hs.54452, PPP1R92, protein phosphatase 1, regulatory subunit 92, PRO0758, CVID13, and CLL-associated antigen KW-6. Ikaros proteins include isoforms encoded by amino acid sequences Q13422-1, Q13422-2, Q13422-3, Q13422-4, Q13422-7, and Q13422-8. Ikaros proteins also include isoforms encoded by amino acid sequences Q13422-5 and Q13422-6.

[0128] As used herein, the "Aiolos" protein is encoded by the IKZF3 gene. Aiolos protein is also known as Ikaros family zinc finger 3, ZNFN1A3, zinc finger protein, subfamily 1A, 3, Ikaros family zinc finger protein 3, and AIO. Aiolos protein includes isoforms encoded by the amino acid sequences Q9UKT9-1, Q9UKT9-3, Q9UKT9-4, Q9UKT9-6, Q9UKT9-7, Q9UKT9-8, Q9UKT9-9, and Q9UKT9-14. Aiolos proteins also include isoforms encoded by the amino acid sequences Q9UKT9-2, Q9UKT9-5, Q9UKT9-10, Q9UKT9-11, Q9UKT9-12, and Q9UKT9-13, Q9UKT9-15, and Q9UKT9-16.

[0129] As used herein, the "Pegasus" protein is also known as Ikaros family zinc finger 5, ZNFN1A5, zinc finger protein, subfamily 1A, 5, and Ikaros family zinc finger protein 5. Pegasus is encoded by the IKZF5 gene.

[0130] As used herein, the term "contacting" refers to bringing together a specified moiety in vitro or in vivo. For example, "contacting" a Helios protein with a compound of formula (I) includes administering a compound of the present invention to an individual or patient (e.g., a human) having a Helios protein, as well as introducing a compound of formula (I) into, for example, a sample containing cells or a purification product containing a Helios protein.

[0131] As used herein, the terms "treat" and "treatment" refer to any intervention, method, or administration of an active agent to a subject for the purpose of ameliorating, alleviating, ameliorating, inhibiting, delaying, or suppressing the progression, onset, severity, or recurrence of a symptom, complication, condition, or biochemical manifestation of a disease. In contrast, "prophylaxis" or "prevention" refers to administration to a non-affected subject to prevent the onset of a disease. "Treat" and "treatment" do not include prophylaxis or prevention.

[0132] "Therapeutically effective amount" is intended to include an amount of a compound of the invention alone, or in combination with a claimed compound, or in combination with other active ingredients, effective to reduce Helios protein levels and Helios activity levels in a cell and / or inhibit Helios expression levels, or to treat or prevent viral infections and proliferative diseases (e.g., cancer).

[0133] The term "cell" as used herein is intended to refer to in vitro, ex vivo or in vivo cells. In some embodiments, ex vivo cells can be part of tissue samples taken from organisms (e.g., mammals). In some embodiments, in vitro cells can be cells in cell culture. In some embodiments, in vivo cells are living cells in organisms (e.g., mammals).

[0134] The term "patient" includes human and other mammalian subjects receiving either therapeutic or prophylactic treatment.

[0135] The term "subject" includes any animal, human or non-human. For example, the methods and compositions disclosed herein are used to treat subjects suffering from cancer. Non-human animals include any vertebrate, for example, mammals and non-mammals (non-human primates, sheep, dogs, cows, birds, amphibians, reptiles, etc.). In some embodiments, the subject is a human.

[0136] As used herein, the phrase "pharmaceutical acceptable carrier" refers to a pharma- ceutically acceptable substance, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, processing aid (e.g., lubricant, magnesium talc, calcium or zinc stearate, or stearic acid), or solvent encapsulant, which is involved in the transport or delivery of a particular compound 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 in the formulation (i.e., including adjuvants, excipients, or vehicles (e.g., diluents, preservatives, fillers, flow regulators, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, fragrances, antibacterial agents, antifungal agents, lubricants, and dispensing agents) depending on the nature of the method of administration and the dosage form) and not deleterious to the patient.

[0137] The term "pharmaceutical composition" means a composition comprising a compound of the present invention in combination with at least one additional pharma- ceutically acceptable carrier.

[0138] (usefulness) The compounds of formula (I) are useful in the treatment of cancer.

[0139] In one embodiment, the present invention provides a pharmaceutical combination of a compound of formula (I) and / or a pharma- ceutical acceptable salt thereof, a stereoisomer thereof or a tautomer thereof with another therapeutic agent for simultaneous, separate or sequential use in the treatment and / or prevention of diseases or disorders associated with the activity of Helios protein, which is used to reduce Helios protein levels, Helios activity levels and / or Helios expression levels in cells to modulate the differentiation of Tregs.

[0140] The compounds of formula (I) and pharmaceutical compositions comprising at least one compound of formula (I) are useful for treating or preventing any disease or condition associated with the activity of Helios protein. Such diseases or conditions include viral and other infectious diseases (e.g., skin infections, GI infections, urinary tract infections, urogenital infections, systemic infections), and proliferative diseases (e.g., cancer). The compounds of formula (I) and pharmaceutical compositions comprising at least one compound of formula (I) can be administered to animals, preferably mammals (e.g., laboratory animals, cats, dogs, mice, rats), and more preferably humans. Any method of administration can be used to deliver the compound or pharmaceutical composition to the patient. In some embodiments, the compounds of formula (I) or pharmaceutical compositions comprising at least one compound of formula (I) are administered orally. In other embodiments, the pharmaceutical compositions comprising formula (I) or at least one compound of formula (I) are administered parenterally.

[0141] The compound of formula (I) can selectively reduce Helios protein levels and Helios activity levels and / or suppress Helios expression levels in cells to control Treg differentiation. For example, the compound of formula (I) can be used to reduce Helios activity levels and / or inhibit Helios expression levels in subjects or cells that need to selectively reduce Helios protein levels and Helios activity levels and / or suppress Helios expression levels to control Treg differentiation, and the compound of formula (I) or a salt thereof can be administered in an inhibitory amount.

[0142] In some embodiments, the compound of formula (I) is administered sequentially prior to administration of the immuno-oncology agent. In other embodiments, the compound of formula (I) is administered simultaneously with the immuno-oncology agent. In yet other embodiments, the compound of formula (I) is administered sequentially following administration of the immuno-oncology agent.

[0143] In another embodiment, the compound of formula (I) may be formulated with an immuno-oncology agent.

[0144] The immuno-oncology agent includes, for example, small molecule drugs, antibodies, or other biological molecules. Examples of biological immuno-oncology agents include, but are not limited to, cancer vaccines, antibodies, and cytokines. In one embodiment, the antibody is a monoclonal antibody. In another embodiment, the monoclonal antibody is a humanized antibody or a human antibody.

[0145] In some embodiments, the immuno-oncology agent is either (i) an agonist of a stimulatory receptor (including costimulatory), or (ii) an antagonist of an inhibitory signal (including co-inhibitory) on a T cell, both of which result in amplification of an antigen-specific T cell response (often referred to as an immune checkpoint regulator).

[0146] Certain stimulatory and inhibitory molecules belong to the immunoglobulin superfamily (IgSF). One important family of membrane-bound ligands that bind to costimulatory or co-inhibitory receptors is the B7 family, which includes B7-1, B7-2, B7-H1 (PD-L1), B7-DC (PD-L2), B7-H2 (ICOS-L), B7-H3, B7-H4, B7-H5 (VISTA), and B7-H6. Another family of membrane-bound ligands that bind to costimulatory or coinhibitory receptors are the TNF family molecules that bind to the cognate TNF receptor family, including CD40 and CD40L, OX-40, OX-40L, CD70, CD27L, CD30, CD30L, 4-1BBL, CD137(4-1BB), TRAIL / Apo2-L, TRAILR1 / DR4, TRAILR2 / DR5, TRAILR3, TRAILR4, OPG, RANK, RANKL, TWEAKR / Fn14, TWEAK, BAFFR, EDAR, XEDAR, TACI, April, BCMA, LTβR, LIGHT, DcR3, HVEM, VEGI / TL1A, TRAMP / DR3, E Includes DAR, EDA1, XEDAR, EDA2, TNFR1, lymphotoxin α / TNFβ, TNFR2, TNFα, LTβR, lymphotoxin α1β2, FAS, FASL, RELT, DR6, TROY, and NGFR.

[0147] In certain embodiments, T cell responses may be stimulated by a combination of a compound of formula (I) of the present invention and one or more of the following: (i) antagonists of proteins that inhibit T cell activation (e.g., immune checkpoint inhibitors), such as CTLA-4, PD-1, PD-L1, PD-L2, LAG-3, TIM-3, Galectin 9, CEACAM-1, BTLA, CD69, Galectin 1, TIGIT, CD113, GPR56, VISTA, 2B4, CD48, GARP, PD1H, LAIR1, TIM-1, and TIM-4, and (ii) agonists of proteins that stimulate T cell activation, such as B7-1, B7-2, CD28, 4-1BB (CD137), 4-1BBL, ICOS, ICOS-L, OX40, OX40L, GITR, GITRL, CD70, CD27, CD40, DR3, and CD28H.

[0148] For cancer treatment, other drugs that can be combined with the compound of formula (I) can include antagonists of inhibitory receptors on NK cells, or agonists of activating receptors on NK cells. For example, the compound of formula (I) can be combined with antagonists of KIR, such as lirilumab.

[0149] Further agents for use in combination therapy include agents that inhibit or deplete macrophages or monocytes, including, but are not limited to, CSF-1R antagonists, such as RG7155 (WO11 / 70024, WO11 / 107553, WO11 / 131407, WO13 / 87699, WO13 / 119716, WO13 / 132044) or CSF-1R antagonist antibodies, including FPA-008 (WO11 / 140249; WO13169264; WO14 / 036357).

[0150] In another embodiment, the compounds of formula (I) may be used with one or more of agonistic agents that bind positive co-stimulatory receptors, blocking agents that attenuate signaling through inhibitory receptors, antagonists, and one or more agents that increase the frequency of anti-tumor T cells systemically, agents that overcome different immune suppressive pathways in the tumor microenvironment (e.g., blocking inhibitory receptor engagement (e.g., PD-L1 / PD-1 interaction), depleting or inhibiting Treg cells (e.g., using anti-CD25 monoclonal antibodies (e.g., daclizumab) or by depletion with anti-CD25 beads in vitro), inhibiting metabolic enzymes such as IDO, or restoring / preventing T cell anergy or T cell depletion), and agents that cause innate immune activation and / or inflammation in the tumor area.

[0151] In some embodiments, the immuno-oncology agent is a CTLA-4 antagonist, such as an antagonistic CTLA-4 antibody. Suitable CTLA-4 antibodies include, for example, Yervoy (ipilimumab), or tremelimumab.

[0152] In another embodiment, the immuno-oncology agent is a PD-1 antagonist, for example, an antagonistic PD-1 antibody. Suitable PD-1 antibodies include, for example, Opdivo (nivolumab), Keytruda (pembrolizumab), or MEDI-0680 (AMP-514; WO2012 / 145493). The immuno-oncology agent may also include pidilizumab (CT-011), although its specificity for PD-1 binding has been questioned. Another approach to target the PD-1 receptor is a recombinant protein consisting of the extracellular domain of PD-L2 (B7-DC) fused to the Fc portion of IgG1, called AMP-224.

[0153] In another embodiment, the immuno-oncology agent includes a PD-L1 antagonist, such as an antagonistic PD-L1 antibody. Suitable PD-L1 antibodies include, for example, MPDL3280A (RG7446; WO2010 / 077634), durvalumab (MEDI4736), BMS-936559 (WO207 / 005874), and MSB0010718C (WO2013 / 79174).

[0154] In another embodiment, the immuno-oncology agent is a LAG-3 antagonist, such as an antagonistic LAG-3 antibody.Suitable LAG3 antibodies include, for example, BMS-986016 (WO10 / 19570, WO14 / 08218), or IMP-731 or IMP-321 (WO08 / 132601, WO09 / 44273).

[0155] In another embodiment, the immuno-oncology agent is a CD137 (4-1BB) agonist, such as an agonistic CD137 antibody. Suitable CD137 antibodies include, for example, urelumab and PF-05082566 (WO12 / 32433).

[0156] In another embodiment, the immuno-oncology agent is a GITR agonist, for example, an agonistic GITR antibody. Suitable CD137 antibodies include, for example, BMS-986153, BMS-986156, TRX-518 (WO06 / 105021, WO09 / 009116) and MK-4166 (WO11 / 028683).

[0157] In another embodiment, the immuno-oncology agent is an IDO antagonist.Suitable IDO antagonists include, for example, INCB-024360 (WO206 / 122150, WO07 / 75598, WO08 / 36653, WO08 / 36642), indoximod or NLG-919 (WO09 / 73620, WO09 / 1156652, WO11 / 56652, WO12 / 142237).

[0158] In another embodiment, the immuno-oncology agent is an OX40 agonist, such as an agonistic OX40 antibody. Suitable OX40 antibodies include, for example, MEDI-6383 or MEDI-6469.

[0159] In another embodiment, the immuno-oncology agent is an OX40L antagonist, such as an antagonistic OX40 antibody. Suitable OX40L antagonists include, for example, RG-7888 (WO06 / 029879).

[0160] In another embodiment, the immuno-oncology agent is a CD40 agonist, such as an agonistic CD40 antibody.In yet another embodiment, the immuno-oncology agent is a CD40 antagonist, such as an antagonistic CD40 antibody.Suitable CD40 antibodies include, for example, lucatumumab or dacetuzumab.

[0161] In another embodiment, the immuno-oncology agent is a CD27 agonist, such as an antagonistic CD27 antibody. Suitable CD27 antibodies include, for example, varlilumab.

[0162] In another embodiment, the immuno-oncology agent (against B7H3) is MGA271 (WO11 / 109400).

[0163] Combination therapy is intended to include administration of the therapeutic agents in a sequential manner, i.e., each therapeutic agent is administered at various different times, as well as administration of the therapeutic agents or at least two therapeutic agents in a substantially simultaneous manner. Substantially simultaneous administration can be achieved, for example, by administering to the patient a single dosage form of each therapeutic agent with a fixed ratio, or a single dosage form of each therapeutic agent. Sequential or substantially simultaneous administration of each therapeutic agent can be performed by any suitable route, including, but not limited to, oral, intravenous, intramuscular, and direct absorption through mucosal membranes. The therapeutic agents can be administered by the same route or by different routes. For example, a first therapeutic agent in a selected combination can be administered by intravenous injection, while another therapeutic agent in the combination can be administered orally. Alternatively, for example, all therapeutic agents can be administered orally, or all therapeutic agents can be administered by intravenous injection. The administration of the therapeutic agents described above can also be combined with other biologically active ingredients and non-pharmaceutical therapies (e.g., surgery or radiation therapy) to perform combination therapy. When the combination therapy further includes a non-drug treatment, the non-drug treatment can be administered at any suitable time, so long as the beneficial effect resulting from the synergistic action of the combination of the therapeutic agent and the non-drug treatment is achieved, e.g., in suitable cases, the beneficial effect is achieved even when the non-drug treatment is temporarily suspended, perhaps for days or weeks, from administration of the therapeutic agent.

[0164] The types of cancer that may be treated with the compounds of formula (I) include, but are not limited to, brain cancer, skin cancer, bladder cancer, ovarian cancer, breast cancer, stomach cancer, pancreatic cancer, prostate cancer, colon cancer, blood cancer, lung cancer and bone cancer. Examples of such cancer types include neuroblastoma, intestinal cancer (e.g., rectal cancer, colon cancer, familial adenomatous polyposis and hereditary nonpolyposis colorectal cancer), esophageal cancer, lip cancer, laryngeal cancer, hypopharyngeal cancer, tongue cancer, salivary gland cancer, stomach cancer, adenocarcinoma, medullary thyroid cancer, papillary thyroid cancer, kidney cancer, renal parenchymal cancer, ovarian cancer, cervical cancer, uterine cancer, endometrial cancer, choriocarcinoma, pancreatic cancer, prostate cancer, testicular cancer, breast cancer, urinary tract cancer, melanoma, brain tumors (e.g., glioblastoma, astrocytoma, meningioma, medulloblastoma and peripheral neuroectodermal tumors), Hodgkin's lymphoma, and tumors, non-Hodgkin's lymphoma, Burkitt's lymphoma, acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), adult T-cell leukemia-lymphoma, diffuse large B-cell lymphoma (DLBCL), hepatocellular carcinoma, gallbladder carcinoma, bronchial carcinoma, small cell lung cancer, non-small cell lung cancer, multiple myeloma, basal cell carcinoma, teratoma, retinoblastoma, choroidal melanoma, seminoma, rhabdomyosarcoma, craniopharyngioma, osteosarcoma, chondrosarcoma, myosarcoma, liposarcoma, fibrosarcoma, Ewing's sarcoma and plasmacytoma.

[0165] One or more additional pharmaceutical agents or therapeutic methods (e.g., antiviral agents, chemotherapeutic or other anti-cancer agents, immune enhancers, immunosuppressants, radiation, antitumor and antiviral vaccines, cytokine therapy (e.g., IL2 and GM-CSF), and / or tyrosine kinase inhibitors) may be used in combination with the compounds of formula (I) as appropriate to treat a disease, disorder, or condition related to a Helios protein. The agents may be combined with the compounds in a single dosage form, or the agents may be administered simultaneously or sequentially in different dosage forms.

[0166] Suitable chemotherapeutic or other anti-cancer agents include, for example, alkylating agents (including but not limited to, nitrogen mustards, ethylenimine derivatives, alkylsulfonic acids, nitrosoureas, and triazenes), such as uracil mustard, chlormethine, cyclophosphamide (Cytoxan®), ifosfamide, melphalan, chlorambucil, pipobroman, triethylene-melamine, triethylenethiophosphoramine, busulfan, carmustine, lomustine, streptozocin, dacarbazine, and temozolomide.

[0167] In the treatment of melanoma, suitable agents for use in combination with the compounds of formula (I) include dacarbazine (DTIC), optionally with other chemotherapeutic agents (e.g., carmustine (BCNU) and cisplatin); the "Dartmouth regimen" consisting of DTIC, BCNU, cisplatin and tamoxifen; cisplatin, vinblastine, and DTIC, temozolomide or Yervoy. TM The compounds of formula (I) may also be combined with immunotherapeutic agents, such as cytokines, such as interferon alpha, interleukin 2, and tumor necrosis factor (TNF), in the treatment of melanoma.

[0168] The compounds of formula (I) can also be used in combination with vaccine therapy in the treatment of melanoma.Anti-melanoma vaccines are similar in some respects to anti-viral vaccines used to prevent diseases caused by viruses (e.g., polio, measles, and mumps).Attenuated melanoma cells or parts of melanoma cells called antigens can be injected into patients to stimulate the body's immune system to destroy melanoma cells.

[0169] Melanomas confined to the arms or legs may also be treated using hyperthermic perfusion therapy with a combination of drugs including one or more compounds of formula (I). This treatment protocol involves temporarily isolating the circulatory system of the affected limb from the rest of the body and injecting high concentrations of chemotherapy drugs into the arteries of the affected limb, delivering high doses to the tumor site that would cause serious side effects if exposed to internal organs. Typically, this treatment involves warming bodily fluids to 38.9°C to 40°C. Melphalan is the drug most frequently used in this chemotherapy. Another drug called tumor necrosis factor (TNF) may also be used.

[0170] Suitable chemotherapeutic or other anti-cancer agents include, for example, antimetabolites (including but not limited to folate antagonists, pyrimidine analogs, purine analogs, and adenosine deaminase inhibitors), such as methotrexate, 5-fluorouracil, floxuridine, cytarabine, 6-mercaptopurine, 6-thioguanine, fludarabine phosphate, pentostatin, and gemcitabine.

[0171] Suitable chemotherapeutic or other anti-cancer agents further include, for example, certain natural products and their derivatives (e.g., vinca alkaloids, antitumor antibiotics, enzymes, lymphokines and epipodophyllotoxins), such as vinblastine, vincristine, vindesine, bleomycin, dactinomycin, daunorubicin, doxorubicin, epirubicin, idarubicin, cytarabine, paclitaxel (taxol), mithramycin, deoxycoformycin, mitomycin C, L-asparaginase, interferons (especially IFNα), etoposide, and teniposide.

[0172] Other cytotoxic agents include navelbine, CPT-11, anastrozole, letrozole, capecitabine, raloxifene, and droloxifene.

[0173] Suitable cytotoxic agents also include, for example, epipodophyllotoxins; anti-neoplastic enzymes; topoisomerase inhibitors; procarbazine; mitoxantrone; platinum coordination complexes (e.g., cisplatin and carboplatin); biological response modifiers; growth inhibitory agents; anti-hormonal therapeutics; leucovorin; tegafur; and hematopoietic growth factors.

[0174] Other anti-cancer agents include antibody drugs such as trastuzumab (Herceptin®), antibodies against costimulatory molecules (such as CTLA-4, 4-1BB and PD-1), or antibodies against cytokines (IL-1O or TGF-β).

[0175] Other anti-cancer agents also include those that block immune cell migration, such as antagonists to chemokine receptors (eg, CCR2 and CCR4).

[0176] Other anti-cancer drugs also include anti-cancer drugs that enhance the immune system, such as adjuvants or adoptive T-cell transfer.

[0177] Anti-cancer vaccines include dendritic cell vaccines, synthetic peptide vaccines, DNA vaccines and recombinant viral vaccines.

[0178] The pharmaceutical compositions of the present invention may optionally contain at least one signal transduction inhibitor (STI). A "signal transduction inhibitor" is an agent that selectively inhibits one or more critical steps in a signal transduction pathway in the normal function of cancer cells, thereby inducing apoptosis. Suitable STIs include, but are not limited to, (i) bcr / abl kinase inhibitors (e.g., STI 571 (GLEEVEC®)); (ii) epidermal growth factor (EGF) receptor inhibitors (e.g., kinase inhibitors (IRESSA®, SSI-774)) and antibodies (Imclone: ​​C225 [Goldstein et al., Clin. Cancer Res., 1:1311-1318 (1995)], and Abgenix: ABX-EGF); (iii) her-2 / neu receptor inhibitors (e.g., farnesyltransferase inhibitors (FTIs) (e.g., L-744,832 [Kohl et al., Nat. Med., 1(8):792-797 (1995)]); (iv) Akt family kinase or Akt pathway inhibitors (e.g., rapamycin (e.g., Sekulic et al., Cancer Res., 60:3504-3513 (200)); (v) cell cycle kinase inhibitors (e.g., flavopiridol and UCN-O1 (see, e.g., Curr. Med. Chem. Anti-Canc. Agents, 3:47-56 (203)); and (vi) phosphatidylinositol kinase inhibitors (e.g., LY294002 (see, e.g., Vlahos et al., J. Biol. Chem., 269:5241-5248 (204)); (1994)). Alternatively, at least one STI and at least one compound of formula (I) can be formulated in separate pharmaceutical compositions. In certain embodiments of the present invention, at least one compound of formula (I) and at least one STI can be administered to a patient simultaneously or sequentially. In other words, at least one compound of formula (I) can be administered first, at least one STI can be administered first, or at least one compound of formula (I) and at least one STI can be administered simultaneously.Furthermore, when more than one compound of formula (I) and / or an STI is used, the compounds may be administered in any order.

[0179] The present invention further provides pharmaceutical compositions comprising at least one compound of formula (I), optionally at least one chemotherapeutic agent, and optionally at least one antiviral agent, in a pharma- ceutically acceptable carrier for treating a chronic viral infection in a patient.

[0180] Also provided is a method of treating a chronic viral infection in a patient by administering an effective amount of the pharmaceutical composition.

[0181] In certain embodiments of the present invention, at least one compound of formula (I) and at least one chemotherapeutic agent are administered to patient simultaneously or sequentially.In other words, at least one compound of formula (I) can be administered first, at least one chemotherapeutic agent can be administered first, or at least one compound of formula (I) and at least one chemotherapeutic agent can be administered simultaneously.In addition, when more than one compound of formula (I) and / or chemotherapeutic agent are used, the compounds can be administered in any order.Similarly, any antiviral drug or STI can be administered at any time compared to the administration of the compound of formula (I).

[0182] Chronic viral infections that may be treated using the combination therapy include, but are not limited to, diseases caused by Hepatitis C virus (HCV), human papillomavirus (HPV), cytomegalovirus (CMV), herpes simplex virus (HSV), Epstein-Barr virus (EBV), varicella zoster virus, coxsackievirus, and human immunodeficiency virus (HIV).

[0183] Suitable antiviral agents contemplated for use in combination with the compounds of formula (I) may include nucleoside and nucleotide reverse transcriptase inhibitors (NRTIs), non-nucleoside reverse transcriptase inhibitors (NNRTIs), protease inhibitors and other antiviral agents.

[0184] Examples of suitable NRTIs include zidovudine (AZT); didanosine (ddl); zalcitabine (ddC); stavudine (d4T); lamivudine (3TC); abacavir (1592U89); adefovir pivoxil [bis(POM)-PMEA]; lobucavir (BMS-180194); BCH-I0652; emtricitabine [(-)-FTC]; β-L-FD4 (also called β-L-D4C, which has the name β-L-2',3'-dideoxy-5-fluoro-cytidine); DAPD, ((-)-β-D-2,6-diamino-purine dioxolane); and rhodenosine (FddA). Representative suitable NNRTIs include nevirapine (BI-RG-587); delavirdine (BHAP, U-90152); efavirenz (DMP-266); PNU-142721; AG-1549; MKC-442 (1-(ethoxy-methyl)-5-(1-methylethyl)-6-(phenylmethyl)-(2,4(1H,3H)-pyrimidinedione); and (+)-calanolide A (NSC-675451) and B. Representative suitable protease inhibitors include saquinavir (Ro 31-8959); ritonavir (ABT-538); indinavir (MK-639); nelfinavir (AG-1343); amprenavir (141W94); lasinavir (BMS-234475); DMP-450; BMS-2322623; ABT-378; and AG-1549. Other antiviral drugs include hydroxyurea, ribavirin, IL-2, IL-12, pentafuside, and Yissum Project No. 11607.

[0185] Combination therapy is intended to include administration of the therapeutic agents in a sequential manner, i.e., each therapeutic agent is administered at various different times, as well as administration of the therapeutic agents or at least two therapeutic agents in a substantially simultaneous manner. Substantially simultaneous administration can be achieved, for example, by administering to the patient a single dosage form of each therapeutic agent with a fixed ratio, or a single dosage form of each therapeutic agent. Sequential or substantially simultaneous administration of each therapeutic agent can be performed by any suitable route, including, but not limited to, oral, intravenous, intramuscular, and direct absorption through mucosal membranes. The therapeutic agents can be administered by the same route or by different routes. For example, a first therapeutic agent in a selected combination can be administered by intravenous injection, while another therapeutic agent in the combination can be administered orally. Alternatively, for example, all therapeutic agents can be administered orally, or all therapeutic agents can be administered by intravenous injection. The administration of the therapeutic agents described above can also be combined with other biologically active ingredients and non-pharmaceutical therapies (e.g., surgery or radiation therapy) to perform combination therapy. When the combination therapy further includes a non-drug treatment, the non-drug treatment can be administered at any suitable time, so long as the beneficial effect resulting from the synergistic action of the combination of the therapeutic agent and the non-drug treatment is achieved, e.g., in suitable cases, the beneficial effect is achieved even when the non-drug treatment is temporarily suspended, perhaps for days or weeks, from administration of the therapeutic agent.

[0186] Pharmaceutical Composition The present invention also provides pharmaceutical compositions comprising a therapeutically effective amount of one or more compounds of formula (I), formulated together with one or more pharma- ceutically acceptable carriers (excipients) and / or diluents, and optionally one or more further therapeutic agents as described above.

[0187] The compounds of formula (I) may be administered by any suitable route, preferably in the form of a pharmaceutical composition adapted for such route, and in a dosage effective for the intended treatment. The compounds of formula (I) and compositions of compounds of formula (I) may be administered for any of the uses described herein by any suitable method, for example, oral administration (e.g., tablets, capsules (including sustained or timed release formulations, respectively), pills, powders, granules, elixirs, tinctures, suspensions (including nanosuspensions, microsuspensions, spray-dried dispersions), syrups, and emulsions); sublingual administration; buccal administration; parenteral administration (e.g., subcutaneous, intravenous, intramuscular, or intrasternal injection, or infusion techniques (e.g., sterile injectable aqueous or non-aqueous solutions or suspensions); nasal administration, including administration to the nasal membranes (e.g., inhalation sprays); topical administration (e.g., in the form of cream formulations or ointments); or rectal administration (e.g., in the form of suppositories). They may be administered alone, but will generally be administered with a pharmaceutical carrier selected on the basis of the selected route of administration and standard pharmaceutical practice.

[0188] For oral administration, the pharmaceutical composition may be in the form of, for example, a tablet, capsule, liquid capsule, suspension, or liquid. The pharmaceutical composition is preferably formulated in a dosage unit form with a specific amount of active ingredient. For example, the pharmaceutical composition may be provided as a tablet or capsule containing an amount of active ingredient ranging from about 0.1 to 1000 mg, preferably from about 0.25 to 250 mg, more preferably from about 0.5 to 100 mg. The appropriate daily dose to be administered to a human or other mammal may vary widely depending on the patient's condition and other factors, but can be determined using routine methods.

[0189] Any pharmaceutical composition discussed herein can be delivered orally, for example, via any acceptable and suitable oral formulation. Examples of oral formulations include, but are not limited to, tablets, troches, lozenges, aqueous and oily suspensions, dispersible powders or granules, emulsions, hard and soft capsules, liquid capsules, syrups, and elixirs. Pharmaceutical compositions for oral administration can be prepared according to any method known in the art for preparing pharmaceutical compositions for oral administration. In order to provide a medicament that is easy to swallow, the pharmaceutical compositions described in the present invention can include at least one substance selected from sweeteners, flavoring agents, coloring agents, demulcents, antioxidants, and preservatives.

[0190] Tablets can be prepared, for example, by mixing at least one compound of formula (I) and / or at least one pharma- ceutically acceptable salt thereof with at least one non-toxic, pharma-ceutically acceptable additive suitable for tablet preparation. Examples of additives include, but are not limited to, inert diluents (e.g., calcium carbonate, sodium carbonate, lactose, calcium phosphate, and sodium phosphate), granulating and disintegrating agents (e.g., microcrystalline cellulose, croscarmellose sodium, corn starch, and alginic acid), binding agents (e.g., starch, gelatin, polyvinylpyrrolidone, and gum arabic), and lubricants (e.g., magnesium stearate, stearic acid, and talc). In addition, tablets can be uncoated or coated by known techniques to mask the unpleasant taste of unpleasant drugs or to delay disintegration and absorption of the active ingredient in the digestive tract, thus prolonging the effect of the active ingredient for a longer period of time. Examples of water-soluble taste-masking materials include, but are not limited to, hydroxypropyl methylcellulose and hydroxypropyl cellulose. Examples of time delay materials include, but are not limited to, ethyl cellulose and cellulose acetate butyrate.

[0191] Hard gelatin capsules can be prepared, for example, by mixing at least one compound of formula (I) and / or at least one salt thereof with at least one inert solid diluent (e.g., calcium carbonate, calcium phosphate, and kaolin).

[0192] Soft gelatin capsules can be prepared, for example, by mixing at least one compound of formula (I) and / or at least one pharma- ceutically acceptable salt thereof with at least one water-soluble carrier (e.g., polyethylene glycol) and at least one oil medium (e.g., peanut oil, liquid paraffin, and olive oil).

[0193] Aqueous suspensions can be prepared, for example, by mixing at least one compound of formula (I) and / or at least one pharma- ceutically acceptable salt thereof with at least one excipient suitable for the preparation of aqueous suspensions. Examples of excipients suitable for the preparation of aqueous suspensions include, but are not limited to, suspending agents (e.g., sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, alginic acid, polyvinylpyrrolidone, gum tragacanth, and gum arabic), dispersing or wetting agents (e.g., naturally occurring phosphatides (e.g., lecithin), condensation products of alkylene oxides with fatty acids (e.g., polyoxyethylene stearate), condensation products of ethylene oxide with long chain aliphatic alcohols (e.g., heptadecaethyleneoxycetanol), condensation products of ethylene oxide with fatty acids and Examples of suitable condensation products include condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides (e.g., polyoxyethylene sorbitol monooleate), and condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides (e.g., polyethylene sorbitan monooleate). The aqueous suspension may also contain at least one preservative (e.g., ethyl p-hydroxybenzoate and n-propyl p-hydroxybenzoate), at least one coloring agent, at least one flavoring agent, and / or at least one sweetening agent (e.g., but not limited to, sucrose, saccharin, and aspartame).

[0194] Oil suspensions can be prepared, for example, by suspending at least one compound of formula (I) and / or at least one pharma- ceutically acceptable salt thereof in either vegetable oil (e.g., peanut oil, olive oil, sesame oil, and coconut oil) or mineral oil (e.g., liquid paraffin). Oil suspensions can also include at least one thickening agent (e.g., beeswax, hard paraffin, and cetyl alcohol). To provide a palatable oil suspension, at least one sweetener already described above and / or at least one flavoring agent can be added to the oil suspension. Oil suspensions can further include at least one preservative, including but not limited to, for example, an antioxidant (e.g., butylated hydroxyanisole, and alpha-tocopherol).

[0195] Dispersible powders and granules can be prepared, for example, by mixing at least one compound of formula (I) and / or at least one pharma- ceutically acceptable salt thereof with at least one dispersing agent and / or wetting agent, at least one suspending agent, and / or at least one preserving agent. Suitable dispersing agents, wetting agents, and suspending agents have already been described above. Examples of preservatives include, but are not limited to, antioxidants (e.g., ascorbic acid). In addition, dispersible powders and granules can also include at least one excipient (e.g., but are not limited to, sweeteners, flavoring agents, and coloring agents).

[0196] The emulsion of at least one compound of formula (I) and / or at least one pharma- ceutically acceptable salt thereof may be prepared, for example, as an oil-in-water emulsion. The oil phase of the emulsion containing the compound of formula (I) may be composed of known ingredients in a known manner. The oil phase may be provided, for example, but not limited to, vegetable oils (e.g., olive oil and peanut oil), mineral oils (e.g., liquid paraffin), and mixtures thereof. The oil phase may include only an emulsifier, but may also include a mixture of at least one emulsifier and a fat or oil, or both a fat and an oil. Suitable emulsifiers include, but are not limited to, naturally occurring phosphatides (e.g., soybean lecithin), esters or partial esters derived from fatty acids and hexitol anhydrides (e.g., sorbitan monooleate), and condensation products of partial esters and ethylene oxide (e.g., polyoxyethylene sorbitan monooleate). Preferably, a hydrophilic emulsifier is included together with a lipophilic emulsifier acting as a stabilizer. It is also preferred to include both oil and fat. Together, the emulsifier, with or without stabilizer, makes up the so-called emulsifying wax, and the wax, together with the oil and fat, makes up the so-called emulsifying ointment base, which forms the oily dispersed phase of the cream formulation. The emulsion may also include sweeteners, flavorings, preservatives, and / or antioxidants. Emulsifiers and emulsion stabilizers suitable for use in the formulation of the present invention include Tween 60, Span 80, cetostearyl alcohol, myristyl alcohol, glyceryl monostearate, sodium lauryl sulfate, glyceryl distearate, alone or with wax; or other materials known in the art.

[0197] The compound of formula (I) and / or at least one pharmaceutically acceptable salt thereof can be delivered, for example, intravenously, subcutaneously, and / or intramuscularly via any pharmaceutically acceptable and suitable injection form. Examples of injection forms include, but are not limited to, sterile aqueous solutions containing acceptable vehicles and solvents (e.g., water, Ringer's solution, and isotonic sodium chloride solution), sterile oil-in-water microemulsions, and aqueous or oily suspensions.

[0198] Preparations for parenteral administration may be in the form of aqueous or non-aqueous isotonic sterile injection solutions or suspensions. These solutions and suspensions may be prepared from sterile powders or granules by using one or more carriers or diluents described for use in preparations for oral administration, or by using other suitable dispersing or wetting agents and suspending agents. The compound may be dissolved in water, polyethylene glycol, propylene glycol, ethanol, corn oil, cottonseed oil, peanut oil, sesame oil, benzyl alcohol, sodium chloride solution, tragacanth gum, and / or various buffer solutions. Other adjuvants and methods of administration are known and widely known in the pharmaceutical field. The active ingredient may also be administered by injection in a composition with a suitable carrier (e.g., saline, dextrose, or water), or cyclodextrin (i.e., Captisol), solubilizing cosolvent (i.e., propylene glycol), or solubilizing micelle (i.e., Tween 80).

[0199] A sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent (e.g., a solution in 1,3-butanediol). Among the acceptable vehicles and solvents that may be used are water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile fixed oils are conventionally used as a solvent or suspending medium. For this purpose, any sterile fixed oil may be used, including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used as injectable preparations.

[0200] Sterile injectable oil-in-water microemulsions can be prepared, for example, by: 1) dissolving at least one compound of formula (I) in an oil phase (e.g., a mixture of soybean oil and lecithin), 2) combining the oil phase containing formula (I) with a mixture of water and glycerol, and 3) treating the combination to form a microemulsion.

[0201] Sterile aqueous suspension or sterile oily suspension can be prepared according to a method known to those skilled in the art. For example, sterile aqueous solution or sterile aqueous suspension can be prepared using a non-toxic, parenterally acceptable diluent or solvent (e.g., 1,3-butanediol), and sterile oily suspension can be prepared using a non-toxic, acceptable solvent or suspension medium (e.g., sterile fixed oil (e.g., synthetic monoglyceride or diglyceride), and fatty acid (e.g., oleic acid).

[0202] Pharmaceutically acceptable carriers are formulated according to many factors well within the expertise of those skilled in the art. These factors include, but are not limited to, the type and nature of the active agent to be formulated, the patient to whom the composition containing the active agent will be administered, the intended route of administration of the composition, and the therapeutic indications to be targeted. Pharmaceutically acceptable carriers include both aqueous and non-aqueous liquid media, as well as various solid and semi-solid dosage forms. Such carriers can include many different components and additives in addition to the active agent, and such additional components are included in the formulation for various reasons, such as stabilization of the active agent, binders, etc., as known to those skilled in the art. Descriptions of suitable pharmacopoeia acceptable carriers and factors in selecting them can be found in a variety of readily available literature, such as Allen, LV Jr. et al. Remington: The Science and Practice of Pharmacy (2 Volumes), 22nd Edition (2012), Pharmaceutical Press.

[0203] Pharmaceutically acceptable carriers, adjuvants, and vehicles that may be used in the pharmaceutical compositions of the present invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, self-emulsifying drug delivery systems (SEDDS) (e.g., d-α-tocopherol polyethylene glycol 1000 succinate), surfactants used in pharmaceutical dosage forms (e.g., Tween, polyethoxylated castor oil (e.g., CREMOPHOR surfactants (BASF), or other similar polymeric delivery matrices), serum proteins (e.g., human serum albumin), buffer substances (e.g., phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts, or electrolytes (e.g., protamine sulfate, glyceryl stearate, sorbitol ... Examples of suitable cyclodextrins include cyclodextrins such as α-, β-, and γ-cyclodextrins, or chemically modified derivatives such as hydroxyalkylcyclodextrins, including 2- and 3-hydroxypropylcyclodextrins, or other solubilizing derivatives.

[0204] The pharma- ceutical active compounds of the present invention can be processed according to conventional pharmaceutical methods to prepare medicaments for administration to patients (e.g., humans and other mammals). The pharmaceutical compositions can be subjected to conventional pharmaceutical operations (e.g., sterilization) and / or can contain conventional adjuvants (e.g., preservatives, stabilizers, wetting agents, emulsifiers, buffers, etc.). Tablets and pills can additionally be prepared with enteric coatings. Such compositions can also contain adjuvants (e.g., wetting agents, sweeteners, flavoring agents, and aromatic agents).

[0205] For treatment, the active compound of the present invention is usually combined with one or more adjuvants suitable for intended administration route.When administered orally, the compound can be mixed with lactose, sucrose, starch powder, cellulose ester of alkanoic acid, cellulose alkyl ester, talc, stearic acid, magnesium stearate, magnesium oxide, sodium and calcium salts of phosphoric acid and sulfate, gelatin, gum arabic, sodium alginate, polyvinylpyrrolidone, and / or polyvinyl alcohol, and then tableted or encapsulated for convenient administration.Such capsules or tablets can include controlled release formulations, and can be provided with active compound dispersed in hydroxypropylmethylcellulose.

[0206] The amount of compound administered to treat a condition using the compounds and / or compositions of the present invention, and the dosing schedule will depend on a variety of factors, such as age, weight, sex, the condition of the patient, the type of disease, the severity of the disease, the route and frequency of administration, and the specific compound utilized. Thus, the dosing schedule may vary widely, but can be routinely determined using standard methods. A daily dose of about 0.001 to 100 mg / kg body weight, preferably between about 0.0025 to about 50 mg / kg body weight, and most preferably between about 0.005 to 10 mg / kg body weight, may be appropriate. The daily dose may be administered 1 to 4 times a day. Other dosing schedules include weekly and biday cycles.

[0207] The pharmaceutical compositions of the present invention include at least one compound of formula (I) and / or at least one pharma- ceutically acceptable salt thereof, and optionally an excipient selected from any pharma- ceutically acceptable carrier, adjuvant, and vehicle. Another composition of the present invention includes a compound of formula (I) as described herein, or a prodrug thereof, and a pharma- ceutically acceptable carrier, adjuvant, or vehicle.

[0208] The present invention also includes pharmaceutical kits useful, for example, in the treatment or prevention of Helios protein-associated diseases or disorders, and other diseases described herein, which include one or more containers containing a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I). Such kits may further include one or more of a variety of conventional pharmaceutical kit components, as desired (e.g., a container containing one or more pharma- ceutically acceptable carriers, another container), which will be readily apparent to those of skill in the art. Instructions, either in the form of a package insert or label, indicating the amounts of components to be administered, administration guidelines, and / or mixing guidelines for the components, may also be included in the kit.

[0209] Dosing regimens for the compounds of the invention will, of course, vary depending on known factors, such as the pharmacodynamic properties of the particular agent and its method and route of administration; the species, age, sex, health, medical condition, and weight of the recipient; the nature and extent of the condition; type of concomitant treatment; frequency of treatment; route of administration, the patient's renal and hepatic function, and the desired effect.

[0210] As a general guidance, the daily oral dose of each active ingredient, when used to obtain the intended effect, will range from about 0.001 to about 5000 mg / day, preferably about 0.01 to about 1000 mg / day, and most preferably between about 0.1 and about 250 mg / day. The most preferred dose for intravenous constant rate infusion is in the range of about 0.01 to about 10 mg / kg / min. Compounds of formula (I) may be administered in a single daily dose or in divided doses of two, three, or four total daily doses.

[0211] The compounds will generally be administered in admixture with suitable pharmaceutical diluents, excipients, or carriers (collectively referred to herein as pharmaceutical carriers) appropriately selected for the intended form of administration (e.g., oral tablets, capsules, elixirs, and syrups) and consistent with conventional pharmaceutical standards.

[0212] Dosage forms (pharmaceutical compositions) suitable for administration may contain about 1 mg to about 200 mg of active ingredient per dosage unit. In these pharmaceutical compositions, the active ingredient is usually present in an amount of about 0.1 to 95% by weight of the total weight of the composition.

[0213] A typical capsule for oral administration contains at least one compound of formula (I) (250 mg), lactose (75 mg), and magnesium stearate (15 mg), which mixture is passed through a 60 mesh sieve and filled into a No. 1 gelatin capsule.

[0214] A typical injectable formulation is prepared by aseptically adding at least one compound of formula (I) (250 mg) to a vial, aseptically lyophilizing and sealing the vial, and at the time of use, mixing the contents of the vial with saline (2 mL) to prepare the injectable formulation.

[0215] The present invention includes within its scope pharmaceutical compositions comprising a therapeutically effective amount of at least one compound of formula (I) as an active ingredient, alone or in combination with a pharmaceutical carrier. Compounds of formula (I) may be used alone, in combination with other compounds of formula (I), or in combination with one or more other therapeutic agents (e.g., anti-cancer agents or other pharma- ceutical active substances), as appropriate.

[0216] Regardless of the route of administration selected, the compounds of formula (I) and / or pharmaceutical compositions of the present invention, which may be used in a suitable hydrated form, are formulated into pharma- ceutical acceptable dosage forms by conventional methods known to those of skill in the art.

[0217] Actual dosage levels of the active ingredient in the pharmaceutical compositions of formula (I) may be varied to contain an amount of the active ingredient effective to obtain a therapeutic effect for a particular patient, composition and mode of administration, without being toxic to the patient.

[0218] The selected dosage level will depend upon a variety of factors, including the activity of the particular compound of formula (I) or its esters, salts or amides employed, the route of administration of the particular compound employed, the time of administration, the rate of excretion or metabolism, the rate and extent of absorption, the duration of treatment, other drugs, compounds and / or substances used in combination with the particular compound employed, the age, sex, weight, symptoms, health and medical history of the patient being treated, and factors well known in the medical arts.

[0219] A physician or veterinarian having ordinary skill in the art can readily determine and prescribe the effective amount of the pharmaceutical composition required. For example, the physician or veterinarian can start the dosage of the compound of formula (I) used in the pharmaceutical composition at a lower level than required to achieve a therapeutic effect, and gradually increase the dosage until an effect is achieved.

[0220] In general, the suitable daily dose of the compound of formula (I) is the lowest effective dose of the compound to obtain a therapeutic effect. Such an effective dose is generally determined by the above factors. In general, the dose of the compound of formula (I) to a patient is about 0.01 to about 50 mg / kg body weight / day for oral, intravenous, intracerebroventricular and subcutaneous administration.

[0221] If desired, the effective daily dose of the active compound may be administered in two, three, four, five, six or more divided doses at appropriate intervals throughout the day, conveniently in unit dosage forms. In some embodiments of the invention, dosing is once a day.

[0222] While it is possible for a compound of formula (I) to be administered alone, it is preferable to administer the compound as a pharmaceutical formulation (composition).

[0223] The other therapeutic agents described above, when used in combination with the compounds of formula (I), may be used, for example, in amounts as set forth in the Pharmaceutical Description (PDR) or as determined by one of ordinary skill in the art. In the methods of the invention, the other therapeutic agents may be administered prior to, simultaneously with, or after administration of the compounds of the invention.

[0224] (Manufacturing method) The compounds of the present invention can be prepared by many methods known to those skilled in the art of organic synthesis. The compounds of the present invention can be synthesized by the following methods using synthetic methods known in the art of organic synthetic chemistry, or by analogous synthetic methods that will be appreciated by those skilled in the art. Preferred methods include, but are not limited to, the following methods. All documents cited herein are incorporated by reference in their entirety.

[0225] The compounds of the present invention may be prepared using the reactions and techniques described in this section. The reactions are carried out in solvents suitable for the reagents and materials used and are appropriate for the transformations effected. It is also understood that in the description of the synthetic methods described below, all of the reaction conditions presented (including the choice of solvent, reaction atmosphere, reaction temperature, experimental time, and work-up method) are selected to be conditions that are standard for the reactions, which should be readily recognized by those skilled in the art. It is understood by those skilled in the art of organic synthesis that the functional groups present on the various parts of the molecule must be compatible with the reagents and reactions presented. Such restrictions on the substituents compatible with the reaction conditions will be readily apparent to those skilled in the art, and alternative methods must be used. The reactions may require the decision to change the order of synthetic steps or to select certain reaction courses differently in order to obtain the desired compounds of the present invention. It is also recognized that another important consideration in planning any synthetic route in this field is the selection of appropriate protecting groups to be used to protect reactive functional groups present in the compounds described in the present invention. For the experienced experimenter, an authoritative reference describing many protecting group alternatives is Protective Groups In Organic Synthesis by Greene and Wuts (Fourth Edition, Wiley & Sons, 2007).

[0226] Compounds of formula (I) may be prepared by reference to the methods depicted in the following schemes. As depicted in the schemes, the final product is a compound having the same structural formula as formula (I). It is understood that by selecting appropriate reagents for the appropriate substituents, any compound of formula (I) may be prepared by the schemes. Solvents, temperatures, pressures and other reaction conditions may be readily selected by one of ordinary skill in the art. Starting materials are commercially available or readily prepared by one of ordinary skill in the art. The components of the compounds are defined as described herein.

[0227] The general synthetic route of the compounds according to the invention is illustrated in Schemes 1-2, where the substituents R1, R2, R4 and R6 are as defined above or are functional groups which may be converted to the final substituents. The substituent X is a leaving group, such as a halide (preferably I, Br or Cl) or a triflate. The substituent M is a suitable coupling partner, such as a boronic acid, a boronic ester or a stannane. The substituent R is a carboxylic acid protecting group, such as tert-butyl, methyl, ethyl or benzyl. As shown in Scheme 1, the general method for preparing the compounds of the invention can start with a suitable substituted aryl fluoride 1. Treatment with a suitable nucleophile, for example intermediate 2, where M can be MgBr or Li, can produce a substituted aryl halide such as 3. Halogenation, preferably bromination, can be achieved by treating 3 with a reagent such as N-bromosuccinimide to give bromide 4. Treatment of 4 with amine 5 under basic conditions such as potassium carbonate results in initial displacement of the secondary bromide, followed by subsequent cyclization to give lactam 6.

[0228] [ka]

[0229] When M is a stannane, 6 can be coupled to an appropriately substituted heterocycle 7 via a Stille coupling reaction using a suitable catalyst (e.g., Pd(PPh3)4 or bis(triphenylphosphine)dichloropalladium(II) / CuI) to provide 8. Alternatively, 6 can be converted to a boronic acid or boronic ester 9 using conditions well known to those skilled in the art. The boronic acid or boronic ester 9 can be reacted with a suitable palladium catalyst (e.g., Pd(PPh3)4 or 1,1'-bis(diphosphinylferino)ferrocene]dichloropalladium(II) in the presence of a suitable base (e.g., cesium carbonate, potassium phosphate, or sodium bicarbonate) to provide 8.

[0230] Depending on which acid protecting group R is selected in intermediate 8, different conditions are required for conversion to compound 11 (Scheme 2). For example, when R is methyl, ethyl or benzyl, base-induced cyclization of 8 is preferred to convert 8 directly to 11 using a suitable base (e.g. LiHMDS) in a suitable solvent (e.g. tetrahydrofuran). When R is tert-butyl, acid-induced cyclization of 12 is preferred to convert 8 directly to 11 using a suitable acid (e.g. benzenesulfonic acid) in a suitable solvent (e.g. acetonitrile). In other cases, a two-step method is preferred, first generating the corresponding free carboxylic acid 8 using conditions appropriate for the particular acid protecting group R. These are methods well known to those skilled in the art of organic synthesis. For example, when R is tert-butyl, acid hydrolysis is preferred using a suitable acid (e.g. trifluoroacetic acid or hydrochloric acid). When R is methyl, ethyl or benzyl, base hydrolysis is preferred using a suitable base (e.g. LiOH). In other cases, when R is benzyl, deprotection by palladium catalyzed hydrogenolysis is preferred. Once the carboxylic acid is generated, it can be activated by the action of thionyl chloride / dimethylformamide or carbonyldiimidazole / dimethylaminopyridine and subjected to intramolecular attack by the pendant primary amine to give 11.

[0231] [ka] EXAMPLES

[0232] (Example) The following examples are illustrative of certain embodiments of the invention and are not intended to limit the scope of the invention. Chemical abbreviations and symbols, as well as scientific abbreviations and symbols, have their usual and customary meanings unless otherwise specified. Additional abbreviations used in the examples and elsewhere in this specification are defined above. Common intermediates are generally useful in the preparation of one or more of the examples. Compounds of the examples are identified by the example and step by which they are prepared (e.g., "1-A" represents step A of Example 1), or by the example only if the compound is the title compound of the example (e.g., "1" represents the title compound of Example 1). In some cases, alternative methods of preparation of intermediates or examples are described. Chemists skilled in the synthetic arts will frequently devise alternative preparation methods that are desirable based on one or more considerations (e.g., shorter reaction times, cheaper starting materials, ease of handling and purification, higher yields, ease of catalysis, avoidance of toxic reagents, availability of specialized equipment, and fewer steps). The intent of describing alternative preparation methods is to make the examples of the invention more amenable to preparation. In some cases, some of the functional groups in the outlined examples and claims may be substituted with bioisosteric replacements known to those of skill in the art (e.g., replacing a carboxylic acid group with a tetrazole or phosphate moiety).

[0233] [Table 8]

[0234] Preparative HPLC Method 1: XBridge C18, 200mm x 19mm, particle size: 5μm; Mobile phase A: 5:95 acetonitrile:water (containing 10mM ammonium acetate); Mobile phase B: 95:5 acetonitrile:water (containing 10mM ammonium acetate); Gradient: 15% B hold for 0 min, 15-50% B over 25 min, then 100% B hold for 6 min; Flow rate: 20mL / min; Column temperature 25°C. Fraction collection was triggered by the MS signal.

[0235] Example 1 2-Amino-6-(2-(2,6-dioxopiperidin-3-yl)-4-methyl-1-oxoisoindolin-5-yl)-4-methylnicotinonitrile [ka] Preparation 1A: tert-butyl (S)-5-amino-4-(5-bromo-4-methyl-1-oxoisoindolin-2-yl)-5-oxopentanoate To a suspension of tert-butyl (S)-4,5-diamino-5-oxopentanoic acid hydrochloride (2.162 g, 6.06 mmol) in acetonitrile (30 ml) at 0° C. was added DIPEA (3.018 ml, 17.28 mmol). After stirring for 20 min, the reaction mixture was treated with a solution of methyl 4-bromo-2-(bromomethyl)-3-methylbenzoate (2.650 g, 8.22 mmol) in MeCN (10 mL). The reaction mixture was stirred at 0° C. for 5 min. The ice bath was removed and the reaction mixture was allowed to warm to room temperature and stirred at room temperature for 1 h. The reaction mixture was warmed to 60° C. and maintained at that temperature overnight. The reaction mixture was concentrated, dissolved in EtOAc, washed twice with water, then with 1.5M K2HPO4, then with brine, dried over MgSO4, filtered, and concentrated. This material was purified on silica gel eluting with 30-100% EtOAc / Hex The resulting material was triturated with ether (8 mL) to give the title product in 33.7% yield. 1H NMR(400 MHz, CHLOROFORM-d) δ 7.70(d, J=8.1 Hz, 1H), 7.56(d, J=8.1 Hz, 1H), 6.23(br s, 1H), 5.31(br s, 1H), 4.92(dd, J=8.6, 6.4 Hz, 1H), 4.55-4.47(m, 1H), 4.45-4.36(m, 1H), 2.41(s, 3H), 2.41-2.14(m, 4H), 1.45(s, 9H).

[0236] Preparation 1B: tert-Butyl (S)-5-amino-4-(4-methyl-1-oxo-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoindolin-2-yl)-5-oxopentanoate A dried flask was charged with tert-butyl (S)-5-amino-4-(5-bromo-4-methyl-1-oxoisoindolin-2-yl)-5-oxopentanoate (300 mg, 0. 730 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (278 mg, 1.094 mmol) and potassium acetate (214 mg, 2.188 mmol) and purged with nitrogen. The solid was suspended in dioxane (10 mL) and degassed by bubbling nitrogen through it with stirring for 5 minutes. The reaction mixture was treated with Pd(dppf)Cl2 (16.02 mg, 21.88 μmol). The flask was degassed for 5 minutes, sealed and heated to 60° C. under nitrogen for 18 hours. The reaction mixture was diluted with EtOAc, washed with brine, and dried over MgSO4. It was concentrated and purified on a 40 g silica gel column by ISCO, eluted with 0-40% B / DCM (where B=15% EtOH / EtOAc+0.1% TEA) to give the product in 99% yield. MS (ES): m / z = 459.3 [M+H] + .

[0237] Example 1: To a microwave vial (2 mL) was added 2-amino-6-bromo-4-methylnicotinonitrile (12 mg, 0.057mmol), tert-butyl (S)-5-amino-4-(4-methyl-1-oxo-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoindolin-2-yl)-5-oxopentanoate (28.5 mg, 0.062 mmol), Pd(dtbpf)Cl2 (1.106 mg, 1.698 μmol), dioxane (1.5 mL) and aqueous K3PO4 (0.075 mL, 0.226 mmol). The vial was sealed and the air was replaced with nitrogen. The reaction mixture was heated in a microwave at 120 °C for 15 min. After cooling to room temperature, the reaction mixture was diluted with EtOAc, washed with brine and the organic layer was separated and concentrated. The resulting residue was dissolved in 1 mL of PhSO3H solution (1.44 g / 40 mL) / MeCN and heated in a microwave at 120° C. for 10 min. The material was concentrated to dryness and the residue was dissolved in DMSO (1.9 mL) and purified by preparative HPLC method 1 to give the title compound in 49% yield. MS (ES): m / z = 390.3 [M+H] + . 1 H NMR(500 MHz, DMSO-d6) δ ppm 11.03(s, 1 H) 7.66(d, J=7.63 Hz, 1 H) 7.53(d, J=7.93 Hz, 1 H) 6.91(s, 2 H) 6.78(s, 1 H) 5.18(br dd, J=13.43, 5.19 Hz, 1 H) 4.52(br d, J=17.09 Hz, 1 H) 4.35(br d, J=17.40 Hz, 1 H) 2.91-3.01(m, 1 H) 2.65(br d, J=16.78 Hz, 1 H) 2.47(br dd, J=13.12, 4.27 Hz, 1 H) 2.43(s, 3 H) 2.33(s, 3 H) 2.03-2.10(m, 1 H).

[0238] Example 2 2-Amino-6-(2-(2,6-dioxopiperidin-3-yl)-6-methyl-1-oxoisoindolin-5-yl)-4-methylnicotinonitrile [ka] Preparation 2A: Methyl 4-bromo-2-(bromomethyl)-5-methylbenzoate To a solution of 4-bromo-2,5-dimethylbenzoic acid (5.0 g, 21.82 mmol) at room temperature, SOCl2 (31.66 mL, 436 mmol) was added and stirred for 2 h. The formation of the acid chloride was confirmed by LCMS, and then the mixture was concentrated to dryness. Methanol (30 mL) was added at room temperature, and the mixture was stirred for 0.5 h, and then concentrated to dryness again. To the resulting methyl ester, CCl4 (180 mL) was added, followed by 1-bromopyrrolidine-2,5-dione (4.08 g, 22.92 mmol), and then AIBN (0.108 g, 0.654 mmol). The resulting mixture was heated to 80° C. with stirring overnight. After cooling to room temperature, the mixture was concentrated, dissolved in EtOAc, washed with brine, and dried over MgSO4. Two major peaks were observed by LCMS. The resulting mixture of regioisomers (5.4 g) was used in the next step.

[0239] Preparation 2B: tert-butyl (S)-5-amino-4-(5-bromo-6-methyl-1-oxoisoindolin-2-yl)-5-oxopentanoate To a suspension of methyl 4-bromo-2-(bromomethyl)-5-methylbenzoate (5.4 g, 16.76 mmol) in acetonitrile (100 mL) was added tert-butyl (S)-4,5-diamino-5-oxopentanoate hydrochloride (4.00 g, 16.76 mmol) followed by Hunig's base (5.84 mL, 33.52 mmol). After stirring at room temperature for 1 h, the reaction mixture was placed in a 40 °C bath and stirred at that temperature for 6 days. The reaction mixture was cooled to room temperature, diluted with EtOAc, washed with brine, dried over MgSO4, filtered and concentrated. The residue was purified by ISCO [120 g column] eluting with 0-100% EtOAc / DCM to give the desired compound (2.80 g). Additionally, two additional by-products (methyl (S)-5-(((1-amino-5-(tert-butoxy)-1,5-dioxopentan-2-yl)amino)methyl)-4-bromo-2-methylbenzoate and tert-butyl (S)-5-amino-4-(((2-((R)-1-amino-5-(tert-butoxy)-1,5-dioxopentan-2-yl)-6-bromo-3-oxoisoindolin-5-yl)methyl)amino)-5-oxopentanoate) were isolated. MS(ES): m / z = 411.0 [M+H] - . 1 H NMR (400 MHz, Chloroform-d) δ ppm 7.70 (d, J=6.44 Hz, 2 H) 6.31 (br s, 1 H) 5.36 (br s, 1 H) 4.90 (dd, J=8.68, 6.34 Hz, 1 H) 4.51 (d, J=16.98 Hz, 1 H) 4.41 (d, J=16.98 Hz, 1 H) 2.51 (s, 3 H) 2.21-2.43 (m, 3 H) 2.12-2.19 (m, 1 H) 1.44 (s, 9 H).

[0240] Preparation 2C: tert-Butyl (S)-5-amino-4-(6-methyl-1-oxo-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoindolin-2-yl)-5-oxopentanoate A 40 mL pressure-resistant vial was charged with tert-butyl (S)-5-amino-4-(5-bromo-6-methyl-1-oxoisoindolin-2-yl)-5-oxopentanoate (600 mg, 1.458 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (556 mg, 2.188 mmol), and potassium acetate (430 mg, 4.376 mmol) and flushed with nitrogen. The solid was suspended in dioxane (20 mL) and degassed with a nitrogen stream for 5 min while stirring. The reaction mixture was treated with Pd(dppf)Cl2 (32.02 mg, 0.044 mmol) and degassed for 5 min. The vial was sealed and heated to 95 °C under nitrogen for 3 h. The reaction mixture was cooled to room temperature, diluted with EtOAc, washed with brine, and dried over MgSO4. The filtrate was concentrated and purified on a 40 g silica gel column by ISCO, eluted with (0%-40% B / DCM, where B=15% EtOH / EtOAc+0.1% TEA) to give the title compound in 95% yield. MS(ES): m / z = 459.3 [M+H] + .

[0241] Example 2: To a microwave vial (2 mL) was added 2-amino-6-bromo-4-methylnicotinonitrile (12 mg, 0.057 mmol), tert-butyl (S)-5-amino-4-(6-methyl-1-oxo-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoindolin-2-yl)-5-oxopentanoate (28.5 mg, 0.062 mmol), Pd(dtbpf)Cl2 (1.106 mg, 1.698 μmol), dioxane (1.5 mL) and aqueous K3PO4 (0.075 mL, 0.226 mmol). The vial was sealed and the air was replaced with nitrogen. The reaction mixture was heated in a microwave at 120 °C for 15 min. After cooling to room temperature, the reaction mixture was diluted with EtOAc, washed with brine, and the organic layer was separated and concentrated. The resulting residue was dissolved in 1 mL of PhSO3H / MeCN solution (1.44 g / 40 mL) and heated in microwave at 120 °C for 10 min. The mixture was concentrated to dryness and the residue was dissolved in DMSO (1.7 mL) and purified by preparative HPLC method 1 to give the title compound in 35% yield. MS (ES): m / z = 390.3 [M+H] + . 1 H NMR (500 MHz, DMSO-d6) δ ppm 10.97-11.02 (m, 1 H) 7.65 (s, 1 H) 7.55 (s, 1 H) 6.76 (s, 1 H) 5.13 (dd, J=13.31, 5.18 Hz, 1 H) 4.39-4.51 (m, 1 H) 4.28-4.39 (m, 1 H) 2.87-2.98 (m, 1 H) 2.58-2.65 (m, 1 H) 2.51 (d, J=1.74 Hz, 3 H) 2.42-2.46 (m, 1 H) 2.41 (br s, 3 H) 2.39-2.40 (m, 1 H) 1.99-2.06 (m, 1H)

[0242] Example 3 2-Amino-6-(2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxoisoindolin-5-yl)-4-methylnicotinonitrile [ka] Intermediate 3A: 5-Bromo-4-fluoro-3-hydroxyisobenzofuran-1(3H)-one To a stirred solution of 2,2,6,6-tetramethylpiperidine (7.07 mL, 41.6 mmol) in THF (150 mL) was added 2.5 M n-BuLi solution in hexane (16 mL, 40.0 mmol) at 0° C. The reaction mixture was stirred at 0° C. for 30 min. To this, a solution of 4-bromo-3-fluorobenzoic acid (3.5 g, 15.98 mmol) in anhydrous THF (100 mL) was added dropwise at −50° C. The reaction mixture was stirred at the same temperature for 3 h. Anhydrous DMF (2.48 mL, 32.0 mmol) was added at −50° C. and the reaction mixture was brought to room temperature and stirred for 16 h. The reaction mixture was quenched with 1.5 N HCl (100 mL). The reaction mixture was extracted with ethyl acetate (3×30 mL). The combined organic layers were washed with brine, dried over anhydrous Na2SO4, filtered and evaporated under reduced pressure. The residue was purified by flash chromatography (SiO2, 120 g column, 0-50% EtOAc / petroleum-ether) to give 5-bromo-4-fluoro-3-hydroxyisobenzofuran-1(3H)-one (1.0 g, 23% yield) as a yellow solid. LCMS (Method A): Retention time 0.48 min, [M+H] + 245.1, 247.1; 1 H NMR (400 MHz, acetonitrile-d3) δ 7.93 (dd, J = 8.0, 5.5 Hz, 1H), 7.59 (d, J = 8.0 Hz, 1H), 6.74 (br s, 1H), 5.94 (br s, 1H).

[0243] Intermediate 3B: tert-butyl (S)-5-amino-4-(5-bromo-4-fluoro-1-oxoisoindolin-2-yl)-5-oxopentanoate To a stirred solution of 5-bromo-4-fluoro-3-hydroxyisobenzofuran-1(3H)-one (1.7 g, 6.88 mmol) and tert-butyl (S)-4,5-diamino-5-oxopentanoate hydrochloride (1.67 g, 8.26 mmol) in DMF (30 mL) was added sodium triacetoxyborohydride (3.65 g, 17.21 mmol) at 0° C. The reaction mixture was allowed to warm to room temperature and stirred for 48 hours. The reaction mixture was diluted with ice water (50 mL) and the resulting white solid was filtered and dried under vacuum to give tert-butyl (S)-5-amino-4-(5-bromo-4-fluoro-1-oxoisoindolin-2-yl)-5-oxopentanoate (1.6 g, 50% yield) as a white solid. LCMS (Method A): Retention time 1.39 min, [M+H] + 413.9, 415.3; 1 H NMR (400 MHz, DMSO-d6) δ 7.85 (dd, J = 8.0, 6.0 Hz, 1H), 7.59 (br s, 1H), 7.51 (d, J = 8.0 Hz, 1H), 7.23 (br s, 1H), 4.77-4.59 (m, 3H), 2.26-2.13 (m, 3H), 2.08-1.96 (m, 1H), 1.34 (s, 9H).

[0244] Intermediate 3C: tert-butyl (S)-5-amino-4-(5-(6-amino-5-cyano-4-methylpyridin-2-yl)-4-fluoro-1-oxoisoindolin-2-yl)-5-oxopentanoate A stirred solution of 2-amino-6-chloro-4-methylnicotinonitrile (50 mg, 0.30 mmol) and hexamethylditin (0.093 mL, 0.45 mmol) in toluene (2 mL) was purged with argon for 5 minutes, followed by the addition of [1,1'-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) (9.72 mg, 0.015 mmol). The reaction mixture was stirred at 100°C for 2 hours, cooled to room temperature, and filtered. The filtrate was concentrated under reduced pressure to give the crude product. [M+H]+ 298.2. To a solution of the crude product in dioxane (2 mL) was added tert-butyl (S)-5-amino-4-(5-bromo-4-fluoro-1-oxoisoindolin-2-yl)-5-oxopentanoate (98 mg, 0.24 mmol). The reaction mixture was purged with argon for 5 min, bis(triphenylphosphine)palladium(II) chloride (16.60 mg, 0.024 mmol) was added, and the reaction mixture was heated at 100° C. for 16 h. The reaction mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure to give the crude product, which was purified by flash chromatography (SiO2, 80 g column, 0-100% B (B = 15% EtOH / EtOAc, 0.5% TEA) / chloroform) to give tert-butyl (S)-5-amino-4-(5-(6-amino-5-cyano-4-methylpyridin-2-yl)-4-fluoro-1-oxoisoindolin-2-yl)-5-oxopentanoate (60 mg, 40.7% yield). LCMS (Method A): Retention time 1.35 min, [M+H] + 468.3.

[0245] Example 3: To a stirred solution of intermediate 3C (60 mg, 0.13 mmol) in acetic acid (2 mL) was added benzenesulfonic acid (20.30 mg, 0.13 mmol). The reaction mixture was heated to 150 °C under microwave irradiation for 10 min. The volatiles were removed under reduced pressure and the resulting crude product was purified by preparative LC-MS (column: mobile phase A: 0.1% trifluoroacetic acid in water; mobile phase B: acetonitrile; gradient: 10-40% B over 20 min, then hold at 100% B for 5 min; flow rate: 15 mL / min). The desired fraction was lyophilized to give 2-amino-6-(2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxoisoindolin-5-yl)-4-methylnicotinonitrile (15 mg, 29% yield). LCMS (Method G): Retention time 1.62 min, [M+H] + 394.2; 1 H NMR (400 MHz, DMSO-d6) δ 11.01 (br s, 1H), 8.01 (t, J=7.3 Hz, 1H), 7.70 (d, J=7.9 Hz, 1H), 7.06 (d, J=1.5 Hz, 1H), 6.99 (s, 2H), 5.14 (dd, J=13.3, 5.1 Hz, 1H), 4.74-4.38 (m, 2H), 3.05-2.85 (m, 1H), 2.75-2.57 (m, 2H), 2.46 (s, 3H), 2.13-1.95 (m, 1H).

[0246] General method 2: A stirred solution of intermediate 3B (1 eq.) and arylstannane reagent (1 eq.) in dioxane (5 mL / mmol) was purged with argon for 5 min. Bis(triphenylphosphine)palladium(II) chloride (0.1 eq.) was added and the reaction mixture was heated at 100° C. for 16 h. The reaction mixture was cooled to room temperature, filtered through a pad of Celite, and the filtrate was concentrated under reduced pressure. The crude material was transferred to a microwave vial, PhSO3H (2 eq.) and acetic acid (5 mL / mmol) were added, and the mixture was heated at 150° C. for 10 min in a microwave reactor. The mixture was concentrated and the residue was purified by prep-HPLC.

[0247] Example 4 3-(5-(6-amino-4-methylpyridin-2-yl)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione [ka] Intermediate 4A: 4-Methyl-6-(trimethylstannyl)pyridin-2-amine A stirred solution of 6-bromo-4-methylpyridin-2-amine (75 mg, 0.382 mmol) and hexamethylditin (0.119 mL, 0.57 mmol) in toluene (5 mL) was purged with argon for 5 min, then [1,1'-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) (12.4 mg, 0.02 mmol) was added. The reaction mixture was stirred at 100° C. for 2 h, cooled to room temperature, and filtered. The filtrate was concentrated under reduced pressure to give 4-methyl-6-(trimethylstannyl)pyridin-2-amine (130 mg, 89% yield). LCMS (Method A): Retention time 1.62 min, [M+H] + 273.1.

[0248] Example 4: Stille coupling and cyclization were carried out according to general procedure 2 using Preparation 4A and Preparation 3B. MS (ES): m / z = [M+H] + 369.1; 1H NMR (400 MHz, DMSO-d6) δ = 11.02 (s, 1H), 8.02 (t, J = 7.2 Hz, 1H), 7.65 (d, J = 7.8 Hz, 1H), 6.84 (s, 1H), 6.34 (s, 1H), 6.03 (s, 2H), 5.15 (dd, J = 5.0, 13.3 Hz, 1H), 4.66-4.57 (m, 1H), 4.49-4.38 (m, 1H), 3.02-2.85 (m, 1H), 2.62 (br d, J = 17.4 Hz, 1H), 2.49-2.40 (m, 1H), 2.22 (s, 3H), 2.07-1.98 (m, 1H).

[0249] Example 5 3-(5-(6-amino-4-(trifluoroethyl)pyridin-2-yl)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione [ka] Intermediate 5A: A stirred solution of 6-chloro-4-(trifluoromethyl)pyridin-2-amine (75 mg, 0.382 mmol) and hexamethylditin (0.119 mL, 0.57 mmol) in toluene (5 mL) was purged with argon for 5 minutes before adding [1,1'-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) (12.4 mg, 0.02 mmol). The reaction mixture was stirred at 100°C for 2 hours, cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure to give 4-(trifluoromethyl)-6-(trimethylstannyl)pyridin-2-amine (130 mg, 89% yield). LCMS (Method A): Retention time 1.92 min, [M+H] + 327.1.

[0250] Example 5: Stille coupling and cyclization were carried out according to general procedure 1 using intermediate 5A and intermediate 3B. MS (ES): m / z = [M+H]+ 423.3; 1 H NMR (400 MHz, DMSO-d6) δ = 11.02 (s, 1H), 8.05 (t, J = 7.3 Hz, 1H), 7.69 (d, J = 7.8 Hz, 1H), 7.15 (s, 1H), 6.78 (d, J = 11.8 Hz, 3H), 5.15 (dd, J = 5.0, 13.3 Hz, 1H), 4.68-4.57 (m, 1H), 4.52-4.39 (m, 1H), 2.99-2.82 (m, 1H), 2.67-2.58 (m, 1H), 2.48-2.39 (m, 1H), 2.12-1.93 (m, 1H).

[0251] Example 6 3-(4-fluoro-5-(4-methyl-6-(methylamino)pyridin-2-yl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione [ka] Intermediate 6A: N,4-Dimethyl-6-(trimethylstannyl)pyridin-2-amine A stirred solution of 6-chloro-N,4-dimethylpyridin-2-amine (100 mg, 0.64 mmol) and hexamethylditin (0.199 mL, 0.96 mmol) in toluene (10 mL) was purged with argon for 5 min, then [1,1'-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) (20.8 mg, 0.032 mmol) was added. The reaction mixture was stirred at 100° C. for 2 h, cooled to room temperature, and filtered. The filtrate was concentrated under reduced pressure to give N,4-dimethyl-6-(trimethylstannyl)pyridin-2-amine (250 mg, 55.0% yield). LCMS (Method A): Retention time 1.29 min, [M+H] + 287.1.

[0252] Example 6: Stille coupling and cyclization were carried out according to general procedure 2 using intermediate 6A and intermediate 3B. MS (ES): m / z = [M+H] + 383.1; 1 H NMR (400 MHz, DMSO-d6) δ 11.03 (s, 1H), 8.02 (s, 1H), 7.72 (d, J = 7.5 Hz, 1H), 6.93 (s, 1H), 6.58 (br s, 1H), 5.16 (dd, J = 13.1, 5.0 Hz, 1H), 4.82-4.40 (m, 3H), 2.89 (s, 3H), 2.75-2.63 (m, 1H), 2.64-2.57 (m, 1H), 2.46-2.41 (m, 1H), 2.36-2.28 (m, 3H), 2.06-2.00 (m, 1H). 19 / 19.

[0253] Example 7 3-(5-(6-amino-4,5-dimethylpyridin-2-yl)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione [ka] Intermediate 7A: 3,4-Dimethyl-6-(trimethylstannyl)pyridin-2-amine A stirred solution of 6-chloro-3,4-dimethylpyridin-2-amine (75 mg, 0.48 mmol) and hexamethylditin (0.149 mL, 0.72 mmol) in toluene (3 mL) was purged with argon for 5 minutes before adding [1,1'-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) (15.6 mg, 0.024 mmol). The reaction mixture was stirred at 100°C for 2 hours, cooled to room temperature, and filtered. The filtrate was concentrated under reduced pressure to give 3,4-dimethyl-6-(trimethylstannyl)pyridin-2-amine (150 mg, 55.0% yield). LCMS (Method A): Retention time 1.16 min, [M+H]+ 287.2.

[0254] Example 7: Stille coupling and cyclization were carried out according to general procedure 1 using intermediate 7A and intermediate 3B. MS (ES): m / z = [M+H] + 383.0; 1 H NMR (400 MHz, DMSO-d6) δ 11.01 (br s, 1H), 8.05 (t, J = 7.3 Hz, 1H), 7.65 (d, J = 7.9 Hz, 1H), 6.93 (d, J = 1.5 Hz, 1H), 5.81 (s, 2H), 5.14 (dd, J = 13.3, 5.1 Hz, 1H), 4.71-4.34 (m, 2H), 3.01-2.88 (m, 1H), 2.67-2.58 (m, 2H), 2.46-2.41 (m, 1H), 2.23 (s, 3H), 2.04 (s, 3H).

[0255] Examples 8 and 9 3-((S)-5-(6-amino-4,5-dimethylpyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione and 3-((R)-5-(6-amino-4,5-dimethylpyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione [ka]

[0256] Intermediate 8A: 4-Bromo-2-ethylbenzoic acid To a solution of 4-bromo-2-fluorobenzoic acid (5 g, 22.83 mmol) in anhydrous THF (100 mL) was added a 1M solution of ethylmagnesium bromide (22.83 mL, 68.5 mmol) in THF at -78 °C for 15 min. The reaction mixture was slowly warmed to room temperature and stirred under nitrogen atmosphere for 12 h. The reaction mixture was quenched dropwise with MeOH (15 mL). The reaction mixture was concentrated under reduced pressure. The resulting residue was partitioned between EtOAc and 2M aqueous hydrochloric acid. The layers were separated and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with brine, dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give the crude product. The crude product was purified by flash chromatography (SiO2, 80 g column, 0-30% EtOAc / petroleum-ether) to give 4-bromo-2-ethylbenzoic acid (4 g, 76% yield). LCMS (Method D): Retention time 2.49 min, [M+H] + 228.8, 230.0; 1 H NMR (400 MHz, DMSO-d6) δ 13.04 (br s, 1H), 7.71 (d, J = 8.5 Hz, 1H), 7.55 (d, J = 2.0 Hz, 1H), 7.49 (dd, J = 8.0, 2.0 Hz, 1H), 2.92 (q, J = 7.2 Hz, 2H), 1.15 (t, J = 7.5 Hz, 3H).

[0257] Intermediate 8B: Methyl 4-bromo-2-ethylbenzoate To a stirred mixture of 4-bromo-2-ethylbenzoic acid (4.0 g, 17.46 mmol) and Cs2CO3 (11.38 g, 34.9 mmol) in DMF (40 mL) was added methyl iodide (2.18 mL, 34.9 mmol). The reaction mixture was stirred at room temperature for 14 h, filtered through a pad of Celite, and concentrated in vacuo. The residue was purified by flash chromatography (SiO2, 80 g column, 0~50% EtOAc / petroleum ether) to give methyl 4-bromo-2-ethylbenzoate (3.3 g, 78% yield) as a colorless oil. 1H NMR (400 MHz, chloroform-d) δ 7.76 (d, J = 8.5 Hz, 1H), 7.46 (d, J = 1.5 Hz, 1H), 7.40 (dd, J = 8.5, 2.0 Hz, 1H), 3.91 (s, 3H), 2.99 (q, J = 7.5 Hz, 2H), 1.26 (t, J = 7.5 Hz, 3H).

[0258] Intermediate 8C: Methyl 4-bromo-2-(1-bromoethyl)benzoate To a stirred solution of methyl 4-bromo-2-ethylbenzoate (3.0 g, 12.34 mmol) in benzene (40 mL) was added NBS (3.29 g, 18.51 mmol) followed by AIBN (0.405 g, 2.47 mmol). The reaction mixture was heated at 85° C. for 15 h. The reaction mixture was cooled to room temperature, diluted with EtOAc, and washed with saturated aqueous 10% sodium thiosulfate and brine. The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (SiO2, 40 g column, 0-30% EtOAc / petroleum ether) to give methyl 4-bromo-2-(1-bromoethyl)benzoate (2.1 g, 53% yield) as a white solid. 1 H NMR (400 MHz, chloroform-d) δ 7.96 (d, J = 2.0 Hz, 1H), 7.73 (s, 1H), 7.49 (dd, J = 8.5, 2.0 Hz, 1H), 6.28 (q, J = 7.0 Hz, 1H), 3.95 (s, 3H).

[0259] Intermediate 8D: tert-Butyl (4S)-5-amino-4-(5-bromo-3-methyl-1-oxoisoindolin-2-yl)-5-oxopentanoate To a stirred solution of methyl 4-bromo-2-(1-bromoethyl)benzoate (1.0 g, 3.11 mmol) and H-Glu(OtBu)-NH2HCl (0.754 g, 3.73 mmol) in acetonitrile (30 mL) was added DIPEA (2.71 mL, 15.53 mmol). The resulting reaction mixture was heated at 85 °C for 15 h. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by flash chromatography (SiO2, 40 g column, 0-10% MeOH / DCM) to give tert-butyl (4S)-5-amino-4-(5-bromo-3-methyl-1-oxoisoindolin-2-yl)-5-oxopentanoate (650 mg, 51% yield) as a semi-solid. LCMS (Method A): Retention time 1.45 min, [M+H] + 411.3, 413.3.

[0260] Intermediate 8E: tert-butyl 5-amino-4-(5-(6-amino-4,5-dimethylpyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)-5-oxopentanoate A stirred solution of 6-chloro-3,4-dimethylpyridin-2-amine (200 mg, 1.28 mmol) and hexamethylditin (544 mg, 1.66 mmol) in toluene (6 mL) was purged with argon for 5 minutes, followed by the addition of [1,1'-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) (83 mg, 0.13 mmol). The reaction mixture was stirred at 100° C. for 2 hours, cooled to room temperature, and filtered. The filtrate was concentrated under reduced pressure to give the crude product. [M+H]+ 287.0. To a solution of this crude product in dioxane (4 mL) was added tert-butyl 5-amino-4-(5-bromo-3-methyl-1-oxoisoindolin-2-yl)-5-oxopentanoate (0.289 g, 0.70 mmol). The reaction mixture was purged with argon for 5 minutes. Bis(1,2-bis(diphenylphosphino)ethane)palladium(0) (0.063 g, 0.07 mmol) was added and the reaction mixture was heated at 100° C. for 16 h. The reaction mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure to give the crude product, which was purified by flash chromatography (SiO2, 80 g column, 0~100% B (B = 15% EtOH / EtOAc + 0.5% TEA) / chloroform) to give tert-butyl 5-amino-4-(5-(6-amino-4,5-dimethylpyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)-5-oxopentanoate (110 mg, 34.6% yield). LCMS (Method A): Retention time 1.40 min, [M+H] + 453.3.

[0261] Intermediate 8F: 3-(5-(6-amino-4,5-dimethylpyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione To a stirred solution of tert-butyl 5-amino-4-(5-(6-amino-4,5-dimethylpyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)-5-oxopentanoate (110 mg, 0.243 mmol, 34.6% yield) in acetonitrile (5 mL) was added p-toluenesulfonic acid (84 mg, 0.442 mmol). The reaction mixture was heated to 120° C. under microwave irradiation for 30 min. Volatiles were removed under reduced pressure and the crude product was purified by preparative HPLC (Column: Hypersil gold c18 (19 x 250 mm), 5 μm; Mobile phase A: 10 mM ammonium acetate / water, Mobile phase B: ACN; Flow rate: 20 mL T / B%: 0 / 20, 18 / 80, 19 / 100, 21 / 100, 22 / 20, 24 / 20). The desired fractions were lyophilized to give 3-(5-(6-amino-4,5-dimethylpyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione. LCMS (Method D): Retention time 2.15 min, [M+H] + 379.0.

[0262] Examples 8 and 9: 3-(5-(6-amino-4,5-dimethylpyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (100 mg) was subjected to SFC separation (column: Welk-01(R,R)(250×4.6) mm, 5 μm; % CO2: 45%; % co-solvent: 5 mM ammonium acetate / methanol and acetonitrile (1:1); flow rate: 4 g / min; temperature: 30° C.; UV: 237 nm), and the fraction of the first peak eluted at a retention time of 2.64 min was concentrated and lyophilized to give 3-((S)-5-(6-amino-4,5-dimethylpyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (12 mg, 15% yield). LCMS (Method D): Retention time 2.112 min, [M+H] + 379.0; 1H NMR (400 MHz, DMSO-d6) δ 11.10-10.71 (s, 1H), 8.19 (s, 1H), 8.12 (d, J=7.9 Hz, 1H), 7.69 (d, J=8.0 Hz, 1H), 7.14 (s, 1H), 5.73 (s, 2H), 4.93-4.67 (m, 2H), 2.85-2.71 (m, 2H), 2.25 (s, 3H), 2.04 (s, 3H), 1.51 (d, J=6.6 Hz, 3H) and the second peak fraction eluting at retention time 4.03 min were concentrated and lyophilized to give 3-((R)-5-(6-amino-4,5-dimethylpyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (8 mg, 10% yield). LCMS (Method D): Retention time 2.162 min, [M+H] + 379.0; 1 H NMR (400 MHz, DMSO-d6) δ 11.00-10.89 (s, 1H), 8.18 (s, 1H), 8.13 (d, J=8.0 Hz, 1H), 7.70 (d, J=8.0 Hz, 1H), 7.13 (s, 1H), 5.73 (s, 2H), 4.84-4.68 (m, 2H), 2.91-2.65 (m, 3H), 2.25 (s, 3H), 2.04 (s, 3H), 1.51-1.46 (m, 3H).

[0263] Examples 10 and 11 2-Amino-6-((3S)-2-(2,6-dioxopiperidin-3-yl)-3-methyl-1-oxoisoindolin-5-yl)-4-methylnicotinonitrile and 2-Amino-6-((3R)-2-(2,6-dioxopiperidin-3-yl)-3-methyl-1-oxoisoindolin-5-yl)-4-methylnicotinonitrile [ka] Intermediates 10A and 11A: 2-Amino-6-chloro-4-methylnicotinonitrile (300 mg, 1.790 mmol) and 1,1,1,2,2,2-hexamethyldistannane (762 mg, 2.327 mmol) in toluene (10 mL) were purged with argon for 5 minutes, followed by the addition of [1,1'-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) (117 mg, 0.179 mmol). The reaction mixture was stirred at 100° C. for 2 hours, cooled to room temperature, and filtered. The filtrate was concentrated under reduced pressure to give the crude product. [M+H]+ 298.0. The crude product was dissolved in dioxane and Intermediate 8D (0.695 g, 1.689 mmol) was added. The reaction mixture was purged with argon for 5 min and bis(diphenylphosphino)ethane)palladium(0) (0.153 g, 0.169 mmol) was added. The reaction mixture was heated to 100° C. and stirred for 16 h. The reaction mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure to give the crude product, which was purified by flash chromatography (SiO2, 80 g column, 0-100% B (B=15% EtOH in EtOAc, 0.5% TEA) / chloroform) to give tert-butyl 5-amino-4-(5-(6-amino-5-cyano-4-methylpyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)-5-oxopentanoate (200 mg, 0.431 mmol, 25.5% yield). LCMS (Method A): Retention time 1.47 min, [M+H] + 464.3.

[0264] Examples 10 and 11: To a stirred solution of tert-butyl 5-amino-4-(5-(6-amino-5-cyano-4-methylpyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)-5-oxopentanoate (200 mg, 0.431 mmol) in acetonitrile (10 mL) was added p-toluenesulfonic acid (164 mg, 0.863 mmol). The reaction mixture was heated to 120° C. under microwave irradiation for 30 min. Volatiles were removed under reduced pressure and the resulting crude product was purified by preparative HPLC (Column: Hypersil gold c18 (19 x 250 mm), 5 μm Mobile phase A-10 mM ammonium acetate / water; Mobile phase B: ACN; Flow rate: 20 mL T / B%: 0 / 20, 18 / 80, 19 / 100, 21 / 100, 22 / 20, 24 / 20). The desired fraction was lyophilized to give 2-amino-6-(2-(2,6-dioxopiperidin-3-yl)-3-methyl-1-oxoisoindolin-5-yl)-4-methylnicotinonitrile as a mixture of diastereomers. LCMS (Method D): Retention time 1.66 min, [M+H]+ 390.2. The diastereomers were separated by SFC (Column: Welk-01(R,R)(250×4.6) mm, 5 μm; %CO2: 45%; % co-solvent: 5 mM ammonium acetate / methanol and acetonitrile (1:1); Flow rate: 4 g / min; Temperature: 30° C.; UV: 237 nm) and the first isomeric fraction eluting with a retention time of 2.28 min was concentrated to dryness and lyophilized to give 2-amino-6-((3S)-2-(2,6-dioxopiperidin-3-yl)-3-methyl-1-oxoisoindolin-5-yl)-4-methylnicotinonitrile (20 mg, 12% yield). LCMS (Method D): Retention time 1.64 min. [M+H] + 390.0; 1H NMR (400 MHz, DMSO-d6) δ 11.01-10.72 (s, 1H), 8.28 (s, 1H), 8.19 (dd, J=8.0, 1.3 Hz, 1H), 7.76 (d, J=8.0 Hz, 1H), 7.35 (s, 1H), 6.92 (s, 2H), 4.94-4.68 (m, 2H), 2.65-2.58 (m, 2H), 2.44 (s, 3H), 2.14-1.91 (m, 2H), 1.51 (d, J=6.6 Hz, 3H); and the second isomeric fraction eluting at a retention time of 3.25 min were concentrated to dryness and lyophilized to give 2-amino-6-((3R)-2-(2,6-dioxopiperidin-3-yl)-3-methyl-1-oxoisoindolin-5-yl)-4-methylnicotinonitrile (20 mg, 12% yield). LCMS (Method D): Retention time 1.66 min, [M+H] + 390.0; 1 H NMR (400 MHz, DMSO-d6) δ 11.01-10.72 (s, 1H), 8.27 (s, 1H), 8.20 (d, J=8.0 Hz, 1H), 7.78 (d, J=8.0 Hz, 1H), 7.34 (s, 1H), 6.90 (s, 2H), 4.84-4.70 (m, 2H), 2.84-2.67 (m, 2H), 2.44 (s, 3H), 2.14-1.91 (m, 2H), 1.49 (br d, J=6.6 Hz, 3H).

[0265] Examples 12 and 13 3-((S)-5-(4,5-dimethyl-6-(methylamino)pyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione and 3-((R)-5-(4,5-dimethyl-6-(methylamino)pyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione [ka] Intermediates 12A and 13A: To a solution of intermediate 8D (350 mg, 0.764 mmol) in dioxane (8 mL) was added 6-chloro-N,3,4-trimethylpyridin-2-amine (130 mg, 0.764 mmol), followed by 3 M aqueous potassium phosphate (0.764 mL, 2.291 mmol). The reaction mixture was purged with nitrogen at room temperature for 15 minutes. PdCl2(dppf)-CH2Cl2 adduct (62.4 mg, 0.076 mmol) was added under nitrogen and the reaction mixture was heated at 100 °C for 12 hours. The reaction mixture was cooled to room temperature, filtered through a pad of Celite, and concentrated under reduced pressure. The residue was purified by flash chromatography (SiO2, 24 g column, 0-10% MeOHDCM) to give tert-butyl (4S)-5-amino-4-(5-(4,5-dimethyl-6-(methylamino)pyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)-5-oxopentanoate (230 mg, 0.493 mmol, 64.6% yield) as an off-white solid. LCMS (Method A): Retention time 1.81 min, [M+H] + 467.4.

[0266] Examples 12 and 13: To a stirred solution of tert-butyl 5-amino-4-(5-(4,5-dimethyl-6-(methylamino)pyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)-5-oxopentanoate (200 mg, 0.429 mmol) in acetic acid (1 mL) was added benzenesulfonic acid (67.8 mg, 0.429 mmol). The reaction mixture was heated to 120° C. under microwave irradiation for 30 min. The volatiles were removed under reduced pressure and the resulting crude product was purified by preparative HPLC (Column: X Select CSH C18 (250×20 mm) 5 μm Mobile phase: A: 10 mM ammonium acetate B: CAN T / B: 0 / 20, 18 / 85, 20 / 100, 21 / 20, 23 / 20; Flow rate: 20 mL / min). The desired fractions were lyophilized to give 3-(5-(4,5-dimethyl-6-(methylamino)pyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione as a diastereomeric mixture. LCMS (Method A): Retention time: 1.47 min, [M+H]+ 393.4. The diastereomers were separated by SFC (column: Chiralpak IC (250 × 4.5 μm; % CO2: 45%; % co-solvent: 5 mM ammonium acetate / methanol and acetonitrile (1:1); flow rate: 4 g / min; temperature: 30 °C; UV: 237 nm) and the fraction of the first eluting isomer (eluting at retention time: 8.65 min) was concentrated to dryness and lyophilized to give 3-((S)-5-(4,5-dimethyl-6-(methylamino)pyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (3 mg, 2% yield): LCMS (Method D): retention time 1.743 min, [M+H] + 393.4; 1H NMR (400 MHz, DMSO-d6) δ 11.07-10.81 (m, 1H), 8.35-8.12 (m, 2H), 7.71 (d, J=8.0 Hz, 1H), 7.14 (s, 1H), 6.03 (br d, J=4.0 Hz, 1H), 4.88-4.69 (m, 2H), 2.96 (d, J=4.5 Hz, 3H), 2.78-2.63 (m, 3H), 2.26 (s, 3H), 2.03 (s, 4H), 1.49 (d, J=6.5 Hz, 3H), the fractions of the second peak eluting at retention time 11.18 min were concentrated and lyophilized to give 3-((R)-5-(4,5-dimethyl-6-(methylamino)pyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (3 mg, 2% yield). LCMS (Method D): Retention time 1.669 min, [M+H] + 393.4; 1 H NMR (400 MHz, DMSO-d6) δ 11.07-10.81 (m, 1H), 8.35-8.12 (m, 2H), 7.71 (d, J=8.0 Hz, 1H), 7.14 (s, 1H), 6.03 (br d, J=4.0 Hz, 1H), 4.88-4.69 (m, 2H), 2.96 (d, J=4.5 Hz, 3H), 2.78-2.63 (m, 3H), 2.26 (s, 3H), 2.03 (s, 4H), 1.49 (d, J=6.5 Hz, 3H).

[0267] Examples 14 and 15 6-((3S)-2-(2,6-dioxopiperidin-3-yl)-3-methyl-1-oxoisoindolin-5-yl)-4-methyl-2-(methylamino)nicotinonitrile and 6-((3R)-2-(2,6-dioxopiperidin-3-yl)-3-methyl-1-oxoisoindolin-5-yl)-4-methyl-2-(methylamino)nicotinonitrile [ka]

[0268] Intermediates 14A and 15A: To a solution of intermediate 8D (200 mg, 0.436 mmol) in dioxane (8 mL) was added 6-chloro-4-methyl-2-(methylamino)nicotinonitrile (79 mg, 0.436 mmol) followed by K3PO4 (0.291 mL, 0.873 mmol 3 M aqueous solution). The reaction mixture was purged with nitrogen at room temperature for 15 minutes. Xphos Pd G4 (37.6 mg, 0.044 mmol) was added under nitrogen atmosphere and the reaction mixture was heated at 80° C. for 12 hours. The reaction mixture was cooled to room temperature, filtered through a celite pad and concentrated under reduced pressure. The residue was purified by flash chromatography (SiO2, 24 g column, 0-10% MeOHDCM) to give tert-butyl (4S)-5-amino-4-(5-(cyano-4-methyl-6-(methylamino)pyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)-5-oxopentanoate (75 mg, 0.157 mmol, 36.0% yield) as an off-white solid. LCMS (Method A): Retention time 1.59 min, [M+H] + 478.2.

[0269] Examples 14 and 15: To a solution of tert-butyl (4S)-5-amino-4-(5-(5-cyano-4-methyl-6-(methylamino)pyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)-5-oxopentanoate (60 mg, 0.126 mmol) in acetonitrile (3 mL) at room temperature was added benzenesulfonic acid (19.87 mg, 0.126 mmol). The reaction mixture was heated to 120° C. in a microwave reactor for 45 minutes. The reaction mixture was cooled to room temperature and concentrated under reduced pressure to give the crude product. The crude product was purified by flash chromatography (SiO2, 24 g column, 0-10% MeOH / DCM) to give a diastereomeric mixture of 6-(2-(2,6-dioxopiperidin-3-yl)-3-methyl-1-oxoisoindolin-5-yl)-4-methyl-2-(methylamino)nicotinonitrile as an off-white solid. LCMS (Method A): Retention time 1.22 min, [M+H]+ 404.1. The diastereomers were separated by SFC (Column Welk-01(R,R)(250*4.6)mm, 5 μm; % CO2: 45%; % co-solvent: 5 mM ammonium acetate / methanol and acetonitrile (1:1); flow rate: 4 g / min; temperature 30°C; UV: 237 nm) and the first isomeric fraction eluting with a retention time of 2.91 min was concentrated to dryness and lyophilized to give 6-((3S)-2-(2,6-dioxopiperidin-3-yl)-3-methyl-1-oxoisoindolin-5-yl)-4-methyl-2-(methylamino)nicotinonitrile (6 mg, 12% yield): LCMS (Method D): retention time 2.071 min, [M+H] + 404.2; 1H NMR(400 MHz, DMSO-d6) δ 11.27-10.74 (m, 1H), 8.35 (s, 1H), 8.28 (d, J=8.0 Hz, 1H), 7.77 (d, J=8.5 Hz, 1H), 7.37 (s, 1H), 7.12 (br d, J=4.5 Hz, 1H), 4.87 (q, J=6.2 Hz, 1H), 4.81-4.71 (m, 1H), 3.00 (d, J=4.5 Hz, 3H), 2.68 (s, 1H), 2.44 (s, 3H), 1.72 (s, 3H), 1.52 (d, J=7.0 Hz, 3H), retention time 4.74 The fractions of the second peak eluting at 2.0 min were concentrated and lyophilized to give 6-((3R)-2-(2,6-dioxopiperidin-3-yl)-3-methyl-1-oxoisoindolin-5-yl)-4-methyl-2-(methylamino)nicotinonitrile (8 mg, 16% yield). LCMS (Method D): Retention time 2.063 min, [M+H] + 404.2;1H NMR (400 MHz, DMSO-d6) δ 11.08-10.87 (m, 1H), 8.34 (s, 1H), 8.28 (d, J=8.3 Hz, 1H), 7.77 (d, J=8.0 Hz, 1H), 7.36 (s, 1H), 7.19-7.05 (m, 1H), 4.91-4.71 (m, 2H), 2.99 (d, J=4.5 Hz, 3H), 2.76-2.57 (m, 3H), 2.44 (s, 3H), 1.71 (s, 1H), 1.49 (d, J=6.8 Hz, 3H).

[0270] Examples 16 and 17 3-((S)-5-(6-amino-4,5-dimethylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione and 3-((R)-5-(6-amino-4,5-dimethylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione [ka]

[0271] Intermediate 16A: 4-Bromo-2-ethyl-3-fluorobenzoic acid A solution of 4-bromo-2,3-difluorobenzoic acid (2.0 g, 8.44 mmol) in tetrahydrofuran (40 mL) was cooled to -78 °C and a 1 M solution of ethylmagnesium bromide (8.44 mL, 25.3 mmol) in THF was added dropwise. The reaction mixture was allowed to warm to room temperature and stirred under nitrogen for 12 h. MeOH (15 mL) was added dropwise at 0 °C to quench the reaction. The volatiles were removed under reduced pressure and the residue was partitioned between EtOAc and 2 M aqueous hydrochloric acid. The layers were separated and extracted with EtOAc. The combined organic layers were washed with brine, dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give the crude product. The crude product was purified by flash chromatography (SiO2, 40 g column, 0-80% EtOAc / petroleum ether) to give 4-bromo-2-ethyl-3-fluorobenzoic acid (1 g, 48% yield). LCMS (Method A): Retention time 0.69 min; [MH] + 245.1, 247.1; 1 H NMR (300 MHz, DMSO-d6) δ 13.36 (s, 1H), 7.68-7.53 (m, 2H), 2.95 (qd, J = 7.4, 2.6 Hz, 2H), 1.24-1.06 (m, 3H).

[0272] Intermediate 16B: Methyl 4-bromo-2-ethyl-3-fluorobenzoate To a stirred mixture of 4-bromo-2-ethyl-3-fluorobenzoic acid (0.7 g, 2.83 mmol) and K2CO3 (0.783 g, 5.67 mmol) in acetone (15 mL) was added dimethyl sulfate (0.541 mL, 5.67 mmol) dropwise at room temperature. The reaction mixture was stirred at 50 °C for 14 h and filtered through a pad of Celite. The filtrate was concentrated in vacuo and purified by flash chromatography (SiO2, 24 g column, 0-50% EtOAc / petroleum ether) to give methyl 4-bromo-2-ethyl-3-fluorobenzoate (0.51 g, 69% yield) as a colorless oil. 1 H NMR (300 MHz, CHLOROFORM-d) δ 7.58-7.51 (m, 1H), 7.49-7.37 (m, 1H), 3.90 (s, 3H), 3.01 (qd, J = 7.4, 2.6 Hz, 2H), 1.23 (t, J = 7.4 Hz, 3H).

[0273] Intermediate 16C: Methyl 4-bromo-2-(1-bromoethyl)-3-fluorobenzoate To a stirred solution of methyl 4-bromo-2-ethyl-3-fluorobenzoate (0.515 g, 1.972 mmol) in DCE (10 mL) was added NBS (0.386 g, 2.170 mmol) followed by AIBN (0.065 g, 0.394 mmol). The reaction mixture was heated at 85° C. for 15 h. The reaction mixture was diluted with ethyl acetate and washed with saturated 10% sodium thiosulfate solution and brine solution. The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The resulting residue was purified by flash chromatography (SiO2, 24 g column, 0-30% EtOAc / petroleum ether) to give methyl 4-bromo-2-(1-bromoethyl)-3-fluorobenzoate (0.6 g, 89% yield) as a white solid. 1H NMR (300 MHz, CHLOROFORM-d) δ 7.70-7.53 (m, 1H), 7.52-7.44 (m, 1H), 6.16-5.87 (m, 1H), 3.93 (s, 3H), 1.95 (dd, J = 7.0, 1.3 Hz, 3H).

[0274] Intermediate 16D: tert-Butyl (4S)-5-amino-4-(5-bromo-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)-5-oxopentanoate To a stirred solution of methyl 4-bromo-2-(1-bromoethyl)-3-fluorobenzoate (0.86 g, 2.53 mmol) and H-Glu(OtBu)-NH2HCl (0.845 g, 3.54 mmol) in acetonitrile (15 mL) was added DIPEA (1.325 mL, 7.59 mmol). The reaction mixture was heated at 85 °C for 15 h. The volatiles were removed under reduced pressure and purified by flash chromatography (SiO2, 40 g column, 0-10% MeOH / DCM) to give tert-butyl (4S)-5-amino-4-(5-bromo-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)-5-oxopentanoate (0.23 g, 21% yield) as a pale yellow solid. LCMS (Method A): Retention time 1.19 min, [M+23H] + 451.3, 453.4.

[0275] Intermediate 16E: tert-butyl (4S)-5-amino-4-(5-(6-amino-4,5-dimethylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)-5-oxopentanoate To a stirred solution of tert-butyl (4S)-5-amino-4-(5-bromo-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)-5-oxopentanoate (350 mg, 0.815 mmol) in 1,4-dioxane (20 mL) was added 3,4-dimethyl-6-(trimethylstannyl)pyridin-2-amine (232 mg, 0.815 mmol). The mixture was purged with argon for 5 minutes and bis(triphenylphosphine)palladium(II) chloride (57.2 mg, 0.082 mmol) was added. The reaction mixture was heated at 100° C. for 16 hours, cooled to room temperature and diluted with ethyl acetate. The mixture was washed with brine solution and the organic layer was dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give the crude compound. The crude compound was purified by flash chromatography (SiO2, 24 g column, 0-5% MeOH / DCM) and the combined product fractions were concentrated in vacuo to give tert-butyl (4S)-5-amino-4-(5-(6-amino-4,5-dimethylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)-5-oxopentanoate (270 mg, 70.4% yield) as a light brown solid. LCMS (Method A): Retention time 1.64 min; [M+H] + 471.2 1 H NMR (400 MHz, DMSO-d6) δ 7.99 (t, J=6.9 Hz, 1H), 7.59 (s, 1H), 7.47 (br s, 1H), 7.30-7.13 (m, 2H), 6.90 (s, 1H), 5.74 (br d, J=5.3 Hz, 2H), 4.89 (q, J=6.1 Hz, 1H), 4.50-4.37 (m, 1H), 2.31-2.19 (m, 6H), 2.04 (s, 3H), 1.55-1.45 (m, 3H), 1.37 (d, J=2.9 Hz, 9H).

[0276] Examples 16 and 17: To a stirred solution of tert-butyl (4S)-5-amino-4-(5-(6-amino-4,5-dimethylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)-5-oxopentanoate (270 mg, 0.574 mmol) in acetonitrile (10 mL) was added PTSOH (218 mg, 1.148 mmol). The reaction mixture was heated under microwave irradiation at 120° C. for 30 min. Volatiles were removed under reduced pressure and the resulting crude product was purified by prep-HPLC using a polar organic solvent method (cellulose-2 [250 x 4.6 mm], 10 mM ammonium acetate / MeOH, flow rate: 23 mL / min (isocratic gradient)), the first isomeric fraction with retention time 9.73 min was concentrated to dryness and lyophilized to give 3-((S)-5-(6-amino-4,5-dimethylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (60 mg, 27% yield). LCMS (Method G): retention time 1.83 min; [M+H] + 397.1; 1 H NMR (400 MHz, DMSO-d6) δ 11.10-10.87 (m, 1H), 8.01 (t, J=7.3 Hz, 1H), 7.57 (d, J=8.0 Hz, 1H), 6.92 (d, J=2.0 Hz, 1H), 5.78 (s, 2H), 5.01 (q, J=6.5 Hz, 1H), 4.73 (br dd, J=12.3, 4.8 Hz, 1H), 2.82-2.67 (m, 1H), 2.64-2.55 (m, 2H), 2.23 (s, 3H), 2.04 (s, 3H), 1.54 (d, J=7.0 Hz, 3H). The fractions of the second peak eluting at a retention time of 13.96 min were concentrated to dryness and lyophilized to give 3-((R)-5-(6-amino-4,5-dimethylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (70 mg, 31% yield). LCMS (Method G): Retention time 1.84 min; [M+H] + 397.1; 1H NMR (400 MHz, DMSO-d6) δ 11.07-10.89 (m, 1H), 8.01 (t, J=7.1 Hz, 1H), 7.58 (d, J=8.0 Hz, 1H), 6.91 (s, 1H), 5.79 (s, 2H), 4.89 (q, J=6.5 Hz, 1H), 4.80 (dd, J=12.6, 5.1 Hz, 1H), 2.94-2.78 (m, 1H), 2.73-2.59 (m, 2H), 2.22 (s, 3H), 2.03 (s, 3H), 1.51 (d, J=6.8 Hz, 3H).

[0277] Method A: ACQUITY UPLC® BEH C18 (3.0 x 50 mm), 1.7 μm; Mobile phase A: 95:5 water:acetonitrile (containing 2.5 mM NH4OAc); Mobile phase B: 5:95 water:acetonitrile (containing 2.5 mM NH4OAc); Temperature: 40 °C; Gradient: 20% B to 100% B over 2 min; Flow rate: 0.7 mL / min; Detection: MS and UV (220 nm). Method B: Column: XBridge BEH XP C18 (50 x 2.1) mm, 2.5 μm; Mobile phase A: 95:5 water:acetonitrile (containing 10 mM NH4OAc); Mobile phase B: 5:95 water:acetonitrile (containing 10 mM NH4OAc); Temperature: 50 °C; Gradient: 0% B to 100% B over 3 min; Flow rate: 1.1 mL / min; Detection: MS and UV (220 nm). Method D: Column- Kinetex XB-C18 (75 x 3 mm-2.6 μm); Mobile phase A: 5 mM NH4COOH / water; Mobile phase B: Acetonitrile; Gradient: 10% B to 50% B over 3 min; Flow rate: 1.0 mL / min; 50% B to 100% B until 4.1 min; Flow rate: 1.0 mL / min; Hold until 4.5 min; 90% B from 4.5 min to 5.0 min; Flow rate: 1.5 mL / min; Detection: MS and UV (220 nm). Method G: Column- Kinetex XB-C18 (75 x 3 mm-2.6 μm); Mobile phase A: 5 mM NH4CO2H / water; Mobile phase B: Acetonitrile; Gradient: 20% B to 100% B over 4 min; Flow rate: 1.0 mL / min; Hold until 4.6 min; Flow rate: 1.5 mL / min; Hold until 4.7 min; 20% B from 4.7 min to 5.0 min; Flow rate: 1.0 mL / min; Detection: MS and UV (220 nm).

[0278] Example 18 3-(5-(6-amino-3-methylpyridin-2-yl)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione [ka] Example 18 was synthesized from 6-chloropyridin-2-amine and intermediate 3B using general method 2. The crude product was purified by preparative LCMS (Column: Waters XBridge C18, 150 mm x 19 mm, 5 μm particle size; Mobile phase A: 5:95 acetonitrile:water (with 0.1% trifluoroacetic acid); Mobile phase B: 95:5 acetonitrile:water (with 0.1% trifluoroacetic acid); Gradient: 10% B hold for 0 min, 10-30% B over 20 min, then 100% B hold for 5 min; Flow rate: 20 mL / min. LCMS (Method B): Retention time 0.98 min, [M+H] + 369.1; 1 H NMR (400 MHz, DMSO-d6) δ 11.04 (s, 1H), 7.77 (br d, J = 7.6 Hz, 2H), 7.74-7.68 (m, 1H), 6.93-6.87 (m, 1H), 5.17 (dd, J = 13.3, 5.1 Hz, 1H), 4.67 (d, J = 17.6 Hz, 1H), 4.55-4.47 (m, 1H), 3.01-2.89 (m, 2H), 2.67-2.61 (m, 1H), 2.47-2.39 (m, 1H), 2.04 (s, 3H).

[0279] General method 4: A mixture of aryl halide (1 eq.), arylboronic acid pinacol ester (1.0 eq.), potassium carbonate (1.5 eq.), dioxane (4 mL / mmol) and water (0.4 mL / mmol) was purged with argon at room temperature for 5 min. [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride dichloromethane complex (0.05 eq.) was added and the reaction mixture was heated at 110 °C for 2 h. The reaction mixture was cooled at room temperature, diluted with EtOAc and filtered through a Celite pad. The filtrate was washed with brine solution, dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by flash chromatography. The isolated product was dissolved in acetonitrile, pTSA·H2O (2 eq.) was added and the mixture was heated at 120 °C for 1.5 h in a microwave reactor. The reaction mixture was cooled to room temperature, concentrated in vacuo and the crude product was purified by prep-HPLC to give the desired product.

[0280] Example 19 3-(5-(6-amino-5-fluoro-4-methylpyridin-2-yl)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione [ka] Intermediates 19A and 19B: 6-bromo-3-fluoro-4-methylpyridin-2-amine and 6-bromo-5-fluoro-4-methylpyridin-2-amine To a solution of 6-bromo-4-methylpyridin-2-amine (2.6 g, 13.90 mmol) in chloroform (100 mL) and water (100 mL) was added Selectfluor (2.462 g, 6.95 mmol). The reaction mixture was stirred at room temperature for 48 h. The reaction mixture was diluted with DCM (200 mL), washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The resulting residue was purified by flash chromatography (SiO2, 80 g column, 0~30% EtOAc / petroleum ether) to give Intermediate 19A: 6-bromo-3-fluoro-4-methylpyridin-2-amine (600 mg, 19.6% yield); LCMS (Method A): Retention time 1.21 min, [M+H] + 207.1. 1 H NMR (300 MHz, DMSO-d6) δ 6.60 (d, J = 4.2 Hz, 1H), 6.55 (s, 2H), 2.13 (d, J = 1.9 Hz, 3H); and Intermediate 19B: 6-Bromo-5-fluoro-4-methylpyridin-2-amine (500 mg, 4.4% yield); LCMS (Method A): retention time 1.14, [M+H] + 207.1; 1 H NMR (300 MHz, chloroform-d) δ 6.25 (d, J = 4.2 Hz, 1H), 4.52-4.22 (m, 2H), 2.23 (d, J = 1.1 Hz, 3H).

[0281] Example 19: Example 19 was synthesized by using general method 4 using 6-bromo-3-fluoro-4-methylpyridin-2-amine and tert-butyl (S)-5-amino-4-(4-fluoro-1-oxo-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoindolin-2-yl)-5-oxopentanoate (synthesized by the method shown in Example 2 starting from 4-bromo-3-fluoro-2-methylbenzoic acid). The crude product was purified by preparative LCMS (Column: YMC EXRS 250 mm x 21 mm, Mobile phase A: 5:95 acetonitrile:water (with 0.1% trifluoroacetic acid); Mobile phase B: 95:5 acetonitrile:water (with 0.1% trifluoroacetic acid); Gradient: 10% B hold for 0 min, 10-30% B over 20 min, then 100% B hold for 5 min; Flow rate: 20 mL / min. LCMS (Method D): Retention time 1.56 min; [M+H] + 387.15; 1 H NMR (400 MHz, DMSO-d6) δ 11.02 (s, 1H), 8.09-7.92 (m, 1H), 7.66 (d, J = 8.0 Hz, 1H), 7.32-6.78 (m, 1H), 5.14(dd, J = 13.8, 5.8 Hz,1H), 4.74-4.33 (m, 2H), 2.98-2.86 (m, 1H), 2.68-2.56 (m, 2H), 2.24 (s, 3H), 22.09-2.05(m, 1H).

[0282] Example 20 3-(5-(6-amino-3-fluoro-4-methylpyridin-2-yl)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione [ka] Example 20 was synthesized using general method 4 using 6-bromo-5-fluoro-4-methylpyridin-2-amine and tert-butyl (S)-5-amino-4-(4-fluoro-1-oxo-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoindolin-2-yl)-5-oxopentanoate (synthesized by the method set forth in Example 2 starting with 4-bromo-3-fluoro-2-methylbenzoic acid). The crude product was purified by preparative LCMS (Column: Waters XBridge C18, 150 mm x 19 mm, 5 μm particle size; Mobile phase A: 5:95 acetonitrile:water (with 0.1% trifluoroacetic acid); Mobile phase B: 95:5 acetonitrile:water (with 0.1% trifluoroacetic acid); Gradient: 10% B hold for 0 min, 10-30% B over 20 min, then 100% B hold for 5 min; Flow rate: 20 mL / min. LCMS (Method B): Retention time 1.03 min, [M+H] + 387.1; 1 H NMR (400 MHz, DMSO-d6) δ 11.04 (s, 1H), 7.74-7.57 (m, 2H), 6.46 (d, J = 4.5 Hz, 1H), 5.97 (s, 2H), 5.16 (dd, J = 5.0, 13.3 Hz, 1H), 4.65 (d, J = 17.5Hz, 1H), 4.52-4.42 (m, 1H), 3.03-2.86 (m, 1H), 2.68-2.59 (m, 1H), 2.49-2.41 (m, 1H), 2.21 (s, 3H), 2.12-1.99 (m, 1H).

[0283] Examples 21 and 22 3-(5-(6-amino-5-fluoro-4-methylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione [ka] Intermediate 21A: tert-butyl (4S)-5-amino-4-(3-methyl-1-oxo-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoindolin-2-yl)-5-oxopentanoate A mixture of intermediate 8D (1.0 g, 2.43 mmol), potassium acetate (0.239 g, 2.43 mmol) and Bispin (0.617 g, 2.43 mmol) in anhydrous DME (15 mL) was purged with argon gas for 10 min at room temperature. PdCl2(dppf)-CH2Cl2 adduct (0.159 g, 0.195 mmol) was added under an argon atmosphere. The vial was sealed and the mixture was heated at 90 °C for 12 h. The reaction mixture was cooled at room temperature, filtered through a celite pad and the filtrate was concentrated under reduced pressure. The residue was dissolved in diethyl ether and filtered through a celite pad. The filtrate was concentrated under reduced pressure to give tert-butyl (4S)-5-amino-4-(3-methyl-1-oxo-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoindolin-2-yl)-5-oxopentanoate (1.0 g, 90% yield). LCMS (Method A): Retention time 1.70 min, [M+H] + 459.1.

[0284] Preparation of 21B and 22B: tert-butyl (4S)-5-amino-4-(5-(6-amino-5-fluoro-4-methylpyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)-5-oxopentanoate To a solution of tert-butyl (4S)-5-amino-4-(3-methyl-1-oxo-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoindolin-2-yl)-5-oxopentanoate (0.241 g, 0.527 mmol) in dioxane (10 mL) was added 6-bromo-3-fluoro-4-methylpyridin-2-amine (0.09 g, 0.44 mmol) followed by sodium bicarbonate (0.5 M solution, 2.195 mL, 1.097 mmol). The reaction mixture was purged with nitrogen for 15 minutes at room temperature and bis(triphenylphosphine)palladium(II) chloride (0.031 g, 0.044 mmol) was added under nitrogen and heated at 100° C. for 12 hours. The reaction mixture was cooled to room temperature, filtered through a pad of Celite, and concentrated under reduced pressure. The residue was purified by flash chromatography (SiO2, 24 g column, 50-100% EtOAc / DCM) to give tert-butyl (4S)-5-amino-4-(5-(6-amino-5-fluoro-4-methylpyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)-5-oxopentanoate (150 mg) as a diastereomeric mixture. The diastereomers were separated by SFC (column Chiral Pak IG (250×4.6) mm, 5 μm; %CO2: 45%; %cosolvent: 5 mM ammonium acetate / methanol and acetonitrile (1:1); flow rate: 4 g / min; temperature: 30° C.; UV: 237 nm) and the first isomeric fraction eluting with a retention time of 3.4 min was concentrated to dryness to give tert-butyl (4S)-5-amino-4-(5-(6-amino-5-fluoro-4-methylpyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)-5-oxopentanoate (40 mg, 20% yield). LCMS (Method A): Retention time 1.41, [M+H] +457.1. Fractions of the second peak eluting at a retention time of 4.6 minutes were concentrated to give tert-butyl (4S)-5-amino-4-(5-(6-amino-5-fluoro-4-methylpyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)-5-oxopentanoate (50 mg, 25% yield). LCMS (Method A): Retention time 1.40, [M+H] + 457.4.

[0285] Example 21: To a stirred solution of (4S)-5-amino-4-(5-(6-amino-5-fluoro-4-methylpyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)-5-oxopentanoate (0.04 g, 0.088 mmol) in acetonitrile (10 mL) was added benzenesulfonic acid (0.028 g, 0.175 mmol). The reaction mixture was heated at 120° C. for 30 min under microwave irradiation. The reaction mixture was cooled to room temperature, concentrated under reduced pressure and purified by preparative LCMS (Column: Waters XBridge C18, 150 mm x 19 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile:water (with 0.1% trifluoroacetic acid); Mobile phase B: 95:5 acetonitrile:water (with 0.1% trifluoroacetic acid); Gradient: 10% B hold for 0 min, 10-30% B over 20 min, then 100% B hold for 5 min; Flow rate: 20 mL / min. LCMS (Method B): Retention time 1.19 min, [M+H] + 383.1; 1 H NMR (400 MHz, DMSO-d6) δ 10.95 (s, 1H), 8.14 (s, 1H), 8.07 (dd, J = 1.1, 8.1 Hz, 1H), 7.72 (d, J = 8.0 Hz, 1H), 7.17 (d, J = 4.4 Hz, 1H),4.82-4.67 (m, 2H), 2.93-2.79 (m, 1H), 2.71-2.59 (m, 2H), 2.26 (s, 3H), 2.05-1.96 (m, 1H), 1.48 (d, J = 6.6 Hz, 3H).

[0286] Example 22: To a stirred solution of (4S)-5-amino-4-(5-(6-amino-5-fluoro-4-methylpyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)-5-oxopentanoate (0.04 g, 0.088 mmol) in acetonitrile (10 mL) was added benzenesulfonic acid (0.028 g, 0.175 mmol). The reaction mixture was heated at 120° C. for 30 min under microwave irradiation. The reaction mixture was cooled at room temperature, concentrated under reduced pressure and the residue was purified by preparative LCMS (Column: Waters XBridge C18, 150 mm x 19 mm, 5 μm particle size; Mobile phase A: 5:95 acetonitrile:water (with 0.1% trifluoroacetic acid); Mobile phase B: 95:5 acetonitrile:water (with 0.1% trifluoroacetic acid); Gradient: 10% B hold for 0 min, 10-30% B over 20 min, then 100% B hold for 5 min; Flow rate: 20 mL / min. LCMS (Method B): Retention time 1.19 min, [M+H] + 383.1; 1 H NMR (400 MHz, DMSO-d6) δ 10.92 (s, 1H), 8.16 (s, 1H), 8.07 (dd, J = 1.1, 8.0 Hz, 1H), 7.70 (d, J = 8.0 Hz, 1H), 7.17 (d, J = 4.3 Hz, 1H),6.26 (br s, 2H), 4.83 (q, J = 6.6 Hz, 1H), 4.74 (br dd, J = 4.3, 11.3 Hz, 1H), 2.83-2.71 (m, 1H), 2.64-2.55 (m, 2H), 2.25 (d, J = 1.5 Hz, 3H), 2.09 -1.96 (m, 1H), 1.50 (d, J = 6.6 Hz, 3H).

[0287] Example 23 2-Amino-6-((3R)-2-(2,6-dioxopiperidin-3-yl)-4-fluoro-3-methyl-1-oxoisoindolin-5-yl)-4-methylnicotinonitrile [ka]

[0288] Example 23 was synthesized using general method 4 from 2-amino-6-chloro-4-methylnicotinonitrile and tert-butyl (S)-5-amino-4-((R)-4-fluoro-3-methyl-1-oxo-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoindolin-2-yl)-5-oxopentanoate (synthesized by the method set forth in Example 2 starting with 4-bromo-3-fluoro-2-methylbenzoic acid). The crude product was purified by preparative LCMS (Column: Waters XBridge C18, 150 mm x 19 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile:water (with 0.1% trifluoroacetic acid); Mobile phase B: 95:5 acetonitrile:water (with 0.1% trifluoroacetic acid); Gradient: 10% B hold for 0 min, 10-30% B over 20 min, then 100% B hold for 5 min; Flow rate: 20 mL / min. LCMS (Method B): Retention time 1.19 min, [M+H] + 408.0; 1 H NMR (400 MHz, DMSO-d6) δ 11.00 (s, 1H), 8.17-7.85 (m, 1H), 7.73-7.52 (m, 1H), 7.05 (s, 1H), 6.99 (s, 2H), 5.08-4.89 (m, 1H), 4.86-4.72 (m, 1H), 2.90-2.75 (m, 1H), 2.74-2.58 (m, 2H), 2.42 (s, 3H), 2.12-2.00 (m, 1H), 1.57-1.47 (m, 3H).

[0289] Example 24 3-((R)-5-(6-amino-4-methylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione [ka] Example 24 was synthesized using general method 4 from 6-chloro-4-methylpyridin-2-amine and tert-butyl (S)-5-amino-4-((R)-4-fluoro-3-methyl-1-oxo-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoindolin-2-yl)-5-oxopentanoate (synthesized by the method set forth in Example 2 starting with 4-bromo-3-fluoro-2-methylbenzoic acid). The crude product was purified by preparative LCMS (Column: Waters XBridge C18, 150 mm x 19 mm, particle size 5 μm; Mobile phase A: 5:95 acetonitrile:water (with 0.1% trifluoroacetic acid); Mobile phase B: 95:5 acetonitrile:water (with 0.1% trifluoroacetic acid); Gradient: 10% B hold for 0 min, 10-30% B over 20 min, then 100% B hold for 5 min; Flow rate: 20 mL / min. LCMS (Method B): Retention time 1.20 min, [M+H] + 383.2; 1 H NMR (400 MHz, DMSO-d6) δ 11.00 (d, J = 0.8 Hz, 1H), 8.01-7.84 (m, 1H), 7.71 (d, J = 7.8 Hz, 1H), 6.96 (br d, J = 1.0 Hz, 1H), 6.71-6.54 (m, 1H), 4.99-4.91 (m, 1H), 4.87-4.78 (m, 1H), 2.86 (br dd, J = 4.4, 3.4 Hz, 1H), 2.75-2.62 (m, 2H), 2.47-2.44 (m, 1H), 2.34 (s, 4H), 1.94 (d, J = 16.0 Hz, 1H), 1.53 (d, J = 6.8 Hz, 3H).

[0290] Example 25 3-((R)-5-(6-amino-4-(trifluoromethyl)pyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione [ka] Example 25 was synthesized using general method 4 from 6-chloro-4-(trifluoromethyl)pyridin-2-amine and tert-butyl (S)-5-amino-4-((R)-4-fluoro-3-methyl-1-oxo-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoindolin-2-yl)-5-oxopentanoate (synthesized by the method set forth in Example 2 starting with 4-bromo-3-fluoro-2-methylbenzoic acid). The crude product was purified by preparative LCMS (Column: Waters XBridge C18, 150 mm x 19 mm, 5 μm particle size; Mobile phase A: 5:95 acetonitrile:water (with 0.1% trifluoroacetic acid); Mobile phase B: 95:5 acetonitrile:water (with 0.1% trifluoroacetic acid); Gradient: 10% B hold for 0 min, 10-30% B over 20 min, then 100% B hold for 5 min; Flow rate: 20 mL / min. LCMS (Method B): Retention time 1.39 min, [M+H] + 438.1; 1 H NMR (400 MHz, DMSO-d6) δ 10.99 (s, 1H), 8.13-7.95 (m, 1H), 7.65 (d, J = 7.8 Hz, 1H), 7.15 (s, 1H), 6.79 (s, 2H), 4.98-4.88 (m, 1H), 4.86-4.77 (m, 1H), 2.85 (br s, 1H), 2.93-2.78 (m, 1H), 2.72-2.59 (m, 2H), 2.08-2.00 (m, 1H), 1.52 (d, J = 6.8 Hz, 3H).

[0291] Example 26 3-((R)-5-(6-amino-3-methylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione [ka] Example 26 was synthesized using general method 4 from 6-chloro-5-methylpyridin-2-amine and tert-butyl (S)-5-amino-4-((R)-4-fluoro-3-methyl-1-oxo-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoindolin-2-yl)-5-oxopentanoate (synthesized by the method set forth in Example 2 starting with 4-bromo-3-fluoro-2-methylbenzoic acid). The crude product was purified by preparative LCMS (Column: Waters XBridge C18, 150 mm x 19 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile:water (with 0.1% trifluoroacetic acid); Mobile phase B: 95:5 acetonitrile:water (with 0.1% trifluoroacetic acid); Gradient: 10% B hold for 0 min, 10-30% B over 20 min, then 100% B hold for 5 min; Flow rate: 20 mL / min. LCMS (Method B): Retention time 1.07 min, [M+H] + 383.1; 1 H NMR (400 MHz, DMSO-d6) δ 11.01 (s, 1H), 7.90-7.77 (m, 1H), 7.74-7.61 (m, 2H), 6.86 (br dd, J = 5.5, 3.9 Hz, 1H), 4.96 (q, J = 6.5 Hz, 1H), 4.84 (dd, J = 12.6, 5.1 Hz, 1H), 3.01-2.82 (m, 2H), 2.71-2.60 (m, 2H), 2.12-2.01 (m, 5H), 1.51 (d, J = 6.6 Hz, 3H).

[0292] Example 27 3-((R)-5-(6-aminopyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione [ka] Example 27 was synthesized using general method 4 from 6-chloropyridin-2-amine and tert-butyl (S)-5-amino-4-((R)-4-fluoro-3-methyl-1-oxo-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoindolin-2-yl)-5-oxopentanoate (synthesized by the method set forth in Example 2 starting with 4-bromo-3-fluoro-2-methylbenzoic acid). The crude product was purified by preparative LCMS (Column: Waters XBridge C18, 150 mm x 19 mm, particle size 5 μm; Mobile phase A: 5:95 acetonitrile:water (with 0.1% trifluoroacetic acid); Mobile phase B: 95:5 acetonitrile:water (with 0.1% trifluoroacetic acid); Gradient: 10% B hold for 0 min, 10-30% B over 20 min, then 100% B hold for 5 min; Flow rate: 20 mL / min. LCMS (Method B): Retention time 1.01 min, [M+H] + 369.1; 1 H NMR (400 MHz, DMSO-d6) δ 7.95 (t, J = 7.1 Hz, 1H), 7.81-7.62 (m, 2H), 7.22 (s, 1H), 7.09 (s, 1H), 7.04 (br d, J = 7.0 Hz, 1H), 6.97 (s, 1H), 6.78-6.72 (m, 1H), 4.93 (q, J = 6.5 Hz, 1H), 4.82 (dd, J = 12.4, 5.3 Hz, 1H), 2.91-2.83 (m, 1H), 2.75-2.60 (m, 2H), 2.36-2.32 (m, 1H), 1.52 (d, J = 6.8 Hz, 3H).

[0293] Example 28 (R)-3-((R)-5-(6-amino-4,5-dimethylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione [ka] To a stirred solution of Example 17 (250 mg, 0.531 mmol) in acetonitrile (4 mL) was added TFA (1 mL, 13.81 mmol). The reaction mixture was heated at 90° C. for 2 h. The reaction mixture was concentrated under reduced pressure and the crude product was purified by preparative SFC (Chiral Pak IC (250 x 50) mm, 5 μm; % CO2: 50%; % co-solvent: 50% of 5 mM ammonium acetate / ACN:MEOH (50:50); flow rate: 300.0 g / min; temperature: 40°C; UV: 240 nm) and the first isomeric fraction eluting with a retention time of 6.9 min was concentrated to dryness and lyophilized to give (R)-3-((R)-5-(6-amino-4,5-dimethylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (15 mg, 7% yield). LCMS (Method D): Retention time 1.28 min, [M+H] + 397.1; 1 H NMR (400 MHz, DMSO-d6) δ 10.97-10.89 (m, 1H), 7.99 (br d, J = 6.8 Hz, 1H), 7.57 (d, J = 8.0 Hz, 1H), 6.91 (s, 1H), 5.80 (s, 2H), 5.00 (q, J = 6.7 Hz, 1H), 4.76-4.67 (m, 1H), 2.82-2.70 (m, 1H), 2.67-2.55 (m, 2H), 2.22 (s, 3H), 2.03 (s, 4H), 1.53 (d, J = 6.8 Hz, 3H).

[0294] Example 29 3-((R)-5-(6-amino-5-fluoro-4-methylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione [ka] Example 29 was synthesized using general method 4 from 6-chloro-3-fluoro-4-methylpyridin-2-amine and tert-butyl (S)-5-amino-4-((R)-4-fluoro-3-methyl-1-oxo-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoindolin-2-yl)-5-oxopentanoate (synthesized by the method set forth in Example 2 starting with 4-bromo-3-fluoro-2-methylbenzoic acid). The crude product was purified by preparative LCMS (Column: Waters XBridge C18, 150 mm x 19 mm, particle size 5 μm; Mobile phase A: 5:95 acetonitrile:water (with 0.1% trifluoroacetic acid); Mobile phase B: 95:5 acetonitrile:water (with 0.1% trifluoroacetic acid); Gradient: 10% B hold for 0 min, 10-30% B over 20 min, then 100% B hold for 5 min; Flow rate: 20 mL / min. LCMS (Method B): Retention time 1.18 min, [M+H] + 401.3; 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 7.95 (t, J = 7.1 Hz, 1H), 7.60 (d, J = 7.8 Hz, 1H), 6.93 (d, J = 3.3 Hz, 1H), 4.90 (q, J = 6.6 Hz, 1H), 4.80 (dd, J = 4.9, 12.9 Hz, 1H), 2.89-2.78 (m, 1H), 2.70-2.60 (m, 2H), 2.24 (s, 3H), 2.07-1.97 (m, 1H), 1.51 (d, J = 6.5 Hz, 3H).

[0295] Example 30 3-((S)-5-(6-amino-5-fluoro-4-methylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione [ka] Preparation of 30A: 3-fluoro-4-methyl-6-(trimethylstannyl)pyridin-2-amine A stirred solution of 6-bromo-3-fluoro-4-methylpyridin-2-amine (0.05 g, 0.24 mmol) and hexamethylditin (0.076 mL, 0.366 mmol) in toluene (3 mL) was purged with argon for 5 minutes before adding [1,1'-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) (0.016 g, 0.024 mmol). The reaction mixture was stirred at 100° C. for 15 hours, cooled to room temperature, and filtered. The filtrate was concentrated under reduced pressure to give 3-fluoro-4-methyl-6-(trimethylstannyl)pyridin-2-amine (69 mg, 82% yield). LCMS (Method A): Retention time 1.68, [M+H] + 289.2.

[0296] Example 30: Stille coupling and cyclization was achieved following general method 2 using intermediate 30A and intermediate 16D. LCMS (Method D): Retention time 1.17 min, [M+H] + 401.3; 1 H NMR (400 MHz, DMSO-d6) δ 10.94 (s, 1H), 7.94 (t, J = 7.3 Hz, 1H), 7.58 (d, J = 8.0 Hz, 1H), 6.93 (d, J = 3.3 Hz, 1H), 6.31 (br s, 2H), 5.01 (q, J = 6.6 Hz, 1H), 4.72 (dd, J = 4.9, 11.9 Hz, 1H), 2.79-2.67 (m, 1H), 2.64-2.55 (m, 2H), 2.24 (s, 3H), 2.07-1.98 (m, 1H), 1.53 (d, J =6.8 Hz, 3H).

[0297] Biological Assays The pharmacological properties of the compounds of the invention may be confirmed by a number of biological assays. The following biological assays have been carried out with the compounds of the invention.

[0298] Helios cytolysis assay Jurkat cells were seeded at 80,000 cells / well in RPMI (40 μL) + 10% FBS in 384-well cell culture plates, to which test compounds had been added by ultrasonic acoustic dispensing technology. The cells were incubated and cultured for 72 hours at 37°C and 5% CO2. To facilitate the analysis, the cultured cells were spun down at 200 rpm for 5 minutes and the supernatant was removed. After shaking the plate to detach the cell pellet, the cells were resuspended in fixation buffer (50 μL, eBioScience FoxP3 buffer set, 00-5523-00) for 60 minutes at room temperature. After centrifugation and removal of the supernatant, the cells were permeabilized in permeabilization buffer (50 μL, eBioScience FoxP3 buffer set, 00-5523-00) for 10 minutes at room temperature. After permeabilization, cells were spun down and the supernatant was replaced with fluorescently labeled antibodies (20 μL) against Helios, Ikaros and Aiolos or corresponding isotype controls / 1x permeabilization buffer (Ikaros-Alexa488 [Biolegend, Cat #368408, 1:50], Helios-PE [CST, Cat #29360, 1:50], Aiolos-Alexa647 [Biolegend, Cat #371106, 1:25]) and the staining reaction was incubated for 1 h at room temperature protected from light. Then, 1x permeabilization buffer (30 μL) was added, cells were centrifuged and the supernatant was removed. Stained cells were resuspended in flow cytometry staining buffer (25 μL, PBS + 0.2% BSA) and analyzed using an Intellicyt Ique Plus flow cytometer.

[0299] [Table 9]

Claims

1. Formula (I): 【Chemistry 1】 [In the formula, R 1 is -NH 2 or -NH(CH 3 ) and Each R 2 are independently F, Cl, -CN, C 1-4 Alkyl, -CH 2 F, -CHF 2 , -CF 3 , -OCH 3 or cyclopropyl; Each R 4 are independently F, Cl, -CH 3 , -CH 2 F, -CHF 2 , -CF 3 or -OCH 3 and R 6 is hydrogen, C 1-2 Alkyl or C 1-2 is fluoroalkyl; m is 0, 1, 2 or 3; and Wherein n is 0, 1, 2 or 3; However, R 6 is hydrogen, then m is 1, 2 or 3. or a salt thereof.

2. R 1 Ga-NH 2 2. The compound according to claim 1, wherein:

3. R 1 -NH(CH 3 2. The compound according to claim 1, or a salt thereof.

4. Each R 2 are independently F, -CN, -CH 3 Or -CF 3 2. The compound according to claim 1, wherein:

5. R 6 But, C 1-2 Alkyl, -CH 2 F, -CF 2 H, -CF 3 or -CH 2 CF 3 2. The compound according to claim 1, wherein:

6. R 6 But -CH 3 2. The compound according to claim 1, wherein:

7. R 6 But, C 1-2 Alkyl, -CH 2 F, -CF 2 H, -CF 3 or -CH 2 CF 3 and m is 0; or a salt thereof.

8. R 6 But -CH 3 and m is 0; or a salt thereof.

9. Each R 4 However, independently, F, -CH 3 , -CHF 2 Or -CF 3 2. The compound according to claim 1, wherein:

10. Each R 4 But, F, -CH 3 , -CHF 2 Or -CF 3 and R 6 is hydrogen; and m is 1 or 2; 2. The compound according to claim 1 or a salt thereof.

11. Each R 4 But F or -CH 3 and R 6 is hydrogen; and m is 1 or 2; 2. The compound according to claim 1 or a salt thereof.

12. Each R 4 But F or -CH 3 and R 6 is hydrogen; and m is 1, 2. The compound according to claim 1 or a salt thereof.

13. The compound is: 2-Amino-6-(2-(2,6-dioxopiperidin-3-yl)-4-methyl-1-oxoisoindolin-5-yl)-4-methylnicotinonitrile (1); 2-Amino-6-(2-(2,6-dioxopiperidin-3-yl)-6-methyl-1-oxoisoindolin-5-yl)-4-methylnicotinonitrile (2); 2-Amino-6-(2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxoisoindolin-5-yl)-4-methylnicotinonitrile (3); 3-(5-(6-amino-4-methylpyridin-2-yl)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (4); 3-(5-(6-amino-4-(trifluoroethyl)pyridin-2-yl)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (5); 3-(4-fluoro-5-(4-methyl-6-(methylamino)pyridin-2-yl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (6); 3-(5-(6-amino-4,5-dimethylpyridin-2-yl)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (7); 3-((S)-5-(6-amino-4,5-dimethylpyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (8); 3-((R)-5-(6-amino-4,5-dimethylpyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (9); 2-Amino-6-((3S)-2-(2,6-dioxopiperidin-3-yl)-3-methyl-1-oxoisoindolin-5-yl)-4-methylnicotinonitrile (10); 2-Amino-6-((3R)-2-(2,6-dioxopiperidin-3-yl)-3-methyl-1-oxoisoindolin-5-yl)-4-methylnicotinonitrile (11); 3-((S)-5-(4,5-dimethyl-6-(methylamino)pyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (12); 3-((R)-5-(4,5-dimethyl-6-(methylamino)pyridin-2-yl)-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (13); 6-((3S)-2-(2,6-dioxopiperidin-3-yl)-3-methyl-1-oxoisoindolin-5-yl)-4-methyl-2-(methylamino)nicotinonitrile (14); 6-((3R)-2-(2,6-dioxopiperidin-3-yl)-3-methyl-1-oxoisoindolin-5-yl)-4-methyl-2-(methylamino)nicotinonitrile (15); 3-((S)-5-(6-amino-4,5-dimethylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (16); 3-((R)-5-(6-amino-4,5-dimethylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (17); 3-(5-(6-amino-3-methylpyridin-2-yl)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (18); 3-(5-(6-amino-5-fluoro-4-methylpyridin-2-yl)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (19); 3-(5-(6-amino-3-fluoro-4-methylpyridin-2-yl)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (20); 3-(5-(6-amino-5-fluoro-4-methylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (21-22); 2-Amino-6-((3R)-2-(2,6-dioxopiperidin-3-yl)-4-fluoro-3-methyl-1-oxoisoindolin-5-yl)-4-methylnicotinonitrile (23); 3-((R)-5-(6-amino-4-methylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (24); 3-((R)-5-(6-amino-4-(trifluoromethyl)pyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (25); 3-((R)-5-(6-amino-3-methylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (26); 3-((R)-5-(6-aminopyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (27); (R)-3-((R)-5-(6-amino-4,5-dimethylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (28); 3-((R)-5-(6-amino-5-fluoro-4-methylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (29); or 3-((S)-5-(6-amino-5-fluoro-4-methylpyridin-2-yl)-4-fluoro-3-methyl-1-oxoisoindolin-2-yl)piperidine-2,6-dione (30) 2. The compound according to claim 1, which is a compound of the formula:

14. The structure: 【Chemistry 2】 2. The compound of claim 1, or a salt, stereoisomer, tautomer or hydrate thereof, having the formula:

15. The structure: 【Chemistry 3】 2. The compound of claim 1, or a salt, stereoisomer, tautomer or hydrate thereof, having the formula:

16. The structure: 【Chemistry 4】 2. The compound of claim 1, or a salt, stereoisomer, tautomer or hydrate thereof, having the formula:

17. The structure: 【Chemistry 5】 2. The compound of claim 1, or a salt, stereoisomer, tautomer or hydrate thereof, having the formula:

18. The structure: 【Chemistry 6】 2. The compound of claim 1, or a salt, stereoisomer, tautomer or hydrate thereof, having the formula:

19. The structure: 【Chemistry 7】 2. The compound of claim 1, or a salt, stereoisomer, tautomer or hydrate thereof, having the formula:

20. 20. A pharmaceutical composition comprising a compound according to any one of claims 1 to 19, or a pharma- ceutically acceptable salt thereof; and a pharma- ceutically acceptable carrier.

21. A pharmaceutical composition comprising a compound according to any one of claims 1 to 19 or a pharma- ceutically acceptable salt thereof for the treatment of cancer.

22. 22. The pharmaceutical composition of claim 21, wherein the cancer is selected from colon cancer, gastric cancer, pancreatic cancer, breast cancer, prostate cancer, lung cancer, ovarian cancer, cervical cancer, renal cancer, head and neck cancer, lymphoma, leukemia and melanoma.

23. 20. An agent for reducing Helios protein levels, Helios activity levels, or Helios expression levels in a cell, comprising contacting a Helios protein with a compound according to any one of claims 1 to 19, or a pharma- ceutically acceptable salt thereof.

24. The method of claim 23, wherein the Helios protein is encoded by the amino acid sequence of SEQ ID NO: 1, 2, 3, 4, or 5.

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